Wt p $0 210 K E2lWorld Bank Discussion Papers AMarine Biotechnology and Developing Countrles Raymond A. Zilinskas Carl GustafLundin Recent World Bank Discussion Papers No. 151 Transport Development in Soutilern China. Clell G. Harral, editor, and Peter Cook and Edward Holland, principal contributors No. 152 Tle Urbani Environment and Popiulationi Relocation. Michael M. Cernea No. 153 Funding Mechanismsfor Highzer Education: Finanicingfor Stability, Efficiency, and Responisiveness. Douglas Albrecht and Adrian Zidermiai No. 154 Earnings, Occipationial Chioice, atid Mobility in Seymented Labor Markets of tIdia. Shahidur R. Khandker No. 155 Manag0ing External Debt in Dcveloping Counitries: Proceedings ofa Joint Seniinar,Jeddah, May 1990. Thomiias M. Klein, editor No. 156 Dev clopingr Agricultural Extension ior W`omen Farmers. Katrine A. Saito and Daphline Spurling No. 157 Aiakeningl the A'arke: ['iet N'am 's Econonoc: Transition. D. NI. Leipziger No. 158 I-l,7qe Policy duritng the Transition to a Mfarket Econoty: Poland 1990-91. Fabrizio Coricelli and Ana I(everiga, editors No. 159 International Trade anid the Environment. Patrick Low, editor No. 1 6i International Migration anld nterniational Trade. Sharoni Stanton lRussell and Michael S. Teitelbauni No. 1 61 Civil Service Re/orm atid the World Bank. Barbara Nunberg andJohlnl Nellis No. 162 Rmral Enterprics Dev'elopment in China, 1986-90. Antlhonv J. OdY No. 1 63 7'ie Balance bctu'ceni Plublii and Private Sector Activ'ities in the Dclivcry of Livestock Services. Din a L. Uminali, Gerslhon Feder, and Cornelis de Haan No. 1 64 How' Do N\'ational Policies A/hot Long-rui Grouthi?: A Rcscar.l A gcida. William Easterly, Robert King, P.oss Levinie, and Sergio IRebelo No. 1 65 Fisiheries Dc1 'lopncit, FisherieS Manjagement, arnd Externalities. Kichard S. johmnston No. I 6f) The Building Blo-ks ofPanrticipation. Testiny Botrom -u p Plamnm n. Micliael NI. Cernea No. 1 67 Seed Sy)stem Deivelopmnent: 7Iic Appropriate Roles of/ti'e Priilatc t,iid Public Settors. Steven Jaffec andJiteiidra Srivastava No. 1 68 Eniironimcntital XIanayientint antd Urbajn 1ildnerability. Alcira Krciniler anid Mohani Muniasiiigle, editors No. 1 69 Comnmnon Property Resoirces: .A Ahissing Dimension 0/ Dcv clopienwt Strate ics. N. S. Jodh a No. 170 .4 .Chminese Province as a Rfiionn Etpenvient: The Casc oflf-ainani. Paul Ni. Cadario, Kazuko Ogawa, and Yin-KaliTi Wen No. 171 Isulics tOr Ilnfrastriuctiurc Alaaemcnlei in tic 1990s. Arturo Israel No. 1 72 Japanese .National Railu'ays Prii'atizatioti Study. Koiclhiro Fukui No. 173 Tle Livestock Sector iti Eastcrn iiurope: Cuonstraints and Optportimiiics. Corn elis dc Haan , Tj art Schillhorin Van Veeri, arid KarenL Brooks No. 174 Asscssing Dvcielopment Finanic hisim4tions: A Puliic lInterct Analy si.Jacob Yaron No. 175 Rt'source Management and Pastoral institiution Budilding in ihe Wl'est Ajrican Sahel. Nadarajah Shanintigaratnani, Troiid Vedeid, Anine Mossige, arid Mette Boviii No. 1 70 Public anjtI Priva t Sectfor Roles iin Agricultural Researcih: Theory arid Experience. Dinia L. Uminali No. 1 77 Thie Reglatotry Impedinents tto tie Private Indiustrial Settor Development in Asia: A Comparative Stiudy. Deena Khatkhate No. 1 78 China: ReJrng Inetertolietal Fiscal Relations. Ranigo.pal Agarwala No. 179 Nilppon Telegraph arnd Telephtone Prittatizationi Study: Experience ol'Japart aid Lessonsl/or Deielopintg Countries. Yoshiro Takanio (Contitinued on the inside hack cover.) 2 1 0 = World Bank Discussion Papers Marine Biotechnology and Developing Countries Raymond A. Zilinskas Carl Gustaf Lundin The World Bank Washington, D.C. Copyright C 1993 The International Bank for Reconstruction anid Development/THE WORLD 1BANK 1818 H Street, N.W. Washington, D.C. 20433, U.S.A. All rights reserved Manufactured in the United States otfAmerica First printinig August 1993 Discussion Papers present results of country analysis or research that is circulated to encourage discussion and comlmuleint withinl the developmenit conununity. To present these results with the least possible delay, the typescript of this paper has not been prepared in accordance with the procedures appropriate to formal printed texts, and the World Bantk accepts no responsibility for errors. The findinigs. interpretations, and conclusions expressed in this paper are entirely those of the author(s) and shotuld nlot be attributed in alny manner to the World Bank, to its affiliated organizations, or to members of its Board of Executive LDirectors or the couLtries they represent. The World Bank does not guarantee the accuracy of tle data included in this publication and accepts no responsibility whatsoever for any consequence oftheir use. Aly maips that accompany the text have been prepared solely for the convenience of readers: the designations and presentationi of material in them do not imply the expression of any opinion vhatsoever on1 the part of the World Bank, its affiliates, or its Board or member countries concerning the legal status of any countrv, territory, city, or area or of the authorities thereof or concerning the delimnitation of its boundaries or its nationial affiliation. The material in this publication is copyrighted. Requests for permission to reproduce portions of it should be sent to the Office of the Pulblisher at the address shown in the copyright notice above. The World Bank encourages dissemination of its work and will norrnally give pemission promptly and, when the reproduction is for iioncoiiunmercial purposes, without asking a fee. Permission to copy portions for classrooml use is granted through the Copyright Clearance Center, 27 Congress Street, Salem, Massachusetts 01970, U.S.A. The complete backlist ofppublications from the World Bank is shown in the annual Index of Pnblications, which containis an alphabetical title list (with full ordering information) and indexes of subjects, authors, and countries and regions. The latest edition is available free of charge from the Distribution Unit, Office of the Publisher. The World Banik, 1818 H Street, N.W., Washington, D.C. 20433, U.S.A., or from Publications, The World Bank, 66, avellue d'lena, 75116 Paris, France. ISSN: 0259-2 1 iX Raymond A. Zilinskas is a consultant and Carl Gustaf Lundin an environmental specialist in the Land, Water and Natural Habitats Division of the World Bank's Environment Department. Library of Congress Cataloging-in-Publication Data Zilinskas, Rkaymonid A. Marine biotechnology and developinig countries / Raymond A. Zilinskas, Carl G. Lundin. p. cni. -- (World Bank discussion papers; 210) Incltudes bibliographical referenices. ISBN U-X2]3-2590-6 1. Marine biotechnology-Developing countries. 1. Lundin, Carl G., 1964- . 11. Title. IIl. Series. TP248.27.M37Z55 1993 333.95'2'(0)91724-dc2U 93-26137 CIP Foreword Nobel Laureate Abdus Salam has named this report, capability-building in biotechnolo- the twenty-first century, "The Biological gy is a complex, expensive and long-term Age." What he is alluding to, I believe, is effort that requires sustained funding until that the current evolution in biotechnology both the scientific base and the industry are will reach an even more exciting stage in the established. Governments must also promote next twenty years or so, profoundly affecting conditions supportive to the biotechnology the wellbeing of individuals and the econo- industry, such as cooperative R&D ventures mies of nations. Eventually, biotechnology between research facilities and the private will influence in some way virtually all of sector to transform research findings into mankind's productiveactivities-whetherthey commercial products or processes. This be agriculture, health delivery, environmental cooperation and support is crucial in biotech- remediation, industry or energy production. nology both (I) because of the inherent rela- It is important then to approach biotechnolo- tionship between discovery and application gy cautiously by planning wisely, acting (for example, when a particular genetic deliberately, and monitoring applications process is discovered, it introduces a process closely over the long term to ensure that of interest to industry at the same time); and these influences are constructive, benefitting (2) because of the need for continued scienti- both mankind and the environment. fic involvement in the industrial development "The Biological Age" can also be expected of that process (a situation quite unlike that of to benefit more than just the rich, industrial other technologies, regardless of how com- countries. Professor Salam and many others plex they are). have shown clearly that biotechnology is For example, a country can set up an particularly appropriate for the developing automobile industry by importing the exper- world. There are several reasons for this tise and technologies necessary to do so. favorable assessment: (1) many developing Setting up and operating the automobile countries are in the semi-tropical and tropical operation, as complex as it is, can be done climates that produce rich and diverse biolog- without involving research scientists in the ical resources, the substrate for biotechnology effort. Not so with biotechnology. Because and for sustainable economic development; the biotechnology industry involves the ma- (2) most developing countries have sufficient nipulation of living and therefore dynamic grounding in the biological sciences and biological systems to achieve products or technologies that underlie biotechnology to be processes, trained scientists that know these able to build capacity even in advanced tech- systems are necessary to solve the problems niques; and (3) entry costs for biotechnology inherent in operating them. The biotechnol- research and industrial application, although ogy industry, therefore, breaks down the significant, are lower than for other high usual delineations between basic research, technologies. applied research, development and manufac- In order to transform the promise of turing; and governments' support for all of biotechnology into reality, however, a gov- these activities is a major factor in its suc- erinent must make a considerable commit- cess. ment to building the requisite science and As this report points out, however, inter- technology capability. As is made clear in national agencies can provide crucial assis- Iv FOREWORD tance to complement and ease the process. ogy, developing countries will concurrently For example, assistance might take the form build the expertise that can be deployed to of supporting special training for scientists or investigate and solve local environmental industrial managers in technology transfer, problems, and to collaborate in solving global biosafety and intellectual property issues. problems as well. To date, World Bank involvement in bio- Fundamental to the development of bio- technology is rather limited, and entering an technology, and to any cooperative ventures, emerging field at its beginning is a departure however, is that governments, scientists, from previous Bank practice. There are private sectors, and international development several reasons for proposing this initiative. organizations must first learn of the potential First, the marine area is relatively neglected and problems of this new scientific endeavor, in Bank investments-only a small fraction of especially as they relate to the home coun- that channeled to land-based efforts. Support- tries. The express intent of this report, there- ing marine biotechnology, therefore, would fore, is to publicize the potentials of marine be a step toward balancing these levels of biotechnology for economic development and effort. Second, many developing countries problem solving; identify some of the possi- are island countries or have long coastlines; ble risks inherent in this field (and how they yet, most of these have not developed their can be avoided or controlled); and clarify marine potential, except for fisheries and various investment options for international some aquaculture. Marine biotechnology development organizations. The Environment could be one of the means toward sustain- Department presents this paper to further able, environmentally sound, development of these purposes and to encourage discussion of ocean resources. The remediation of polluted how the Bank can best lend its support to this coastal waters and beaches also could be done emerging area of science and technology, to via marine biotechnology. Third, solving the benefit of sustainable development. global enviromnental problems will require participation from all countries, including the developing world. Without worldwide co- operation, such problems as the 'greenhouse" 7t effect, pollution of the high seas, destruction of coral reefs, loss of marine biodiversity in Mohamed T. El-Ashry general, and certain water-borne infectious Chief Environmental Adviser diseases will defy solution. In the process of to the President and building their capability in marine biotechnol- Director of the Environment Department Contents Acknowledgements vii Introduction by Dr. Rita R. Colwell ix Note from the authors xi Executive Summary xiii Chapter 1 The marine environment 1 Oceans and ocean space 1 Marine life 2 Marine environmental damage and destruction 2 Detecting and monitoring oceanic events 3 Chapter 2 A primer in biotechnology 6 Classical biotechnology 6 Advanced biotechnology 7 Biotechnology and developing countries 9 Safety and biotechnology 10 Biosafety and marine biotechnology 15 Chapter 3 Marine biotechnology and its sub-areas 24 Definition of marine biotechnology 24 Marine biotechnology research and applications 24 Chapter 4 Options in marine biotechnology for developing countries 55 Existing scientific and technical capabilities 55 Applications of marine biotechnology in the short and medium terms 57 Chapter 5 Building capability in marine biotechnology 66 Building R&D capability 66 Applying research 69 Industry and biotechnology 69 The role of government in building capability in biotechnology 70 Discussion 71 Chapter 6 The experience of major international agencies in marine biotechnology and related areas 73 Food and Agricultural Organization 73 Inter-American Development Bank and other Development Banks 74 Intergovernmental Oceanographic Commission 74 vd CONTENTS International Centre for Genetic Engineering and Biotechnology 75 International Maritime Organization 76 United Nations Conference on the Environment and Development 76 United Nations Development Programme 77 United Nations Educational, Scientific and Cultural Organization 77 United Nations Environment Programme 78 United Nations Industrial Development Organization 78 World Bank 80 Discussion 83 Chapter 7 Exploring World Bank options for investments in marine biotechnology 84 Science and technology lending 84 Support of environmental objectives 86 Support for private sector development 87 Chapter 8 Conclusion 90 Appendix A Marine biotechnology and related R&D institutions in developing countries 91 Appendix B Special equipment requirements for advanced biotechnology 100 Appendix C Definitions of marine biotechnology by scientists in industrial and developing countries 102 Abbreviations, acronyms and data note 105 Glossary of technical terms 107 References 110 Acknowledgements This report would not have been possible donesia), Dr. Milagrosa R. Martinez (Philip- without the assistance, cooperation and other pines), Dr. Enrique C. Mateo (Peru), Dr. support by a rather large number of persons M.L. Lizarraga-Partida (Mexico), Dr. Patri- who supplied information about their coun- cio Bernal Ponce (Chile), Dr. T.S.S. Rao (In- tries or the organizations they work for, re- dia), Mr. P.M. Satheesh Seshaiya (India), viewed drafts of the report, informed us of Dr. Thiam Peng Teo (Singapore), Dr. Luiz scientific and technical progress and clarified R. Trabulsi (Brazil), Dr. C.K. Tseng (China) their implications, and in general encouraged and Dr. Xun Xu (China). us to persist in our endeavor. More specifi- Several professionals from intergovern- cally, we thank the following persons for mental organizations reviewed an earlier draft their assistance as noted. of this report and provided many useful Information about their countries' activi- comments. Some also described pertinent ties in marine biotechnology or related areas activities sponsored by their organizations. In was provided by Dr. Martin Abraham (Ma- these regards, we thank Dr. D. Bartley laysia), Dr. Gideon Abu (Nigeria), Dr. (FAO), Dr. V. Campbell (UNIDO), Ms. J. Shanta Achuthankutty (India), Dr. Alonso Douek-Hykin (World Bank), Mr. S. Garcia Cardenas Aguilera (Chile), Dr. Kamaluddin (FAO), Mr. V. Kotchetkov (UNESCO), Mr. Ahmad (Bangladesh), Dr. M.S. Andhale (In- G. Kullenberg (IOC), Dr. T. Matsusato dia), Dr. E. Montenegro Arcila (Chile), Dr. (FAO), Mr. M. Schneider (UNEP), Dr. G. Angel A. Baron L. (Dominican Republic), Tzotzos (ICGEB/UNIDO) and Mr. K. Venk- Dr. Enrique Bertullo (Uruguay), Mr. N.B. ataraman (UNIDO). Bhosle (India), Cr. Vanderlel Perez Canhos The scientific/technical reviewers of draft (Brazil), Dr. Saipin Chaiyanan (Thailand), reports included Dr. R.R. Colwell, Dr. T. Mr. M. Chandrasekaran (India), Dr. Dong- Veach Long 11, Dr. Mark Ragan and Dr. Suck Chang (Republic of Korea), Dr. S.T. T.S.S. Rao. Chang (Hong Kong), Dr. Young-Meng Last, we appreciate very much the help Chiang (Taiwan), Dr. Ho Coy Choke (Ma- we received from World Bank professionals laysia), Mr. Pang Daode (China), Dr. Chen while this project was under way and the kind Dou (China), Dr. Ossama M. El-Tayeb indulgence they granted our requests for (Egypt), Dr. S.O. Emejuaiwe (Nigeria), Dr. assistance. Our gratitude is especially directed Esther Fernandez A. (Venezuela), Dr. J.L.R. to Dr. Eric Arrhenius, Dr. Maritta Koch- Pino Gavifio (Peru), Dr. Twee Hormchong Weser, Mr. Jan Post, Dr. E.W. Thulstrup, (Thailand), Dr. Wei-Shang Ji (China), Dr. and Dr. Colin Rees. The fine editing by Ms. Koyoaki Katoh (Indonesia), Dr. Mohammed Charlotte Maxey is also gratefully acknowl- Moazzam Khan (Pakistan), Dr. Aida Lackany edged. (Egypt), Mrs. Trisnaui Dyah Listyawati (In- Introduction This report provides an analysis of the described. Many of these chemicals have potential of industrial applications of marine proven to be of potential pharmaceutical biotechnology and is focused on developing value. However, I estimate that far less than countries of the world. It makes clear that 1 percent of potentially useful chemicals from biotechnology's potential for remediating eco- marine living sources have been screened so nomic and social problems of developing far. The chemical compounds that remain to countries is extraordinary. However, it is be discovered represent one of earth's great- important to realistically assess what can be est treasures. done in selected areas of biotechnology. In addition to marine pharmaceuticals, en- Biotechnology is not a panacea to cure all ills hancement of fisheries via aquaculture offers of mankind, especially problems arising from great value. Overharvesting of the world population increase, overutilization of natural oceans is recognized as a global threat. The resources, and underutilization of human ability to produce transgenic fish and shellfish talent. However, some areas of biotechnology in culture, which grow faster and to a larger have extraordinary potential to address some size with more efficient utilization of nutri- of these problems-and marine biotechnology ents, is of particular value to developing is one of the most attractive of those disci- countries, not only as a source of food, but plines. also as export products. This report analyses The oceans cover about 75 percent of the the industry and offers some clear recommen- earth's surface. Developing countries are pre- dations for enhancement of marine fisheries. dominantly riparian and, for the most part, Other areas of marine biotechnology that depend on the oceans for protein, in the form are of great potential include marine bio- of fish, shellfish and seaweed. A resource remediation; for example, use of marine that has not been fully tapped, which repre- microorganisms to mineralize and degrade sents potential income in addition to fisheries, toxic chemicals spilled into the marine envi- is marine pharmaceuticals. The diversity of ronment. Research has been done that shows the marine environment in tropical areas is that enhancement of biodegradation can be well recognized. It is wide-ranging and of achieved by adding nutrients, mainly phos- unusual extent. Many developing countries phates and nitrates, to spills in coastal areas are blessed with resources in the form of and along shore lines. While much research extraordinary biological diversity. needs to be done before reliable, safe and In recent years, relationships of marine efficacious bioremediation processes are bacteria with corals, sponges, invertebrate ready for large-scale use, it is certain that animals in the marine environment, as well as these techniques can greatly benefit those seaweeds and marine plants, have been inten- countries where environmental contamination sively studied. Results from this research has reached threatening proportions. It is not demonstrate complex chemical interactions yet clear how bioremediation can be deployed between these bacteria and their hosts includ- in operations to treat offshore or open ocean ing systems of elaborate signalling and terri- spills. torial marking. In studying the chemicals The potential of marine biotechnology in produced by marine organisms, a variety of areas other than those I have mentioned lies toxins and chemical compounds have been in exploitation of resources in an environ- x INTRODUCTION mentally protective way. The value of bio- approach to biotechnology be applied in the technology is that the genetic capability of marine environment, coupling bioremediation marine organisms can be harnessed through and applications of biotechnology with the the cloning of the genetic material into sys- production of fish and shellfish through tems that can be manipulated and magnified aquaculture. in the laboratory, without depleting the re- The information provided in this document source in the natural environment. lays the groundwork for an effective and effi- It has been stated that the oceans are cient design of a plan of action for devel- underutilized in their capacity for feeding the oping marine biotechnology in all countries populations of countries of the world, but of the world, but developing countries that classical approaches to technology (that is, aspire to improve living conditions for their fish farming and seaweed farming) are limit- populations are likely to be the main benefi- ed to maximum yield much as is land agricul- ciaries. ture. However, with the application of genet- ic engineering, the productivity of the coastal areas can be greatly improved. The forces at play are the increasing populations of near- shore regions along with increased demand Rita R. Colwell, D.Sc. for commercially valuable marine products. President Thus, the future requires that a coordinated Maryland Biotechnology Institute Note from the authors This report focusses on a rapidly emerging niques can be used to break down pollutants, science-based technology-marinebiotechnol- to alleviate environmental damage. ogy. The term 'marine biotechnology" differs The intent of this report is to introduce in meaning among scientists. We think of marine biotechnology to nonspecialists, clari- marine biotechnology as a collection of re- fy its relevance to developing countries, and search and developmental activities in the outline the role of the World Bank and of biological, chemical and environmental sci- other international agencies in helping to ad- ences that occur in or are related to the vance it. However, marine biotechnology marine environment. Only a few specialized cannot be considered in isolation; after all, it facilities, located mostly in the United States is that intersect where biotechnology meets and Japan are dedicated to research exclusive- and overlaps with the marine environment ly in marine biotechnology. However, many and its living resources. A brief review of more laboratories have researchers working relevant aspects of the oceans and the life in marine biotechnology-related areas. they support and an introduction to the wider "Emerging," for the purpose of this re- field of biotechnology are useful as a starting port, means that the technology is at a stage point. Thus, Chapter 1 sets the stage for the in its developmental cycle when practical marine part of marine biotechnology by applications engendered by the technology are briefly describing and discussing certain being identified and laboratory processes and aspects of the marine environment that are techniques are being moved into practice. pertinent to biotechnology. In Chapter 2 is Looking at the term from another perspec- found a primer on biotechnology, consisting tive, an emerging technology is one that the of a review of its history, a presentation of public and its representatives begin to recog- recent developments in this field, and an nize as having the potential to generate new explanation of its "classic" and "advanced" scientific knowledge and produce useful new components. Special attention is given to products and processes. Marine biotechnolo- advanced biotechnology, including its work- gy research has produced a few applications force and equipment requirements and the to date, but its potential economic effects are possible risks biotechnology research, testing substantial, to be realized in five or ten years. and products may pose to scientific workers Many island and riparian developing and society. Chapter 3 is the heart of this countries are fortunate in possessing territo- review; it defines marine biotechnology and ries of subtropical and tropical marine waters provides a description of major sub-areas that shelter a large diversity of estuarine and (suggestions for how marine biotechnology marine life. At the same time, large areas of should be defined are found in Appendix C). their marine and coastal environments suffer Our consideration of marine biotechnology from the detrimental effects of manmade continues in Chapter 4, where options for de- pollutants. The largest promises of marine veloping countries in this field delineated and biotechnology are thus in two areas. First, analyzed. Specifically, each of the nine sub- some of its techniques may be deployed for areas of marine biotechnology is assessed in the sustainable exploitation of natural resourc- terms of promise for developing countries in es under environmentally sound conditions; the short and medium term and the degree of and second, other marine biotechnology tech- difficulty inherent in capability building (we xl NO TE FROM THE A UTHORS list institutes in developing countries whose solving and for economic development. We work programs encompass marine biotechnol- conclude the substantive part of the report ogy or related areas in Appendix A). Meth- with a short essay on why it is important now ods for building capability in biotechnology for developing countries to commit to capa- and in marine biotechnology are explained in bility building in marine biotechnology and Chapter 5 (some specific requirements for ad- why intergovernmental organizations should vanced biotechnology R&D are listed in assist in this endeavor. Appendix B). Examples of projects in marine This report is intended mainly for World biotechnology and related areas supported by Bank professionals who in the future may major intergovernmental agencies, including formulate and administer projects in marine the World Bank, are presented in Chapter 6. biotechnology. Thus, the language is largely In Chapter 7 we elaborate explicit proposals nontechnical, and when technical terms are for how the World Bank can promote marine used they are defined. In addition, a glossary biotechnology capability building in develop- of technical terms is provided. Those who ing countries, including ensuring that results wish to delve more deeply into the subject from research are applied in national problem matter may consult the references. Executive Summary The tremendous diversity of life in tropi- Marine natural products chemistry cal and subtropical seas represents the world's most abundant, but least utilized, As part of their metabolism, many marine living resources. Island countries such as organisms secrete compounds that help them Haiti, Indonesia and the Philippines, as well survive and that incidentally have properties as countries favored with long cost lines, beneficial to mankind. Screening programs including Chile, China, India and Sudan, have discovered algae, corals, sponges and have barely drawn on their marine capital. tunicates that produce compounds showing Thus, some of the poorest countries oversee antibiotic, anti-tumor, anti-viral or anti-in- potentially the earth's richest assets. flammatory properties. As procedures are improved, marine organisms producing anti- Marine biotechnology applications parasitic, pesticidal, immune-enhancing, growth-promoting, and wound-healing chemi- Marine biotechnology, which is defined as cals will certainly be found. This field has "the application of scientific and engineering substantial possibilities for growth since principles to the processing of materials by fewer than 1 percent of marine species have marine biological agents to provide goods and been screened. services," has many applications of impor- Bioremediation tance to developing countries-particularly aquaculture, marine natural products chemis- Microorganisms can be used to break try, bioremediation, biofllm or bioadhesion, down pollutants and wastes in soil or water to cell culture, biosensors and terrestrial agricul- harmless or less toxic end products. The ture. microbes used in bioremediation have usually been recovered from natural sites but have Aquaculture had their natural capability for breaking down pollutants enhanced through research and Marine biotechnology may benefit aqua- development (R&D). Most bioremediating culture in two major ways. First, its research microorganisms do not survive after the techniques can enhance a cultured organism's substance they feed on has been destroyed. growth rate, procreation proficiency, disease Because under normal circumstances biore- resistance and ability to endure adverse envi- mediation causes less damage to the environ- ronmental conditions. The organism's ability ment than do the present chemical and steam to grow and survive in intensive aquaculture cleanup methods, it holds significant advan- will thus be improved, increasing yields. tages over conventional techniques for clean- Second, through biotechnology vaccines may ing polluted harbors and waterways, as well be developed against bacterial and viral as decontaminating estuaries, mangroves and diseases that commonly afflict marine organ- other sensitive coastal communities. isms. Vaccines will protect fish, shrimp and other aquaculture organisms from diseases Biofilm and bioadhesion that now periodically decimate stocks, caus- A variety of marine organisms will settle ing enormous economic damage in Asia and Latin America. on surfaces exposed i seawater, eventually xiv MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES forming a crust. Organisms enmeshed in the monoclonal antibody or a DNA (deoxyribo- crust produce acids, which corrode piers, nucleic acid) probe. These biological mole- derricks and other structures. Encrustation cules are extremely selective; a monoclonal also increases hull drag in ships, decreasing antibody will, for instance, bind itself to only speed and raising operating costs. At present, one antigen, which may be a virus, a compo- paints containing heavy metals are used to nent of a bacterial cell wall, or a specific coat surfaces, preventing organisms from chemical. Kits based on monoclonal antibod- settling. However, toxic paints pose health ies or DNA probes are being used to quickly hazards to workers and pollute seawater. detect and accurately identify the pathogenic Marine biotechnology research seeks to bacteria causing cholera, shigellosis and clarify the molecular basis of the settling and typhoid fever, as well as viruses causing, for adhesion process, and the findings may be example, hepatitis. Biosensors may also be used to develop clean methods for preventing used in public health for such purposes as encrustation by marine organisms. identifying and monitoring substances and pathogens in the oceans, enabling scientists to Cell culture establish cause and effect relationships be- tween a particular agent and health events. Algae and other marine plant cells may be cultured in flasks, where they grow and Terrestrial agriculture subdivide much like bacteria. Cultured cells can be used to generate whole plants or they Marine biotechnology researchers seek to can be directed to synthesize natural prod- transfer characteristics inherent to marine ucts. Contrary to the chemical synthesis of animals and plants to their terrestrial counter- compounds, cell culture production is energy- parts. For example, the winter flounder conserving and essentially nonpolluting. survives sub-zero temperatures that would kill Many terrestrial plant cell culture systems are most animals, including other fish. The already producing pharmaceuticals, food flounder anti-freeze gene has been synthe- additives and pesticides; marine cell culture sized and inserted into yeast and higher systems will certainly be developed in the plants. Tests are under way to find out if it next few years along similar lines, which will will protect from sudden freezes crops grown be capable of producing agarose and other at high altitudes or northern latitudes. Anoth- valuable marine natural products. er example involves the world's most salt- tolerant plant, a microalgal species inhabiting Biosensors the Dead Sea where the salt content is 29 percent. Researchers are implanting genes Sensors are devices that detect a specific that code for salt tolerance in crop plants. substance or organism, and in biosensors the Success will mean that farmers can grow detecting element is a special biological rice, oil seed plants and other crops in soils material such as a chemoreceptor or an im- irrigated by brackish or salt water. munological molecule. An example of a chemoreceptor is the crab's sensing anten- Biotechnology and developing countries nule, which continually monitors water for dissolved substances ranging in chemical While biotechnology undoubtedly holds complexity from simple salts to pheromones. enormous promise for developing countries, In the second kind of sensor, an immunologi- that promise cannot be transformed into real- cal molecule, the recognition element is a ity until capability is built in all of the sup- Executive Summary xv porting processes needed. In particular, few up contracts, making startup funds available, developing countries have the broad and in- and guiding the partners in the equitable depth R&D infrastructure to undertake wide- sharing of intellectual property rights. ranging biotechnology research because they Research and applications involving genet- lack the trained scientific personnel, well- ically engineered organisms may present risks equipped laboratories and dependable supplies to workers, populations or other life forms. of rare, labile biochemicals. Even fewer have Risk can be minimized by applying safe the industrial capability to develop research procedures as specified by appropriate regula- results and exploit them. In view of these tions. Guidelines for biotechnology research problems, international agencies have a vital and field testing have been formulated by the role to play by providing technical assistance Organization for Economic Cooperation and and by catalyzing joint, cooperative R&D Development (OECD), and a UNEP/UNIDO/ efforts between laboratories and firms in WHO working group has elaborated guide- industrial and developing countries. lines for field testing applicable to developing Precedents exist in natural resource devel- countries. Either set of guidelines may be opment for fruitful cooperative ventures be- used as a model by governments when they tween R&D units in developing countries and formulate local laws. However, adapting counterparts or industries in industrial coun- foreign guidelines and regulations requires tries. Common to these projects is that both expertise in risk assessment and communica- sides benefit: the industrial country partner tion. Local scientists and regulators will have gains access to a raw material and retains cer- to be trained in these subjects, possibly in tain marketing rights, while the developing countries where regulatory frameworks have country partner gains expertise, financial been developed and the appropriate skills are backing and regional marketing rights. Some available. of these types of arrangements have been If wisely and correctly employed, marine brokered by international agencies, including biotechnology offers the tools for increasing the United Nations Development Programme high quality food supplies; for the sustain- (UNDP), United Nations Industrial Develop- able, environmentally sound exploitation of ment Organization (UNIDO) and World marine natural resources; for deploying Health Organization (WHO). These organiza- bioremediation to destroy or detoxify pollut- tions are particularly well placed to assist in ants harmful to people and the environment; developing profit-based options for joint and for improving public health through the ventures. Risks for failure or misunderstand- accurate detection and continuous monitoring ing can be minimized by the international of pathogens and pollutants in coastal waters. organization providing expertise in drawing 1 The marine environment This chapter has four sections. First, the Oceans can be considered as giant heat vertical and horizontal dimensions of oceans ponds, collecting and storing energy from the and ocean space are described. Second, the sun. However, because water is an efficient number, variety and uniqueness of marine life insulator, the heat collected from the sun is portrayed. Third, threats to the marine stays just beneath the marine microlayer, in environment and biodiversity are briefly a warm subsurface layer. Depending on discussed. Last, the technical means for de- geographic location, currents and other physi- tecting and monitoring marine events is cal factors, the subsurface layer can vary in recounted. depth, but will rarely exceed 20 meters. This layer supports a profusion of life, including Oceans and ocean space many species raised in aquaculture, and humans favor it for recreational activities. For this report, it is useful to view the The thermocline, the thin layer separat- oceans as having a horizontal and a vertical ing the surface layer from deeper waters, dimension. The horizontal has two aspects; marks a precipitous drop in temperature of 5° the high seas (blue water) and the coastal Celsius (C) or more. Beneath the thermo- margin. The latter is a complex system where cline, deeper waters support a profusion of land, sea, fresh water and atmosphere inter- life. However, as light is absorbed by water, phase. Most of the world's human population the biochemical and physiological characteris- lives in the earth's coastal areas, within what tics of many organisms inhabiting this layer is often termed the coastal zone. The coastal may differ from those found in surface wa- margin is thus heavily impacted by both ters. Many marine plants useful to aquacul- natural inputs and human activity. Under- ture will have their roots in deeper, cooler standably, coastal ocean waters are character- waters. ized by a high degree of variability in its Lowest in the water column is the cold biochemical and biological properties. water of the abyssal depth. The temperature The vertical dimension is the water col- of this water remains a steady 4°- 60 C umn. It is in effect a cross section of the throughout the world; it is rich in nutrients ocean space existing between the atmosphere and virtually free of pathogens. As is dis- and seabed. Oceanographers call the bound- cussed below, abyssal depths were once ary where ocean water meets with atmosphere thought to be lifeless, but recent explorations the marine microlayer. It is here where a have discovered a profusion of life forms, wide range of biological, chemical and physi- collectively called extremophiles, near the cal processes take place that vitally affect, for numerous hydrothermal vents that break example, nutrient distribution in ocean wa- through the seabed and release heat from the ters, the uptake of "greenhouse" gases from Earth's inner core (Grassle 1991). As dis- the atmosphere, and the gas exchange that cussed later, extremophiles have evolved provides oxygen to marine organisms. Oil re- unique mechanisms to survive that most leased in the oceans will drastically affect the inhospitable environment they populate, marine microlayer, principally through slick mechanisms that may prove extremely useful formation. to humanity. 2 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES Marine life the surface that depends on oxygen for ener- gy and carbon for building material, animals The oceans that cover 71 percent of the at these extreme marine enviromnents thrive earth's surface provide a haven for a multi- in the methane-sulfide and hydrogen-sulfide tude of biologically diverse life forms, rang- rich waters produced by the hydrothermal ing in size and complexity from the smallest vents. One interesting survival mechanism, known, the virus, to the very largest, the blue for example, is the unique symbiosis existing whale. Science has identified and character- between microorganisms and clams, mussels ized approximately 180,000 species of marine or tube worms. The bivalves pass the meth- algae, animals, bacteria, fungi and viruses; ane-rich water across their gills, where the and it is estimated that more than 800,000 are methane is picked up and processed by highly yet to be discovered. Unpolluted waters specialized bacteria that live only in the gills everywhere teem with life. It is hard to of this mollusc species. The free methane is imagine, for instance, the huge number of thereby made available to the host animal, krill that at favorable times occupy a given which uses it as its sole energy source. volume of the Antarctic Sea or the multitude The symbiotic relationship between the of squid that congregate off the coast of tube worm and bacteria is even more compli- California each spring. Nevertheless, the cated. The bacterium needs sulfur to survive, diversity of life found in tropical and subtrop- while the worm needs methane that the bacte- ical marine waters is greater than in colder rium processes from the seawater. The worm waters. These warm, richly endowed waters therefore has to pick up hydrogen sulfide fall mostly under the jurisdiction of develop- from the water and transport it to the bacteria ing countries. In fact, some of the least living within its body. Hydrogen sulfide developed countries may possess potentially would be toxic to the worm, except for the the most valuable marine resources. fact that it has evolved an unusual hemoglo- Although the basic building blocks of life bin that binds the chemical and allows the are the same, the conditions under which worm's blood to carry it safely to the bacteri- marine organisms live and propagate have um (Alper 1990). influenced their evolution, in the process endowing them with biochemical, biophysio- Marine environmental damage logical and genetic characteristics that are and destruction quite different from those exhibited by terres- trial life. Information about these characteris- The coastal waters of many developing tics, and the organisms that exhibit them, is countries are a haven for a large variety of mostly lacking. For example, scientists esti- marine life that provide aquaculturists, fisher- mate that less than 20 percent of terrestrial men and sailors with opportunities to earn a organism have been closely investigated; the livelihood, the sports-fishermen with recre- figure for marine organisms is considerably ation possibilities, and inhabitants in general less than 5 percent. with an important part of their dietary re- Life forms found in the abyssal depths quirements. However, over the last twenty or exhibit especially interesting characteristics. thirty years this resource has been severely Organisms living near submarine hydrother- misused and desecrated. Formerly profitable mal vents have evolved elaborate mechanisms fisheries for cod, salmon and sardines have to enable them to flourish in an extreme gone bankrupt due to overfishing; mangroves environment characterized by high pressure, in Asia have been destroyed to make way for high heat and no light. Unlike animal life on prawn farming; coastal waters off Africa and The mardne envIronment 3 Asia have been polluted by human and densities. If scientists are able to collect many slaughterhouse wastes, making them unfit for samples throughout a wide area and over a recreation, attracting sharks and otherwise longer period of time they may be able to creating public health hazards; riverain estu- gain an understanding of the distribution aries in Asia and South America, which in patterns of pelagic animals, the nature of the past provided breeding and spawning interactions between different species, and the grounds for an untold number of life forms, social structure of studied animals. Physical have been damaged or killed by agricultural samples, such as water and bottom material, and industrial pollution; and coral reefs can also be collected on-site for analysis of throughout the tropical seas, which protect constituent elements and pollutants. The seashores while furnishing shelter for an limitations of direct collection are: it is labor enormous variety of animals and plants, have intensive; it requires expensive equipment been dynamited or poisoned. and sophisticated instrumentation, including Damage to marine biodiversity caused by computers, for thorough analysis; the samples manmade activities ranges in severity from will always be minute compared to the area barely discernible to massive (UNEP 1990). or volume being sampled, so that making Marine tracts manifesting the heaviest dam- general hypotheses for the larger area is age are usually located near large population prone to error; and many important marine and industrial centers or close to the mining areas cannot be adequately sampled because or development of a natural resource, such as of political, physical or geographical factors. oil. Vast marine areas therefore cannot sus- Second, surveys can be done acoustically. tam traditional economic activities. Unfortu- Vessels equipped with acoustic systems can nately, unless governments institute conserva- detect schools of finfish and mollusks, includ- tion and anti-pollution measures and enforce ing krill, squid and midwater fish. Sophisti- them, the degradation of crucial ecosystems cated acoustic systems are often able to is certain to continue and the negative effects identify the detected animals. Acoustic sys- multiply. tems can be used to improve the efficiency of fisheries by locating and quantifying marine Detecting and monitoring living resources. However, sophisticated oceanic events acoustic systems are expensive and need trained personnel for their operation. These Oceanic events may be detected and moni- systems are not able to detect smaller organ- tored via on-site surveys and remote sensing. isms, nor organisms living on the bottom or Each set of technologies has its uses and at great depths. limitations. In the third method of on-site surveying, "sentinel" animals may be used to detect and On-site survey monitor pollutants, such as heavy metals, polluting chemicals and certain pathogens. On-site surveys are generally done in one These organisms can provide early warning of the following four ways. First, specimens of rising levels of pollutants or increasing of fish, plankton and other organisms may be number of pathogens, allowing public health directly collected through the use of nets or officials to institute preventive or corrective sampling chambers. Direct sampling allows measures before dangerous levels are scientists to identify which life forms popu- reached. For example, UNEP in cooperation late the site being sampled at a particular time with Intergovernmental Oceanographic Com- and to estimate population numbers and mission (IOC) in 1987 launched the Interna- 4 MARINEBIOTECHNOLOGYANDDEVELOPING COUNTRIES tional Mussel Watch experiment to use mus- tures; synthetic aperture radar for wave sels to detect and monitor chlorinated hydro- studies; microwave radiometers for wind carbons. The preparatory phase of the experi- speed; and coastal zone color scanners for ment was successful, so a full-scale mussel plankton densities. The remote sensing tech- sentinel program was begun in 1990 and will niques, when coordinated with surveys done continue through 1992 (UNEP 1991). There at ground level, allows scientists to determine are, however, some limitations to the sentinel and measure upwellings (the development of animal systems. In particular, sentinel ani- warm and cold water fronts), spot transition mals will detect only a small number of zones, and map areas of plankton blooms chemicals and pathogens, many sites will not (NRC 1985). Remote sensing would be a support the growth of sentinel animals, and rather expensive endeavor, if it included the often the detection of a chemical or pathogen cost of deploying sophisticated hardware in is post facto, after disaster has already struck. satellites that orbit around the world and The fourth method uses data acquisition installing receiving facilities with elaborate buoys that may be tethered at strategic sites equipment on the ground. However, most of to measure physical (wind speed, humidity, those are now in place and operate on a wave period and direction), chemical (con- communal basis. Remote sensing data and centration of salinity, dissolved oxygen and images can now be purchased from, for minerals) and biological (concentration of example, LANDSAT or SPOT, but the analy- nutrients such as phosphor and sulfur and sis of data requires special expertise. algal concentrations) parameters (Huglen While it is true that some of the tech- 1991). The sensors that detect and measure niques used, for example remote sensing, are parameters must interface with an operating sophisticated and products of high technolo- system that tells the sensors when to turn on gy, the data collected so far is elementary, and off and that processes data so it can be limiting possibilities for analysis of such transmitted to the operator, usually via satel- phenomena as climate change, the greenhouse lite transmission. effect and living resource depletion (see Figure 1). While more advanced remote Remote sensing sensing technologies are expected to come on line by the end of the 1990s (see Figure 2), Remote sensing can be done via satellites they also will have only limited ability to or aircraft; and in some instances, the equip- generate data on marine biodiversity, the ment and technique used for data collection productivity of marine organisms, the details and analysis are the same. Aircraft, though, of biogeochemical cycling, most of the pro- can in addition be used for detailed analogue cesses affecting environmnental change, the photography and visual interpretation for transfer of genes in the oceans, and many pursuit of fish and marine mammals (Baker other physical and biological occurrences. As 1991). Remote sensing equipment generates is discussed in the following chapters, ad- data on a wide segment of the electromagne- vanced biotechnology techniques may be used tic spectrum, which includes infrared radio- to overcome many of these shortcomings. meters for measuring the surface tempera- The nn enLvonment 5 Figure 1. Remote sensing and the environment: present technology SATELLITZES LANOSAT I-SS :Q; Q : _ im M f5 Q LW*wt TM SPOrPAN sparxs RAOAR(SAR ; 0 *;-00; ; NOAA AVHRR ''- ' I - '- NOAACZCS CO0---) HCmm -- 000-- AIRCRAFT AIRPHOTOS THERMAL SCANNERS ' Q ' O) '_ SAFVSLAR -0- 0 -el MSS SCANNERS ** ' - ' * * MAGIGRAVITY , SURFACE ION DETECTORS l e - - PFRS -: M S Excellent e Good 0 Fair - None A indirect only B cerlain conditions only ' non thernal Figure 2. Remote sensing and the environment: future technology YR SATELJrrES 91 ERS-1 ; ,,* -:O -, A 91 LANOSATO 91 SIR-ClX-SAR A Q 94 SPOT 3,4 P - * 94 SPOT 1 XS , -n *C) 91 JERS-t '- 0 I'0 * 92 MOS- 1 - U C7.1 95 AOEOS ' .* * * - * * * t 94 RAOARSAT , , ,., 90 KFA 1000 0'''-'' _9D COSMOSi 1J.* , - , ,* A 90 Eos - - s 96 SPACE STATION ,*,* *** *,- AIRCRAFr IMPFROVED MSS *; 9 MS RADAR - * MS THERMAL ; 0 Excellent 5 Good 0 Fair- None A indirect only Source: Baker 1991. EKNw53292SC 2 A primer in biotechnology Biotechnology may be defined as "the ings from Pasteur's research, industrialists application of scientific and engineering established a biotechnology industry in the principles to the processing of materials by early 1900s, which used fermentation to biological agents to provide goods and servic- manufacture on a large scale organic chemi- es." (Bull, Holt and Lilly 1982) Scientific and cals, such as acetone, butanol and ethanol. By engineering principles refer in the main to the 1925, 85 percent of all industrial solvents disciplines of microbiology, biochemistry, produced in the United States were manufac- genetics, biochemical and chemical engineer- tured via fermentation, a percentage that ing. Researchers in traditional applied fields dropped drastically after World War II be- such as agriculture, aquaculture and fisheries, cause they were replaced by synthetic pro- often employ biotechnology techniques to cesses based on cheap petroleum (Bjurstrom augment traditional procedures. While these 1985). During World War II chemical engi- are the essential characteristics of biotechnol- neers developed sophisticated fermentation ogy, a revolutionary advance in the field techniques to mass produce penicillin, fol- occurred in the early 1970s, when genetic lowed by other antibiotics. After the war, engineering was first developed. This, in further advances led to the manufacture of practical terms, marked a new departure for substances that were difficult to synthesize by the biosciences and bioengineering. Accord- chemical methods, such as steroids, enzymes ingly, biotechnology techniques developed and certain vitamins. before 1973 can be considered "classical' The research carried out by generations of techniques, while the many genetic engineer- geneticists, biochemists and other bio- ing techniques developed during the last scientists led to the development of the time- twenty years or so can reasonably be grouped tested techniques of mutation, selection and under the heading of 'advanced' biotechnolo- breeding that drive the conventional biotech- gy. For this reason, the primer begins with nology industry. Some of the recent industrial two sections addressing classical and ad- development programs were immensely vanced biotechnology. Next, the special successful; for example, the production promise that biotechnology offers for devel- capability of Penicillium notatum, the micro- oping countries is discussed. The chapter organism producing penicillin, has increased ends with two sections on biosafety in the over 5,000 percent since it was first used in terrestrial and marine environments. fermentation in the late 1930s. It is useful to describe a classical industri- Classical biotechnology al biotechnology project, one whose objective is to develop an antibiotic-producing organ- While humans have applied biotechnology ism. Such an organism may be discovered throughout history, at certain times "break- while screening many different microorgan- throughs" in research have led to remarkable isms recovered from, for instance, a soil advances in the field. The discovery by Louis sample from a tropical forest or a sponge Pasteur in 1857 that fermentation results from taken from a coral reef. After screening the action of microorganisms is a fine exam- uncovers an antibiotic-producing organism, ple of a breakthrough. Capitalizing on find- chemists assay its ability to produce the A primer In biotechnology 7 antibiotic. The natural (or wild) type organ- undesirable trait appears, a laborious ancillary ism does not usually release the antibiotic in breeding process called backcrossing is done sufficiently high concentration to be useful to eliminate it. for industry. Using low-producing organisms as parents, the industrial researcher develops Advanced biotechnology their progeny to produce greater quantities of the antibiotic than the parent. The first step is Advanced biotechnology is usually consid- usually to expose the parent organism to ered to have begun with the research on mutagenic chemicals, ultra-violet light or X- animal viruses undertaken by Paul Berg, rays. After treatment, microbiologists propa- Herbert Boyer, Stanley Cohen and associates gate potential mutants in culture and select at Stanford University and the University of those progeny that possess high antibiotic California, and reported in 1972 and 1973 producing ability for further selective cultur- (Cohen and others 1973; Jackson, Symons ing. Each selected strain is thereafter bred in and Berg 1972). In these experiments, recom- large numbers. Those strains that retain the binant DNA (rDNA) methods were devel- capacity to produce the antibiotic and that do oped, enabling the transfer of genetic materi- not exhibit other unwanted characteristics al, usually one or more genes, from one undergo further steps in the process. If devel- organism (the donor) to another (the host), opment is successful, chemical engineers where the genetic material became incorpo- propagate large numbers of the selected rated, or recombined, into the host's genome. progeny under controlled circumstances in Remarkably, the recombined gene retained industrial fermenters, where they produce the the same function in the new host as in the desired antibiotic in large quantities via donor. fermentation. The team that undertakes R&D The next significant advance in advanced projects in classical biotechnology will typi- biotechnology was in 1975 when the Argen- cally include microbiologists, biochemists, tinian scientist C. Milstein (at the time work- geneticists and fermentation engineers. ing in the United Kingdom) and the Swiss Scientists use classical biotechnology scientist G. Ko1hler discovered a method for techniques to alter the genetic makeup (geno- fusing two types of cells: the antibody pro- type) of microorganisms, plants and animals ducing B-cell from animals with a type of for the purpose of changing their physical cancer cell called myeloma (Kohler and characteristics (phenotype). Programs using Milstein 1975). The B-cells of an animal are classical techniques may be directed, for triggered by a foreign substance (called an example, to develop a faster race horse, a antigen) and begin producing antibodies. The higher-producing cereal plant species, or a antibody-producing B-cells are then fused superior antibiotic-secreting microorganism with the cancer cells. The fused cell, or strain. But because each operation in these hybridoma, produces a specific antibody programs affects the entire genome of the called monoclonal antibody against that target organism, many different genetic antigen. Antigens can be a variety of agents recombinations simultaneously occur; some or substances: virus, bacteria, a peptide, a could result in unwanted characteristics ap- sequence of DNA, a polysaccharide, and so pearing. The faster horse may, for example, on. The monoclonal antibody will attach to lack stamina; the higher-producing plant the antigen that stimulated its production. could be disease-prone; or the antibiotic- This property may be used by scientists to producing microorganism may prove unsuit- detect specific chemicals or parts of molecu- able for large-scale propagation. When an lar structures, by public health workers to 8 MARINE IOTECHNOL OGY AND DEVELOPNO COUNTRIES detect and monitor pathogenic microorgan- ple, strains of the bacterium Escherichia coil isms and toxic pollutants, and by industry to (E. colt), a microorganism normally found in capture and recover a specific substance from the gastrointestinal tracts of animals, have a solution. had human genes inserted in their genomes. Recombinant DNA and hybridoma con- Industry uses genetically engineered E. coli to struction were the first of a series of biotech- produce a range of proteins previously unique nology techniques that collectively are now to the human being, including human insulin termed genetic engineering. Genetic engineer- and growth hormone. These achievements ing has been defined by some as the science could not have been accomplished via classi- of manipulating genes and organisms to cal biotechnology. construct novel biological entities (Stone Second, genetic engineering can be spe- 1987). cific. Once the target organism's genetic Similar to classical biotechnology, ad- makeup has been characterized, bioscientists vanced biotechnology is in part driven by can target a specific gene or group of genes breeding and selection. Unlike classical for their attention. For example, research biotechnology, genetic engineering techniques may be directed at improving the yield of an may be employed to develop a novel biologi- amino acid-producing microorganism. Once cal entity (the transgenic organism); once the researchers identify the gene coding for the transgenic organism has been realized, scien- amino acid's sequence it can be defined, then tists employ breeding and selection to further cloned. Next, they insert multiple copies of enhance this organism's characteristics. As the gene in the original organism, or in before, chemical engineers use fermentation another well-characterized organism, perhaps processes for the large-scale production of the more suitable for industrial purposes. In wanted protein or metabolite. Scientists from either case increased production of the amino many different disciplines have contributed to acid will result. And to the point, only one the discovery and development of genetic aspect of the engineered organism's genome engineering, including biochemists, geneti- will have been manipulated- that bearing on cists, microbiologists, molecular biologists, amino acid production. physiologists and physicists. In addition, Third, and related to specificity, the fermentation and instrumentation engineers results from advanced biotechnology are apply research findings to industrial purposes. usually predictable. As noted above, classical While recognizing the long history of breeding often causes unwanted changes or biotechnology and the many benefits that traits because the natural recombination of classical biotechnology has generated for genes that occurs cannot be predetermined. humanity, advanced biotechnology equips Conversely, the genetic engineering recombi- bioscientists with new, powerful tools (UNDP nation in a well-characterized host takes place 1989). First, it offers the means for crossing according to plan and manipulation. The formerly impenetrable genetic barriers that problem of unwanted changes is minimized normally prevent crossbreeding between and will become more predictable as knowl- species. Thus genetic engineering can be used edge advances and techniques are perfected. to combine desirable characteristics of differ- Fourth, researchers using advanced bio- ent species. Industry now uses microorgan- technology techniques are able to achieve isms into which human or animal genes have results sooner than if they had used the classi- been spliced to produce several specialty cal approach of mutation, breeding and selec- pharmaceuticals that were barely known, or tion. For instance, the development of a high were unknown, a few years ago. For exam- yielding amino acid-producing bacterium A primer In blotechnology 9 through conventional breeding and selection Food industry may take several years; the same achievement could be reached within a year through the Genetic engineering is already making an application of advanced biotechnology tech- impact on the food industry. To illustrate, niques. formerly the only supply of rennin, an expen- sive substance vital to cheese making, was Biotechnology and the calf rumen. Now genetically engineered developing countries bacteria mass-produce it. Improved availabil- ity of rennin to cheese manufacturers means By now, about twenty years after the that they have more dependable supplies, are discovery of genetic engineering, modem able to improve the quality control of the biotechnology clearly holds immense promise final product, and can lower the cost of for developing countries, both to drive eco- production. nomic development and to provide tools for The possibility of building on existing solving problems related to health, food traditional methods used to produce ferment- shortage, and polluted marine and terrestrial ed foods and beverages also has relevance for environs (BOSTID 1982; UNIDO 1981). developing countries. For example, the food Four areas hold especial promise: agriculture, industry supports research on the genetics of food industry, pharmaceuticals, and industrial lactic acid fermenting bacteria to enrich the chemicals. flavor and nutritive content of fermented food staples. In another example, inedible biomass Agriculture may be used as a substrate for the production of single cell protein that, in turn, could be Animal husbandry will benefit from the used as a nutritive additive in livestock feed. introduction of recombinant vaccines and As is discussed below, marine biotechnol- diagnostic kits based on monoclonal antibod- ogy in general has much relevance to the ies and DNA probes. Other types of drugs food industry because marine organisms are useful to animal husbandry are growth hor- sources for many natural food additives, such mones produced by genetically engineered as preservatives, thickeners, supplements and organisms. Plant biotechnologists will im- emulsifiers. prove the growth characteristics of crops important to developing countries by endow- Pharmaceuticals ing them with improved disease resistance, improving their nutritional value, enhancing Industry produces about 80 percent of all their ability to withstand drought and heat or pharmaceuticals via chemical synthesis or by to grow in soils that have been damaged by extraction from animal and plant tissues. agrochemicals or that contain high concentra- Biological methods, usually fermentation, are tions of certain metals or minerals. Applica- used to produce the remaining 20 percent of tions from marine biotechnology, as will be prescription drugs. The proportion of fermen- discussed below, are likely to have uses in tation-produced drugs will increase dramati- terrestrial agriculture. For example, fish cally in the next ten to twenty years as the genes that code for anti-freezing proteins may new biotechnology techniques replace classi- be inserted in plants, allowing them to resist cal production methods. In addition, as men- frost and to retain their culinary properties tioned, unique pharmaceuticals produced by after freezing and thawing. genetically engineered organisms are creating 10 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES new niches in the drug market. To illustrate, going into developing microorganisms that at present approximately a dozen genetically destroy manmade toxic residues and sub- engineered drugs have reached the market stances (including petroleum and synthetic and are generating an estimated $2 billion chemicals) in soil and marine environments. income, but by 2000 the value of genetically engineered drugs is likely to exceed $10 While the promise of biotechnology billion. New drugs of particular importance should not be exaggerated and the difficulty to developing countries are: diagnostic kits of building capability is considerable, entry based on monoclonal antibodies and DNA into this field by developing countries is more probes (see page 48 and following) that en- readily accomplished than into any other high able doctors to rapidly detect and diagnosis technology field (Swaminathan 1991). For an infectious disease; and recombinant vac- that reason and also because most tropical cines that can prevent catastrophic diseases, and semitropical developing countries possess such as malaria, hepatitis, human and live- enormous natural resources amenable to stock trypanosomiasis and schistosomiasis. In development via biotechnology, their deci- fact, two recombinant hepatitis B vaccines are sionmakers would be remiss in failing to already in use. Others, including vaccines consider building capability in this field. against dengue fever and herpes, are likely to become available within five to ten years. Safety and biotechnology The benefits of a recombinant vaccine over traditional ones are that they are safer (be- After the introduction of rDNA technolo- cause they do not contain any parts of patho- gy, concerns arose about its safety. The genic materials), remain stable at ambient major worry was that an accidental or chance temperatures, and promise to be cheaper. recombination of genes would alter the bacte- rial host, endowing it with undesirable char- Industrial chemicals acteristics. The public, reflecting uncertainties by scientists, voiced their concerns about Approximately 90 percent of the substrates several aspects of biotechnology. Could an used to synthesize chemicals are petroleum- entirely new life form with unknown charac- based. For the many petroleum-importing teristics be created by researchers? Could countries the replacing of chemical synthesis otherwise innocuous bacteria accidentally by fermentation processes could have several become endowed with pathogenic properties beneficial effects. Fermentation occurs at during research and escape from research lower temperatures and pressures than chemi- laboratories? Could new recombinant forms cal reactions, thus saving energy and generat- of virus and bacteria cause pandemics of ing much less toxic pollution. Biotechnology unique diseases among man, animals or also can be adapted to use locally available plants? renewable natural resources, such as algae, agricultural and forestry wastes, and specific Laboratory safety crops to produce useful starting materials for many industrial chemicals, such as sugars, Most bioscientists believed that the possi- methane and alcohols. Fermentation processes bility of such events were small, but no hard adapted to use genetically engineered micro- data existed to buttress this conclusion. In organisms are producing single cell protein 1975, reflecting the depth of public concerns and certain amino acids, both valued as feed and considering the lack of information on additives in animal husbandry. Much work is the subject, scientists from throughout the A primer In blotechnology 11 world met in Asilomar, California (USA), to which has the initial responsibility for assess the risks of rDNA technology. The reviewing research proposals involving conclusions of the Asilomar conference were rDNA experiments presented by local used by the U.S. National Institutes of Health researchers and specifying the conditions (NIH) to formulate a set of guidelines. These under which these should take place. NIH guidelines for rDNA research were first Some difficult problems that require poli- published in 1976, but have since then been cy decisions are referred to the RAC. revised several times. Initially, the NIH guidelines: Although they were binding only on researchers funded by the NIH, the entire required total containment for rDNA United States scientific establishment quickly experiments and set forth the conditions accepted the NIH guidelines. Simultaneously, under which research could take place. the United Kingdom established the Genetic These ranged from the least secure condi- Manipulation Advisory Group (GMAG), tion, termed Biosafety Level 1 (BLI) to which formulated voluntary guidelines for high security containment, or BL4. Some British scientists. Soon, many other countries types of experiments were not allowed. adopted either the NIH or GMAG guidelines BLI and BL2 work require mostly com- for their own use or formulated national mon sense procedures, such as cleaning guidelines that were adapted from guidelines work surfaces, wearing laboratory existing elsewhere. smocks, and washing hands frequently. With the enactment of the NIH and These precautionary measures may be GMAG guidelines, scientists began to per- grouped under the rubric of good labora- form a series of risk assessment experiments tory practices that any well-run hospital to try deliberately to create pathogens. These clinical or research laboratory would confirmed that the possibility of accidentally routinely follow. Research considered creating pathogens in the laboratory was especially risky could only be carried out infinitesimal. They also proved that certain in BL4 facilities: self-contained units with laboratory procedures could be made safer entrance only through air-locks and with when genetic engineering was employed. all access rigidly controlled. All workers Specifically, when handling virulent viruses, in a BL4 laboratory must be specially scientists using genetic engineering techniques trained in the handling of extremely haz- could safely enclose viral particles in innocu- ardous infectious agents and must wear ous bacteria, something that could not be protective clothing (resembling space done with conventional means. For example, suits) when working. during hepatitis B research it is safer to enclose bits of the virus genome within an * created a national Recombinant DNA innocuous organism than to handle directly Advisory Committee (RAC), headquar- the whole virus as is done in conventional tered at the NIH, that reviews proposals research. for projects requiring the use of BL3 and Beyond directed risk assessment experi- BL4 facilities and adjust the NIH guide- ments, actual practice has demonstrated the lines in view of new scientific knowledge. adequacy of the NIH guidelines. Since the NIH guidelines first came into effect about * require each institution receiving govern- sixteen years ago thousands of research ment funds to set up and maintain an projects have been conducted throughout the Institutional Biosafety Committee (IBC), world in the agricultural, biological, indus- 12 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES trial, medical, microbiological and other and others. By now it is clear that the safety fields without apparent negative side effects. aspects of these products do not differ from This safety record indicates that genetic similar products produced by conventional engineering techniques are safe. means. In fact, biosafety regulatory programs There are three major reasons for the in the United States, the European Commun- safety of rDNA research. First, the successful ity and elsewhere assess inanimate products invasion, colonization and infection by a from advanced biotechnology on the same parasite that causes disease in the host is a basis as products from conventional research complex process. Not only is the number of and development. This is also the conclusion genes required to initiate infection large, but of WHO. For example, WHO would test a also the interactions between these many vaccine the same way, whether it was devel- genes are to a considerable extent dependent oped and produced using rDNA technology on their locations in a three-dimensional or a conventional cell culture system. No space. The probability of recreating this country to date has enacted new regulations complex milieu by accident when manip- aimed specifically at inanimate biotechnology ulating only one or a few genes is minute. products. Second, genes include regulatory DNA se- quences, called operons, that control in a Deliberate release positive or negative way the expression of the gene in each particular cell. It is difficult to The second concern, about deliberate imagine how these exceedingly specific release, has stimulated several studies to operons for disease-associated genes could be identify and quantify the risks associated with accidentally created. Third, the insertion of introducing genetically engineered organisms alien genes in a microorganism usually weak- into the environment. The two major risks of ens that organism in some way, diminishing deliberate release are: (1) direct harm to the its ability to compete with wild organisms or environment or any of its inhabitants, and (2) to survive the many stresses of natural condi- dispersal of the introduced organism and tions outside the laboratory. integration of its genes into the genomes of As data accumulated proving the safety of non-target organisms. biotechnology research, the RAC progressive- ly relaxed the NIH guidelines. Since the late NRC guidelines 1970s public fears about rDNA research in contained situations, such as laboratories, The U.S. National Research Council have largely disappeared. Other concerns, (NRC) has scrutinized the issues related to however, have surfaced about biotechnology the field testing of genetically engineered applications, specifically, the use of products microbes or plants in terrestrial situations and produced by genetically engineered organisms concluded that there are three essential crite- and the deliberate release of genetically ria for evaluating the risks associated with a engineered organisms into the environment. proposed release (NRC 1989): Product safety * Are we familiar with the properties of the organism and the environment into which Industry markets numerous products made it may be introduced? by genetically engineered organisms, includ- ing human insulin, human growth hormone, * Can we confine or control the organism various animal growth hormones, interferons effectively? A primer In biotechnology 13 What are the probable effects on the called Biotrack Information System. As of environment should the introduced organ- this writing, Biotrack has data on 650 field ism, or a genetic trait it carries, persist tests with no evidence of negative side ef- longer than intended or spread to non- fects.) target organisms? Case study: transgenic carp Specific methods for safely managing the field testing of genetically engineered organ- The three NRC criteria are central to the isms are in a state of evolution. In the United framework for risk evaluation in terrestrial States, proposals for testing genetically engi- situations and would apply equally to any neered organisms in the field are dealt with proposed release of genetically engineered on a case-by-case basis. In general, however, organisms into the marine or freshwater the process is as follows. For any proposal, enviromnents. It is illustrative to consider the a thorough environmental impact assessment one field test involving transgenic fish in light is conducted. The assessment addresses health of these criteria. To do this we need to re- and safety concerns by considering both view the events that preceded the decision to direct and indirect effects stemming from the allow the test and to scrutinize the-conditions proposed release. It must convincingly show under which this test is being performed. that the proposed release would probably not The fish in question is a carp containing a significantly alter or harm any aspect of the trout growth hormone gene. The transgenic environment or its biota. Permission for carp was developed by a team from the testing probably would not be granted if the Center of Marine Biotechnology (Maryland), organism to be tested was likely to present Stanford University (California) and Auburn high risk to non-target animals or plants; for University (Alabama) (Chen and Powers example, because it possessed characteristics 1990). At the end of 1989, the team request- such as enhanced fitness, increased pathoge- ed the U.S. Department of Agriculture nicity, or contained novel phenotypes. If the (USDA) to allow it to grow the transgenic project is judged to have a negligible impact carp in outdoor ponds to learn whether the on the environment, this finding is widely foreign gene affects the reproductive capacity publicized before a final decision is made in of the carp, whether the carp's offspring will order to give the public and its representa- inherit the foreign gene, and whether the tives an opportunity to scrutinize the environ- offspring will develop and behave as do the mental impact assessment report and to com- offspring from "normal" carp. This research ment on it. The regulating agency must take would be generally useful for improving the these comments, as well as statements made carp's genetic characteristics for aquaculture. by other interested parties, into account The field test proposal was strenuously op- before it makes its decision. At the time of posed by various environmental groups, this report, U.S. regulatory agencies have including the Foundation on Economic given final approval to over 400 field trials of Trends and the National Wildlife Federation, genetically engineered organisms, including a on grounds that carp have a significant poten- transgenic fish (see below). No negative tial to damage insects, plants and other fish in effects have so far been observed, indicating fresh water habitats. In view of the questions that the safety procedures seem to be work- raised, the initial proposal was remanded and ing, at least in the short term (Miller, Burris its drafters were asked to provide more and and Vidaver 1991). (Field tests throughout better information about possible environmen- the world are recorded in an OECD database tal impacts. 14 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES About six months later, the principal resembles testing in a closed system than true investigators submitted a redrafted proposal field testing. Nevertheless, it probably is a to the USDA. It asked for permission to raise model for the initial field testing of any 50,000 fry that had been spawned from nine aquatic animal or plant. As such, it is elabo- transgenic carp in ten outdoor pools. After rate and expensive, hardly an undertaking for three months, the number of fry would be most developing countries. (Safety consider- reduced to 300 per pond; these would be ations as they relate specifically to marine marked for identification and studied for the biotechnology are discussed at length in the next fifteen months. The fish would then be next section.) destroyed, before they reached sexual maturi- ty. The ponds stocking the fish would be International guidelines well-protected and would have no connection to any other waterways. During the last few years, safety aspects After some months of public hearings and of biotechnology has become a subject of deliberations, the USDA determined that the interest and concern by policymakers on the " .experiment with transgenic carp presents international level. The OECD has formulat- no significant risks to the environment" ed biotechnology guidelines to guide its (Transgenic fish 1990). It gave approval for member nations; guidelines appropriate for the experiment to proceed, and actual testing developing countries have been elaborated by began in June 1991. a working group established jointly by the For the purpose of this report, it is useful Food and Agricultural Organization (FAO), to review the conditions of the field testing of UNEP, UNIDO and WHO (UNIDO 1991). the transgenic carp in view of the three NRC These guidelines may be used by govern- criteria. First, in scientific terms, carp is ments as models for local laws. Although it probably the most studied and well-character- is too early to make a definite determination, ized of all fish species. The insertion of a the increasing involvement of international trout growth hormone gene would not change organizations in the biosafety issue may its physical properties, except that bearing on indicate a new trend; that is, governments growth. Whether the alien gene would change may be willing to manage biosafety through carp behavior is of course being tested. Since international cooperative efforts. (This ap- the testing is in effect being carried out in a proach makes sense since a genetically engi- closed, artificial system, the environment into neered microbe, plant or fish after release or which the transgenic carp is being introduced escape is not likely to respect national bound- is known. For these reasons, the first criteri- aries.) on is largely satisfied. Second, unless a While these efforts by international agen- deliberate, criminal attempt was made to cies are helpful, a difficult problem still faces release them, the conditions under which the governments of developing countries, because testing of the transgenic carp is taking place expertise in risk assessment and communica- precludes the possibility of their escaping. So tion is necessary in order for them to adapt certainly the second criterion is satisfied; the foreign guidelines and regulations to fit local tested organism is confined and controlled circumstances. However, these skills are effectively. The third criterion is not applica- typically scarce or lacking in developing ble since the test conditions preclude persis- countries. Local scientists and regulators tence or spread. should therefore be encouraged to acquire The field testing of the transgenic carp is them, but opportunities to do so are rare. so circumscribed and controlled it more Unlike the many training slots available to A primer In blotechnology 15 learn, for example, advanced biotechnology formulate policies for biotechnology research, techniques, risk assessment is not part of development and applications, they have most universities' curriculum. To some nothing to do with the safety of these activi- extent, this gap is being filled by a few U.N. ties. agencies. For example, the International Centre for Genetic Engineering and Biotech- Biosafety and marine biotechnology nology (ICGEB) and UNEP initiated a joint program on biosafety in 1991 (Practical As marine biotechnology develops and course 1991). Its two initial activities consist- advances, questions inevitably arise about its ed of courses offered in Trieste to researchers safety. In answer to this concern, explicit from developing countries: a three-day precedents cannot be found because of its course, "Genetically Modified Organisms: short history, but applicable lessons can be Safety in the Laboratory and the Environ- drawn from recent experience in the two ment" (July 1991) attended by thirty scien- fields that give rise to marine biotechnolo- tists; and a subsequent three-day course, gy-general biotechnology and certain marine "Genetically Modified Organisms for the applications. Accordingly, the next sections: 1990s," with fifty participants. Topics includ- (1) consider biosafety issues that biotechnolo- ed biological risk assessment, containment of gy in general has generated and analyze them genetically-modified organisms in the labora- in terms of their relevance to marine biotech- tory and the field, recommended procedures nology; (2) analyze special characteristics of for safe laboratory and industrial practices, the marine environment that bear on bio- transgenic animals, and analysis of existing safety; (3) scrutinize introductions of exotic biosafety legislation. The objective of these aquatic organisms into new environs; and (4) courses is to help scientists and regulators assess whether marine biotechnology poses from developing countries gain sufficient different safety and regulatory issues from skills in risk assessment and risk management those in terrestrial biotechnology. so they can return to their home countries and adapt these methodologies to fit local Biosafety issues in terrestrial biotechnology circumstances and conditions. It bears mentioning at this point that there In the preceding sections it was seen that is some concern that the biotechnology indus- biotechnology research raises one set of try will be a threat to producers of natural concerns, biotechnology products another. products. For example, in industrial countries Each requires reflection to gain a perspective cell culture systems that will mass produce on the possible biohazards they may generate. natural products, such as agar, saffron and vanilla, have been developed. The fear is that BIOTECH RESEARCH AND BIOSAFETY. As natural products will be displaced by biotech- discussed above, national guidelines that nology production systems, hurting the econ- regulate biotechnology research generally omies of the developing countries that pro- focus on containment and are voluntarily duce and export them. Similarly, European followed by scientists. The guidelines vary farmers have protested against the use of stringency of conditions under which research recombinant bovine somatotropin in animal may proceed depending on the level of risk husbandry, claiming that it would result in an believed inherent to the organism being overproduction of milk. While these are researched. When scientists work with a important socio-economic problems that need virulent pathogen they must do so in a high to be addressed by governments when they security laboratory and use elaborate proce- 16 MARINEBIOTECHNOLOGYANDDEVELOPING COUNTRIES dures to ensure the safety of themselves, environment pose a hazard to existing life other workers, and the surrounding communi- forms or the environment? ty. Conversely, research involving a non- Concerning the first question, most inani- pathogen usually requires no more than good mate products from biotechnology are known laboratory practices. It is probable that over chemicals or compounds produced via fer- 95 percent of all biotechnology research is mentation. However, rDNA technology has being done under the conditions defined by enabled the mass production of new products, good laboratory practices. such as human and other animal growth While marine and terrestrial organisms hormones, interferons and interleukens, may differ markedly in chemical and physio- which in turn have generated new marketing logical characteristics, the conditions under niches, changed the way we regard intellectu- which scientists conduct research are analo- al property and, at times, created ethical gous in marine biotechnology and terrestrial dilemmas. No biotechnology product is biotechnology. This is so because researchers known to have caused unique hazards. The in one field will be trained much like their main lesson from the experience gained by counterparts in the other; marine molecular governmental and intergovernmental agencies biology employs the same techniques as does when dealing with inanimate products, re- terrestrial molecular biology; and the labora- gardless of how exotic they are, is that the tories where R&D in both fields are per- testings of these products need not differ formed are similar, as is their equipment and from that of conventionally produced prod- reagents. Furthermore, having researchers ucts; the same criteria of safety and efficacy investigating a marine organism and a terres- apply equally to both. The strictness of the trial organism in the same lab does not in testing protocol will, of course, depend on itself create a special situation. The health the product's intended use. If the product is and safety issues posed by marine biotechnol- intended for animal or plant use, or is a ogy research performed in the closed system nonconsumable commodity, its testing would of a laboratory therefore can be expected to not be so rigorous. However, if the product be similar to those posed by comparable is a human drug, its testing would follow terrestrial biotechnology research. For these exacting procedures, including clinical phas- reasons, the voluntary guidelines that govern es. Summing up, so far governments have biotechnology research generally are also met concerns about the safety of inanimate expected to be pertinent to and adequate for biotechnology products by promulgating marine biotechnology research. To date, regulations that specify testing of the product scientists, public advocates and regulators and that establish mechanisms for monitoring seem to be in agreement on this point. the testing procedures. The chemical structures and other charac- BIOTECHNOLOGY PRODUCTS AND BIO- teristics of existing products from marine sAFETY. Advanced biotechnology research biotechnology are the same or similar to can produce two types of products-inanimate known compounds from the terrestrial envi- products and genetically altered living organ- ronment, although some have unique struc- isms. Each poses important questions, such tures. As more organisms from extreme as: Do inanimate products produced by genet- environments are collected, screened and ically engineered organisms pose risks (to investigated, uncommon compounds, showing humans, other animals or plants) above and antibiotic, anti-viral, anti-tumor and other beyond those posed by conventionally pro- properties, will be found. However, if the duced products? Would the deliberate release experience of terrestrial biotechnology is a of genetically engineered organisms into the guide, no matter how unique a product from A primer In biotechnology 17 marine biotechnology is, it is unlikely to graphic or geological barriers preventing the create a novel situation, or uncommon haz- spread of introduced organisms. Further, ard, that demands a new regulatory regime. except for the abyssal depths, ocean water is For example, if a unique marine toxin is never static; eddies, currents and wind are discovered, its physiological action is not forever creating movement. The continuity of likely to differ markedly from that of a oceans and the movement of water favor known toxin; neither will its toxicity be dispersal of organisms, whether by accident significantly greater than known toxins. or design (as noted in the previous section). Therefore, testing done according to estab- But an additional characteristic favors, if not lished procedures would unravel the chemical the dispersal of whole organisms, of genetic structure of the new compound, explain its material. This characteristic is that oceans are mode of action and, eventually, clarify its mostly water-salt water. Salt water is a effectiveness and safety. Similarly, when a medium that is kind to life, preserving the fish recombinant killed-vaccine is developed, viability of immersed organisms and, at its field testing would most likely follow times, parts of organisms by preventing established animal vaccine testing procedures; desiccation and deflecting deadly ultraviolet testing would be satisfactorily monitored by light. existing national regulatory authorities. Con- Organisms, and their parts, suspended in sequently, present protocols for testing prod- water and in ceaseless motion can easily ucts produced via conventional or advanced come into direct contact with other organisms biotechniques are appropriate for use in the and diverse suspended matter, creating possi- testing of marine biotechnology products. bilities for the dispersal of genes via one of These procedures can be improved on, per- three mechanisms: conjugation, transduction haps because they do not uncover all of the and transformation. properties inherent to a compound being tested, but this does not create special condi- CONJUGATION. For conjugation to take tions or hazards. place (that is, for two cells to directly interact The second question, the one that bears on to exchange genetic material), the cells have the deliberate release issue, is problematic. to be related. Thus, conjugations works No one has yet proposed the testing of a efficiently between two E. coli cells; fairly genetically modified organisms in the marine efficiently between two species in the family environment. Nevertheless, such a proposal Enterobacteriaceae, say E. coli and Salmo- could soon be submitted so it is not too early nella; but not at all or very inefficiently to consider the problems that it could gener- between, for example, bacteria and yeast. ate. To do so adequately, the special charac- The exotic bacterium could possibly thus pass teristics of the marine environment must first on the alien gene to a wild bacterium through be examined because they will determine how conjugation. Little is actually known about marine organisms and discrete genetic materi- conjugation among marine bacterial species, al (for example, plasmids and naked DNA) but it is reasonable to believe that dispersal disperse. by conjugation would be more likely in water populated by high numbers of bacteria due to Special characteristics of the marine contamination by sewage and human wastes environment that bear on biosafety than in blue ocean water. One of the most important characteristics TRANSDUCI-ION. The second mechanism is of oceans is that they, like the atmosphere, transduction, where a vector transfers the are continuous-there are therefore no geo- genetic material from one cell to another. For 18 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES example, the bacterial species Agrobacterium Biological invasion is "the arrival, estab- tumefaciens is a useful vector for transferring lishment, and subsequent diffusion of species genes into many plant species; simple virus- in a community in which they did not previ- es, called bacteriophages (or phage for short), ously exist in historical times. "(Carlton 1989) may transfer genes into bacteria (phages are Invasion usually results from range expan- specific, one type of phage will attack only a sion, which is the dispersal of the organism specific bacterial species). Results from by natural mechanisms. This phenomenon has recent research demonstrates that an immense not been studied extensively in the marine number of viruses and viral particles populate environment and, therefore, it is not yet the ocean surface layer (Proctor and Fuhrman understood. Since so little is known about the 1990; Suttle, Chan and Cottrell 1990). Mea- range expansion of wild species, no predic- surements show that one milliliter of surface tions can be made about the range expansion water contains between 107 and 109 viruses, of an organism, whether genetically engi- which means that the one millimeter thick neered or not, introduced into a site by hu- surface layer of the world's oceans would man activity. This lack of scientific data contain a total of 3.6 x 103 viruses (An creates problems for risk assessors; problems ocean of viruses 1990). The role of viruses in that cannot be resolved until much basic the marine environment is unknown, although research has been done to clarify this phe- it is believed that most of them are phages, nomenon. attacking species of marine bacteria, Introduction is the accidental or deliberate microalgae, plankton and other organisms. dispersal of organisms through human activi- ties (Carlton 1989). With the introduction of TRANSFORMATION. The third mechanism sailing, people have by chance or accident is transformation, where a plasmid or naked altered aqueous habitats throughout the DNA is taken up by a cell from the immedi- world's oceans, rivers and lakes. Ships have ate environment. Transformation usually carried organisms from one place to another occurs in the laboratory, where the researcher in their ballasts, encrusted on their hulls, and creates the chemical environment (media) bored in their wooden hulls. The openings of conducive to the reaction in a contained interoceanic and interlake canals have given vessel. Transformations appear to be excep- organisms added opportunities to migrate. tional phenomena in the atmospheric and Traders have carried crustaceans, fish and terrestrial environments; little is known about mollusks long distances from fishing grounds the dispersal of genes via transformation in to market places. Pathogens that afflict these the media that is ocean water. fishery products have been carried along (Carlton 1989). Marine species numbering in The dispersal of marine organisms the thousands have thus been moved across and their sequel the globe in innumerable patterns since trans- oceanic trading commenced. Past examples of dispersals of marine Besides accidental invasions, humans have organisms beyond their natural boundaries deliberatively introduced species to foreign provide us with information that can be locations as part of efforts to develop aqua- utilized to consider the dispersal of a geneti- culture and fisheries, as they have cultivated cally engineered marine organism, should one agriculture on land. Introductions have oc- escape in the course of field testing. There curred in waves throughout the 20th century are two types of dispersals; invasion and (Welcomme 1986), possibly in response to introduction. the changing tastes of consumers and as a A primer In biotechnology 19 result of the breeding of new, more desirable larva-eating fish may eat eggs and larva of fish, shellfish and crustacean strains. Thus, in other fish; the grass carp (Ctenopharyngodon the 1950s and 1960s there were large-scale idella) transmits a cestode-caused fish dis- deliberate introductions of fish and shellfish ease; the Pacific seaweed Sargassum muticum throughout developing countries. Introduc- was inadvertently introduced with C. gigas tions included the African 7ilapia to Asia and and eventually grew so dense along the Eng- Latin America; Indian major carps to South- lish and French coasts of the English Channel east Asia and Latin America; the mosquito that it interferes with other uses; and the larva-eating fish Gambusia affinis and Lebis- widely introduced shrimp Penaeus vannamei tes reticulatus through areas of the world in 1981 was found to be the carrier of the where malaria is endemic; and the black tiger pathogen infectious hypodermal and hemato- shrimp (Penaeus monodon) and the white poietic necrosis virus, which has decimated shrimp (Penaeus orientalis) throughout Asia shrimp aquaculture facilities throughout the and some Latin American countries. In the Pacific rim countries. late 1970s and early 1980s, large-scale intro- Sometimes an introduction appears suc- duction included striped bass (Morone saxa- cessful initially, but it proves the reverse over tilis) to the U.S. west coast; the Japanese the long term. Two examples of deliberate oyster (Crassostrea gigas) to the U.S. and introductions, undertaken with the best of Canadian west coasts and to France; Pacific intentions, that ended up disastrously have salmon (Oncorhynchus species) to Atlantic particular relevance to developing countries. waters; the pink salmon (0. gorbuscha) to One is the example of the introduction of the the Arctic Sea coast of Russia; a shrimp Nile perch Lates niloticus into Lake Victoria species from Panama (Penaeus stylirostris) to during the 1950s. The fish established itself Hawaii; and the Pacific seaweed (Undaria and in a few years local fishermen seemed to pinnatifida) to France (Sindermann 1986; benefit as they harvested 60,000 tons of the Welcomme 1986). More recently, in 1989, fish per year. But in the 1980s harvests the macroalgal species Euchema spinosum declined and scientists discovered that as have been introduced from the Philippines to Lates niloticus colonized Lake Victoria wa- Zanzibar, where it is used as a food condi- ters, it eliminated native cichlid fish stocks tioner. found nowhere else. In addition, the only Many of the deliberate introductions have practical way of preserving perch harvests benefitted local populations and improved the proved to be smoking, which demanded great economies of countries. For example, France quantities of wood, which in turn increased harvests over 100,000 tons of the Pacific the cutting of bushes and trees and led to oyster; the introduced fish Limnothrissa deforestation. By now it is clear that the yields about 4,000 tons from Lake Kivu and introduction of the Nile perch was destructive 12,000 tons from Lake Kariba; and Sri to aquatic and terrestrial biodiversity, while Lanka's entire inland aquaculture production the economic return from the introduction of 32,000 tons consists of introduced fish and could not be sustained. crustaceans (Sindermann 1986; Welcomme The second case is the introduction in 1986). The value of malaria larva-eating fish 1980 of the golden snail (Pomacea species) cannot be estimated, but it likely is immense. into the Philippines. The reason for introduc- On the debit side, like introduced species ing it was to provide farmers with an alterna- on land, aquatic introduced species have tive 'crop," the gourmet escargot, which caused damage ranging in severity from could be used locally for food and exported barely discernible to serious. The mosquito for cash. The export market never developed, 20 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES however, and local consumption is low. The * New diseases may be introduced along snail meanwhile thrives in the rice fields, with the deliberate introduction of a stock. where it turned into a pest, attacking newly transplanted rice plants and seed and destroy- * The introduced species may disrupt the ing up to 80 percent of the harvest. By the local aquatic community, leading to the end of 1991, 426,000 hectares of Philippine degradation of the immediate environment rice fields had been infested by the snail, (Welcomme 1986). which is resistant to pesticides and other control measures. The International Centre * Once an introduced organism colonizes a for Living Aquatic Resources Management locale, it may be impossible to eliminate. (ICLARM), headquartered in the Philippines, is trying to develop the integrated use of In view of the problems that dispersal of chemicals, biological control measures and marine organisms have engendered through- farming methods to control the snail (Lessons out the world, governments and international 1992). agencies have sought to prevent future prob- The foregoing indicates that the extent of lems and to alleviate existing ones through long-term damage from accidental or deliber- the adoption and implementation of codes and ate introductions of exotic animal species into rules. One of the most important of these is the marine environment often cannot be the Revised Code of Practice to Reduce Risks reliably assessed at the time of introduction; for Adverse Effects Arisingfrom Introductions nor is it possible to determine with certainty and Transfer of Marine Species adopted by whether the benefits stemming from deliber- the International Council for the Exploration ate introductions ultimately will outweigh of the Sea (ICES) in 1973 (and revised in costs. More specifically, any or all of the 1979). Other codes of practices, position following problems may result from introduc- statements and conventions on the subject tions: have been made by the American Fisheries Society (1973), the U.N. Conference on the * An introduced animal may disrupt local Law of the Sea (1982), the Council of Europe fauna through competition or predation. In (1984), FAO's European Inland Fisheries the worst case, the introduction of an exotic Advisory Commission (1984) and the Interna- species may lead to the extinction of the wild tional Union for Conservation of Nature and species. Natural Resources (1987). The overriding objective of these codes and statements is to * Genetic degradation of the host stock may direct concerted international action toward result from its introduction into a new locale. preventing accidental introductions and ad- Conversely, important genes may be lost if verse effects from deliberate introductions. the exotic species displaces or replaces the Due to the uneven implementation of the wild species. ICES code by nations, Dr. Carl Sindermann has suggested strategies for dealing with * An introduction may lead to a loss of future proposals for introductions. The over- population identity. In other words, when an riding strategy is for U.N. agencies and introduced species breeds with the wild nongovernmental organizations to educate the species inhabiting a locale, the adaptions for public, policymakers and national regulatory survival that the wild species have evolved agency personnel about the potential damage may become diluted or disappear in hybrid that the importation of a non-indigenous progeny. species can do to native stocks and the local A primer in blotechnology 21 environment. Countries will thus learn that it biotechnology. Present risk assessment and is in their best economic interest to have a management schemes, as well as existing strong regulatory regime to prevent unautho- regulations, seem to adequately cover these rized introductions and to delineate the condi- areas of marine biotechnology. tions under which authorized introductions The difference, then, is in field testing of may proceed. Another strategy is suggested genetically engineered organisms. As noted, for larger, industrial countries. It emphasizes USDA has given permission for the field regional approaches to controlling the transfer testing in enclosed ponds of only one geneti- of organisms, where the federal government cally engineered aquatic organism-a trans- ensures uniformity and continuity. genic carp; and because that testing is being Whatever approach is adopted, it should carried out in a closed system, it does not be implemented according to the general have much relevance to a future field testing operating principles set forth in the ICES in the marine environment. Here the tester code. These are based on the assumption that and the regulator faces special problems not risks from introductions are never zero. This faced by those performing field testing in being so, national regulatory regimes should closed chambers, because it may be impossi- be designed so as to minimize risks from ble to secure the biological isolation of the proposed introduction. organisms being tested. Biological isolation Risk reduction includes the thorough study cannot be guaranteed because of the three of the organism proposed for introduction in characteristics of the marine envi- its native habitat; consideration of developing ronment-the continuity of the oceans, the native stocks as an alternative to introducing perpetual motion of the water and suspended a new stock; and establishment of mecha- particles, and the potential for gene disper- nisms for monitoring the introduced stock sion via unfamiliar biological mechanisms. continuously. It is particularly important that Further, the lesson learned in the past from the scientific implications of a proposed dispersals of exotic organisms is that if and introduction be analyzed before the event, when the marine organisms being tested including clarifying ecological considerations escapes, the consequences are incalculable. (such as competition and predation); genetic Referring again to the field testing of the considerations (including potential for hybrid- transgenic carp, rather than consider the ization and change in gene frequency); behav- conditions of the field test itself, it is perhaps ioral consideration (including interactions more useful for our purposes to speculate on between the introduced and native species); the consequences of a successful test; that is, and pathological considerations (including the if the tests convincingly demonstrate that the possibility that the introduced species will standard culturing of the transgenic carp is carry along with it new infectious diseases) more cost-effective than culturing present (Sindermann 1986). stocks, what then? One conceivable scenario is that someone would try to take advantage Comparison of terrestrial and of the higher performing characteristics of the marine biotechnology transgenic fish by intensively culturing them in cages or pens emplaced in a lake or a Does marine biotechnology pose different river. The possibility of some transgenic fish safety and regulatory issues from terrestrial escaping would be high. What would be the biotechnology? As was discussed previously, consequences of an escape? marine biotechnology research does not, Referring to the six possible problems that neither does inanimate products from marine past introductions have caused, the conse- 22 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES quences can range in severity from no effect ture. Fish may be sterilized by two methods. or minimal to severe. If past experience of First, certain hormones can be administered terrestrial field testing of organisms that had to fish embryos, which render them sterile. a single gene inserted in their genome is a Researchers do not favor this method since it guide, no ill effects would result. However, cannot achieve 100 percent sterilization and we cannot completely discount the possibility hormonal residues may contaminate food- that the escape may trigger a low probability, fish. Second, fish eggs can be treated so the high consequence sequel, such as the follow- progeny are triploid; that is, each fish carries ing scenario: three sets of chromosomes rather than two (see page 28). Triploids are sterile. For ... the dangers of genetic manipulations added safety, triploid induction can be com- should be recognized, and biotechnolo- bined with further treatment that produces an gy may prove to be as much a threat to all female progeny. Triploid females are 100 natural species and genetic diversity as percent nonfertile. it is a justification for maintaining that Kapuscinski points out that even if only diversity. The release of individuals sterile transgenic fish are cultured, some risk with artificially composed genetic remains because of the necessity to maintain makeups into wild populations of the transgenic broodstock. The answer is to same species could upset the natural maintain broodstock in secure containment distribution of that species as well as facilities, and to educate everyone who works the competitive interactions with other with them of the ecological problems that species, destabilizing natural biological have resulted from introductions of exotic communities (Ihorne-Miller and Cat- fish species in the past. ena 1991). Of course, transgenic organisms other than transgenic fish may be the first candidate The field testing of a transgenic fish is not for field testing. In view of the research that likely to be proposed in the short term if for has already be done to genetically engineer no other reasons than technical ones. Accord- bacteria (for bioremediation) and microalgae ing to Dr. A. Kapuscinski, who works for the (for increased production of food additives), Department of Fisheries and Wildlife, Uni- they rather than multi-celled animals should versity of Minnesota, technical barriers must be considered as the primary candidates for be overcome and environmental risks reduced the first marine field testing. Further support before transgenic fish can be cultured (Kapus- for this contention comes from the report that cinski 1990). The technical barriers relate to the firm Envirogen Inc. in New Jersey is the cost-effective transfer of valuable genes preparing a proposal for the field testing of a and promoters into large numbers of fish; the bacterium that has been genetically engi- ready identification of transformed individuals neered to improve its ability to degrade the among the treated group; and the selective industrial pollutant trichlorethylene. Report- breeding of transformed fish to develop edly, the Envirogen proposal will be present- superior progeny. She estimates that it will ed to the EPA sometime during 1992 (First take a minimum of 10 years to overcome rDNA 1991). these technical barriers. The initial "field testing" of a genetically Once technical barriers have been over- engineered bioremediating bacterium would come, the major means by which risks related probably be done in a closed system, similar to transgenic fish may be reduced could be to to the one used for the testing of the trans- sterilize all fish slated for outgrowth in cul- genic carp. The strain to be tested may be A primer in biotechnology 23 "weakened" so it would not survive in the of dispersal of organisms and genes in the wild should it escape. Parameters that could marine environment and a satisfactory risk be tested in a closed system include surviv- assessment methodology for field testing in ability outside the laboratory, ability of the the oceans has been developed. Without organism to degrade trichlorethylene under doubt, for the present it is more difficult to various conditions and in the presence of evaluate and determine the possible effects of sundry chemicals, and whether synergism is the field testing in the marine environment of possible between the tested organisms and transgenic marine animals, plants and micro- other microorganisms. Such responsible organisms than in the terrestrial environment. testing would not likely endanger man or To conclude this chapter, the expectations environment. and problems of marine species field testing The field testing of a transgenic bacteri- today are approximately the same as it was um, and other microorganisms, should for the for terrestrial and/or atmospheric field testing present not be done in an open system. For when these activities were commencing some one, little is known about the dispersal of ten years ago. Unlike former times, however, whole microorganisms in the marine environ- we are at this time able to access the experi- ment, thus any or all of the six problems ence of past field tests and draw lessons for listed above could result. In addition, next to future marine field testing. Today's scientists nothing is known about the functioning and are thus better prepared to prepare compre- efficiency of the three mechanisms for gene hensive environmental impact statements dispersal in the marine environment, so no prior to testing based on risk assessment one could predict whether the alien genes methodologies adapted for the marine envi- carried by the transgenic microorganism ronment, design safe test protocols, and to would disperse, the frequency of possible institute efficient mechanisms for monitoring dispersal, the probability of dispersed genes test events and the long-term effects of tests. being taken up by wild organisms, or the Lessons from the past also strongly indicate ultimate effects of dispersal. For these rea- that it is in the vital interest of each country son, the field testing of organisms in the to have an effective, comprehensive regula- marine environment should be deferred until tory regime in place that will ensure proper research in biological oceanography, micro- precautions are taken, while preventing poor- bial ecology and environmental toxicology ly planned and executed field tests from being have clarified the details of the mechanisms undertaken. 3 Marine biotechnology and its sub-areas Although much R&D related to the marine often efficacious and desirable to merge environment has been, and is being, done traditional activities with new developments throughout the world, it is only recently that in biotechnology. a subset of these activities has been termed marine biotechnology. Marine biotechnology research and applications Definition of marine biotechnology Although thousands of projects have been In a strict sense, "...any scientific investi- or are being undertaken that can be consid- gation that focusses on marine organisms and ered as marine biotechnology R&D, it is that utilizes new cell, protein and nucleic acid possible and convenient to divide them into technologies such as recombinant DNA, nine sub-areas: aquaculture, marine animal hybridoma/monoclonal production, protein health, marine natural products, biofilm and engineering, polymerase chain reaction, and bioadhesion, bioremediation, cell culture, DNA hybridization" can be called marine biosensors, biological oceanography including biotechnology. However, this definition is public health, and terrestrial agriculture. believed too narrow by many researchers who hold that a host of different R&D activities Aquaculture rightfully are sub-areas of marine biotechnol- ogy. Taking into account this lack of consen- Aquaculture is a general term that refers sus on what marine biotechnology constitutes to the husbandry of aquatic (brackish-, fresh- and encompasses, we do not draw rigid and saltwater) animals and plants at densities disciplinary lines in this report but instead that are greater than those found under natu- view marine biotechnology as a field that ral conditions (FAO 1991). The development encompasses a patchwork of scientific and of the world's aquaculture sector has been technological activities relating directly to impressive, having increased production from marine organisms or their parts and that approximately 10 million tons in 1985 to 14 employ biotechnology techniques. Thus, a million tons today, and estimated at 22 mil- broad definition is analogous to the definition lion tons by 2000. If these projections hold of biotechnology given in Chapter 2 (Bull, true, FAO estimates that by the end of the Holt and Lilly 1982). Marine biotechnology century aquaculture products will account for then can be defined as 'the application of 20 to 25 percent of the world fisheries pro- scientific and engineering principles to the duction by weight, and in excess of 50 per- processing of materials by marine biological cent by value (FAO 1989). In comparison, agents to provide goods and services" (for the yield from world fisheries was approxi- other definitions, see Appendix C). mately 99.5 million tons in 1989 according to When biotechnology techniques are used FAO (1989 FAO statistics 1992), which is for research in certain applied fields, such as very near to its estimated maximum sustain- aquaculture, fisheries and natural marine able yield of about 100 million tons per year. products, they are included under the rubric Aquaculture has been practiced in some of marine biotechnology. This point is impor- societies for millennia, usually in ponds tant because, as will be discussed below, it is holding low-density populations of finfish, Marine biotechnology and fts sub-ares 25 shellfish or crustaceans (Costa-Pierce 1987). may pollute streams and lakes in which they More recent is the development of the so- are released. Once operations are under way, called 'intensive' aquaculture, which may be culture systems may become populated with defined as a condition when the density of contaminants that compete with crop plants fish exceeds 1 kilogram of fish per 57 cubic for food and sunlight (Gellenbeck and Chap- decimeters of water or 1400 kilograms of man 1983). Further, wastes from seafood shrimp per 0.4 hectares of water (McCoy processing plants associated with aquaculture 1990). For example, in culturing prawns in can create disposal problems; wastes usually the Philippines, the stocking rate for the giant are 20-25 percent for finfish and 80-85 tiger prawn, Penaeus monodon, in ponds percent for shellfish (Technical Q&A 1991b). where traditional techniques are used is about These problems do not relate directly to 10,000 prawns per hectare; in an intensive marine biotechnology and will not be dis- system the stocking rate is 100,000-300,000 cussed further. However, conditions created prawns per hectare (Primavera 1991). The by intensive aquaculture do create problems primary products of aquaculture are food for that may be solved or alleviated via the human consumption or natural products application of biotechnology techniques. useful as biomedical reagents, medicines, Thus, what will be discussed here is the food additives, and jewelry. The culture of deployment of marine biotechnology to in- ornamental fish is a relatively new but rapidly crease yields from aquaculture or to enhance growing area of aquaculture. the quality of its products. In particular, Mariculture is that subset of aquaculture biotechnology offers new methods to improve that is practiced in salt water. The term the heath status of cultured organisms (see mariculture is specific and will not be used in next section) and to regulate growth and this report unless there is a need to exclude reproduction of commercially important from the matter under consideration all but finfish, crustaceans, bivalves and algae. A marine organisms. discussion of specific marine biotechnology The practice of aquaculture consists of the applications to these organisms follow. application of a set of low-technology endeav- ors pertaining to the breeding, propagation, GROWrH AND REGULATION OF FINFISH. harvesting and marketing of algae, fish, One of the early successes of advanced bio- crustaceans, bivalves and gastropods. The technology was the manufacture of human major problems encountered by aquaculturists growth hormone (hGH) by genetically engi- relate to the siting of the culture ponds neered E. coli. Before this accomplishment, (Bailey 1988; Fernandez-Pato 1989; Prima- the only source of hGH was pituitary glands vera 1991). In particular, the prospective recovered from human cadavers. Not only aquaculturist needs to know the composition was the extraction of the hGH from the gland and chemical characteristics of the soil in difficult, but also approximately 5000 of the which ponds will be constructed, the quantity minute glands were required in order to and quality of the water in which animals will produce one gram of the substance. After grow, and the ready availability of animal techniques were perfected for the large-scale feed. Further, incorrectly sited ponds may manufacture of hGH produced by genetically severely damage the nearby environment. For engineered bacteria, or recombinant hGH for instance, pond construction has often entailed short, sufficient quantities have become the destruction of mangroves; sweet water available to treat all who need it and to fully used in hatcheries may deplete underground supply medical researchers. aquifers of fresh water; and untreated efflu- The developmental process that resulted in ents and wastes from farms and hatcheries recombinant hGH has been duplicated in 26 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES regard to animal growth hormones. Thus, gene was transmitted to subsequent genera- beginning in the early 1970s, extraction tions of progeny. techniques were developed to recover various Much effort in industrialized countries is growth hormones from their respective ani- being devoted to producing transgenic fish. mal pituitary glands, including several fish Fish are good candidates for study for several species. Since each of these procedures was reasons: the large, transparent and externally laborious and costly, very little of these fertilized eggs of fish species make them "natural" growth hormones were available to ideal subjects for genetic manipulation; the researchers. After techniques were developed embryonic development may be relatively to construct genetically engineered bacteria easily observed and studied in fish eggs; and that produce growth hormone, the Maryland fish that have been genetically altered may be Biotechnology Institute's Center of Marine more valuable commercially than their un- Biotechnology genetically engineered E. coli treated relatives (Liu and others 1990). to produce rainbow trout growth hormone There are three stages to developing a (tGH). In subsequent experiments, the geneti- transgenic fish. First, the foreign gene has to cally engineered tGH was injected into year- be integrated into the genome of the target ling trout. The growth rates and increase in organism. The present method of choice for weight and length of the treated trout was delivering the foreign gene is by micro- markedly higher than of the untreated fish injection (Chen and Powers 1990). In brief, (Agellon and others 1988). Other experiments the foreign gene that is to be inserted into the involving the injection of recombinant salmon new host is cloned to produce a large number GH into rainbow trout and recombinant tuna of that gene. Then, a gene construct is devel- GH into the Japanese snapper produced oped by attaching a promoter to the gene, similar results (Heyward and Hammond and hundreds of the gene construct are inject- 1990). Although the results from these exper- ed into the nuclei of the target organism's iments would seem to hold promise for com- embryo by glass micro-pipette. The treated mercial opportunities, technical problems embryo is re-implanted into the female fish. associated with injecting large numbers of After the embryo is full-grown, DNA is fish with GH are so severe as to negate this extracted from it and analyzed. Individual approach in aquaculture. Alternative methods transgenic fish are identified and propagated had to be developed to raise the GH level in for further study. According to a recent fish. review, nineteen experiments have so far One way was to insert the genes coding been carried out to develop transgenic fish for GH production directly into the genome (Chen and Powers 1990). of the targeted organism. When this was Second, the gene must express itself done, it led to a spectacular manifestation of through the production of GH. Proof of applied molecular genetics-the development expression comes from the analysis of the of transgenic animals. These are animals into GH found in the fish and by comparing the whose genome a gene or genes from another size and weight of transformed fish with organism has artificially been introduced. untreated control groups. Referring to the Animals that have been successfully trans- nineteen experiments mentioned above, ten formed by the time of this writing include the different fish (common carp, Chinese carp, fruit fly, sea urchin, fish, frog, mouse, pig catfish, goldfish, loach, medaka, salmon, and cow. In the animals where gene transfer Tilapia, rainbow trout and zebrafish) have techniques have been particularly successful, been successfully transformed (Chen and including several species of fish, the foreign Powers 1990). Marine biotechnology and Its sub-areas 27 Third, the new trait has to be transmitted pleuronectes yokohamae, which inhabits the to subsequent generations of progeny. The Yellow Sea. After having isolated and charac- trait in progeny is detected in the same man- terized the gene coding for this protein, these ner as in the foregoing step. Considering the scientists were able to synthesize this gene ten fish wherein expression was achieved, in and insert it in the bacteria E. coli, which only two cases was the trait successfully manufactures large quantities of the protein transmitted to progeny (Chen and Powers (Yaoqing and Xiongfeng 1990). 1990). In addition to the safety concerns that The progeny from one of the two success- surround the development of transgenic fish ful experiments are now being field tested. (and other transgenic organisms), difficult Researchers at Auburn University in Alabama scientific and technical problems will have to (USA) are growing transgenic carp to repli- be overcome before this sub-area of marine cate under farm conditions the 40 percent biotechnology will progress significantly increase in growth observed in the laborato- (Kapuscinski 1990). Specifically, techniques ry. (The details of this test and the safety of for gene transfer have to be improved; better field testing are discussed in Chapter 4.) promoters that mediate the foreign gene will Research on transferring genes other than have to be found and tested; and physical, those that code for GH is also under way. chemical and environmental factors that For example, certain fish that live in ex- maximize the effectiveness of the transgenic tremely cold waters (between 0° and -2° C) fish will have to be determined (Chen and have evolved so they produce antifreeze Powers 1990). proteins, which prevent their blood from freezing. However, most food-fish, such as GROWrH AND REGULATION OF BIVALVES, salmon, salmonids and trout, do not have this GASTROPODS AND CRUSTACEANS. The major protein. The susceptibility of salmon to death groups of bivalves are clams, oysters, mus- by freezing has, for instance, prevented sels and cockles; gastropods include abalone; salmon aquaculture in Atlantic Canada (Cut- while crustaceans include shrimp, crabs and ler, Saleem and Georges 1989). Inserting the lobsters. Oysters are considered a delicacy by gene coding for the anti-freeze protein could many, and much research is directed at im- thus help in extending the living range of proving yields from oyster farming to satisfy important food-fish into waters colder than a growing demand. Fortunately, the culture those they now inhabit. Scientists at the Johns process provides a good opportunity to ma- Hopkins University in Maryland and the nipulate the animal's genome for increased University of Illinois have identified and disease resistance, faster growth, or triploidy. cloned the genes that code for the anti-freeze The process of culturing oysters begins proteins in the winter flounder (Gourlie and with the induced spawning of sexually mature others 1984). Attempts have been made to females and males, whose egg and sperm transfer these genes to the Atlantic salmon. unite to form embryos. Embryos develop in Although the genes were expressed, the level a short time in larvae, which grow until they of proteins was too low to afford protection reach a stage of maturity when they metamor- against freezing (Chen and Powers 1990). phose into juvenile oysters (spat). Spat usual- Work to improve expression of the genes is ly set on cultch, where they harden until final proceeding rapidly, however. Antifreeze maturation, which is called "grow-out." Har- proteins are also being studied in China, dening and grow-out can be induced to take where researchers at the Institute of Genetics, place on the seabed or in floating rafts. Spats Beijing, are investigating the fish Pseudo- grow to market size in two to three years. 28 MARINE IOTECHNOLOGYAND DEVELOPNG COUNTRIES There are several biotechnology tech- oysters that do not attach themselves to a niques that can be applied to improve the solid surface). The cultch-less oyster has two aquaculture of oysters. The application of one advantages over its wild relative. First, be- set of techniques, triploidy, deserves descrip- cause it grows much faster, it can be brought tion because it has had a remarkable econom- to the market faster, and the disease problem ic impact. is nearly eliminated since the oyster is har- Triploid oysters, which contain three sets vested before the full effects of disease be- of chromosomes instead of the normal two, come manifest. Second, the culinary quality were first developed by researchers at the of the cuitch-less oyster is high, making it a University of Maine in 1979. This accom- favorite for oyster lovers. In view of the plishment, which was based on earlier work cultch-less oyster species' qualities, an indus- done by scientists in Norway on fish, resulted trial firm specializing in mariculture has from treating oyster eggs with the chemical entered into a contract with the university to cytochalasin B, which inhibits normal cell market the oysters. division, creating two sets of chromosomes in Bivalves are usually not cultch-less, but the egg. Triploidy results from the union of instead their larvae settle on hard surfaces this egg with a normal sperm containing one where they attain their adult form and remain chromosome set (Allen 1988). the rest of their lives. The settling phenomena After oysters was deemed safe for human has been intensively studied by several consumption by the U.S. Food and Drug groups, but most scientists refer to the pio- Administration (FDA), the technique was neering research done in this subject by Dr. quickly applied by aquaculturists in the state Daniel Morse's group at the University of of Washington. The reason for their positive California at Santa Barbara (Morse and reception was that the farmed oyster in the Morse 1988). In a series of elegant experi- U.S. northwest, Crassostrea gigas, spawns ments involving abalone, this group showed during the summer and becomes less flavorful that spawning (the release of fertilized eggs because reproductive tissue forms throughout into the seawater) in this animal was induced the body. Triploid oysters, being sterile, do and regulated by prostaglandins (a group of not undergo seasonal change and thus main- hormones that regulate reproduction in ani- tain constant texture and flavor throughout mals). The synthesis of abalone prostaglan- the year. In addition, triploids are valuable to dins is controlled by the rather common aquafarmers because they grow faster and to inorganic chemical hydrogen peroxide. The a larger size than do diploids. Triploids have spawning of abalone is thus easily triggered become a marketing success story; they by adding small amounts of hydrogen perox- represented about 50 percent of the U.S. ide to the seawater surrounding the site where northwest's total production in 1988 (Allen researchers wish the event to take place. 1988). After spawning, the development, settling Understandably, the loss of oysters to and metamorphosis of the larvae depend on disease is a serious problem to aquaculturists. the recognition by the larvae of specific Oyster production in the Chesapeake Bay, for molecular signals. Unless this signal is re- example, has declined precipitously during ceived, the larvae will remain free swim- the last few years, due primarily to a combi- ming, to die within a month. However, if nation of pollution and disease. To meet this larvae swim in the proximity of certain red threat, an R&D project was begun in October algae that encrust rocks, they will settle on 1987 at the University of Maryland to devel- the algae or nearby surface. The chemical op and propagate cultch-less oysters (that is, that induces settling was found to be a pep- Marine blotechnology and Its sub-areas 29 tide (a short chain of amino acids), similar to Biotechnology-related research on crusta- a neurotransmitter chemical found in animals cean species lags behind that being done on called GABA (gamma-aminobutyric acid). finfish and bivalves. The most immediate Addition of the inexpensive GABA to sea- applications of marine biotechnology to water will induce larvae to settle on specified crustacean aquaculture relates to enhancing surfaces. animal health. Morse's work has led to other investiga- tors researching similar phenomena among GROWITH AND REGULATION OF ALGAE. giant clams, mussels, oysters, scallops and Algae are nonvascular, photosynthetic plants other bivalves. The aggregate of research has that contain chlorophyll (Robinson 1985). led to an explosion in the knowledge about They may range in size and complexity from the molecular mechanisms of sensory recep- the microscopic, single-cell to the 70-meter tors, gene expression and cellular response to giant kelp Macrocystis. It is important to note chemical stimuli, and signal transmission in that marine plants, like their terrestrial coun- neural networks (Morse and Morse 1988). terparts, are primary products of Findings are also being applied by industry. nature-consuming CO2 absorbed in seawa- For example, abalone has been in great ter, using sunlight as their sole source of demand in California as a culinary delicacy, energy, and requiring little additional input in driving up prices and stimulating overfishing terms of trace nutrients. Thus the production of the gastropod. At the same time, the of plant biomass or natural substances by protected marine otters have grown in num- plants require lower levels of support energy ber from a few dozen to over 5000. Unfortu- than do similar production by bacteria, yeast nately, otters' preferred food is abalone, so or animals, which are secondary products of where otters live and propagate, abalone soon nature, feeding on plants, animals or fossil disappear; and California abalone has become fuels. a rare treat purchased at a very high price. Algae are generally classified according to To meet the market demand, three companies two groups; macro- and micro-algae. There have in the past five years or so begun to are over 21,000 macroalgal species in the culture abalones, using hydrogen peroxide to world, but the most numerous are Rhodo- bring on spawning and GABA to induce phyceae, or red algae (accounting for more settling (Morse 1991). Other bivalve pro- than 60 percent of these species); Phaeophy- grams utilizing similar techniques have been ceae, or brown algae (25 percent); and Ctlo- established in California; abalone and bivalve rophyceae, or green algae (about 15 percent). aquaculture capitalizing on this research is Red and brown algae are commercially im- also well under way in China and Taiwan. portant to the colloid-using industry (Renn Also of commercial importance is research 1986b); red and green algae are important being undertaken at the University of Mary- food in Asia. Algae may be collected from land to define a "gene bank" for the commer- wild stocks, primarily giant kelp, or grown in cially important oyster Crassostrea virginica aquaculture. The total world production of (Colwell 1986). This accomplishment sets the macroalgae per year is approximately 4 stage for the manipulation of this species' million tons worth about $1 billion. The genome. Oysters are fine experimental sub- largest producers are China, Japan and the jects since they produce a very large number Republic of Korea (Ruying and Qinguin of larvae and intermediate stages, allowing 1992), but significant quantities of wild scientists to easily and quickly observe the seaweed are harvested in California, the effects of their trials. eastern United States and Canada, Chile, 30 MARINE IOTECHNOLOGY AND DEVELOPING COUNTRIES France, the United Kingdom, Indonesia and fuel digesters, which produce hydrogen for the Philippines. local use as energy (Gold and Shultz 1986). Of more pertinence to this report, algal Green and red algae are used for food in aquaculture has been practiced in Asia for a Asia; for example, algae of the genus Por- long time and has much importance for phyra is used to produce the food nori. Nori producing food, fertilizer and industrial is an important food staple; in 1986 9 x 109 chemicals. Two algal aquaculture techniques sheets worth $450 million were produced for predominate. The first is outplanting, which the Japanese market alone (Japan 1991). is the culturing of algae in the ocean waters However, the major industrial importance of of the coastal zone. One outplanting tech- algae is that they contain a group of chemi- nique consists of suspending ropes impregnat- cals called hydrocolloids, which includes ed with spores from rafts. The spores develop agars, algins and carrageenans. In 1989 the into plants, which are allowed to develop worldwide demand for alginates (from mainly naturally. Ropes may be raised or lowered to brown algae) was satisfied with the produc- expose algae to optimum light, accelerating tion of 35,000 tons; most of it was used by normal growth by several months. In another the textile industry (50 percent), food indus- technique, cuttings of the alga are tied to a try (30 percent), paper industry (6 percent) monoline stretched along the seabed. Harvest- and pharmaceutical industry (5 percent). The ing takes place after two or four months, production of carrageenans (from red algae) after the plant has attained a weight of more was 17,000 tons, while the world's demand than I kilogram (Llana 1991). Outplanting is for the substance stood at 20,000 tons (Rich- practiced extensively in China, Japan and the ards-Rajadura 1990). Carrageenans are used Philippines to cultivate various types of red by the food industry (78 percent) and cosmet- alga. ic industry (22 percent). The production of The second aquaculture technique is grow- agars (from red algae) was 6,700 tons; this ing algae in a closed system. In this method, quantity is insufficient to meet world demand, large ponds are dug into muddy ground and which grows 25 percent each year (Polysac- the ponds formed are subdivided into com- charides 1991). Agars are used by food partments of about 0.2 hectares. Each com- industry (58 percent), scientific laboratories partment has an entrance and exit gate to (28 percent) and pharmaceutical industry (14 facilitate flooding. Seawater is let in and percent) (Mabeau, Valiat and Brault 1990). cover the ponds to a depth of about .8 me- Agars are particularly important to the con- ters. Algal cuttings are stuck in the mud and duct of biotechnology research and to bio- allowed to grow for about two months or industry processes. until the bottom is uniformly covered (Llana The processing of algae by industry to 1991). This method demands a constant extract and purify hydrocolloids consists of supply of fresh, unpolluted seawater and washing the algae; dissolving the raw hydro- requires the addition of nitrogenous fertilizer. colloids with alkali; clarifying and precipitat- The technique allows the farmer to control ing the hydrocolloids; removing unwanted that part of the growth cycle when algae are color; purifying the compound by ion ex- growing the fastest and it eliminates losses change; drying and milling the hydrocolloid from inclement weather. Closed systems are (Mabeau, Valiat and Brault 1990). Algae used by Europeans, Filipinos and Taiwanese processing is a low to medium technology to cultivate species of red algae. endeavor to which biotechnology would seem Algae have several uses. Bulk macroalgae to have little application. However, biotech- is used by Asian farmers as fertilizer and to nology techniques may be applied to develop Marin blotechnology and Its sub-areas 31 algal strains that possess more favorable since low-cost protein from terrestrial crops characteristics than do wild strains. For is abundantly available. To illustrate, the cost example, researchers at Northeastern Univer- of efficiently producing one kilogram of sity in Boston (USA), are developing hybrid Chlorella is about $10, while Spirulina costs strains of agar-producing macroalgae that can $2-$10 per kilogram. In comparison, the be cultivated more efficiently than the wild high protein-containing soybean costs only strains and that produce more agar per unit about $0.2 per kilogram (Chapman and weight. The widespread introduction of Gellenbeck 1989). Reflecting its high price, genetically improved strains may lead to a in the food industry microalgae has a limited decrease of the cost of agar, which now is market as health foods, but little else. Even if $24-$200 per kilogram, and agarose, which the production costs of microalgae is dramati- costs $250-$40,000 per kilogram (Renn cally lowered, regulators will have to deter- 1991). Other strains could be developed that mine whether dried microalgae is safe when contain increased amounts of specialty chemi- consumed in large quantities. Appropriate cals and pharmaceuticals. It is interesting to testing will have to be done to settle this note that while 80 percent of the macroalgae question. destined for commerce originate in the devel- Beyond food, microalgae can be used to oping countries of the Asia-Pacific region, 90 manufacture substances of interest and value percent of the colloid industry is located in to industry. For example, food industry and West Europe, Japan and Korea (Ruying and mariculture needs large quantities of pig- Qinguin 1992). ments, such as beta carotene, phycoerythrin An illustrative example of what is being and zeaxanthin. One successful production done in algal biotechnology is the work on system has been developed by Microbio Re- macroscopic alga at the University of North sources, San Francisco (USA), consisting of Carolina Center for Marine Science Re- large open outdoor tanks or ponds populated search. Researchers are using cell culture by Dunaliella, which synthesize large quanti- techniques to enhance the growth characteris- ties of the Vitamin A precursor beta carotene. tics or red alga for the purpose of increasing Similarly, the Israeli firm Koor Foods is this species' ability to produce agar. An developing Dunaliella bardawilli, a species of interesting aside is that parallel work, using microalgae found in the Sinai peninsula, for the same techniques, is proceeding on angio- glycerol and beta carotene production. Anoth- sperm (a plant, such as eelgrass, that grows er Israeli group at the Ben Gurion University well in shallow waters). The objective of this is investigating the possibility of using Por- work is to enhance the plant's ability to grow phyridium to produce on a large-scale a close to shore so it will be able to hold sand carrageenan-like polysaccharide in large out- dunes in place and to stabilize mud flats door salt water ponds (Weiner 1985). (CMSR 1990). Although the classical techniques of muta- Microalgal R&D has been focussed mostly tion, selection and breeding are most com- on four species; CZlorella, Dunaliella, Scene- mon to microalgae R&D, advanced biotech- desmus and Spirulina. The approach used is nology techniques are being used to develop similar to that in industrial microbiology, strains that will grow faster, be more resistant namely mutagenesis and selection of mutants to disease, and better adapted for mass propa- with desired traits. The aim of Chlorella and gation (Tucker 1985). Similar to the ap- Spirulina R&D has been to develop these proaches researchers use when working on organisms for low-cost, high-protein food terrestrial bacteria, fungi and higher plants, production, but this goal has remained elusive certain problems have to be overcome in 32 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES microalgae biotechnology. First, the foreign processing gene has to be successfully introduced into * produce new or unique products the microalgae. A major problem here is to * are more disease resistant than present find ways to penetrate the tough microalgal species cell wall. Second, the entry of the gene into * will grow in colder, warmer, shallower or the new host has to be monitored as does its deeper water than is now possible ability to express the desired product. Third, * possess better nutritional characteristics the stability of the introduced gene has to be than present species ascertained; that is, the introduced material * produce more biomass per cubic unit must be incorporated in the host's genome (Renn 1986a). where it must continue to function as expect- ed. In addition, when the microalgal cell Marine animal health divides, the gene must be replicated and passed on in subsequent generations of proge- Bacterial, fungal, protozoan and viral ny. Although encouraging progress is being infectious diseases are widespread among reported, scientists have not solved these natural fish populations. But animals raised in three problems (Brown, Dunahay and Jarvis intensive aquaculture are especially vulnera- 1989). ble to damage by disease. The most common Nevertheless, respectable progress is being bacterial disease agents found in fresh water achieved in microalgae biotechnology. For aquaculture are Aeromonas and Pseudomo- example, a team at the Solar Energy Re- nas; while in mariculture the major pathogen search Institute in Colorado (USA) is apply- is Vibrio. Polluted waters favor the develop- ing rDNA to isolate genes that regulate lipid ment of fungal diseases; the most common biosynthesis and to introduce these genes into agent is Saprolegnia (Shariff and Subasinghe microalgae that have potential for outdoor 1990). Two types of viral diseases are partic- mass culture (Brown, Dunahay and Jarvis ularly deadly to aquacultured salmon-infec- 1989), while the Australian company Wes- tious pancreatic necrosis and infectious hema- farmers Algal Biotechnology's success in topoietic necrosis (IHN). Similar diseases developing microalgae for the production of afflict catfish, flounder, menhaden and striped beta carotene and animal feed. Other micro- bass (Klausner 1985). Protozoans, mainly algae production systems are being developed ciliates and flagellates, damage fish by feed- to produce amino acids, animal feeds, fatty ing on and within their skin and gills (Noga acids, feed pigments, hydrocarbon fuels, 1987). The Epizootic Ulcerative Syndrome is pharmaceuticals and polysaccharides (Bene- a widespread disease afflicting fresh and mann 1989). brackish water fish; the causative organism is To sum up, whether macroalgae or micro- unknown. Bacterial, fungal and viral diseases algae, the advanced techniques of biotechnol- also afflict bivalves and crustaceans. ogy could be employed to improve the algae In general, waterborne bacterial and viral itself, making it more useful to industry. pathogens are attenuated in well-managed Theoretically, these improvements may devel- aquaculture ponds and basin, probably due to op varieties that: the pH of the water and predation by proto- zoans and zooplankton (Edwards 1991). * grow faster However, when fish do become afflicted by * yield higher concentrations of desired bacterial diseases, aquaculturists will as a products matter of course disperse antibiotics, anti- * contain fewer impurities that complicate bacterials and disinfectants in the waters of Marine blotechnology and Its sub-wreas 33 ponds and tanks holding the diseased fish. aquaculturists. Only two efficacious vaccines The quantities of antibiotics used in aquacul- are, however, available for fish and one for ture is large, for example, in Norway, which lobster; none is available to prevent diseases is the world's largest producer of aqua- afflicting shrimp and shellfish. cultured salmon, aquaculturists used 17,000 Whether a scientist is attempting to devel- kilograms of antibiotics in 1985, but in- op a vaccine for use in terrestrial animals or creased this amount to 48,000 kilograms in marine animals, R&D methods will be simi- 1987 (Technical Q&A 1991a). Antibiotics lar. In general, there are three types of vac- commonly used in aquaculture include amino- cines; attenuated, killed and biosynthetic glycosides, beta lactams, tetracyclines, vaccines. Attenuated vaccines consist of macrolides and chloramphenicol. Most of pathogenic organisms that have been treated them are also used to treat infectious diseases so they are no longer infective, but are still of humans. The employment of antibiotics in able to elicit an antibody reaction in the aquaculture poses potentially serious public vaccinated host. The attenuated vaccine is health and environmental problems since somewhat more risky than the others because genes coding for antibiotic resistance may the attenuated organism may under certain disperse via conjugation from wild bacteria circumstances revert back to the infectious affected by aquaculture operations to human form capable of causing disease. The second pathogens, such as the bacterial species type of vaccine consists of the killed patho- causing typhoid fever, cholera and dysentery. genic organism. Although safe, this vaccine Further, if improperly used, residues of type may elicit a weaker antibody response in antibiotics may taint the flesh of harvested the host than do the others. The third vaccine fish. Proper management practices can lessen type is one developed via genetic engineer- risks associated with antibiotic usage in ing. Briefly, a protein constituting the patho- aquaculture, but cannot eliminate it altogether genic bacteria's cell wall or the virus' surface (Technical Q&A 1991a). that is capable of eliciting an antibody re- T'here are no approved drugs for treating sponse in the host is identified. The gene fish viral diseases (although aquaculturists coding for that protein is constructed, then have over time empirically developed a inserted in an industrial bacterium, which variety of methods for containing diseases produces large quantities of the protein. The afflicting their stocks). Since fish that survive protein then becomes the basis of an effec- a viral disease outbreak may become carriers tive, safe vaccine. of the causative virus, once certain diseases Those who seek to develop fish vaccines infect a pond, the only recourse may be to are faced with two problems that their coun- destroy the animals it contains and to empty terparts working on terrestrial animal vac- and decontaminate the pond itself. These cines do not have to contend with. First, the drastic steps obviously are costly and may causative organisms of most of the diseases bankrupt the aquaculturist. To illustrate, affecting marine animals are unknown, there- shrimp production in Taiwan dropped from fore much basic research is required to identi- 114,000 metric tons in 1987 to about 50,000 fy and characterize infectious agents afflicting metric tons in 1988 and 30,000 tons in 1991 marine animals and to clarify the intricate (Record year 1992), largely due to the ravag- relationships between hosts and parasites. es of a disease that decimated black tiger Second, fish being what they are, are difficult shrimp. The ability to prevent by vaccination to vaccinate. There are two methods now in diseases that afflict animals being cultured use. First, fish may be immersed in waters would undoubtedly be of immense value to containing a high concentration of the vac- 34 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES cine. Second, fish can be vaccinated by efficacy, safety and price. This is being injection. Each method presents difficulties, developed by scientists at the Oregon State principally with the dilution of the vaccine University, who have identified, characterized past usefulness and the stability of the prepa- and cloned several genes coding for proteins ration under harsh physical conditions. The that elicit antibody formation in fish. High immersion method is cost-effective, but some levels of the protein they code for have been vaccines will not work when administered expressed (Engleking and Leong 1991). The this way. Vaccinating fish individually is a next step is to scale up the manufacture of manpower-intensive exercise; its costs cannot candidate vaccine and to gain approval from be justified unless the fish to be vaccinated the USDA to test it in the field. are relatively valuable (for example, food-fish Substances other than vaccines may have that are near to market size and fish valued protective functions. For example, an extract by collectors). It bears mentioning that a new from the shellfish Ecteinascidia turbinada method for administering vaccines (and protects eel from Aeromonas infection and in therapeutics) to fish is being tested. It utilizes general enhances the immunological defenses a combination of immersion and ultrasound. of blue crab, crayfish and prawn (Colwell Fish immersed in tanks that are exposed to 1986). The Phillips Petroleum Company ultrasound for 10-15 minutes record a 10-20 Norway sells a glucan, produced by yeast, fold increase over controls in internal levels with the trade name Macrogard, which it of the administered drug (Gain 1991). claims improves the efficiency of vaccines Of the ongoing work to develop fish and helps farmed fish resist disease (Hoffman vaccines, that being done on a vaccine against 1990). Undoubtedly a host of as yet undis- IHN may be furthest along. The importance covered biological substances from marine of this viral disease was recognized in 1953 organisms have antibiotic, protective and when it caused a massive die-off of salmon in curative powers that will benefit aquaculture. the state of Washington. The disease has Some R&D is directed at detecting fish since that time spread as far as Japan, causing diseases. Diagnostic methods based on mono- damaging epidemics in many salmon and clonal antibody technology (see below) are trout hatcheries. Wild fish are not spared; an being developed at the University of Maine estimated 20 percent of salmon fry in British for infectious pancreatic necrosis and IHN Columbia die from this disease (Powers (Klausner 1985). Diagnosing these diseases at 1990). In view of the damage this disease an early stage may help in limiting loss of causes, it is understandable that much effort fish stock, as carriers of the viruses causing is being put into investigating the IHN virus the diseases can be identified and eliminated, and developing a vaccine against it. In fact, limiting the spread of diseases. In addition, prototypes of the three types of vaccines have better diagnostics will be useful in vaccine been developed and tested in the laboratory R&D. (Powers 1990). All protected fish against IHN virus when injected. However, the first Marine natural products (a conventional type killed vaccine) proved not useful when administered in water. The Natural resource chemists have for over second, the attenuated type, was effective via 100 years been screening the world's organ- water borne inoculation, but questions regard- isms for useful chemical substances. The ing its safety have not been resolved. The results of this effort is impressive; about third type, which is a recombinant killed type 20,000 chemicals from natural products have vaccine, shows most promise in terms of been characterized and the annual sales of Marlne blotechnology and its sub-areas 35 pharmaceuticals derived from plants reaches cetes, are especially prolific producers of $10 billion per year in the United States antibiotics. In fact, 74 percent of the antibiot- alone. Yet little or nothing is known about ics produced by the pharmaceutical industry the chemical composition of most existing come from actinomycetes. However, numer- terrestrial plants and microorganisms; those ous other bacteria and fungi produce thou- inhabiting the marine environment are even sands of different antibiotics. By far most more alien. It is no surprise, therefore, that antibiotics, however, have limited or no large numbers of new natural substances are utility to humans because they are too toxic, discovered every year. unstable, or possess other undesirable charac- By and large, chemicals constituting teristics. organisms fall within one of two groups. A relatively large number of marine First, there are primary and intermediate organisms are known to produce secondary metabolites and cofactors, which are essential metabolites that possess antibiotic properties, for the growth of the organism and its repro- including blue-green, green, brown and red ductive metabolism. Second, there are sec- algae, dinoflagellates, sponges, jellyfish, sea ondary metabolites-substances that have no anemones and others (Baslow 1977). So far essential function and whose reason for little research has been done on groups of existence are not so clear. They are thought organisms and the specific antibiotics they to have arisen in the course of evolution in produce. One exception to this rule came order to confer on the organism a particular about as a result of an investigation of the advantage, which is now unclear (Vining predator-prey interactions between a nudi- 1991). Secondary metabolites have diverse branch and sponge. The nudibranch, which is chemical structures, but smaller groups of soft-shelled and slow moving, appears vulner- organisms, such as genus, species, or even able to predators. Yet, it is but rarely at- strain, often produce substances distinct to tacked. Upon investigation it was found that that group. Since secondary metabolites are the animal secretes an antibiotic substance, by far the most important group of useful which scientists eventually isolated and identi- chemicals produced by marine organisms, fied, then named mimosamycin (Scheuer and since their evaluation is in fact the objec- 1990). Mimosamycin belongs to a chemical tive of marine natural product research, group named isoquinolinequinones, which discussion here is limited to these chemicals. had first been found a terrestrial Streptomyces Secondary metabolites, whether of marine species. Further research is required in order or terrestrial origin, do not have a single to clarify if mimosamycin is in fact secreted function but instead exhibit a wide range of by the nudibranch itself or by a commensal activities. Five types of activities are impor- Streptomyces species colonizing the nudi- tant for our consideration: antibiotic; anti- branch. inflammatory, anti-tumor and anti-viral; marine toxins; enzymes; and secondary ANTI-INFLAMMATORY, ANTI-TUMOR AND metabolites as insecticides and herbicides. ANI-viRAL. As mentioned, most antibiotics cannot be used in animals, including humans, ANTIBIOTIC. Microorganisms, whether in because they are too toxic. The toxicity of marine or terrestrial environs, face intense these agents stem from their actions being competition from other organisms. Some are nonselective in that they interfere with uni- able to gain an advantage by producing anti- versal metabolic reactions in the recipient biotics that kill or inactivate competitors. (Vining 1991). Some of these toxic agents Certain soil bacteria, particularly actinomy- affect primarily rapidly dividing and growing 36 MARINE BIOTECHNOLOGYAND DEVELOPING COUNTRIES cells. By adjusting dosage and directing the whether the sponge itself is the source of action of these agents, they can be useful in these compounds or different microorganisms treating tumors. Two such agents are now in that live in a commensal or symbiotic rela- clinical trials at the NCI. The first is Bryosta- tionship with the sponge. Studies of sponges tin, which was isolated from the bryozoan reveal that an extremely large number of Bugula neritina, which lives in a symbiotic anaerobic bacteria, cyanobacteria, dinoflagel- relationship with a yellow sponge found only lates, heterotrophic bacteria, and microalgae in the Gulf of California, Mexico. The sec- inhabit them. When tested, some of these ond is Didemnin B, isolated from the Carib- microorganisms proved to possess bioactive bean tunicate Tridedemnum solidum. Didem- properties. These findings clearly show that nin is active against leukemia and melanoma; much research has to be done to clarify the in addition, it shows strong anti-viral and complex interrelationships between macro- immunosuppressive activity. Didemnin is in organisms, such as the sponge, and the mi- clinical testing (Klausner 1986). A n o t h e r croorganisms that colonize them. secondary metabolite having promising prop- Another problem with trying to determine erties is manoalide, which is named after the the origin of a bioactive compound is that Manoa Valley in Oahu, Hawaii. This com- some marine organisms, which lack physical pound was found by a University of Califor- protection such as a shell, use chemicals to nia at Santa Barbara researcher who had been deter attackers. However, the defensive investigating marine sponges for more than chemicals are not always produced by the thirteen years. Manoalide was isolated from organism wielding them but instead are the South Pacific sponge Luffariella varia- derived from exogenous sources. For exam- bilis, and belongs to a group of chemicals ple, nudibranchs commonly feed on inverte- called terpenes, although its structure is brates. Some nudibranchs appear to sequester highly unusual (Klausner 1986). Detailed from their prey substances toxic to fish that pharmacological studies of manoalide has are thereupon stored in its tissues (Klausner shown it to have powerful anti-inflammatory 1986). This protection may even be passed and analgesic properties, as well as evidenc- on. The nudibranch "Spanish dancer' depos- ing some anti-leukemic and anti-fungal prop- its its eggs in the open, on rocks and corals. erties (Austin 1989). In view of these find- Because they contain the bioactive substances ings, the pharmaceutical company Allergan called ulapualides, the eggs are free from Corporation entered into a joint venture with predation. Incidentally, ulapualides exhibit manolide's discoverer; the substance is cur- anti-leukemic and anti-fungal properties. rently in clinical trials for use against skin disorders, includingpsoriasis. Further investi- MARINE TOXINS. Some secondary metabo- gation of the same sponge has led to the lites do not display general cytotoxicity but discovery of two other compounds, which are have specific pharmacological activity that named Luffariellin A and B. Initial testing make them extremely toxic to animals. For indicates that these compounds also possess example, saxitoxins derived from dinoflagel- anti-inflammatory properties. lates are fifty times as potent as curare In general, sponges have proven to be a (Rodrigue and others 1990). Since food good source of secondary metabolites. The chains in the marine environment often begin hundreds of compounds have been isolated with dinoflagellates, it is no surprise that from sponges represent many classes of much human suffering has been caused by chemicals, including alkaloids, sterols, terpe- marine toxins. In 1987, for instance, 26 noids and others; some possess unique struc- people died and another 161 were affected in tures (Crews 1991). What is not so clear is Champerico, Guatemala after they ate clams Marlne biotechnology and fts sub-areas 37 that had assimilated saxitoxins from dino- kill humans. The toxins act on calcium chan- flagellates (Rodrigue and others 1990). Other nels and sodium channels, as well as being epidemics of paralytic shell fish poisoning targeted to neuromuscular and vasopressin have occurred recently in Borneo, Indonesia, receptors (Olivera and others 1990). Japan, New Guinea, Palau and the Philip- Much remains to be found out about pines. marine toxins before use can be made of Perhaps even more frequent than shell fish them in medicine or industry. One can imag- poisoning is ciguatera fish poisoning. The ine, for example, that palytoxin, which is a sequence of events that take place before man nonprotein polypeptide having approximately is poisoned by this toxin is unmatched in the same degree of toxicity as ricin (a toxin nature. The ciguatoxin is produced and re- found in the castor bean), may find similar tained by dinoflagellates, which eventually uses as this toxin. Ricin, which is exceeding- settle on a macroalgae. Herbivorous fish feed ly cytotoxic, has been coupled with mono- on the macroalgae, incidentally ingesting the clonal antibodies programmed to attach them- settled dinoflagellates. Predator fish eat the selves to the cells of certain cancer tumors. herbivorous fish, and then in turn are caught Once so attached, the ricin selectively kills and eaten by people. The toxin thus passes that cell without harming normal tissue. unchanged through four different hosts in Palytoxin, and other marine toxins, may order to harm the fifth (Scheuer 1990). work better than ricin on certain cancers or When appropriately dispensed many toxins may have other advantages that cannot now are valuable medicines as analgesics and be determined. As one scientist has noted, muscle relaxants; others have anti-tumor "Peptides [toxins] are primary translation activity. Thus the marine sponge Haliclona products of genes, with potent biological produces halitoxin, which inhibits the growth activity, and these peptides can be manipulat- of certain types of tumors. Toxins that cannot ed by the techniques of modern molecular by used as medicines because of side-effects genetics. This dual quality confers on the or other problems nevertheless have impor- [toxins] an important role in the expanding tant uses as models for the design and synthe- bridge between chemistry and modern molec- sis of other drugs (Colwell 1986). Toxins are ular genetics."(Olivera and others 1990) also exceedingly valuable to medical Marine toxins may pose difficult challeng- researchers who study nerves and nerve es to science. For instance, natives of the impulse transmission, the central nervous Hawaiian island of Maui when fending off system, and smooth muscle action. For exam- invaders used a deadly toxin to tip their ple, tetrodotoxin, which is produced in spe- spears. The source of the toxin was, and cialized glands of the puffer fish and by remains, tidal pools inhabited by marine certain marine bacterial species, acts to para- animals called Palythoa, which includes lyze the peripheral nerves. It is used in re- corals, anemones and jellyfish. However, search to elucidate the nerve excitation mech- these organisms do not make the toxin; it is anism. Lophotoxin, which comes from the manufactured by a marine bacteria that lives gorgonia Lophogorgia, inhibits nerve-stimu- in symbiosis with Palythoa. Intensive investi- lated contraction of muscle (Colwell 1986). A gation is proceeding to clarify how these bac- bacterium living commensally in the digestive teria make the toxin, the toxin's chemical gland of the shellfish Babylonia japonica structure, and its deadly mode of action (Fox produces neosurugatoxin and prosurugatoxin, 1982). which are powerful ganglion-blocking agents (Vining 1991). The toxins found in the preda- ENZYMES. Enzymes are used as pharma- tor cone snail of the Conus genus can easily ceuticals, food additives and fine chemicals. 38 MARINE BIOTECHNOLOGYAND DEVELOPING COUNTRIES But in nature, enzymes are chemicals vital to of high heat, tremendous pressure, no light life, catalyzing metabolic reactions, breaking and high salinity. A profusion of such organ- down waste products, and making possible isms are found inhabiting the abyssal depths the transmission of neural signals. Marine of the ocean proximate to thermal vents. For organisms are richly endowed with many example, marine bacteria of the Archae- enzymes; some are unique. An example of a bacteria genus, giant tube worms, and others unique enzyme having enormous industrial live and propagate at a depth of 2,000 meters potential is one being developed by a and more, and where the temperature is 950 researcher at the University of California at C and higher. Much research is being per- Santa Barbara. Dr. M. Polne-Fuller was formed to clarify how proteins, nucleic acids performing basic research, investigating the and enzymes are able to function at these relationship between brown alga and an temperatures, which kill most organisms. amoeba of the Trichospaheriwn genus. In the Practical applications from this research are course of this work, she noted that the amoe- already being realized. For example, CP ba had the ability to dissolve the alga. Since Laboratories in England is marketing an the alga contains many chlorinated and bro- enzyme called Vent DNA polymerase, which minated compounds, she reasoned that has been purified from Thermococcus litora- through an evolutionary process, certain lis, a type of archaebacterium. This enzyme amoeba had gained the ability to digest these is useful in certain laboratory reactions be- compounds. Plastics, which are high molecu- cause it remains active for over two hours at lar weight polyethylenes and polyvinyls, are 1000 C (hermostable 1990). Soon it should typically chlorinated and brominated. Dr. be possible to utilize these findings in manu- Polne-Fuller wondered whether the amoeba facturing processes catalyzed by enzymes. would perchance attack plastics. When tests These processes, when utilizing conventional indicated that the amoeba would indeed enzymes, tend to be inactivated by heat degrade plastic, she initiated research to higher than about 40° C. Since enzymes from improve the amoeba's degrading ability. extremophiles will have greater stability and Using mutation by ultraviolet light and selec- last longer at high temperatures than those tion, Polne-Fuller eventually isolated a mu- now used by industry, their availability would tant amoeba strain that has a powerful plastic open up new possibilities for efficient manu- degrading ability. More testing indicated that facturing (Tucker 1985). the mutant amoeba would destroy plastic in the field as well as in the laboratory. At this SECONDARY METABOLITES AS INSECTI- point the university applied to patent the CIDES AND HERBICIDES. Screening of terres- mutant amoeba and contacted industry about trial microorganisms and plants has led to the joint-venturing. Occidental Chemical Compa- discovery of many secondary metabolites ny has entered into such an agreement and is having insecticidal activity. Their actions now funding research to clarify the metabolic vary widely; they may block cellular respira- pathways of the amoeba by using C,-labelled tion, inhibit protein synthesis, interfere with polymers; these polymers are not available so the synthesis of chitin (which is a major they have to be developed before research constituent of insects' outer shells), or ob- proceeds to the next stage (Polne-Fuller, struct digestion. Undoubtedly a large variety Rogerson and Gibor 1991). of marine organisms produce secondary meta- An exceedingly promising area of marine bolites that have similar properties, but so far biotechnology is the study of extremophiles- little has been done in this area. For example, organisms that live under extreme conditions fishermen have known for a long time that Marine biotechnology and its sub-areas 39 flies settling on the annelid Lwnbrineris produce more chitosan and become dormant. brevicirra used as bait would die. Acting on So, perhaps, chitosan applied to plants may that observation, a crude compound contain- trigger this dormant condition. Second, chito- ing the active ingredient was prepared from san may activate disease resistance in plants the annelid as early as 1934. Eventually by stimulating genes' coding for enzymes that chemists elucidated the toxin's structure and digest fungal cell walls. named it nereistoxin. A synthesized derivative Chitin, which is the major component of called cartap hydrochloride is now marketed crustacean and insect shells, is the second as an insecticide. It is active against several most plentiful organic compound on Earth insect pests, including the rice stem borer. (after cellulose). It is usually considered a Unlike most chemical insecticides it is not waste-product by industries that process toxic to warm-blooded animals and insects do crabs, shrimp and other crustaceans. An not readily develop resistance against it estimated 150,000 tons of chitin is generated (Colwell 1986). per year as waste. Japan is the major proces- Screening of terrestrial organisms has also sor of chitin and has the largest supply of the uncovered several that produce secondary substance. However, there is an overabun- metabolites affecting plants, including plant dance of chitin in the world because so far hormones, wilting agents, growth regulators few applications for it have been developed. and phytotoxins. Some secondary metabo- lites, such as homoalanosine produced by Both research institutes and private com- Streptomyces galilaeus, have both insecticidal panies undertake or sponsor wide-ranging and herbicidal properties (Vining 1991). As programs for screening marine organisms for with the case of insecticides, we can also bioactive secondary metabolites. Screening expect to find marine organisms that produce assays may utilize tumor cell lines, DNA and metabolites which act on plants in differing RNA viruses, pathogenic bacteria, or a com- ways, including herbicidal effects. But so far bination of the three. As of this writing, more this area is mostly unknown. than 2000 secondary metabolites showing As an aside, certain properties of chitin bioactivity have been isolated from marine and chitosan have pertinence to this section. organisms. An illustrative example of what is Both substances are biodegradable and are involved in screening is a project funded by compatible with human body tissue. Chito- the U.S. National Cancer Institute (NCI) but san, a derivative of chitin, is carbohydrate being carried out by the Australian Institute with a simple chemical structure. It is poten- of Marine Sciences during 1989-91. The cost tially useful as fibre and film. More to the of the project is $1.05 million per year. The point, it has fungicidal and pesticidal proper- project's objective is for the contractor to ties useful to terrestrial agriculture. In experi- collect 1000 species of invertebrate organisms ments, chitosan stopped the growth of fungi and mollusks per year and submit the speci- pathogenic to plants, made pea tissue resistant mens to the NCI. A sufficient quantity of to invasion by fungi, and helped wheat culti- each organism has to be collected so 1 kilo- vars resist the effects of stem-rotting disease gram remains after sea water is extracted (Hadwiger 1988). from the mass of the collected organism. This How chitosan exerts its effects is not mass is freeze-dried and sent to the NCI for known. It is thought that chitosan has two screening. modes of action. First, fungal walls contain The NCI, in turn, tests each specimen some chitosan. When fungi are stressed by against sixty in vitro cell lines, which repre- unfavorable environmental conditions, they sent seven cancer sites; blood cells, brain, 40 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES colon, kidney, lung, ovary and skin (Ansley Second, some of the organisms constitut- 1990). Extracts are tested for their cytotoxic ing the biofilm produce metabolic by-prod- activity. Those that indicate significant differ- ucts that corrode metals. Sulfate-reducing ential cytotoxicity are given priority for bacteria, in particular, attack metal surfaces, development. In addition, extracts are tested causing local pitting that can lead to cata- for anti-AIDS activity using a human lympho- strophic failure of equipment (Costerton and blastic cell line infected with the live AIDS Lappin-Scott 1989). Bacterial corrosion is virus. Preliminary results from the first set of exceedingly damaging; a 1.75-centimeter screenings are now being compiled and, thick steel plate may be penetrated in about according to a NCI spokesman, a number of six months. Piers, off-shore drilling struc- extracts are cytotoxic. The isolation, purifica- tures, and Ocean Thermal Energy Conversion tion and identification of the pure secondary devices are especially prone to damage by metabolite from promising extracts are ex- organisms constituting the biofilm. pected to take an additional five to ten years; Presently there are two methods for pre- then pre-clinical research can begin. venting marine organisms from forming a Some pharmaceutical companies operate film on submerged substances. First, a sur- well-equipped laboratories that have multi- face may be coated by a nonstick coating tiered screening and testing programs. These similar to Teflon. The disadvantages of this usually are more extensive than those of the method are that these coatings are expensive NCI, including in addition to screens for anti- in themselves, they are costly to apply, and tumor and anti-viral activities, tests for anti- as they age they crack, exposing new sites for inflammatory, insecticidal and herbicidal marine organisms to colonize. Second, the activities (Cardellina 1986). surface may be protected by an anti-fouling Biofllms and biofouling paint, which consists in part of a heavy metal such as copper. The metal slowly dissolves in When a manmade object is submerged in water, in the process creating a toxic environ- ocean waters, marine bacteria quickly begin ment for marine organisms. The problems to settle on its surface. In fact, they prefer to with anti-fouling paints are that they pollute grow on a surface rather than in the sur- the marine environment, they are fairly rounding water. If the settlement process is expensive, and they pose hazards to the allowed to proceed, the initial film consisting workers who manufacture and apply them. Of mostly of microorganisms soon attracts mi- course, once encrustation has occurred, the cro-algae and, eventually, animals such as most common approach is to scrape it off, barnacles and mussels. Surface colonization which is a laborious and damaging operation. will damage the object via two types of Marine biotechnology research is being actions. First, as an ever increasing number directed at solving the problems related to of organisms become enmeshed in the biofilms and biofouling and, concurrently, at biofilm, a noticeable fouling of the surface securing benefits from the natural biochemi- will occur, leading to encrustation. If the cal processes underlying these phenomena. surface in question belongs to a ship, Research at Agouron Institute, California, for biofouling will increase vessel drag, thereby example, is aimed at clarifying the process of reducing its speed and increasing fuel con- bacterial colonization, particularly identifying sumption. It has been estimated that a 200 the genes that control attachment. Research micron thick encrustation on a ship's hull can results may be used to develop coatings that decrease that ship's speed by 20 percent cannot be sensed by the colonizing bacteria. (Curtin 1985). Without this stimulus, they are unable to Marine biotechnology and Its sub-areas 41 settle. R&D having similar aims, but taking marketed in April 1990 for use in cell and a different approach, is being undertaken at tissue culture, but was later discontinued due the Danish Marine Biotechnology Center. to marketing problems. However, Enzone There various terpenoid compounds or com- (Genex's successor) continues to develop a pounds that contain sulfur have been extract- recombinant blue mussel adhesive and has ed from marine invertebrates and are being reached the animal testing stage with the tested in experimental coatings. Some, it is product. The results have been excellent so claimed, are as effective as tin oxide in far, with the adhesive being biocompatible preventing fouling (Cooksey 1991). Yet other and nontoxic (McGuire 1992). scientists, at the University of Maryland, are A second company, BioPolymers of Mary- concentrating on the second step in land, took the more traditional approach of biofouling-when invertebrates respond to collecting thousands of the blue mussel, then chemicals exuded by colonizing microorgan- extracting the adhesive from a gland in the isms by settling on top of them (Curtin mussel's foot. Approximately 30,000 mussels 1985). It may be possible to interfere with were processed to produce one gram of the the function of the invertebrates' chemical glue, which sold for $90 per milligram (Mor- receptors through the release of harmless gan 1990). At the same time, BioPolymer's substances, making it impossible for these chemists studied the glue's structure and were organisms to identify favorable settling sites. able to generate sufficient information about Research to solve the nettlesome problem it to computer-design biosynthetic glues. An of biofouling can, at the same time, be useful endless number of variations of the basic glue in developing products and processes. In fact, structure can be produced, each tailored for the first industrial chemical to result from a specific application. BioPolymers claimed marine biotechnology is a bioadhesive, that to have satisfied the needs of about 300 is, a glue produced by mussels to anchor customers for laboratory adhesives. However, itself to hard surfaces. The advantage of this BioPolymer's process is now in a legal limbo glue is that it sets strongly underwater, mak- because the company filed for bankruptcy in ing it useful for dental and surgical proce- 1990. dures, as well as for certain scientific work. In addition to the two U.S. firms, Univer- Genex Corporation in Maryland began inves- sal Biologicals Limited (in the United King- tigating this glue a few years ago. Genex dom) markets a protein extract from the mus- took the approach of extracting the adhesive sel Mytilus edulis called Cell Tak. Cell Tak is from the foot of the blue mussel and then a transparent material that stays stable for trying to characterize its chemical structure. several weeks at 4° C. It will readily coat While Genex scientists were unable to com- glass, plastic and metals. Its manufacturer pletely unravel the structure, they generated claims that Cell Tak's biological properties sufficient information about it to enable them makes it a favored material for holding im- to design a synthetic gene that codes for a mobilized cells and tissues, thereby simplify- protein having a structure that is 40 percent ing a number of laboratory techniques, such similar to the mussel's glue. The gene encod- as establishing primary cultures, ing the glue was introduced into the genome immunoassays and microinjection (Cell Tak of an industrial yeast; then the engineered 1990). yeast was grown in fermenters where it Although marine biotechnology research produced comparatively speaking large quan- directed at biofilms and biofouling has been tities of the glue (Morgan 1990). A commer- going on for about sixty-five years, much cial product, called AdheraCell, was first remains to be done. In particular, there is a 42 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES need for research to clarify: (1) the effect that microorganisms, such as bacteria and fungi, different surfaces have on adhesion by vari- to break down complex substances is not ous organisms; (2) the mechanisms of the new. Farmers have composted animal wastes biological activity whereby an organism and crop residues since time immemorial not irreversibly attaches itself to a surface; (3) the only to rid themselves of these substances, biochemical nature of the substances that but to simultaneously produce useful by- organisms produce that act to bind them to products, including methane gas and natural the surface (bioadhesives); and (4) the bio- fertilizer. However, attempts to deliberately chemical and biophysiological properties of employ endogenous microorganisms to clean biofouling organisms (Olson, McCleary and up polluted and waste waters are new endeav- Meeker 1991). In addition, little is known ors that are under intense development (Mar- about the differing adhesives produced by the tello 1991). hundreds or thousands of mollusks other than Bioremediation, and its efficient action, the blue mussel. While most of this research depends on a number of conditions including would be basic research, the lesson from the the characteristics of microorganisms present work of Genex and BioPolymer is that one at the site of the process, the chemical make- does not need complete information about the up of the substance or substances undergoing bioadhesive's chemical structure or the mus- bioremediation, the availability of nutrients sel's genetic control over the bioadhesive's vital to the bioremediating microorganisms, production to develop products that are useful the ambient temperature of the environment to transport and specialty chemical industries. where bioremediation takes place, the amount of oxygen available (which determines if the Bioremediation reaction will be aerobic or anaerobic) for the bioremediation reaction, and whether the This general term refers to the use of mi- reaction is taking place in an open or closed croorganisms to break down pollutants and system. Each of these conditions require a wastes in soil or water to harmless or less brief explanation. toxic end-products. End-products may be either simple inorganic chemicals, such as MICROORGANISM. Generally speaking, water and carbon dioxide, or less toxic com- almost any complex chemical substance that ponents of the starting material. Microorgan- is released by natural or human action into isms bioremediate by feeding directly on the the terrestrial or aquatic environment will organic pollutant, by breaking down the eventually be attacked by microorganisms. To pollutant while they catabolize a primary illustrate, an environmental microbiologist at source of carbon, or by secreting enzymes Louisiana State University has over the last that break down the pollutant (Portier and 10 years identified more than 400 different Ahmed 1988). microorganisms and the chemicals they de- While the emphasis here is on the use of grade (Martello 1991). Most often, the pollut- bacteria for bioremediation, it bears mention- ant that is broken down by a microorganism ing that microalgae and seagrasses may also serves it as a source of food. Thus, petro- be used for this purpose. Further, some R&D leum, which is a mixture of hydrocarbons, is has been done to combine bacteria and micro- a very good source for carbon. Environs that algae for waste water treatment; these tech- have over time been subject to pollution by niques may be adapted for wider use in petroleum become populated by microorgan- bioremediation (Sasson 1988). isms that utilize it as a source of carbon. It is The use by man of naturally occurring no surprise therefore, that locations where Marine biotechnology and Its sub-areas 43 natural seepages of petroleum normally oc- SUBSTRATE. The water containing cur, such as the Persian Gulf, are populated pollutants may be considered as a substrate by immense numbers of microorganisms that for the bioremediation process. The substrate have over thousands of years evolved so they is usually quite complex, containing varied are able to utilize petroleum as their sole or chemical structures and constituent elements. main source of carbon. Although the time- For example, hydrocarbons may have short span is much shorter, similar situations exist or long structures; they may consist of a in the soils surrounding long-standing man- single chain of carbon molecules or have made petroleum-extracting structures or complex branches. They may include heavy processing plants. It can be seen that large metals in their chemical make-up, be chlori- reservoirs of naturally occurring microorgan- nated or brominated, or otherwise contain isms suitable for bioremediation may be toxic elements. Pollutants that consist solely tapped for development and propagation. of carbon and hydrogen atoms and that have More often than not, bioremediation simple structures are usually relatively non- reactions are mixed-culture reactions; that is, toxic and easily decomposed by microor- they involve the simultaneous actions of ganisms. Molecules that are highly branched several types of microorganisms. The constit- are difficult to decompose by microbial uents of mixed-culture reactions vary widely, action; those that contain metals, chlorine and dependent on the location where it is taking bromine are toxic to animals, plants and place, the makeup of the reaction mixture, microorganisms alike. In view of the many and many other factors. Because of their possible substrates and the large number of extreme variability, mixed-culture reactions microbial species and genus that have are extremely difficult to study. Currently the bioremedial capabilities, the problem of best way to study and research them is matching organism to substrate is a consider- through the use of microcosms, which are able one. scaled-down models of environments con- structed in the laboratory. A particular micro- uuTrie the sustrate will cosm may be designed to duplicate as closely uayrovidthe major steanc for hes as possilDe the environment and conditions ofr irmdaigmcrb,ta irbs as poil-spiblsite,say the eniroinen d cdiiaouns o continued health will depend on the availabil- an oil-spill site, say the Prince William Sound ity of other nutrients, such as nitrogen and affected by the Exxon Valdez oil spill. This trace elements. Under ordinary conditions, microcosm would include water from the this will not present a problem since the Sound, rock and sand samples from its shore, number of microbes normally present at a an aliquot of the spilled oil, microorganism locale will be limited by the amount of avail- species collected at the site and samples of able substrate and nutrients. Under the ex- other life growing in the water. The ambient traordinary conditions stemming from, for temperature of the microcosm would match instance, an oil spill, an overabundance of that of the Sound and it would go through an substrate suddenly becomes available, but the identical diurnal light cycle. Once the micro- amounts of nutrients remains constant. There- cosm has been set up, experiments can be fore, large-scale bioremediation cannot be performed on it to clarify what happens if successful unless additional nutrients vital to one or another physical parameter is changed; the massive propagation of the bioremediating if a new microorganism is added or an old microbe are provided. Providing additional one deleted; or if various fertilizers are added nutrients is called fertilization. Since situa- or subtracted. tions will vary, fertilizers will have to be for- 44 MARINEBIOTECHNOLOGYANDDEVELOPING COUNTRIES mulated to fit the particular conditions of industry will include both aerobic and anaero- each bioremediation project. bic phases. For instance, aerobic organisms will be used on the surface, while anaerobic TEMPERATURE. Bioremediation is a com- organisms may be employed to treat petro- plex process involving chemical and fermen- leum constituents that have sunk and are tation reactions. Chemical reactions are often amassing on the sea bed. temperature dependent with the rate of reac- tion speeding up as temperature increases CLOSED VERSUS OPEN SYSTEM. A closed until a limit is reached. For this reason, system reaction is one that takes place within bioremediation tends to work better in warm- a confined space, such as in a test tube, flask, er than colder climates. However, fermenta- or fermenter. An open system reaction is an tion reactions do not always follow this rule, unconfined one, such as one that is allowed and when they do, it is to a limited extent. to proceed in the open air or water. Generally The reason is that microorganisms, which speaking, chemical and fermentation reactions drive fermentation, are the end products of an that take place in closed systems are easier to evolutionary process that has been affected by perform, study and control. It is therefore temperature. Thus microorganisms function understandable that most attempts to use best within the temperature range in which bioremediation for pollution control have they evolved. A petroleum-utilizing microbe been done in closed systems (tanker cargo that has evolved in the temperate waters of holds, for instance). the Persian Gulf would probably not function in the cold waters of Alaska. Nevertheless, in Presently, there are two main approaches general, bioremediation will usually proceed to bioremediation of marine pollution (OTA faster in warmer than colder waters because 1991). The first approach is to seed the its rate depends to some extent on physical polluted area with large quantities of mi- and chemical processes. crobes that have been propagated in a labora- tory or pilot-plant fermenter. The microbes AEROBIC VERSUS ANAEROBIC. Without used in this type of endeavor have most often going into detail, the microorganisms that been collected from local sites affected by the may be used to decontaminate, for example, same or similar polluting agent. If local biota the surface waters polluted by an oil spill proves ineffective, petroleum-utilizing mi- would be quite different than those used to crobes collected from other sites may be used treat municipal wastes. The first would neces- to augment efforts. Alternatively, microbes sitate the use of aerobic microorganisms-that that have had their natural capability for is, organisms living in the air and deriving breaking down various pollutants enhanced most of their energy from metabolic reactions via classical breeding and selection programs where oxygen is of primary importance. The may be employed. main catabolic products of aerobic organisms The second approach is to fertilize the are water and carbon dioxide. Conversely, polluted area with nutrients that may be wastes are usually treated in closed systems lacking or are present in inadequate levels. by a sequence of fermentation reactions This is done when it would appear that the involving both aerobic and anaerobic micro- local biota can expand in numbers sufficiently organisms. Anaerobic organisms derive most for effective bioremediation, if only vital of their energy from hydrogen; they may in nutrients were made available. Until now fact be poisoned by oxygen. The main cata- fertilization has been the main approach bolic end product of anaerobic organisms is toward bioremediation, resulting in several methane. Most attempts at bioremediation by successes. A combination approach is also Marine blotechnology and Its sub-oreas 45 possible where seeding is followed by fertil- operating costs, than alternative techniques ization. commonly used in primary and secondary In the future, bioremediation may be done waste water treatments. It is significantly less by microbes specifically designed for that costly than incineration. In general, bioreme- purpose. Scientists at several institutes are diation requires rather low capital investment, genetically engineering microbial species to has low energy consumption, and remains enhance their bioremedial properties. Two self-sustaining (Portier and Ahmed 1988). approaches predominate. First, metabolic Second, bioremediation is more environmen- pathways are being altered to improve the tally benign than alternative waste water efficiency of pollutant degradation. Second, techniques or techniques used to treat pollut- organisms that already are efficient in break- ed or degraded soil. Bioremediation is thus ing down one pollutant, or one fraction of preferable to landfills, surface impoundment, petroleum, are being endowed with added chemical treatment of toxic dumps, and capabilities, enabling them to break down incineration (Portier and Ahmed 1988). pollutants or petroleum fractions that previ- Third, in some cases of marine pollution ously were outside their catabolic range. In there is no real alternative to bioremediation, fact, the first microorganism granted a U.S. excepting natural processes, because conven- patent (in 1981) was one developed by Dr. tional cleanup techniques, consisting of skim- A. Chakrabarty to simultaneously break down ming oil from the water surface, mopping up several petroleum fractions. The use of genet- oil by hand using paper towels, cleaning sand ically engineered organisms in open systems and stone with high-temperature, pressurized is not feasible at present, however, since water, and employing chemical dispersants some believe that it would pose unknown and surfactants, would be too damaging to risks to the environment and society (bio- fragile ecosystems. safety is discussed above). Related to bioremediation is the use of As is the case of any advanced, complex chemicals produced by living organisms to technology, bioremediation poses difficulties treat oil spills. Two types of agents have been while conferring advantages. In regard to tested in the field-biological dispersants and difficulties, the major limitation of the tech- surfactants. Dispersants act to separate oil nique is that bioremediation reactions are spills into small particles of oil, which then often poisoned by chemicals or substances are easily transported from the surface to the that kill microorganisms. This problem is water column and sea bottom. Dispersants especially acute when treating waste waters enhance bioremediation because dispersed oil from municipal or industrial sources whose is more susceptible to attack by microorgan- composition are not completely known and isms than is massed oil. On the negative side, cannot be predicted. An unscrupulous con- dispersants may worsen the effects of the sumer could, for example, choose to dispose pollutants under treatment because of the of toxic wastes cheaply by dumping them in persistence and toxicity of the settled sub- a municipality's sewer. If those wastes con- stance. Shallow coastal waters in particular tained the heavy metal mercury or an organo- could be severely damaged by dispersants. phosphorus pesticide, bioremediation of the Surfactants reduce the surface tension of effluent taking place downstream would likely an oil-water interphase, allowing the oil to be poisoned. emulsify in the water. Surfactants produced As to potential advantages, three bear by bacteria are nontoxic and biodegradable. mentioning. First, the employment of bio- When tested in the Prince William Sound, a remediation to process waste water is less biological surfactant produced by Pseudomo- costly, in terms of both capital expense and nas aeruginosa was found to increase the rate 46 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES of oil removal from sand and rocks on the and (2) it can be used to select plants with beach (Harvey and others 1990). Another especially desirable characteristics, such as biological surfactant, named Emulsan, has resistance to salinity, climatic extremes, acid been isolated from the marine bacterium soils, toxic minerals and so on. Acinetobacter calcoaceticus. It is on the The main limitation of tissue culture is market and is widely used to clean oil hold- that success in regenerating plants from tissue ing tanks in tankers and other ships. Emulsan culture has only been attained with a few is also being tested in applications for en- terrestrial plant species, so many commercial- hanced oil recovery from oil wells and pollu- ly important cultivars, such as wheat, barley tion control (Weiner 1985). and oats, have not yet been regenerated. Cell culture is similar to tissue culture, but Cell culture goes one step further. Rather than generate plants from callus, the callus tissue is shaken, Both animal and plant cells may be used causing it to shed individual cells. Individual in cell culture systems. Yet, in terrestrial cells are picked and placed in culture flasks, agricultural biotechnology, by far the most where they grow and subdivide much like applications have resulted from plant tissue bacteria. Cultured cells can be used in two and cell culture work. These technologies are ways: to generate whole plants or to keep based on the fact that the cells of many plant them in culture where they secrete natural species are totipotent; that is, each cell consti- products that are normally produced by the tuting a plant is able to generate a whole whole plant. plant. Thus, one cell may be micropropa- Cell lines have been developed that pro- gated; that is, by treating the cell in an appro- duce five times as much of valuable com- priate manner, it is possible to raise an entire pounds per dry weight of plant material as plant from it. The practical implications of does the native plant. Products from terrestri- this capacity is that one plant can be used to al plant cell culture processes range in chemi- produce thousands of plants with identical cal complexity from simple sugars to com- genetic makeup without having to wait for it plex polymers; they include drugs, dyes, to bear fruit and seeds (UNDP 1989). fragrances, flavors and pesticides. Cell cul- The micropropagation of a plant is ture products are typically low volume, high achieved by removing a sample from its root cost substances. Already over fifty natural or shoot (an apical meristem) and cutting it in products are being produced in plant cell many small pieces. Each piece is cultured in culture at yields equal to or greater than a test tube or flask with special media that equivalent crops, including quinine, mor- contains plant hormones (this is called in phine, codeine, cacao, pyrethrum, thaumatin, vitro propagation). Cells in these cultured stevioside, jasmine extract, and digitalis. tissues divide many times to form an amor- When considering algal cell culture, it is phous mass called callus. A callus can be important to recognize the difference between further subdivided; the subdivisions can be this technique and aquaculture. Unlike algal cultured to generate shoots, which in turn can aquaculture, the purpose of which is to grow be matured in a green house until ready to be and harvest a large biomass, algal cell culture planted in the field. In this fashion, one gram systems produce a low volume, high cost of callus tissue can generate 500 or more substance. The cells, which have been de- plants. signed and developed to maximize production There are two great advantages to the of that substance, grow and propagate in a technique of tissue culture: (1) it can be used closed system, usually a fermenter or tank. to rapidly mass propagate a selected plant; The culturist precisely determines the compo- Marine blotechnology and lts sub-areas 47 sition of the culture substrate and sets exactly (USA) has developed a process based on a the conditions of fermentation, which is strain of Chlorella that normally synthesizes closely monitored throughout the process. a relatively large amount of the amino acid The product produced by the cultured cells proline. Proline is an important intermediary undergoes a subsequent downstream process- for the production of pharmaceuticals. Re- ing, consisting of recovery, purification and search done by the company's scientists has packaging. led to the development of a strain that pro- While most cell culture work has focussed duces 30 percent more proline than does the on microalgae, one project involving macro- original. Further, since the proline can be algae is worth noting. As noted before, agars extracted without killing the cell, the same are vital to both diagnostic and research organism may be "milked" again and again laboratories. One scientist has gone as far as (Tucker 1985). to say "...most of the major advances in Other industrial accomplishments include modern biotechnology would not have been the French Association for Research in Solar possible without the availability of the poly- Biology developing microalgae for the pro- saccharides from marine macroalgae."(Renn duction of hydrocarbons and polysaccharides; 1990) Unfortunately, the present production the U.S. Solar Energy Research Institute's level of agar cannot meet world demand. The use of microalgae to produce lipids; and the problems lie with inadequate supplies of agar- accomplishment by scientists at Amoco Re- producing algae and poor quality control of search Center, Indiana, in using genetic the natural product due to seasonal variations engineering techniques to introduce the bacte- and differing production methods. To allevi- rial genes coding for the rare amino acid ate the unsatisfactory situation, the European octopamine into the green alga Chlamydomo- Research Coordinating Agency (EUREKA) nas (Tucker 1985). has agreed to underwrite a four year program The production of high priced products costing 4 million ECU (European currency via cell culture can be profitable. Commer- units), or approximately $4.8 million, to cially available products from algal cell develop a cell culture production method to culture include isotopically labeled amino produce agar and agarose. Specifically, cells acids and other compounds (up to $1,000 per from red algae that produce agar will be kilogram), medical phycobiliproteins isolated, characterized and then appropriately ($10,000 per kilogram), food coloring phyco- genetically engineered to increase production biliproteins ($100 per kilogram), beta caro- and to grow well in a bioreactor. This pro- tene ($300-$500 per kilogram), and various gram will be an international effort, combin- amino acids ($5-$100 per kilogram) (Calle- ing the resources of the British PBL compa- gari 1989). An industrialist specializing in ny, the French Pronatec company, and the algal biotechnology estimates that one 130- French Glucide Development Centre. In liter tank can produce $120,000 worth of addition, Pronatec will utilize the molecular specialty proteins and sugars per month biology expertise and resources of the Uni- (Masters 1990). Industry also uses microalgae versity of Lille and the Institute of Technolo- as the active ingredient in immobilized cell gy, Amiens University. Preliminary develop- systems, producing via continuous culture ment and pilot plant trials will be conducted hydrogen, acetic acid, dihydroacetone, and in Ireland and Spain (Polysaccharides 1991). gluconic acid, all valuable industrial chemi- A few cell culture systems based on cals. In addition, microalgae can be used to microalgae are already being used by industry treat wastewater inexpensively. for the production of specialty chemicals. For Very little research has been done on example, the Ethyl Corporation in Louisiana marine animal cell culture. As stated in a 48 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES recent review, "Tissues of fishes are amena- complexity from temperature gauges and pH ble to the techniques of modem cell meters to radioimmunoassay and gas chroma- culture ... and yet this vast resource, compris- tography. However, a new type of sensor has ing of thousands of vertebrate species, re- recently been developed, namely the biosen- mains largely unexplored."(Hightower and sor. A typical biosensor consists of an immo- Renfro 1988) The reason why marine animal bilized biological material-such as an en- cell culture development is behind marine zyme, antibody, or a whole cell-in contact plant cell culture is that it is more difficult with a transducer or signal-generating ele- work. Specifically, animal cells do not form ment, which is a device that converts the callus, the culture media for growing animal information received from the biological cells is more complex and expensive than material into some sort of signal, usually an plant cell culture media, and the culture electric current. Data processing equipment conditions are more exacting for animal cells gauge the reaction by quantifying the signal, than plant cells. Yet, fish cell culture offers providing results based on the data received researchers unique, useful tools for exploring within seconds or minutes. epithelial ion transport, endocrinological Two types of biosensors hold particular studies, response of physiological systems to interest for marine biotechnology. The first is stress, tolerance of organisms to heat and the chemoreceptor, the heart of which con- cold, cancer biology and environmental sists of biomolecular assemblies involved in toxicology (Rechnitz 1988). In view of its physiological functions, such as smell and possibilities for research and applications, it taste, and in metabolic and neural biochemi- will not be too long before fish and other cal pathways. One type of chemoreceptor marine animal cell culture will be common sensor of potential value utilizes the sensing investigatory tools in the laboratory and will antennule removed from the crab. The crab be used in development and production sys- uses this organ to continually monitor water tems for vaccines and other pharmacological for dissolved substances ranging in chemical substances. complexity from simple salts to pheromones (hormones that attract the opposite sex). In Sensors the laboratory, antennules dissected from crabs and connected to potentiometers have Sensors are devices that detect a specific exhibited instantaneous quantitative responses substance or organism. The fact of detection to various amino acids, hormones, nucleo- is made known by the generation of an elec- tides, drugs and toxins (Rechnitz 1988). tric signal, the production of a unique and The second type is the immunological measurable chemical substance, or by other sensor. A monoclonal or polyclonal antibody means clear to the operator. Sensors also are (see page 7) or a DNA probe is the molecular analytical instruments that react to change, recognition element in this biosensor. Modem whether it is a change in temperature, chemi- detection kits based on one or another mono- cal composition of reactants or substrate, or clonal or polyclonal antibody have been another parameter. Sensors may have a dual developed to detect many types of pathogenic function; for example, a sensor initially may bacteria, including those causing cholera, detect the presence of a substance and then shigellosis and typhoid fever. These kits are sense changes in that substance's concentra- highly specific, accurate, easy to use and give tion or amount as a reaction proceeds. results in a matter of four to twelve hours. Conventional sensors commonly employed The second product, the DNA probe, is in laboratories and industry may range in the most exact and definitive way of identify- Marno blotechnology and fts sub-reas 49 ing the type, or even the strain, of virus, 1990). Red tides are so called because peri- bacteria, or parasite that causes disease in odically dinoflagellates will bloom (prolifer- animals and plants. This kind of identification ate in large numbers), giving a reddish sheen depends on a "reverse engineering' feat; a to the ocean water where they grow. Since specific sequence of the genome of virus, many strains of dinoflagellates produce dead- bacteria, or parasite is determined, then a ly toxins, algae (as well as marine animals complementary sequence is synthesized. This such as bivalves) cannot be harvested from sequence in effect becomes the probe for waters affected by red tide. Damage to aqua- testing analytes of unknown composition. culture can run into the billions of yens. This probe, which is labelled with radioactiv- The project has two aspects. First, previ- ity or with biotin, attaches itself to the specif- ous research has demonstrated the existence ic sequence that gave rise to it. The fact of of some marine bacterial species that promote attachment is detected through a procedure the bloom of dinoflagellates and others that that relies on chemical or radiological reac- retard it. The intent is to identify each of the tions. A battery of such diagnostic probes can various retarding microbial species and deter- be put together in a kit, enabling the operator mine which species retards which dinoflagel- to, for example, rapidly identify prevalent late. Next, researchers will attempt to recov- disease agents that inhabit the waters proxi- er, identify and characterize the substance, or mate to a city or region. As is the case with substances, these organisms secrete that monoclonal antibodies, test results can be retard dinoflagellate growth and propagation. generated in a matter of four to twelve hours. Once this has been done, scientists will try to 'Te major advantage of biosensors over mass produce this substance by appropriately conventional sensors are that they are easier genetically engineered bacteria. The hope is to use in some specific applications, give that the inhibiting substance will prove suit- results quickly, can be more sensitive, and able for spreading over an area affected by are more selective. For example, in industrial the red tide. bioprocess control biosensors are used for Second, researchers will seek to identify real time monitoring of complex reactions, each of the dinoflagellate strains that consti- giving technicians and engineers timely warn- tute the red tide. Monoclonal antibodies will ing to adjust reaction rates to achieve maxi- thereupon be constructed for each strain. mum production. As the technology advanc- Once this has been accomplished, waters es, rugged, inexpensive, accurate biosensors threatened or affected by red tide can be will be developed for field use, possibly to sampled and the causative dinoflagellates be the point where they will be disposable after identified quickly. Timely treatment can then being used a single time. A large variety of be instituted with the correct inhibiting sub- biosensors will be used for a plethora of stance. purposes in biotechnology industry, food The Japanese project has important impli- production, agricultural projects, environmen- cations for developing countries because tal studies, and medicine and veterinary many are affected by red tides. For example, medicine. the Chilean Under-Minister of Fisheries, Mr. An example of a project in which bio- Andres Couve has claimed that the red tide is sensors will play a vital role is one under more of a threat to his country than cholera. way in Japan. In 1990, the Japanese Fisheries He notes that in 1991, 41 Chileans became Agency launched a five year biotechnology- sick with cholera, but over 300 persons were based project to prevent damage to macro- poisoned by red tide (Couve 1992). The latter algae aquaculture from red tides (Five-year number is probably an underestimation since 50 MARINEBIOTECHNOLOGYANDDEVELOPING COUNTRIES many who consumed contaminated mollusks As was noted in Chapter 1, biological did not report their sickness or were misdiag- oceanography, while extremely important, is nosed. a field that has been relatively neglected due The marine environment is highly variable to limitations inherent to present-day detec- and exceedingly complex. The inter-relation- tion and measurement techniques. Biotechnol- ships in the oceans between biological activity ogy techniques that are now used on land can and physical and chemical processes are be adapted or further developed to study largely unknown. Yet they must be under- problems in biological oceanography. As has stood before events of global ecological been described in a recent report: significance can be reliably predicted and their effects measured. While much of the [Biotechnology] makes possible the rapid ocean surface and the overlying atmosphere identification and quantification of species, is continuously monitored by sensors em- stocks and populations of marine organ- placed on anchored and free-floating buoys, isms from microscopic viruses and bacte- on-site sampling, or remote sensing methods, ria to the largest marine mammal; identifi- biological and chemical activity in the water cation of the mechanisms regulating the itself is difficult to detect and measure by reproduction, development, evolution, present technologies. As a result, there is a growth and distribution of marine organ- dearth of data on inorganic and organic isms; identification of the mechanisms pollutants that are present in ocean water (and controlling the interaction of marine or- often have poor water solubility and are ganisms with one another and with their therefore present in low concentrations) and environment; and manipulation of these on intact microorganisms that inhabit waters regulatory mechanisms for enhanced on a temporary or permanent basis. Clearly, production of valuable marine resources, many specific sensors need to be developed for bioremediation of environmental in- to detect and measure all the varied biological sults, and possibly even for the bioremedi- and chemical processes in the marine envi- ation of changes in greenhouse gas accu- ronment so they can be adequately studied. mulation and the rate and extent of clima- tic change (Initiative 1990). Biological oceanography (including public health) The report delineates three areas in bio- logical oceanography that can benefit from Biological oceanography is the study of the application of biotechnology. First, bio- "the ecological relationships of organ- technology techniques can be used to increase isms-their interactions with other organisms our understanding of how the distribution, and with the features of their environment, abundance and growth rates of marine popu- particularly larger scale features such as lations are regulated. Techniques such as currents, bottom topography, and water mass restriction fragment length polymorphism characteristics" (NRC 1985). The interrela- (RFLP), DNA/DNA hybridization and poly- tionships between the environment and its merase chain reaction (PCR) will permit microbial inhabitants is the concern of a scientists to characterize the genetic makeup scientific field called microbial ecology of populations, to perform comparative analy- (Updegraff 1991). The interactions between sis of different populations, and to discern microbial ecology and humans fall within the patterns in genetic variation within species. purview of a new scientific specialty called The practical applications from increased environmental toxicology. knowledge will be used to identify and track Ma,lne blotechnology and ts sub-aresa 51 larval forms of invertebrates; identify the microbial primary producers, including country of origin of commercially important microplankton (which have recently been fish stocks, such as salmon; develop plasmids proven to be responsible for most of the in macroalgae as vectors; and to characterize production in the oceans). This research is of marine microbial populations. The last listed interest because it clarifies one aspect of is particularly important since researchers are impact of microbial activity in the oceans and able to culture only about 1 percent of marine the regulation of gas exchange between air bacterial species. and sea. This in turn bears on the interrela- Second, biotechnologytechniques, particu- tionship of the cycles of nitrogen, phospho- larly rDNA and PCR, may be employed to rous, oxygen and others that interact with the research how marine organisms adapt to their carbon cycle; which is a vital determinant of varied environs, clarify interactions between global climatic change and the accumulation species, map the evolution of species, and of greenhouse gases in the biosphere. explain the symbiotic relationships between Environmental toxicology has particular organisms. There are many possibilities for importance in matters related to the coastal applications from this research. Some have zone, because this is where land meets sea, already been discussed; for example, by where most of the world's human population clarifying the genetic control over the produc- lives, and whose terrestrial and marine envi- tion of enzymes by extremophiles, new possi- rons teem with microorganisms. To illustrate, bilities open up to manufacture unique en- research done at Woods Hole Oceanographic zymes useful to industry. Other possibilities Institution in Massachusetts has shown that for terrestrial agriculture are discussed in the more than one million bacteria can inhabit next section. In addition, gene probes can be one milliliter of water and almost one billion used to identify intermediate life stages of bacteria can be found in one gram of beach important organisms that now are unknown, sand. such as shellfish; the chemical signals that In addition to the microbial populations regulate settling and metamorphosis in shell- "naturally" inhabiting beaches and coastal fish can be identified and used in aquaculture; waters, microorganisms are continuously and the genetic control over the production of introduced to these settings through human adhesives by mollusks can be elucidated, activities. It is unquestioned that huge quanti- opening up possibilities for new products. ties of raw sewage and other waste is dumped Third, biotechnology techniques may be untreated in the oceans every day. Some of employed to increase our understanding of the consequences of this massive pollution the factors that regulate interactions between may become discernable rapidly, as has been organisms and the environment. Specifically, shown in Chile. There the uncontrolled research can be aimed to research primary discharge of large amounts of untreated productivity, which is the all-important pro- wastes along Chile's 4,200-kilometer long cess whereby inorganic carbon is taken up by coastline has led to many cases among swim- organisms (mostly bacteria and plants) and mers of hepatitis A and E, as well as a few converted by them using energy from the sun hundred cases of cholera (Rojas 1991). In into organic carbon, which in turn is the basis addition, banks of mollusks (clams, mussels, for all forms of life. Sophisticated instru- oysters and piures) are located just off shore ments and techniques used in biotechnology where waste water is dumped. Many Chileans research, such as bio-optics, flow cytometry, have become sick after eating them. immunofluorescence probes and biological However, in most instances the ultimate markers, can be used to identify and quantify effects of wastes dumped in the oceans are 52 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES unknown. We know little, for example, about was granted a license by the U.S. Food and the fate of the many types of pathogens Drug Administration to market the product. released in the marine environment, the The major application of LAL today is in the interactions between bacterial pathogens and testing of injectable pharmaceutical products marine viruses, and the probability of genes for endotoxins, which it can detect at levels from bacterial pathogens dispersing widely. lower than the pyrogenic level. Its use as a One can theorize that persons who are ex- diagnostic tool is limited, however, because posed to large numbers of bacterial and viral it does not differentiate between the many pathogens inhabiting waters directly (for types of endotoxins that may be secreted by example, while swimming or washing) or bacteria (Novitsky 1984). indirectly (as a result of, for example, eating To conclude this section, two subjects that marine animals that have been harvested from are related to possible marine biotechnology contaminated or polluted waters) will suffer applications for public health are noted. The increased rates of morbidity and mortality, first is that marine animals offer themselves but hard data on the matter is lacking. The as models in biomedical research. For exam- major problem is the difficulty investigators ple, the icefish, which lacks red blood and have with collecting and identifying patho- hemoglobin, may be used to study anemia; gens in the oceans, then establishing cause- the immunoprotective mechanisms of sharks, effect relationships between pathogens and which rarely suffer from cancer, may be used health events. The development and market- to develop cancer treatments; the shark also ing of detectors (see above) and precise, has an extremely high blood level of urea that easy-to-use diagnostic kits based on mono- may be used to study uremia in humans; and clonal antibodies and DNA probes will help the sea cucumber, whose peritoneum (abdom- investigators perform the basic research that inal cavity) is filled with bacteria, is able to would clarify the effects that contaminated protect itself from peritonitis (Smith 1988). and polluted sea water have on public health. Second, as is well known, seafoods are Shifting our focus from environmental consumed throughout the world, providing toxicology to marine products of public high quality protein and other important health significance, one of the earliest appli- dietary constituents. However, seafoods can cations from "classical" marine biotechnology be the source of foodborne diseases due to is related to the detection of toxins important infectious organisms or toxins. Seafoods have to public health. It stems from a 1955 discov- been implicated as the vehicles for 10 percent ery that the blood from the horseshoe crab, of all foodborne diseases in the United States, Limuluspolyphemus, would clot when mixed 7.6 percent in Canada, and 13.2 percent in with endotoxin produced by certain bacteria the Netherlands (Huss 1991). There is intense called gram-negative bacteria (so named interest in industry to develop biosensor kits because they color red when stained accord- designed to detect low numbers or levels of ing to a process developed by C. Gram). A specific pathogens and toxins in seafood. Kits purified extract of the blood suitable for based on monoclonal or polyclonal antibodies diagnostic work was prepared in 1964 and seem to hold particular promise in the short was named Limulus amebocyte lysate (LAL). term. As kits are marketed and used routinely After the large pharmaceutical companies in the seafood industry, contaminated batches showed their disinterest in LAL, its developer of seafood will be detected early and prevent- started his own company to commercialize it. ed from reaching consumers. A marked Thus, in 1974, the company Associates of reduction in the incidence of foodborne Cape Cod, Inc., was formed, and in 1977 it diseases will undoubtedly follow, saving Marlno botochnology and Its sub-reas 53 millions of dollars in health costs and pre- to marine animals and plants would benefit venting much misery. their terrestrial counterparts. Some researchers are trying to answer this Terrestrial agriculture question. For instance, a team at Canada's Plant Biotechnology Institute, Saskatchewan, Cyanobacter and other microalgae, which is trying to improve the cold hardiness of are normally present in wet soils, possess the crops subject to damage from frosts that may valuable attributes of nitrogen fixation, ability unexpectedly occur in the early spring and to stabilize soil aggregates, and the ability to late fall (Cutler, Saleem and Georges 1989). produce plant growth regulators. Thus it is no The team has taken a three-phase approach in surprise that these aquatic organisms, espe- tackling the project. First, its members col- cially the cyanobacter Azolla, have been used lected the anti-freeze protein from flounder. for a long time by farmers in India and The protein was used to infiltrate different southeast Asia as green fertilizer. More plants, which were subjected to freezing. The recently, mass culture technology has been freezing point of leaves of treated plants was adapted in India to produce free-living cyano- found to be 1.8° C lower than untreated con- bacteria for use as biofertilizer and soil con- trols. Second, laboratory findings were evalu- ditioner. The process is based on starter ated in terms of practical field applications. cultures produced by the Indian Agricultural Based on meteorological data collected over Research Institute; the fertilizer itself is five years in Saskatchewan, the lowering of distributed as dried flakes. Cost analysis of the freezing point of crop plants from oa to cyanobacter biofertilizer use versus chemical -2° C would decrease the number of dam- fertilizer shows conclusively the superiority aging frosts from 22 to 5. The third phase, of the first (Metting 1989). While the devel- which is a proposed one, involves introducing opment of microalgae as biofertilizer (also the gene coding for the anti-freeze protein called algalization) has so far been done by into an appropriate crop plant. The research- researchers using traditional research meth- ers involved in the project believe they can ods, future research on strain selection and use the bacterium Agrobacterium tumefaciens improvement could be significantly advanced as a vector for introducing the gene into the through the employment of advanced biotech- target plan. However, a key problem remains nology techniques. to be solved by the Canadian group, namely, Compared to terrestrial life, marine organ- to clarify the regulatory sequence that con- isms possess characteristics that in some cases trols the expression of the anti-freeze gene. are unique, in others merely unusual. For The natural gene is controlled by a sequence example, the world's most salt-tolerant plant that works only in the fish, so finding one appears to be a certain genus of Dunaliella that works in the plant will be the next step. that inhabits the Dead Sea, which contains 29 A similar problem may have been solved percent salt (Weiner 1985). The fastest grow- by the U.S. company DNA Plant Technology ing plant may well be the giant kelp, which Corporation in New Jersey. Company offi- has an estimated maximum growth rate of 65 cials claim to have synthesized the flounder centimeters per day (Robinson 1985). And, anti-freeze gene and have inserted it in yeast as mentioned, the winter flounder survives a and higher plants, where it was expressed. sub-zero temperatures that would kill most The transformed yeast's viability after freez- animals, including other fish. One cannot but ing was increased by 200 to 300 percent, a wonder, if it were possible to do so, whether quality which will be useful for bakeries that the transfer of certain characteristics inherent produce frozen dough products. In regard to 54 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES higher plants, the company is asking the dryness are recorded. The results were ex- USDA for permission to field test a trans- tremely promising; the halophyte Salicornia genic tomato containing the gene. It is hoped bigelovii was found to be "...a potentially that fruits and vegetables containing the gene valuable new high-yielding oilseed crop for will retain most of their fresh character after these regions, yielding a vegetable oil high in freezing and thawing (DNAP plants 1991). unsaturated fatty acids, which is amenable to These findings have implications for other commercial oilseed extraction methods" parts of the world. It is estimated that, for (Glenn and others 1991). example, California and Florida citrus fruit This emerging area in agriculture is of agriculture, which is sometimes devastated by special importance to developing countries unseasonal frosts, would save millions of because arable lands are fully utilized in most dollars if the citrus trees were able to survive of these countries, particularly those that are 2° C colder temperatures than they can now. densely populated, while, concurrently, huge It does not take much imagination to realize tracts of what are termed marginal lands are that large areas in developing countries, such not used, or are underutilized, because for as Afghanistan, Chile, Kenya, Nepal and reasons of economy or unavailability they Peru, would benefit greatly if crops planted cannot be irrigated by fresh water. In addi- in soils located at high altitudes could be tion, unsound irrigation practices have led to protected from frost. the degrading of tracts of land due to the The scientific or technical area where salinization of soil. Modern biotechnology marine biotechnology immediately intersects now makes it possible to utilize the so-called with terrestrial agriculture is saline agricul- "marginal' lands by providing researchers ture. In fact, a tremendous amount of re- with the tools to develop crop plants that search is being directed at developing plants thrive when irrigated by brackish or salt that are salt-tolerant or require salt water for water. Once this development has occurred, life (halophytes). The significant advances the concept of marginal lands will change; that have been achieved in these areas of henceforth the problem will be one of match- research have been reviewed by the NAS ing a land having certain chemical and physi- (BOSTID 1990). More recently, a terrestrial cal characteristics with a crop that is best halophyte was evaluated after six years of suited to grow where these characteristics are field trials in the Sonora desert, Mexico, present. where conditions of temperature extremes and 4 Options in marine biotechnology for developing countries In view of the limited resources available this fact, we nevertheless hold that countries to most developing countries, R&D likely to whose bioscientific capability is low could produce applications in the short term (one to begin working in marine biotechnology (or three years) and medium term (three to six biotechnology-related) R&D somewhere years) would probably have higher priority along the gradient, depending on resident than long-term projects. Further, two impor- expertise and available resources. Specific tant factors must be taken into account when examples that support this contention are considering options in marine biotechnology provided in the next section. for a developing country: (1) the level of When moving from the scientific under- scientific or technical capability in the coun- pinnings of marine biotechnology to its more try in biology, bioscience and biotechnology, applied aspects, many developing countries as well as in applied areas, such as aquacul- possessing or controlling marine tracts have ture and fisheries; and (2) the potential of established significant marine-related indus- prospective R&D resulting in applications for tries, usually aquaculture or fisheries. These that country in the short to medium term. industries may range in complexity from The two factors need to be considered in simple (for example, a shrimp pond operated detail. by a family) to complex (for example, a facility consisting of pools for growing algae, Existing scientific and fish or shrimp, a pumping system for circu- technical capabilities lating fresh salt water, and a processing plant). Some of the larger, more diversified As is made clear in Appendix A, research operations may include a research unit, institutes in the fields of marine biology and staffed by a few technicians, who are respon- marine biotechnology-related areas are scat- sible for solving immediate problems pertain- tered throughout the developing world. Un- ing to water quality control, diseases affecting doubtedly, their capabilities vary widely; for cultured animals and plants, feed and nutri- example, many have the capability to perform tion, and waste management. rudimentary experiments in marine biology, For countries that possess a marine indus- while a few could take on complex projects try beyond the "family farm stage" and that such as studying and developing transgenic wish to develop capabilities in marine bio- fish. Most of these research institutes would technology, two options exist. First, aquacul- thus fall at the lower end of the capability ture enterprises that have research units can gradient depicted in Figure 3. The progres- be encouraged to expand and upgrade by sion from capability to perform simple exper- hiring doctoral-level natural scientists and iments in biology to being able to clone genes purchasing the scientific equipment required is not accomplished easily; the capability to perform applied R&D. The home govern- required for R&D involving transgenic organ- ment may stimulate such development by isms is on an order of one or two magnitudes granting appropriate subsidies, allowing firms more demanding in terms of expertise and tax credits, and instituting other incentives. equipment than are "classical" investigations Second, universities and research institutes in biology and bioscience. While recognizing involved in marine matters can set up out- Figure 3: Gradient of Marine Biotechnolgy *Advanced' Technologies Development of Transgenlc Rsh ' FINFISH Characterizabon of Selectve Genes 7 Development of FINFISH (e.g., Growth Rate) Exprssio System - Cloning of Genes that Encode Development of Triploid Fish - Bioacitve Compounds .- / Structural Elucidabon of Metabolites Optmizabon of Aquaculture " Optmization of Production by Production System 2 Fermentation of Metabolites Isolabon of Specific Metabolites Selective Breeding - , / ~~~~~~~~~MARINE BACTERIA Isolabon and Identfication of Marine Bacteria that Produce Bloactive Compounds Screening for Bioactive Compounds *Classical- Aquaculture , Technologies X Fermentation Increasing Manpower and Equipment Demands EKhv53292a Options In marlne blotechnology for developing countries 57 reach programs through which their research Singh 1991; Yap 1990). In addition, aqua- units establish working relations with indus- culture may be undertaken to generate new try. The underlying basis for cooperation applications including, for example, to pro- would be that both sides benefit. The industry duce marine leather from large fish such as could, for instance, provide sites for field seabass (barramundi) (Selwood 1992), salt research, while the research institution helps water pearls (McElroy 1990) and fresh water industry by performing problem-solving pearls (Pearls 1992). The expansion of aqua- research or research aimed at enhancing culture, however, will take place through the production. Implementing either option, or a employment of mostly well-known traditional mix of the two, would help countries gain technologies, amplified at times by tissue their goal. culture. Some countries may be fortunate in that What is the future role of advanced bio- their scientific institutes already have signifi- technology in aquaculture in the developing cant capabilities in biotechnology and their countries? One role, of course, is in the marine industry has applied skills in aquacul- control of animal diseases, which is dealt ture or other areas. In these cases, the major with in the next section. Another is to devel- emphasis of government initiatives might be op triploid and transgenic fish or shellfish to bridge the gap between the research estab- specifically for aquaculture, for the purpose lishment and the applied sector by fostering of increasing yields or to produce a superior joint projects between them (Zilinskas 1989). product: one that is more attractive, better tasting and more nutritious than present Applications of marine biotechnology in stock. Is this proposal realistic? the short and medium terms Many institutions in developing countries have, or could shortly develop, the R&D Based on the history of terrestrial biotech- capability to produce triploid oysters and nology development and assuming that ma- other mollusks. This could be a worthwhile rine biotechnology will undergo a similar endeavor because industrial countries whose growth and maturation process, it is possible populations relish mollusks are facing de- to estimate when applications are likely to creased production due to the shrinkage of emerge from R&D in each of the nine sub- areas where they may be cultured, increased areas of marine biotechnology we listed in pollution that kills off the animals through Chapter 3 (aquaculture, marine animal health, disease, and high labor costs. This opens up marine natural products, biofilm or bioad- new possibilities for Asian and Pacific coun- hesion, bioremediation, cell culture, biosens- tries to culture mollusks and export them to ors, public health, and terrestrial agriculture). receptive markets in Europe, Japan and North America (Newkirk 1991). A multi-year Aquaculture attempt to do so is in fact under way now in Western Samoa. About three years ago, the In general, aquaculture presents many South Pacific Aquaculture Developments opportunities for developing countries in Project, based in Fiji, imported diploid and traditional areas; to increase their food sup- triploid oysters, which were cultured locally. plies, improve the nutritional status of their Of the first batch, all the diploid forms died populations and generate export earnings. after spawning, but 10 percent of the triploids Countries in Asia and Latin America have were harvested and sold at a premium price. seized on these opportunities and have as a The commercial viability of the project is result benefitted in real economic terms now being assessed by the Western Samoan (Csavas 1989; Ferdouse 1990; Gotfrit 1990; government (More 1992). The findings would 58 MARINE5IOTECHNOLOGYAND DEVELOPING COUNTRIES have implications for many countries in the standards that have to be met by products not region that control large ocean tracts and do intended for human consumption, securing a not have to contend with polluted waters. If permit to perform the testing of a transgenic it is deemed economically viable to culture macro-algae in a closed system should not mollusks in the South Pacific, it would proba- prove difficult to procure. But a proposed bly be advantageous to develop an indigenous transplantation of the transgenic macro-algal R&D capability to produce triploids, thereby species to open waters would be likely to foregoing the need to import them. raise difficult issues, as discussed on pages Although transgenic fish have been devel- 18. Environmental impact assessments would oped with favorable growth characteristics, have to convincingly demonstrate that the they are not likely to be cultured on a large transgenic algal strain would pose no hazard scale until questions about their safety as to the environment, something that would human food have been settled and the risks take years to do. We conclude that until an they may pose to the environment if acciden- adequate regulatory regime is in place, which tally released assessed. The food safety issue is likely to take at least five years, no one can be settled through testing carried out on should culture transgenic macro-algae in open animals raised in the laboratory. But before systems. The propagation of transgenic mac- field testing can commence, the environmen- ro-algae in a closed system would probably tal impact of the possible release of the trans- not be economically feasible. (The culture of genic fish has to be assessed. The results micro-algae is discussed below, in the section from the assessment would be the basis for on cell culture.) regulations governing field testing, which would have to be formulated and implement- Marine animal health ed by the appropriate national executive and legislative bodies. The undertaking of appro- Fish, crustacean and mollusc aquaculture priate safety testing of transgenic animals and is important to many developing countries, the development of a regulatory regime are both as a source of high quality protein and likely to take a long time. Therefore, signifi- to generate export earnings. A wide variety cant applications from the development of of marine species are being cultured, for transgenic fish, shellfish or mollusks cannot example, in Asia fifty-two finfish, sixty-three be expected in the short or medium term. mollusc, eighteen crustacean, and forty sea- Many types of terrestrial transgenic plants weed species are culture in one or more have been developed and have been, or are countries of the region (Csavas 1989). Con- being, field tested. Several industrial coun- sidering the damage that infectious diseases tries have formulated regulations governing have caused to the aquaculture industry, these tests; these regulations would most deploying marine biotechnology for R&D probably apply to the field testing of trans- leading to vaccines against diseases common genic macro-algae in closed systems. Al- in aquaculture seems worthwhile. Some R&D though as far as we know no transgenic is being expended for this purpose. As noted, macro-algae has been sufficiently developed several vaccines developed via conventional for field testing, such an advance is likely not means are available to protect fish and lobster too far off in time. For instance, a first test from infectious diseases, and a recombinant could involve a transgenic macro-algal strain killed type vaccine against IHN has been that has been developed to overproduce agar. developed and is close to field testing. How- In view of the more relaxed rules governing ever, in view of the diversity of marine life the testing of transgenic plants in, for exam- being cultured, which is susceptible to nu- ple, greenhouses, and the less stringent safety merous diseases, and considering the present Options In mardne biotechnology for developing countries 59 level of effort, much remains to be done in usage in aquaculture would decline, reducing the animal health area. risks to public health and environment. The large scale field testing of several Shrimp aquaculture generates significant genetically engineered terrestrial animal export earnings for several Asian and Latin vaccines, including vaccines against rabies American countries. To illustrate, worldwide and foot-and-mouth disease, is now taking aquaculture production of shrimp reached an place in several nations. Considering that the estimated record 690,100 tons in 1991; the regulations for field testing animal vaccines largest producers were China (145,000 tons), are in place, permission to test a recombinant Indonesia (140,000 tons), Thailand (110,000 killed type fish vaccine should not prove tons), Ecuador (100,000 tons) and India difficult to secure in the United States The (35,000 tons) (Record year 1992). Infectious development of similar vaccines is taking shrimp diseases have spread among these place in other countries, including England countries, causing enormous economic dam- and Norway. Field testing this type of vac- age (see page 32 and following). Yet, no cine in England will probably not present a vaccines are available to prevent shrimp problem, but many Norwegian citizens ap- diseases. The reason is that little is known pear to be suspicious about advanced biotech- about the immunological defense systems of nology, so public opinion may prevent field most crustacean species. The basic research testing in that country. that could provide this information is lagging, It is probable that R&D is under way in mostly because these problems have little industrialized countries on recombinant atten- relevance to researchers in the industrial uated type vaccines. For example, a vaccine countries. Therefore, this is one research area may be based on a disease organism that has in which scientists in the developing countries been attenuated by researchers removing from where shrimp aquaculture is important could its genome genes that code for virulence. excel. However, this research would be This organism would presumably elicit an mostly basic research, so applications should antibody response in the host without causing not be expected until the medium or long disease. However, since the vaccine is consti- term. tuted by living organisms, its proposed test- Little is known about the immunological ing would present problems since the vac- defense systems of mollusks. How to best cine's safety must be proved. protect these animals from diseases is there- From a technical viewpoint, R&D in some fore still an open question. However, the of the more advanced developing countries, major constraints to mollusc aquaculture does such as China, India, Thailand and others not seem to be disease, but manmade pollu- could be directed at producing recombinant tion of the mollusc natural environment and vaccines against fish diseases, and results the occurrence of red tides and other algal would probably be realized in the short term. blooms, which render the animal inedible due If the vaccine is a killed type, its testing to it accumulating toxins (Lovatelli 1990). would probably be permitted in most of these These problems fall under the purview of countries as long as established testing proto- biological oceanography, below. cols were followed and local laws were adhered to. If the new vaccines proved effica- Marine natural products cious, local aquaculture industry would bene- fit because the prevention of disease would Several wide-ranging, large-scale projects increase yields of treated animals. In addi- are under way in the tropical and semitropical tion, if vaccines were developed that protect- seas to collect marine animals, plants and ed animals from bacterial diseases, antibiotic microorganisms and screen them for natural 60 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES products useful as specialty chemicals or one compound ends up as a marketable prod- pharmaceuticals. The first phase, that of uct. The demands of this development pro- collection, may be relatively easily done if cess are particularly high if the substance of surface and sub-surface organisms are to be interest is intended for human use. For exam- collected, or if the target organisms live in ple, as of this writing, Didemnin B (see page accessible sites, such as mudflats or shallow 36) is still undergoing clinical trials twelve seabeds. This type of collection can be under- years after discovery. If its developers suc- taken by most countries. However, some cessfully conclude initial clinical trials, anoth- collection will be costly because it is difficult er five years or so might pass before they to perform, for example, the collecting and complete advanced clinical trials and the drug screening of extremophiles. Since very few is licensed. The entire research, development countries are by themselves equipped to and testing process could cost $150 million or undertake the collecting of organisms in the more, which must be paid up front before the abyssal environs, this type of activity encour- drug is marketed. The process to develop a ages joint, cooperative efforts between coun- substance for use other than as a pharmaceu- tries wherein resources and expenses are tical or food additive would be much less shared. costly. Thus, the cost to develop a pesticide The comprehensive screening of organ- or animal feed additive may be about isms for valuable natural products is a costly $10-$20 million. and technologically difficult activity. As a Clearly options vary widely in the natural result, investigators tend to design screening products area. Almost all countries possess programs that cover a class or category of the capability to collect organisms from easily activities; that is, antibiotic, antiviral, pestici- accessible sites; a few of these countries are dal or toxin compounds. Examples of such able to screen the collections in some man- programs were described above, including ner. If a developing country wishes its re- that by the NCI to identify possible anti- search institutions to do more than rudimenta- cancer and anti-viral substances. In view of ry investigations, its researchers and laborato- the difficulties attendant to screening, projects ries may consider entering into mutually that seek to collect and screen animals, plants beneficial cooperative agreements with coun- or microorganism should be cooperative terparts in industrialized countries. Interna- projects, involving both public institutes and tional cooperation is particularly important in private companies that already have experi- cases when a substance shows promising ence in this type of endeavor. It bears men- activity against dread diseases, such as cancer tioning here that the screening for natural and AIDS, or illnesses affecting the cardio- products having interesting properties is done vascular system. In these cases risks may be in the same manner whether the organism high (in terms of the substance not fulfilling under investigation is of marine or terrestrial its initial promise), but if testing is success- origin. ful, the payoff probably will be bountiful. Screening is, of course, only the first step However it is done, natural products develop- of a long development process. The experi- ment is a long term effort. ence of pharmaceutical companies indicates There are two possible exceptions to the that out of 10,000 screenings only 20-30 foregoing. The first relates to toxins. Purified compounds will be identified as possessing marine toxins costs thousands of dollars per interesting bioactivity. These compounds are milligram, yet there is an unsatisfied world subject to a long, involved and expensive market for them. Due to the market demand process to develop and test them. Eventually, for these substances, countries possessing Options In marine blotechnology for developing countries 61 tracts in the tropical seas should consider producing 1 kilogram of sponge comes out to making the investment necessary to be able to $3 (Shang 1991). It can be seen that if and collect, screen and process marine organisms when a sponge species is proven to be the rich in toxins. It would be vital to involve source of a commercial product, it probably private industry in such endeavors. The would make economic sense to establish an second is chitin R&D. Chitin is abundant in enterprise that would undertake the large- much of the developing world; sometimes it scale culturing of the sponge, would harvest is a significant local pollutant. Its derivative and clean it, then process it to isolate and chitosan is therefore an inexpensive resource recover the desired natural product. In addi- that, as we have seen, has possible applica- tion, such an operation would be enviromnen- tions in agriculture. For these reasons, re- tally benign because sponge aquaculture is search aimed at clarifying chitosan's pestici- essentially nonpolluting and the availability of dal properties should be encouraged as it cultured sponges would prevent unscrupulous would be likely to lead to important applica- collectors from depleting wild species. tions in the medium term. Locally produced and processed chitosan could replace import- Bioflims and blofouling ed, polluting chemical pesticides. However, large investments in chitin processing and The study of the biofouling process, and chitosan production should not be made until the resulting biofilm, consists mostly of basic the market demand for these substances has research to explain the settling phenomena been determined. and the molecular biology and biochemistry It bears mentioning that there undoubtedly of organism-surface interfacing. Research are possibilities for a nation to take up R&D findings could be applied to protect ship that combines two sub-areas; for example, hull's and marine structures from encrusta- aquaculture and natural products develop- tion, while eliminating marine paints that are ment. To illustrate, more than 5000 species sources of pollutants. However, in view of of sponges exist worldwide, out of which the difficulties inherent to this research and about 15 have commercial value. Sponges considering the scarcity of data in this area, now have limited uses, mainly for pottery biofouling investigations are not likely to lead making, painting, polishing and washing. to applications in the short and medium term. Total world production is 160-270 tons per The one exception to the foregoing is year; Tunisia and Cuba are the main produc- research centering on biological adhesives ing countries (Josupeit 1991). As was men- from marine organisms. As we have men- tioned above, however, sponges are rich tioned, some information exists about mussel sources for interesting natural products, adhesives, including the identification of including some that have been developed to some genes that control for their production the point where they are undergoing clinical and the elucidation of parts of the chemical testing. structure of the adhesive itself. Researchers in In general, sponges are relatively easy to some developing country institutions should cultivate, reaching commercial size in five to be able to build on the already existing scien- seven years. It should be feasible to culture tific base in this area, and add original find- species of sponges that are the source of ings from research on indigenous marine valuable natural products. The experience of organisms. By taking this short cut, applica- Micronesia indicates that it costs $269 over tions in the marine adhesive area could be two years to culture a unit of sponges, where realized in the medium term. We note, how- the unit consists of 1188 sponges. The cost of ever, that if the intended application is for 62 MARINE BIO TECHNOLOGY AND DEVELOPING COUNTRIES human use, for applications in surgery or activity, genetically engineered organisms are dentistry for example, the same consider- not likely to be used for bioremediation in the ations regarding safety and clinical trials that open marine environment for the foreseeable were discussed under pharmaceuticals would future. apply. 7lssue and cell culture Bioremediation Marine plant tissue and cell culture R&D Effective bioremediation technologies, presents a bit of a quandary. On the one which most often depend on the actions of hand, in many developing countries capabili- naturally occurring microorganisms, have ties exist that would allow their researchers to been developed and their methodology has commence relatively quickly and easily pro- been published. Developing country scientists jects to improve marine plants through tissue who have a background in areas such as and cell culture. This situation stems from waste water treatment and anaerobic fermen- researchers having employed terrestrial plant tation could be trained in bioremediation tissue culture in these countries for a decade technologies in six to twelve months. If or more, resulting in a significant reservoir of appropriately trained scientists are available, knowleJge and know-how having been built they could quickly turn their skills to devel- up. Further, some of the techniques devel- oping and adapting existing bioremediation oped for use in terrestrial plant biotechnology technologies for indigenous use. The actual may be use in algal R&D. These techniques application of bioremediation to clean up have been extensively described elsewhere already polluted coasts, or to alleviate the (UNDP 1989; NRC 1990). However, the effects of future pollution could commence in important point about plant biotechnology, the short term. Once experts have been and its possible adaption for marine plant trained and are working in well-equipped R&D, is that the levels of difficulty inherent facilities, original R&D to, for example, to applying them vary greatly. This means recover, investigate and evaluate new groups that most laboratories will be able to enter of pollutant-destroying microorganisms could into algae research for species improvement, also commence in the short tern; applications be it at the low capability level of mutant from this work may be achieved in the medi- selection and breeding; at the medium level um term. of cell culture manipulation; or at the highest Some research teams in the more ad- level of difficulty, which is plant genetic vanced developed countries are undoubtedly engineering. At whatever level a laboratory technically capable of genetically engineering does its work, its impact is likely to be large pollutant-degrading microorganisms for since algal biotechnology is in its infancy, higher efficiency or a broader range of activi- leaving much room for adding knowledge to ty. However, questions about the possible algal biology and genetics and for developing risks that these organisms may pose to the applications for aquaculture and industry. environment would have to be resolved Since retraining a terrestrial plant tissue or before they are tested. Only then could coun- cell culturist for R&D focussed on marine tries formulate adequate regulations govern- plants could be done in three to six months, ing the field testing of the new organism. research projects in this area could soon be Because of the uncertainties posed by any under way. project that includes the release of genetically On the other hand, although algae are engineered organisms in the field and given plants, there are important differences be- the lack of regulations that govern such tween terrestrial plants and algae. The molec- Optlons In madrne blotechnology for developing countrdes 63 ular genetics of some plants, especially tobac- wonder, therefore, that research on marine co, has been intensively studied in cell and plants is as much as twenty years behind tissue culture for over ten years. To illus- terrestrial plants. Of course, some knowledge trate, one of the main difficulties in tissue from terrestrial plant basic research is trans- culture work was to regenerate a whole plant ferrable to marine plant research. Neverthe- from the callus. Intense study of the broad- less, much basic research needs to be done on leaved plants (dicotyledons), which include marine plants to clarify their growth charac- the tobacco plant, led to early success; scien- teristics; explain the molecular and genetic tists developed techniques that allow them to basis for regeneration; develop vectors to regenerate most dicotyledons. However, most introduce foreign genes effectively into ma- important crop plants are monocotyledons, rine plants; and so forth. After researchers and these plants were, and remain, very accomplish these goals, they will be able to difficult to regenerate. Nevertheless, respect- develop tissue and cell culture techniques that able progress has been achieved in the last reliably will make products of interest to two years including, for example, the regen- farmers and industrialists. Due to limited eration of rice. But the situation that today knowledge about marine plants, R&D that faces researchers investigating macroalgae is utilizes tissue and cell culture is likely to lead more complex and therefore more difficult. to applications, such as improved plant char- Unlike terrestrial plants, macroalgae do not acteristics, in the medium to long term. form seeds but instead rely on fragile spores The situation is, however, different for for propagation. Also, their life cycles are microalgae, such as Dunaliella, Porphyri- more intricate in that they may alternate diwn, and Spirulina. Because these organisms between sexual and asexual forms (Singleton have simpler structures than macroalgae, they and Kramer 1991). For these reasons there is are easier to investigate. Consequently, we much less fundamental knowledge about know more about their biochemistry, genetics tissue and cell culture of macroalgae than and physiology. In addition, while farming about many terrestrial plants. these organisms to produce beta carotene, Even more remarkable are the achieve- glycerol and health foods, culturists and ments of plant biotechnology. Vectors have industrialists have gained a vast amount of been developed for inserting foreign genes empirical knowledge about the large-scale into their genomes. Several plant species have propagation and processing of microalgae. been transformed by inserting functional Scientists in developing countries could adapt foreign genes into their genomes. For exam- or develop microalgae techniques for local ple, scientists have been able to genetically production of these substances in the short engineer a plant to resist pests by placing a term. In the medium term, research units gene coding for insect toxin in the plant's should be able to recover microalgae from genome. Conversely, the molecular genetics indigenous sites, screen them for promising of algae is new and no vectors have been substances, and do the applied research that developed for reliably transferring genes to would lead to the large-scale propagation of algae (Chapman and Gellenbeck 1989). promising new strains in cell culture. An immense amount of basic research has been done on terrestrial plants, which has laid Biosensors a basis for developments leading to applica- tions. To a great extent, this situation reflects As biosensor techniques are integral to the the amount of effort that has been devoted to performance of advanced research in biotech- terrestrial plant R&D, which is about 100 to nology, many research teams in developing 200 times that given to marine plants. It is no countries are able to construct monoclonal/ 64 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES polyclonal antibodies and manufacture DNA tries, if any, have the science-based industry probes. In the laboratory researchers employ necessary for designing, packaging and mar- them to detect and identify peptides, sequenc- keting detection kits based on monoclonal/ es of DNA, and other molecules of interest. polyclonal antibodies or DNA probes (see the When one moves from the laboratory next chapter). For these reasons, even though setting to the field, things become more some researchers are capable of developing difficult. Field detection activities are likely biosensors, we know of no facility in a devel- to be done by persons of varying ability and oping country that possesses the advanced training; highly variable physical forces act technology required to produce detector kits. on reagents and operators; and confounding The development of chemoreceptor and factors may cause false positives or, con- immunological biosensors is in its infancy. versely, conceal true positives. The ability to Some systems work well in the laboratory detect and track a sought-after chemical or performing specialized tasks. But extending organism in the field is largely dependent on their use past the laboratory stage requires operators having sensors/detectors that give further complicated, lengthy and expensive accurate, reproducible results, are simple to development. As a result, biosensor develop- use, have long shelf lives, and are minimally ment is a risky business venture; one that will effected by changes in light, temperature, not attract many investors. To sum up, it is humidity. The biosensor, whether a mono- unlikely that chemical or immunological clonal/polyclonal antibody, DNA probe or biosensor will available for general use in the another material, by itself is insufficient. short or medium term. Instead a detection kit is required, consisting of the biosensor, utensils to hold the sample, Biological oceanography reagents to allow the detector and sample to including public health react, reaction vessels or tubes, a read-out device, and positive and negative controls. One consequence of the public's concern All reagents and implements must be pack- about environmental degradation is that aged correctly and adequately protected from research in this field has increased tremen- rough handling and harsh meteorological dously. Microbial ecology is one of the conditions. research areas that have benefitted from this Thus, R&D to develop the detection kit trend, expanding at an exponential rate in has two major steps. The first, which is to industrialized countries. However, microbial construct or manufacture the biosensor (the ecology is a new discipline, one that is just monoclonal/polyclonal antibody or DNA beginning to generate information about probe), can be done in several developing interactions between humans, microbes and countries; research teams in other countries the environment. Given its basic research could gain the requisite capabilities in short orientation, it is not clear whether microbial order. Consequently, several nations are able ecology will experience the same rapid and to undertake the R&D that will lead to the expansive growth in developing countries as realization in the short term of biosensors for it has in the industrialized world. In some detecting pathogens or chemicals in marine cases, governments in developing countries environs. However, these biosensors would will indeed recognize the importance of this function efficaciously only when operated by scientific discipline and will promote its skilled researchers in the laboratory. growth in word and in deed. Even under such Leaving aside here the major problem of favorable conditions, it probably would take raising capital, very few developing coun- about two years before national researchers Options In nmarine biotechnology for developing countries 65 would receive the training to implement work freezing. This property, which is coded for programs in this area. The research projects by a single gene, may be successfully trans- undertaken by these newly trained researchers ferred with today's technology, although its would probably not generate applicable re- p-acticality still has to be proven. Other sults until the long term. However, in the conceptually attractive projects involving the meantime scientists from developing countries transfer of genes coding for useful properties trained in biological oceanography would be in marine plants to terrestrial plants can be well placed to help their governments make visualized. For example, marine plants have significant contributions to international evolved exquisite mechanisms to protect meetings, conferences and other fora where themselves from salt, so in theory it would global issues having high scientific content make sense to screen plants species possess- are considered and international relief mea- ing protective mechanisms against salt and sures are designed and put into practice. The assess whether they could be transferred to importance of this point, which is further terrestrial plants. While attractive in concept, elaborated in the next chapter, should not be for the present this type of project is today underestimated. beyond our scientific/technical capabilities In regions and countries where biological because it would involve the interactions oceanography and microbial ecology are not between many genes. Similar to the discus- bestowed high priority, important natural sion above on options in cell and tissue phenomena affecting human health and the culture, much basic research needs to be done environment will remain largely obscure for on marine plants before it will be possible to the foreseeable future. apply their important traits or characteristics on land. In particular, science needs to clarify Marine biotechnology applications the genetic control over metabolism in marine in terrestrial agriculture plants, including the production of secondary metabolites and tolerance to stresses, includ- Certain characteristics exhibited by marine ing salt stress. Thus, applications cannot be organisms may be utilized on land. We have expected in this area in the short or medium seen, for example, how the flounder anti- term. freeze protein seems to protect tomatoes from 5 Building capability in marine biotechnology A country's scientific establishment must termed basic research; it is usually character- have a capability in biotechnology generally ized as being performed without having before it can enter more specialized fields, specific applications in mind and most often such as marine biotechnology. While recog- is done at institutions of higher learning. nizing that a small number of developing Goal-oriented research is termed applied countries, including Argentina, Brazil, Chile, research; it is typically performed in national China, Cuba, India and Thailand, are per- laboratories, industry-based laboratories and, forming complex research in some areas of more rarely, at universities. Its purpose is to biotechnology, most have much more limited find out if something that works in the labo- capability. Therefore, a nation wishing to ratory will also work on a larger scale. If it build capability in biotechnology must: (1) does, the concept is further developed in a enhance present R&D capabilities or develop pilot plant, then in full-scale industrial pro- new capabilities in the biological sciences; (2) cesses. set up mechanisms for effectively transferring The general requirements for capability- research findings to the agricultural, industri- building in biotechnology research have been al or health sectors; (3) encourage indige- set forth in two reports (McConnell and nous, existing industry to develop the capaci- others 1986; NRC 1990); the more specific ty for applying research; and (4) promote requirements for marine biotechnology re- new biotechnology-based industry. Successful search are discussed in two UNIDO publica- interlinking of these four sets of activities can tions (Colwell 1986; Singleton and Kramer be achieved only if the home government 1991). An underlying assumption of capabili- gives high priority to capability building, then ty building in biotechnology is that the stan- takes the lead by formulating appropriate dards by which excellence in education and policies, committing the necessary resources scientific endeavors are measured and judged over the long term to implement them and, if do not change from one place to another. progress stalls, encouraging scientists and Circumstance fluctuate, priorities may differ, scientific administrators to continue their methods for reaching objectives vary, but the efforts by bestowing appropriate rewards. fundamental criteria of excellence in research and development do not change. Therefore, Building R&D capability the same factors apply in capability building, whether it occurs in an industrial or a devel- The process whereby a concept (or idea) oping country (McConnell and others 1986). is transformed into a finished product has This being so, capability building in R&D been called the concept development process anywhere depends, in the first instance, on (Zilinskas 1989). Capability building encom- well-educated, well-trained scientific person- passes all the stages of the concept develop- nel. ment process, beginning with basic research The schooling of scientists is long and and ending with the manufacture of products. demanding. Experience shows that it takes Research may be done to advance knowledge approximately twenty-five years (from prima- and to develop new products and processes. ry school through post-doctoral studies) to Research that emphasizes the first may be educate and train a biotechnology researcher, Buflding capability In marlno biotachnology 67 able to perform independent, advanced re- age them to stay and use their ability in their search. A sound basis has to be laid in sec- own countries. ondary school and undergraduate studies in The physical elements for good working mathematics, expository writing, reading, and conditions include: the natural sciences or engineering. In view of the interdisciplinary character of biotech- * Adequate facilities housing laboratories nology, students need a broad background in and associated structures, including aquar- the natural and physical sciences, as well as ia. mathematics. Thus study should include * Equipment of sufficient quality and quanti- general biology, cell biology, biochemistry, ty to do the research scientists have been microbiology and genetics; the future marine trained to perform. biotechnologist would find certain electives * The ready availability of common and rare useful, including aquaculture science, botany, chemicals needed to carry out experi- phycology and zoology. The chemistry cur- ments. riculum should include general chemistry, * A dependable infrastructure for sustained quantitative analysis, organic chemistry, and electric power, high quality water, and physical chemistry. Mathematics should gas. include statistics, calculus and differential * Well equipped and maintained equipment equations. Chemical engineering courses are repair and maintenance shops. essential for the engineering students; knowl- * Libraries containing a certain minimum edge in the computer sciences is essential for number of current books and journals. all students. In graduate training students plan * Computer-facilitated access to internation- and execute an independent research project al databases and computer mail (E-mail). to develop the ability to think through prob- lems and design experiments to solve them. In addition, there are less precise, but equally Frequent interactions with other students and important factors: critical readings of the relevant scientific literature are necessary to gain advanced * Salaries for scientists and technicians knowledge and to develop sound judgement. commensurate with their training and Scientists should do post-doctoral work in a abilities. laboratory different from her or his home * Possibilities for professional advancement institution to gain new skills and perspectives. based on ability and quality of work. Similarly, engineers should obtain practical * Availability of sufficient time and funds so experience at up-to-date pharmaceutical, that the scientist can communicate elec- chemical, or genetic engineering companies. tronically with colleagues in the interna- Once scientists have completed their tional scientific community and meet with training, the employment conditions under them while attending international meet- which they work are of paramount impor- ings and workshops. tance. The reward system for scientists is * Relief from unnecessary administrative or quite complex. Essentially, those who study bureaucratic procedures burdens. long and hard to become scientists rightfully * An equitable peer review system to review expect to use their talents fully, to advance a scientist's project proposals, ongoing intellectually, and to be paid a fair wage. In work, promotion and tenure. particular, scientists from developing coun- * Possibility for an individual scientist to be tries who receive advanced training must be part of a "critical mass" (see below). able to utilize these skills in order to encour- * Support by well-trained, highly motivated 68 MARINE BIOTECHNOLOGY AND DEVELONG COUNTRIES technical and secretarial support staff. or three different disciplines. A team * An equitable teaching load; that is, one undertaking research in medical biotech- that allows sufficient time for research. nology would consist of five to ten spe- * Individual incentives, such as small cialists in biochemistry, cell culture, grants. genetics, monoclonal/polyclonal antibody construction, microbiology, and molecular A significant research effort in biotechnol- biology. Depending on personal preferenc- ogy is best undertaken by an interdisciplinary es, each scientist should be supported by team, consisting of a sufficient number of three or four assistants, technicians and/or scientists and engineers to form a critical graduate students. mass: * Whether the team is expected to carry out The principle behind the idea of "critical basic research besides applied research. mass' is that five biotechnologists work- Scientists at university laboratories may, ing in the same place are more effective for instance, be expected to perform basic and productive than the same five biotech- research, or research aimed at solving nologists working in five different places. fundamental problems or adding to the This is related to the benefit derived from store of fundamental knowledge. Some- frequent interaction and help among bio- times basic research is needed to solve technologists, which is much easier and problems that arise when carrying out more effective when they are working in applied research or in development. Nev- the same place .... In order to use different ertheless, for whatever reason it is done, concepts or techniques to solve a certain it will take time and effort, which must be problem, it is essential that a team consist considered when determining the size and of biotechnologists from a range of disci- composition of the research team. plines. For example, a successful team of workers in biotechnology must have at its * Whether the team will have a training disposal several independent scientists in function. Obviously, if a scientist has each of the following basic disciplines: teaching and training duties, less time will biochemistry, microbiology, immunology, be available for research. The more inex- cell biology, and biochemical engineering. perienced the student, the greater the Depending on the type of project, a team effort the scientist needs to devote to may also need one or several experts in teaching. However, if the training period specialized fields such as genetics, virolo- is long, say two years, then the trainee gy, pharmacology, plant science, animal can be expected to contribute and be a net science, or food science (Wu 1986). asset after about one year. As a rule of thumb a scientist helped by four to five There are three main considerations when technicians could take on one or two long- attempting to calculate the optimal size of a term trainees without seriously handicap- research team: ping research. * The number of projects the team is ex- Equipment and material requirements for pected to take on. If one or two larger a well-furbished laboratory have been de- projects are being taken up, a critical scribed elsewhere (BOSTID 1982; Limonta mass for research in, for example, agri- 1989), so there is no need for a detailed cultural biotechnology can be achieved discussion of them here (a summary is pro- with five Ph.D.-level scientists from two vided in Appendix B). Bulding capaebty In madne biotechnology 69 Applying research present information about potentially market- able products to those who may wish to Somewhere along the concept development develop them. process, the concept is transferred from the From the industry side, the many compa- research laboratory to the development and nies that could conceivably benefit from testing facility. This step is vital, but due to introducing new or improved biotechnology systemic barriers that prevent technology techniques into their processes should make it transfer from university to industry, it takes a high priority to set up advanced develop- place but rarely in developing countries mental laboratories. These counterparts to the (Zilinskas 1989). Each side, university and university technology transfer unit would industry, must initiative certain actions that have four responsibilities: to seek out re- overcome barriers; somewhat like two crews search being performed in public laboratories constructing a tunnel, working from opposite that could be applied by the firm who they sides of a mountain, but each doing what it work for; to negotiate with the public labora- must to reach the other. tory for rights to use these results; to perform The major indigenous technology produc- the advanced development that would enable ers in the developing countries are universi- the firm to utilize research results effectively; ties and public scientific or technical insti- and to contract with the public laboratory for tutes. It is imperative that they take the initia- further research required to solve problems tive to make certain that the research results or improve processes (Zilinskas 1992). they engender, and the inventions their scien- tists conceive, reach those who can apply Industry and biotechnology them. The best way to do so is for them to establish technology transfer units (Zilinskas It must be made clear that research ac- 1992). Often, this will mean that universities complishments will remain tantalizing prom- will have to break out of the mold set per- ises unless someone, or some entity, accesses haps hundreds of years ago, that of the aca- its results and develops them. Results from demic ivory tower. Universities are changing biotechnology research can theoretically everywhere, even the ancient universities of benefit technology end users in established, Europe that once epitomized the academic traditional firms and new biotechnology-based ivory tower. One reason for this change is industries. The first type of firms exist that the scientific research done at these throughout the world, manufacturing food universities can no longer be designated as and chemicals or extracting natural resource. basic (as contrasted to applied research done However, by far most firms in developing usually by industry). In biotechnology espe- countries have capabilities only to manufac- cially, findings from so-called basic research ture, package and market goods. They do not can have almost immediate applied implica- possess have applied research or advanced tions. For example, when a researcher clari- development units, which limits their ability fies the molecular control in a cell that pro- to absorb or adopt results from indigenous or duces a protein, he or she is at the same time foreign scientific research. They must there- mapping out a production process that is of fore act to set up advanced development lab- interest to industry. Unless the researcher, oratories, as discussed in the previous para- and the university employing that researcher, graph. is willing to forego a possibly significant The second type of enterprise, the biotech- financial reward, the university must track nology-based company, is common in the the research being done at its laboratories, industrial countries, but only a few of them assess its applied impacts and aggressively have been established in developing coun- 70 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES tries. For example, in Peru there has been Beginning with the first step in the con- only one biotechnology company, Bio- cept development process, research at univer- ingenierfa Aplicada. This was also the first sities and institutes in developing countries is company in Peru to set up a direct link be- supported almost entirely by governments and tween industry and university; and for a time researchers are mostly government employ- Bioingenierfa Aplicada was able to exploit for ees. As is well known, public universities in profit the indigenous natural resources of that most developing countries are underfunded country on a sustainable and environmentally and researchers are underpaid (when com- sound basis, but the company went out of pared to persons in the private sector who business in early 1992. have approximately the same education and It is important to encourage the estab- responsibilities). In a time of severe budget- lishment of biotechnology companies in the ary constraints it is difficult to correct these developing countries because they will proba- shortcomings; nevertheless, it must be done bly be the main users of research results from before a country can build the indigenous universities and the most impor- scientific/technical base upon which a sci- tant vehicle for the development and commer- ence-based industry, such as biotechnology cialization of biotechnology products in their industry, can grow and flourish. Accordingly, countries. However, as is the case of technol- a significant strengthening of research can ogy transfer units in industry, governments only come about if governments take the hard have the major role in creating the economic decisions to raise new funds or divert scarce climate conducive to the entrepreneurship of funds from other programs in order to streng- biotechnology-based industry because only then scientific institutions and finance neces- they can adopt measures that encourage sary research. people to make the risky investments required Supporting research and researchers is, to establish that industry. however, only the first of several activities a government must support before a concept is The role of government in building transformed into a product. After the scien- capability in biotechnology tist-inventor has conceived a concept, the concept has been researched, and it has been An analysis of the factors affecting the verified as commercially promising in the development of biotechnology showed that laboratory, a point is reached where funds for the three most important are government the advanced development or the concept is funding of basic and applied research, scien- required. This point is crucial because devel- tific personnel availability and the level of opment will cost on the order of seven to ten their training, and the availability of financ- times as much as research. However, devel- ing and tax incentives for biotechnology opmental funds are exceedingly difficult to business. Next in importance was the exis- raise since investors will perceive a venture at tence and adequacy of health, safety and this early stage of development as being a environmental regulations, the operation of an risky investment. A government is probably equitable intellectual property law, and effec- the only source in most developing countries tiveuniversity-industry relations (OTA 1984). for this kind of funding; if it is not available, The most cursory analysis of these factors the budding venture dies. Accordingly, gov- demonstrates the vital role of government, to ernments must make funds available to indus- make certain that economic, legal and politi- try, either as grants or low-cost, long-term cal conditions favoring capability building are loans, so it can set up advanced development present in the country where it is being done. laboratories and pilot plant operations. Bulding capability In marine biotechnology 71 Tax laws may be designed to encourage formulated by OECD (for a general discus- investments in R&D, which is very important sion of the subject of biosafety and develop- to science-based businesses. Governments ing countries, see Trigo and Jaffe 1990). may do this through three methods. First, However, the biosafety issue is problematic laws may be designed to promote investments when we move past research. Most develop- in R&D by allowing the company that invests ing countries have not promulgated regula- its profits in R&D to credit a proportion of tions that specifically address biotechnology this amount against owed taxes. Second, research, development, testing or manufac- investors who invest in risky R&D should be ture. In some cases, laws or regulations that able to credit losses occurred in doing so deal with the environment, occupational against owed taxes. Third, tax laws should be health or industrial activity may have implica- designed so they do not unduly tax profits tions for bioscientific or bio-industrial activi- from profitable investments, thus appearing to ties, but national agencies will not or do not penalize success. enforce them because they lack the expertise There has been much debate on the pro- or resources to do so. In view of this uncer- priety of patenting life forms and the fairness tain situation in most developing countries, of allowing patents on procedures having companies will be unable to adequately plan wide applicability, such as the PCR (Korn- for the development, testing and manufacture berg 1991). This is not the place to debate of biotechnology products. If they are unable these issues, it is however appropriate to note to perform these vital activities, they will in that investors are not likely to invest capital effect be precluded from entering into bio- to develop inventions unless they are assured technology R&D in most developing coun- of exclusive, long-term rights to those inven- tries. Accordingly, governments must adopt tions. Further, companies, whether multina- equitable and adequate biosafety regulations tional or national, are unlikely to develop that govern research, development, testing new or unique products in countries where and manufacture. This will have the effect of they cannot protect these products adequately creating the stable regulatory climate neces- through licensing or patenting. Accordingly, sary for industry to do long-term strategic governments cannot ignore the intellectual planning. property issues as they concern biotechnology inventions. If governments wish to set up Discussion biotechnology-based industries, they will find it well worth their while to consult with the While keeping in mind the concept of the World Intellectual Property Organization in development process, and its starting point in Geneva on this matter, and to analyze the research, this report emphasizes the applica- Budapest Treaty on the International Recogn.- tions of marine biotechnology. For one, the tion of the Deposit of Microorganisms for the term "biotechnology" implies application. Purposes of Patent Procedure in terms of its Further, application is the goal of most assis- own national interests. tance provided by the World Bank to devel- Usually research is not effected by lack of oping countries. However, this should in no regulations since universities are often auton- way be interpreted to mean that basic re- omous in developing countries, enabling them search is unimportant to, or an unnecessary to adopt their own rules governing R&D. In luxury for, developing countries. There are addition, they can easily adopt rules promul- three major reasons why this is so. First, the gated elsewhere such as, for example, the basic research typically performed at teaching NIH guidelines or the generic regulations institutions provide training in fundamental 72 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES techniques to the scientific/technical person- ment promoting contacts between the marine nel who will, in turn, teach and train students natural science researchers and the field at secondary schools and universities, staff workers in aquaculture, fisheries and other biotechnology industry and firms that utilizes industries. biotechnology techniques, and give expert Besides the factors that directly affect the advice to governments. Second, in the course development of options for developing coun- of developing a concept, especially at the tries in marine biotechnology, an indirect pilot plant and manufacturing stages, prob- factor bears mentioning. A useful side-effect lems often arise whose solution requires basic that accrues to developing countries that have research. Third, basic research is a type of attained significant capabilities in marine "safety valve" to imaginative scientists, biotechnology is that their level of expertise providing them with the mode for expressing in marine and oceanographic biology will their creativity that at times engenders unex- simultaneously increase markedly. In particu- pected results important to the applied sec- lar, as capability building encompasses train- tors. Clearly, any country that wishes to have ing, more scientists in developing countries an in-depth, encompassing bioscience and become trained in biological oceanography biotechnology infrastructure requires a strong and marine biodiversity; individuals who basic research component in the biological thereafter will be well-placed to perform the and biochemical sciences (McConnell and investigations and technological assessments others 1986). that will form the basis for the formulation of Nevertheless, as shown by the historic national policies and regulations to protect example of development in Japan and, more and manage indigenous marine resources. recently, by Hong Kong, Singapore and the Further, these experts would be well Republic of Korea, a country that emphasizes equipped to serve as consultants to national applied R&D and, concurrently, constructive- delegations at international meetings and ly imports and adapts foreign technology is conferences whose aim will be to deal with likely to benefit through accelerated economic the consequences of human activities that development. Perhaps similar measures can interfere with, modify or disturb natural be taken by countries rich in marine resourc- processes. In particular, since the oceans are es, who have the option of capitalizing on the final depository for many pollutants and these resources by investing in marine bio- serve as the world's major carbon sink technology. The main ingredients to enter this (Initiative 1990), input from scientists knowl- field are often present or are readily attain- edgeable about marine natural sciences will able: biotechnology is relatively accessible to be required when politicians formulate inter- the scientific community since its techniques national measures to mitigate the negative have been published or may be accessed in effects stemming from the 'greenhouse' data banks; the universities of most countries effect, the destruction of ozone in the strato- have ongoing teaching and research programs sphere, the increasing levels of toxic agro- in biology, biochemistry, marine biology and chemicals in ground water and oceans, and the ocean sciences, which provides a scientif- other problems that know no national bound- ic basis for biotechnology; and a technical aries. As a result of them having the requisite basis exists as well in many countries in the expertise in the marine biosciences, develop- form of established aquaculture, fisheries and ing countries will be able to contribute sub- natural products development. If these ingre- stantially to the crafting of international dients are present, it is possible that a country measures that seek to alleviate present global can benefit in the short term by its govern- problems and prevent future ones. 6 The experience of major intemational agencies in marine biotechnology and related areas Major international organizations and mission, the FAO has since its inception some nongovernmental organizations (NGOs) provided major funding to developing coun- were contacted and requested to provide tries for investments in fisheries equipment information about their activities in marine and for strengthening their management biotechnology and marine biotechnology- capabilities. Significant FAO funding has also related areas. Some agencies provided much been directed at expanding aquaculture in information about their work, others did not developing countries. In all, the FAO spon- respond or gave only sketchy information. So sored 114 aquaculture projects during the amount of space allocated to agencies in 1972-1984 at a cost of approximately $51 the following sections do not necessarily million (FAO 1991). All were aimed at the relate to the size or extent of their programs production of food; few included research. Of in marine biotechnology-related areas. the thirty projects that did include a research We stress that most of the information we component, nine used laboratory facilities, present below was provided by the agencies but only one consisted solely of research. In themselves; in some cases we were able to general, research is done to improve aquacul- supplement it through interviews with agency ture systems, to identify new professionals administering pertinentprojects. species/aquaculture system combinations, and From the responses we received it is clear to identify indigenous species that may be that with the exception of UNIDO, no organi- suitable for aquaculture. zation has experience in marine biotechnolo- FAO is aware that many fisheries have gy per se, but several are supporting or are reached their maximum sustainable yield, and involved in marine biotechnology-related that some are beyond that stage. Beginning in areas. In this chapter we describe these activ- 1989, the FAO's Fisheries Department has ities and attempt as possible to orient them therefore greatly expanded its efforts in within the agencies' overall work program. aquaculture. In particular, it is taking a criti- The agencies are dealt with in alphabetical cal look at the future of aquaculture develop- order. Each agency is covered in a section ment, including culture methods and siting that has two parts. The first describes the problems (FAO 1989). In this vein, it is agency's activities in marine biotechnology- sponsoring the Seafarming Development and related areas. The second part contains our Demonstration Project of NACA in Thailand, thoughts on possible ways that the agency can which aims to overcome problems that beset incorporate marine biotechnology in its work aquaculture, including seed supply, culture program. We end this chapter with a brief techniques, post-harvest processing and the discussion on the reasons why professionals training of aquaculturists (Lovatelli 1990). of agencies are mostly uninformed about Among its regional initiatives, it has set up marine biotechnology. the UNDP/FAQ Regional Seafarming Devel- opment Project, which sponsored the first Food and Agricultural Organization Asia-Pacific Regional Workshop on the Culture and Utilization of Seaweed, held in The major goal of the FAO is to increase Cebu, Philippines during August 1990. the world's food supply. In line with this The FAO indicated that it is not sponsor- 74 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES ing any marine biotechnology projects. It has, logy's long history in the South East Asian however, been sponsoring some research in countries, projects amalgamating aquaculture related areas, such as marine biology and and biotechnology seems feasible. If such aquaculture, and is currently developing its projects were developed, the ADB would understanding of population genetics and probably be interested in providing loans to biodiversity of wild and cultured species. If fund them. these approaches were to be further devel- The European Investment Bank (EIB), the oped, it could open the way for a FAO pro- European Bank for Reconstruction and Devel- gram that would include marine biotechnolo- opment (EBRD), the Andean Development gy. In addition, as is made clear above, Corporation (Corporaci6n Andina de marine biotechnology may be used to signifi- Fomento), and the African Development cantly increase yields from aquaculture Bank (AFDB) are, or have been, involved in through the genetic improvement of culture aquaculture projects. In addition, they are organisms and by improving health among increasing their support for biotechnology cultured species. These possibilities relate projects. These regional banks would proba- directly to FAO's mission of improving food bly assess proposals for marine biotechnology supplies and might therefore be taken up by projects like any other project; that is, in the agency. But as the situation now stands, terms of their likely return on investment. it is unclear whether FAO will undertake significant initiatives in marine biotechnology Intergovernmental Oceanographic in the near term. (FAO's involvement in bio- Commission safety is discussed below in the UNIDO section.) The IOC, established in 1960, is a func- tionally autonomous (semi-independent) Inter-American Development Bank agency, which means it has its own member- and other Development Banks ship, statutes and governing body, even though its secretariat is sited at the headquar- The IDB has done extensive lending in the ters of the U.N. Education, Scientific and field of fisheries. Many of these loans are Cultural Organization (UNESCO) in Paris. directed to private fishing fleets and fish The IOC promotes marine scientific investi- processing plants. While there are a few gations and systematic ocean observations aquaculture and research oriented activities through concerted actions in partnership with funded by IDB, the overall picture is one of its member states and other international limited funding in sectors related to marine organizations. The specific activities the IOC biotechnology. The IDB would probably be is involved with include supporting the global interested in providing loans for activities that sea-level observing system (GLOSS); the would improve current aquaculture practices joint IOC-World Meteorological Organization or set up marine natural product development program called Integrated Global Ocean (Peacock 1991). System (IGOSS); ocean science research on The Asian Development Bank (ADB) has living resources in cooperation with FAO; been investing in aquaculture and fisheries in research on nonliving resources with various countries like Bangladesh, Burma, Indonesia, U.N. organizations; marine pollution investi- Pakistan, Sri Lanka and Thailand. The GIFT gations and monitoring in cooperation with project at ICLARM is an example of such UNEP and the International Atomic Energy projects (see discussion in the section on Agency through the Global Investigation of ICLARM, page 82). In view of biotechno- Pollution in the Marine Environment pro- Ma/or international agencies and blotechnology 75 gram; and the global mussel watch (see page consisting of thirteen eminent scientists, 4). An important new initiative is the devel- assists and advises the Committee and the opment of the Global Ocean Observing Sys- ICGEB Director, Dr. A. Falaschi. Fifteen tem (GOOS), which aims to collect data on research centers have been designated as climatic variability and change to be used to ICGEB affiliated centers: in Algeria, Argenti- construct prediction models. Training, educa- na, Brazil, Bulgaria, Chile, China, Cuba, tion, mutual assistance and partnerships are Egypt, Greece, Hungary, Mexico, Nigeria, important elements in all IOC activities. The Tunisia, Venezuela and Yugoslavia. Applica- long-term goal is to develop IOC programs tions from Iran and Turkey are pending. that will permit management of oceans simi- Since more than twenty-four countries have larly to how we today manage agricultural ratified the ICGEB's statutes, the Centre is systems. poised to becomes a free-standing intergov- IOC activities to date do not bear on ernmental organization. This development is marine biotechnology, but the development of likely to occur in early 1993. Until that time, biological detection systems might use ad- ICGEB is run by UNIDO as a project accord- vanced marine biotechnological methods. For ing to a rolling five-year program. At pres- example, IOC's marine pollution research ent, ICGEB is funded at $72 million for the program includes studies on the biological period July 1992 through June 1997 (ICGEB effects of pollution and the development of 1990). observation and statistical methods that may Of indirect interest to this report is be used to monitor changes; this program ICGEB's involvement in biosafety. Specifi- would benefit from the employment of bio- cally, a joint UNEP/ICGEB program on sensors. There is also a strong desire at IOC biosafety commenced in 1991. Its two initial to coordinate several oceanographic activities activities consisted of courses offered in to enhance our understanding of living and Trieste to researchers from developing coun- nonliving marine systems. Since marine tries: a three-day course was held July 1991 biotechnology opens added opportunities to and attended by thirty scientists called "Ge- further understand marine environments, netically Modified Organisms: Safety in the there exist possibilities for cooperating on a Laboratory and the Environment" and a variety of marine research related activities subsequent three-day course with fifty partici- (Kullenberg 1991). pants called "Genetically Modified Organisms for the 1990's" followed. International Centre for Genetic While the ICGEB is not presently under- Engineering and Biotechnology taking projects in marine biotechnology, it has offered courses to scientists of member Beginning in 1981, UNIDO was instru- countries related to the field, including one mental in establishing the ICGEB, which is presented December 16-20, 1991 called now operational in two components, located "Marine Microbiology and Biochemistry". in New Delhi, India and Trieste, Italy, and Given its five year program cycle, in the including as an integral part a network of short to medium term the ICGEB is not likely affiliated centers in countries around the to become deeply involved in marine biotech- world (Taylhardat 1989a and 1989b). The nology. It probably will continue offering ICGEB's governing body is its Preparatory courses on techniques and processes that Committee, made up of representatives from relate to marine biotechnology, thereby the forty-five countries that have signed the helping countries whose scientists attend to Statutes. A Panel of Scientific Advisers, build capabilities relevant to the field. In 76 MARINEBIOTECHNOLOGYANDDEVELOPING COUNTRIES addition, as scientists in developing countries United Nations Conference on learn the techniques of risk assessment and the Environment and Development risk management at the ICGEB, they will be able to assist their governments in formulat- UNCED, also known as "Earth Summit", ing a regulatory regime that allows a biotech- was held during June 3-13, 1992 in Rio de nology-based industry to operate. As we Janeiro, Brazil. UNCED's major objectives noted above, adequate regulations is very were to suggest steps that may be taken by important for the concept development pro- governments and international organizations cess to function in a country. to alleviate the damage done to the environ- ment by human activities and to prevent International Maritime Organization future damage, while allowing economic development to proceed relatively unhin- The IMO, headquartered in London, dered. UNCED's Agenda 21, which is an focusses on shipping-related activities. We action agenda for the 21st century addressed have no indication that IMO sponsors marine to govermments, IGOs and NGOs, includes biotechnology or marine biotechnology-relat- the item "Environmentally Sound Manage- ed projects. ment of Biotechnology". In the course of our research we have UNCED's Preparatory Committee found out that various agencies are preparing (PrepCom) was established in early 1990; it projects under the Global Environmental had three working groups. Working Group 1, Facility (GEF) that bear on ports' reception chaired by Mr. Bo Kjellen from Sweden, facilities; some of these include bioreme- addressed a variety of important issues, in- diation activities in developing countries. cluding the atmosphere, forests, land resourc- Apparently, IMO will be one of the agencies es, desertification, biodiversity and biotech- consulted for such work. Further, IMO has a nology; Working Group 2, chaired by Dr. good knowledge of the needs of waste dispos- Bukar Shaib from Nigeria, addressed oceans, al facilities and its relation to the shipping freshwater resources and the management and industry and is experienced with the problems movement of all types of wastes; and Work- created by oil spills. It would therefore seem ing Group 3, chaired by Mr. Bedrich Moldan that it would be in the agency's interest to from Czechoslovakia, addressed crosscutting consider the value of supporting three sub- issues, including legal and institutional issues, areas of marine biotechnology: bioreme- human settlements, technology transfer and diation of ship wastes, bioremediation of oil Agenda 21. spills and biofouling. Once involved with Chapter 16 of Agenda 21 identifies five enhancing bioremediation, IMO could support objectives for activities to promote biotech- R&D that attempts to adapt bioremediation nology, especially in developing countries: techniques for the treatment of various haz- increase the availability of food, feed and ardous wastes and raw sewage that would renewable raw materials; improve human otherwise be disposed of in the oceans. In health; enhance protection of the environ- addition, it might consider the question of ment; enhance safety and develop interna- replacing paints used to prevent the bio- tional mechanisms for cooperation; and fouling of ships' hulls, but which generally establish enabling mechanisms for the devel- are very toxic to marine life. If biological opment and environmentally sound applica- films that prevent the propagation of a variety tion of biotechnology (UNCED 1992). In of sea organisms would be developed, pollu- addition, there should be an explicit link tion of harbors and intensely trafficked waters between biotechnology and biodiversity, could be reduced. covering the forest, soil, freshwater and Major international agencies and blotechnology 77 marine sectors (UNCED 1991). cate an interest in marine biotechnology and Marine biotechnology is not mentioned in would promote projects in this field that any of the UNCED documents. Perhaps the would enhance economic development in publication of this report will stimulate a recipient countries. For example, UNDP is movement to include this field while carrying investigating irrigation schemes that utilize out the other Agenda 21 biotechnology objec- nutrient rich sea water to grow salt water tives. In particular, marine biotechnology resistant plants. This type of project could could help connect biodiversity and biotech- benefit from the application of marine bio- nology. In the marine area, one of the best technology techniques. ways to do so is through biological oceanog- raphy (see pages 64 and following); that is, United Nations Educational, Scientific to use advanced biotechnology techniques to and Cultural Organization track the development and movements of marine organisms, especially microorgan- UNESCO set up a program related to isms; to clarify dispersion mechanisms for environmental and applied microbiology organisms and genes; and to trace the evolu- already in 1946; ten years later it established tionary development of marine species. In the Panel on Microbiology, which was addition, bioremediation techniques hold charged with setting up an international promise for cleaning up much of the degrad- network for the exchange and preservation of ed environment common to coastal develop- industrial microorganisms and to train micro- ing countries and for preventing future pollu- biologists. In 1970 UNESCO acted to estab- tion through appropriate treatment of wastes lish a network of Microbial Resource Centers from land and water-based sources. (MIRCEN), which was indeed established in 1975 in cooperation with UNEP and the United Nations Development Programme International Cell Research Organization. MIRCEN has since then expanded, it now UNDP, the major provider of technical includes twenty-three centers in nineteen assistance among U.N. agencies, has a long countries. In addition to its original aims of history of supporting aquaculture projects in maintaining cell culture collections, MIRCEN developing countries, including China, Cuba, now promotes international cooperation in India, Indonesia, Ivory Coast, Lao P.D.R., many aspects of applied microbiology and Madagascar, Nepal and Viet Nam. Some of biotechnology, provides specialized training these projects involving marine fish and to developing country researchers on ad- shrimp species include R&D whose objective vanced techniques of biotechnology, and in was to genetically improve stocks, for exam- general seeks to improve the quality of life in ple, the GIFT project at ICLARM (see page the developing countries through the applica- 82). Similarly, a project (begun in 1990 and tions of biotechnology. executed by FAO) to develop seaweed pro- In addition to supporting MIRCEN, duction methods in the Philippines has a UNESCO fosters inter-agency cooperation research component for improving algal through the organizing of periodic interna- strains. tional conferences, called Global Impacts of UNDP supports several projects in agri- Applied Microbiology (GIAM), that are held culture, health and industry that include approximately every four years; the last one biotechnology components. Conversely, the was in 1991 in Malta. GIAMs bring together agency has so far not been involved in ma- scientists, as well as decision makers, from rine biotechnology. Nevertheless, the UNDP around the world to consider how applied professionals that we have interviewed indi- microbiology/biotechnology may be directed 78 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES for the benefit of developing countries and to particularly its ten regional seas programs improve the environment. that cover Mediterranean, Kuwait region, Starting in 1990, UNESCO set up the Red Sea, Caribbean, the Atlantic coast of Biotechnology Action Council, which admin- West and Central Africa, the East African isters a research and training program in seaboard, the Pacific coast of South America, biotechnology related to terrestrial and aquat- the islands of the South Pacific, the East ic plants. During 1992-93 the Council ex- Asian region, and the South Asian Seas. pects to award nineteen short-term fellow- These programs include activities such as the ships, and in 1993 hopes to fund five profes- monitoring and control of marine pollution, sorships. as well as the coordination of pollution pre- In the marine field, UNESCO cooperates vention strategies. in the work of the IOC to promote marine As is the case with other agencies, UNEP scientific investigations (see above), including is not presently supporting marine biotechnol- the Coastal and Marine Program (COMAR); ogy projects. The question then is what help has established Man and the Biosphere marine biotechnology may offer the agency to Programme (MAB) to develop a basis for the fulfill its mission. One possibility of potential rational use and conservation of terrestrial importance are biosensors. The joint and marine resources; and is active in the IOC/UNEP Group of Experts on the Scientif- International Hydrological Programme, which ic Aspects of Marine Pollution (GESAMP) is developing a scientific basis for the man- has been exceedingly active in determining agement of water resources, including biore- some of the problems of marine pollution; its mediation of polluted waters. work could be enhanced through advanced While UNESCO has not been involved in biotechnology detection techniques. Another marine biotechnology, without doubt, possibility is for UNEP to investigate and UNESCO could make an important contribu- clarify the possibility of using bioremediation tion to capability building in marine biotech- to treat pollution in developing countries and nology R&D since it is the lead agency in the to adapt bioremediation techniques for the UN system for helping countries improve treatment of waste water from land based their educational institutions and basic re- sources. (UNEP's involvement with biosafety search institutes, and in view of its wide- is discussed below in the UNIDO section.) ranging programs related to biotechnology generally. In addition, UNESCO is well United Nations Industrial placed to identify counterpart institutions that Development Organization could cooperate in marine biotechnology research having international significance. It UNIDO's interest in marine biotechnology also could inform policy makers in coastal is of rather long duration, beginning in the and island developing countries about the early l980s. The agency approaches this field possibilities that marine biotechnology hold from two directions-from its multifaceted for them through UNESCO publications and biotechnology program and from its interest UNESCO-supported conferences. in setting up a marine sciences center. UNIDO supports numerous projects in biotechnology that focus on applied R&D and United Nations Environment Programme on pharmaceutical and industrial applications. Developments throughout the world in bio- Some of UNEP's most significant activi- technology, including its marine aspects, are ties are focussed on the marine environment, reported in the quarterly publication Genetic Major international ogencles and biotechnology 79 Engineering and Biotechnology Monitor, broad agreement among countries where which is sent free on request to scientists in advanced biotechnology R&D is done on the developing countries. However, UNIDO's level of control appropriate for research and interest in marine biotechnology became large-scale contained use of genetically engi- manifest when Dr. Rita R. Colwell was neered organisms. Similarly, products pro- commissioned to write the report Marine duced by conventionally developed or geneti- Biotechnology and the Developing Countries cally engineered microorganisms for con- (1986), which discusses the rich promises the tained use are accommodated under existing field holds for developing countries and national regulations governing drugs, food or outlines an approach whereby they can build environment. However, no such international the requisite capabilities to fulfill these prom- unanimity exists in regard to the testing in the ises. Dr. Colwell's ideas were expanded on field of genetically engineered animals, plants in 1989, when the report "Biotechnology of or microorganisms. Because of this predica- Marine Algae: Opportunities for Developing ment, two detrimental consequences have Countries' was published (Singleton and occurred. First, in one case when testing was Kramer 1991). A third report is now being done by a U.S. commercial firm in Argenti- written, on the specific uses that marine na, a dispute arose about the adequacy of biotechnology has for the Mediterranean safety precautions of the test. The motives of countries. the firm conducting the test were questioned, UNIDO is also setting up a program on the implication being that testing was being bioremediation and oil recovery. Primarily done where local regulations were weak or intended for countries belonging to the Orga- nonexistent. Without adequate biosafety nization of Petroleum Exporting Countries regulations, some fear that unscrupulous (OPEC), it will offer short and long term companies may seek to field test organisms training to scientists for improving strains of and products where there are few legal hin- oil degrading microorganism and using them drances. Second, and more commonly, be- for oil spill cleanup or enhanced oil recovery. cause regulatory agencies have been unclear Of relevance to marine biosafety is on how to proceed, they have held up the UNIDO's initiative to establish an interagen- field testing of potentially valuable products. cy working group on biosafety. In 1985 In recognition of the uncertainties sur- UNIDO asked UNEP to form a joint working rounding field testing, the Working Group group to consider safety aspects of biotech- has recommended that the agencies represent- nology research, bioindustry and field testing. ed on it should undertake three activities: The working group was set up in 1986 and has since then met five times. During the last * Formulate an International Code of Con- two years WHO and FAO joined it. The duct for the Release of Organisms Into the principle driving the UNIDO/UNEP/- Environment. UNIDO took the lead in WHO/FAO Working Group on Biosafety, as drafting the Code. A working group of it is now called, is that the guidelines it thirty-five experts met with the Working develops should strike a balance between the Group in June 1991 to formulate a draft need to protect workers, public and environ- code; the draft code was further improved ment and the exigency of allowing biotech- on, then adopted in July 1991 (UNIDO nology to develop effectively. 1991). As the Working Group's work has pro- gressed, and as it gains experience, its strate- * Establish the International Information gy has evolved. As mentioned above, there is Resource for the Release of Organisms 80 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES Into the Environment. UNEP is the lead treatment of a specific topic in marine indus- agency for establishing the Resource, trial technology; the second contains capsule which will collect and store all available reviews of topical developments in the field information about deliberate release activi- (including marine biotechnology) and a listing ties throughout the world. Its steering of technical conferences and meetings. Free committee was set up in March 1991; its subscriptions to the monitor may be requested first responsibility is to take an inventory from UNIDO's Industrial Technology Devel- of existing information sources on release opment Division. activities. While UNIDO has not defined a work program for itself in marine biotechnology, it Draft and publish a biosafety manual. A provides technical assistance for marine manual for principally the developing biotechnology-related projects and it has countries will be prepared jointly by made certain that biotechnology components UNIDO, UNEP, FAO and WHO for are included in the proposed marine sciences publication in 1992. It will contain a centers. In the first instance, R&D in the compilation of national and international components might focus its R&D efforts on biosafety rules and guidelines, which may bioremediation, especially since the centers be drawn upon when international will be sited in regions heavily influenced by attempts are made to harmonize biosafety pollutants from ocean and land-based sources. regulations. It will also seek to clarify the UNIDO might also expand its natural re- safe conduct of biotechnology research sources program, to include projects aimed at and its applications. the collecting and screening of marine organ- isms for bioactive substances and chemicals Referring to the second direction, in the useful to industry and in agriculture. marine sciences, unlike other agencies dis- cussed above, UNIDO focusses on industrial World Bank applications in the marine sector rather than the marine environment per se. UNIDO The point can be made immediately; there senior staff has since 1984 been laying a is no World Bank experience in marine basis for establishing two marine sciences biotechnology. In effect, this report introduc- centers of excellence for developing coun- es the World Bank to marine biotechnology. tries, one in the Caribbean and the other in Since this is the case, we have sought out the Mediterranean. The first, tentatively projects supported by the World Bank in named the International Ocean Institute may marine biotechnology-related areas, including be set up as early as this year. As now fisheries, aquaculture and general biotechnol- planned, a component of the institute will be ogy. Options for future World Bank activities devoted to investigating the industrial poten- in or related to marine biotechnology are set tial of marine biotechnology for developing forth and discussed in the next chapter. countries. As of this writing, support from The World Bank sponsored the project major donor countries is being sought. "Study of International Fisheries Research" In 1991 UNIDO began to publish quarter- during 1989-90, which was published in ly the Marine Industrial Technology Monitor, 1991 (World Bank 1991b). In view of the the fourth in a series of such publications (the importance of fisheries to many developing others deal with advanced materials, biotech- countries, this study was timely. To illustrate, nology and microelectronics). Each issue has out of the leading ten fishing nations in the two parts. The first consists of an in-depth world, six are developing countries (Chile, Mlor internatlonai agences and biotechnology 81 China, India, Indonesia, Peru and Thailand) support for the stocking of reservoirs in and one, the Republic of Korea, is a newly Brazil, and in 1991 a Malawi fisheries loan industrial country (1989 FAO statistics 1992). included components for aquaculture. One important objective of the study was to In terrestrial biotechnology, the major determine high priority research needs related support provided by the World Bank is relat- to fisheries and aquaculture, assess the capac- ed to agriculture R&D performed at the ity of certain developing countries to perform sixteen International Agricultural Research fisheries and aquaculture research, and to Centers (IARCs) of the Consultative Group formulate strategies for improving donor on International Agricultural Research support for research. In the course of the (CGIAR). Programs carried out by the Study seven technical papers were prepared CGIAR centers include research to increase dealing with fisheries and aquaculture R&D productivity of agriculture and animal hus- capabilities of many African, Asian and Latin bandry, management of natural resources and American countries, the development of germplasm preservation. The World Bank tropical aquaculture, international cooperation provided about 15 percent ($32 million) of in fisheries research, and research needs of CGIAR's 1990 budget of $235 million. small-scale fisheries. One finding of the study CGIAR's research purview includes com- was that there is among surveyed countries a modities that provide 75 percent of the food need for research aimed at the genetic im- energy, including protein, for the world's provement in cultured species to improve population (World Bank 1992). So far, health, growth and feed requirements. As a CGIAR research centers have been almost result of the study, the World Bank staff has entirely focussed on terrestrial resources. shown an increased interest in fisheries re- Further, by far most research done at the search; an activity that could easily include a CGIAR centers is applied, with the aim of marine biotechnology component. producing useful results in as short time as In general, World Bank activities in the possible. Traditional agricultural techniques aquaculture sector has expanded greatly in the are at the current stage of development the past decades. These activities might occur safest and most efficient systems for increas- either as a sub-component of larger fisheries ing the productivity of crops (as evidenced by or agricultural projects, or as aquaculture the 'Green Revolution") and the quality and projects with lines of credit. Alternatively, quantity of animal production. As a recent projects may be funded through multilateral World Bank supported project has found, the investment banks and donor organizations, biotechnology techniques more advanced than for example, in 1985 $14.1 million was tissue culture have for this reason not been provided in this way. In this case, there is a extensively adopted in CGIAR laboratories divisions of competence between the World (World Bank 199 la). The more sophisticated Bank and some of the regional banks, that is, genetic engineering methods employed by ADB and IDB, so that the latter institutions modern biotechnology are gradually being finance the bulk of the aquaculture projects. introduced at the CGIAR institutes, and will The World Bank had an early involvement probably be equally important within the with tidal ponds in Indonesia, the Philippines, medium- to long-term range (10-30 years) Thailand and Nigeria. There has been assis- (Bialy 1992). tance for medium nutrient input systems in The terrestrial orientation of CGIAR Bangladesh and Thailand for shrimp, and in changed when ICLARM in the Philippines China, India, Egypt and Yugoslavia for carp. became a member of the CGIAR network in More recently, the World Bank provided 1992. It was established in 1977, largely 82 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES through a grant from the Rockefeller Founda- were synthesized in a concise World Bank tion. Currently, it is funded at about $4 technical paper published in 1991 (World million per year and is staffed by sixty scien- Bank 1991a). Findings from the study have tists and support personnel. ICLARM's implications that go beyond agriculture. In mission is to assist in the management and particular, the study, among other activities, marketing of aquatic resources in developing analyzed the regulatory and intellectual prop- countries. ICLARM has four basic pro- erty issues as they relate to biotechnology and grams-aquaculture, coastal area manage- developing countries. To sum up the recom- ment, captive fisheries management and mendations, in regard to regulations, the information services. Its beneficiaries are report suggested that developing countries set mainly small-scale fisheries and traditional up institutional biosafety committees in scien- aquaculturists. tific research institutions and establish nation- An example of the type of R&D projects al review bodies and guidelines along the done at ICLARM is the project on the genetic lines formulated by the OECD, to monitor improvement of farmed tilapias (GIFT), and regulate biotechnology research, testing which is being carried out in cooperation with and applications. In regard to intellectual the Institute of Aquaculture Research in property rights, the point is made that the Norway and is funded by the UNDP and lack of patent protection in most developing ADB. The impetus of GIFT came from the countries is a major disincentive for private progressive lowering of yields from existing sector investments in biotechnology, both by tilapia aquaculture due to a deterioration of local private sector companies and by trans- tilapia broodstock in the Philippines, caused national corporations. Each country needs to in part by the introduction of a lower quality weigh the benefits and costs of intellectual tilapia strain from Mozambique (but which property rights in biotechnology, and frame grow well in brackish water). The primary its policies accordingly. The report suggests objective of the project is thus to produce a further that the international agricultural tilapia strain that grows faster than present research centers could also patent their inven- strains and has other favorable characteristics tions, and then license (freely, if appropriate) (Guerrero 1991). these inventions for use by national agricul- Separate from its support of CGIAR, tural research systems (NARS), and other World Bank investments in the agricultural collaborators. research sector for the period 1981-1987 was Outside of agriculture research, some $575 million for twenty-one national projects. other projects have education, science and We cannot determine exactly how much of technology components that include biotech- this money goes to biotechnology, but at least nology activities. Most of this funding has $93 million is provided for biotechnology been directed to more technically advanced R&D in ten different projects. countries. In these cases, funding has been In 1989 the World Bank, in cooperation for infrastructure, laboratory facilities and with the Australian Centre for International equipment, and training. One recent example Agricultural Research, the Australian Interna- is the Support Program for Scientific and tional Development Assistance Bureau and Technological Development (PADCT) in the International Service for National Agri- Brazil. The first part, PADCT I, was com- cultural Research, sponsored a pioneering pleted during 1985-90 and was funded by a study on the opportunities that agricultural $72 million loan from the Bank and Brazilian biotechnology presents for international funds of $107 million. PADCT II will be development. The findings from that study carried out during 1991-98 and will be fund- Major international agencies and blotechnology 83 ed at about $600 million, with funding to be with agency personnel, there seems to be provided equally by the Bank and Brazil. The three major reasons for this situation. First, objectives of this large project is to strength- the professionals staffing agencies are mostly en the management of Brazil's science and unfamiliar with biotechnology generally and technology sector; strengthen Brazilian capa- are almost entirely unaware of marine bio- bilities in specific science areas, including technology. Thus, even in cases where bio- biotechnology; and to improve the milieu for technology could be constructively applied to technological innovation in industry. The solve problems or promote economic devel- biotechnology component, funded at $74.2 opment, they are unable to present the bio- million during PADCT I and $103.1 during technology option to policy makers in devel- PADCT II, aims to strengthen biotechnology oping countries. Second, policy makers in for applications in human health, agriculture, developing countries and their technical animal husbandry and industry. While the advisers, while often aware of biotechnology, major portion of these funds will support are uninformed of marine biotechnology, so scientific research and technology develop- they are not in a position to request assistance ment projects, a significant portion will be in this field. The low level of awareness of used for human resource development, mostly marine biotechnology stems from the third specialized training in-country and overseas. reason, namely marine biotechnology has Lessons from the PADCT project are ex- received little publicity so far. Unlike general pected to be applied to other large science biotechnology, which has been widely and and technology projects. First in line is a massively reported on throughout the world, project that is being developed for Mexico to information about marine biotechnology has completely reorganize that country's main not spread much beyond researchers in the agency supporting science and scientific field and a few industrialists seeking to apply research, the Consejo Nacional de Ciencia y research results, mainly related to natural Tecnologfa (CONACYT). The details of this marine products. Further, the question of program, called Program of Support for what implications general biotechnology have Mexican Science, is being worked out as this for developing countries has been debated by is written. scientists and decision makers in developing countries since 1981-82, when UNIDO Discussion initiated the project that was to become the ICGEB, and when the NAS published its While some agencies described and dis- extremely influential study on biotechnology cussed in this chapter provide technical assis- for development (BOSTID 1982; UNIDO tance in general biotechnology or in marine 1981). No such pivotal events relevant to biotechnology-related areas (such as aquacul- marine biotechnology has yet taken place. ture and natural products development), no Perhaps this report will serve as a starting agency as yet supports projects in marine bio- point for the promotion of marine biotechnol- technology. From the interviews we have had ogy in developing countries. 7 Exploring World Bank options for investments in marine biotechnology In this chapter various options are ex- endemic diseases from damaging the cultured plored for World Bank assistance to develop- animals or plants. Research may be done to ing countries for sustainable development in develop diagnostic methods for early detec- marine biotechnology. Three approaches are tion of diseases, for vaccines to prevent the suggested that are already being taken by the occurrence of diseases, or for therapeutic Bank in other contexts: science and technolo- drugs to treat diseased organisms. The ele- gy lending; support of environmental objec- ments common to Bank projects would have tives; and support for private sector develop- implications for this component: it would take ment (PSD). into account local conditions and would deal with a local problem, it would be an innova- Science and technology lending tive approach to anticipating problems that if not met might negatively affect the project's A former science adviser to the World outcome, and it would probably include the Bank has clarified the four mechanisms transfer of technology and its adaption to fit whereby the Bank supports science and tech- local circumstances. Similar examples could nology (Weiss 1985). Three of these mecha- be given for future projects involving waste nism has implications for capability building water treatment, pollution control and allevia- in marine biotechnology. tion, natural products development, and public health in the coastal zone. Technical assistance for Many opportunities exist for this kind of technological development assistance; we will present one example here. The nation of Qatar now uses revenues from Certain elements are common to Bank its one major natural resource, petroleum, to projects that aim to assist a country in its support its agriculture through expensive technological development. As a rule, Bank imports. In particular, Qatar imports soil officials try to ensure that development pro- nutrients to enrich its sandy soil, hay for jects are well suited for the borrowing coun- feeding live stock, and fish feed for a small try and that they fit existing local conditions. aquaculture industry (Seaweed 1991). How- Many Bank projects include activities for ever, an indigenous alternative could relative- transferring technology to the borrowing ly easily be developed. The nation could country. These elements are all relevant to invest in building land-based tanks or ponds the technological development of the marine for growing macroalgae. With proper R&D sector in borrowing countries. it would be possible to grow macroalgal If a requesting country is one that is rich strains that after processing could be plowed in marine resources, and it therefore could into the soil, adding the vital nutrients and usefully apply marine biotechnology, this body for growing crops. Further, part of the option could be brought to the attention of the harvest may be diverted for use as fish feed borrowing country's officials. For example, in aquaculture. An investment into macro- if the Bank at some future time develops an algal R&D and aquaculture facilities under aquaculture project, it could include a re- circumstances such as these would seem to search component to, for example, prevent make business sense. Exploring World Bank optlons 85 Projea lending for science and technology niques and approaches. For example, marine biotechnology may be introduced into aqua- culture, natural resource exploitation and Some loans are aimed at directly enhanc- other marine-related projects being undertak- ing the scientific and technological capability en in Ecuador, Indonesia, Malaysia, Philip- of a country. The Brazilian Science Research pines and several of the South Pacific island and Training Project is an example of such a countries. In these instances, it would be project; and it might become a model for important that the marine biotechnology be how the Bank may assist biotechnology deployed in such a manner that it fuses with capability building in developing countries. A traditional technologies already in use, en- prospective example may be mentioned. If hancing their scope and possibilities. the World Bank study on fisheries research The specific action that the World Bank that we mentioned above leads to the formu- can take to support science and technology lation of projects for strengthening capabili- for development is to make funding available ties of scientific institutions in fisheries and to governments so they can set up mecha- aquaculture research, these projects might nisms for technology transfer that are similar include biotechnology components to, for to agricultural extension services that are al- example, perform genetic studies on target ready commonly found in developing coun- organisms, clarify the movements of pelagic tries. Specifically, public institutes, whether fish, prevent and diagnose important diseases university or national research laboratory, afflicting target organisms, or improve repro- should be encouraged to set up technology ductive success among culture animals. transfer units according to the terms that are The past history of the World Bank's outlined in Chapter 5. This process can be science and technology lending shows that it enhanced by the World Bank making avail- is primarily directed towards countries with able resources to governments so they can set medium incomes. Marine biotechnology is an up marine extension services. It has funded activity that is most suitable for middle in- similar efforts in the past, especially in agri- come countries in which a considerable culture, so this type of effort should not numbers of people are involved in the biolog- present difficult problems to implement. If ical sciences. In particular, it is clear that the World Bank decides to take up this sug- advanced biotechnology requires a sophisti- gestion, it might find a useful model in the cated research infrastructure and considerable U.S. National Sea Grant College Program long term investment. that is funded by federal funds through the Therefore, in the first instance, science National Oceanic and Atmospheric Adminis- and technology lending to initiate or enhance tration (Ragotzkie 1988). marine biotechnology might be aimed at Sea Grant operates through twenty-nine countries that already have scientific institutes coastal programs, involving hundreds of wherein sophisticated biotechnology R&D is scientists at about forty universities and proceeding, such as Argentina, Brazil, Chile, research institutions. While Sea Grant pro- China, Cuba, India, Mexico and Thailand. In vides an excellent mechanism for universities, the second instance, the Bank could target industry and government to pursue coordinat- countries that have lower level capabilities in ed efforts in R&D, its major function is to the biological sciences, but in which large transfer marine science and technology from marine-related projects are under way that laboratories to technology end-users. It does may benefit from marine biotechnology tech- so by mobilizing support for practically 86 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES oriented research projects, and then acting to potential projects in three ways. First, marine make information about these projects avail- biotechnology techniques may be used to able to industry. The performance of the Sea monitor the possible effects of a project on Grant managers, and indeed of the program the marine environment. This can be done itself, is thus directly tied to its ability to through the use of biosensors, which may be transfer technology, so it is in the agency's designed to detect and monitor specific sub- best interest to make certain practical results stances that may be emitted in the course of flow from the R&D it sponsors. project activities, or molecular biology tech- niques may be used to analyze the chromo- Support to international research somes and/or genes of marine organisms to see if they have suffered damage from project The Bank does not as a rule support activities. scientific research directly; the only excep- Second, molecular biology techniques may tions are its support of CGIAR and WHO's be used to track the movement of pelagic Special Programme for Training and Re- organisms. It is now possible, for example, search in Tropical Diseases. The relationship to determine the origin of a salmon caught on between the World Bank and CGIAR is the high seas through genetic analysis. This described in Chapter 6; the World Bank may will enable regulators to protect salmon find it worthwhile to act so that CGIAR's whose existence is threatened because their purview is extended to include more marine- migratory paths are vulnerable to human related activities. As the situation now stands, predation. Being able to determine which only one of the seventeen IARCs has an country 'owns" salmon, or other pelagic fish, aquatic focus. In view of the possible contri- caught in the high seas may also have impli- bution that the oceans can make to increasing cations for international law. the world's food supply through aquaculture Third, marine biotechnology techniques and of being the source of unique drugs can be used in efforts to clarify the "health" whose use could enhance mankind's health status of coral reefs and mangroves. In par- status, it is important that the World Bank ticular, science now knows little about the encourage IARCs to take up research related juvenile and intermediate forms of inverte- to living aquatic resources and that additional brates and plants that as adults populate reefs IARCs should be established in regions and mangroves. Yet, they are the organisms whose development would be enhanced that are most vulnerable to pollutants or through marine biological and marine bio- physical changes. The origin and fate of these technology R&D. If the latter option were tiny, planktonic forms are difficult to ascer- taken up, it might prove cost effective for tain with present methods. These forms can CGIAR to upgrade existing national facilities be subjected to genetic analysis, using tech- devoted to marine biology, aquaculture or niques such as RFLP and PCR, to clarify fisheries research. speciation. Analysis of their numbers and lineage can lead to a determination of which Support of environmental objectives reef species are diminishing and the reasons why, as well as making it possible for scien- Environmental concerns in some cases tists to clarify certain ecological phenomena dictate whether a particular World Bank such as reef die-offs and plankton blooms. project should be carried out at all, in other The marine biotechnology techniques cases they dictate how projects are imple- related to bioremediation may, of course, be mented. Marine biotechnology may affect used directly to clean up polluted marine Exploring World Bank optfons 87 environs, but we consider this possibility in research sector. To illustrate, say that the the next section. World Bank considers a project to establish Relating directly to environmental initia- an automobile manufacturing plant in a devel- tives, there is at this time much discussion oping country. Although many of the technol- about criteria for lending of the newly estab- ogies required in this endeavor will be com- lished GEF. It is clear that at least two of the plicated and some may be designated as high four sectors of the GEF will include activities technologies, the implementation of this where marine biotechnology could be compo- project could be done wholly without the nents. One is international waterways and involvement of universities or their research- marine environments and the other is conser- ers because the technologies are known, their vation of biological diversity. On improving implementation requires engineers and techni- marine environments we have already men- cians, and their operation will be the same tioned the reception facilities for wastes that regardless of where it is done. the GEF is committed to constructing. Assist- The situation with biotechnology-based ing cleanup of accidental spills of oil could be industry is different because it is science- another activity. Monitoring of changes in based. In this case, the molecular control marine environments could draw on the over the metabolism and reproduction of improvements of monitoring technologies that industrial organisms must be employed for marine biotechnology has brought about. applied purposes. This implies that the natu- Marine biodiversity is also covered by the ral phenomena underlying these processes are GEF. There should be possibilities in demon- sufficiently well known so that they can be strating the potentials for using biological manipulated for preconceived ends. Only diversity in the seas for developing new well-trained scientists can do so. Further, products. An illustrative project that would many times when scientists elucidate a natural show the value that biological diversity in the phenomenon, they simultaneously generate seas have for commercializing sea products the knowledge that can drive an industrial might improve the interest of developing process. For example, when a researcher countries to maintaining their living marine locates and characterizes a gene in an organ- resources. ism that codes for the production of a specific protein, he or she in the same instance dis- Support for private sector development covers a process that can be used by industry to manufacture that protein in vitro. Since Supporting PSD has become a major this is the case, a biotechnology-based indus- World Bank goal because it fulfills the Bank's try cannot be established in a country unless primary objectives of reducing poverty and there is a close relationships between the raising standards of living. The four types of research establishment and the applied sector. activities the Bank uses to promote PSD are Referring to the concept development described in the World Bank's 1991 Annual process discussed above, when considering Report. Most are generic; that is, their ac- projects involving biotechnology, research complishments should lead to PSD whether it laboratories should be linked with technology is in agriculture, food industry, chemical end-users in industry, agriculture and health. industry or biotechnology-based industry. But As is discussed above, this has to be done one important point needs to be made con- from two directions. The first direction, that cerning PSD related to biotechnology, namely of universities and national research institutes that biotechnology-based industry, wherever reaching out to technology end-users, possi- it is to grow, must be closely tied to the bly through a Sea Grant-like approach, has 88 MARINESIOTECHNOLOGYAND DEVELOPING COUNTRIES been addressed. The second direction is the Table 1. Sectors appropriate for marine technology end-user accessing results from biotechnology research done at public institutes. The World Bank, as part of national programs to assist Technology and tbneirwnu indigenous industry, could make funding available for firms to establish advanced uctor Shorterm Long tam research and development units capable of Industry Bioremediation Sensors adopting and adapting research results. This Agriculture, Genetic Specific might be done by setting up a special funding Aquaculture Modification Proteins agency within federal governments in, for Health Diagnostics Pbhzmazical example, ministries of commerce or industry. Transportation Bioremediation Biofilms The intent of this new agency would be to Sanitation Bioremediation Sensors provide loans at favorable rates to small and medium businesses so they can hire the scientists and buy the equipment to perform other biotechnology relates to the industry the advanced research and pilot trials for new that specializes in pollution abatement and product development. Of course, this type of control. One of the sectors of that industry is initiative would have to be integrated with remediation. The remediation industry is one other efforts to improve the business climate of the fastest growing industries in the world; in a country, including business tax reform, at present, its market is estimated to be $2 adoption of intellectual property laws and billion per year and its annual growth rate for safety regulations, and the possibility of firms the next five years is 25 percent (Chowdhury being able to import required technology and 1992). Bioremediation is one of the tech- supplies without undue hindrance. An exam- niques used by this industry, usually when it ple of how this can be done is provided by involves treating polluted coasts and water- the Brazilian PADCT project discussed ways. above, which includes a component that aims The remediation industry is now concen- to improve on the ability of industry to inno- trated in industrial countries. But it is certain vate. The setting up of an advanced research to spread to developing countries, especially and development unit by a firm should ac- so since the technology is available, it is complish this objective. relatively cheap, there are many opportunities The other PSD activity that is relevant to for local research to improve bioremediating the promotion of biotechnology is its direct organisms and techniques, and there are fostering of private enterprise, including many situations where bioremediation can be providing support for entrepreneurial efforts employed immediately for great effect. (World Bank 1992). The Bank is involved in Another PSD-type project could be aimed many sectors where there are possibilities for at establishing regional-based firms to raise adding value to raw material products via and maintain broodstock and to enhance seed biotechnology. This would increase economic production for aquaculture industry. This is return to the developing country producing one of the major bottlenecks to aquaculture. the raw material and help it lessen its techno- For example, India's shrimp aquaculture logical dependence on outsiders. Biotechnolo- industry requires about 2 billion fry per year gy could thus be suitable for several sectors and the industry's demand increases 10 per- (see Table 1). cent every year (Kant 1991). Yet, only 17 An initiative that has environmental impli- hatcheries are in operation, each producing cations and that could employ marine and fewer than 40 million fry. Aquaculturists Exploring World Bank optlons 89 make up the difference by collecting from the potential to support projects that might seem wild, which is environmentally damaging. too risky for political or commercial reasons This situation is not unique; the major con- in developing countries. Capitalizing on the straint on mollusc aquaculture in Asia is the stability that IFC provides in the development lack of seed (Lovatelli 1990). Making avail- of new commercial projects, and considering able funding for setting up hatcheries, seed the smaller projects that it can support, the production units, and facilities for maintain- IFC could support the development of marine ing broodstock would be environmentally biological joint ventures. Before it can do so, kind and good business. however, the IFC would have to develop a The final point regarding PSD, the Inter- stronger technical competence in the field so national Finance Corporation (IFC), which is that it can properly assess the feasibility of the member of the World Bank group that biotechnology projects, thus minimizing the supplies direct project financing for private risk of initial failures. investments in developing countries, has the 8 Conclusion When exploring options for marine bio- important, however, that henceforth the technology for developing countries, it be- World Bank must take its potentials into came clear that we must appraise a larger account. issue first. To wit, what is appropriate assis- Developing this view further, it is realistic tance to developing countries for biotech- to believe that in the future, as in the past, nology programs or projects? We have the Bank will be called on to provide assis- shown in Chapter 2 and elsewhere in this tance to countries to improve their agricul- report that of all the high technologies, bio- ture. In these cases, plant biotechnology technology is the most appropriate for devel- could well become an element in projects to, oping countries because: entry into the field for example, increase the resistance of plants is easier and costs less than any of the other to diseases or pests, increase the ability of high technologies; many developing countries crops to grow under arid conditions, design have an existing base in the natural sciences crops to tolerate brackish water, or for other from which biotechnology can develop and purposes that helps the particular country in grow; many developing countries have rich question. In other cases biotechnology may natural resources that can be exploited via be used in projects having health objectives biotechnology for sustainable, environmental- to, for example, to improve drugs or develop ly sound economic development; and certain vaccines; to expand industrial capabilities by, problems facing populations in developing for example, utilizing agricultural wastes for countries related to disease, food supply, alcohol production; and so forth. Similarly, environmental degradation and energy supply projects aimed at assisting island countries or may be amenable to technical solutions that countries with significant marine resources only biotechnology can provide. In recogni- may include marine biotechnology compo- tion of the promises that biotechnology holds, nents for objectives such as increasing yields scientists and political leaders of developing from aquaculture, marine natural products countries have already requested assistance development, the improvement of public from many public and private organizations, health, or to clean polluted coastlines. The who have responded by sponsoring activities point of the foregoing is that a Bank officer, designed to help enhance present capabilities when planning wide-ranging projects on land in biotechnology and acquire new ones. or related to the sea, whether in the agricul- The World Bank has not been one of these ture, health, industry or environment sector, organizations, except in two cases: in large may want to consider the possible benefits projects in Brazil and Indonesia that have that the application of biotechnology tech- sizeable biotechnology components, and in its niques may have for reaching project objec- support of CGIAR that funds some aquacul- tives, something that probably has not been ture and biotechnology R&D. The field has done so often in the past. grown to such an extent and has become so Appendix A Marine biotechnology and related R&D institutions in developing countries Short questionnaires were sent to a select Ciencias da Vida (Dr. Celina Roitman), number of scientists in developing countries Conselho Nacional de Desenvolvimento who had been identified as being involved Cientifico e Tecnoldgico. Strategic with marine biotechnology or biotechnology- planning for the utilization of marine related areas. The answers to these resources is done by the Comisslo questionnaires, suitably organized, are set Interminesterial de Recursos de Mar forth in the sections that follow. Thus, each (CIRM); plans are implemented by the section is devoted to one country. It begins Secretaria Ciencia e Tecnologia (SCT) by naming the ministry or other authority (Mr. Paulo Cesar Goncalves Egler). under whose purview marine biotechnology falls. If possible, the person in charge is B. R&D Institutions. named. Thereafter the country's research institutes performing marine biotechnology, 1. Universidade Federal do Maranhao or related R&D, are listed. When known, the (UFMA), Fortaleza. R&D areas of institutes are specified. a. LABOMAR: marine biology. The information should be used with 2. Universidade Federal Rural de caution for two reasons. First, since marine Perambuco, Dois Irmaos. biotechnology, and its related areas, is a. Departemento de Engenharia de rapidly expanding, the work programs of Pesca. research institutes are also growing and b. Departemento de Biologia. changing. As a result, new departments are 3. Universidade Federal de Alagoas, being set up, old departments expanded, and Macei6: marine biology. additional scientists are being hired to staff 4. Universidade Federal de Sergipe, them. Administrative changes may have been Aracaju. instituted at local and national levels that a. Departemento de Biologia: mangrove reflect the growing importance of marine biology. biotechnology. For these reasons, the 5. FIPERJ, Rio de Janeiro: aquaculture. information provided here should be 6. Universidade do Estado do Rio de considered a 'snapshot" of a rapidly Janeiro, Rio de Janeiro. developing and changing field, one that a. Departemento de Oceanografia: depicts the situation as it existed in late 1991. marine biology, marine pollution. Second, we were entirely dependent on 7. Instituto de Estudos do Mar Almirante information supplied by persons who head or Paulo Moreira, Arraial do Cabo: work at the institutions that are listed. Some macroalgae. of this information may represent wishes or 8. Universidade de Sgo Paulo, Sgo Paulo. aspirations rather than present reality. a. Departemento de Zoologia: shrimp diagnostics. Brazil. a. Instituto de Biociencias: shrimp baculovirus, macroalgae. A. The agency responsible for marine- b. Instituto Oceanografico: marine related R&D is the Coordenadoria de biology, marine pollution, ichtyology. 92 MARINE A90 TECHNOL OGY AND DEVELOPING COUNTRIES 9. Universidade Federal do Parana, Abeliuk) and the Comit6 Oceanografico Paranagud. Nacional (Mr. Hugo Gorziglia Antolini). a. Centro de Biologia Marina: marine In addition, Chile has a national biology, plankton studies, bacteriobentos biotechnology program that is guided by in mangroves. the Comit6 Nacional de Biotecnologfa 10. Universidade Federal do Rio Grande do (Dr. Jorge Allende). Sul, Porto Alegre. a. CECO: marine ecology. B. R&D Institutions. 11. Fundacao Universidade do Rio Grande, Rio Grande. 1. Universidad Cat6lica de Valparaiso, a. Departemento de Biologia: Valparaiso. phytoplankton studies. a. Instituto de Biologfa: cloning of b. Departemento de Biologia e genes from marine microorganisms; Anatomia: fish pathology. reproduction of marine organisms; c. Departemento do Oceanografia: ecology of aquatic populations. marine biology. b. Escuela de Ciencias del Mar: d. Laborat6rio de Fitoplancton: marine pathology of salmonids; evaluation of biology. fish feed; reproductive control in fish; 12. Universidade Federal da Bahia, Bahia. production of triploid salmonids; algal a. Instituto de Biologia: algae bank, and mollusc aquaculture; marine bioassay of water quality, shrimp and pollution research. oyster aquaculture. c. Escuela de Alimentos: study of 13. Universidade Federale do Santa Catarina, protein structure and enzymes. Floriandpolis. d. Escuela de Ingenierfa Bioqufmica: a. Departemento de Aquicultura: shrimp utilization of microalgae; pilot plant aquaculture, reproductive fish and shrimp production of algal products, including technologies. pigments, proteins and lipids. b. Departemento de Biologia: bioassay e. Escuela de Ingenerfa Qufmica: lipid of marine waters, antioxidant protection extraction. of marine vertebrates, crustacean f. Laboratory at Vifia: macroalgae immunology, bacterial accumulation by tissue culture. mussels and oysters. 2. Universidad de Magallanes, Punta c. Departemento de Bioquimica: sea Arenas. anemone toxins. a. Instituto de la Patagonia: d. Departemento de Zooligia: shrimp environmental protection. pathology. 3. Instituto de Fomento Pesquero, Santiago (Dr. Patricio Bernal Ponce). Chile. a. Laboratory at Putemdn (in cooperation with the Universidad Cat6lica de Chile): A. The authority having the main study and management of marine responsibility for marine biotechnology resources, including mollusks and algae; or related areas is the Subsecretarfa de induction o f mussel larvae Pesca (Mr. Andrds Couve) of the metamorphosis. Minesterio de Planificacion (Mr. Sergio b. Laboratory at Coyhaique: aquaculture Molina). Other agencies having pertaining to salmonids. responsibilities in these fields are the 4. Universidad Cat6lica de Chile, Santiago: Corporacion de Fomento (Mr. Rend studies on macroalgae and pigment Appendix A 93 production. B. R&D Institutions. 5. Universidad Catolica de Chile, Sede Talcahuano, Talcahuano: macroalgae 1. Marine Science Laboratory, Department genetics. of Biology, Chinese University of Hong 6. UniversidaddeConcepcidn, Concepcidn: Kong: fish and shrimp mariculture. studies on microalgae and pigment production. India. 7. Universidad de Santiago, Santiago: studies on crab and pigment production. A. Major responsibility for biotechnology in 8. Lefersa Alimentos, Santiago: studies on India rests with the Department of feed for salmonids. Biotechnology (Dr. C. R. Bhatya) of the Department of Science and Technology, Egypt. Ministry of Science and Technology. Responsibility for marine affairs lies with A. The Ministry of Agriculture has the Department of Ocean Development responsibility for fresh water fisheries (Dr. S.N. Dwivedi). The Marine and aquaculture, while the National Products Export Development Authority Institute of Marine Sciedces and (Mr. Amitabh Kant), Ministry of Fisheries, Academy of Scientific Commerce, has responsibility for the Research and Technology, Ministry of development of India's seafood industry, Scientific Research is responsible for including export production and marine living resources and related promotion. R&D. B. R&D Institutions. B. R&D Institutes. 1. National Institute of Oceanography, 1. National Institute for Marine Sciences Dona Paula, Goa. and Fisheries, Cairo. a. Division of Microbiology: hydrolytic 2. Egyptian Authority for Fisheries and enzymes from marine bacteria; Fishing Gear, Alexandria. microalgal production of beta carotene, 3. Ministry of Agriculture, Cairo. glycerol and proteins; screening of a. Agricultural Development Project of marine microorganisms for bioactive Eastern Abbassa. compounds; shrimp aquaculture; algal b. General Authority for Fish Resources tissue culture. Development. b. Marine Corrosion and Materials c. Maryoot Project for Aquaculture. Research Division: extracellular 4. Suez Canal University, El-Arish. production by marine microorganisms; a. Faculty of Environmental Agriculture. bioremediation. 2. Goa University, Taleigo, Goa. Hong Kong. a. Department of Marine Biotechnology. b. Department of Marine Sciences (in A. Marine biotechnology appears to fall Bambolim): marine biology, marine under the purview of the Agriculture and microorganisms. Fisheries Department, Hong Kong 3. Central Marine Research Institute, Government. Cochin, Kerala. 4. Sri Paramakalyani College, Kallidaikurichi. 94 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES a. Post Graduate Department of B. R&D Institutions. Microbiology: screening of marine bacteria, algae and invertebrates for 1. School of Biological Sciences, University bioactive compounds. of Malaysia, Penang. 5. Centre for Advanced Study in Marine 2. Faculty of Biology, National University Biology, Porto Nova. of Malaysia, Selangor. 6. Anna University, Guindy, Madras. a. Centre for Water Resources. Mexico. 7. Cochin University of Science and Technology, Kochi. A. The authority responsible for all science a. School of Marine Sciences: marine and technology in Mexico is the Consejo bacterial enzymes as find chemicals; Nacional de Ciencia y Tecnologfa immobilized enzyme systems for water (CONACYT), Mexico, D.F. quality control; marine microbial enzymes; fish feed formulation; algal B. R&D Institutions. cultivation; pollution toxicology. 8. Calcutta University, Calcutta, West 1. Centro de Investigaci6n Cientffica y de Bengal. Educaci6n Superior de Ensenada, Baja a. Department of Marine Sciences: California. intertidal ecology; ecology of mangroves. a. Laboratorio de Biotecnologfa Marina: 9. Beshampur University, Beshampur, fish vaccines; detection of Vibrio species; Orissa. pollution control; fermentation by marine a. Department of Marine Sciences: bacteria. chemistry of the sea. 2. Centro de Investigaciones Biol6gicas de 10. Andrah University, Waltair, Andrah Baja California Sur, La Paz, Baja Pradesh. California. a. Department of Marine Sciences: 3. UABCS Departemento de Biologfa marine biology. Marina, La Paz, Baja California. 11. Kerala University, Trivandrum. 4. E.N.B.C. - I.P.N., Santo Tomgs, a. Department of Aquatic Biology and Mexico D.F. Fisheries: marine biology; marine 5. Departemento de Biotecnologfa, Instituto microbial enzymes; bioactive compounds de Investigaciones Biomedicas - UNAM, from marine organisms; marine ecology. Mexico D.F. 12. Karnataka University, Karwar, 6. Secci6n de Biotecnologfa, Instituto Karnataka. Tecnologico de Merida, Merida, a. Department of Marine Biology: Yucatan. marine biology. 13. Government Institute of Science, Nigeria. Aurangabad. a. Department of Microbiology: A. The authority responsible for all science halophilic microorganisms; andtechnologyistheFederalMinistryof bioremediation. Science and Technology (Dr. G. Ezekwe). Malaysia. B. R&D Institutions. A. Authority to be determined. Appendix A 95 1. University of Port Harcourt, Port bioadhesion; biofouling; natural Harcourt. chemicals from marine bacteria; studies a. Department of Microbiology: on human pathogens in coastal waters; bioremediation; the isolation of studies on infectious diseases afflicting biopolymers from marine bacteria and prawn. algae. b. Marine Genetics Laboratory, 2. Nigerian Institute for Marine Biology Department of Marine Biology: genetics and Oceanography, Lagos: fisheries and of prawns and macroalgae. oceanography. 2. Institute of Oceanology, Academia 3. Imo State University, Okigwe, Imo Sinica, Qingdao, Shandong. State. a. Marine Microbiology Laboratory: studies on diseases afflicting the People's Republic of China. aquaculture of prawns and macroalgae. b. Seaweeds Laboratory: genetics of A. There appears to be an overlap of macroalgae; development of productive responsibilities for marine biotechnology techniques for macroalgae. in China.The Administration for Science c. Invertebrate Laboratory: scallop and Technology (Mr. Song Chian) seems aquaculture; prawn aquaculture; to have major responsibility for scientific biofouling. research, while the Scientific and 3. Experimental Marine Biology Technological Section (Dr. W.H. Yang) Laboratory, Academia Sinica, Qingdao, of the State Oceanic Administration is Shandong: research on transgenic fish; responsible for marine-related research. the construction of a fish gene library; Further, the Department of Earth photosynthesis of algae; applications for Sciences (Dr. Yang Sheng) of the Bureau Spirulina; the immobilization of algae of Earth Sciences and the Bureau of and the use of algae in bioreactors; algae Aquatic Products of the Ministry of tissue culture; in vitro fertilization of National Agriculture, Animal Husbandry shrimp and scallops; transgenic scallops; and Fishery also have interest in marine and manipulation of reproductive biotechnology. In addition, the Marine processes in shrimp and scallops. Sciences Section (Dr. Y.B. Fan) of the 4. Yellow Sea Fisheries Research Institute, National Natural Science Foundation of Qingdao, Shandong: algae and prawn China and the Bureau of Bioscience and aquaculture; study of diseases afflicting Biotechnology (Dr. G.Z. Meng) of the aquaculture; technologies of fisheries Chinese Academy of Sciences are products. involved with biotechnology activities. 5. First Institute of Oceanography, The delineation of authority between Qingdao, Shandong: marine ecology; these agencies vis-a-vis marine prevention of fish diseases. biotechnology remains to be clarified. 6. Second Institute of Oceanography, Hang Zhou: biochemistry of marine B. R&D Institutions. organisms. 7. Third Institute of Oceanography, 1. Ocean University of Qingdao, Qingdao, Xiamen: expression of fish growth Shandong. hormone in E. coli; biochemical a. Marine Microbiology Laboratory, monitoring of marine pollutants; Department of Marine Biology: utilization of natural products from 96 MARINE B0OTECHNOLOGYAND DEVELOING COUNTRIES oysters, horseshoe crab, and others; pollution in ocean, macroalgac accumulation of heavy metals by fish. taxonomy. 8. Shanghai Fisheries University, Shanghai: 6. Universidad Nacional de Trujillo: fish studies on macroalgae. biology, invertebrate ecology. 9. South China Sea Institute of Oceanology: 7. Universidad Nacional de Chiclayo research on SplWlna. Pedro Ruiz Gallo": sand beach 10. Institute of Genetics, Academia Sinica, ecology. Beijing: antifreeze protein from P. 8. Universidad Nacional de Arequipa: yokohamae. marine resources evaluation. 11. Institute of Hydrobiology, Academia 9. Centro de Investigacfon de Bioqufmica y Sinica: nitrogen fixation in cyanobacter. Nutricfon. 10. Instituto Tecnol6gico Pesquero: marine Peru. natural products, processing of marine living resources. A. The agency that is responsible for science and technology is the Consejo Philippines. Nacional de Ciencia y Tecnologfa, while the Fondo de Reactivacfon del Sector A. The agency having responsibility over Pesquero (Ing. Luis Garayar Melendez) science and technology is the Department of the Ministerio de Pesquerfa (Dr. Fdlix of Science and Technology (Mr. Canal Torres) has responsibility over Ceferino Follosco). The Bureau of ocean resources, including aquaculture. Fisheries and Aquatic Resources, Department of Agriculture, has as its B. R&D Institutions. name suggests authority over aquatic resources. 1. Instituto del Mar del Peru, Callao: This institute is responsible for R&D B. R&D Institutions. pertaining to fisheries, aquaculture, and other marine activities. It has four 1. University of the Philippines at Los regional, coastal laboratories at Banos. Chimbote, Ilo, Paita and Pisco, as well a. Learning Resource Center: studies on as three research vessels. fresh-water algae as a source of 2. Universidad Nacional Jorge Basadre carrageenan. Grohman, Casilla. b. Marine Science Institute: genetic 3. Universidad Nacional Faustino Sanchez studies on Euchewma, culture of Carri6n de Huacho, San Isidro. Gelidiella, biology and ecology of 4. Universidad Nacional Mayor de San Saragassum. Marcos, Casilla. c. Seaweed Information Center (in a. Biological Sciences Faculty: Quezon City), Marine Science Institute: macroalgae aquaculture, mollusc studies on macroalgae. aquaculture, plankton upwelling, 2. College of Fisheries, University of the reproduction in fishes and mollusks. Philippines in the Visayas: seaweed 5. Universidad Federico Villareal, processing and utilization. Miraflores. 3. College of Fisheries, Bicol University: a. Facultad de Oceanograffa, Pesquerfa seaweed processing and utilization. y Ciencias Alimentarias: heavy metals 4. Marine Biological Laboratory, Silliman Appendx A 97 University, Dumaguete City: research Taiwan (China). on seaweed species. 5. University of San Carlos, Cebu: A. Fundamental research in biotechnology is research on seaweed species. the responsibility of the Life Sciences 6. Aquaculture Department of the Southeast Division (Dr. Jung-Yaw Lin), National Asian Fisheries Development Center, Science Council, while the Iloilo: research on Gracilaria biology, implementation of the National culture and processing. Development Plan on Marine Science 7. Fisheries Resources Research Division, and Technology, 1991 - 1995, which Bureau of Fisheries and Aquatic includes marine biotechnology, falls Resources: field research on under the purview of Fisheries macroalgae. Department (Dr. Jen-Chyuan Lee), S. Marine Colloids, Inc., Cebu City: Council of Agriculture. commercial production of carrageenan from the alga Eucheuma. B. R&D Institutions. 9. FMC Corporation: research on commercially valuable macroalgae. 1. Academia Sinica, Taipei. a. Institute of Chemistry: seaweed Republic of Korea. polysaccharides. b. Institute of Zoology: fish hormonal A. The Office of Fisheries (Mr. Seong- regulation, transgenic abalone, transgenic Hwan Hah), Fisheries Research and fresh and saltwater fish, IPN virus Development Agency, is in charge of studies, gene expression of RNA virus in fisheries R&D. fish, diagnostic kits for eel and shrimp viral diseases. B. R&D Institutions. 2. Fisheries Research Institute, Tainan: field experiments of fish treated with 1. National Fisheries University of Pusan, hormones. Pusan. 3. Fisheries Research Institute, Penhu: a. Department of Aquaculture. seaweed stock improvement. b. Department of Microbiology. 4. Kaohsiung Medical School. c. Department of Biological Science and a. Department of Microbiology: studies Technology. on gonyautoxins. d. Department of Fish Pathology. b. Department of Pharmacy: natural e. Department of Marine Biology. products from soft corals, monoclonal f. Department of Applied Chemistry. antibodies for gonyautoxins. g. Institute of Marine Sciences. 5. National Chung Hsing University. h. Institute of Fishing Science and a. Institute of Botany: proteins from Technology. seaweed. i. Institute of Sea Food Science. b. Institute of Soil: marine bacteria in j. Institute of Sea Culture. pond soil. k. Institute of Life Science and 6. National Defense Medical College. Biotechnology. a. Department of Microbiology and 1. Environmental Research Institute. Immunology: marine actinomycetes 2. Korean Ocean Research and exploitation. Development Institute, Seoul. 7. National Pingtung Agriculture College: 98 MARINE BIOTECHNOLOGYAND DEVELOPING COUNTRIES fish lymphocystic viral studies. 14. Tong Wu University. 8. National Sun Yat-sen University. a. Department of Microbiology: natural a. Department of Biology: luminescent products from marine bacteria. bacteria as biosensors. 15. Yang Ming Medical School. b. Department of Marine Resources: a. Department of Biochemistry: natural zoology of corals, natural products from products from marine bacteria. coral-associated bacteria. 9. National Taiwan Normal University. Thailand. a. Department of Biology: fish hormone studies. A. Responsibility for marine biotechnology 10. National Taiwan Ocean University. appears to be split between two agencies. a. Department of Aquaculture: seaweed On the one hand, responsibility for the protoplasts, marine bacteria in national biotechnology plan rests with the aquaculture, enzyme inhibitors from National Center for Genetic Engineering marine bacteria, transgenic prawn. and Biotechnology (Dr. Bhumiratana), b. Department of Food Science: marine Science and Technology Development toxins, natural products from bacteria. Agency. On the other, marine resources 11. National Taiwan University. fall under the authority of the a. Department of Agricultural Department of Fisheries (Dr. P. Chemistry: bioremediation, waste water Surasvadee), Ministry of Agriculture and treatment with halophytes. Cooperatives. b. Department of Botany: detection kits for Vibrio anguillarum. B. R&D Institutions. c. Department of Zoology: fish hormone studies, IPN viral studies, eel 1. Department of Microbiology, King herpes viral studies, eel disease Mongdut's Institute of Technology, diagnostic kits. Thonburi: improvement of bacterial d. Institute of Biological Chemistry: strains for fermentation of fish sauce. studies on fish hormones. 2. Institute of Marine Science, Burapha e. Institute of Fisheries Science: fish University, Chonburi: screening of hormone studies, seaweed natural marine bacteria, phytoplankton and products, eel vaccines against zooplankton for biologically active Edwardsiella and Aeromonas, vaccine for substances; the study of heavy metal shrimp vibriosis, studies on adjuvants for resistance in microalgae. shrimp vaccine. 3. Department of Marine Science, f. Institute of Oceanography: seaweed Chulalongkorn University, Bangkok: biology, biology of soft corals, marine improvement of aquaculture practices. thermotropic bacteria utilization. 4. Aquatic Resource Research Institute: 12. National Tsing Hua University. management of aquatic resources in fresh a. Department of Chemistry: natural and salt water, coastal zone resource products from seaweeds. management. b. Institute of Life Science: exotoxin 5. Srinakharinwirot University, Prasarnmit from Edwardsiella. Campus, Bangkok: selection of agar- 13. Taiwan Provincial Research Institute for bearing macroalgae for aquaculture; Animal Health: eel vaccines against improving agar production of the alga Edwardsiella and Aeromonas. Gracilaria. Appendix A 99 6. Brackishwater Fisheries Division, Uruguay. Kasetsart University, Bangkok: mariculture of black tiger shrimp. A. The ministry most concerned with 7. Faculty of Natural Resources, Prince of marine biotechnology will be determined. Songkla University, Haadyai. 8. Marine Biotechnology Laboratories, a B. R&D Institutions. specialized laboratory of the National Center for Genetic Engineering and 1. Instituto de Investigaciones Pesqueras, Biotechnology, located at the Universidad de la Repilblica, Chulalongkorn University, Bangkok: Montevideo: animal feed from fish; the aquatic animal health, marine natural use of proteolytic marine yeast to products development. produce fish protein concentrate. 9. Marine Biological and Fishery Research Institute, Department of Fishery, Phuket: aquaculture and marine natural products development. Appendix B Special equipment requirements for advanced biotechnology Equipment requirements for biotechnology Table B.1. Basic equipment for R&D include optical microscopes, dissecting biotechnology R&D scopes, centrifuges, incubators, sterilizers, tube and flask shakers, glass and plastic lio Apprvbatw price ware, chemicals, media, controlled lights, and growth chambers and greenhouses. The Computer for sequence analysis S 11,000 more specialized marine biotechnology CO2 incubator 4,000 research requires aquaria (i.e., open ponds Digitizer 10,000 and closed tanks) together with a seawater Electrophoresis separation equipment 10,000 system for supporting it; and shops for Froezer (to -70° C) 6,000 mainWnng, rpairig and,as ned be, High-speed refrigerated centrifuge 22,000 coaintaining, repairing and, as need be, Invertod phase microscope 10,000 constructing this specialized equipment. Some Liquid air pump with a 180-liter research in marine biotechnology requires storage tank 18,000 siting the laboratory close to the ocean; other Reserch-grade double-beam work (e.g., natural marine products R&D) is Vpectrophotometer 28,000 not site dependent. All biotechnology R&D Scanning densitometer 18,000 institutions should provide strong support Scintillation counter 30,000-35,000 facilities, such as libraries, electronic shops, Ultracentrifuge 50,000 technical equipment service, information and computer services. The material requirements for rDNA technology are significant, as Tables B. 1 and B.2 demonstrate. Items listed in Table B. 1 Their reagent requirements are also prodi- are those required by a laboratory wishing to gious; for example, in the United States a take up plant biotechnology research; the heavily used automated DNA sequencer can equipment demands for biomedical research consume about $40,000 worth of expendable would be greater. Those found in Table B.2 supplies per year. are needed to perform exceedingly Monoclonal antibody work will require sophisticated research; in developing facilities for culturing animal cells, an animal countries some items may be found only in a house, appropriate animals, and the services national or regional research institute. When of one or more animal handlers. examining the two tables, keep in mind that The chemical and special substances needs equipment upkeep requires a service unit of a biotechnology laboratory are manned by skilled maintenance and repair considerable, including radioisotopes, sera, technicians and possessing an adequate antisera, enzymes, restriction enzymes, inventory of spare parts. Without such media, buffers, antibiotics, and others. In the backup, the equipment will soon break down United States, a researcher or technician and stay down. Further, each piece of requires chemicals and reagents whose costs equipment listed in Table B.2 requires the range from $5,000 to $10,000 per year. services or a specially trained technician. Costs of these expendable supplies would un- Appen*5 101 doubtedly be higher in developing countries Table B.2. Highly sophisticated equipment as most must be imported. Further, since for biotechnology R&D many of these reagents deteriorate rapidly at ambient temperature, suppliers must take emn Approxlm2e price elaborate measures to make certain that they are properly handled in shipping and Automated DNA sequencer $100,000 storage. Flow cytometer 160,000 Equipment and supplies, while of lesser Oligonucleotide synthesizer importance than highly trained scientific (manual model) 11,000 personnel, are essential ingredients to the Oigonucltotide synthesizer carrying out of research; without them some (automabd sysbm) 48,000 projects must be flregone, others will be curtailed. Equipment limitations could also prevent the development of advanced capabilities in biotechnology. Appendix C Definitions of marine biotechnology by scientists in industrial and developing countries "Marine biotechnology is the integration be called biotechnology, so one can apply this of advances in marine microbiology, marine definition to marine biotechnology." (Dr. biochemistry (including cell biology, M.S. Andhale, Department of Microbiology, molecular biology and molecular genetics), Government Institute of Science, marine biology and process engineering, for Nipatniranjan, A'bad Caves Road, application in such areas as food and feed Aurangabad 431 004, India). industry, pharmaceutical industry, environmental pollution and energy, medical "Marine biotechnology is a branch of diagnostics, fermentation industry, and marine science dealing with marine organisms chemical industry." (Dr. Gideon Abu, to enhance the production of food, feed and Department of Microbiology, Box 274, chemicals for the betterment of mankind." University of Port Harcourt, Port Harcourt, (Dr. N.B. Bhosle, National Institute of Nigeria). Oceanography, Dona Paula, Goa-403 004, India). "The term biotechnology generally implies the application of technology to organisms. In "Given the fact that biotechnology is any other words, we try to mould the organisms aspect of biological system that makes or its function to achieve our target. money, I would say that marine Nonetheless, to say better exploration and biotechnology is any aspect of biotechnology exploitation of the ocean and the organisms that either directly concerns aquatic (marine there in for the transmogrification of and freshwater) systems or had as its origin mankind. We would like to define marine an aquatic biological system." (Dr. Joseph biotechnology as the application of genetic Bonavenatura, Director Marine Biomedical engineering to marine sciences i.e. to utilize Center, Duke University Marine Laboratory, the untapped gene pool in: North Carolina). 1. The transport of minerals (nutrient cycle) 2. Novel photosynthetic system (primary "Marine biotechnology can be defined as production) the efficient utilization of marine living 3. Utilization of H2S, NH3, H2 etc resources or their components to provide (chemosynthesis) desirable products and services." (Dr. M. 4. Production of fish, mollusks, crustaceans Chandrasekaran, Microbiology Laboratory, in natural and hatchery system (secondary Department of Applied Chemistry, Cochin and tertiary production) University of Science and Technology, Kochi 5. Marine pheromones, toxins, and 682022, India). pharmacological compounds." (Dr. Shanta Achuthankutty, National Institute of "The application of biological sciences Oceanography, Dona Paula, Goa 403 004, which utilizes living marine organisms, their India). cells or parts of cells to produce good and services." (Dr. S.T. Chang, Department of "I consider that any proven technology, Biology, The Chinese University of Hong which is aided by the biological systems, can Kong, Shatin, N.T., Hong Kong). Appendlx C 103 'Marine biotechnology, an extension of "I would like to define marine marine biology, blends science and biotechnology from my understanding that: technology to develop the methods for mass any marine biological knowledge which could production and processing of marine be applied to increase yield or marine organisms for a wide range of industrial and products is marine biotechnology. Marine commercial uses." (Dr. Saipin Chaiyanan, biotechnology is very wide in the sense, there Department of Microbiology, Faculty of are a lot of things to be done in the field of Science, King Mongkut's Institute of marine biotechnology. For example, only Technology Thonburi, Bangmod, Rasburana marine bacteria and marine plankton can play Bangkok 10140, Thailand). very important role in marine biotechnology." (Dr. Twee Hormchong, "In my opinion, the task of biotechnology Director Institute of Marine Science, Burpha is to synthesize the modem theory and University, Bangsaen, Chonburi 20131, methods of engineering and biology, to Thailand). research the variations of biological structure and function on different level and artificially "I would say that marine biotechnology is to control these variations by using the use of all the tools and knowledge in the engineering and technique, in order to life sciences to produce a desired effect on or develop some new types of industry or new for mankind." (Dr. Robert S. Jones, biological products on a large scale, such as Director Marine Science Institute, University genetic engineering, cell engineering, enzyme of Texas at Austin, Texas). engineering, microbial engineering, biochemical engineering and the technique of "I define marine biotechnology as: The comprehensive utilization for biological use of marine organisms or their genetic resources. ' (Dr. Chen Dou, Institute of information, for applications on aquaculture, Oceanology, Academia Sinica, 7 Nan-Hai pharmacology, and pollution control." (Dr. Road, Qingdao, Shandong, Peoples Republic M.L. Lizarraga-Partida, Centro de of China). Investigacion Cientifica y de Educacion Superior de Ensenada, Av. Espinoza No. "Our definition of marine biotechnology is 843, Apartado Postal 2732, Ensenada, Baja the use of biotechnology for studies of marine California, Mexico). organisms or the use of marine organisms for applications in the field of biotechnology." "Marine biotechnology is the application (Dr. Bert Ely, Director Institute for of marine organisms including their systems Biological Research and Technology, or processes for the manufacture of industrial University of South Carolina, S.C.). products and for the practical solution of problems created by human activity." "I would want to define marine (Milagrosa R. Martinez, Associate professor biotechnology as studies and development of and Director, Learning Resources Center, marine (aquatic) resources for human welfare University of Philippines at Los Banos, using the available biomolecular tools as well College, Laguna 3720, Philippines). as developing newer and better research tools for application and improvement- "Marine biotechnology is the science enhancement of our understanding of marine dealing with the study of marine organisms (aquatic) life in general." (Dr. S.O. (preferentially microorganisms and plants) at Emejuaiwe, Imo State University, P.M.B. a molecular level, specially on their genetic 2000, Okigwe, Imo State, Nigeria). structure and on the techniques that could be 104 WARINEBOTECHNOLOGYAND DEVELOPIN COUNTRIES used to modify or improve their genomes in "Tbe manipulation and/or use of all or order to produce substances (food, medicines, part of a specific marine biological system to etc) at a high quality and quantity level or to generate a desired product or products." degrade debris and undesirable substances in (Dr. Donald W. Renn, Senior Research by-products useful to mankind." (Dr. Fellow, FMC Corporation, Maine). Enrique C. Mateo, Fondo de Reactivacion del Sector Pesqueria, German Schereiber 198, "I shall define marine biotechnology as the Francia 726 - Miraflores, Lima, Peru). commercial exploitation of living marine organisms or their components. The 'Marine biotechnology as the application organisms will include microbes, and also of molecular biological techniques/methods to plants as well as animals; the later will the production or modification of potential encompass the application of molecular commercial products. This might include the biology and cell culture techniques." (P.M. use of marine species for the application, or Satheesh Seshaiya, Post Graduate Lecturer in the use of molecular bio-techniques in the Microbiology, Post Graduate Department of marine environment." (Dr. David L. Nebert, Microbiology, Sri Paramakalyani College, 29 Assistant Director for Research and West Car Street, Kallidaikurichi 627 416, Administration, Institute of Marine Science, Ta=ilnadu, India). University of Alaska-Fairbanks, Alaska). "'I would define marine biotechnology "The definition about marine simply as the application of the techniques of biotechnology managed by the Institute is the modern molecular biology to marine biology. same used in other Latin America countries It covers the use of these techniques to study and in Europe, any technology used to the biology of marine organisms as well as increase production where the final product exploit practical applications of molecules has commercial importance. In this sense, in derived from marine organisms." (Dr. USA and Canada this concept is much more Norman R. Wainwright, Director of restricted and its use has been applied to Research, Associates of Cape Cod, Inc., technology where only DNA is manipulated." Massachusetts). (Dr. Patricio Bernal Ponce, Executive Director, Instituto de Fomento Pesquero, Jose Domingo Canas 2277, Casilla 1287, Santiago, Chile). "Marine biotechnology is the manipulation of marine organisms to produce a beneficial product for humankind." (Dr. Kent S. Price, Associate Dean, College of Marine Studies, Lewes, Delaware). Abbreviatlons, acronyms and date note ADB Asian Development Bank AFDB African Development Bank AVHRR advanced very high resolution radiometer BLI (BL2, BL3 or BL4) biosafety level 1 (2, 3 or 4) of the NIH guidelines C Celsius CGIAR Consultative Group on Intemational Agricultural Raerch COMAR Coastal and Marine Program CZCS coastal zone color scanner DNA deoxyribonucleic acid DLR Deutsche Forschungs fur Luft und Raumfahrt EBRD European Bank for Reconstruction and Development E. coi Escherichia coU ECU European currency unit EIB European Investment Bank ERS type of European satellite EUREKA European Research Coordinating Agency FAO U.N. Food and Agricultural Organization FDA U.S. Food and Drug Administration GABA gamma-aminobutyric acid GEF Global Environmental Facility GESAMP Group of Experts on the Scientific Aspects of Marin Pollution GIAM Global Impacts of Applied Microbiology GIFT genetic improvement of farmed tilapias GLOSS global sea-level observing system GMAG Genetic Manipulation Advisory Group (United Xingdom) GOOS Global Ocean Observing System HCMM heat capacity mapping mission hGH human growth hormone IARCs International Agricultural Research Centers IBC Institutional Biosafety Committee ICES International Council for the Exploration of tha Sea ICGEB International Centre for Genetic Engineering and Biotechnology ICLARM International Centre for Living Aquatic Resourcos Management 1DB Inter-American Development Bank IFC International Finance Corporation IGLOSS Integrated Global Ocean System IHN infectious hematopoictic necrosis IMO International Maritime Organization IOC Intergovernmental Oceanographic Commission JERS type of Japanese satellite KFA type of Russian satellite LAL Limulus amebocyte lysate LANDSAT Type of U.S. satellite MAB Man and the Biosphere Programme MIRCEN Microbial Resource Center 106 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES MSS multispectral scanner NCI U.S. National Cancer Institute NGO nongovernmental organization NIH U.S. National Institutes of Health NOAA U.S. National Oceanic and Atmospheric Administration NRC U.S. National Research Council OECD Organization for Economic Cooperation and Developmnet OPEC Organization of Petroleum Exporting Countries PAN panchromatic PCR polymerase chain reaction PrepCom Preparatory Committee PSD private sector development RAC U.S. Recombinant DNA Advisory Committee rDNA recombinant DNA R&D research and development RFLP restriction fragment length polymorphism SAR synthetic aperture radar SLAR side-looking airborne radar SPOT type of French satellite toH trout growth hormone TM thermal mapper U.N. or UN United Nations UNCED U.N. Conference on Environment and Development UNDP U.N. Development Programme UNEP U.N. Environment Programme UNESCO U.N. Educational, Scientific and Cultural Organization UNIDO U.N. Industrial Development Organization U.S. or USA United States USDA U.S. Department of Agriculture W'HO World Health Organization XS multispectral (3-band) Data Note Dollars are U.S. dollars unless otherwise specified. Glossary of technical terms Aerobic requiring oxygen. Biotechnology a collection of processes and Amino acid any of a group of twenty chemicals techniques that involve the use of living that are linked together in various organisms, or substances from those organ- combinations to form peptides or proteins. isms, to make or modify products from raw Anabolism see metabolism. materials for agricultural, industrial or medical Anaerobic without oxygen. purposes. Antibody a specific protein molecule produced Bivalve one of a class of sessile or burrowing by an organism's immunological defense mollusks, including clams, mussels and system when it is challenged by a foreign oysters. substance (the antigen). The antibody Capability the ability to produce or apply a neutralizes the antigen by binding to it. particular set of scientific techniques or Antigen a substance that when introduced into an technologies. organism elicits from it an immunological Catabolism see metabolism. defensive response. Many living mi- Catalyst a substance that affects the rate of a croorganism or chemical agents can under chemical reaction but remains itself unaltered appropriate circumstances become antigens. in form or amount. Applied research experimental or theoretical Cell culture the propagation of cells removed work directed towards the application of from a plant or animal in culture. scientific knowledge for the development, Cell fusion combining nuclei and cytoplasm from production or utilization of some useful two or more different cells to form a single product or capability. hybrid cell. Bacteriophage (phage) a virus that attacks or Clone a group of genetically identical cells or colonizes a bacterium. Bacteriophages are organisms asexually descended from a specific; one type of phage will attack only common ancestor. In a cloned organism, all one species of bacteria. cells making up that organism have the same Basic research experimental or theoretical work genetic material and are exact copies of the that is undertaken to acquire knowledge of original. fundamental principles of phenomena and Cloning the use of genetic engineering to observable facts and that may not be directed produce multiple copies of a single gene or a towards a specific application. segment of DNA. Biodegradation the natural process whereby Crustacean one of the class Crustacea, which microorganisms break down organic breathe by gills and whose bodies are covered molecules. by shell or crust, including barnacles, crabs, Biodiversity the totality of the world's life lobster and shrimp. forms, ecosystems, and ecological processes, Culture the growth of cells or microorganisms in which can be characterized at the genetic, a controlled artificial environment. taxon (for instance, families and species), and Dispersant a substance that reduces surface ecosystem levels. tension of a floating pollutant, causing it to Bioremediation a technology that uses biological sink. activity to treat contaminated soil or water in Database a collection of data, defined for one or order to reduce or eliminate the more applications, which is physically located contaminant(s). and maintained within one or more electronic Biosafety in activities involving life forms or computers. their parts, the observance of precautions and Development the process of applying scientific preventive procedures that reduce the risk of and technical knowledge to the practical adverse effects. 108 MARINEBIOTECHNOLOGYAND DEVELOPNG COUNTRIES realization or enhancement of a specific fusion, plasmid transfer, transformation, product or capability. transfection and transduction. DNA deoxyribonucloic acid; the carrier of Halophilic tolernt of high concentrations of slt. genetic information found in all living Hazard the likelihood that an agent or substance organisms (except for a small group of RNA will cause immediate or short-term adverse viruses). Every inherited characteristic is effects or injury under ordinary circumstane coded somewhere in an organism's com- of use. plement of DNA. Host a cell whose metabolism is used for growth Emulsant a surface-active substance that allows and reproduction of a virus, plasmid, or other a normally immiscible liquid (for example oil) form of foreign DNA. to disperse or become mixed into a second Host-vector system compatible host/vector liquid (for example water). combinations that may be used for the stable Enzyme a special protein produced by cells that introduction of foreign DNA into host cells. catalyze the chemical processes of life. Hybridoma a special cell produced by joining a Eacherichia coli (E. coli) a species of bacteria tumor cell (myeloma) and an antibody that commonly inhabits the human lower producing cell (lymphocyte). Cultured hybri- intestine and the intestinal tract of most other doma produce large quantities a particular type vertebrates as well. Some strains are pathogen- of monoclonal antibodies. ic, causing urinary tract infections and Hydrocarbon one of a large and diverse group diarrheal dieas. Weakened strains are often of compounds, consisting of only carbon and used in laboratory experiments. hydrogen, constituting petroleum. Expression the translation of a gene's DNA Infection the invasion and sttling of a pathogen sequence by RNA into protein. within a host. Fermentation the anaerobic bioprocess in which Intellectual property the area of law yeasts, bacteria or molds are used to convert encompassing patents, trademarks, trade a raw material into products such as alcohols, secrets, copyrights, and plant variety acids or choeses. protection. Filterfeeder an organism that obtains its food by In vitro literally 'in glass"; pertaining to bio- straining water passing through some part of logical processes or reactions taking place in its body and recovering suspendod organisms. an artificial environment, usually the Filterfeeders include baloen whales, corals, laboratory. mussels and sponges. In vivo literally win the living'; pertaining to bio- Finfish true fish, as opposed to shellfish. logical processes or reactions taking place in Fraction a chemical agent or compound that may a living system such as a cell or tissue. be separated out by chemical or physical Metabolism the sum of the chemical and methods from a solvent containing a mix of physiological processes in a living organism in substances. which foodstuff are synthesized into complex Gene the fundamental unit of heredity. biochemicals (anabolism); complex Chemically a gene consists of ordered nucleo- biochemicals transformed into simple tides that code for a specific product or control chemicals (catabolism), and energy is made a specific function. available for the organism to function and Gene splicing the use of site specific enzymes procreate. that cleave and reform chemical bonds in Metabolite a substance vital to the metabolism of DNA to create modified DNA sequences. a certain organism, or a product of Genetic engineering a collection of techniques metabolism. used to alter the hereditary apparatus of a Microinjection the injection of DNA into a cell living coll enabling it to produce more or or cell nucleus using a fine needle under a different chemicals. These techniques include microscope. chemical synthesis of genes, the creation of re- Microorganism a microscopic living entity that combinant DNA or recombinant RNA, cell can be a virus, bacterium, or fungus. Glssawy of technicl tW 109 Mollusc invertebrate member of the phylum Risk management the process of determining Mollusca, including clams, mussels, octo- whether or how much to reduce risk through puses, sails and squids. regulatory action. Decisions usually depend on Monodonal antibody an antibody produced by data from risk aasssment and take into a hybridoma that recognizes only a specific account economic, ethical, legal, political and antigen. social factors. Nucleotide the fundamental molecule that makes RNA ribonucleic acid; found in three up DNA and RNA. Each nuclootide forms-messenger, transfor and ribosomal constituting DNA consists of one of four RNA. RNA assists in translating the genetic amino acids (adenine, guanine, cytosine or code of a DNA sequence into its comple- thymine) linked to the phosphate-sugar group mentary protein. deoxyribose; each nuclootide constitutingRNA Shellfish an indistinct term for marine inver- consists of one of four amino acids (adenine, tebrates, but commonly refers to any guanine, cytosine or uracil) linked to the crustacean or mollusc. phosphate-sugar group ribose. Synthesis the production of a compound by a Pathogen an organism that cauws diseas. living organism. Plankton microscopic organisms inhabiting sea Technology the scientific and technical water in high numbers. Plankton may be information, coupled with know-how, that is phytoplankton (microscopic plants) or zoo- used to design, produce and manufacture plankton (microscopic animas). products or gnerate data. P,asmnid small, circular, self-replicating forms of Toxicity the quality of being poisonous or the DNA often used in rDNA experiments as degree to which a substance is poisonous. aeoptors of foreign DNA. Trait a charactristic that is coded for in the Plasmid transfer the use of genetic or physical organism's DNA. manipulation to introduce a foreign plasmid Transduction the transfer of one or more genes into a host cell. from one bacteria to another by a Production the conversion of raw materials into bacteriophage (a virus that infects bacteria). products or components thereof through a Transfection the process in which a bacterium is series of manufacturing processos. modified in a way that allows the cell to take Real fime a characteristic of a system which up purified, intact viral or plasmid DNA. makes information available about a process so Transformation the introduction of new genetic quickly it allows the operator to act to change information into a cell using naked DNA (that the outcome of the process while it is still is, without using a vector). under way. Triploid having three haploid sets of Recombinant DNA (rDNA) tho hybrid DNA chromosomes in each nucleus. resulting from the joining pieces of DNA from Vector a trsnsmission agent, usually a plasmid or different sources. virus, used to introduce foreign DNA into a Risk the probability of injury, disease or death host cell. for persons or groups of persons undertaking Virs an infectious agent, containing either DNA certain activities or exposed to hazardous or RNA as its genetic material, which requires substances. Risk is sometimes expressed in a host cell for its replication. numeric terms (in fractions) or qualitative Wild-type an organism that is native to a locale terms (low, moderate or high). in nature. References Agellon, L.B., C.J. Emery, J.M. Jones, S.L. 1989. Applications of genetics to microalgae Davies, A.D. Dingle and T.T. Chen. 1988. production. In G.E. Pierce, ed., Developments Promotion of rapid growth of rainbow trout in Industrial Biotechnology, Volume 31, 271- Salmo gairdneri by a recombinant fish growth 274. New York: Elsevier. hormone. Canadian Journal of Fisheries and Bull, A.T., G. Holt and M.D. Lilly. 1982. Aquatic Sciences 45:146-151. Biotechnology: International Trends and Allen, S.K. Jr. 1988. Triploid oysters ensure Perspectives. Paris: Organization of year-round supply. Oceanus, 31(3):58-63. Economic Cooperation and Development. Alper, J. 1990. Oases in the oceanic desert. ASM Callegari, J-P. 1989. Feu vert pour les News 56(10):536-538. microalgues. Biofutur No. 76:25-40. Ansley, D. 1990. Cancer institute turns to cell Cardellina II, J.H. 1986. Marine natural products line screening. The Scientist 4:3. as leads to new pharmaceutical and Austin, B. 1989. Novel pharmaceutical agrochemical agents. Pure and Applied compounds from marine bacteria. Journal of Chemistry 58(3): 365-374. Applied Bacteriology 67:461-470. Carlton, J.T. 1989. Man's role in changing the Bailey, C. 1988. The social consequences of face of the ocean: Biological invasion and tropical shrimp mariculture development. implications for conservation of near-shore Ocean & Shoreline Management 11:31-44. environments. Conservation Biology Baker, R.N. 1991. Remote sensing of oil in the 33:265-273. marine environment: State-of-the art and Cell Tak and Matrigel. 1990. Trends in fiuture directions. European Science News Biotechnology 8(10):v. Information Bulletin 7:A1-A7. Center for Marine Science Research (CMSR). Baslow, M.H. 1977. Marine Pharmacology, 1990. Annual Report - July 1989 to July Huntington, N.Y.: Robert E. Krieger 1990, University of North Carolina at Publishing Company. Wilmington: Marine Biotechnology Program, Benemann, J.R. 1989. Microalgae products and Center for Marine Research. production: An overview. In G.E. Pierce, Chapman, D.J. and K.W. Gellenbeck. 1989. An ed., Developments in IndustrialBiotechnology, historical perspective of algal biotechnology. Volume 31, 247-256. New York: Elsevier. In: Algal and Cyanobacterial Biotechnology, Bialy, H. 1992. From the green to the green 1-27. New York: Wiley. revolution: Biotechnology and the Chen, T.T. and D.A. Powers. 1990. Transgenic international centers. Bioltechnology 9:900. fish. Trends in Biotechnology. 8(8):209-215. Bjurstrom, E.E. 1985. Biotechnology: Chowdhury, J. 1992. The rise of a dirty business. Fermentation and downstream processing. Chemical Engineering 98(11):44aa-44kk. Chemical Engineering 92:126-158. Cohen, S.N., A.C.Y. Chang, H.W. Boyer and Board on Science and Technology for R.B. Helling. 1973. Construction of International Development (BOSTID). 1990. biologically functional bacterial plasmids in Saline Agriculture: Salt-Tolerant Plants for vitro. Proceedings of the National Academy of Developing Countries. Washington D.C.: Sciences USA 70:3240-3244. National Academy Press. Colwell, R.R. 1986. Marine Biotechnology and 1982. Priorities in Biotechnology the Developing Countries, UNIDO report IS. Research for International Development: 593, January 7. Proceedings of a Workshop. Washington Cooksey, K.E. 1991. The fifth European D.C.: National Academy Press. Congress on Biotechnology: Marine Brown, L.M., Dunahay, T.G. & Jarvis, E.E. biotechnology. European Science News Rfefwnces 1 1 1 Information Bulletin No. 91-02:36-37. 4:34-40. Costa-Pierce, B.A. 1987. Aquaculture in ancient Fox, J.L. 1982. Complex structure of manne Hawaii. BioScience 37(5):320-331. toxin unraveled. Chemical & Engineering Costerton, W. and H.M. Lappin-Scott. 1989. News 60:19-20. Bdavior of bacteria in biofilms. ASM News Gain for fish farmers. 1991. New Yor* lnmes. 55:650-654. November 19:29. Couve, Andres. 1992. Interview with Chile's Gellenbeck, K.W. and D.J. Chapman. 1983. Fisheries Under Secretary Andr6s Couve on Seaweed uses: the outlook for mariculture. February 15 at 10:00 GMT. Santiago Radio Endeavour 71:31-37. Chikna Networ. Glenn, E.P., J.W. O'Leary, M.C. Watson, T.L. Crews, P. 1991. Personal communication, July Thompson and R.O. Kuehl. 1991. Salicornia 29. bigelovii Torr.: An oilseed halophyte for Cavas, I. 1989. Recent developments in coastal seawater irrigation. Science 251:1065-1067. aquaculture in the Asia-Pacific. INFOFISH Gold, B.D. and E.B. Shultz, Jr. 1986. Marine International No. 4:47-51. biofuel for nrual coastal and island Curtin, M.E. 1985. Trying to solve the biofouling communities in developing nations. In J. problem. Blo/technology 3:38. Twidell, I. Hounan and C. Lewis, eds., Cutler, A.J., M. Saleem and F. Georges. 1989. Energy for Rural and Island Communities IV, Can fish antifreeze proteins make frost 87-92. New York: Pergamon Pres. resistant plants? Plant Biotechnology Institute Gotfrit, C.W. 1990. Salmon farming in Chile: Bulletin, July: 1-3. from reality to dreams? INFOFISH DNAP plants, yeat expres flounder antifreeze. International No. 1:40-43. 1991. McGraw-Hill's Biotechnology Gourlie, B., Lin, Y., Price, J., DeVries, A.L., Newswatdc 11(9):2. Powers, D. and R.C. Huang. 1984. Winter Edwards, P. 1991. Integrted fish farming. flounder antifreeze proteins: A multigene INFOFISH International No. 5:45-51. family. Journal of Biological Chemistiy Engleking, H.M. and J.C. Leong. 1991. Subunit 259: 14960. vaccine trials using glycoprotein and Grassle, J.F. 1991. Deep-sea benthicbiodiversity. nuclooprotoin constructs of the rhabdovirus, BioScience 41(7):464-469. Infectious Hantopoietic Necrosis virus. In: Guerrero III, R.D. 1991. Farming tilapia in the 14th Annual AFS/FHS Meeting; 32nd Western Philippines. INFOFISH International No. Fish Disease Conference, 45. July 31 - 6:40-42. August 3, Newport, Oregon, OSU/Hatfield Hadwiger, L.A. 1988. Crustacean shells may aid Marine Science Center. diseased farm crops. Oceanus 31(3):73. Ferdouse, F. 1990. Shrimp from China. Harvey, S., I. Elashvili, J.J. Valdes, D. Kamely INFOFISH International No. 3:24-26. and A.M. Chakrabarty. 1990. Enhanced Fernandez-Pato, C. 1989. Mariculture removal of Exxon Valdez spilled oil from developments; environmental effects and Alaskan gravel by a microbial surfactant. planning. Ocean & Shoreline Management Bio/technology 8:228-230. 12:487-494. Heyward, A.J. and L.S. Hammond. 1990. First r-DNA 'clean-up bugs' to seek EPA Biotechnology for aquaculture. INFOFISH field-release OK. 1991. McGraw-Hill's International No. 6:48-51. Biotechnology Newswatch 11(11):1-3. Hightower, L.E. and J.L. Renfro. 1988. Recent Five-year red tide control campaign to begin (in applications of fish cell culture to biomedical Japanose). 1990. Kagaku Kogyo Nippo Tokyo. research. Journal of Experbnental Zoology Januay 22:11. 248:290-302. Food ad Agricultural Orgaization (FAO). 1991. Hoffman, J. 1990. Provesta forms marine biology Aquaculturm. Rome: FAO. company. Chemical Marketing Reporter . 1989. Review of developments in 238(22):27. aquacultmue. INFOFISH International No. Huglen, 6. 1991. Technologies for marine 112 MAAWE0TECNOLOGYAND DEVELOPNG COUNTRIES iiomt urvellance. Marine Industrial Development of exprssion vectos for Teduology Monitor No. 3:1-5. trmasgenic fish. Bioltechnology 8:1268-1272. Hum, H.H. 1991. Public hoalth aspects of Liana, E.G. 1991. Production and utilisaon of sefood conumption. INFOFISHInternational seaweeds in the Philippines. INFOFISH No. 3:27-32. International No. 1:12-17. lxilatvw jbr the Accekrated Transfer of Lovatelli, A. 1990. Bivalves: status, problems d BlemloVgy to the Ocean Sciencs. 1990. future in Asia. INFOFISH International No. Wahington D.C.: Joint Oceanogphic 2:20-24. ItitutioI bcorporated. Mabeau, S., 0. Vallat and D. Brault. 1990. De Intuational Cantr for Gonetic Engineering and l'Orient a I'Occident: Lea prmncipaux Bioechnology (ICOEB). 1990. AcviWty Reporn marches. Biofwur No. 88:24-29. - 1990. Tuate: ICOEB. Martello, A. 1991. Bioremediation: cleaning up Jackn, D.A., R.H. Symms and P. Berg. 1972. with biology and technology. The Sdcentst Biochemical method for insorting new genetic 5(1):18-19. infon into DNA of simian virus 40: Masters, B.A. 1990. Algae is a growth industy Circular SV40 DNA molecula containing for MD biotechnology company. Washington lambda phap gene and the galactose operon Post, December 31:5. of Escherichla coUl. Proceedings of the McConnell, D., S. Riazuddin, R. Wu and R.A. Natial Acadway of Scdmnca USA Zilinsku. 1986. Capability Building in 69:2904-2909. Biotechnology and Genetic Engineing iX Jap roudup. 1991. Bio/tedaology 9:130. Developing Countria, United Nations Joaupoit, H. 1991. Sponge - world production IndustrialDevelopmentOrpanizationdocuinInt and trade. INFOFISH International No. UNIDO/IS. 608. 2:21-27. McCoy, H.D., II. 1990. Worldwide aquaculture Kat, A. 1991. Opporlunitie. in India's marine production expected to double over next sector. INFOFISH International No. 1:24-29. decade. Genetic Engineering News 10:1. Kapwsindsi, A.R. 1990. Integrtion of trmnsgenic McElroy, S. 1990. The Japaneos pd mrkdet. fiak into aquaculturo. Food Reviews INFOFISH International No. 6:t7-23. International 63:373-388. McGuire, J. 1992. Personal communication, Klaur, A. 1986. U.S. to scroen marine February 28. organism for drugs. Bio/technology 4:684. Metting, B. 1989. Microalga applications in 1. 985. Food from the sea. agricultur. In G.E. Piere, d., Developments lIo/technology 3:27-32. in Industrial Biotechnology Volum 31, 265- K81ler, 0. and C. Milstein. 1975. Continuous 270. New York: Elsevier. culture of fused cells soereting antibody of Miller, H.I., R.H. Burris, R.H. and A.K predefined specificity. Nature 256:495-496. Vidaver. 1991. Regulation of biotechnology Komnberg, A. 1991. Biotech nightmare: Does (letter). Science 252:1599-1600. Cotus own PCR? Sdence 251:739. More on the oyster project. 1992. INFOFISH Kullmnberg, G. 1991. Personal communication, International. No. 1:33. Septmber 20. Morgan, D. 1990. Two firms race to derive Lemos from the golden snail. 1992. INFOFISH profits from mussels' glue. The Scientist, International. No. 1:33. 4(9):1. Linmonta, M. 1989. Biotechnology and the Third Morse, A.N.C. 1991. How do planktonic larvae World: Development srtegies in Cuba. In know where to settle? Amercan Scientist B.R. Bloom and A. Cerami, eds., Biomedical 79:154-167. Sdence and the Third World. Under the Morsw, D.E. and A.N.C. Morse. 1988. Chemical Volkano, 32S-334. New York: New York signals and molecular mechanisms: Larning Aademy of Scionces. from larvae. Oceanus 31(3):37-43. LUu, Z., B. Moav, A.J. Fams, K.S. Guise, A.R. National Research Council (NRC). 1990. Plant Kapuscini and P.B. Hackett. 1990. Biotechnology Research for Developing References 1 13 Countries. Washington D.C.: National Marine Biotechnology Conference, October Academy Press. 15, Baltimore, MD. . 1989. Field Testing Genetically Polysaccharides with high added value. 1991. Modified Organisms: Framework for French Technology Survey. December/ Decisions. Washington D.C.: National January:3. Academy Press. Portier, R.J. and S.1. Ahmed. 1988. A marine . 1985. Opportunities in Marine Science biotechnological approach for coastal and and Technology for Developing Countries. estuarine site remediation and pollution Washington D.C.: National Academy Press. control. Marine Technology Society Journal Newkirk, G. 1991. The world of oyster culture: 22(2):6-14. focus on Asia. INFOFISH International No. Powers, D.A. 1990. Marine and freshwater 6:43-46. biotechnology: A new frontier, (unpublished). 1989 FAO statistics. 1992. INFOFISH Interna- Practical course on genetically modified tional. No. 2:8. organisms. 1991. Genetic Engineering and Noga, E.J. 1987. Propagation in cell culture of Biotechnology Monitor. No. 34:2-3. the dinoflagellate Amyloodinium, an Primavera, J.H. 1991. Intensive prawn farming in ectoparasite of marine fishes. Science the Philippines: ecological, social, and 236:1302-1304. economic implications. Ambio 20(1):28-33. Novitsky, T.J. 1984. Discovery to Proctor, L.M. and J.A. Fuhrman. 1990. Viral commercialization: The blood of the mortality of marine bacteria and cyanobacter. Horseshoe crab. Oceanus 27:13-26. Nature 343:60-62. An ocean of viruses may affect global cycles. Ragotzkie, Robert A. 1988. The Sea Grant 1990. ASM News 56(12):632-633. concept - an introduction. Marine Technology Office of Technology Assessment (OTA). 1991. Society Journal 22(2):2-5. Bioremediation for Marine Oil Spills. Rechnitz, G.A. 1988. Biosensors. Chemical & Washington D.C.: U.S. Government Printing Engineering News 66:24-36. Office. Record year for shrimp farming. 1992. . 1984. Commercial Biotechnology: An INFOFISH International. No. 2:39. International Analysis. Washington D.C.: Renn, D.W. 1991. Personal communication, U.S. Government Printing Office. December 6. Olivera, B.M., J. Rivier, C. Clark, C.A. Ramilo, . 1990. Seaweeds and biotechnology - G.P. Corpuz, F.C. Abogadie, E.E. Mena, inseparable companions. Hydrobiologia S.R. Woodward, D.R. Hillyard and L.J. 204/205:7-13. Cruz. 1990. Diversity of Conus neuropeptides. . 1986a. Marine algae biotechnology: Science 249:257-263. Possibilities and realities. In Board on Science Olson, B.H., R. McCleary and J. Meeker. 1991. and Technology for International Background and models for bacterial biofilms Development, ed., Workshop on Marine Algae formation and function in water distribution Biotechnology: Summary Record, 53-67. systems. In C.J. Hurst, ed., Modeling the Washington D.C.: National Academy Press. Environmental Fate of Microorganisms, 255- . 1986b. Uses of marine algae in 285. Washington D.C.: American Society for biotechnology and industry. In Board on Microbiology. Science and Technology for International Peacock, F. 1991. Personal communication, April Development, ed., Workshop on Marine Algae 15. Biotechnology: Summary Report, 16-29. Pearls from freshwater mussels. 1992. INFOFISH Washington D.C.: National Academy Press. International. No. 1:35. Richards-Rajadurai, N. 1990. Carrageenan- Polne-Fuller, M., A. Rogerson and A. Gibor. multipurpose gum from the sea. INFOFISH 1991. Trichosphaerium 1-7, a marine amoeba International No. 5: 18-22. which digests polyethylene and 'saran wrap". Robinson, P.K. 1985. Phycotechnology. Paper presented at the Second International Industrial Biotechnology 42(5):73-78. 114 MARINE BIOTECHNOLOGY AND DEVELOPING COUNTRIES Rodrigue, D.C., R.A. Etzel, E. Porras, O.H. Technical Q&A. 1991a. INFOFISH International Velasquez, R.V. Tauxe, E.M. Kilbourne and No. 6:59-60. P.A. Blake. 1990. Lethal paralytic shellfish Technical Q&A. 1991b. INFOFISH International poisoning in Guatemala. American Journal of No. 5:69-70. TropicalMedicineandHygiene42(3):267-271. Thermostable DNA polymerase. 1990. Trends in Rojas, H. 1991. Waste contaminates marine Biotechnology 8(10):v. products (in Spanish). El Mercurio Santiago, Thorne-Miller, B. and J.G. Catena. 1991. The Al. May 2. Living Ocean: Understanding and Protecting Ruying, S. and W. Qinguin. 1992. Laminaria Marine Biodiversity, Washington D.C.: Island culture in China. INFOFISH International No. Press. 1:40-43. Transgenic fish experiment gets green light. 1990. Sasson, A. 1988. Biotechnologies and Biotechnology Notes 3(10):1-2. Development, Paris: UNESCO. Trigo, E.J. and W. Jaffe. 1990. Biosafety Scheuer, P.J. 1990. Some marine ecological regulations in the developing countries. phenomena: Chemical basis and biomedical Genetic Engineering and Biotechnology potential. Science 248:173-177. Monitor No. 30:46-52. Seaweed for the desert. 1991. UNISTAR Briefs, 2. Tucker, J.B. 1985. Biotechnology goes to sea. Selwood, T. 1992. Barramundi skin. INFOFISH High Technology 5:34-44. International No. 1:26-28. United Nations Conference on Environment and Shang, Y.C. 1991. Sponge farming in Development (UNCED). 1991. Biotechnology: Micronesia: Some economic aspects. Options for Agenda 21. Report of the INFOFISH International No. 2:42-44. Secretary-General of the Conference, Shariff, M. and R. Subasinghe. 1990. Health Document #A/CONF. 151/PC/ 42/Add.5, July aspects of Asian aquaculture. INFOFISH 9. International No. 5:35-38. . 1992. Environmentally sound Sindermann, C.J. 1986. Strategies for reducing management of biotechnology. In Agenda 21 risks from introduction of aquatic organisms: (Chapter 16). A marine perspective. Fisheries 11(2):10-15. United Nations Development Programme Singh, T. 1991. Malaysia enjoys the seabass (UNDP). 1989. Plant Biotechnology including boom. INFOFISH International No. 2:45-48. Tissue Culture and Cell Culture, New York: Singleton, F.L. and J.G. Kramer. 1991. UNDP. Biotechnology of marine algae: Opportunities United Nations Environment Programme (UNEP). for developing countries. Genetic Engineering 1991. UNEP-Sponsored Programmne for the and Biotechnology Monitor No. 25:83-90. Protection of Oceans and Coastal Areas, Smith, A.C. 1988. Marine animals: Clear models UNEP Regional Seas Reports and Studies No. for medical science. 7he Scientist 2:18-19. 135. Stone, W.H. 1987. Defining biotechnology . 1990. GESAMP: 7he State of the (letter). Bioltechnology 5:1339. Marine Environment, UNEP Regional Seas Suttle, C.A., A.M. Chan and M.T. Cottrell. Reports and Studies No. 115. 1990. Infection of phytoplankton by viruses United Nations Industrial Development and reduction of primary productivity. Nature Organization (UNIDO). 1991. Voluntary Code 347:467-469. of Conduct for the Release of Organisms Into Swaminathan, M.S., ed. 1991. Biotechnology: the Environment, Vienna: UNIDO. Reaching the Unreached, Madras, India: . 1981. Exchange of Views with Experts Center for Research on Sustainable on the Implications of the Advances in Genetic Agricultural and Rural Development. Engineering for Developing Countries, Taylhardat, A.R. 1989a. The ICGEB: A center UNIDO document IS. 259. of excellence Part I. Biofutur No. 77:45-49. Updegraff, D.M. 1991. Background and practical Taylhardat, A.R. 1989b. The ICGEB: A center applications of microbial ecology. In C.J. of excellence Part II. Biofutur No. 79:53-59. Hurst, ed., Modeling the Environmental Fate References 1 15 of Microorganisms, 1-20. Washington D.C.: countries. In Capability Building in American Society for Microbiology. Biotechnology and Genetic Engineering in Vining, L.C. 1991. Functions of secondary Developing Countries, 61-75. United Nations metabolites. Annual Review of Microbiology Industrial Development Organization document 44:395-427. # UNIDO/IS.608. Weiner, J. 1985. Marine biotech in the Negev Yaoqing, J. and C. Xiongfeng. 1990. Study on desert. Bioltechnology 3:39. antifreeze protein in fishes. II. The cloning of Weiss Jr., C. 1985. The World Bank's support antifreeze protein gene cDNA of for science and technology. Science Pseudopleuro-nectes yokohamae and its 227:261-265. expression in E. coli (in Chinese). Yichuan Welcomnme, R.L. 1986. International measures Xuebao [ACTA Genetica Sinical 17(3):202- for the control of introductions of aquatic 210. organisms. Fisheries 11(2):4-9. Yap, W.G. 1990. Backyard hatcheries take off in World Bank. 1992. The World Bank Annual Jepara. INFOFISH International, No. 2:42-47. Report 1991. Washington, D.C. Zilinskas, R.A. 1993. Bridging the gap between 1991a. Agricultural Biotechnology: research and applications in the Third World. 7he Nexi 'Green Revolution'?. Technical World Journal of Microbiology and Paper No. 133. Washington, D.C. Biotechnology 9(2):145-152. . 1991b. Fisheries and Aquaculture . 1989. Biotechnology and the Third Research Capabilities and Needs in Asia. World: The missing link between research Technical Paper Number 147. Washington, and applications. Genome 31(2):1046-1054. D.C. Wu, R. 1986. Building biotechnology research and development capability in developing Distributors of World Bank Publications ARGENT1NA The Middle East Obser KENYA SOUTH AFRICA, BOTSWANA Carios Hirsch, SRL 41, Sherif Stet Africa Book Semrice (EA. Ltd. For sitgt$ title Calri. Gunmes Cairo Quaran House, Mfangano Street Oxford Univerity Pi Florida 165 4th Floor-Ofc 453/465 P.O. Box 45245 Southern Afri 1333 Buenos Air FINLAND Nairobi P.O. Box 1141 Akateerninen Kiryskauppa Cape Town 8000 AUSTRALIA, PAPUA NEW GUINEA. P.O Box 128 KOREA. REPUULICOF FIJI lSOLOMON ISLANDS, SF-4010 HeLainki 10 Pan Korm Book Corporatuion For eucniampio oiievs VANUATU, AND WESTERN SAMOA P.O. Box 101, Kwangwhamun International Subscription Service DA. Books & Iourn als FRANCE Seoul P.O. Box 41095 648 Whitehorse Road World Bank Publications Craigiall Mitcham 3132 66. avenred'ltna MALAYSIA Johanneeburg2024 Victoria 7Sl16Paris UnivesityofMalaysCooperative Bookahop, Limited SPAIN AUSTRIA GERMANY PO. Box 1127, JaLan Pantel Baru Mundi-Prena Libro1 . SA. Cerold and Co. UNO-Verlag 59700 Kuala Lunpur Caselo 37 Crben 31 Poppeladorfer AlleeSS 28D0i Madrid A-1011 Wien D-5300 Bonn I MEXiCO iNlVTEC Librrti Inteacdoai AEDOS BANGLADESH HONG KONG, MACAO Apartedo Post]l22-SW Consrel de Cent 391 Micro Indus Development Aaia 2000 Ltd. 14060 Tialpan. Mexico D- . O0 Barcelona Asistance Sodety (Ml DAS) 46-48 Wyndham Steet HOsUeS, Road 16 Winning Centre NETHERLANDS SRI LANKA AND THE MALDIVIS Dhanrmondi R/Area 2nd Flior De LindtboomAlnOr-Publikatis Lake House Bookshop Dhaka 1209 Central Hong Kong P.O. Box 202 P.O Box 244 7480 AE Haksbene 100. Sir Chithrmpalam A. Breech q7tie: INDIA Gardiner Mawatha 156, Nur Ahmed Sark Allied Publishers Privat Ltd. NEWZEALAND Colonibo 2 Chilatgong 400 751 Mount Road EBSCO NZ Lid. Mad ro - 600 002 Private Mail Bag 99914 SWEDEN 76 KD.A Avenue New Market For Nio* htl Kolna 9100 Bre, ffice: Auckland FritznFPackbokstoretet 15 IJN. Heredia Marg Regennpgatsn 12, Box 163r6 BELCIUM Ballard Estate NIGERIA S-103275lcckholm jean D Lannoy Bombay - 400 038 University Pi Limited Ay. du Roi 202 Three Cowns Building Jericho For sndsch r snixn 1060 Bruas 13/14 Asf Ali Road Private Mail Bag gSiL Weruorgren-WilliamaAB New Deihi -110002 fbadan P. Q Box 1305 CANADA 5-171 25 Solnn La Diffusrur 17 C)ittaranan Avenue NORWAY CP. 85S1501.Brue Ampere Calcutta -70072 Noreni normatron Cents SWIERLAND Boucherville, Qu6bec Book Department For sigle title: 14B SE6 Jayadeva Hotel Building P.O. Box 612S Etterstad Libraine Payot 5th Main Road. Gandhinaur N-0602 Oslo 6 1, me de BourS CHILE Bngaore - 560 009 0H41002 LausRnn Invertec IGT S.A. PAKISTAN Arnenco Vespudco Norh 1165 3-5-1129 Kachiguda Mizne Book Agency For racrscrtiv, orders: San,iago Crow Road 65, Shahrh_i-eQuaid--Azamn Libramr Payot H yderabad -500027 Pa Box No. 729 Srvie ds Abonnomente CHINA Lahor 54000 Cas postalo 3312 China Finarcial & Economic Prarthana Flate, 2nd Floor C1 1002 Lausane Publishing Hous Near Thakore Beu& Navrngppura PERU 8. Da F. Si Dong Jie Ahmnedabad - 380 009 Editorial Desrrolo SA TANZANIA Beijing Aparfdo 3824 Oxford Universty Pres Palala House Lime I P.O. Box 5299 COLOMBIA 16-A Ashok Murg Makteba Road Infoenlare Ltde. Lucknow -226C01 PHIUPPINES De Salaarn Apartdo Aeroo 34270 International Book Cener Bogota D. . Central Bazaar Road Suite 1703, CGtyland 10 THAILAND 60 Bajaj Naa Condomrouum Tower I Central Departnent Store COTF D'TVOIRE Nagpur 440 010 Ayaia Avenue, Corns H.V. doeL 306 Silom Road Centre d'Edition et do Diffusion Cota Extnsum Bangkok Africaines (CEDA) INDONESIA MAktb, Metro Manila 04 B.P. 541 Pt Indira Limited TRINIDAD & TOiBAGO. ANTIGUA Abidjan 04 Plateau Jalan Borobudur 20 POLAND BARBUDA, BARBADOS, P.O. Box 181 Inntertional PublilshingSrice DOMINICA, GRENADA, GUYANA, CYPRUS Jakarta 10320 Ul. PteknA31/37 JAMAICA, MONTSERRAT, ST. Cents of Applied Resarch 00-677 Warzawa KITTS & NEVMS, ST. LUCIA, CyprusColleg* IRELAND ST. VINCENT & GRENADINES 6, Dogenes Street Engotri Covernment SuppUes Agery For osc s otder-: Systmetics Studis Unit P.0- Box 20D6 4-5 Harrcurt Rood IFSJournals 89 Watts Strdee Nicoxia Dublin 2 LI. Okrera 3 Curepe 02-916 Warsawa Trinidad, Wet Indil DENMARK ISRAEL Sa:mfundsLitteratur Yozmot 1iterature Ltd. PORTUGAL TURKEY R-enoemns AIM II P.O. Boa 56055 Livruria Portuogi Infotol DK-1970 FrederiksborgC Tel Aviv 61560 Rua Do Carm. 70-74 NarababFe Sok. No.1S 1200 Usbon Cagalogiu DOMINICAN REPUBIUC ITALY lsatnbul Editorm Taller, C por A. Liros Commniionarie Sxnroni SPA SAUDI ARABI A, QATAR Restauractin e lsabel la Catelk 309 Via Duc Di Calbrie, 1/I Jarir Book Store UNITED KINGDOM ApertadodeCorreos2l90Z-l CasllaPostale552 P.O. 8o-7196 MicroinfoLtd. Sano Domingo 50125 Firnze Rlyadh 11471 P.Q Box3 Altort. Hampshire C1I34 2PC EGYPT, ARAB REPUBUC OF JAPAN SINGAPORE TAIWAN, EnAond Al Ahmm Easrn Book Servic MYAMAIkBRUNEI Al C-aIa Stee Hono 3-Chorn-, Bunkyo-ku 113 Inforrotiron Publications VENEZUELA Cairo Tokyo Pr. nbt, Lid. Libreria del EtA Goldn Wheel Building Aptdo. 60.337 41, KalJlng Puddins a04-03 Caracas 1060-A Sing.pore 1334 Recent World Bank Discussion Papers (contitnuied) No. 180 Clhina's Reforn Experience to Date. Peter Harrold No. 181 Coinbatritng AIDS and Orier Sexually Transmnitted Diseases in Africa: A Review oft/ic lVorld Bank's Agendafor A-trio Jean-Louis Lamboray and A. Edward Elniendorf No. 182 Privatization Problems at Industry Level: Road Haulage in CentIral Europe. Esra Bennathan and Louis S. Thompson No. 183 Participatory Development and the World Bank: Poteninal Directionsfor Chlange. Bhuvan Bhatnagar and Aubrey C. Williams, editors No. 184 AgricuIltral Research in Southern Afica: A Framneiorkfor Action. Andrew Spurling, Tcck Y. Pce, Godxin Mkamanga, and Christopher Nkwaanyania No. 185 Military Expenditure and Economic Developmetit: A Symposium on Researc/i Issues. Edited by Gcoffrey Lamb witlh Valeriana Kallab No. 186 Efficiency and Suibstituition in Pollution Abatetnetnt: Tliree Case Stu4dies Dennis Andcrson and William Caveiidish No. 187 The State Holding Companty: Issues and Options. Anjali Kumar No. 188 Indigenous lViews of l and and the Environment. Shclton H. Davis, editor No. 189 Poverty, Population, and t/ie Envirotnment. Stephcn D. Mink No. 190 Natural Gas im Developingq Connitries: Eialuaating the Betiefits to the Environment. John Hoiier No. 191 AppropriateMacroeconomic Managenierit in Indonesia's Open Economy. Sadiq Ahmcd No. 192 Telecomnmunnications: World Batik Experic.nce and( Strategy. Bjorm Wellenius and othcrs No. 193 Infornation Systems Strategies for Public Financial Mkfana getntit. H-wel N1. Davics, Ali Hashiiii, and Eduardo Talero No. 194 Social Gainsfromn Femnale Ediucation. A Cross-Nationial Study. K. Subbarao and Laura Rancv No. 195 Towards a Sustainable Developmtnent: The Rio dejaneiro Study. Edited by Alcira Kreimcr. Thereza Lobo, Braz Menezes, Mohan Munasinghe, and Ronald Ilarkcr No. 196 Eastern Europe in Transitionj: Fromt Recession to Crowth?: Proceedings of a Conjfereuicc on the Macroeconomic Aspects o/ Adjiustmnet,t Cosponsored by t/ie Intertiation al Mlonetary Fnid and the WVorld Batik. Edited by Mario I. Blcjcr. Guillermo A. Calvo, Fabrizio Coricelli, and Alan H. Gelb No. 197 Korean Industrial Policy: legacies of the Past an d DirectionsJ/or the F ture. Danny M. Lcipziger and Petcr A. Petri No. 198 Exporting Hig/i- Value Food Comttiodines: Siaess Storiesfrotm Developing Conmitrics. Steveni Ni. Jaflcc withi thc assistancc of Peter Gordon No. 199 Boreower Owtners/ip of Adjiustnet Programis arid the Politic-al Economniy of Reforn. John H. Johnson and Sulainian S. Wasty No. 200 Social It!frastnicture Constnrctiotn ii thie Sahiel: Optionsfor lmnprovinm Currenit Practices Bemard AbeillU and Jean-Marie La-ntran No. 201 Urbanizationj, Agricultural Development, and Land Allocationi. Dipasis Bhadra and Antonio Salazar P. Brandao No. 202 Koreati Induistrial Policy: Leacies of the Past and Directionsfor the Future. Danny M. Leipziger and Peter A. Petri No. 203 Pov'erty Reductioti in East Asia: The Silemit Revolution. Frida Johansenl No. 204 Maanagitig tlhe Civil Senrice: Tlhe Lessotis of Reforn ini Industrial Countries Barbara Nunberg No. 205 Designing a Systern of Labor Market Statistics amid Iifortnation. Robert S. Goldfarb and Arvil V. Adaiis No. 206 Infortnatiot Technology in World Bank Lending: Increasing t/ie Developmnetit lmnpact. Nagy Hanna and Sandor Boyson No. 207 Proceedings oJfa Conteretnce oj Cirreticy Substitutiion and Currency Boards. Nissan Liviatan No. 208 Developin.g Effective Employmnetnt Services. David Fretvell and Susan Goldberg No. 209 Evolving Legal Franmeworksfor Private Sector Development iti Central amid Eastern Euirope. Cheryl W. Gray The World Bank Headquarters European Office Tokyo Office H 1818 H Strect, N.W. 66, avenue d'1ena Kokusai Building Washinigton, D.C. 20433, U.S.A. 75116 Paris, France 1 -1 Marunouchi 3-chome Chiyoda-ku, Tokyo 1 00, Japan Telephonie: (202) 477-1234 Telephone: (1) 40.69.30.00 Facsimile: (202) 477-6391 Facsimile: (1) 40.69.30.66 Telephone: (3) 3214-5001 Telex: wui 64145 WORLDBANK Telex: 640651 Facsimile: (3) 3214-3657 RCA 248423 WORLDBK Telex: 26838 Cable Address: INTBAFRAD WASHINGTONDC 12590 ENV 100 0-8213-2590-6 MARINE BIOTECHNOLOGY & D J11 111111111 111111 400000009285 $9. 95 ISBN 0-8213-2590-6