Bioprospecting

Nicola´s Mateo, Werner Nader, and Giselle Tamayo Instituto Nacional de Biodiversidad (INBio)

I. Bioprospecting: A Tool for Survival and a Source tion created in 1989 to generate and disseminate of Inspiration and Innovation knowledge and promote sustainable uses of biodi- II. Economic Value and Beneﬁt Sharing versity. III. Concepts and Practices of Bioprospecting: The Case of Costa Rica

BIOPROSPECTING IS THE SYSTEMATIC SEARCH for GLOSSARY genes, natural compounds, designs, and whole organisms in wild life with a potential for product develop- bionics Science of systems, which function in the same ment by biological observation and biophysical, bio- way as or similar to living systems. chemical, and genetic methods, without disruption to biopiracy Illegal appropriation or exploitation of ge- nature. netic and biochemical resources. chemical prospecting Search for natural compounds in wild living plants, animals, and microorganisms with a potential for the development of chemical I. BIOPROSPECTING: A TOOL FOR products like pharmaceuticals, pesticides, cosmetics, SURVIVAL AND A SOURCE OF or food additives. combinatorial chemistry Laboratory methods to pro- INSPIRATION AND INNOVATION duce all possible combinations from various sets of chemical building blocks in a short period of time A. Traditional Bioprospecting and to generate molecular diversity for the screening In 1991 a 5300-year-old mummy was discovered in of new bioactive compounds. the Tyrolean Alps. The ‘‘Ice Man’’ and his effects were gene prospecting Search for genes in wild living plants, extremely well preserved and his clothing and equip- animals, and microorganisms for the breeding or ment were perfectly functional for the survival under genetic engineering of plants, animals, and microor- the harsh climatic conditions of the Alps. They con- ganisms in agriculture, fermentation, and cell culture sisted of plant and animal ﬁbers, oils, and waxes, put for agricultural and industrial production. together by a highly skilled and sophisticated Neolithic INBio The Instituto Nacional de Biodiversidad (Na- culture. The ‘‘Ice Man’’ suffered from an intestinal endo- tional Biodiversity Institute), a Costa Rican associa- parasite, the whipworm Trichuris trichiura, but he was

Encyclopedia of Biodiversity, Volume 1 Copyright  2001 by Academic Press. All rights of reproduction in any form reserved. 471 472 BIOPROSPECTING already equipped with the corresponding medicine, the In recent times, agronomists and professionals from fruiting body of Piptoporus betulinus, which contains related sciences have done considerable agricultural oils that are toxic to parasites and compounds that act prospecting looking for landraces having agronomic as strong laxatives that would cause expulsion of the advantages, particularly higher yields, pest and disease dead and dying worms and their eggs. resistance, and adaptation to specific microhabitats. The search for and utilization of biological resources Gene prospecting of the resources managed, nurtured, is as old as humankind and has been key to the survival, and selected over centuries by indigenous communities adaptation, and evolution of the human species. The has yielded considerable benefits to the world in terms perceptions and values of communities and individuals of improved crop varieties and hybrids. Traditional depending on and living in close proximity to wild plant breeding has slowed down in tropical countries and domesticated biodiversity are clearly different from due to lower research investment and unsuitable intel- those of the modern scientist engaged in bioprospect- lectual property protection, and this could diminish the ing. To many communities, biodiversity entails magic, opportunity of using the tools of the new biotechnol- religious, and ceremonial connotations in addition to ogies. the more mundane food, shelter, and medicine benefits Agricultural prospecting has resulted in the assem- derived from its domestication and utilization. bling of government, international, and private gene The forest has provided nourishment and shelter to collections, however, the viability of these resources, many cultures through a myriad of products and ser- mostly in tropical countries, has decreased over time. vices since early times. The Neotropical forests harbor Some estimates suggest that over 70% of all germplasm the wild ancestors of such major food crops as manioc, collected since 1940 is no longer viable or the associated cacao, and yams. Evidence of squash (Cucurbita knowledge has been lost. In situ conservation, therefore, moschata) farming in southwestern Ecuador over has become an attractive proposition to protect and 10,000 years ago suggests that agriculture in the Ameri- utilize the remaining genetic resources. cas may have begun in the rain forests (Piperno and Even plant breeding no longer depends only on genes Pearsall, 1998). Low population pressure and markets from wild types and ancestors of modern crops; new limited to exchange and barter enabled the original improved varieties can be artificially generated through indigenous groups to lead a low-impact lifestyle. Hunt- genetic engineering, for example, with designer genes ing and gathering, plus domestication of common that confer resistance to pests. Motifs in the amino acid beans, maize, cacao, chilies, squash, and other crops, sequences of the bactericidal and fungicidal magainin permitted not only the survival of autochthonous peptides from frog skin inspired genetic engineers to groups but also their social and cultural development, design resistance genes, which have scarcely anything and, quite significantly, with a minimal impact on the in common with their natural ancestors (Nader and natural environment. The arrival of Europeans in the Hill, 1999). Americas brought along agricultural practices that were to a great extent destructive to Neotropical ecosystems. These new farmers felt compelled to ‘‘conquer’’ nature B. Modern Bioprospecting and cleared increasingly large areas of the forest to plant Modern science and technology have provided substi- subsistence crops. Both systems have survived to the tutes for some biological products: nylon, polyacryl, present, with the latter practices and large-scale, chemi- and polyester instead of cotton and wool; aniline dyes cal-intensive agriculture being dominant because of in- instead of natural colorants; and organophosphates or creased population pressure and the need for higher carbamates instead of plant-derived natural pesticides agricultural productivity. like rotenone, nicotine, and ryanodine. Yet human inge- Costa Ricans, like other inhabitants of Middle nuity still depends in great part on inspirations from America, have used and continue to use medicinal wild life-forms to create these substitutes. In contrast plants (e.g., roots of Cephaelis ipecacuanha and Smilax to our forest-dwelling bioprospecting ancestors, we no spp.), fruits and nuts, palms, birds, animals, felines longer need to use the biological original to create a final (for hunting pleasure and to protect domestic animals), product; rather, through the study of these originals ornamental plants, feathers, and building materials we are able to discover and unveil the principle of a from the forest. The impacts of these practices, particu- biological function and then develop novel products on larly logging over the last 50 years, have been significant this basis. in altering the landscape and call into question the Millions of years of evolution have created a wealth tropical region’s capacity for sustainable development. of structures and mechanisms at the molecular, cellular, BIOPROSPECTING 473 and macrostructural level, all of which function eco- principle of flight was reduced to the vault of the wings. nomically and interact to perfection. Nature provides Vaulted wings derived from nature became the basic solutions to most of life’s technical problems. Natural structural feature of all airplanes (Hill, 1997). Given selection has imposed on living organisms the Min- the three inspirational resources from living nature— Max Principle: a minimum of material and energy ac- chemicals, genes, and designs—bioprospecting can be complishes a maximum of efficiency and stability. This differentiated into chemical, gene, and bionic prospect- makes biological prototypes particularly important for ing (Fig. 1). To discover new biological prototypes, our future given the world’s decreasing resources and bioprospectors can evaluate traditional uses of plants, increasing environmental problems (Hill, 1997). animals, and microorganisms, use random sampling, or, in the biorational approach, systematically analyze biological phenomena (Tamayo et al., 1997). Box 1 Biodiversity offers three fundamental sources of 1. Chemical Prospecting inspiration to the modern scientist: chemicals, The major defense mechanisms of plants against herbi- genes, and designs. Fields of applications include vores rely on chemicals: ‘‘The world is not colored green drug development, agrochemistry, and cosmetics to the herbivore’s eyes, but rather is painted morphine, (chemicals), development of recombinant phar- L-DOPA, calcium oxalate, cannabinol, caffeine, mustard maceutical proteins, enzymes, and agricultural oil, strychnine, rotenone, etc.’’ (Janzen, 1975). The co- biotechnology (genes), and architecture, mechan- evolution of herbivores and their feeding plants has ical engineering, and sensor technology (designs) created diversity on both the species and the molecular (Nader and Hill, 1999). levels (Ehrlich and Raven, 1964). Communication, competition, sexual attraction or rejection, and pollina- tion are also based to a great degree on chemistry and have contributed to the development of diversity. To An example of how nature inspires inventors is the help conserve this chemical wealth for future genera- development of the first glider airplane. By analyzing tions, Thomas Eisner developed the concept of chemical the flight of the stork (the biological prototype), the prospecting, a collaborative effort among conservation- principle of airlift was discovered and the functional ists, scientists, the pharmaceutical industry, and biodi-

FIGURE 1 The three principal sources of inspirations from biodiversity and their applications. Product examples are described in the text (with variations derived from Nader and Hill, 1999). 474 BIOPROSPECTING versity-rich countries to develop products from bio- from wild plants, animals, and microorganisms. The diversity and to generate income for its conservation antihypertensive proline derivatives enalapril (Vasotec) (Reid et al., 1993). and lisinopril (Zestril) were derived from a peptide in Although human ingenuity has created a huge vari- the venom of the fer-de-lance (Bothrops jararaca or B. ety of chemical compounds with exceptional pharma- athrox). Diclofenac (Voltaren) belongs, like aspirin, to ceutical activity without inspiration from nature—like the group of nonsteroidal anti-inﬂammatory drugs, the anxiolytic benzodiazepines valium and librium— which were derived from the lead structure salicin from natural compounds and their derivatives are still of willow bark (Salix spp.). The antineoplastic paclitaxel central importance in drug discovery. Table I lists the (Taxol) was discovered in the screening program of the 30 best-selling pharmaceuticals (drugs and biologics) National Cancer Institute from the bark of the Paciﬁc in 1997. Thirteen were either derived from or developed yew tree (Taxus brevifolia). The antiviral drug acyclovir as the result of leads generated by natural products (Zovirax) was developed using prior knowledge of cyto-

TABLE I Ranking of Key Ethical Pharmaceuticals (Drugs and Biologics) by Sales Value (U.S. $) in 1997 a

Rank Product Product category Company U.S. $ (millions)

1 Losec/Prilosec Proton pump inhibitor Astra/Astra Merck 3749 2 Zocor Hypolipidemic Merck & Co. 3575 3 Prozac Antidepressant Eli Lilly 2559 4 Vasotec ACE inhibitor Merck & Co. 2510 5 Zantac H2-antagonist Glaxo Wellcome 2254 6 Norvasc Calcium antagonist Pﬁzer 2217 7 Claritin Antihistamine Schering-Plough 1726 8 Augmentin Penicillin antibiotic SmithKline Beecham 1516 9 Zoloft Antidepressant Pﬁzer 1507 10 Seroxat/Paxil Antidepressant SmithKline Beecham 1474 11 Pravachol Hypolipidemic Bristol-Myers Squibb 1437 12 Ciprobay Quinolone antibiotic Bayer 1378 13 Premarin Hormonal product American Home Products 1328 14 Mevalotin Hypolipidemic Sankyo 1311 15 Biaxin/Klaricid Macrolide antibiotic Abbott 1300 16 Novolin Human insulin Novo Nordisk 1290 17 Pepcid H2-antagonist Merck & Co. 1180 18 Sandimmun/Neoral Immunosuppressant Novartis 1166 19 Epogen Antianemic Amgen 1161 20 Mevacor Hypolipidemic Merck & Co. 1100 21 Imigran Antimigraine Glaxo Wellcome 1085 22 Procrit/Eprex Antianemic Johnson & Johnson 1070 23 Neupogen Colony stimulating factor Amgen 1068 24 Adalat Calcium antagonist Bayer 1063 25 Rocephin Cephalosporin Roche 1062 26 Prepulsid Gastroprokinetic Johnson & Johnson 1050 27 Zestril ACE inhibitor Zeneca 1036 28 Voltaren NSAID Novartis 1019 29 Zovirax Antiviral Glaxo Wellcome 951 30 Taxol Anticancer Bristol-Myers Squibb 941

a Source: Wood Mackenzie’s PharmaQuant, Edinburgh, U.K., Jan. 1999. Products derived from natural compounds from wildlife animals, plants, and microorganisms are printed in bold, and products derived from natural compounds are in italic. BIOPROSPECTING 475 sine arabinoside, which was isolated from a Florida sponge (Alan Harvey, pers. comm.). Lovastatin (Meva- cor) and its synthetic analog simvastatin (Zocor) are derived from the fungus Aspergillus terrestris, and anti- hypercholesterolemic drugs like pravastatin (Pravachol or Mevalotin) are natural compounds from Penicillium citrinum. Ceftriaxone (Rocephin) is a semisynthetic third-generation cephalosporin antibiotic (Cephalospor- ium sp.) and cyclosporin (Sandimmun/Neoral) is an immunosupressant cyclic peptide from Tolypocladium inflatum. Co-amoxiclav (Augmentin) is a combination of the beta-lactamase inhibitor clavulanic acid from the bacterium Streptomyces lavuligerus and the semisynthetic antibiotic amoxicillin (Penicillium or Aspergillus spp.). Finally, clarythromycin (Biaxin/Klaricid) is a chemical derivative of the classic antibiotic erythromycin from the streptomycete Saccharopolyspora erythrea. Random screening for new drugs from natural compounds comprises the collection of materials from plants and animals and the isolation of bacteria FIGURE 2 (A) The antihypertensive drug enalapril (Merck & Co.). and fungi in large numbers. Extracts are prepared (B) The antihypertensive drug captopril (Bristol Myers Squibb). from these materials and then exposed to screening batteries with automated bioassay systems to discover novel biological activities. The bioactive principles zyme in the mammalian bloodstream, which converts are then purified and characterized by bioassay- the peptide angiotensin I to the hypertensive hormone guided fractionation. Since a large number of bio- angiotensin II. Analysis of the functional principle of active compounds will be already known and thus this snake peptide at the molecular level and the discov- lack novelty, these need to be identified as early as ery of the bioactive lead structure (a proline in combina- possible in the discovery process by dereplication. tion with the carboxyl group of a glutamic acid) finally Even if a compound is in the public domain, it led to the synthesis of small molecules like captopril might still be patentable in connection with a novel and enalapril (the technical realization of a biological application. Another drug discovery route is provided principle). Thanks to drugs like the ACE inhibitors and, by chemical screening, which fits the demand of especially, these deadly snakes from the Neotropics, modern high-throughput screening for large quantities human deaths from stroke and heart attack have de- of purified compounds. Without knowing their biolog- creased by over 50% during the last two decades. ical activity, extracts are first fractionated into pure Many of today’s pharmaceutical industry multina- compounds and these are then exposed to the bio- tional corporations became industrial giants by produc- assays. ing and selling products derived from ethnobotanical Among the most notable developments in modern research. Prior to Bayer’s aspirin, American and Eur- medicine were acetylcholinesterae (ACE)-inhibiting asian peoples treated fevers, inflammation, and pain drugs like enalapril (Fig. 2a), captopril (Fig. 2b), lisino- with salicin-containing plants like willows and poplars. pril, and perindopril. These compounds were derived Novartis’ diclofenac, number 28 in the list of top-selling biorationally from a peptide in the venom of a deadly pharmaceuticals of 1997 (see Table I), was derived from tropical snake (Wyvratt, 1988). The fer-de-lance the same salicin lead. Other examples are stabilizing (Bothrops athrox or B. jararaca) is a Neotropical pit muscle relaxants like tubocurarine, alcuronim, and viper that kills its prey by causing a drop in blood pancuronium, which are routinely used during anesthe- pressure (in the context of pharmaceutical develop- sia. They were derived from curare, an arrow poison ment, this represents the observation of a biological prepared from toxic plants by indigenous people in the prototype). Hypertension is a major health problem Amazon and Orinoco valleys of South America. The in most industrialized countries (the formation of an anticancer drugs etoposide and teniposide are deriva- analogy between a medical problem and a biological tives of podophyllotoxin, a glycoside isolated from the phenomenon). A peptide in the venom inhibits an en- roots of various species of the genus Podophyllum. These 476 BIOPROSPECTING plants possess a long history of medicinal use by early American and Asian cultures, including the treatment of skin cancers and warts. Cholesterol-lowering drugs like lovastatin, simvastatin, and pravastatin now have medical competition from a yeast extract from fer- mented Chinese red rice. Asians use these extracts traditionally as a food additive and the yeast produces lovastatin naturally. Secondary metabolites from plants have been used traditionally in agriculture as natural pesticides, but they were pushed out of the market to a great extent following the advent of DDT. Other natural pesticides are nicotine from tobacco leaves, rotenone from roots of the genus Derris, quassin from the wood of Quassia amara, ryanodine from leaves, stems, and roots of the tree Ryania speciosa, azadirachtins from the Asian neem tree, and pyrethrins from the perennial and Chrysanthe- mum cinerariifolium, originally from Dalmatia and Mon- tenegro (Wink, 1993). However, the toxicity of chemi- cally synthesized pesticides like organophosphates and chlorinated bicyclic compounds soon became evident and derivatives of natural insecticides, fungicides, and herbicides have gained increasing importance since then. Examples are the pyrethroids, the fungicides kresoxim-methyl (Fig. 3b) and azoxystrobin (Fig. 3c), and the herbicide phosphinothricin. Pyrethroids are derivatives of the pyrethrins, which were used traditionally as insecticides. Kresoxim-methyl and azoxystrobin are derived from strobilurin A (Fig. 3a), a natural fungi- FIGURE 3 (A) The natural fungicide strobilurin A from the mush- cide that was discovered via biorational research on the room Stobilurus tenacellus. (B) The strobilurin class fungicide kre- mushroom Stobilurus tenacellus. Random screening led soxim-methyl (BASF). (C) The strobilurin class fungicide azoxystrobin (Zeneca). to the discovery of phosphinothricin in the culture supernatants of a streptomycete (Crouse, 1998).

2. Gene Prospecting is more efficient and safer for therapeutic application. The selection of various phenotypes (and thus genes) Consequently other blood-sucking organisms like ticks, has been used traditionally for plant and animal breed- mosquitoes, and hookworms are currently under inves- ing, and modern gene technology offers many new op- tigation in attempts to discover novel anticlotting, anti- portunities. Numerous products derived from genes platelet, and vessel-constricting principles. from wild life are already on or are close to entering One example of the value of traditional know- the market (Tamayo et al., 1997). ledge is the development of the recombinant pharma- A powerful protein to treat acute heart attack was ceutical protein hirudin, a potent anticoagulant de- discovered biorationally in the saliva of the common rived from the saliva of leeches (Hirudo medicinalis). vampire bat, Desmodus rotundus. While feeding on the Leech therapy has been used in traditional medicine blood of their victims, these bats release analgesics and in Europe and Asia for centuries as the only efficient coagulation inhibitors with their saliva, including the treatment of thrombosis, thrombophlebitis, and hyper- Desmodus Plasminogen Activator (DPA), which dis- tension. solves thrombolytic blood clots and allows clot-free Traditionally, humans have used enzymes for the drinking. Recombinant human tissue Plasminogen Ac- production of cheese, softening of leather, and fermen- tivator (tPA) has been approved as a therapeutic agent tation of tea and tobacco. In modern technology, en- against heart attack in the United States and Europe, zymes have gained increasing importance in chemical although researchers at Schering AG found that DPA engineering, in the processing of animal and human BIOPROSPECTING 477 food and textiles, as a component of laundry detergents, and fungi, they rarely get infected by these pathogens. and for the diagnosis of diseases. Gene prospecting This common knowledge led to the discovery of antimi- searches for enzymes with new substrate specificity and crobial peptides, the magainins, which can be geneti- physicochemical characteristics, and gene technology cally expressed in transgenic plants and which confer allows their production in nearly unlimited amounts at resistance to fungi and bacteria. Random screening of low cost. Today random screening can be applied in actinomycetes from the soil of Cameroon led to the the discovery process at high efficiency rates and with- discovery of phosphinothricin acetylase, which confers out the need to isolate and cultivate microorganisms. resistance to the herbicide Basta in transgenic crops DNA is isolated directly from water, soil, dung, carcass, like soy bean, rape, and cotton. Resistance to insects or compost, then randomly cloned into expression li- can be achieved by the expression of insecticidal pro- braries; the host bacteria are subsequently screened for teins from soil bacteria, like delta-endotoxin from Bacil- enzyme activity in high-through-put screening robots lus thuringiensis and cholesterol oxidase from Strepto- on microtiter plates. From the hosts, the cloned DNA myces sp. (Tamayo et al., 1997). can be directly amplified and sequenced, and the genetic As a consequence of their absorptive mode of nutri- information used to construct highly productive host tion, fungi are able to exploit an almost infinite diversity strains for the bioreactors. of nutritional microniches. Recent studies suggest that Hydantoinases generate D-amino acids, high-value considerable fungal diversity exists even in what would fine chemicals for the production of half-synthetic anti- be considered a homogenes substrate. For example, biotics and pesticides. Particularly useful are thermosta- Bills and Polishook (1994a) adapted the particle filtra- ble hydantoinases, which were discovered by random tion technique to maximize the diversity of fungi iso- screening of cultivated thermophilic bacteria from hot lated from a substrate. Whereas traditional isolation springs in Yellowstone National Park in the United techniques select for fast-growing species that utilize States. Yellowstone was also the source of Taq polymer- primarily simple carbohydrates, the particle filtration ase from Thermus aquaticus, a crucial component of the technique favors isolation of those fungi actively grow- polymerase chain reaction (PCR). Its inventor, Kary B. ing in the substrate. In their study, fungi were isolated Mullis, plucked T. aquaticus from the American Type from 1-ml particle suspensions of tropical rain forest Culture Collection, where the microbiologist Thomas leaf litter. From each sample, 80–145 different fungal D. Brock from the University of Wisconsin had depos- species were isolated and the four replicates of the same ited it in the late 1960s. Recently some users of PCR substrate yielded between 300 and 400 different species have replaced Taq polymerase by Pfu polymerase, of fungi. In a second study (Bills and Polishook, 1994b), which was isolated from Pyrococcus furiosis (‘‘flaming fungi were isolated from only one vascular plant host, fireball’’) and works best at 100ЊC. Proteases and lipases Heliconia mariae J. D. Hooker. From 0.8-ml suspensions from psychrophilic bacteria, which decay whale cadav- of filtered particles of leaves from four plants, they ers in the deep sea, are of interest for producers of isolated about 50–100 fungal species per leaf sample. cold-wash laundry detergents (Madigan and Marrs, In both of these studies, approximately 40–60% of the 1997). fungi isolated were rare or unidentifiable. Clearly the Food for poultry and pigs is increasingly processed fungi are far more diverse than is currently recognized, with phytases from soil fungi like Aspergillus niger. even by mycologists. This enables these monogastric animals to assimilate Metabolic pathway engineering also offers new ag- the phosphorous from plants and diminishes the ricultural opportunities. A gene for a 12-carbon thioes- amount of phosphorous that enters the environment in terase that transforms the common rape plant into a their manure. In the United States alone, the widespread producer of high-laurate canola oil has been discovered application of phytase could prevent 82,000 tons of in the undomesticated California bay (Umbellularia phosphate from entering the environment each year californica). This oil is a perfect substitute for palm and contributing to the eutrophication of rivers and kernel and coconut oil, which are high-value commodi- lakes. ties from tropical countries. Already 70,000 acres of Genes from wild plants, vertebrates, insects, and soil transgenic rape are under cultivation in the United bacteria are used to create transgenic crops that carry States and Canada. resistance to pests and herbicides. They can be discov- Developments like Laurical (the trade name for this ered biorationally, by random screening, or on the basis high-laurate canola oil) displace the production of trop- of knowledge from traditional uses. ical commodities to the farm fields of developed coun- Although frogs live in ponds infested with bacteria tries, or even possibly to industrial bioreactors, posing 478 BIOPROSPECTING a threat to the economic survival of millions of farm major resource is still untapped: the insect world of the families in less developed countries. In Africa alone, tropics. Studies by Hill et al. (in press) in Costa Rica $10 billion in annual exports are vulnerable to industry- on butterflies and grasshoppers have revealed a diversity induced changes in raw material prices and require- of new coupling mechanisms and led to the definition ments. Most developments in plant biotechnology have of ‘‘tropical bionics’’ as a new branch in engineering. been achieved with crops mainly cultivated in industri- Insects became the most species-rich group of eukary- alized countries. This fact reduces the likelihood that otes by their capability to adapt to ecological niches. farmers in developing countries will benefit from the They are not only excellent chemists and genetic engi- new agricultural opportunities of gene technology (Ta- neers, but also masters in mechanical engineering, light- mayo et al., 1997). weight construction, and sensor technology. Figure 4 shows some of the morphological and physiological 3. Bionic Prospecting characteristics of a butterfly. Because of their diversity, Architects, designers, and engineers use prototypes insects are an enormous reservoir of potential techno- from nature for construction and technical solutions. logical developments. Josef Baxton, after seeing the radial ribbing in the vena- Until recently, the development of technical solu- tion of the Amazonian water lily (Victoria amazonica), tions in engineering on the basis of biological proto- designed on this basis the dome of the crystal palace types was mainly driven by biorational observations. in London. The inventor of Velcro fasteners was in- For example, sharks have a rough skin, and common spired by the stickiness of the seeds of a common farm sense suggests that rough surfaces should increase the weed, the burdock (Arctium spp.). The development of resistance in water and thus slow the fish down. How- the echo-sounder was inspired by the ultrasonic orien- ever, sharks swim at speeds of over 50 km/hr. This tation system of bats, and infrared sensors were based paradox led to the discovery that the structure of the on the design of the thermosensitive pit organ of the shark squamae impedes microturbulence, which com- rattlesnake. The latter developments are of particular monly emerges on smooth surfaces at high speed. The importance for military use and thus it is not surprising technical development of this principle led to the cre- that the concept of ‘‘bionics’’ was created in 1958 by ation of new surfaces for airplanes that significantly the Air Force major Jack E. Steele as the ‘‘science of reduce kerosene consumption. Another example of the systems’’ that function in the same way or similar to biorational approach comes from the lotus, considered living systems (Hill, 1997). to be the flower of purity and cleanliness in Asian cul- The discovery of new technical principles in living tures. Tiny wax papillae on its surface act as a self- systems is far from being completed, and at least one cleaning mechanism. This principle is currently being

FIGURE 4 The butterfly: a bionic system and technical prototype (with variations derived from Hill et al., in press). BIOPROSPECTING 479 used to develop self-cleaning surfaces for buildings In the highly competitive drug market, innovation and and cars. production speed are paramount, and consequently in A new tool for the discovery and development pro- the United States R&D costs increased within ten years cess in bionics was recently developed by Hill (1997). from U.S. $6 billion to $20.6 billion in 1998. The supply Structures and mechanisms discovered in nature are of screening batteries for the synthesis and extraction systematically listed in catalogs and organized after the of new compounds consumes 11% of these R&D bud- principal functions in engineering: connection, separa- gets and requires a workforce of 4400 professionals or tion, formation, carriage, support, transmission, storage 12.9% of all employees in R&D (Thayer, 1998). To and blocking of materials, energy, and information. compete with synthetic and combinatorial chemistry These catalogs of inspirations are available to the engi- efforts, bioprospecting needs to provide large numbers neer to screen for technical solutions, the blueprints of of natural compounds at high speed, for example, by which nature has already provided. chemical screening, or untap new and unusual resources. Global sales of U.S. $30 billion in 1995 makes agrochemistry the second largest biodiversity-related mar- II. ECONOMIC VALUE AND ket. Because of the toxicity and environmental problems BENEFIT SHARING associated with synthetic chemicals, pesticides derived from natural compounds currently dominate the mar- A. Biodiversity-Related Markets ket (see earlier discussion), however, development of resistance in pests continually requires new and innova- Markets related to products derived from biodiversity tive products and creates demand for bioprospecting. are significant, as shown in Table II. This table does Crop resistance to pests can be achieved by genetic not include bionic applications like sensor technology engineering and this might, at least partially, substitute and robotics. for some pesticide use in the future. The market for The estimated growth of the world drug market is the seeds of these genetically engineered varieties is about 6% per year, from 1997 sales of U.S. $295 billion estimated to grow to U.S. $50 billion in the year 2000. to $378 billion in the year 2001 (Thayer, 1998). To Currently most genetic transformation is done with become a billion-dollar product, a newly developed genes coding for insecticidal proteins from Bacillus thur- pharmaceutical has to be better than similar products ingiensis. However, it is only a matter of time before already on the market or reveal a new mode of action. crop resistance is overcome by pests, and therefore the discovery of new resistance genes from wild sources has become a central focus of bioprospecting efforts. TABLE II Other markets related to biodiversity are gaining Markets of Various Products and Services increasing importance, like those for enzymes (see ear- Market U.S. $ (billions) lier), phytopharmaceuticals, seeds, and natural cosmetics. The worldwide market for phytomedicines derived Drug market (1997) a 295 from traditional medicinal sources is estimated at U.S. Natural product-based drugs (1993) b 50 $12.4 billion, headed by products derived from ginseng, Phytomedicines (1993) c 12.4 ginkgo, garlic, horse chestnut, and echinacea (Gru¨ n- Pesticides d 30 wald, 1995). Nutraceuticals and functional foods offer Seeds (estimate for 2000) d 50 a health advantage in addition to nutrition. Examples Horticulture (U.K., 1991) b 1.6 are Chinese red rice and cholesterol-lowering oils, the Enzymes (1996) e 2.5 latter enriched in plant sterols and stanol, both by- Cosmetics (U.S.A., 1994) b 20 products of the paper pulping industry and corn wet- Natural cosmetics b 0.5 milling operations. With an estimated market of $7.9 Tourism (1995) b 3400 billion in 1997, these products are expected to compete Nature tourism (1988) b 12 effectively with cholesterol-lowering drugs like lovastatin. In general, the future market looks bright for a From Thayer (1998). products derived from nature, with an increasing de- b From ten Kate (1995). c From Gru¨ nwald (1995). mand for natural compounds and wild genes for use d From K. ten Kate, pers. comm. in genetic engineering. The same is true for bionics. e From Madigan and Marrs (1997). Recently, attempts have been made to put price tags 480 BIOPROSPECTING on natural resources, including biodiversity (Constanza ate economic growth at the community level and to et al., 1997). The value of sources of unique biological create incentives for conservation. materials and products was estimated as U.S. $79 per Economic benefits at the community level could in- hectare of forest per year. The value of yet undiscovered clude: growing, grading, and packing of medicinal pharmaceuticals in tropical forests was estimated at U.S. plants; cultivation of new ornamental plants; domesti- $3–4 billion for a private pharmaceutical company, and cation, cultivation, and primary production of biopes- as much as U.S. $147 billion dollars to global society ticides; cultivation and processing of aromatic teas; and as a whole. Although significant, these numbers do not exploration and evaluation of local landraces with resis- reflect the real value of inspiration that can be derived tant genes of interest. In all cases, a critical prerequisite from wild plants, animals, and microbes. Biodiversity for engaging in added-value biodiversity development provides the goods and services that are essential in at the community level is market research. Unless a supporting every type of human endeavor and thus clear market is identified, proper contracts are signed, enables societies to adapt to different needs and situa- and organization at the local level is achieved, there is tions. Biological prototypes may be crucial for the sur- no sense in proceeding with bioprospecting activities. vival of mankind in the future, and the ‘‘Great Hunger’’ in Ireland in the mid-1800s taught a bitter lesson. This potato late blight (Phytophthora infestans) caused the C. Biopiracy death of 1 million and emigration of another 1.5 mil- Our world would be a totally different place today if lion people. crops and domestic animals had not been moved from their centers of origins and domestication to other parts B. Bioprospecting and Conservation of the world. Coffee to the Americas, potatoes to Eu- rope, corn to Africa, and wheat to China are just a few The initial meaning of prospecting was the search examples of how germplasm has become the ‘‘patri- for precious metals and oil in the soil and subsoil. mony of humanity.’’ The ratification of the Convention Then Thomas Eisner, from Cornell University, coined on Biological Diversity (CBD) and the development the term ‘‘chemical prospecting,’’ and more recently and implementation of property rights regimes have bioprospecting has been used to indicate the search changed completely the prior existing scenarios on the for new sources of chemical compounds, genes, pro- spread and use of germ plasm. teins, microorganisms, and other products that have Today, in many cases genetic and biochemical re- economic potential and that can be found in the sources are jealously guarded and are the cause of con- world’s diversity of plants, animals, and microor- flict (when appropriate and legally sound negotiations ganisms. are absent) in many parts of the world. The term biopir- For bioprospecting to be an ally of conservation, it acy has been coined to reflect the illegal appropriation must be based on sound principles and criteria, and or exploitation of genetic and biochemical resources. must be linked to the concept of value. It was high- It must be added, however, that the lack of proper codes lighted earlier that the present and potential value of of conduct, lack of legislation, and the lack of national bioprospectible designs, genes, and chemicals is enor- capacity to handle biodiversity utilization issues in the mous and justifies the use of intelligent conservation majority of biodiversity-rich countries are factors en- strategies. To be effective as an element of conservation, couraging biopiracy. Even national researchers, when bioprospecting must contribute to the generation of dealing with poorly defined or cumbersome legal proce- wealth through R&D and must also alleviate poverty dures, tend to take shortcuts in their quest for useful in biodiversity-rich regions. In this context, biopros- genetic and biochemical resources. pecting programs at the national level can help to develop national capacity and provide economic returns to conservation projects if investments are made in D. Benefit Sharing Models and science, technology, and market research. However, this may not be sufficient to achieve sustainable devel- Bioprospecting Experiences opment: communities inside or bordering protected Perhaps the most controversial issue related to biopros- areas must participate in conservation and must benefit pecting in tropical developing countries involves benefit from bioprospecting activities in their own regions. The sharing. This is true for the case of the Instituto Nacio- utilization of biodiversity linked to added value at the nal de Biodiversidad (INBio) in Costa Rica, as well as local level appears to be a promising avenenue to gener- for other models and experiences developed elsewhere. BIOPROSPECTING 481

The central questions are: What is equitable? Who ben- assay-guided fractionation and only a handful of them efits and how? Who makes the ‘‘rules’’? will be pursued to complete structure elucidation. The issue of equitability is controversial and first Known natural products are quickly identified via estab- arose with the initial perception that millions of dollars lished dereplication. would flow back to the countries, organizations, and Benefit sharing agreements go back to the year 1986, individuals involved in bioprospecting. The experiences when the National Cancer Institute started its second of the last few years indicate that monetary benefits plant acquisition and screening program. Through con- (unless royalties would materialize) to host countries, tracts with the Missouri Botanical Garden, New York although significant, are limited in comparison to other Botanical Garden, and the University of Illinois at Chi- less tangible benefits such as technology transfer, in- cago, collections in over 25 countries were made under creased local scientific expertise, improvements in the Letter of Intent and later the Letter of Collection, legal frameworks, and enhanced negotiating capacities. in which the NCI unequivocally stated its intent to deal These less tangible benefits may be poorly understood with source countries in a fair and equitable manner. or underappreciated by some segments of society, who By inviting source country scientists to the NCI labora- quite rightly are interested in achieving a direct flow tories, these agreements include a technology transfer of resources and economic benefits to the local commu- component as well as a commitment of the Institute to nities living near conservation areas. require successful licensees of their anticancer drugs to Conditions are different in every country and there- enter into further benefit sharing agreements with fore local solutions should be tailored to local circum- source country government agencies or organizations. stances. Inside Costa Rica’s governmement-protected Furthermore, license applicants are required to seek as wildlands, for example, there are no human settle- the first source of supply the natural products from ments, in contrast to the case in many other biodiver- the source country. The letter also acknowledges the sity-rich countries, and therefore straightforward achievements and intellectual property rights of local agreements developed by INBio with the government, collectors, taxonomists, and healers. In a further devel- the academic sector, and private companies do not need opment of its benefit sharing policy, for several years to address the more sensitive issues of indigenous the NCI has negotiated Memoranda of Understanding knowledge and direct monetary compensation to com- (MOU) with 13 source countries, including Costa Rica. munities. Current bioprospecting experience, such as These MOUs are cooperative research agreements and in Costa Rica, indicates that benefits to society at large recognize that source country scientists and organiza- include an eanced knowledge of the country’s biodiver- tions are committed to performing an increasing share sity and improved information dissemination to various of the operations in-country, as opposed to the export sectors of society through guidelines and other publica- of raw materials. tions, information available on the Internet, national A successful example of this benefit sharing policy and international workshops, and education programs. is the company Sarawak Medichem Pharmaceuticals, This scenario is a significant improvement to the infor- Inc., which was founded in 1996 as a joint venture mal arrangements that existed prior to the CBD, by between Medichem Research, Inc., a small pharmaceu- means of which individuals would supply germplasm tical company based near Chicago, and the Sarawak samples to companies without any responsibility or state government. Sarawak Medichem is developing (ϩ) compensation to society. calanolide A as an anti-HIV drug. Calanolides are potent Perhaps one of the more comprehensive and system- inhibitors of retroviral reverse transcriptase and were atic bioprospecting programs has been the one carried originally found in leaves and twigs of the tree Calophyl- out by the National Cancer Institute (NCI) of the United lum lanigerum, collected in Sarawak, Malaysia, in 1987, States. As of 1959 the NCI had major contract-sup- through the NCI contract with the University of Illinois ported programs to collect, identify, and screen plant at Chicago. products and fermentation broth. A modest marine ac- Australia is actively involved in several bioprospect- quisition program was established in 1972. Central ob- ing initiatives and has made significant attempts to add jectives of the NCI have been cancer and more recently value and advance product development in the country. HIV research. Natural product extracts and other sam- It has also developed strategic partnerships with South- ples are screened in vitro against panels of human tumor east Asian and South Asian countries to increase the cell lines and against HIV-infected cells. Acquisition of availability of germplasm resources. Selected initiatives samples concentrates on marine and unusual microor- include those of AMRAD to do bioprospecting on the ganisms. Active natural components are isolated by bio- flora of the Northern Territory and the Australian 482 BIOPROSPECTING

Antartic Territory, AIMS focusing on marine samples of the shaman-identified plants were bioactive. This around the coast of Oz in partnership with several orga- small difference is to be expected, given that the assays nizations, research on the flora of Queensland by local are not specific to how the shamans use the plants. The laboratories linked to ASTRA, and a new laboratory ICBG Suriname project has also been able to describe based in Lismore, New South Wales, that will address, new plant species. This is relevant given that tropical among other areas, research and development on me- forests have been the source of 60% of the anticancer dicinal plants. drugs discovered in the last 10 years and offers a poten- The International Cooperative Biodiversity Group tially powerful economic reason for preserving the for- (ICBG) Program is an experimental program that ad- est, based on the $200 billion market for plant-de- dresses the interdependent issues of drug discovery, rived drugs. biodiversity conservation, and sustainable economic The experience of INBio as a participant in one of growth. Funding from the Institutes of the National the initial ICBG projects has been positive in terms of Institutes of Health (NIH), including the National Can- knowledge generation and capacity building, however, cer Institute, supports awards of approximately it also indicates how the role of the industrial partner $450,000 per year to the five initial initiatives. As of was not clearly defined nor negotiated at the outset. 1997, nearly 4000 species of plants and animals had This issue has been explicitly raised by Chapela (1996), been examined for biological activity in 13 different and in his opinion the five ICBG Consortia reflect the therapeutic areas. ICBG-supported projects include ac- specific interests of the academic partners with little quisition and analysis of natural products derived from consideration of the objectives of the industrial part- biological diversity as potential therapeutic agents for ners. The interest of industry in the ICBG may be more diseases of concern, to both developed and developing closely related to discovering microorganisms (which countries, like AIDS, malaria, tuberculosis and other were explicitly excluded) present in some of the sam- infectious diseases, cancers, central nervous system dis- ples to be collected (Chapela, 1996). orders, and heart disease. The isolation of active compounds from tropical The active ICGB projects have made variable prog- plants with a history of medicinal use was the main ress in knowledge generation, local capacity building, strategy of Shaman Pharmaceuticals in their efforts to training, and the identification of promising lead com- discover and develop novel products for human dis- pounds. The one in Suriname, briefly reported here, eases. The company founded the Healing Forest Con- involves the participation of the Virginia Technical In- servancy, a nongovernmental organization, explicitly stitute, the Missouri Botanical Gardens, Bristol Myers to develop and implement a process by which to return Squibb Pharmaceutical Research Institute, and the Na- benefits to countries and cultures that chose to contrib- tional Herbarium of Suriname. After conducting some ute to its drug discovery program. Shaman contributed 14,000 assays from 1993 to 1998 of more than 3300 up to 15% of its drug discovery costs to fund projects extracts, Virginia Tech has identified 30 unique extracts or programs that were based on the expressed needs of that have activity and has isolated 20 chemical com- the local communities. Shaman also provided medium- pounds that have bioactivity. The most interesting com- term reciprocity programs that included training for pounds discovered are a group of alkaloids from Eclipta national scientists at Shaman’s California laboratories, alba that have good antifungal activity, better in some equipment and resources to conduct research in host cases than the clinically used drug amphotericin B. countries, and the funding of studies to evaluate sus- However, these alkaloids also have weak cytotoxicity, tainable harvesting of indigenous medicinal plants. Ac- and the decision was thus made not to develop them cording to Shaman staff, the practical dilemmas of dis- as antifungal agents. tributing fair compensation based on intellectual Meanwhile, Bristol Myers Squibb has put more that property rights are numerous, and therefore Shaman’s 3000 extracts through 32 screens each in six therapeutic approach involved an upfront return and at each further areas, with the result that one promising compound stage of the product development process. The recent (for anticancer activity) is still being tested. While look- and significant reorganization of Shaman in early 1999 ing for potential pharmaceutical products, project sci- will likely affect the ongoing benefit sharing experiences entists conduct ethnobotanical and random botanical with the communities. collections using yeast-based assays. The Virginia Tech Leading botanical gardens have developed benefit researchers found there was a slight benefit to the ethno- sharing strategies to compensate the countries where botanical approaches: 2.8% of the plants collected at botanical specimens are initially collected. Through random were active in the yeast assays, whereas 3.8% their collaboration with the National Cancer Institute, BIOPROSPECTING 483 the New York Botanical Garden and the Missouri Botan- Other relevant initiatives to value biodiversity in- ical Garden voluntarily adopted policies on the research clude payment for environmental services such as CO2 of natural products in the mid-1980s. Most notably the fixation. A 5% fuel tax yields approximately $40 million Missouri Botanical Garden committed itself to enter yearly, which is invested in carbon sequestration pro- into commercial research agreements only with the programs to support the conservation of forests and refores- vision that royalties will be paid to the source country tation. Serious consideration is also being given to and that the Garden itself will not receive any percent- charge users for the actual costs of water and electricity, age of such royalties. In 1981 the New York Botanical and initial experiments are under way. Currently, water Garden established the Institute of Economic Botany bills reflect essentially distribution costs and not the to focus a portion of the Garden’s research enterprise value of this essential commodity to society. on applied botanical questions of great human concern. Costa Rica is considered to be a megadiversity coun- One of its forest projects is the Belize ethnobotany try. Most of its territory has a diversity density of over project, begun in 1988, to conduct an inventory of 5000 species of vascular plants per 10,000 km2. Only the diversity of medicinal plants and to preserve this 10 other countries on earth (Nicaragua, Panama, Co- richness. In Europe, the Royal Botanical Gardens at lumbia, Ecuador, Peru, Bolivia, Brazil, China, Malaysia, Kew took the lead and its benefit sharing policy also and Papua New Guinea) have areas with similar diver- includes those materials that had been collected prior sity (Barthlott et al., 1996). However, of the total num- to the ratification of the CBD. ber of species estimated for the country, only a minor A very innovative and comprehensive strategy is portion has been described. Vertebrates and vascular that of South Africa, where traditional healers, the plants have been well studied, with over 90% of the government, and private and academic sectors are species documented and characterized. For all other joining efforts to undertake bioprospecting. The initial groups of organisms, including insects, algae, fungi, investment (40% government and 60% private sector) and bacteria, there are enormous gaps of knowledge. of U.S. $10 million will fund the investigation of The management and development of biological re- 23,000 plant species plus marine organisms and micro- sources fall under the responsibility of the Ministry organisms. The plan is to develop international part- of Environment and Energy (MINAE), specifically the nerships, however, most of the R&D will be done National System of Conservation Areas (SINAC), which in-country. is the institution in charge of conserving and promoting the sustainable use of the country’s natural resources. The eleven Conservation Areas in which the country III. CONCEPTS AND PRACTICES OF has been divided constitute the various regions estab- BIOPROSPECTING: THE CASE OF lished by MINAE to carry out a decentralized manage- COSTA RICA ment of biodiversity and natural resources, with the participation of the communities surrounding the pro- A. Nature-Based Initiatives and the Role tected areas. Executive Presidential Decree No. 19153 of June 5, of the National Biodiversity Institute 1989, established a Biodiversity Planning Commission, Costa Rica has developed a system of conservation with representatives from various governmental organi- areas in which over 25% of the national territory is zations, higher education institutions, and nongovern- allocated to nature reserves and national parks. Both mental conservationist organizations. This Commission the government and civil society have a role in recommended the creation of the National Biodiversity conservation and have contributed to creating aware- Institute (INBio) as a nonprofit, nongovernmental orga- ness of the importance of biodiversity at the national nization to serve the public good. INBio was legally and international levels. In terms of economic benefits, registered on October 26, 1989, as a civil association Costa Rica derives its main source of income from governed by an Assembly of Founders and a Board nature-based tourism; this sector yielded over U.S. of Directors. $850 million in 1998 and appears to be growing INBio works under the premise that a tropical coun- steadily. Surveys indicated that 70–75% of all tourists try will be able to conserve a major portion of its wild come to experience nature. This is an impressive biodiversity if this biodiversity generates enough intel- development when compared to the traditional eco- lectual and economic benefits to make up for its mainte- nomic sectors of banana and coffee production $500 nance. INBio and the Ministry of Environment and million and $350 million, respectively). Energy established a collaborative agreement that 484 BIOPROSPECTING allows INBio, within the existing legal framework, to high school students, and information for visitors. This conduct biological inventories, biodiversity prospect- program also cooperates with various national commis- ing, and management and distribution of Costa Rican sions of private and public organizations. biodiversity information. One of INBio’s goals is to generate greater awareness of the value of biodiversity and, thereby, promote its B. Partnerships and Management Issues conservation and sustainable utilization based on the demands of national and international users. This goal Current bioprospecting approaches require a significant is fulfilled through the integration of the following pro- degree of partnership at various levels. In Costa Rica, cesses: INBio has developed a close alliance with the government (Ministry of Natural Resources and Energy) that • biodiversity inventory focused particularly on the makes bioprospecting development possible in the na- country’s protected wildlands; tion’s Conservation Areas. This first level of partner- • conservation for development; ship is essential and is based on trust, understanding, • identification and promotion of sustainable uses of and clear-cut responsibilities and benefits (see Section biological resources; III,D). • organization and management of information; and Another essential level of partnership involves the • generation and transfer of knowledge. academic sector. Universities, both local and international, complement the required expertise in biology, The Biodiversity Inventory Program is carried out chemistry, microbiology, and biotechnology that is cen- in Conservation Areas by ‘‘parataxonomists.’’ They are tral to the concept and need of adding value at the in charge of collecting, preparing, and mounting the national level. Last but not least, a strong alliance with biological material with pertinent field information, and the private sector (conservation, pharmaceutical, ag- local curators are responsible for the taxonomic classi- ricultural, biotechnology, and fragrance companies) fication of such material, with support from national allows market forces and the needs of society to deter- and international experts. mine the products and services to be pursued through The Conservation for Development Program helps bioprospecting. The country has a limited capacity to strengthen the capacity of INBio and assists in the tasks invest in science and technology, and therefore partner- assigned by the National Conservation Strategy, which ships with the academic and private sectors are the only is based on protecting, understanding, and using bio- way to accomplish many of the stated goals. diversity in a sustainable manner. The Biodiversity Information Management Program processes and organizes the information generated by C. Mechanisms and the Legal Framework the inventory and other national collections under vari- Managing the complexity of the institutional interac- ous formats, according to the needs of users. Thus, it tions and addressing the needs and expectations of the seeks to process and integrate texts, maps, photographs, various partners require careful planning and organiza- and quantitative information about Costa Rican biodi- tion. Costa Rica and INBio cannot provide a prescrip- versity. tion for bioprospecting success, but do offer 10 years Natural resources constitute an essential source for of experience full of drawbacks, challenges, and new medicines, agricultural products, coloring com- achievements. pounds, resins, and other uses. Through the Biopros- The requirements of a bioprospecting program are pecting Program and in partnership with national and relatively simple to state: vision, leadership, negotia- international organizations, INBio carries out a system- tion, planning, understanding partners’ strengths and atic search of chemical compounds, genes, and micro- expectations, teamwork, realistic research plans, and and macroorganisms that may help find new products sufficient local expertise in science and informatics. But for the benefit of humanity. these elements are not necessarily simple to operation- The need to disseminate biodiversity information alize. It is widely perceived that being a small country motivated the creation of the Social Outreach Program and having a well-educated workforce and the right as a means to distribute information to different sectors incentives make the process easier in a country like of society. It includes production and publication of Costa Rica. All this is probably true, however, there are printed and audiovisual material on biodiversity issues few comparisons available to ascertain the validity of for various users, special programs for elementary and this hypothesis. BIOPROSPECTING 485

TABLE III 2. Compensation The Legal Framework for Biodiversity In the INBio/SINAC strategy, compensation is negoti- Conservation and Bioprospecting in Costa Rica ated at two different levels: • Forestry Law, 1995 • National Parks Law, 1977 (a) All research costs related to a bioprospecting • Wildlife Conservation Law, 1992 agreement (e.g., collecting, GIS, natural history • Ministry of Environment Law, 1990 extraction, determination of activity, etc.) are • Convention on Biological Diversity (CDB), 1993 paid by the interested company. A contribution • General Environmental Law, 1995 equaling 10% of all operational costs is trans- • Biodiversity Law, 1998 ferred to SINAC for conservation purposes. (b) In case a product based on a sample provided by Costa Rica reaches the market, royalties are pay- A country’s legal framework, based on both theory able in lieu of patent rights. Current patent laws and experience, sets the fundamental organization in Costa Rica offer limited protection and are and incentives not only for conservation but also for therefore unsuitable for products destined for the bioprospecting. Table III highlights significant pieces world market. Royalties vary significantly, de- of legislation that have made conservation and utiliza- pending mostly on added value generated within tion of biodiversity possible in Costa Rica. The 1998 the country. Biodiversity Law, a law of consensus among the various stakeholders, reflects in a significant way Literature surveys indicate that 1–15% royalty ranges the country’s prior experiences in conservation and are the norm; INBio’s experience coincides in most bioprospecting. cases with these figures. The transfer of resources from bioprospecting agreements to research and conservation programs is shown in Table IV, and the list of D. Criteria active bioprospecting agreements (both academic and Out of the 45 articles contained in the Convention with industry) is given in Table V. on Biological Diversity (CBD), INBio and the National System of Conservation Area (SINAC) of the Ministry 3. Transfer of Technology of Environment and Energy have emphasized the fol- Access to partner companies’ research processes and lowing ones in the design, approval, and implementa- bioassays of interest to INBio and its local partners is tion of bioprospecting agreements. negotiated as part of the transfer of technology. INBio’s current research capacity in bioprospecting is largely 1. Access based on technology acquired through such agreements Signatory countries of the CBD have agreed to facilitate with the private and academic sectors. access to other members. INBio and SINAC comply with this principle provided that other agreed criteria 4. Training (presented in the following sections) are an integral Contributions that a host country can make in biopros- part of the access agreement. pecting research and product development are essential

TABLE IV Contributions and Payments from Bioprospecting Agreements (U.S. $)

1993a 1994 1995 1996 1997 1998 (Jan–Feb) Total

Ministry of the Environment and Energy (MINAE) 110,040 43,400 66,670 51,092 95,196 24,160 390,558 Conservation Areas 86,102 203,135 153,555 192,035 126,243 29,579 790,649 Public Universities 460,409 126,006 46,962 31,265 34,694 326,486 1,025,822 Other groups at INBio 228,161 92,830 118,292 172,591 129,008 0 740,882 Total 884,712 465,371 385,479 446,983 385,141 380,225 2,947,911

a Estimated amounts since 1991. Source: INBio, 1998. 486 BIOPROSPECTING

TABLE V INBio Bioprospecting Agreements

Partner Year started Application

University of Costa Rica 1991 General agreement 1998 Operation of an NMR unit University of Strathclyde (U.K.) 1991 New pharmaceutical products from plant sources Merck & Co. (New Jersey, USA) 1991/1994/1996/1998 Pharmaceutical and veterinary products from plants and microbial sources BTG/ECOS-Costa Rica 1992 Development of a bionematicide from Lonchocarpus sp. Cornell University 1993 Drug discovery Universidad Nacional, Costa Rica 1993 General agreement Bristol Myers Squibb/Cornell University 1994 Insects as source of new compounds for pharmaceutical industry Instituto Tecnolo´gico de Costa Rica 1994 General agreement Givaudan Roure (New Jersey, USA) 1995 Fragrances and aromas from Costa Rican biodiversity University of Massachussets 1995 New insecticides Diversa (San Diego, USA) 1995/1998 New enzymes from extremophilic and other microorganisms Government of Canada 1996 Debt for nature swap INDENA (Italy) 1996 New antimicrobials for dermatological use from plant sources EARTH/NASA/Other Latin American institutions 1997 Chaga Space Project : compounds against regulatory enzymes University of Strathclyde (U.K.) 1997 Pharmaceutical products Office for Scientific, Technical and Cultural Affairs 1998 MOSAICC Project (Belgium) Phytera (Massachusetts, USA) 1998 Pharmaceutical products from plant sources elements of any added-value strategy. In every the national biodiversity, transfer of technology train- agreement, one or two training sessions for Costa Rican ing, and development of negotiating capabilities. How- scientists and technicians per year (normally of short ever, we have also learned that bioprospecting is not duration) are negotiated at the company’s expense. the gold mine that was initially envisioned. There are no guarantees that royalties will materialize in the fu- 5. Sustainable Uses ture and therefore they should not be included in deci- INBio and its partners will not engage in extractive sion-making and budgetary processing. Obviously, if strategies should a forest resource become of interest they become a reality they will be a welcome incentive to the market. Instead, they will pursue chemical and support for conservation and research. synthesis approaches if possible, a domestication We have learned that partnerships and alliances are strategy if feasible and desirable. In the latter case, indispensable, that innovation must be at the forefront a species may be successfully domesticated and be- of our activities, and that controversy and misunder- come a new agricultural crop or a new productive standings are part of the daily routine. We have also enterprise with the potential to generate economic de- learned, in order for history not to repeat itself, that velopment. Costa Rica must develop internal R&D capacity leading to advanced product development processes and in cer- tain cases to finished products. Private companies have E. Lessons Learned, Challenges, shown a cyclical interest in natural products, and therefore bioprospecting has had and will continue to experi- and Opportunities ence peaks and valleys of demand. Costa Rica and INBio have gained considerably from At INBio, bioprospecting is only one component bioprospecting agreements, through knowledge about of a much wider national biodiversity strategy. The BIOPROSPECTING 487 major factors responsible for INBio’s initial accom- should be noted that many highly successful drugs, plishments have been solid political support, leader- which have been introduced to the market only re- ship and vision, a commitment to innovation, a diversi- cently, are derivatives of natural leads that were discov- fied portfolio of goals, and strategic alliances (Mateo, ered many decades ago. Examples are diclofenac (from 1996). Early bioprospecting efforts emphasized de- salicin), clarithromycin (from erythromycin), and cef- mand-driven agreements with pharmaceutical, bio- triaxon (from cephalosporin). technology, agricultural, and fragrance companies. In conclusion, the hunters, gatherers, and agricul- This line of activity will certainly be maintained and turalists of the Old and New Worlds are, in some enhanced, and it should evolve toward higher levels ways, the equivalent of the modern biochemical and of research and services that are performed within gene bioprospectors. Perhaps the methods and strate- Costa Rica (the added-value contribution). In parallel, gies differ, yet the goals are essentially the same: and given the particular conditions in Costa Rica, survival, well-being, and spiritual and material ad- INBio will also be involved in the development of new vancement. biodiversity products (nutraceuticals, phytomedicines, ornamentals, etc.) with national industries and local communities. This strategy will involve a pilot product See Also the Following Articles development phase followed by marketing of biodiver- AGRICULTURE, TRADITIONAL • BREEDING OF PLANTS • sity-derived products. Cost recovery mechanisms will ECOLOGICAL GENETICS • ECONOMIC VALUE OF be designed to support new product development ini- BIODIVERSITY • GENE BANKS • PLANT SOURCES OF DRUGS tiatives. AND CHEMICALS The recently born concept of modern bioprospecting Bibliography is already being challenged by new developments in science and technology. In an endeavor to reduce risks Barthlott, W., Lauer, W., and Placke, A. (1996). Global distribution and to increase the probability of success, companies of species diversity in vascular plants: Towards a world map of phytodiversity. Erdkunde 50, 317–328. are trying to substitute natural (incalculable) products Bills, G. F., and Polishook, J. D. (1994a). Abundance and diversity with artificial (calculable) products wherever possible. of microfungi in leaf litter of a lowland rain forest in Costa Rica. The success rate in drug discovery is estimated to be Mycologia 86, 187–198. about one product out of 10,000 screened chemicals Bills, G. F., and Polishook, J. D. (1994b). Microfungi from decaying or plant species for one field of diseases. To maintain leaves of Heliconia mariae. Brenesia 41–42, 27–43. Chapela, I. H. (1996). La bioprospeccioń en la era de la informacioń: 10% annual growth, or roughly $1 billion in new reve- Un ana´lisis crı´tico de las iniciativas de conservacioń asociadas nues, drug companies must bring two or three new con el descubrimiento de nuevos fa´rmacos. In Biodiversidad, Bio- products to the market each year, which is four to six tecnologıá y Desarrollo Sostenible en Salud y Agricultura : Conexi- times the current average achieved by the top produc- ones Emergentes, pp. 29–49. Organizacioń Panamericana de la ers. To speed up the discovery process, companies are Salud, Washington, D.C. Constanza, R., d’Arge, R., de Groot, R., Farber, S., Grasso, M., Hannon, investing in Ultrahigh Throughput Screening (UHTS) B., Limburg, K., Naeem, S., O’Neill, R. 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