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210 E2lWorldK Bank Discussion Papers

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Raymond A. Zilinskas Carl GustafLundin Public Disclosure Authorized Recent World Bank Discussion Papers

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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 comlmuleintwithinl 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 on1the 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, RkaymonidA. 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-buildingin 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, profoundlyaffecting 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'sproductiveactivities-whetherthey commercial products or processes. This be agriculture,health delivery, environmental cooperationand support is crucial in biotech- remediation, industry or energy production. nology both (I) because of the inherentrela- 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 introducesa 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 continuedscienti- both mankind and the environment. fic involvementin the industrialdevelopment "The BiologicalAge" can alsobe expected of that process (a situationquite 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 automobileindustry 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-tropicaland tropical operation, as complex as it is, can be done climatesthat 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 biotechnologyindustry involves the ma- (2) most developingcountries have sufficient nipulation of living and therefore dynamic grounding in the biological sciences and biological systems to achieve products or technologiesthat underliebiotechnology to be processes, trained scientiststhat know these able to build capacity even in advancedtech- 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, developmentand 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 considerablecommit- 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 supportingspecial training for scientistsor investigate and solve local environmental industrial managers in technology transfer, problems, and to collaboratein solvingglobal biosafety and intellectualproperty issues. problems as well. To date, World Bank involvementin bio- Fundamental to the development of bio- technology is rather limited, and entering an technology,and to any cooperativeventures, emergingfield at its beginningis a departure however, is that governments, scientists, from previous Bank practice. There are privatesectors, and internationaldevelopment several reasons for proposing this initiative. organizationsmust first learn of the potential First, the marine area is relatively neglected and problems of this new scientific endeavor, in Bank investments-only a smallfraction of especially as they relate to the home coun- that channeledto land-basedefforts. 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 biotechnologyfor economicdevelopment 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 developmentorganizations. The Environment could be one of the means toward sustain- Department presents this paper to further able, environmentallysound, developmentof these purposes and to encouragediscussion of ocean resources. The remediationof polluted how the Bank can best lend its support to this coastalwaters and beachesalso could be done emerging area of science and technology, to via marine biotechnology. Third, solving the benefit of sustainabledevelopment. global enviromnentalproblems will require participationfrom all countries, includingthe developing world. Without worldwide co- operation,such problemsas the 'greenhouse" 7t effect, pollution of the high seas, destruction of coral reefs, loss of marine biodiversityin MohamedT. El-Ashry general, and certain water-borne infectious Chief EnvironmentalAdviser diseases will defy solution. In the process of to the President and buildingtheir capabilityin marinebiotechnol- Director of the EnvironmentDepartment Contents

Acknowledgements vii

Introductionby 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 environmentaldamage and destruction 2 Detecting and monitoringoceanic events 3

Chapter 2 A primer in biotechnology 6 Classical biotechnology 6 Advancedbiotechnology 7 Biotechnologyand developingcountries 9 Safety and biotechnology 10 Biosafetyand marine biotechnology 15

Chapter 3 Marine biotechnologyand its sub-areas 24 Definition of marine biotechnology 24 Marine biotechnologyresearch and applications 24

Chapter 4 Options in marine biotechnologyfor developingcountries 55 Existing scientific and technicalcapabilities 55 Applicationsof marine biotechnology in the short and medium terms 57

Chapter 5 Buildingcapability in marine biotechnology 66 Building R&D capability 66 Applying research 69 Industry and biotechnology 69 The role of governmentin building capabilityin biotechnology 70 Discussion 71

Chapter 6 The experience of major internationalagencies in marine biotechnologyand related areas 73 Food and Agricultural Organization 73 Inter-AmericanDevelopment Bank and other DevelopmentBanks 74 IntergovernmentalOceanographic Commission 74 vd CONTENTS

InternationalCentre for GeneticEngineering and Biotechnology 75 InternationalMaritime Organization 76 United Nations Conferenceon the Environmentand Development 76 United NationsDevelopment Programme 77 United NationsEducational, Scientificand Cultural Organization 77 United NationsEnvironment Programme 78 United Nations Industrial DevelopmentOrganization 78 World Bank 80 Discussion 83

Chapter 7 ExploringWorld Bank options for investments in marine biotechnology 84 Science and technologylending 84 Support of environmentalobjectives 86 Support for private sector development 87

Chapter 8 Conclusion 90

AppendixA Marine biotechnologyand related R&D institutions in developingcountries 91

AppendixB Special equipmentrequirements for advancedbiotechnology 100

AppendixC Definitionsof marine biotechnologyby scientists in industrialand developingcountries 102

Abbreviations,acronyms and data note 105

Glossary of technicalterms 107

References 110 Acknowledgements

This report would not have been possible donesia), Dr. MilagrosaR. Martinez(Philip- without the assistance, cooperationand 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 organizationsthey 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 technicalprogress 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 mentalorganizations 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 biotechnologyor related areas activitiessponsored 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. MontenegroArcila (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/technicalreviewers 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 especiallydirected Esther FernandezA. (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- MoazzamKhan (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 applicationsof marine proven to be of potential pharmaceutical biotechnologyand is focused on developing value. However, I estimate that far less than countries of the world. It makes clear that 1 percent of potentiallyuseful chemicalsfrom biotechnology'spotential for remediatingeco- marine living sources have been screened so nomic and social problems of developing far. The chemical compoundsthat remain to countries is extraordinary. However, it is be discoveredrepresent one of earth's great- important to realistically assess what can be est treasures. done in selected areas of biotechnology. In additionto marinepharmaceuticals, en- Biotechnologyis not a panaceato cure all ills hancementof fisheries via aquaculture offers of mankind, especiallyproblems arising from great value. Overharvesting of the world populationincrease, overutilization of natural oceans is recognized as a global threat. The resources, and underutilization of human ability to produce transgenicfish 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 industryand offers some clear recommen- earth's surface. Developingcountries are pre- dations for enhancementof marine fisheries. dominantly riparian and, for the most part, Other areas of marine biotechnologythat 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 incomein additionto 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 extraordinarybiological 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 animalsin the marineenvironment, as well as these techniques can greatly benefit those seaweedsand marine plants, have been inten- countries where environmentalcontamination sively studied. Results from this research has reached threatening proportions.It is not demonstrate complex chemical interactions yet clearhow bioremediationcan be deployed betweenthese bacteria and their hosts includ- in operationsto treat offshore or open ocean ing systemsof elaborate signallingand terri- spills. torial marking. In studying the chemicals The potential of marine biotechnologyin 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 biotechnologybe applied in the technology is that the genetic capability of marineenvironment, 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 manipulatedand magnified aquaculture. in the laboratory, without depleting the re- The informationprovided in this document source in the natural environment. lays the groundworkfor an effective and effi- It has been stated that the oceans are cient design of a plan of action for devel- underutilizedin their capacity for feeding the oping marine biotechnologyin 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 conditionsfor their fish farming and seaweed farming) are limit- populationsare likely to be the main benefi- ed to maximumyield much as is land agricul- ciaries. ture. However, with the applicationof genet- ic engineering,the productivityof the coastal areas can be greatly improved. The forces at play are the increasing populationsof 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 BiotechnologyInstitute Note from the authors

This report focusseson a rapidly emerging niques can be used to break down pollutants, science-basedtechnology-marinebiotechnol- to alleviate environmentaldamage. ogy. The term 'marine biotechnology"differs The intent of this report is to introduce in meaning among scientists. We think of marine biotechnologyto nonspecialists,clari- marine biotechnology as a collection of re- fy its relevance to developingcountries, and search and developmental activities in the outline the role of the World Bank and of biological, chemical and environmentalsci- other internationalagencies 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 consideredin isolation; after all, it facilities, located mostly in the United States is that intersect where biotechnology meets and Japan are dedicatedto 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-relatedareas. they support and an introductionto the wider "Emerging," for the purpose of this re- field of biotechnologyare useful as a starting port, means that the technologyis at a stage point. Thus, Chapter 1 sets the stage for the in its developmental cycle when practical marine part of marine biotechnology by applicationsengendered by the technologyare briefly describing and discussing certain being identifiedand laboratoryprocesses 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 technologyis one that the of a review of its history, a presentation of public and its representativesbegin to recog- recent developments in this field, and an nize as having the potential to generate new explanation of its "classic" and "advanced" scientific knowledgeand produce useful new components. Special attention is given to products and processes. Marine biotechnolo- advancedbiotechnology, includingits work- gy research has produced a few applications force and equipment requirements and the to date, but its potential economiceffects are possiblerisks biotechnologyresearch, 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 possessingterrito- review; it defines marine biotechnologyand ries of subtropicaland tropical marine waters provides a description of major sub-areas that shelter a large diversity of estuarineand (suggestions for how marine biotechnology marine life. At the same time, large areas of should be defined are found in AppendixC). their marine and coastal environmentssuffer Our consideration of marine biotechnology from the detrimental effects of manmade continuesin Chapter 4, where optionsfor de- pollutants. The largest promises of marine veloping countries in this field delineatedand biotechnology are thus in two areas. First, analyzed. Specifically,each of the nine sub- some of its techniquesmay be deployed for areas of marine biotechnologyis assessed in the sustainableexploitation of naturalresourc- terms of promise for developingcountries in es under environmentallysound conditions; the short and mediumterm and the degree of and second, other marine biotechnologytech- difficulty inherent in capabilitybuilding (we xl NO TE FROM THEA UTHORS

list institutes in developing countries whose solving and for economic development. We work programs encompassmarine biotechnol- conclude the substantive part of the report ogy or related areas in Appendix A). Meth- with a short essay on why it is importantnow ods for building capability in biotechnology for developing countries to commit to capa- and in marine biotechnologyare explainedin bility building in marine biotechnologyand Chapter 5 (somespecific requirements for ad- why intergovernmentalorganizations should vanced biotechnology R&D are listed in assist in this endeavor. AppendixB). Examplesof projects in marine This report is intended mainly for World biotechnologyand related areas supportedby Bank professionals who in the future may major intergovernmentalagencies, 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 biotechnologycapability building in develop- of technical terms is provided. Those who ing countries, includingensuring that results wish to delve more deeply into the subject from research are applied in nationalproblem matter may consult the references. ExecutiveSummary

The tremendous diversity of life in tropi- Marine naturalproducts 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 organismssecrete compoundsthat help them Haiti, Indonesia and the Philippines, as well survive and that incidentallyhave 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-healingchemi- Marine biotechnology,which is definedas cals will certainly be found. This field has "the applicationof scientific and engineering substantial possibilities for growth since principles to the processing of materials by fewer than 1 percent of marine species have marine biologicalagents 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 pollutantsand wastes in soil or water to cell culture, biosensorsand terrestrial agricul- harmless or less toxic end products. The ture. microbesused in bioremediationhave usually been recovered from natural sites but have Aquaculture had their naturalcapability for breaking down pollutants enhanced through research and Marine biotechnologymay benefit aqua- development (R&D). Most bioremediating culture in two major ways. First, its research microorganisms do not survive after the techniquescan enhancea culturedorganism's substance they feed on has been destroyed. growth rate, procreation proficiency, disease Because under normal circumstancesbiore- resistance and ability to endure adverse envi- mediation causes less damageto 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 conventionaltechniques for clean- Second, throughbiotechnology vaccines may ing polluted harbors and waterways, as well be developed against bacterial and viral as decontaminatingestuaries, mangrovesand diseases that commonlyafflict 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 BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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 antibodywill, for instance,bind itself to only speedand 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 monoclonalantibod- settling. However, toxic paints pose health ies or DNA probes are being used to quickly hazards to workers and pollute seawater. detect and accuratelyidentify 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 encrustationby marine organisms. identifying and monitoring substances and pathogensin the oceans, enablingscientists 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 Terrestrialagriculture subdivide much like bacteria. Cultured cells can be used to generate whole plants or they Marine biotechnologyresearchers seek to can be directed to synthesize natural prod- transfer characteristics inherent to marine ucts. Contrary to the chemical synthesis of animalsand plants to their terrestrial counter- compounds,cell cultureproduction is energy- parts. For example, the winter flounder conserving and essentially nonpolluting. survivessub-zero temperatures that would kill Many terrestrial plant cell culture systemsare 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 altitudesor northern latitudes. Anoth- valuable marine natural products. er example involves the world's most salt- tolerant plant, a microalgalspecies 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 chemoreceptoror 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 complexityfrom simple salts to pheromones. enormous promise for developing countries, In the second kind of sensor, an immunologi- that promise cannot be transformedinto real- cal molecule, the recognition element is a ity until capability is built in all of the sup- ExecutiveSummary 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 infrastructureto undertake wide- sharing of intellectualproperty rights. ranging biotechnologyresearch because they Researchand applicationsinvolving genet- lack the trained scientific personnel, well- icallyengineered organisms may present risks equippedlaboratories and dependablesupplies 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 proceduresas specifiedby appropriateregula- results and exploit them. In view of these tions. Guidelinesfor biotechnologyresearch problems, internationalagencies have a vital and field testing have been formulatedby the role to play by providing technical assistance Organizationfor Economic Cooperationand and by catalyzing joint, cooperative R&D Development(OECD), anda UNEP/UNIDO/ efforts between laboratories and firms in WHO working group has elaborated guide- industrial and developingcountries. lines for field testingapplicable 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 governmentswhen they tween R&D units in developingcountries and formulate local laws. However, adapting counterpartsor industries in industrial coun- foreign guidelines and regulations requires tries. Common to these projects is that both expertise in risk assessmentand communica- sides benefit: the industrial country partner tion. Local scientistsand regulators will have gains access to a raw materialand retains cer- to be trained in these subjects, possibly in tain marketing rights, while the developing countries where regulatory frameworkshave country partner gains expertise, financial been developedand the appropriateskills are backing and regional marketing rights. Some available. of these types of arrangements have been If wisely and correctly employed, marine brokered by internationalagencies, including biotechnologyoffers the tools for increasing the United Nations DevelopmentProgramme high quality food supplies; for the sustain- (UNDP), United Nations IndustrialDevelop- able, environmentallysound exploitationof ment Organization (UNIDO) and World marine natural resources; for deploying Health Organization(WHO). These organiza- bioremediationto 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 continuousmonitoring ing can be minimized by the international of pathogensand pollutantsin coastalwaters. organizationproviding expertise in drawing

1 The marine environment

This chapter has four sections. First, the Oceans can be considered as giant heat vertical and horizontal dimensionsof oceans ponds, collectingand storing energy from the and ocean space are described. Second, the sun. However, because water is an efficient number, variety and uniquenessof marinelife 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- geographiclocation, currentsand 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 Oceansand 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 precipitousdrop in temperatureof 5° the high seas (blue water) and the coastal Celsius (C) or more. Beneath the thermo- margin. The latter is a complexsystem where cline, deeper waters support a profusion of land, sea, fresh water and atmosphereinter- life. However, as light is absorbedby water, phase. Most of the world's humanpopulation the biochemicaland physiologicalcharacteris- lives in the earth's coastal areas, within what tics of many organisms inhabitingthis 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 biologicalproperties. 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 existingbetween the atmosphere and virtually free of pathogens. As is dis- and seabed. Oceanographerscall 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, chemicaland 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 providesoxygen to marine organisms.Oil re- unique mechanisms to survive that most leased in the oceans will drasticallyaffect the inhospitable environment they populate, marine microlayer, principally through slick mechanismsthat may prove extremelyuseful formation. to humanity. 2 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

Marine life the surface that depends on oxygen for ener- gy and carbon for buildingmaterial, animals The oceans that cover 71 percent of the at these extreme marine enviromnentsthrive earth's surface provide a haven for a multi- in the methane-sulfideand hydrogen-sulfide tude of biologicallydiverse 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 symbiosisexisting whale. Science has identified and character- between microorganismsand clams, mussels ized approximately180,000 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 estimatedthat more than 800,000 are methane is picked up and processedby highly yet to be discovered. Unpolluted waters specializedbacteria that live only in the gills everywhere teem with life. It is hard to of this molluscspecies. 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 needsmethane 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- Althoughthe basic buildingblocks 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 endowingthem with biochemical,biophysio- Marine environmental damage logical and genetic characteristics that are and destruction quite differentfrom those exhibitedby terres- trial life. Informationabout 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, scientistsesti- marinelife that provide aquaculturists,fisher- mate that less than 20 percent of terrestrial men and sailors with opportunitiesto earn a organism have been closely investigated;the livelihood, the sports-fishermenwith recre- figure for marine organisms is considerably ation possibilities,and inhabitantsin 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 evolvedelaborate mechanisms fisheries for cod, salmon and sardines have to enable them to flourish in an extreme gone bankruptdue to overfishing;mangroves environmentcharacterized by high pressure, in Asia have been destroyedto make way for high heat and no light. Unlike animal life on prawn farming; coastal waters off Africa and The mardneenvIronment 3

Asia have been polluted by human and densities. If scientistsare able to collect many slaughterhousewastes, 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 interactionsbetween 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 limitationsof direct collectionare: it is labor enormous variety of animals and plants, have intensive; it requires expensive equipment been dynamitedor poisoned. and sophisticatedinstrumentation, including Damage to marine biodiversitycaused by computers,for thoroughanalysis; the samples manmade activities ranges in severity from will always be minute compared to the area barely discernibleto 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 developmentof a natural resource, such as of political, physical or geographicalfactors. 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 governmentsinstitute conserva- detect schoolsof finfish and mollusks, includ- tion and anti-pollutionmeasures 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 negativeeffects identify the detected animals. Acoustic sys- multiply. tems can be used to improve the efficiencyof fisheries by locating and quantifyingmarine Detecting and monitoring living resources. However, sophisticated oceanic events acoustic systems are expensive and need trained personnel for their operation. These Oceanicevents may be detectedand moni- systemsare 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 organismsmay 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 IntergovernmentalOceanographic Com- and to estimate population numbers and mission (IOC) in 1987 launched the Interna- 4 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

tional Mussel Watch experimentto use mus- tures; synthetic aperture radar for wave sels to detect and monitor chlorinatedhydro- studies; microwave radiometers for wind carbons.The preparatoryphase of the experi- speed; and coastal zone color scanners for ment was successful, so a full-scale mussel plankton densities. The remote sensing tech- sentinelprogram was begun in 1990 and will niques, when coordinatedwith surveys done continuethrough 1992 (UNEP 1991). There at ground level, allowsscientists to determine are, however, some limitationsto the sentinel and measure upwellings(the developmentof 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 chemicalor pathogen cost of deploying sophisticatedhardware 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 equipmenton 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 niquesused, for exampleremote sensing, are parameters must interface with an operating sophisticatedand 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- phenomenaas climatechange, 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 productivityof marine organisms, the details and analysis are the same. Aircraft, though, of biogeochemicalcycling, most of the pro- can in addition be used for detailed analogue cesses affecting environmnentalchange, the photography and visual interpretation for transfer of genes in the oceans, and many pursuit of fish and marine mammals (Baker other physical and biologicaloccurrences. As 1991). Remote sensing equipment generates is discussed in the following chapters, ad- data on a wide segment of the electromagne- vanced biotechnologytechniques 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. Remotesensing and the environment: present technology

SATELLITZES LANOSATI-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 le - - 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 AQ 94 SPOT 3,4 P - * 94 SPOT 1 XS , *C) -n 91 JERS-t '- 0 * I'0 92 MOS- 1 U C7.1- 95 AOEOS ' .** * - * * * t

94 RAOARSAT , , ,., 90 KFA 1000 0'''-'' _9D COSMOSi1J.* , - , ,* A 90 Eos - - s 96 SPACESTATION ,*,* *** *,- AIRCRAFr IMPFROVEDMSS *; 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 biologicalagents to providegoods and servic- manufactureon a large scale organic chemi- es." (Bull, Holt and Lilly 1982)Scientific and cals, such as acetone, butanol and ethanol. By engineeringprinciples 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, biochemicaland chemicalengineer- tured via fermentation, a percentage that ing. Researchers in traditional applied fields dropped drastically after World War II be- such as agriculture, aquacultureand 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 essentialcharacteristics 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 substancesthat were difficultto synthesizeby the biosciences and bioengineering.Accord- chemicalmethods, such as steroids, enzymes ingly, biotechnology techniques developed and certain vitamins. before 1973 can be considered "classical' The research carried out by generationsof techniques,while the many genetic engineer- geneticists, biochemists and other bio- ing techniques developed during the last scientistsled to the developmentof the time- twenty years or so can reasonablybe grouped tested techniques of mutation, selection and under the heading of 'advanced' biotechnolo- breeding that drive the conventionalbiotech- gy. For this reason, the primer begins with nologyindustry. Someof 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 biotechnologyoffers for devel- capabilityof Penicilliumnotatum, 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. fermentationin the late 1930s. It is useful to describe a classical industri- Classical biotechnology al biotechnologyproject, one whoseobjective is to develop an antibiotic-producingorgan- While humanshave appliedbiotechnology 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 advancesin the field. The discoveryby Louis sample from a tropical forest or a sponge Pasteur in 1857 that fermentationresults from taken from a coral reef. After screening the action of microorganismsis a fine exam- uncovers an antibiotic-producingorganism, ple of a breakthrough. Capitalizingon find- chemists assay its ability to produce the A primer In biotechnology 7

antibiotic. The natural (or wild) type organ- undesirabletrait appears, a laboriousancillary ism does not usually release the antibioticin breeding process called backcrossingis done sufficiently high concentrationto be useful to eliminateit. for industry. Using low-producingorganisms as parents, the industrial researcherdevelops Advanced biotechnology their progeny to produce greater quantitiesof the antibioticthan the parent. The first step is Advancedbiotechnology is usually consid- usually to expose the parent organism to ered to have begun with the research on mutagenic chemicals, ultra-violetlight or X- animal viruses undertaken by Paul Berg, rays. After treatment, microbiologistspropa- 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 selectedstrain 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 antibioticand that do oped, enablingthe 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 industrialfermenters, 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 undertakesR&D The next significant advance in advanced projects in classical biotechnologywill typi- biotechnologywas in 1975 when the Argen- cally include microbiologists, biochemists, tinian scientistC. Milstein(at the time work- geneticistsand fermentationengineers. ing in the United Kingdom) and the Swiss Scientists use classical biotechnology scientist G. Ko1hlerdiscovered a method for techniquesto 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-producingcereal plant species, or a antibody-producingB-cells are then fused superior antibiotic-secretingmicroorganism 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 simultaneouslyoccur; some or substances: virus, bacteria, a peptide, a could result in unwanted characteristicsap- 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 microorganismmay 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 IOTECHNOLOGY AND DEVELOPNOCOUNTRIES

detect and monitor pathogenic microorgan- ple, strains of the bacterium Escherichiacoil isms and toxic pollutants, and by industry to (E. colt), a microorganismnormally found in capture and recover a specific substancefrom the gastrointestinal tracts of animals, have a solution. had human genes inserted in their genomes. Recombinant DNA and hybridoma con- Industryuses geneticallyengineered E. coli to struction were the first of a series of biotech- produce a range of proteins previouslyunique nology techniquesthat collectively are now to the human being, includinghuman insulin termedgenetic engineering.Genetic engineer- and growth hormone. These achievements ing has been defined by some as the science could not have been accomplishedvia 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 improvingthe yield of an may be employedto develop a novel biologi- amino acid-producing microorganism.Once cal entity (the transgenicorganism); once the researchers identify the gene coding for the transgenicorganism has been realized, scien- amino acid's sequenceit can be defined, then tists employ breeding and selectionto 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 anotherwell-characterized organism, perhaps processes for the large-scaleproduction of the more suitable for industrial purposes. In wanted protein or metabolite.Scientists from either case increasedproduction of the amino many differentdisciplines have contributedto 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 instrumentationengineers results from advanced biotechnology are apply researchfindings to industrialpurposes. 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 recombinationof classical biotechnology has generated for genes that occurs cannot be predetermined. humanity, advanced biotechnology equips Conversely,the genetic engineeringrecombi- bioscientistswith new, powerfultools (UNDP nation in a well-characterizedhost 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 engineeringcan be used edge advancesand techniques are perfected. to combinedesirable 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 approachof mutation,breeding and selec- pharmaceuticalsthat were barely known, or tion. For instance, the developmentof a high were unknown, a few years ago. For exam- yielding amino acid-producing bacterium A primerIn blotechnology 9

through conventionalbreeding and selection Food industry may take several years; the same achievement could be reached within a year through the Genetic engineeringis already making an application of advanced biotechnologytech- impact on the food industry. To illustrate, niques. formerly the only supplyof rennin, an expen- sive substance vital to cheese making, was Biotechnology and the calf rumen. Now genetically engineered developing countries bacteria mass-produceit. Improved availabil- ity of rennin to cheese manufacturersmeans By now, about twenty years after the that they have more dependablesupplies, are discovery of genetic engineering, modem able to improve the quality control of the biotechnologyclearly holds immensepromise 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 traditionalmethods used to produce ferment- shortage, and polluted marine and terrestrial ed foods and beverages also has relevancefor environs (BOSTID 1982; UNIDO 1981). developingcountries. For example, the food Four areas hold especialpromise: agriculture, industry supports research on the genetics of food industry, pharmaceuticals,and industrial lactic acid fermenting bacteria to enrich the chemicals. flavor and nutritive contentof fermentedfood staples. In another example, inediblebiomass 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 discussedbelow, marine biotechnol- diagnostickits based on monoclonalantibod- 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 naturalfood 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 developingcountries by endow- Pharmaceuticals ing them with improved disease resistance, improvingtheir nutritionalvalue, enhancing Industry produces about 80 percent of all their ability to withstanddrought and heat or pharmaceuticalsvia chemicalsynthesis or by to grow in soils that have been damagedby extraction from animal and plant tissues. agrochemicalsor that containhigh concentra- Biologicalmethods, 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 prescriptiondrugs. The proportionof fermen- discussed below, are likely to have uses in tation-produceddrugs will increase dramati- terrestrial agriculture. For example, fish cally in the next ten to twenty years as the genes that code for anti-freezingproteins may new biotechnologytechniques 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 pharmaceuticalsproduced by after freezing and thawing. geneticallyengineered organisms are creating 10 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

new niches in the drug market. To illustrate, going into developing microorganisms that at present approximatelya 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 developingcountries is more probes (see page 48 and following) that en- readily accomplishedthan 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 catastrophicdiseases, and semitropicaldeveloping countries possess such as malaria, hepatitis, human and live- enormous natural resources amenable to stocktrypanosomiasis and schistosomiasis.In development via biotechnology, their deci- fact, two recombinanthepatitis B vaccinesare sionmakers would be remiss in failing to already in use. Others, including vaccines consider building capabilityin this field. against dengue fever and herpes, are likelyto 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 introductionof 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 accidentalor chance temperatures, and promise to be cheaper. recombinationof genes would alter the bacte- rial host, endowingit with undesirablechar- Industrial chemicals acteristics.The public, reflectinguncertainties by scientists, voiced their concerns about Approximately90 percent of the substrates several aspects of biotechnology. Could an used to synthesize chemicals are petroleum- entirely new life form with unknowncharac- based. For the many petroleum-importing teristics be created by researchers? Could countriesthe replacing of chemicalsynthesis otherwise innocuous bacteria accidentally by fermentationprocesses couldhave several become endowed with pathogenic properties beneficial effects. Fermentation occurs at during research and escape from research lower temperaturesand pressuresthan chemi- laboratories? Could new recombinantforms cal reactions,thus savingenergy 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, agriculturaland forestry wastes, and specific Laboratorysafety crops to produce useful starting materials for many industrial chemicals, such as sugars, Most bioscientistsbelieved that the possi- methaneand alcohols.Fermentation processes bility of such events were small, but no hard adapted to use geneticallyengineered 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 additivesin animalhusbandry. 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 conclusionsof the Asilomar conferencewere rDNA experiments presented by local used by the U.S. NationalInstitutes of Health researchers and specifying the conditions (NIH) to formulatea set of guidelines. These under which these should take place. NIH guidelinesfor 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 establishmentquickly experiments and set forth the conditions acceptedthe NIH guidelines.Simultaneously, under which research could take place. the United Kingdom establishedthe 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 guidelinesthat were adapted from guidelines work surfaces, wearing laboratory existingelsewhere. 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 assessmentexperiments tory practices that any well-run hospital to try deliberatelyto create pathogens.These clinical or research laboratory would confirmed that the possibilityof 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-containedunits 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 scientistsusing 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 conventionalmeans. 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 scientificknowledge. 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 conductedthroughout the Institutional Biosafety Committee (IBC), world in the agricultural, biological, indus- 12 MARINEBIOTECHNOLOGYANDDEVELOPING 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 engineeringtechniques are safe. means. In fact, biosafetyregulatory programs There are three major reasons for the in the United States, the European Commun- safetyof rDNA research. First, the successful ity and elsewhere assess inanimateproducts 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 conventionalresearch 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 considerableextent 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- aimedspecifically at inanimatebiotechnology ulating only one or a few genes is minute. products. Second, genes include regulatory DNA se- quences, called operons, that control in a Deliberaterelease positiveor negativeway the expressionof 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-associatedgenes couldbe identifyand quantifythe risks associatedwith accidentally created. Third, the insertion of introducinggenetically engineered organisms alien genes in a microorganismusually 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 organismsor environmentor 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 accumulatedproving the safety of non-targetorganisms. biotechnologyresearch, 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 situationsand producedby geneticallyengineered organisms concludedthat there are three essentialcrite- and the deliberate release of genetically ria for evaluating the risks associatedwith a engineeredorganisms into the environment. proposed release (NRC 1989):

Product safety * Are we familiar with the properties of the organism and the environmentinto which Industry marketsnumerous products made it may be introduced? by geneticallyengineered organisms, includ- ing human insulin, human growth hormone, * Can we confine or control the organism various animal growth hormones, interferons effectively? A primerIn biotechnology13

What are the probable effects on the called Biotrack Information System. As of environmentshould the introducedorgan- 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 managingthe field testing of geneticallyengineered 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 geneticallyengi- situations and would apply equally to any neered organisms in the field are dealt with proposed release of genetically engineered on a case-by-casebasis. In general, however, organisms into the marine or freshwater the process is as follows. For any proposal, enviromnents.It is illustrativeto considerthe a thorough environmentalimpact assessment one field test involvingtransgenic fish in light is conducted.The assessmentaddresses health of these criteria. To do this we need to re- and safety concerns by considering both view the events that preceded the decisionto direct and indirect effects stemmingfrom the allowthe test and to scrutinize the-conditions proposed release. It must convincinglyshow under which this test is being performed. that the proposed release would probably not The fish in question is a carp containinga 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-targetanimals or plants; for University (Alabama) (Chen and Powers example, because it possessedcharacteristics 1990). At the end of 1989, the team request- such as enhanced fitness, increasedpathoge- 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 affectsthe reproductivecapacity 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 opportunityto scrutinizethe environ- offspring will develop and behave as do the mental impact assessmentreport and to com- offspring from "normal" carp. This research ment on it. The regulating agency must take would be generally useful for improvingthe these comments, as well as statements made carp's genetic characteristicsfor 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 NationalWildlife Federation, geneticallyengineered organisms, including a on grounds that carp have a significantpoten- transgenic fish (see below). No negative tial to damageinsects, 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 initialproposal was remandedand ing, at least in the short term (Miller, Burris its drafters were asked to provide more and and Vidaver 1991). (Field tests throughout better informationabout possibleenvironmen- the world are recorded in an OECD database tal impacts. 14 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

About six months later, the principal resemblestesting in a closed systemthan true investigatorssubmitted a redrafted proposal field testing. Nevertheless, it probably is a to the USDA. It asked for permissionto raise model for the initial field testing of any 50,000 fry that had been spawned from nine aquatic animalor plant. As such, it is elabo- transgenic carp in ten outdoor pools. After rate and expensive,hardly an undertakingfor three months, the number of fry would be most developingcountries. (Safety consider- reduced to 300 per pond; these would be ations as they relate specifically to marine marked for identificationand studied for the biotechnologyare discussed at length in the next fifteen months. The fish would then be next section.) destroyed,before they reached sexualmaturi- ty. The ponds stocking the fish would be Internationalguidelines well-protectedand would have no connection to any other waterways. During the last few years, safety aspects After some months of public hearings and of biotechnologyhas become a subject of deliberations, the USDA determined that the interest and concern by policymakerson the " .experiment with transgenic carp presents internationallevel. 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 developingcountries have been elaboratedby 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 conditionsof the field testingof 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 studiedand 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 physicalproperties, except that bearing on indicate a new trend; that is, governments growth. Whetherthe 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 geneticallyengi- closed, artificialsystem, the environmentinto neered microbe, plant or fish after release or which the transgeniccarp 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 internationalagen- deliberate, criminal attempt was made to cies are helpful, a difficultproblem still faces release them, the conditionsunder which the governmentsof developingcountries, because testing of the transgenic carp is taking place expertise in risk assessmentand communica- precludes the possibilityof their escaping. So tion is necessary in order for them to adapt certainly the second criterion is satisfied;the foreign guidelinesand regulationsto 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 conditionspreclude persis- countries. Local scientists and regulators tence or spread. should therefore be encouraged to acquire The field testing of the transgeniccarp 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, advancedbiotechnology formulatepolicies for biotechnologyresearch, 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 Engineeringand 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 activitiesconsist- advances, questionsinevitably arise about its ed of courses offered in Triesteto 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 biotechnologyand certainmarine "Genetically Modified Organisms for the applications. Accordingly, the next sections: 1990s," with fifty participants.Topics includ- (1) considerbiosafety issues that biotechnolo- ed biological risk assessment,containment of gy in general has generatedand analyzethem genetically-modifiedorganisms in the labora- in terms of their relevance to marine biotech- tory and the field, recommendedprocedures nology; (2) analyze special characteristicsof for safe laboratory and industrial practices, the marine environment that bear on bio- transgenic animals, and analysis of existing safety; (3) scrutinize introductionsof exotic biosafety legislation. The objective of these aquatic organismsinto 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 assessmentand 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 circumstancesand conditions. It bears mentioningat this point that there In the preceding sections it was seen that is some concern that the biotechnologyindus- biotechnology research raises one set of try will be a threat to producers of natural concerns, biotechnology products another. products. For example, in industrialcountries Each requires reflection to gain a perspective cell culture systems that will mass produce on the possiblebiohazards 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 displacedby 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 stringencyof conditionsunder 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 importantsocio-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 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

dures to ensure the safety of themselves, environment pose a hazard to existing life other workers, and the surroundingcommuni- forms or the environment? ty. Conversely, research involving a non- Concerning the first question, most inani- pathogenusually requires no more than good mate productsfrom biotechnologyare known laboratory practices. It is probable that over chemicals or compounds produced via fer- 95 percent of all biotechnologyresearch is mentation. However, rDNA technologyhas being done under the conditions defined by enabledthe mass productionof 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 chemicaland physio- which in turn have generated new marketing logical characteristics, the conditions under niches, changedthe way we regard intellectu- which scientists conduct research are analo- al property and, at times, created ethical gous in marine biotechnologyand 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 governmentaland intergovernmentalagencies biology employs the same techniquesas 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 equipmentand from that of conventionallyproduced prod- reagents. Furthermore, having researchers ucts; the same criteria of safety and efficacy investigatinga 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 issuesposed by marine biotechnol- intended for animal or plant use, or is a ogy research performed in the closed system nonconsumablecommodity, 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 biotechnologyresearch. For these exacting procedures, includingclinical phas- reasons, the voluntary guidelinesthat 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, regulationsthat specify testingof the product scientists, public advocates and regulators and that establish mechanismsfor monitoring seem to be in agreement on this point. the testing procedures. The chemicalstructures and other charac- BIOTECHNOLOGYPRODUCTS AND BIO- teristics of existing products from marine sAFETY.Advanced biotechnology research biotechnology are the same or similar to can producetwo types of products-inanimate known compoundsfrom the terrestrial envi- products and geneticallyaltered living organ- ronment, although some have unique struc- isms. Each poses important questions, such tures. As more organisms from extreme as: Do inanimateproducts 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 conventionallypro- properties, will be found. However, if the duced products? Would the deliberaterelease experience of terrestrial biotechnology is a of geneticallyengineered 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 geologicalbarriers preventingthe create a novel situation, or uncommonhaz- 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 continuityof 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 characteristicfavors, if not lished procedureswould unravel the chemical the dispersal of whole organisms, of genetic structure of the new compound, explain its material.This characteristicis 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 recombinantkilled-vaccine is developed, viability of immersed organisms and, at its field testing would most likely follow times, parts of organisms by preventing establishedanimal vaccine testing procedures; desiccation and deflecting deadly ultraviolet testing would be satisfactorily monitored by light. existingnational 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 conventionalor advanced come into direct contact with other organisms biotechniquesare appropriate for use in the and diverse suspendedmatter, creatingpossi- 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 exchangegenetic 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 geneticallymodified organisms in the marine efficientlybetween two species in the family environment. Nevertheless, such a proposal Enterobacteriaceae,say E. coli and Salmo- could soon be submittedso 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 exoticbacterium couldpossibly thus pass teristics of the marine environmentmust first on the alien gene to a wild bacterium through be examinedbecause they will determinehow conjugation. Little is actually known about marine organismsand discrete geneticmateri- conjugation among marine bacterial species, al (for example, plasmids and naked DNA) but it is reasonable to believe that dispersal disperse. by conjugationwould be more likely in water populatedby high numbersof bacteria due to Special characteristicsof the marine contaminationby sewage and human wastes environmentthat bear on biosafety than in blue ocean water.

One of the most important characteristics TRANSDUCI-ION.The second mechanismis of oceans is that they, like the atmosphere, transduction, where a vector transfers the are continuous-there are therefore no geo- genetic materialfrom one cell to another. For 18 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

example, the bacterialspecies Agrobacterium Biologicalinvasion is "the arrival, estab- tumefaciensis a useful vector for transferring lishment, and subsequentdiffusion of species genes into many plant species; simple virus- in a communityin which they did not previ- es, called bacteriophages(or phage for short), ously exist in historicaltimes. "(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 naturalmechanisms. This phenomenonhas recentresearch demonstratesthat 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 createsproblems for risk assessors;problems ocean of viruses 1990). The role of viruses in that cannot be resolved until much basic the marine environmentis 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, Introductionis the accidentalor deliberate microalgae,plankton and other organisms. dispersal of organismsthrough human activi- ties (Carlton 1989). With the introductionof 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 organismsfrom one place to another occurs in the laboratory, wherethe researcher in their ballasts, encrusted on their hulls, and creates the chemical environment (media) bored in their wooden hulls. The openingsof conducive to the reaction in a contained interoceanicand interlake canals have given vessel. Transformationsappear 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 molluskslong distancesfrom fishing grounds the dispersal of genes via transformation in to market places. Pathogensthat afflict these the media that is ocean water. fishery products have been carried along (Carlton 1989). Marine species numberingin The dispersal of marine organisms the thousands have thus been moved across and their sequel the globe in innumerablepatterns since trans- oceanic trading commenced. Past examples of dispersals of marine Besidesaccidental 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 engineeredmarine organism, shouldone 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 primerIn biotechnology 19

result of the breeding of new, more desirable larva-eating fish may eat eggs and larva of fish, shellfishand crustaceanstrains. Thus, in other fish; the grass carp (Ctenopharyngodon the 1950s and 1960s there were large-scale idella) transmits a cestode-causedfish dis- deliberate introductionsof fish and shellfish ease; the Pacificseaweed Sargassum muticum throughout developing countries. Introduc- was inadvertently introduced with C. gigas tions includedthe African 7ilapiato 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-eatingfish Gambusiaaffinis and Lebis- widely introducedshrimp 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 infectioushypodermal 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-scaleintro- 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 (Crassostreagigas) 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 developingcountries. waters; the pink salmon (0. gorbuscha) to One is the example of the introductionof the the Arctic Sea coast of Russia; a shrimp Nile perch Lates niloticus into Lake Victoria species from Panama (Penaeusstylirostris) to during the 1950s. The fish establisheditself 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 introducedfrom the Philippinesto 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 deliberateintroductions have practical way of preserving perch harvests benefittedlocal populationsand improvedthe proved to be smoking, which demandedgreat economiesof countries. For example, France quantitiesof 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 introductionof the Nile perch was destructive 12,000 tons from Lake Kariba; and Sri to aquatic and terrestrial biodiversity, while Lanka's entire inland aquacultureproduction the economic return from the introduction of 32,000 tons consistsof introducedfish and could not be sustained. crustaceans (Sindermann 1986; Welcomme The second case is the introduction in 1986). The value of malaria larva-eatingfish 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 introducedspecies 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 marketnever developed, 20 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

however, and local consumptionis low. The * New diseases may be introduced along snail meanwhile thrives in the rice fields, with the deliberate introductionof a stock. where it turned into a pest, attacking newly transplantedrice 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, (Welcomme1986). which is resistant to pesticides and other control measures. The International Centre * Once an introducedorganism colonizesa for Living Aquatic Resources Management locale, it may be impossibleto 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 engenderedthrough- farming methodsto control the snail (Lessons out the world, governmentsand international 1992). agencieshave sought to prevent future prob- The foregoing indicates that the extent of lems and to alleviate existing ones through long-termdamage from accidentalor deliber- the adoptionand implementationof codes and ate introductionsof exotic animal speciesinto 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 AdverseEffects Arisingfrom Introductions nor is it possible to determine with certainty and Transferof Marine Species adopted by whether the benefits stemmingfrom deliber- the InternationalCouncil 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 followingproblems may result from introduc- statements and conventions on the subject tions: have been made by the American Fisheries Society (1973), the U.N. Conferenceon the * An introduced animal may disrupt local Law of the Sea (1982), the Councilof Europe fauna through competition or predation. In (1984), FAO's European Inland Fisheries the worst case, the introductionof an exotic AdvisoryCommission (1984) and the Interna- species may lead to the extinctionof the wild tional Union for Conservationof Nature and species. Natural Resources (1987). The overriding objective of these codes and statements is to * Genetic degradationof the host stock may direct concerted international action toward result from its introductioninto 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 implementationof 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- populationidentity. In other words, when an riding strategy is for U.N. agencies and introduced species breeds with the wild nongovernmentalorganizations to educatethe species inhabiting a locale, the adaptions for public, policymakersand 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 primerin 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 introductionsand to delineatethe condi- areas of marine biotechnology. tions under which authorized introductions The difference, then, is in field testing of may proceed. Another strategy is suggested geneticallyengineered organisms. As noted, for larger, industrialcountries. It emphasizes USDA has given permission for the field regionalapproaches to controllingthe 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 assumptionthat and the regulator faces special problems not risks from introductionsare 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 reductionincludes the thoroughstudy cannot be guaranteed because of the three of the organism proposed for introductionin characteristics of the marine envi- its native habitat; considerationof developing ronment-the continuity of the oceans, the native stocks as an alternativeto introducing perpetual motion of the water and suspended a new stock; and establishmentof mecha- particles, and the potential for gene disper- nisms for monitoring the introduced stock sion via unfamiliar biological mechanisms. continuously.It is particularly importantthat 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 includingclarifying ecological considerations escapes, the consequencesare incalculable. (such as competitionand 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- conditionsof the field test itself, it is perhaps ioral consideration (including interactions more useful for our purposes to speculateon between the introduced and native species); the consequencesof a successfultest; that is, and pathologicalconsiderations (including the if the tests convincinglydemonstrate that the possibility that the introduced species will standard culturing of the transgenic carp is carry along with it new infectiousdiseases) more cost-effective than culturing present (Sindermann 1986). stocks, what then? One conceivablescenario is that someone would try to take advantage Comparisonof terrestrialand of the higherperforming characteristicsof the marine biotechnology transgenicfish by intensively culturingthem in cages or pens emplaced in a lake or a Does marine biotechnologypose different river. The possibilityof some transgenicfish safety and regulatory issues from terrestrial escaping would be high. What would be the biotechnology? As was discussedpreviously, consequencesof an escape? marine biotechnology research does not, Referring to the six possibleproblems that neither does inanimateproducts from marine past introductions have caused, the conse- 22 MARINEBIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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 organismsthat 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 completelydiscount 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 consequencesequel, 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 dangersof genetic manipulations added safety, triploid inductioncan be com- shouldbe recognized,and biotechnolo- bined with further treatmentthat producesan 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 justificationfor maintainingthat Kapuscinskipoints out that even if only diversity. The release of individuals sterile transgenicfish are cultured, some risk with artificially composed genetic remains because of the necessity to maintain makeups into wild populationsof 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 educateeveryone who works the competitiveinteractions with other with them of the ecological problems that species, destabilizingnatural 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 transgenicfish may be the first candidate The field testingof a transgenicfish 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 technicalones. Accord- bacteria (for bioremediation)and microalgae ing to Dr. A. Kapuscinski,who works for the (for increasedproduction of food additives), Department of Fisheries and Wildlife, Uni- they rather than multi-celled animals should versity of Minnesota, technicalbarriers must be considered as the primary candidates for be overcomeand environmentalrisks reduced the first marine field testing. Further support before transgenicfish can be cultured(Kapus- for this contentioncomes from the report that cinski 1990). The technical barriers relate to the firm Envirogen Inc. in New Jersey is the cost-effectivetransfer 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 identificationof transformedindividuals 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 proposalwill 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 transgenicfish 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 methodologyfor field testing in ability outside the laboratory, ability of the the oceans has been developed. Without organism to degrade trichlorethyleneunder doubt, for the present it is more difficult to various conditions and in the presence of evaluate and determinethe possible effects of sundry chemicals, and whether synergism is the field testing in the marine environmentof possible between the tested organisms and transgenicmarine animals, plants and micro- other microorganisms. Such responsible organismsthan in the terrestrial environment. testing would not likely endanger man or To concludethis chapter, the expectations environment. and problems of marine species field testing The field testing of a transgenic bacteri- today are approximatelythe same as it was um, and other microorganisms,should for the for terrestrial and/or atmosphericfield testing present not be done in an open system. For when these activitieswere commencingsome one, little is known about the dispersal of ten years ago. Unlikeformer times, however, whole microorganismsin 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 functioningand are thus better prepared to prepare compre- efficiency of the three mechanismsfor 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 methodologiesadapted for the marine envi- carried by the transgenic microorganism ronment, design safe test protocols, and to would disperse, the frequency of possible institute efficientmechanisms 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, comprehensiveregula- marine environmentshould be deferred until tory regime in place that will ensure proper research in biological oceanography,micro- precautionsare taken, while preventingpoor- bial ecology and environmental toxicology ly planned and executedfield tests from being have clarified the details of the mechanisms undertaken. 3 Marine biotechnology and its sub-areas

Althoughmuch 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 Althoughthousands of projects have been In a strict sense, "...any scientific investi- or are being undertaken that can be consid- gation that focusses on marine organismsand ered as marine biotechnology R&D, it is that utilizes new cell, protein and nucleicacid possible and convenientto 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. believedtoo narrow by many researcherswho hold that a host of different R&D activities Aquaculture rightfully are sub-areasof marine biotechnol- ogy. Taking into account this lack of consen- Aquaculture is a general term that refers sus on what marine biotechnologyconstitutes to the husbandryof 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 approximately10 million tons in 1985 to 14 employ biotechnology techniques. Thus, a million tons today, and estimated at 22 mil- broad definitionis analogousto the definition lion tons by 2000. If these projections hold of biotechnologygiven in Chapter 2 (Bull, true, FAO estimates that by the end of the Holt and Lilly 1982). Marine biotechnology century aquacultureproducts will accountfor 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 AppendixC). mately99.5 million tons in 1989 accordingto When biotechnologytechniques are used FAO (1989 FAO statistics 1992), which is for research in certain appliedfields, such as very near to its estimated maximum sustain- aquaculture, fisheries and natural marine able yield of about 100 milliontons per year. products, they are includedunder 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 discussedbelow, 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 contaminantsthat compete with crop plants fish exceeds 1 kilogram of fish per 57 cubic for food and sunlight (Gellenbeckand Chap- decimeters of water or 1400 kilograms of man 1983). Further, wastes from seafood shrimp per 0.4 hectares of water (McCoy processingplants associatedwith aquaculture 1990). For example, in culturing prawns in can create disposal problems; wastes usually the Philippines,the stockingrate for the giant are 20-25 percent for finfish and 80-85 tiger prawn, Penaeus monodon, in ponds percentfor shellfish (TechnicalQ&A 1991b). where traditionaltechniques 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, conditionscreated 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- ornamentalfish is a relativelynew but rapidly crease yields from aquacultureor to enhance growing area of aquaculture. the quality of its products. In particular, Mariculture is that subset of aquaculture biotechnologyoffers new methodsto improve that is practiced in salt water. The term the heath status of cultured organisms (see mariculture is specificand 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. discussionof specific marine biotechnology The practice of aquacultureconsists of the applicationsto these organismsfollow. applicationof a set of low-technologyendeav- ors pertaining to the breeding, propagation, GROWrHAND REGULATIONOF 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 problemsencountered 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 aquaculturistneeds to know the composition was the extractionof the hGH from the gland and chemical characteristics of the soil in difficult, but also approximately5000 of the which ponds will be constructed,the quantity minute glands were required in order to and qualityof the water in whichanimals will produce one gram of the substance. After grow, and the ready availability of animal techniqueswere perfected for the large-scale feed. Further, incorrectly sited ponds may manufactureof hGH produced by genetically severely damagethe nearby environment.For engineeredbacteria, or recombinanthGH for instance, pond constructionhas often entailed short, sufficient quantities have become the destruction of mangroves; sweet water availableto treat all who need it and to fully used in hatcheries may deplete underground supplymedical researchers. aquifers of fresh water; and untreated efflu- The developmentalprocess that resulted in ents and wastes from farms and hatcheries recombinant hGH has been duplicated in 26 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

regard to animal growth hormones. Thus, gene was transmitted to subsequent genera- beginning in the early 1970s, extraction tions of progeny. techniqueswere developedto recover various Much effort in industrializedcountries is growth hormones from their respective ani- being devoted to producing transgenic fish. mal pituitary glands, including several fish Fish are good candidatesfor studyfor 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 hormoneswere availableto ideal subjects for genetic manipulation; the researchers. After techniqueswere 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 geneticallyaltered may be Biotechnology Institute's Center of Marine more valuable commercially than their un- Biotechnologygenetically engineered E. coli treated relatives (Liu and others 1990). to produce rainbow trout growth hormone There are three stages to developing a (tGH). In subsequentexperiments, the geneti- transgenicfish. 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, (Agellonand others 1988).Other experiments the foreign gene that is to be inserted into the involvingthe injectionof recombinantsalmon new host is cloned to produce a large number GH into rainbow trout and recombinanttuna of that gene. Then, a gene constructis 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). Althoughthe 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-implantedinto 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 developedto 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 spectacularmanifestation of through the production of GH. Proof of appliedmolecular genetics-the development expression comes from the analysis of the of transgenicanimals. These are animalsinto 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 includethe 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 techniqueshave been particularly successful, been successfully transformed (Chen and includingseveral species of fish, the foreign Powers 1990). Marine biotechnology and Its sub-areas 27

Third, the new trait has to be transmitted pleuronectesyokohamae, which inhabits the to subsequent generations of progeny. The YellowSea. Afterhaving isolatedand charac- trait in progeny is detected in the same man- terized the gene coding for this protein, these ner as in the foregoing step. Consideringthe scientists were able to synthesize this gene ten fish wherein expressionwas 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 (Yaoqingand Xiongfeng 1990). 1990). In addition to the safety concerns that The progeny from one of the two success- surround the developmentof transgenic fish ful experiments are now being field tested. (and other transgenic organisms), difficult Researchersat AuburnUniversity in Alabama scientificand technical problemswill 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- (Kapuscinski1990). 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 effectivenessof 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, salmonidsand trout, do not have this GASTROPODS AND CRUSTACEANS. The major protein. The susceptibilityof salmonto death groups of bivalves are clams, oysters, mus- by freezing has, for instance, prevented sels and cockles; gastropodsinclude abalone; salmon aquaculture in Atlantic Canada (Cut- while crustaceansinclude shrimp, crabs and ler, Saleem and Georges 1989). Inserting the lobsters. Oysters are considereda delicacy by gene coding for the anti-freezeprotein 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 opportunityto 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 spawningof 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 Althoughthe 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 maturitywhen they metamor- against freezing (Chen and Powers 1990). phose into juvenile oysters (spat). Spatusual- Work to improve expressionof the genes is ly set on cultch, where they harden until final proceeding rapidly, however. Antifreeze maturation, whichis called "grow-out." Har- proteins are also being studied in China, dening and grow-out can be induced to take where researchers at the Instituteof Genetics, place on the seabed or in floatingrafts. 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-lessoyster has two aquacultureof oysters. The applicationof one advantagesover 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 chromosomesinstead of the normal two, come manifest. Second, the culinary quality were first developed by researchers at the of the cuitch-lessoyster 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-lessoyster species' qualities, an indus- done by scientistsin 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 chromosomesin 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 containingone where they attain their adult form and remain chromosomeset (Allen 1988). the rest of their lives. The settlingphenomena After oysters was deemedsafe 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 aquaculturistsin 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 involvingabalone, this group showed duringthe summer and becomesless flavorful that spawning (the release of fertilized eggs because reproductivetissue 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 valuableto dins is controlled by the rather common aquafarmersbecause 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 amountsof hydrogen perox- represented about 50 percent of the U.S. ide to the seawatersurrounding 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 metamorphosisof the larvae depend on disease is a serious problem to aquaculturists. the recognition by the larvae of specific Oyster productionin the ChesapeakeBay, 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 pollutionand 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 Marylandto devel- the algae or nearby surface. The chemical op and propagate cultch-lessoysters (that is, that induces settling was found to be a pep- Marine blotechnologyand Its sub-areas 29

tide (a short chain of amino acids), similar to Biotechnology-relatedresearch on crusta- a neurotransmitterchemical found in animals cean species lags behind that being done on called GABA (gamma-aminobutyricacid). 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 GROWITHAND REGULATIONOF ALGAE. giant clams, mussels, oysters, scallops and Algae are nonvascular, photosyntheticplants other bivalves. The aggregateof research has that contain chlorophyll (Robinson 1985). led to an explosion in the knowledge about They may range in size and complexityfrom the molecular mechanismsof sensory recep- the microscopic, single-cell to the 70-meter tors, gene expressionand cellular responseto giant kelp Macrocystis.It is importantto 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 additionalinput in driving up prices and stimulatingoverfishing 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 Californiaabalone has become fuels. a rare treat purchased at a very high price. Algae are generally classifiedaccording to To meet the marketdemand, 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 hydrogenperoxide 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 techniqueshave been ceae, or brown algae (25 percent); and Ctlo- establishedin California; abaloneand 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 commercialimportance 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 Crassostreavirginica aquaculture. The total world production of (Colwell 1986). This accomplishmentsets 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 IOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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 importantfood staple; in 1986 9 x 109 chemicals. Two algal aquaculturetechniques 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 industrialimportance of of the coastal zone. One outplanting tech- algae is that they contain a group of chemi- nique consistsof suspendingropes 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 worldwidedemand 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) monolinestretched along the seabed.Harvest- and pharmaceuticalindustry (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). Outplantingis for the substancestood at 20,000 tons (Rich- practiced extensivelyin 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 secondaquaculture technique is grow- agars (from red algae) was 6,700 tons; this ing algae in a closed system. In this method, quantityis insufficientto 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 pharmaceuticalindustry (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 industryprocesses. 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; dissolvingthe raw hydro- requires the additionof nitrogenousfertilizer. colloidswith alkali; clarifyingand 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 inclementweather. Closed systemsare (Mabeau, Valiat and Brault 1990). Algae used by Europeans, Filipinos and Taiwanese processing is a low to medium technology to cultivatespecies of red algae. endeavorto whichbiotechnology would seem Algae have several uses. Bulk macroalgae to have little application. However, biotech- is used by Asian farmers as fertilizer and to nologytechniques 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 abundantlyavailable. To illustrate, the cost example, researchersat NortheasternUniver- of efficiently producing one kilogram of sity in Boston (USA), are developinghybrid Chlorellais about $10, while Spirulina costs strains of agar-producingmacroalgae 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 microalgaehas 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 productioncosts of microalgaeis dramati- costs $250-$40,000 per kilogram (Renn cally lowered, regulators will have to deter- 1991). Other strains could be developedthat mine whether dried microalgaeis safe when containincreased amounts of specialtychemi- consumed in large quantities. Appropriate cals and pharmaceuticals.It is interestingto testing will have to be done to settle this note that while 80 percent of the macroalgae question. destined for commerceoriginate in the devel- Beyond food, microalgae can be used to oping countriesof the Asia-Pacificregion, 90 manufacturesubstances of interest and value percent of the colloid industry is located in to industry. For example, food industry and West Europe, Japan and Korea (Ruyingand 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 biotechnologyis the work on system has been developedby Microbio Re- macroscopic alga at the University of North sources, San Francisco (USA), consistingof Carolina Center for Marine Science Re- large open outdoor tanks or ponds populated search. Researchers are using cell culture by Dunaliella, which synthesizelarge quanti- techniquesto enhancethe growth characteris- ties of the VitaminA precursorbeta carotene. tics or red alga for the purpose of increasing Similarly, the Israeli firm Koor Foods is this species' ability to produce agar. An developingDunaliella bardawilli, a speciesof interesting aside is that parallel work, using microalgae found in the Sinai peninsula, for the same techniques, is proceedingon angio- glyceroland beta caroteneproduction. Anoth- sperm (a plant, such as eelgrass, that grows er Israeli group at the Ben Gurion University well in shallow waters). The objectiveof this is investigatingthe possibility of using Por- work is to enhancethe 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-likepolysaccharide in large out- dunes in place and to stabilize mud flats door salt water ponds (Weiner 1985). (CMSR 1990). Althoughthe classicaltechniques of muta- MicroalgalR&D has been focussedmostly tion, selection and breeding are most com- on four species; CZlorella,Dunaliella, Scene- mon to microalgae R&D, advancedbiotech- desmus and Spirulina. The approach used is nology techniquesare being used to develop similar to that in industrial microbiology, strains that will grow faster, be more resistant namelymutagenesis and selection of mutants to disease, and better adaptedfor mass propa- with desired traits. The aim of Chlorellaand 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 remainedelusive certain problems have to be overcome in 32 MARINEBIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

microalgaebiotechnology. 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, shalloweror the new host has to be monitoredas does its deeper water than is now possible ability to express the desired product. Third, * possess better nutritional characteristics the stability of the introducedgene 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 continueto function as expect- ed. In addition, when the microalgal cell Marine animal health divides, the gene must be replicated and passed on in subsequentgenerations of proge- Bacterial, fungal, protozoan and viral ny. Although encouragingprogress is being infectious diseases are widespread among reported, scientists have not solved these naturalfish populations.But animalsraised in three problems (Brown, Dunahay and Jarvis intensive aquaculture are especiallyvulnera- 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 mariculturethe major pathogen search Institute in Colorado (USA) is apply- is Vibrio. Pollutedwaters favor the develop- ing rDNA to isolate genes that regulate lipid ment of fungal diseases; the most common biosynthesisand to introducethese genes into agent is Saprolegnia(Shariff and Subasinghe microalgae that have potential for outdoor 1990). Two types of viral diseasesare partic- mass culture (Brown, Dunahay and Jarvis ularly deadly to aquaculturedsalmon-infec- 1989), while the Australian company Wes- tious pancreaticnecrosis and infectioushema- farmers Algal Biotechnology's success in topoietic necrosis (IHN). Similar diseases developing microalgaefor the production of afflict catfish, flounder,menhaden and striped beta carotene and animal feed. Other micro- bass (Klausner 1985). Protozoans, mainly algaeproduction 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 UlcerativeSyndrome is pharmaceuticals and polysaccharides(Bene- a widespread disease afflicting fresh and mann 1989). brackishwater fish; the causativeorganism is To sum up, whether macroalgaeor micro- unknown. Bacterial,fungal and viral diseases algae, the advancedtechniques of biotechnol- also afflict bivalves and crustaceans. ogy could be employedto improve the algae In general, waterbornebacterial and viral itself, making it more useful to industry. pathogens are attenuated in well-managed Theoretically,these improvementsmay devel- aquacultureponds 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 Marineblotechnology and Its sub-wreas33

ponds and tanks holding the diseased fish. aquaculturists.Only two efficaciousvaccines The quantitiesof antibioticsused in aquacul- are, however, available for fish and one for ture is large, for example, in Norway, which lobster; none is availableto prevent diseases is the world's largest producer of aqua- afflicting shrimp and shellfish. cultured salmon, aquaculturistsused 17,000 Whether a scientistis attemptingto 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- commonlyused in aquacultureinclude amino- cines; attenuated, killed and biosynthetic glycosides, beta lactams, tetracyclines, vaccines. Attenuated vaccines consist of macrolides and chloramphenicol. Most of pathogenic organismsthat have been treated them are also used to treat infectiousdiseases so they are no longer infective, but are still of humans. The employmentof antibioticsin able to elicit an antibody reaction in the aquaculture poses potentially serious public vaccinated host. The attenuated vaccine is health and environmental problems since somewhatmore 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- causingtyphoid fever, choleraand dysentery. genic organism. Although safe, this vaccine Further, if improperly used, residues of type may elicit a weaker antibodyresponse in antibiotics may taint the flesh of harvested the host than do the others. The third vaccine fish. Proper managementpractices can lessen type is one developed via genetic engineer- risks associated with antibiotic usage in ing. Briefly, a protein constitutingthe patho- aquaculture, but cannoteliminate 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 quantitiesof the protein. The afflictingtheir 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, affectingmarine animalsare 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 characterizeinfectious agents afflicting metric tons in 1988 and 30,000 tons in 1991 marine animals and to clarify the intricate (Record year 1992), largelydue 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 concentrationof the vac- 34 MARINEBIOTECHNOLOGYANDDEVELOPING 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 immersionmethod is cost-effective,but some levels of the protein they code for have been vaccines will not work when administered expressed (Englekingand Leong 1991). The this way. Vaccinating fish individually is a next step is to scale up the manufacture of manpower-intensiveexercise; 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 relativelyvaluable (for example,food-fish Substancesother than vaccines may have that are near to market size and fish valued protective functions.For example, an extract by collectors). It bears mentioningthat a new from the shellfish Ecteinascidia turbinada method for administering vaccines (and protects eel from Aeromonasinfection and in therapeutics)to fish is being tested. It utilizes generalenhances the immunologicaldefenses a combinationof immersionand ultrasound. of blue crab, crayfish and prawn (Colwell Fish immersed in tanks that are exposed to 1986). The Phillips Petroleum Company ultrasoundfor 10-15 minutesrecord 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 administereddrug (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 vaccineagainst 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 massivedie-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 (Klausner1985). Diagnosingthese diseasesat 1990). In view of the damage this disease an early stage may help in limiting loss of causes, it is understandablethat much effort fish stock, as carriers of the viruses causing is being put into investigatingthe IHN virus the diseasescan be identified and eliminated, and developing a vaccine against it. In fact, limiting the spread of diseases. In addition, prototypesof the three types of vaccineshave 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 naturalproducts (a conventional type killed vaccine) proved not useful when administeredin water. The Natural resource chemists have for over second, the attenuatedtype, was effectivevia 100 years been screening the world's organ- water borne inoculation,but questionsregard- 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 recombinantkilled type 20,000 chemicalsfrom naturalproducts have vaccine, shows most promise in terms of been characterized and the annual sales of Marlne blotechnologyand its sub-areas 35

pharmaceuticalsderived 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 pharmaceuticalindustry 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 environmentare 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 substancesare utility to humans because they are too toxic, discovered every year. unstable, or possess other undesirablecharac- 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 metabolitesand cofactors, whichare essential metabolitesthat possess antibioticproperties, 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 investigationof 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-shelledand slow moving, appearsvulner- organisms, such as genus, species, or even able to predators. Yet, it is but rarely at- strain, often produce substances distinct to tacked. Upon investigationit was found that that group. Since secondary metabolites are the animal secretes an antibiotic substance, by far the most important group of useful whichscientists eventually isolated and identi- chemicals produced by marine organisms, fied, then named mimosamycin (Scheuer and since their evaluationis in fact the objec- 1990). Mimosamycinbelongs to a chemical tive of marine natural product research, group named isoquinolinequinones,which discussionhere 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 mimosamycinis 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 metabolitesas insecticidesand herbicides. ANI-viRAL. As mentioned, most antibiotics cannot be used in animals, includinghumans, 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 competitionfrom other organisms. Some are nonselective in that they interfere with uni- able to gain an advantageby 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 dividingand growing 36 MARINEBIOTECHNOLOGYAND 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 compoundsor differentmicroorganisms treating tumors. Two such agents are now in that live in a commensalor symbiotic rela- clinicaltrials 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 anaerobicbacteria, cyanobacteria,dinoflagel- relationshipwith a yellow sponge found only lates, heterotrophicbacteria, and microalgae in the Gulf of California, Mexico. The sec- inhabit them. When tested, some of these ond is DidemninB, isolated from the Carib- microorganismsproved to possess bioactive bean tunicate Tridedemnumsolidum. 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 interrelationshipsbetween macro- immunosuppressiveactivity. Didemnin is in organisms, such as the sponge, and the mi- clinicaltesting (Klausner 1986). A n o t h e r croorganismsthat colonizethem. secondary metabolitehaving promising prop- Anotherproblem 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 Universityof Califor- protection such as a shell, use chemicals to nia at Santa Barbara researcher who had been deter attackers. However, the defensive investigatingmarine sponges for more than chemicals are not always produced by the thirteen years. Manoalidewas isolated from organism wielding them but instead are the South Pacific sponge Luffariella varia- derived from exogenoussources. For exam- bilis, and belongs to a group of chemicals ple, nudibranchscommonly feed on inverte- called terpenes, although its structure is brates. Somenudibranchs appear to sequester highly unusual (Klausner 1986). Detailed from their prey substancestoxic 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 analgesicproperties, as well as evidenc- on. The nudibranch "Spanishdancer' depos- ing some anti-leukemicand anti-fungalprop- its its eggs in the open, on rocks and corals. erties (Austin 1989). In view of these find- Becausethey containthe bioactivesubstances 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-leukemicand anti-fungalproperties. rently in clinical trials for use against skin disorders, includingpsoriasis.Further investi- MARINE TOXINS. Some secondarymetabo- gation of the same sponge has led to the lites do not display general cytotoxicitybut discoveryof 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, saxitoxinsderived from dinoflagel- anti-inflammatoryproperties. 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 environmentoften 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 Marlnebiotechnology 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 frequentthan shell fish them in medicineor industry. One can imag- poisoning is ciguatera fish poisoning. The ine, for example, that palytoxin, which is a sequenceof events that take place before man nonprotein polypeptidehaving 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, incidentallyingesting the clonal antibodiesprogrammed to attachthem- 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 appropriatelydispensed 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 produceshalitoxin, which inhibitsthe 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 medicinesbecause of side-effects genetics. This dual quality confers on the or other problems neverthelesshave impor- [toxins] an important role in the expanding tant uses as models for the designand synthe- bridge between chemistryand modern molec- sis of other drugs (Colwell 1986). Toxins are ular genetics."(Oliveraand 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 smoothmuscle 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 elucidatethe nerve excitationmech- these organisms do not make the toxin; it is anism. Lophotoxin, which comes from the manufacturedby a marine bacteria that lives gorgonia Lophogorgia, inhibits nerve-stimu- in symbiosiswith Palythoa. Intensiveinvesti- lated contractionof muscle (Colwell 1986).A gation is proceedingto clarify how these bac- bacteriumliving commensallyin 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 neosurugatoxinand prosurugatoxin, 1982). which are powerful ganglion-blockingagents (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 MARINEBIOTECHNOLOGYAND DEVELOPING COUNTRIES

But in nature, enzymes are chemicalsvital to of high heat, tremendous pressure, no light life, catalyzingmetabolic 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, investigatingthe and enzymes are able to function at these relationship between brown alga and an temperatures, which kill most organisms. amoebaof the Trichospaheriwngenus. 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 chlorinatedand bro- enzymecalled Vent DNA polymerase,which minated compounds, she reasoned that has been purified from Thermococcuslitora- through an evolutionary process, certain lis, a type of archaebacterium.This enzyme amoebahad 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 polyethylenesand polyvinyls, are 1000 C (hermostable 1990). Soon it should typically chlorinated and brominated. Dr. be possibleto 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 utilizingconventional indicated that the amoeba would indeed enzymes, tend to be inactivated by heat degrade plastic, she initiated research to higher than about40° C. Since enzymesfrom improve the amoeba's degrading ability. extremophileswill have greater stability and Using mutationby ultravioletlight and selec- last longer at high temperatures than those tion, Polne-Fuller eventually isolated a mu- now used by industry,their availabilitywould tant amoebastrain that has a powerful plastic open up new possibilitiesfor efficient manu- degrading ability. More testing indicatedthat facturing(Tucker 1985). the mutant amoeba would destroy plastic in the field as well as in the laboratory. At this SECONDARYMETABOLITES AS INSECTI- point the university applied to patent the CIDESAND HERBICIDES.Screening of terres- mutant amoeba and contacted industry about trial microorganismsand 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 fundingresearch to clarify the metabolic vary widely; they may block cellular respira- pathwaysof the amoebaby using C,-labelled tion, inhibit protein synthesis, interfere with polymers; these polymersare not availableso 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. Undoubtedlya large variety Rogerson and Gibor 1991). of marineorganisms produce secondarymeta- An exceedinglypromising area of marine bolitesthat have similarproperties, but so far biotechnologyis the studyof extremophiles- littlehas been done in this area. For example, organismsthat 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 compoundcontain- 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 stimulatinggenes' codingfor enzymesthat chemists elucidatedthe toxin's structure and digest fungal cell walls. named it nereistoxin.A synthesizedderivative Chitin, which is the major componentof called cartap hydrochlorideis 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-bloodedanimals 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- Screeningof terrestrial organismshas also sor of chitin and has the largest supplyof 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 applicationsfor it have been developed. and phytotoxins. Some secondary metabo- lites, such as homoalanosine produced by Both research institutes and private com- Streptomycesgalilaeus, have both insecticidal panies undertake or sponsor wide-ranging and herbicidal properties (Vining 1991). As programs for screeningmarine organismsfor with the case of insecticides, we can also bioactive secondary metabolites. Screening expect to find marine organismsthat produce assays may utilize tumor cell lines, DNA and metabolites which act on plants in differing RNA viruses, pathogenicbacteria, or a com- ways, includingherbicidal effects. But so far binationof 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 illustrativeexample 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 chemicalstructure. It is poten- of Marine Sciencesduring 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 fungicidaland pesticidalproper- project's objective is for the contractor to ties useful to terrestrial agriculture.In experi- collect 1000species of invertebrateorganisms ments, chitosan stopped the growth of fungi and mollusksper year and submit the speci- pathogenicto plants, made pea tissueresistant 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-rottingdisease gram remains after sea water is extracted (Hadwiger 1988). from the mass of the collectedorganism. 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 BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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. Thosethat indicatesignificant 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-AIDSactivity using a human lympho- strophic failure of equipment (Costertonand blastic cell line infected with the live AIDS Lappin-Scott 1989). Bacterial corrosion is virus. Preliminaryresults 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 accordingto a NCI spokesman,a number of six months. Piers, off-shore drilling struc- extracts are cytotoxic.The isolation,purifica- tures, and OceanThermal Energy Conversion tion and identificationof the pure secondary devices are especially prone to damage by metabolite from promising extracts are ex- organisms constitutingthe biofilm. pected to take an additionalfive to ten years; Presently there are two methods for pre- then pre-clinical research can begin. venting marine organisms from forming a Some pharmaceuticalcompanies 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 disadvantagesof this usually are more extensivethan those of the method are that these coatings are expensive NCI, includingin additionto screensfor anti- in themselves, they are costly to apply, and tumor and anti-viral activities, tests for anti- as they age they crack, exposingnew 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 consistsin part of a heavy metal such as copper. The metal slowlydissolves in When a manmade object is submergedin 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-foulingpaints are that they pollute grow on a surface rather than in the sur- the marine environment, they are fairly rounding water. If the settlementprocess is expensive, and they pose hazards to the allowed to proceed, the initial film consisting workerswho manufactureand apply them. Of mostly of microorganismssoon attracts mi- course, once encrustationhas 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 damagingoperation. will damage the object via two types of Marine biotechnology research is being actions. First, as an ever increasingnumber directed at solving the problems related to of organisms become enmeshed in the biofilmsand biofoulingand, 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, Researchat AgouronInstitute, California, for biofoulingwill increase vessel drag, thereby example, is aimed at clarifyingthe process of reducing its speed and increasing fuel con- bacterialcolonization, particularly identifying sumption. It has been estimated that a 200 the genes that control attachment. Research micron thick encrustationon 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 discontinueddue the Danish Marine Biotechnology Center. to marketing problems. However, Enzone There various terpenoid compoundsor 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 experimentalcoatings. Some, it is product. The results have been excellent so claimed, are as effective as tin oxide in far, with the adhesive being biocompatible preventingfouling (Cooksey 1991). Yet other and nontoxic (McGuire 1992). scientists, at the Universityof Maryland, are A secondcompany, BioPolymers of Mary- concentrating on the second step in land, took the more traditional approach of biofouling-when invertebrates respond to collectingthousands of the blue mussel, then chemicals exuded by colonizingmicroorgan- extracting the adhesive from a gland in the isms by settling on top of them (Curtin mussel's foot. Approximately30,000 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 chemistsstudied the glue's structure and were organismsto identifyfavorable settlingsites. able to generate sufficientinformation about Research to solve the nettlesomeproblem it to computer-designbiosynthetic glues. An of biofoulingcan, at the same time, be useful endlessnumber of variationsof the basic glue in developingproducts 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 customersfor laboratoryadhesives. However, itself to hard surfaces. The advantageof this BioPolymer'sprocess 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 additionto the two U.S. firms, Univer- Genex Corporationin Marylandbegan inves- sal BiologicalsLimited (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 sufficientinformation about it to enablethem 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 introducedinto the genome immunoassaysand microinjection (Cell Tak of an industrial yeast; then the engineered 1990). yeast was grown in fermenters where it Although marine biotechnology research produced comparativelyspeaking large quan- directed at biofilms and biofoulinghas 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 BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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 compostedanimal wastes biological activity whereby an organism and crop residues since time immemorialnot irreversiblyattaches 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, includingmethane gas and natural the surface (bioadhesives);and (4) the bio- fertilizer. However, attempts to deliberately chemical and biophysiologicalproperties of employ endogenousmicroorganisms 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 adhesivesproduced by the tello 1991). hundreds or thousandsof mollusksother than Bioremediation, and its efficient action, the blue mussel. While most of this research depends on a number of conditionsincluding would be basic research, the lesson from the the characteristicsof microorganismspresent work of Genex and BioPolymeris that one at the site of the process, the chemicalmake- does not need completeinformation about the up of the substanceor substancesundergoing bioadhesive's chemical structure or the mus- bioremediation, the availability of nutrients sel's genetic control over the bioadhesive's vital to the bioremediatingmicroorganisms, productionto develop productsthat are useful the ambient temperature of the environment to transport and specialtychemical industries. wherebioremediation 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 ponentsof the starting material. Microorgan- is released by natural or human action into isms bioremediateby feeding directly on the the terrestrial or aquatic environment will organic pollutant, by breaking down the eventuallybe attackedby microorganisms.To pollutant while they catabolize a primary illustrate, an environmentalmicrobiologist 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). microorganismsand 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 microalgaeand 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 combinebacteria 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 populatedby 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 immensenumbers of microorganismsthat for the bioremediationprocess. The substrate have over thousandsof years evolvedso they is usually quite complex, containing varied are able to utilize petroleum as their sole or chemicalstructures and constituentelements. main source of carbon. Although the time- For example, hydrocarbons may have short span is much shorter, similar situationsexist or long structures; they may consist of a in the soils surrounding long-standingman- 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 developmentand propagation. of carbon and hydrogen atoms and that have More often than not, bioremediation simple structures are usually relatively non- reactions are mixed-culturereactions; that is, toxic and easily decomposed by microor- they involve the simultaneous actions of ganisms. Moleculesthat are highly branched several types of microorganisms.The constit- are difficult to decompose by microbial uents of mixed-culturereactions vary widely, action; those that containmetals, chlorineand dependent on the location where it is taking bromine are toxic to animals, plants and place, the makeup of the reaction mixture, microorganismsalike. 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 extremelydifficult to study. Currently the bioremedial capabilities, the problem of best way to study and research them is matchingorganism 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 particularmicro- uuTrie the sustrate will cosm may be designedto duplicate as closely uayrovidthe major steanc for hes as poil-spiblsite,saypossilDethe eniroinenenvironmentthe and d cdiiaounsconditions oofr continuedirmdaigmcrb,tahealth will depend on the availabil-irbs an oil-spillsite, say the Prince WilliamSound 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 samplesfrom 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 overabundanceof that of the Sound and it would go through an substratesuddenly becomes available,but the identicaldiurnal light cycle. Once the micro- amountsof nutrientsremains constant.There- cosm has been set up, experiments can be fore, large-scale bioremediation cannot be performed on it to clarify what happens if successfulunless additional nutrients vital to one or another physicalparameter is changed; the massivepropagation of the bioremediating if a new microorganismis 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 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

mulated to fit the particular conditions of industrywill includeboth aerobic and anaero- each bioremediationproject. bic phases. For instance, aerobic organisms will be used on the surface, while anaerobic TEMPERATURE. Bioremediationis a com- organisms may be employed to treat petro- plex process involvingchemical and fermen- leum constituents that have sunk and are tation reactions. Chemical reactionsare often amassingon the sea bed. temperaturedependent 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 bioremediationtends to work better in warm- a confinedspace, such as in a test tube, flask, er than colder climates. However, fermenta- or fermenter. An open system reaction is an tion reactionsdo 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 fermentationreactions drive fermentation,are the end productsof an that take place in closed systemsare easier to evolutionaryprocess that has been affectedby perform, study and control. It is therefore temperature. Thus microorganismsfunction understandable that most attempts to use best within the temperature range in which bioremediation for pollution control have they evolved. A petroleum-utilizingmicrobe 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, bioremediationwill usually proceed to bioremediationof 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 chemicalprocesses. crobes that have been propagatedin 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 collectedfrom local sites affectedby the may be used to decontaminate,for example, same or similarpolluting 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 collectedfrom other sites may be used treat municipalwastes. The first would neces- to augment efforts. Alternatively, microbes sitatethe use of aerobicmicroorganisms-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 metabolicreactions via classicalbreeding and selectionprograms where oxygen is of primary importance. The may be employed. main catabolicproducts 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 involvingboth aerobic and anaerobic micro- local biota can expand in numberssufficiently organisms.Anaerobic organismsderive 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 attemptsat bioremediationby 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, bioremediationmay be done waste water treatments.It is significantlyless by microbes specifically designed for that costly than incineration.In general, bioreme- purpose. Scientists at several institutes are diationrequires 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, bioremediationis more environmen- pathways are being altered to improve the tally benign than alternative waste water efficiency of pollutant degradation. Second, techniquesor techniquesused to treat pollut- organismsthat already are efficient in break- ed or degraded soil. Bioremediationis 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 alternativeto bioremediation, fact, the first microorganismgranted a U.S. exceptingnatural processes, because conven- patent (in 1981) was one developed by Dr. tional cleanuptechniques, consisting of skim- A. Chakrabartyto simultaneouslybreak down ming oil from the water surface, moppingup several petroleumfractions. The use of genet- oil by hand using paper towels, cleaning sand ically engineeredorganisms 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, bioremediationposes difficulties treat oil spills. Two types of agents have been while conferring advantages. In regard to tested in the field-biological dispersantsand difficulties, the major limitationof 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 enhancebioremediation because dispersedoil from municipal or industrial sources whose is more susceptibleto attack by microorgan- composition are not completely known and ismsthan is massedoil. On the negativeside, cannot be predicted. An unscrupulous con- dispersants may worsen the effects of the sumer could, for example, chooseto 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, bioremediationof the Surfactants reduce the surface tension of effluenttaking place downstreamwould 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 surfactantproduced by Pseudomo- costly, in terms of both capital expense and nas aeruginosawas found to increasethe rate 46 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

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 resistanceto 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 regeneratingplants 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- terrestrialplant species, so many commercial- hanced oil recovery from oil wells and pollu- ly importantcultivars, such as wheat, barley tion control (Weiner 1985). and oats, have not yet been regenerated. Cell cultureis similarto 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 individualcells. 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 technologiesare 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 speciesare 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 implicationsof does the native plant. Products from terrestri- this capacity is that one plant can be used to al plant cell culture processesrange 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 achievedby 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 consideringalgal cell culture, it is phous mass called callus. A callus can be importantto recognizethe differencebetween further subdivided; the subdivisionscan be this technique and aquaculture. Unlike algal cultured to generate shoots, which in turn can aquaculture,the purpose of which is to grow be maturedin 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. signedand developedto maximizeproduction 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 culturistprecisely 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 Chlorellathat 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 subsequentdownstream process- for the production of pharmaceuticals. Re- ing, consisting of recovery, purification and search done by the company's scientistshas 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 involvingmacro- 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 accomplishmentsinclude modern biotechnologywould not have been the French Associationfor Research in Solar possible without the availability of the poly- Biology developing microalgae for the pro- saccharides from marine macroalgae."(Renn duction of hydrocarbonsand 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 problemslie with inadequatesupplies of agar- accomplishmentby scientists at Amoco Re- producing algae and poor quality control of search Center, Indiana, in using genetic the natural product due to seasonalvariations engineeringtechniques to introducethe bacte- and differing production methods. To allevi- rial genes coding for the rare amino acid ate the unsatisfactorysituation, the European octopamineinto the green alga Chlamydomo- Research Coordinating Agency (EUREKA) nas (Tucker 1985). has agreed to underwritea 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, characterizedand then appropriately ($10,000per kilogram),food coloringphyco- geneticallyengineered 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 internationaleffort, 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 biotechnologyestimates 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 alsouses microalgae versity of Lille and the Instituteof Technolo- as the active ingredient in immobilizedcell gy, AmiensUniversity. Preliminarydevelop- systems, producing via continuous culture ment and pilot plant trials will be conducted hydrogen, acetic acid, dihydroacetone, and in Ireland and Spain (Polysaccharides1991). gluconic acid, all valuable industrial chemi- A few cell culture systems based on cals. In addition, microalgaecan be used to microalgaeare alreadybeing used by industry treat wastewater inexpensively. for the productionof specialtychemicals. For Very little research has been done on example, the Ethyl Corporation in Louisiana marine animal cell culture. As stated in a 48 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

recent review, "Tissues of fishes are amena- complexityfrom temperaturegauges and pH ble to the techniques of modem cell meters to radioimmunoassayand 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."(Hightowerand sor. A typical biosensor consistsof 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-generatingele- 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 conditionsare more exacting for animal cells gauge the reaction by quantifyingthe 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 physiologicalsystems to interest for marinebiotechnology. The first is stress, tolerance of organisms to heat and the chemoreceptor, the heart of which con- cold, cancer biology and environmental sists of biomolecularassemblies involved in toxicology (Rechnitz 1988). In view of its physiological functions, such as smell and possibilitiesfor research and applications, it taste, and in metabolicand 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 investigatorytools 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 substancesranging in chemical substances. complexity from simple salts to pheromones (hormonesthat attract the opposite sex). In Sensors the laboratory, antennules dissected from crabs and connected to potentiometershave Sensors are devices that detect a specific exhibitedinstantaneous quantitative responses substanceor 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 monoclonalor polyclonalantibody means clear to the operator. Sensors also are (see page 7) or a DNA probe is the molecular analytical instruments that react to change, recognitionelement in this biosensor. Modem whether it is a change in temperature, chemi- detectionkits based on one or another mono- cal compositionof reactants or substrate, or clonal or polyclonal antibody have been another parameter. Sensors may have a dual developedto 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 shigellosisand 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. Conventionalsensors commonly employed The second product, the DNA probe, is in laboratories and industry may range in the most exact and definitiveway 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 dinoflagellateswill bloom (prolifer- animalsand 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 dinoflagellatesproduce dead- bacteria, or parasite is determined, then a ly toxins, algae (as well as marine animals complementarysequence 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 demonstratedthe existence ic sequence that gave rise to it. The fact of of some marine bacterialspecies that promote attachment is detected through a procedure the bloom of dinoflagellatesand others that that relies on chemical or radiologicalreac- retard it. The intent is to identifyeach of the tions. A battery of such diagnosticprobes can various retardingmicrobial speciesand deter- be put together in a kit, enablingthe 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, identifyand characterizethe 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 dinoflagellategrowth and propagation. generated in a matter of four to twelve hours. Once this has been done, scientistswill try to 'Te major advantage of biosensors over mass produce this substanceby appropriately conventional sensors are that they are easier genetically engineeredbacteria. 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 dinoflagellatestrains that consti- giving techniciansand engineerstimely warn- tute the red tide. Monoclonalantibodies will ing to adjust reaction rates to achieve maxi- thereupon be constructed for each strain. mum production. As the technologyadvanc- 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 dinoflagellatesbe the point where they will be disposableafter identified quickly. Timely treatmentcan 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 affectedby red tides. For example, medicine. the ChileanUnder-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 JapaneseFisheries 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- poisonedby red tide (Couve 1992).The latter algae aquaculture from red tides (Five-year number is probably an underestimationsince 50 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

many who consumed contaminatedmollusks As was noted in Chapter 1, biological did not report their sicknessor were misdiag- oceanography,while extremely important, is nosed. a field that has been relatively neglected due The marine environmentis highly variable to limitations inherent to present-day detec- and exceedinglycomplex. The inter-relation- tion and measurementtechniques. Biotechnol- ships in the oceansbetween biological activity ogy techniquesthat 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 biologicaloceanography. 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 identificationand quantificationof species, is continuously monitored by sensors em- stocks and populationsof marine organ- placed on anchored and free-floating buoys, isms from microscopicviruses 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 distributionof marine organ- dearth of data on inorganic and organic isms; identification of the mechanisms pollutantsthat 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 manipulationof these on intact microorganismsthat inhabit waters regulatory mechanisms for enhanced on a temporary or permanentbasis. 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 possiblyeven for the bioremedi- and chemical processes in the marine envi- ation of changes in greenhouse gas accu- ronment so they can be adequatelystudied. mulationand the rate and extent of clima- tic change (Initiative1990). Biologicaloceanography (includingpublic health) The report delineates three areas in bio- logical oceanographythat can benefit from Biological oceanography is the study of the applicationof biotechnology.First, bio- "the ecological relationships of organ- technologytechniques can be used to increase isms-their interactions with other organisms our understanding of how the distribution, and with the features of their environment, abundanceand 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 hybridizationand poly- tionships between the environment and its merase chain reaction (PCR) will permit microbial inhabitants is the concern of a scientiststo characterize the genetic makeup scientific field called microbial ecology of populations,to perform comparativeanaly- (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 environmentaltoxicology. knowledgewill 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 commerciallyimportant microplankton (which have recently been fish stocks, such as salmon; developplasmids proven to be responsible for most of the in macroalgaeas 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 particularlyimportant since researchers are impact of microbial activity in the oceans and able to cultureonly 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 employedto rous, oxygen and others that interact with the research how marineorganisms adapt to their carbon cycle; which is a vital determinantof varied environs, clarify interactionsbetween global climatic change and the accumulation species, map the evolution of species, and of greenhousegases in the biosphere. explain the symbiotic relationships between Environmental toxicology has particular organisms. There are many possibilitiesfor 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 clarifyingthe geneticcontrol over the produc- lives, and whose terrestrial and marine envi- tion of enzymesby 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 Massachusettshas 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 milliliterof 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 metamorphosisin shell- "naturally" inhabiting beaches and coastal fish can be identifiedand 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 unquestionedthat huge quanti- opening up possibilitiesfor new products. ties of raw sewageand other waste is dumped Third, biotechnologytechniques 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 interactionsbetween may become discernablerapidly, 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-importantpro- wastes along Chile's 4,200-kilometer long cess wherebyinorganic carbon is taken up by coastlinehas led to many cases among swim- organisms (mostly bacteria and plants) and mers of hepatitis A and E, as well as a few convertedby 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 locatedjust 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. immunofluorescenceprobes 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 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

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 Administrationto market the product. released in the marine environment, the The major applicationof LAL today is in the interactions between bacterial pathogens and testing of injectable pharmaceuticalproducts marine viruses, and the probabilityof 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 numbersof bacterial and viral it does not differentiate between the many pathogens inhabiting waters directly (for types of endotoxinsthat may be secreted by example, while swimming or washing) or bacteria (Novitsky 1984). indirectly (as a result of, for example, eating To concludethis section, two subjectsthat marine animalsthat have been harvestedfrom are related to possible marine biotechnology contaminatedor polluted waters) will suffer applicationsfor 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 biomedicalresearch. 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 establishingcause- the immunoprotectivemechanisms of sharks, effect relationships between pathogens and whichrarely 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 extremelyhigh blood levelof 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- investigatorsperform 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 cationsfrom "classical"marine biotechnology be the source of foodborne diseases due to is related to the detection of toxins important infectiousorganisms or toxins. Seafoodshave to public health. It stems from a 1955discov- been implicatedas the vehiclesfor 10 percent ery that the blood from the horseshoe crab, of all foodbornediseases 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 developedby C. Gram). A specificpathogens and toxins in seafood. Kits purified extract of the blood suitable for based on monoclonalor polyclonalantibodies diagnostic work was prepared in 1964 and seem to hold particular promise in the short was named Limulusamebocyte lysate (LAL). term. As kits are marketedand used routinely After the large pharmaceutical companies in the seafoodindustry, contaminatedbatches showedtheir disinterestin LAL, its developer of seafoodwill be detected early and prevent- started his own companyto commercializeit. 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 Marlnobotochnology 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 researchersare trying to answer this Terrestrialagriculture question. For instance, a team at Canada's Plant BiotechnologyInstitute, Saskatchewan, Cyanobacterand other microalgae, which is trying to improve the cold hardiness of are normallypresent in wet soils, possess the crops subject to damage from frosts that may valuableattributes of nitrogenfixation, 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-phaseapproach in surprise that these aquatic organisms, espe- tackling the project. First, its members col- cially the cyanobacterAzolla, have been used lected the anti-freezeprotein 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 subjectedto freezing. The recently, mass culture technology has been freezingpoint of leaves of treated plants was adapted in India to produce free-livingcyano- 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 meteorologicaldata 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 cyanobacterbiofertilizer use versus chemical -2° C would decrease the number of dam- fertilizer shows conclusivelythe superiority aging frosts from 22 to 5. The third phase, of the first (Metting 1989). While the devel- which is a proposedone, involvesintroducing opment of microalgae as biofertilizer (also the gene coding for the anti-freeze protein called algalization) has so far been done by into an appropriatecrop 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 bacteriumAgrobacterium tumefaciens improvementcould be significantlyadvanced as a vector for introducingthe gene into the through the employmentof advancedbiotech- target plan. However, a key problem remains nology techniques. to be solvedby the Canadiangroup, namely, Comparedto terrestrial life, marineorgan- to clarify the regulatory sequence that con- isms possess characteristicsthat in some cases trols the expressionof the anti-freezegene. are unique, in others merely unusual. For The natural gene is controlledby a sequence example, the world's most salt-tolerantplant 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 inhabitsthe Dead Sea, which contains29 A similar problem may have been solved percent salt (Weiner 1985). The fastest grow- by the U.S. companyDNA Plant Technology ing plant may well be the giant kelp, which Corporation in New Jersey. Company offi- has an estimated maximumgrowth rate of 65 cials claim to have synthesized the flounder centimetersper day (Robinson 1985). And, anti-freezegene 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 transformedyeast's viability after freez- animals, includingother 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 characteristicsinherent produce frozen dough products. In regard to 54 MARINEBIOTECHNOLOGYANDDEVELOPING 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 containingthe gene. It is hoped bigelovii was found to be "...a potentially that fruits and vegetablescontaining the gene valuable new high-yielding oilseed crop for will retain most of their fresh character after these regions, yieldinga vegetable oil high in freezing and thawing (DNAP plants 1991). unsaturatedfatty acids, which is amenable to These findingshave implicationsfor 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 sometimesdevastated 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 temperaturesthan they can now. densely populated,while, concurrently,huge It does not take much imaginationto realize tracts of what are termed marginal lands are that large areas in developingcountries, 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 biotechnologyimmediately 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 developingplants thrive when irrigated by brackish or salt that are salt-tolerantor require salt water for water. Once this developmenthas occurred, life (halophytes). The significant advances the concept of marginal lands will change; that have been achieved in these areas of henceforththe problem will be one of match- research have been reviewed by the NAS ing a land having certain chemicaland physi- (BOSTID1990). More recently, a terrestrial cal characteristics with a crop that is best halophyte was evaluated after six years of suited to grow where these characteristicsare field trials in the Sonora desert, Mexico, present. where conditionsof temperatureextremes and 4 Options in marine biotechnology for developing countries

In view of the limited resources available this fact, we neverthelesshold that countries to most developingcountries, R&D likely to whose bioscientific capability is low could produce applicationsin the short term (oneto 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-termprojects. 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 capabilityin the coun- pinnings of marinebiotechnology to its more try in biology, bioscienceand biotechnology, applied aspects, many developing countries as well as in applied areas, such as aquacul- possessingor controlling marine tracts have ture and fisheries; and (2) the potential of established significant marine-related indus- prospectiveR&D resulting in applicationsfor 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 consistingof 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 AppendixA, research operations may include a research unit, institutes in the fields of marine biology and staffedby a few technicians,who are respon- marine biotechnology-relatedareas are scat- sible for solving immediateproblems pertain- tered throughout the developing world. Un- ing to water qualitycontrol, diseasesaffecting doubtedly, their capabilitiesvary widely; for cultured animals and plants, feed and nutri- example, many have the capabilityto perform tion, and waste management. rudimentary experiments in marine biology, For countriesthat 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 instituteswould technology,two optionsexist. 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 capabilityto perform simpleexper- hiring doctoral-level natural scientists and imentsin 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&Dinvolving transgenic organ- ment may stimulate such development by isms is on an order of one or two magnitudes granting appropriatesubsidies, 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 -

, / ~~~~~~~~~MARINEBACTERIA 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 OptionsIn marlneblotechnology for developingcountries 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 institutionhelps water pearls (McElroy 1990) and fresh water industry by performing problem-solving pearls (Pearls 1992). The expansionof aqua- research or research aimed at enhancing culture, however, will take place through the production. Implementingeither option, or a employmentof mostly well-knowntraditional 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 institutesalready 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 initiativesmight be op triploid and transgenic fish or shellfish to bridge the gap between the research estab- specificallyfor aquaculture, for the purpose lishment and the applied sector by fostering of increasingyields 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 institutionsin developingcountries 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 maturationprocess, 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 possibilitiesfor Asian and Pacific coun- hesion, bioremediation,cell culture, biosens- tries to culture mollusks and export them to ors, public health, and terrestrial agriculture). receptivemarkets 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 culturedlocally. 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 percentof the triploids Countries in Asia and Latin America have were harvested and sold at a premium price. seized on these opportunitiesand 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 findingswould 58 MARINE5IOTECHNOLOGYANDDEVELOPING COUNTRIES

have implicationsfor many countries in the standardsthat 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 mollusksin the SouthPacific, it would proba- prove difficult to procure. But a proposed bly be advantageousto develop an indigenous transplantationof the transgenicmacro-algal R&D capabilityto produce triploids, thereby species to open waters would be likely to foregoingthe need to importthem. raise difficult issues, as discussed on pages Althoughtransgenic fish have been devel- 18. Environmentalimpact assessmentswould oped with favorable growth characteristics, have to convincingly demonstrate that the they are not likely to be cultured on a large transgenicalgal 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 environmentif acciden- adequateregulatory 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 shouldculture transgenic macro-algae in open animals raised in the laboratory. But before systems.The propagationof transgenicmac- field testing can commence, the environmen- ro-algae in a closed system would probably tal impactof the possible releaseof the trans- not be economicallyfeasible. (The culture of genic fish has to be assessed. The results micro-algaeis discussedbelow, in the section from the assessment would be the basis for on cell culture.) regulations governing field testing, which would have to be formulatedand implement- Marine animal health ed by the appropriate national executive and legislativebodies. The undertakingof appro- Fish, crustacean and mollusc aquaculture priate safety testingof transgenicanimals and is important to many developing countries, the developmentof 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-twofinfish, sixty-three be expected in the short or mediumterm. mollusc, eighteen crustacean, and forty sea- Many types of terrestrial transgenicplants weed species are culture in one or more have been developed and have been, or are countriesof the region (Csavas 1989). Con- being, field tested. Several industrial coun- sidering the damage that infectiousdiseases 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 aquacultureseems worthwhile.Some R&D though as far as we know no transgenic is being expended for this purpose. As noted, macro-algaehas been sufficiently developed several vaccines developed via conventional for field testing, such an advanceis likely not means are availableto protect fish and lobster too far off in time. For instance, a first test from infectiousdiseases, and a recombinant could involve a transgenic macro-algalstrain killed type vaccine against IHN has been that has been developedto overproduceagar. developedand 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 transgenicplants in, for exam- being cultured, which is susceptible to nu- ple, greenhouses,and the less stringentsafety merous diseases, and consideringthe present OptionsIn mardnebiotechnology for developingcountries 59

level of effort, much remains to be done in usage in aquaculturewould 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 Americancountries. To illustrate, worldwide and foot-and-mouthdisease, is now taking aquacultureproduction of shrimp reached an place in several nations. Consideringthat the estimated record 690,100 tons in 1991; the regulations for field testing animal vaccines largest producerswere China (145,000 tons), are in place, permissionto 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 suspiciousabout advancedbiotech- about the immunologicaldefense systems of nology, so public opinion may prevent field most crustacean species. The basic research testing in that country. that could provide this informationis lagging, It is probable that R&D is under way in mostly because these problems have little industrializedcountries on recombinantatten- 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 organismthat has in which scientistsin the developingcountries been attenuatedby researchersremoving 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 applicationsshould antibody response in the host withoutcausing 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 technicalviewpoint, R&D in some fore still an open question. However, the of the more advanceddeveloping countries, major constraintsto molluscaquaculture 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 inedibledue 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 establishedtesting proto- biologicaloceanography, below. cols were followed and local laws were adheredto. If the new vaccinesproved effica- Marine naturalproducts cious, local aquacultureindustry 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 developedthat protect- seas to collect marine animals, plants and ed animals from bacterial diseases, antibiotic microorganismsand screen them for natural 60 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

products useful as specialty chemicals or one compoundends up as a marketableprod- pharmaceuticals. The first phase, that of uct. The demands of this development pro- collection, may be relatively easily done if cess are particularlyhigh if the substance of surface and sub-surfaceorganisms are to be interest is intendedfor human use. For exam- collected, or if the target organisms live in ple, as of this writing, DidemninB (see page accessiblesites, such as mudflats or shallow 36) is still undergoing clinical trials twelve seabeds.This type of collectioncan be under- years after discovery. If its developers suc- taken by most countries. However, some cessfullyconclude initial clinical trials, anoth- collectionwill be costly because it is difficult er five years or so might pass before they to perform, for example, the collecting and completeadvanced clinical trials and the drug screening of extremophiles. Since very few is licensed.The entire research, development countries are by themselves equipped to and testingprocess could cost $150 million or undertake the collectingof organisms in the more, which must be paid up front before the abyssalenvirons, this type of activity encour- drug is marketed. The process to develop a ages joint, cooperativeefforts 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, investigatorstend to design screening products area. Almost all countries possess programs that cover a class or category of the capabilityto collect organismsfrom easily activities;that is, antibiotic,antiviral, pestici- accessiblesites; 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 institutionsto do more than rudimenta- cancer and anti-viral substances. In view of ry investigations,its researchers and laborato- the difficultiesattendant to screening, projects ries may consider entering into mutually that seek to collect and screen animals,plants beneficialcooperative agreements with coun- or microorganism should be cooperative terparts in industrializedcountries. Interna- projects, involvingboth public institutes and tional cooperationis particularlyimportant in private companies that already have experi- cases when a substance shows promising ence in this type of endeavor. It bears men- activityagainst dread diseases, such as cancer tioning here that the screening for natural and AIDS, or illnesses affecting the cardio- productshaving interestingproperties is done vascular system. In these cases risks may be in the same manner whether the organism high (in terms of the substancenot fulfilling under investigationis of marine or terrestrial its initial promise), but if testing is success- origin. ful, the payoff probably will be bountiful. Screeningis, of course, only the first step However it is done, naturalproducts develop- of a long developmentprocess. The experi- ment is a long term effort. ence of pharmaceuticalcompanies indicates There are two possible exceptionsto the that out of 10,000 screenings only 20-30 foregoing. The first relatesto toxins. Purified compounds will be identified as possessing marine toxins costs thousands of dollars per interestingbioactivity. These compoundsare 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 OptionsIn marineblotechnology for developingcountries 61

tracts in the tropical seas should consider producing 1 kilogramof sponge comes out to making the investmentnecessary 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 commercialproduct, 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 developingworld; sometimes it scale culturing of the sponge, would harvest is a significantlocal pollutant. Its derivative and clean it, then process it to isolate and chitosan is therefore an inexpensiveresource recover the desired natural product. In addi- that, as we have seen, has possible applica- tion, such an operationwould be enviromnen- tions in agriculture. For these reasons, re- tally benign because sponge aquaculture is search aimed at clarifying chitosan's pestici- essentiallynonpolluting and the availabilityof dal properties should be encouraged as it culturedsponges 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- Bioflimsand blofouling ed, polluting chemical pesticides. However, large investments in chitin processing and The study of the biofoulingprocess, and chitosan production shouldnot be made until the resultingbiofilm, consistsmostly of basic the market demand for these substanceshas research to explain the settling phenomena been determined. and the molecular biology and biochemistry It bears mentioningthat there undoubtedly of organism-surface interfacing. Research are possibilitiesfor 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 eliminatingmarine 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 consideringthe scarcity of data in this area, now have limited uses, mainly for pottery biofoulinginvestigations are not likely to lead making, painting, polishing and washing. to applicationsin the short and mediumterm. 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 informationexists about mussel sources for interesting natural products, adhesives, including the identification of including some that have been developedto some genes that control for their production the point where they are undergoing clinical and the elucidationof parts of the chemical testing. structureof the adhesiveitself. Researchersin In general, sponges are relatively easy to some developing country institutionsshould cultivate, reaching commercialsize in five to be able to build on the already existingscien- 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 experienceof 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 mediumterm. We note, how- the unit consistsof 1188 sponges. The cost of ever, that if the intended application is for 62 MARINE BIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

human use, for applications in surgery or activity, geneticallyengineered organisms are dentistry for example, the same consider- not likely to be used for bioremediationin the ations regarding safety and clinicaltrials that open marine environmentfor the foreseeable were discussedunder pharmaceuticalswould 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 developingcountries capabili- naturally occurring microorganisms, have ties exist that would allowtheir researchersto been developed and their methodologyhas 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 anaerobicfermen- researchershaving employedterrestrial 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 significantreservoir of appropriately trained scientists are available, knowleJge and know-howhaving 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 technologiesfor indigenoususe. 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 pollutioncould commencein 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, investigateand evaluate new groups that most laboratories will be able to enter of pollutant-destroyingmicroorganisms could into algae research for species improvement, also commencein 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 technicallycapable of geneticallyengineering does its work, its impact is likely to be large pollutant-degrading microorganisms for since algal biotechnology is in its infancy, higher efficiencyor a broader range of activi- leaving much room for adding knowledgeto 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 madrneblotechnology for developingcountrdes 63

ular geneticsof some plants, especiallytobac- 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, someknowledge 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 needsto be done on leaved plants (dicotyledons), which include marine plants to clarify their growth charac- the tobaccoplant, led to early success; scien- teristics; explain the molecular and genetic tists developedtechniques that allow them to basis for regeneration; develop vectors to regeneratemost 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 culturetechniques 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 investigatingmacroalgae is utilizestissue 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 mediumto 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 structuresthan 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 industrialistshave gained a vast amount of been developed for inserting foreign genes empirical knowledge about the large-scale into their genomes. Severalplant species have propagation and processing of microalgae. been transformed by inserting functional Scientistsin developingcountries 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 indigenoussites, 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 immenseamount of basic research has been done on terrestrial plants, whichhas laid Biosensors a basis for developmentsleading to applica- tions. To a great extent, this situationreflects As biosensor techniquesare integralto the the amount of effort that has been devoted to performanceof advancedresearch in biotech- terrestrial plant R&D, which is about 100 to nology, many research teams in developing 200 times that given to marineplants. It is no countries are able to construct monoclonal/ 64 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

polyclonalantibodies and manufactureDNA tries, if any, have the science-basedindustry probes. In the laboratoryresearchers employ necessary for designing, packaging and mar- themto detect and identifypeptides, sequenc- keting detection kits based on monoclonal/ es of DNA, and other moleculesof interest. polyclonalantibodies or DNA probes (seethe 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 technologyrequired to produce detector kits. on reagents and operators; and confounding The development of chemoreceptor and factors may cause false positives or, con- immunologicalbiosensors is in its infancy. versely, concealtrue positives. The ability to Some systems work well in the laboratory detect and track a sought-after chemical or performing specializedtasks. But extending organism in the field is largely dependenton their use past the laboratory stage requires operators having sensors/detectorsthat 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 businessventure; 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/polyclonalantibody, DNA probe or biosensor will availablefor general use in the another material, by itself is insufficient. short or medium term. Instead a detection kit is required, consisting of the biosensor, utensilsto hold the sample, Biologicaloceanography reagents to allow the detector and sample to includingpublic health react, reaction vessels or tubes, a read-out device, and positive and negative controls. One consequenceof the public's concern All reagents and implementsmust be pack- about environmental degradation is that aged correctly and adequatelyprotected 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 industrializedcountries. 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 expansivegrowth in developingcountries as realizationin the short term of biosensorsfor it has in the industrializedworld. In some detecting pathogens or chemicals in marine cases, governments in developing countries environs. However, these biosensors would will indeed recognize the importanceof this function efficaciouslyonly when operatedby 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 nmarinebiotechnology for developing countries 65

would receive the training to implementwork freezing. This property, which is coded for programs in this area. The research projects by a single gene, may be successfullytrans- undertakenby these newlytrained 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 conceptuallyattractive projects involvingthe meantimescientists from developingcountries transfer of genes coding for useful properties trained in biological oceanographywould 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, conferencesand 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 oceanographyand 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 needsto be done environmentwill remain largely obscure for on marine plants before it will be possibleto the foreseeable future. apply their important traits or characteristics on land. In particular, scienceneeds to clarify Marine biotechnology applications the genetic control over metabolism in marine in terrestrial agriculture plants, including the production of secondary metabolitesand tolerance to stresses, includ- Certaincharacteristics 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 seemsto protecttomatoes from 5 Building capability in marine biotechnology

A country's scientific establishmentmust termed basic research; it is usually character- have a capability in biotechnologygenerally ized as being performed without having before it can enter more specialized fields, specific applicationsin 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, includingArgentina, 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 somethingthat 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 enhancepresent R&D capabilitiesor develop pilot plant, then in full-scale industrial pro- new capabilitiesin the biologicalsciences; (2) cesses. set up mechanismsfor effectivelytransferring The general requirements for capability- research findingsto the agricultural,industri- buildingin biotechnologyresearch have been al or health sectors; (3) encourage indige- set forth in two reports (McConnell and nous, existing industryto develop the capaci- others 1986; NRC 1990); the more specific ty for applying research; and (4) promote requirements for marine biotechnology re- new biotechnology-basedindustry. Successful search are discussed in two UNIDO publica- interlinkingof these four sets of activitiescan tions (Colwell 1986; Singleton and Kramer be achieved only if the home government 1991). An underlying assumptionof capabili- gives high priority to capabilitybuilding, then ty building in biotechnologyis that the stan- takes the lead by formulating appropriate dards by which excellence in education and policies, committingthe necessary resources scientificendeavors are measured and judged over the long term to implementthem and, if do not change from one place to another. progress stalls, encouraging scientists and Circumstancefluctuate, priorities may differ, scientific administrators to continue their methods for reachingobjectives vary, but the efforts by bestowingappropriate rewards. fundamentalcriteria of excellencein research and developmentdo not change. Therefore, Building R&D capability the same factors apply in capabilitybuilding, whether it occurs in an industrialor a devel- The process whereby a concept (or idea) oping country (McConnelland others 1986). is transformed into a finished product has This being so, capability building in R&D been called the concept developmentprocess anywhere depends, in the first instance, on (Zilinskas 1989). Capabilitybuilding encom- well-educated,well-trained scientificperson- 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 manufactureof products. demanding. Experience shows that it takes Research may be done to advance knowledge approximatelytwenty-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 biotechnologyresearcher, Bufldingcapability 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 conditionsinclude: the natural sciences or engineering. In view of the interdisciplinarycharacter of biotech- * Adequate facilities housing laboratories nology, studentsneed a broad backgroundin and associatedstructures, includingaquar- the natural and physical sciences, as well as ia. mathematics. Thus study should include * Equipmentof sufficientquality and quanti- general biology, cell biology, biochemistry, ty to do the research scientistshave been microbiologyand genetics; the future marine trained to perform. biotechnologistwould find certain electives * The ready availabilityof commonand rare useful, includingaquaculture science, botany, chemicals needed to carry out experi- phycology and zoology. The chemistry cur- ments. riculum should include general chemistry, * A dependable infrastructurefor 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 maintainedequipment equations. Chemical engineeringcourses are repair and maintenanceshops. essentialfor the engineeringstudents; knowl- * Libraries containing a certain minimum edge in the computer sciences is essentialfor number of current books and journals. all students.In graduatetraining students plan * Computer-facilitatedaccess to internation- and execute an independentresearch project al databases and computermail (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 interactionswith 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 Scientistsshould do post-doctoralwork in a abilities. laboratory different from her or his home * Possibilitiesfor professionaladvancement institutionto gain new skills and perspectives. based on ability and quality of work. Similarly, engineers should obtain practical * Availabilityof sufficienttime and funds so experience at up-to-date pharmaceutical, that the scientist can communicate elec- chemical, or genetic engineeringcompanies. tronically with colleagues in the interna- Once scientists have completed their tional scientific communityand 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 unnecessaryadministrative or quite complex. Essentially, those who study bureaucraticprocedures burdens. long and hard to become scientistsrightfully * An equitablepeer 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- * Possibilityfor an individualscientist 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- * Supportby well-trained,highly motivated 68 MARINEBIOTECHNOLOGY AND DEVELONG COUNTRIES

technicaland secretarial support staff. or three different disciplines. A team * An equitable teaching load; that is, one undertaking research in medical biotech- that allows sufficienttime for research. nology would consist of five to ten spe- * Individual incentives, such as small cialists in biochemistry, cell culture, grants. genetics, monoclonal/polyclonalantibody construction,microbiology, and molecular A significantresearch effort in biotechnol- biology.Depending on personalpreferenc- ogy is best undertakenby 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 expectedto carry out The principle behind the idea of "critical basic research besides applied research. mass' is that five biotechnologistswork- Scientistsat university laboratories may, ing in the same place are more effective for instance,be expectedto perform basic and productivethan the same five biotech- research, or research aimed at solving nologistsworking in five different places. fundamental problems or adding to the This is related to the benefit derived from store of fundamental knowledge. Some- frequent interactionand 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 techniquesto solve a certain it will take time and effort, which mustbe problem, it is essentialthat a team consist consideredwhen determiningthe size and of biotechnologistsfrom a range of disci- compositionof the research team. plines. For example, a successfulteam of workers in biotechnologymust have at its * Whether the team will have a training disposal several independentscientists in function. Obviously, if a scientist has each of the following basic disciplines: teachingand training duties, less time will biochemistry,microbiology, immunology, be availablefor research. The more inex- cell biology, and biochemicalengineering. perienced the student, the greater the Dependingon 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 specializedfields such as genetics, virolo- is long, say two years, then the trainee gy, pharmacology, plant science, animal can be expectedto contributeand 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 considerationswhen technicianscould take on one or two long- attemptingto 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 discussionof them here (a summary is pro- with five Ph.D.-level scientistsfrom two vided in AppendixB). Bulding capaebty In madne biotechnology 69

Applying research present informationabout potentiallymarket- able products to those who may wish to Somewherealong the conceptdevelopment 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 introducingnew or improved biotechnology systemic barriers that prevent technology techniquesinto their processes shouldmake 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 counterpartsto 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- overcomebarriers; somewhatlike two crews search being performed in public laboratories constructinga 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 indigenoustechnology produc- the advanceddevelopment 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 laboratoryfor tutes. It is imperativethat 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 inventionstheir 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 complishmentswill remain tantalizingprom- 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. Universitiesare changing biotechnology research can theoretically everywhere, even the ancient universitiesof benefit technology end users in established, Europe that once epitomized the academic traditionalfirms 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 chemicalsor extracting natural resource. basic (as contrastedto appliedresearch done However, by far most firms in developing usually by industry). In biotechnologyespe- countries have capabilitiesonly to manufac- cially, findings from so-calledbasic 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- developmentunits, which limits their ability fies the molecular control in a cell that pro- to absorb or adopt results from indigenousor 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 advanceddevelopment lab- interest to industry. Unless the researcher, oratories, as discussed in the previous para- and the university employingthat researcher, graph. is willing to forego a possibly significant The secondtype 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 BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

tries. For example, in Peru there has been Beginningwith the first step in the con- only one biotechnology company, Bio- cept developmentprocess, research at univer- ingenierfa Aplicada. This was also the first sities and institutesin developingcountries is company in Peru to set up a direct link be- supportedalmost entirely by governmentsand tween industry and university; and for a time researchers are mostly government employ- BioingenierfaAplicada was able to exploit for ees. As is well known, public universitiesin profit the indigenousnatural resources of that most developing countries are underfunded country on a sustainableand 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 approximatelythe same education and It is important to encourage the estab- responsibilities).In a time of severe budget- lishment of biotechnologycompanies in the ary constraints it is difficult to correct these developingcountries becausethey 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/technicalbase upon which a sci- tant vehicle for the developmentand commer- ence-based industry, such as biotechnology cialization of biotechnologyproducts in their industry,can grow and flourish. Accordingly, countries. However, as is the case of technol- a significant strengtheningof research can ogy transfer units in industry, governments only come about if governmentstake the hard have the major role in creating the economic decisions to raise new funds or divert scarce climate conduciveto the entrepreneurshipof funds from other programsin order to streng- biotechnology-basedindustry because only then scientific institutionsand finance neces- they can adopt measures that encourage sary research. people to make the risky investmentsrequired Supporting research and researchers is, to establish that industry. however, only the first of several activitiesa governmentmust 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 concepthas 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 reachedwhere funds for the three most important are government the advanceddevelopment or the concept is funding of basic and appliedresearch, scien- required. This point is crucial because devel- tific personnel availability and the level of opmentwill 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 investorswill perceive a venture at tence and adequacy of health, safety and this early stage of development as being a environmentalregulations, the operationof an risky investment. A government is probably equitableintellectual property law, and effec- the only source in most developingcountries tiveuniversity-industryrelations (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- demonstratesthe vital role of government, to ernments must make funds availableto indus- make certain that economic, legal and politi- try, either as grants or low-cost, long-term cal conditionsfavoring capability building are loans, so it can set up advanceddevelopment present in the countrywhere it is being done. laboratoriesand pilot plant operations. Bulding capability In marine biotechnology 71

Tax laws may be designed to encourage formulated by OECD (for a general discus- investmentsin 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 designedto promote investments when we move past research. Most develop- in R&D by allowingthe companythat 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 shouldbe 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 shouldbe health or industrialactivity may have implica- designed so they do not unduly tax profits tions for bioscientificor bio-industrialactivi- from profitableinvestments, thus appearingto ties, but national agencieswill 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 companieswill be unable to adequatelyplan wide applicability, such as the PCR (Korn- for the development,testing and manufacture berg 1991). This is not the place to debate of biotechnologyproducts. If they are unable these issues, it is however appropriateto 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 inventionsunless they are assured technology R&D in most developing coun- of exclusive, long-termrights to those inven- tries. Accordingly, governmentsmust 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 concernbiotechnology inventions. If governments wish to set up Discussion biotechnology-basedindustries, they will find it well worth their while to consult with the While keeping in mind the conceptof the World Intellectual Property Organization in developmentprocess, and its starting point in Geneva on this matter, and to analyze the research, this report emphasizesthe applica- BudapestTreaty on the InternationalRecogn.- tions of marine biotechnology.For one, the tion of the Depositof Microorganismsfor the term "biotechnology" implies application. Purposes of Patent Procedure in terms of its Further, applicationis the goal of most assis- own national interests. tance provided by the World Bank to devel- Usually research is not effectedby lack of oping countries. However, this should in no regulationssince universitiesare often auton- way be interpreted to mean that basic re- omous in developingcountries, enabling them search is unimportant to, or an unnecessary to adopt their own rules governingR&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 typicallyperformed at teaching NIH guidelines or the generic regulations institutions provide training in fundamental 72 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

techniquesto the scientific/technicalperson- ment promotingcontacts 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 biotechnologyindustry and firms that utilizes industries. biotechnology techniques, and give expert Besidesthe factors that directly affect the advice to governments. Second, in the course developmentof options for developingcoun- 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 solutionrequires basic that accrues to developingcountries that have research. Third, basic research is a type of attained significant capabilities in marine "safety valve" to imaginative scientists, biotechnologyis that their level of expertise providingthem with the mode for expressing in marine and oceanographic biology will their creativity that at times engendersunex- simultaneouslyincrease markedly. In particu- pected results important to the applied sec- lar, as capabilitybuilding encompassestrain- tors. Clearly, any countrythat wishes to have ing, more scientists in developing countries an in-depth, encompassing bioscience and become trained in biological oceanography biotechnologyinfrastructure requires a strong and marine biodiversity; individuals who basic research component in the biological thereafter will be well-placedto perform the and biochemical sciences (McConnell and investigationsand technologicalassessments others 1986). that will form the basis for the formulationof Nevertheless, as shown by the historic national policies and regulations to protect example of developmentin Japan and, more and manage indigenous marine resources. recently, by Hong Kong, Singapore and the Further, these experts would be well Republicof Korea, a country that emphasizes equipped to serve as consultants to national appliedR&D and, concurrently,constructive- delegations at international meetings and ly imports and adapts foreign technology is conferenceswhose aim will be to deal with likelyto benefitthrough acceleratedeconomic 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 ingredientsto enter this (Initiative1990), input from scientistsknowl- field are often present or are readily attain- edgeable about marine natural sciences will able: biotechnologyis relativelyaccessible to be required when politiciansformulate 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 universitiesof most countries effect, the destructionof ozone in the strato- have ongoingteaching and researchprograms sphere, the increasing levels of toxic agro- in biology, biochemistry,marine biology and chemicals in ground water and oceans, and the ocean sciences, which providesa 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 establishedaquaculture, 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 possiblethat a country measuresthat 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 nongovernmentalorganizations (NGOs) provided major funding to developingcoun- 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 fundinghas also related areas. Some agenciesprovided much been directed at expanding aquaculture in informationabout their work, others did not developingcountries. 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 productionof food; few includedresearch. Of in marine biotechnology-relatedareas. the thirty projects that did include a research We stress that most of the informationwe component, nine used laboratory facilities, present below was provided by the agencies but only one consistedsolely of research. In themselves; in some cases we were able to general, research is done to improve aquacul- supplementit through interviewswith agency ture systems, to identify new professionalsadministering pertinentprojects. species/aquaculturesystem combinations, and From the responses we received it is clear to identify indigenous species that may be that with the exceptionof 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 supportingor are reachedtheir maximumsustainable yield, and involved in marine biotechnology-related that some are beyondthat stage. Beginningin 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 agenciesare dealt with in alphabetical cal look at the future of aquaculturedevelop- 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 SeafarmingDevelopment and related areas. The second part contains our DemonstrationProject of NACA in Thailand, thoughtson possible ways that the agencycan which aims to overcome problems that beset incorporatemarine biotechnologyin 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/FAQRegional 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, Philippinesduring August 1990. the world's food supply. In line with this The FAO indicatedthat it is not sponsor- 74 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

ing any marinebiotechnology projects. It has, logy's long history in the South East Asian however, been sponsoring some research in countries, projects amalgamatingaquaculture 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 InvestmentBank (EIB), the oped, it could open the way for a FAO pro- European Bankfor Reconstructionand Devel- gram that would include marinebiotechnolo- opment (EBRD), the Andean Development gy. In addition, as is made clear above, Corporation (Corporaci6n Andina de marine biotechnologymay be used to signifi- Fomento), and the African Development cantly increase yields from aquaculture Bank (AFDB) are, or have been, involvedin through the genetic improvementof 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 missionof improvingfood bly assessproposals for marinebiotechnology 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 significantinitiatives in marinebiotechnology IntergovernmentalOceanographic in the near term. (FAO's involvementin 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 secretariatis sited at the headquar- The IDB has done extensivelending 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 concertedactions 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 activitiesthe IOC biotechnology.The IDB would probably be is involved with includesupporting the global interestedin providingloans for activitiesthat sea-level observing system (GLOSS); the would improve current aquaculturepractices joint IOC-WorldMeteorological Organization or set up marine natural product development program called Integrated Global Ocean (Peacock 1991). System (IGOSS); ocean science research on The Asian DevelopmentBank (ADB)has living resources in cooperation with FAO; been investingin aquacultureand fisheriesin research on nonlivingresources with various countrieslike 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 InternationalAtomic Energy projects (see discussion in the section on Agency through the Global Investigationof ICLARM, page 82). In view of biotechno- Pollution in the Marine Environment pro- Ma/or internationalagencies and blotechnology 75

gram; and the global mussel watch (see page consisting of thirteen eminent scientists, 4). An important new initiativeis 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 affiliatedcenters: in Algeria, Argenti- constructprediction 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, Venezuelaand Yugoslavia.Applica- long-term goal is to develop IOC programs tions from Iran and Turkey are pending. that will permit managementof 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 developmentis marinebiotechnology, but the developmentof 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 marinebiotechnological 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 employmentof bio- cally, a joint UNEP/ICGEB program on sensors. There is also a strong desire at IOC biosafety commencedin 1991. Its two initial to coordinateseveral oceanographicactivities activities consisted of courses offered in to enhance our understanding of living and Trieste to researchers from developingcoun- nonliving marine systems. Since marine tries: a three-day course was held July 1991 biotechnologyopens added opportunities to and attended by thirty scientists called "Ge- further understand marine environments, netically Modified Organisms: Safety in the there exist possibilitiesfor cooperating on a Laboratory and the Environment" and a variety of marine research related activities subsequentthree-day course with fifty partici- (Kullenberg 1991). pants called "GeneticallyModified 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 establishingthe ICGEB, which is presented December 16-20, 1991 called now operational in two components, located "Marine Microbiologyand 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 mediumterm the ICGEB is not likely affiliated centers in countries around the to becomedeeply 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 representativesfrom 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 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

addition,as scientistsin developingcountries United Nations Conference on learn the techniques of risk assessment and the Environment and Development risk managementat 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 developmentpro- 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 indicationthat IMO sponsors marine to govermments,IGOs and NGOs, includes biotechnologyor marine biotechnology-relat- the item "EnvironmentallySound Manage- ed projects. ment of Biotechnology". In the course of our research we have UNCED's Preparatory Committee found out that various agenciesare preparing (PrepCom)was establishedin 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. cludingthe atmosphere,forests, land resourc- Apparently, IMO will be one of the agencies es, desertification, biodiversity and biotech- consultedfor such work. Further, IMO has a nology; Working Group 2, chaired by Dr. good knowledgeof the needs of wastedispos- Bukar Shaib from Nigeria, addressedoceans, al facilities and its relation to the shipping freshwaterresources and the managementand industryand is experiencedwith 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, includinglegal and institutionalissues, areas of marine biotechnology: bioreme- human settlements, technology transfer and diation of ship wastes, bioremediationof oil Agenda 21. spills and biofouling. Once involved with Chapter 16 of Agenda 21 identifies five enhancingbioremediation, IMO couldsupport 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 enablingmechanisms for the devel- are very toxic to marine life. If biological opment and environmentallysound applica- films that prevent the propagationof 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 intenselytrafficked waters between biotechnology and biodiversity, could be reduced. covering the forest, soil, freshwater and Majorinternational agencies and blotechnology 77

marine sectors (UNCED 1991). cate an interest in marine biotechnologyand Marine biotechnologyis not mentionedin 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 movementto includethis field while carrying investigating irrigation schemes that utilize out the other Agenda21 biotechnologyobjec- nutrient rich sea water to grow salt water tives. In particular, marine biotechnology resistant plants. This type of project could could help connect biodiversityand biotech- benefit from the application of marine bio- nology. In the marine area, one of the best technologytechniques. ways to do so is through biological oceanog- raphy (see pages 64 and following); that is, United Nations Educational, Scientific to use advanced biotechnologytechniques 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- networkfor the exchangeand preservationof ed environment common to coastal develop- industrialmicroorganisms and to train micro- ing countries and for preventingfuture pollu- biologists. In 1970 UNESCO acted to estab- tion through appropriatetreatment of wastes lish a networkof MicrobialResource Centers from land and water-based sources. (MIRCEN), which was indeed establishedin 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 aquacultureprojects in maintainingcell culturecollections, MIRCEN developingcountries, includingChina, 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 includeR&D whose objective vanced techniques of biotechnology, and in was to geneticallyimprove stocks, for exam- general seeks to improve the qualityof life in ple, the GIFT project at ICLARM (see page the developingcountries 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 approximatelyevery 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 professionalsthat we have interviewedindi- microbiology/biotechnologymay be directed 78 MARINEBIOTECHNOLOGYANDDEVELOPINGCOUNTRIES

for the benefit of developingcountries 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 BiotechnologyAction Council,which admin- West and Central Africa, the East African isters a research and training program in seaboard, the Pacific coast of SouthAmerica, biotechnologyrelated 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 activitiessuch as the ships, and in 1993 hopes to fund five profes- monitoring and control of marine pollution, sorships. as well as the coordinationof pollution pre- In the marine field, UNESCO cooperates ventionstrategies. in the work of the IOC to promote marine As is the case with other agencies, UNEP scientificinvestigations (see above), including is not presentlysupporting marine biotechnol- the Coastal and Marine Program (COMAR); ogy projects. The question then is what help has established Man and the Biosphere marinebiotechnology may offer the agencyto Programme (MAB)to develop a basis for the fulfill its mission. One possibilityof potential rational use and conservation of terrestrial importance are biosensors. The joint and marine resources; and is active in the IOC/UNEPGroup of Experts on the Scientif- InternationalHydrological Programme, which ic Aspects of Marine Pollution (GESAMP) is developinga scientific basis for the man- has been exceedingly active in determining agement of water resources, includingbiore- some of the problems of marine pollution; its mediationof polluted waters. work could be enhanced through advanced While UNESCOhas not been involvedin biotechnologydetection techniques. Another marine biotechnology, without doubt, possibility is for UNEP to investigate and UNESCOcould make an importantcontribu- clarify the possibilityof usingbioremediation tion to capabilitybuilding in marine biotech- to treat pollution in developingcountries and nology R&D since it is the lead agency in the to adapt bioremediationtechniques for the UN system for helping countries improve treatment of waste water from land based their educational institutions and basic re- sources. (UNEP's involvementwith biosafety search institutes, and in view of its wide- is discussedbelow in the UNIDO section.) ranging programs related to biotechnology generally. In addition, UNESCO is well United Nations Industrial placed to identify counterpartinstitutions that Development Organization could cooperate in marine biotechnology research having international significance.It UNIDO's interestin marinebiotechnology also could inform policy makers in coastal is of rather long duration, beginning in the and island developing countries about the early l980s. The agencyapproaches this field possibilities that marine biotechnologyhold from two directions-from its multifaceted for them through UNESCOpublications and biotechnologyprogram and from its interest UNESCO-supportedconferences. in setting up a marine sciences center. UNIDO supports numerous projects in biotechnologythat focus on appliedR&D and United Nations EnvironmentProgramme on pharmaceuticaland industrialapplications. Developmentsthroughout the world in bio- Some of UNEP's most significant activi- technology, includingits marine aspects, are ties are focussed on the marine environment, reported in the quarterly publicationGenetic Majorinternational ogencles and biotechnology 79

Engineering and Biotechnology Monitor, broad agreement among countries where which is sent free on request to scientists in advancedbiotechnology R&D is done on the developing countries. However, UNIDO's level of control appropriatefor research and interest in marine biotechnology became large-scalecontained use of geneticallyengi- manifest when Dr. Rita R. Colwell was neered organisms. Similarly, products pro- commissioned to write the report Marine duced by conventionallydeveloped or geneti- Biotechnologyand the Developing Countries cally engineered microorganisms for con- (1986), which discussesthe rich promisesthe tained use are accommodatedunder existing field holds for developing countries and nationalregulations governing drugs, food or outlines an approach whereby they can build environment.However, no such international the requisite capabilitiesto fulfillthese prom- unanimityexists in regard to the testing in the ises. Dr. Colwell's ideas were expanded on field of geneticallyengineered animals, plants in 1989, when the report "Biotechnologyof or microorganisms.Because of this predica- Marine Algae: Opportunitiesfor 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. commercialfirm in Argenti- written, on the specific uses that marine na, a dispute arose about the adequacy of biotechnology has for the Mediterranean safety precautionsof the test. The motives of countries. the firm conductingthe test were questioned, UNIDO is also setting up a program on the implicationbeing that testing was being bioremediation and oil recovery. Primarily done where local regulations were weak or intendedfor countries belongingto 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 scientistsfor improvingstrains of and products where there are few legal hin- oil degrading microorganismand using them drances. Second, and more commonly, be- for oil spill cleanupor enhancedoil recovery. cause regulatory agencies have been unclear Of relevance to marine biosafety is on how to proceed, they have held up the UNIDO's initiativeto establish an interagen- field testing of potentiallyvaluable 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 recommendedthat the agenciesrepresent- nologyresearch, bioindustryand 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 InternationalCode of Con- two years WHO and FAO joined it. The duct for the Releaseof OrganismsInto 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 mentionedabove, there is Resource for the Release of Organisms 80 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

Into the Environment. UNEP is the lead treatmentof a specifictopic in marine indus- agency for establishing the Resource, trial technology; the second contains capsule which will collect and store all available reviews of topical developmentsin the field informationabout deliberate release activi- (includingmarine biotechnology) and a listing ties throughout the world. Its steering of technical conferences and meetings. Free committeewas set up in March 1991; its subscriptionsto the monitormay be requested first responsibilityis to take an inventory from UNIDO's Industrial TechnologyDevel- of existinginformation sources on release opment Division. activities. While UNIDO has not defined a work program for itself in marinebiotechnology, it Draft and publish a biosafety manual. A provides technical assistance for marine manual for principally the developing biotechnology-relatedprojects and it has countries will be prepared jointly by made certain that biotechnologycomponents 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 compilationof national and international componentsmight 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 influencedby attempts are made to harmonizebiosafety pollutantsfrom ocean andland-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 includeprojects aimed at and its applications. the collectingand 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 focusseson 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 InternationalOcean Institute may marinebiotechnology-related areas, including be set up as early as this year. As now fisheries, aquacultureand generalbiotechnol- planned, a componentof the institute will be ogy. Optionsfor future World Bank activities devoted to investigatingthe industrialpoten- in or related to marine biotechnologyare set tial of marine biotechnologyfor 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 IndustrialTechnology 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 advancedmaterials, biotech- countries,this studywas 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 internatlonaiagences 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 industrialcountry (1989FAO statistics 1992). includedcomponents for aquaculture. One important objective of the study was to In terrestrial biotechnology, the major determinehigh priority researchneeds related support provided by the World Bank is relat- to fisheriesand aquaculture,assess the capac- ed to agriculture R&D performed at the ity of certain developingcountries 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- capabilitiesof many African, Asian and Latin bandry, managementof natural resourcesand American countries, the development of germplasm preservation. The World Bank tropicalaquaculture, international cooperation provided about 15 percent ($32 million) of in fisheries research, and research needs of CGIAR's 1990 budget of $235 million. small-scalefisheries. One finding of the study CGIAR's research purview includescom- 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 biotechnologycomponent. producing useful results in as short time as In general, World Bank activities in the possible. Traditional agricultural techniques aquaculturesector has expandedgreatly in the are at the current stage of development the past decades. These activities might occur safest and most efficient systemsfor increas- either as a sub-componentof larger fisheries ing the productivityof crops (as evidencedby 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 supportedproject has found, the investment banks and donor organizations, biotechnologytechniques more advancedthan 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 competencebetween the World (World Bank 199la). The more sophisticated Bank and some of the regionalbanks, 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 aquacultureprojects. introducedat 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 Yugoslaviafor carp. became a member of the CGIAR network in More recently, the World Bank provided 1992. It was established in 1977, largely 82 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

through a grant from the RockefellerFounda- 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 managementand particular, the study, among other activities, marketingof aquatic resources in developing analyzedthe regulatoryand intellectualprop- countries. ICLARM has four basic pro- erty issues as they relate to biotechnologyand grams-aquaculture, coastal area manage- developingcountries. To sum up the recom- ment, captive fisheries management and mendations, in regard to regulations, the information services. Its beneficiaries are report suggestedthat developingcountries set mainly small-scale fisheries and traditional up institutionalbiosafety committees in scien- aquaculturists. tific research institutionsand establishnation- 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 biotechnologyresearch, testing which is being carried out in cooperationwith 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 investmentsin biotechnology,both by tilapia aquaculture due to a deteriorationof local private sector companies and by trans- tilapia broodstock in the Philippines, caused national corporations. Each country needs to in part by the introductionof 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 favorablecharacteristics tions, and then license (freely, if appropriate) (Guerrero 1991). these inventionsfor 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 1981-1987was Outside of agriculture research, some $575million for twenty-onenational projects. other projects have education, science and We cannot determine exactly how much of technologycomponents that include biotech- this moneygoes 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 AgriculturalResearch, the AustralianInterna- 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- Majorinternational agencies and blotechnology 83

ed at about $600 million, with funding to be with agency personnel, there seems to be provided equallyby the Bank and Brazil. The three major reasons for this situation. First, objectivesof this large project is to strength- the professionalsstaffing agenciesare mostly en the managementof Brazil's science and unfamiliar with biotechnologygenerally and technologysector; strengthenBrazilian 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 technologycould be constructivelyapplied 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 technologyoption to policy makers in devel- PADCT II, aims to strengthen biotechnology oping countries. Second, policy makers in for applicationsin human health, agriculture, developing countries and their technical animal husbandry and industry. While the advisers, while often awareof biotechnology, major portion of these funds will support are uninformed of marine biotechnology,so scientific research and technology develop- they are not in a positionto request assistance ment projects, a significant portion will be in this field. The low level of awarenessof used for humanresource development, mostly marine biotechnology stems from the third specializedtraining in-country and overseas. reason, namely marine biotechnology has Lessons from the PADCT project are ex- receivedlittle publicityso 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 massivelyreported on throughoutthe world, project that is being developedfor Mexico to information about marine biotechnologyhas completely reorganize that country's main not spread much beyond researchers in the agency supporting science and scientific field and a few industrialistsseeking to apply research, the Consejo Nacionalde 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 implicationsgeneral biotechnologyhave Mexican Science, is being worked out as this for developingcountries has been debatedby is written. scientistsand 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 influentialstudy on biotechnology cussed in this chapter provide technicalassis- for development (BOSTID 1982; UNIDO tance in general biotechnologyor in marine 1981). No such pivotal events relevant to biotechnology-relatedareas (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 supportsprojects in marine bio- point for the promotionof marinebiotechnol- technology.From the interviewswe have had ogy in developingcountries. 7 Exploring World Bank options for investments in marine biotechnology

In this chapter various options are ex- endemicdiseases from damagingthe cultured plored for World Bank assistanceto develop- animals or plants. Research may be done to ing countries for sustainabledevelopment in develop diagnostic methods for early detec- marine biotechnology. Three approaches are tion of diseases, for vaccines to prevent the suggestedthat are already being taken by the occurrence of diseases, or for therapeutic Bank in other contexts: scienceand technolo- drugs to treat diseased organisms. The ele- gy lending; support of environmentalobjec- ments common to Bank projects would have tives; and support for private sector develop- implicationsfor this component:it would take ment (PSD). into account local conditionsand would deal with a local problem, it would be an innova- Science and technology lending tive approach to anticipatingproblems 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 technologyand its adaption to fit whereby the Bank supports science and tech- local circumstances.Similar examplescould nology (Weiss 1985). Three of these mecha- be given for future projects involving waste nism has implicationsfor capabilitybuilding water treatment,pollution control and allevia- in marine biotechnology. tion, natural products development, and public health in the coastal zone. Technicalassistance for Many opportunitiesexist for this kind of technologicaldevelopment assistance;we will present one examplehere. 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 technologicaldevelopment. As a rule, Bank imports. In particular, Qatar imports soil officials try to ensure that developmentpro- 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 indigenousalternative 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-basedtanks or ponds the technologicaldevelopment 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 broughtto the attentionof 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- circumstancessuch as these would seem to search component to, for example, prevent make business sense. ExploringWorld Bank optlons 85

Projea lendingfor science and technology niques and approaches. For example, marine biotechnologymay be introduced into aqua- culture, natural resource exploitation and Some loans are aimed at directly enhanc- other marine-relatedprojects being undertak- ing the scientific and technologicalcapability en in Ecuador, Indonesia, Malaysia, Philip- of a country. The Brazilian ScienceResearch 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 biotechnologybe how the Bank may assist biotechnology deployed in such a manner that it fuses with capabilitybuilding in developingcountries. 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 mentionedabove leads to the formu- can take to support science and technology lation of projects for strengtheningcapabili- for developmentis 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 technologytransfer that are similar include biotechnology components to, for to agriculturalextension services that are al- example, perform genetic studies on target ready commonly found in developing coun- organisms, clarify the movementsof pelagic tries. Specifically,public institutes, whether fish, prevent and diagnoseimportant diseases university or national research laboratory, afflictingtarget organisms,or improve repro- should be encouraged to set up technology ductive success among culture animals. transfer units accordingto the terms that are The past history of the World Bank's outlined in Chapter 5. This process can be science and technologylending shows that it enhanced by the World Bank making avail- is primarily directed towards countries with able resourcesto governmentsso they can set mediumincomes. Marine biotechnologyis 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 numbersof 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 biotechnologyrequires a sophisti- gestion, it might find a useful model in the cated research infrastructureand 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 NationalOceanic and AtmosphericAdminis- and technologylending to initiate or enhance tration (Ragotzkie1988). marine biotechnology might be aimed at Sea Grant operates through twenty-nine countriesthat alreadyhave scientificinstitutes coastal programs, involving hundreds of wherein sophisticatedbiotechnology 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 excellentmechanism for universities, the second instance, the Bank could target industryand governmentto pursue coordinat- countries that have lower level capabilitiesin ed efforts in R&D, its major function is to the biological sciences, but in which large transfer marine science and technologyfrom marine-related projects are under way that laboratories to technology end-users. It does may benefit from marine biotechnologytech- so by mobilizing support for practically 86 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

orientedresearch projects, and then acting to potentialprojects in three ways. First, marine make informationabout these projects avail- biotechnology techniques may be used to able to industry. The performanceof 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- Supportto internationalresearch somes and/or genes of marine organisms to see if they have suffereddamage 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 determinethe origin of a salmoncaught 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 situationnow 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 internationallaw. 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 encourageIARCs 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, planktonicforms are difficult to ascer- taken up, it might prove cost effective for tain with present methods. These forms can CGIAR to upgrade existingnational 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 determinationof which Support of environmental objectives reef species are diminishingand 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 planktonblooms. project should be carried out at all, in other The marine biotechnology techniques cases they dictate how projects are imple- related to bioremediationmay, of course, be mented. Marine biotechnology may affect used directly to clean up polluted marine Exploring WorldBank optfons 87

environs, but we consider this possibilityin research sector. To illustrate, say that the the next section. World Bank considers a project to establish Relating directly to environmentalinitia- an automobilemanufacturing plant in a devel- tives, there is at this time much discussion oping country. Althoughmany 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 designatedas high four sectors of the GEF will includeactivities technologies, the implementation of this where marine biotechnologycould be compo- project could be done wholly without the nents. One is international waterways and involvementof universitiesor their research- marine environmentsand the other is conser- ers because the technologiesare known, their vation of biologicaldiversity. On improving implementationrequires engineersand techni- marine environmentswe have already men- cians, and their operation will be the same tioned the reception facilitiesfor wastes that regardless of where it is done. the GEF is committedto constructing.Assist- The situation with biotechnology-based ing cleanupof accidentalspills 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 improvementsof monitoringtechnologies that industrial organisms must be employed for marine biotechnologyhas brought about. applied purposes. This impliesthat the natu- Marine biodiversityis also covered by the ral phenomenaunderlying these processes are GEF. There shouldbe possibilitiesin 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 scientistselucidate a natural show the value that biologicaldiversity in the phenomenon, they simultaneously generate seas have for commercializingsea products the knowledge that can drive an industrial might improve the interest of developing process. For example, when a researcher countries to maintainingtheir living marine locates and characterizesa 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-basedindus- World Bank goal because it fulfillsthe Bank's try cannot be establishedin a country unless primary objectives of reducing poverty and there is a close relationships between the raising standardsof living. The four types of researchestablishment and the appliedsector. activitiesthe 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 complishmentsshould lead to PSD whether it laboratoriesshould be linked with technology is in agriculture, food industry, chemical end-users in industry, agriculture and health. industryor biotechnology-basedindustry. 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 universitiesand nationalresearch institutes that biotechnology-basedindustry, wherever reaching out to technologyend-users, possi- it is to grow, must be closely tied to the bly through a Sea Grant-like approach, has 88 MARINESIOTECHNOLOGYANDDEVELOPING 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 Technologyand tbneirwnu indigenous industry, could make funding available for firms to establish advanced uctor Shorterm Longtam 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, ministriesof commerceor industry. Transportation Bioremediation Biofilms The intent of this new agency would be to Sanitation Bioremediation Sensors provide loans at favorablerates to small and medium businesses so they can hire the scientistsand buy the equipmentto perform other biotechnology relates to the industry the advancedresearch 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 remediationindustry is one other efforts to improve the business climate of the fastest growing industriesin 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 possibilityof firms the next five years is 25 percent (Chowdhury being able to importrequired technologyand 1992). Bioremediation is one of the tech- supplieswithout 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 componentthat aims The remediation industry is now concen- to improve on the ability of industry to inno- trated in industrialcountries. But it is certain vate. The setting up of an advancedresearch to spread to developingcountries, especially and development unit by a firm should ac- so since the technology is available, it is complishthis objective. relativelycheap, there are many opportunities The other PSD activity that is relevant to for local research to improve bioremediating the promotion of biotechnologyis its direct organisms and techniques, and there are fostering of private enterprise, including many situationswhere bioremediationcan be providing support for entrepreneurialefforts employedimmediately for great effect. (World Bank 1992). The Bank is involved in Another PSD-typeproject could be aimed many sectors where there are possibilitiesfor at establishingregional-based firms to raise adding value to raw material products via and maintainbroodstock and to enhanceseed biotechnology.This wouldincrease 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 logicaldependence on outsiders.Biotechnolo- industryrequires 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 initiativethat has environmentalimpli- hatcheries are in operation, each producing cations and that could employ marine and fewer than 40 million fry. Aquaculturists ExploringWorld Bank optlons 89

make up the difference by collectingfrom the potential to support projects that might seem wild, which is environmentally damaging. too risky for political or commercialreasons This situation is not unique; the major con- in developing countries. Capitalizingon the straint on mollusc aquaculture in Asia is the stabilitythat IFC provides in the development lack of seed (Lovatelli 1990). Making avail- of new commercialprojects, 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 developmentof marine ing broodstock would be environmentally biologicaljoint 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 competencein the field so nationalFinance Corporation(IFC), which is that it can properly assess the feasibility of the member of the World Bank group that biotechnologyprojects, thus minimizingthe supplies direct project financing for private risk of initial failures. investmentsin developingcountries, 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 appropriateassis- Developingthis 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 technologyis the most appropriatefor devel- could well become an element in projects to, oping countries because: entry into the field for example, increasethe resistanceof plants is easier and costs less than any of the other to diseases or pests, increase the ability of high technologies;many developingcountries crops to grow under arid conditions, design have an existingbase 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 biotechnologymay natural resources that can be exploited via be used in projects having health objectives biotechnologyfor sustainable,environmental- to, for example, to improve drugs or develop ly sound economicdevelopment; and certain vaccines;to expand industrialcapabilities by, problems facing populations in developing for example, utilizingagricultural wastes for countries related to disease, food supply, alcohol production; and so forth. Similarly, environmentaldegradation and energy supply projects aimed at assistingisland countries or may be amenable to technical solutions that countries with significant marine resources only biotechnologycan provide. In recogni- may include marine biotechnology compo- tion of the promises that biotechnologyholds, nents for objectivessuch as increasingyields scientistsand 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 respondedby sponsoringactivities point of the foregoing is that a Bank officer, designed to help enhance present capabilities when planning wide-rangingprojects on land in biotechnologyand 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 environmentsector, 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- sizeablebiotechnology components, and in its niques may have for reaching project objec- support of CGIAR that funds some aquacul- tives, somethingthat probably has not been ture and biotechnologyR&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 questionnaireswere 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 biotechnologyor 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 implementedby the section is devoted to one country. It begins Secretaria Ciencia e Tecnologia (SCT) by naming the ministry or other authority (Mr. Paulo Cesar GoncalvesEgler). 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. Departementode Biologia. changing. As a result, new departments are 3. Universidade Federal de Alagoas, being set up, old departmentsexpanded, and Macei6: marine biology. additional scientists are being hired to staff 4. Universidade Federal de Sergipe, them. Administrativechanges may have been Aracaju. instituted at local and national levels that a. Departementode 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 situationas it existed in late 1991. marine biology, marine pollution. Second, we were entirely dependent on 7. Instituto de Estudos do Mar Almirante informationsupplied by persons who head or Paulo Moreira, Arraial do Cabo: work at the institutionsthat are listed. Some macroalgae. of this informationmay represent wishes or 8. Universidadede 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 MARINEA90 TECHNOL OGY AND DEVELOPINGCOUNTRIES

9. Universidade Federal do Parana, Abeliuk) and the Comit6 Oceanografico Paranagud. Nacional(Mr. Hugo GorzigliaAntolini). a. Centro de Biologia Marina: marine In addition, Chile has a national biology, planktonstudies, bacteriobentos biotechnologyprogram 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 Universidadedo Rio Grande, Rio Grande. 1. Universidad Cat6lica de Valparaiso, a. Departemento de Biologia: Valparaiso. phytoplanktonstudies. 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. UniversidadeFederal 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. UniversidadeFederale do Santa Catarina, protein structure and enzymes. Floriandpolis. d. Escuela de Ingenierfa Bioqufmica: a. Departementode 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, antioxidantprotection extraction. of marine vertebrates, crustacean f. Laboratory at Vifia: macroalgae immunology, bacterial accumulationby tissue culture. mussels and oysters. 2. Universidad de Magallanes, Punta c. Departemento de Bioquimica: sea Arenas. anemonetoxins. a. Instituto de la Patagonia: d. Departemento de Zooligia: shrimp environmentalprotection. pathology. 3. Institutode Fomento Pesquero, Santiago (Dr. Patricio Bernal Ponce). Chile. a. Laboratoryat Putemdn(in cooperation with the UniversidadCat6lica de Chile): A. The authority having the main study and management of marine responsibilityfor marine biotechnology resources, includingmollusks and algae; or related areas is the Subsecretarfade induction o f mussel larvae Pesca (Mr. Andrds Couve) of the metamorphosis. Minesterio de Planificacion(Mr. Sergio b. Laboratoryat Coyhaique: aquaculture Molina). Other agencies having pertaining to salmonids. responsibilities in these fields are the 4. UniversidadCat6lica de Chile, Santiago: Corporacion de Fomento (Mr. Rend studies on macroalgae and pigment AppendixA 93

production. B. R&D Institutions. 5. Universidad Catolica de Chile, Sede Talcahuano, Talcahuano: macroalgae 1. Marine ScienceLaboratory, 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 responsibilityfor biotechnologyin 8. Lefersa Alimentos,Santiago: studieson 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. Responsibilityfor marineaffairs 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 DevelopmentAuthority Institute of Marine Sciedces and (Mr. Amitabh Kant), Ministry of Fisheries, Academy of Scientific Commerce, has responsibility for the Research and Technology, Ministry of developmentof 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. Divisionof 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 EnvironmentalAgriculture. bioremediation. 2. Goa University, Taleigo, Goa. Hong Kong. a. Departmentof Marine Biotechnology. b. Department of Marine Sciences (in A. Marine biotechnology appears to fall Bambolim): marine biology, marine under the purview of the Agricultureand microorganisms. Fisheries Department, Hong Kong 3. Central Marine Research Institute, Government. Cochin, Kerala. 4. Sri Paramakalyani College, Kallidaikurichi. 94 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

a. Post Graduate Department of B. R&D Institutions. Microbiology: screening of marine bacteria, algae and invertebrates for 1. Schoolof BiologicalSciences, 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 responsiblefor all science a. School of Marine Sciences: marine and technologyin 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. intertidalecology; ecologyof mangroves. a. Laboratoriode BiotecnologfaMarina: 9. Beshampur University, Beshampur, fish vaccines; detectionof Vibriospecies; Orissa. pollutioncontrol; fermentationby marine a. Department of Marine Sciences: bacteria. chemistry of the sea. 2. Centro de InvestigacionesBiol6gicas 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. Departementode Biotecnologfa,Instituto microbial enzymes;bioactive compounds de InvestigacionesBiomedicas - 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 responsiblefor all science halophilic microorganisms; andtechnologyistheFederalMinistryof bioremediation. Science and Technology (Dr. G. Ezekwe). Malaysia. B. R&D Institutions. A. Authorityto be determined. AppendixA 95

1. University of Port Harcourt, Port bioadhesion; biofouling; natural Harcourt. chemicalsfrom 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 Departmentof MarineBiology: 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 responsibilitiesfor marine biotechnology techniquesfor macroalgae. in China.TheAdministration for Science c. Invertebrate Laboratory: scallop and Technology(Mr. Song Chian) seems aquaculture; prawn aquaculture; to have major responsibilityfor scientific biofouling. research, while the Scientific and 3. Experimental Marine Biology TechnologicalSection (Dr. W.H. Yang) Laboratory, Academia Sinica, Qingdao, of the State Oceanic Administration is Shandong: research on transgenic fish; responsible for marine-relatedresearch. the constructionof a fish gene library; Further, the Department of Earth photosynthesisof 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 NationalAgriculture, Animal Husbandry shrimp and scallops; transgenicscallops; 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 Bioscienceand 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 preventionof fish diseases. biotechnologyremains 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 MARINEB0OTECHNOLOGYAND DEVELOING COUNTRIES

oysters, horseshoe crab, and others; pollution in ocean, macroalgac accumulationof heavy metals by fish. taxonomy. 8. ShanghaiFisheries University,Shanghai: 6. Universidad Nacional de Trujillo: fish studies on macroalgae. biology, invertebrateecology. 9. SouthChina Sea Instituteof 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 Investigacfonde Bioqufmicay Sinica: nitrogen fixation in cyanobacter. Nutricfon. 10. InstitutoTecnol6gico Pesquero: marine Peru. natural products, processing of marine living resources. A. The agency that is responsible for science and technology is the Consejo Philippines. Nacionalde Ciencia y Tecnologfa,while the Fondo de Reactivacfon del Sector A. The agency having responsibility over Pesquero (Ing. Luis Garayar Melendez) scienceand technologyis the Department of the Ministeriode 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. Institutodel 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. LearningResource Center: studieson 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 reproductionin 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 processingand 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. Fundamentalresearch in biotechnologyis research on seaweed species. the responsibility of the Life Sciences 6. AquacultureDepartment of the Southeast Division (Dr. Jung-Yaw Lin), National Asian Fisheries Development Center, Science Council, while the Iloilo: research on Gracilariabiology, 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: Councilof Agriculture. commercial production of carrageenan from the alga Eucheuma. B. R&D Institutions. 9. FMC Corporation: research on commerciallyvaluable macroalgae. 1. AcademiaSinica, 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 expressionof 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. Departmentof Aquaculture. seaweed stock improvement. b. Department of Microbiology. 4. Kaohsiung MedicalSchool. c. Departmentof BiologicalScience and a. Department of Microbiology: studies Technology. on gonyautoxins. d. Departmentof Fish Pathology. b. Department of Pharmacy: natural e. Departmentof Marine Biology. products from soft corals, monoclonal f. Department of Applied Chemistry. antibodiesfor gonyautoxins. g. Institute of Marine Sciences. 5. NationalChung 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. NationalDefense MedicalCollege. Biotechnology. a. Department of Microbiology and 1. EnvironmentalResearch Institute. Immunology: marine actinomycetes 2. Korean Ocean Research and exploitation. DevelopmentInstitute, Seoul. 7. National Pingtung Agriculture College: 98 MARINE BIOTECHNOLOGYAND DEVELOPINGCOUNTRIES

fish lymphocysticviral studies. 14. Tong Wu University. 8. National Sun Yat-sen University. a. Departmentof Microbiology: natural a. Departmentof Biology: luminescent products from marine bacteria. bacteria as biosensors. 15. Yang Ming Medical School. b. Department of Marine Resources: a. Departmentof Biochemistry: natural zoology of corals, natural products from products from marine bacteria. coral-associatedbacteria. 9. National Taiwan Normal University. Thailand. a. Departmentof Biology: fish hormone studies. A. Responsibilityfor marine biotechnology 10. NationalTaiwan Ocean University. appears to be split between two agencies. a. Departmentof Aquaculture: seaweed On the one hand, responsibility for the protoplasts, marine bacteria in nationalbiotechnology plan rests with the aquaculture, enzyme inhibitors from National Center for GeneticEngineering marine bacteria, transgenicprawn. and Biotechnology (Dr. Bhumiratana), b. Departmentof Food Science: marine Science and Technology Development toxins, natural products from bacteria. Agency. On the other, marine resources 11. NationalTaiwan University. fall under the authority of the a. Department of Agricultural Department of Fisheries (Dr. P. Chemistry: bioremediation,waste water Surasvadee),Ministry of Agricultureand 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 fermentationof 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 Edwardsiellaand Aeromonas,vaccine for substances; the study of heavy metal shrimp vibriosis, studieson adjuvantsfor resistancein microalgae. shrimp vaccine. 3. Department of Marine Science, f. Institute of Oceanography: seaweed Chulalongkorn University, Bangkok: biology, biology of soft corals, marine improvementof aquaculturepractices. thermotropic bacteria utilization. 4. Aquatic Resource Research Institute: 12. NationalTsing Hua University. managementof aquatic resourcesin fresh a. Department of Chemistry: natural and salt water, coastal zone resource products from seaweeds. management. b. Institute of Life Science: exotoxin 5. SrinakharinwirotUniversity, 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 Edwardsiellaand Aeromonas. Gracilaria. Appendix A 99

6. Brackishwater Fisheries Division, Uruguay. Kasetsart University, Bangkok: maricultureof black tiger shrimp. A. The ministry most concerned with 7. Faculty of Natural Resources, Prince of marinebiotechnology 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 Biologicaland Fishery Research Institute, Departmentof Fishery, Phuket: aquaculture and marine natural products development. Appendix B Special equipment requirements for advanced biotechnology

Equipmentrequirements for biotechnology Table B.1. Basicequipment for R&Dinclude optical microscopes, dissecting biotechnologyR&D scopes, centrifuges,incubators, sterilizers, tube and flask shakers, glass and plastic lio Apprvbatw price ware, chemicals,media, controlledlights, and growthchambers and greenhouses.The Computerfor sequence analysis S 11,000 more specialized marine biotechnology CO2 incubator 4,000 researchrequires aquaria(i.e., open ponds Digitizer 10,000 and closedtanks) togetherwith a seawater Electrophoresisseparation equipment 10,000 system for supporting it; and shops for Froezer(to -70° C) 6,000 mainWnng,rpairig and,as ned be, High-speedrefrigerated centrifuge 22,000 coaintaining,repairing and, as need be, Invertodphase microscope 10,000 constructingthis specialized equipment. Some Liquidair pumpwith a 180-liter research in marine biotechnologyrequires storagetank 18,000 sitingthe laboratoryclose to the ocean;other Reserch-gradedouble-beam work (e.g., naturalmarine products R&D) is Vpectrophotometer 28,000 not site dependent.All biotechnologyR&D Scanningdensitometer 18,000 institutionsshould provide strong support Scintillationcounter 30,000-35,000 facilities,such as libraries,electronic shops, Ultracentrifuge 50,000 technicalequipment service, information and computerservices. The material requirementsfor rDNA technologyare significant,as TablesB. 1 and B.2 demonstrate.Items listed in Table B. 1 Their reagent requirementsare also prodi- are thoserequired by a laboratorywishing to gious; for example,in the United Statesa take up plant biotechnologyresearch; the heavilyused automatedDNA sequencercan equipmentdemands for biomedicalresearch consumeabout $40,000 worth of expendable wouldbe greater.Those found in Table B.2 suppliesper year. are needed to perform exceedingly Monoclonalantibody work will require sophisticated research; in developing facilitiesfor culturinganimal cells, an animal countriessome items may be foundonly in a house, appropriateanimals, and the services nationalor regionalresearch institute. When of one or more animalhandlers. examiningthe two tables, keep in mind that The chemicaland specialsubstances needs equipmentupkeep requires a service unit of a biotechnology laboratory are mannedby skilled maintenanceand 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 equipmentwill soon break down United States, a researcher or technician and stay down. Further, each piece of requireschemicals and reagentswhose costs equipmentlisted in Table B.2 requires the range from $5,000 to $10,000 per year. servicesor a speciallytrained technician. Costsof theseexpendable 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 biotechnologyR&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 AutomatedDNA sequencer $100,000 storage. Flow cytometer 160,000 Equipment and supplies, while of lesser Oligonucleotidesynthesizer importance than highly trained scientific (manualmodel) 11,000 personnel, are essential ingredients to the Oigonucltotidesynthesizer carryingout 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 capabilitiesin biotechnology. Appendix C Definitions of marine biotechnology by scientists in industrial and developing countries

"Marine biotechnologyis the integration be called biotechnology,so one can applythis 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 Aurangabad431 004, India). industry, pharmaceutical industry, environmentalpollution and energy, medical "Marine biotechnology is a branch of diagnostics, fermentation industry, and marinescience 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). "Theterm biotechnologygenerally implies the applicationof technologyto organisms.In "Given the fact that biotechnologyis 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 biotechnologyis any aspect of biotechnology exploitationof 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 engineeringto marine sciences i.e. to utilize Center, Duke UniversityMarine Laboratory, the untapped gene pool in: North Carolina). 1. The transport of minerals (nutrient cycle) 2. Novel photosynthetic system (primary "Marine biotechnologycan 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) Universityof Science and Technology,Kochi 5. Marine pheromones, toxins, and 682022, India). pharmacologicalcompounds." (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 biologicalsystems, 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 biotechnologyfrom my understanding that: technologyto develop the methods for mass any marinebiological knowledge which could production and processing of marine be applied to increase yield or marine organismsfor a wide range of industrial and products is marine biotechnology. Marine commercial uses." (Dr. Saipin Chaiyanan, biotechnologyis 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 biologicalstructure and function on different level and artificially "I would say that marine biotechnologyis to control these variations by using the use of all the tools and knowledgein the engineering and technique, in order to life sciencesto produce a desired effect on or develop some new types of industry or new for mankind." (Dr. Robert S. Jones, biologicalproducts on a large scale, such as Director Marine ScienceInstitute, University genetic engineering,cell engineering,enzyme of Texas at Austin, Texas). engineering, microbial engineering, biochemicalengineering and the techniqueof "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 applicationson 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 definitionof marinebiotechnology is 843, Apartado Postal 2732, Ensenada, Baja the use of biotechnologyfor studies of marine California, Mexico). organismsor the use of marine organismsfor applications in the field of biotechnology." "Marine biotechnologyis the application (Dr. Bert Ely, Director Institute for of marine organisms including their systems Biological Research and Technology, or processes for the manufactureof 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 (MilagrosaR. Martinez, Associateprofessor biotechnologyas studies and developmentof and Director, Learning Resources Center, marine (aquatic)resources for human welfare University of Philippines at Los Banos, using the availablebiomolecular tools as well College, Laguna 3720, Philippines). as developingnewer and better research tools for application and improvement- "Marine biotechnology is the science enhancementof our understandingof marine dealing with the study of marine organisms (aquatic) life in general." (Dr. S.O. (preferentiallymicroorganisms 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 techniquesthat could be 104 WARINEBOTECHNOLOGYANDDEVELOPIN 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 marinebiological systemto etc) at a high quality and quantitylevel or to generate a desired product or products." degrade debris and undesirablesubstances in (Dr. Donald W. Renn, Senior Research by-products useful to mankind." (Dr. Fellow, FMC Corporation, Maine). Enrique C. Mateo, Fondode Reactivaciondel Sector Pesqueria, German Schereiber 198, "I shall define marinebiotechnology as the Francia 726 - Miraflores, Lima, Peru). commercial exploitation of living marine organisms or their components. The 'Marine biotechnologyas the application organisms will include microbes, and also of molecularbiological techniques/methods to plants as well as animals; the later will the production or modification of potential encompass the application of molecular commercialproducts. This might include the biology and cell culture techniques." (P.M. use of marine species for the application,or SatheeshSeshaiya, Post Graduate Lecturer in the use of molecular bio-techniques in the Microbiology, Post Graduate Department of marine environment." (Dr. DavidL. Nebert, Microbiology,Sri ParamakalyaniCollege, 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 applicationof the techniquesof biotechnologymanaged by the Institute is the modern molecularbiology to marinebiology. same used in other Latin America countries It covers the use of these techniquesto 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 commercialimportance. 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., technologywhere only DNA is manipulated." Massachusetts). (Dr. Patricio Bernal Ponce, Executive Director, Institutode FomentoPesquero, Jose Domingo Canas 2277, Casilla 1287, Santiago, Chile).

"Marinebiotechnology 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 DevelopmentBank AFDB AfricanDevelopment Bank AVHRR advancedvery high resolution radiometer BLI (BL2, BL3 or BL4) biosafetylevel 1 (2, 3 or 4) of the NIH guidelines C Celsius CGIAR ConsultativeGroup on IntemationalAgricultural Raerch COMAR Coastal and Marine Program CZCS coastal zone color scanner DNA deoxyribonucleicacid DLR Deutsche Forschungsfur Luft und Raumfahrt EBRD EuropeanBank for Reconstructionand Development E. coi Escherichia coU ECU Europeancurrency unit EIB EuropeanInvestment Bank ERS type of Europeansatellite EUREKA EuropeanResearch Coordinating Agency FAO U.N. Food and AgriculturalOrganization FDA U.S. Food and Drug Administration GABA gamma-aminobutyricacid GEF Global EnvironmentalFacility GESAMP Group of Experts on the ScientificAspects of Marin Pollution GIAM Global Impacts of AppliedMicrobiology GIFT genetic improvementof farmedtilapias GLOSS global sea-levelobserving system GMAG GeneticManipulation Advisory Group (United Xingdom) GOOS Global Ocean ObservingSystem HCMM heat capacity mappingmission hGH human growthhormone IARCs InternationalAgricultural ResearchCenters IBC InstitutionalBiosafety Committee ICES InternationalCouncil for the Explorationof tha Sea ICGEB InternationalCentre for GeneticEngineering and Biotechnology ICLARM InternationalCentre for Living Aquatic ResourcosManagement 1DB Inter-AmericanDevelopment Bank IFC InternationalFinance Corporation IGLOSS Integrated Global OceanSystem IHN infectioushematopoictic necrosis IMO InternationalMaritime Organization IOC IntergovernmentalOceanographic Commission JERS type of Japanesesatellite KFA type of Russian satellite LAL Limulus amebocytelysate LANDSAT Type of U.S. satellite MAB Man and the BiosphereProgramme MIRCEN MicrobialResource Center 106 MARINEBIOTECHNOLOGY AND DEVELOPINGCOUNTRIES

MSS multispectralscanner NCI U.S. National Cancer Institute NGO nongovernmentalorganization NIH U.S. National Institutes of Health NOAA U.S. National Oceanicand AtmosphericAdministration NRC U.S. National ResearchCouncil OECD Organizationfor EconomicCooperation and Developmnet OPEC Organizationof PetroleumExporting Countries PAN panchromatic PCR polymerasechain reaction PrepCom Preparatory Committee PSD private sector development RAC U.S. RecombinantDNA Advisory Committee rDNA recombinantDNA R&D research and development RFLP restriction fragmentlength polymorphism SAR syntheticaperture radar SLAR side-lookingairborne radar SPOT type of French satellite toH trout growth hormone TM thermal mapper U.N. or UN United Nations UNCED U.N. Conferenceon Environmentand Development UNDP U.N. DevelopmentProgramme UNEP U.N. EnvironmentProgramme UNESCO U.N. Educational,Scientific and CulturalOrganization UNIDO U.N. IndustrialDevelopment Organization U.S. or USA United States USDA U.S. Departmentof 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- combinationsto form peptides or proteins. isms, to make or modify products from raw Anabolism see metabolism. materialsfor agricultural,industrial or medical Anaerobic without oxygen. purposes. Antibody a specific protein moleculeproduced 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 substancethat when introducedinto 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 appropriatecircumstances 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 combiningnuclei and cytoplasmfrom 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 geneticallyidentical 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 experimentalor theoreticalwork 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 specificapplication. segmentof 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 ecologicalprocesses, Culture the growthof cells or microorganismsin which can be characterized at the genetic, a controlledartificial environment. taxon (for instance, familiesand species), and Dispersant a substance that reduces surface ecosystemlevels. tension of a floating pollutant, causing it to Bioremediation a technologythat uses biological sink. activity to treat contaminatedsoil or water in Database a collectionof data, defined for one or order to reduce or eliminate the more applications,which is physicallylocated contaminant(s). and maintainedwithin one or more electronic Biosafety in activities involving life forms or computers. their parts, the observanceof precautionsand Development the process of applying scientific preventive procedures that reduce the risk of and technicalknowledge to the practical adverse effects. 108 MARINEBIOTECHNOLOGYANDDEVELOPNG 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 concentrationsof slt. genetic information found in all living Hazard the likelihoodthat 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 ordinarycircumstane coded somewhere in an organism's com- of use. plement of DNA. Host a cell whose metabolismis used for growth Emulsant a surface-activesubstance that allows and reproductionof a virus, plasmid, or other a normallyimmiscible liquid (for exampleoil) form of foreign DNA. to disperse or become mixed into a second Host-vector system compatible host/vector liquid (for examplewater). combinationsthat may be used for the stable Enzyme a special protein produced by cells that introductionof foreign DNA into host cells. catalyze the chemical processesof 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- intestineand the intestinaltract of most other domaproduce large quantitiesa particulartype vertebratesas well. Somestrains are pathogen- of monoclonalantibodies. ic, causing urinary tract infections and Hydrocarbon one of a large and diverse group diarrhealdieas. Weakenedstrains are often of compounds,consisting of only carbon and used in laboratoryexperiments. hydrogen, constitutingpetroleum. Expression the translation of a gene's DNA Infection the invasionand sttling of a pathogen sequenceby RNA into protein. within a host. Fermentation the anaerobic bioprocessin which Intellectual property the area of law yeasts, bacteria or molds are used to convert encompassing patents, trademarks, trade a raw materialinto products such as alcohols, secrets, copyrights, and plant variety acids or choeses. protection. Filterfeeder an organismthat obtains its foodby 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 recoveringsuspendod organisms. an artificial environment, usually the Filterfeeders include baloen whales, corals, laboratory. musselsand sponges. In vivo literally winthe living'; pertainingto bio- Finfish true fish, as opposed to shellfish. logical processesor reactions taking place in Fraction a chemicalagent or compoundthat 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 physiologicalprocesses in a living organismin substances. which foodstuffare synthesizedinto complex Gene the fundamental unit of heredity. biochemicals (anabolism); complex Chemicallya gene consistsof ordered nucleo- biochemicals transformed into simple tides that code for a specificproduct or control chemicals (catabolism), and energy is made a specificfunction. 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 substancevital to the metabolismof 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 injectionof DNA into a cell living coll enabling it to produce more or or cell nucleus using a fine needle under a different chemicals. These techniquesinclude microscope. chemicalsynthesis of genes, the creationof re- Microorganism a microscopicliving entity that combinant DNA or recombinant RNA, cell can be a virus, bacterium, or fungus. Glssawyof 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. regulatoryaction. Decisionsusually 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, politicaland antigen. social factors. Nucleotide the fundamentalmolecule 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-sugargroup mentaryprotein. 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 crustaceanor mollusc. phosphate-sugargroup ribose. Synthesis the production of a compound by a Pathogen an organism that cauws diseas. living organism. Plankton microscopicorganisms inhabitingsea 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 (microscopicanimas). products or gnerate data. P,asmnid small, circular, self-replicatingforms of Toxicity the quality of being poisonous or the DNA often used in rDNA experiments as degree to which a substanceis 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. manipulationto 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 conversionof raw materialsinto bacteriophage(a virus that infects bacteria). products or components thereof through a Transfection the process in whicha bacteriumis series of manufacturingprocessos. modifiedin a way that allows the cell to take Real fime a characteristic of a system which up purified, intact viral or plasmid DNA. makesinformation available about a processso Transformation the introductionof new genetic quickly it allows the operator to act to change informationinto a cell using naked DNA (that the outcome of the process while it is still is, withoutusing a vector). under way. Triploid having three haploid sets of Recombinant DNA (rDNA) tho hybrid DNA chromosomesin each nucleus. resultingfrom the joining pieces of DNA from Vector a trsnsmissionagent, usually a plasmidor different sources. virus, used to introduce foreign DNA into a Risk the probabilityof injury, disease or death host cell. for persons or groups of persons undertaking Virs an infectiousagent, containingeither 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

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