From Forest to Fumaroles: a New Era in Bioprospecting ______

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FROM FOREST TO FUMAROLES: A NEW ERA IN BIOPROSPECTING ______

A Thesis

Presented to the

Faculty of

California State University, Fullerton ______

In Partial Fulfillment

of the Requirements for the Degree

Master of Arts

Geography ______

Curtis Blondell

Thesis Committee Approval:

Dr. Robert Voeks, Chair Dr. John Carroll, Department of Geography Dr. Jonathan Taylor, Department of Geography

Summer, 2016

ABSTRACT

Plants have long served as a primary source of medicines for humans. In the mid-

1990s, that began to change. Innovations in genomic biotechnology appeared on the scene. Some researchers felt frustrated by the recently adopted Convention on Biological

Diversity. Bioprospectors looked elsewhere for an additional source of medicines and found it in microbial organisms. Bioprospecting had entered a new era.

How and why this expansion into microbial bioprospecting occurred, and what it means for the future of bioprospecting, is the subject of this thesis. Viewed historically, certain events since the European Age of Discovery and onward, are responsible for driving bioprospecting into this new era.

Microbial organisms of the greatest interest are called extremophiles. These organisms thrive in places long thought impossible for life. Microbial bioprospecting plays out in distinctly different geographies than that of plants. Humid tropical rainforests have given way to fumaroles in America’s Yellowstone National Park. Deep sea vents thousands of feet beneath the ocean surface offer opportunities for bioprospecting. The frozen expanse of Antarctica has become the focus of bioprospectors. Searching for microbes in two of these three locations has not only changed how bioprospecting is conducted, it has the potential to create geopolitical tensions.

TABLE OF CONTENTS

ABSTRACT ...... i

LIST OF TABLES ...... vii

LIST OF FIGURES ...... viii

ACKNOWLEDGMENTS ...... x

Chapter 1. FROM PLANTS TO MICROBES ...... 1

2. BIRTH OF BIOPIRACY ...... 11

Seeds of a New Era ...... 11 Chapter Introduction ...... 11 Bioprospecting as Biopiracy ...... 11 The Theft of Cinchona ...... 14 Quinine’s Final Chapter ...... 20 Chapter Summary ...... 21

3. BIOPROSPECTING: A MORE HUMANE AGE ...... 22

Bioprospecting as Science ...... 22 Chapter Introduction ...... 22 Bioprospecting Enters the Laboratory ...... 23 The Role and Wonder of Secondary Compounds ...... 25 Ethnobotany and Ethnopharmacology: A Linkage is Formed ...... 30 Ethnobotanical Knowledge in Early Drug Development ...... 30 Chapter Summary ...... 33

4. THE AGE OF SYNTHESIZED MEDICINES ...... 35

Bioprospecting in the 1900s ...... 35 Chapter Introduction ...... 35 Bioprospecting Tries to Detach Itself from Plants ...... 35 The First Mass Screening Program ...... 38 The Continuing Linkage between Ethnobotany and Drug Development ...... 41 Ethnobotany Goes Psychedelic ...... 41

iii

Ethnopharmacology: Another Link in the Chain ...... 42 Chapter Summary ...... 44

5. TWO DECADES THAT CHANGED BIOPROSPECTING FOREVER ...... 45

Bioprospecting from 1980-2000 ...... 45 Chapter Introduction ...... 45 Farnsworth’s List ...... 46 Return to Paradise ...... 47 The Long March to the Convention on Biological Diversity ...... 49 Movements to Protect Indigenous People and Traditional Knowledge ...... 50 Chapter Summary ...... 54

6. TRADITIONAL KNOWLEDGE ...... 56

A Key Driver of Bioprospecting’s New Direction ...... 56 Chapter Introduction ...... 56 Traditional Ethnobotanical Knowledge: Who Owns It? ...... 57 Candidates for Biopiracy: Three Case Studies ...... 63 Quassi the Genius or Quassi the Biopirate? ...... 63 The Myth of the Rosy Periwinkle ...... 65 Salvation from the South Pacific: Kava ...... 69 Chapter Summary ...... 73

7. THE CBD AND IMPACTS ON ETHNOBOTANICAL RESEARCH………... 75

The CBD in Practice and the Nagoya Protocol ...... 75 Chapter Introduction ...... 75 The Convention on Biological Diversity: A New Era, New Problems ...... 76 Ethnobotanical Research post-CBD: Hard Lessons in Chiapas ...... 77 The CBD in the New Century: A Work in Progress ...... 85 Impact of the CBD and the Biopiracy Narrative ...... 85 The Nagoya Protocol: Bandage or Cure? ...... 86 Bioprospecting in the Digital Age ...... 87 Chapter Summary ...... 88

8. THE PHARMACEUTICAL INDUSTRY ...... 90

From Rainforest to Pharmaceutical: The Long, Uncertain, and Expensive Road ...... 90 Chapter Introduction ...... 90 Green Gold Reborn ...... 91 The Myth of (Easy) Green Gold ...... 95 The Value of Traditional Ethnobotanical Knowledge in Drug Discovery: Map or Maze? ...... 100 Pharmaceutical Companies as the Good Guys: Two Case Examples ...... 107

The INBio-Merck Benefit Sharing Agreement: A Revolutionary Pact ...... 108 Shaman Pharmaceuticals: A Painful Reality ...... 109 Chapter Summary ...... 112

9. BIOPROSPECTING IN AFRICA ...... 113

An Argument for Contemporary Biopiracy ...... 113 Hoodia: A Case Study ...... 113 Hoodia v. the Rosy Periwinkle ...... 118 Chapter Summary ...... 119

10. A NEW ERA IN BIOPROSPECTING ...... 120

Microbial Bioprospecting ...... 120 Chapter Introduction ...... 120 Microbial Organisms: Endless Possibilities ...... 120 The Extreme Worlds of Extremophiles ...... 123 Types of Extremophiles ...... 126 Chapter Summary ...... 131

11. MICROBIAL BIOPROSPECTING IN AMERICA’S NATIONAL PARKS ..... 133

A New Bioprospecting Frontier ...... 133 Chapter Introduction ...... 133 Bioprospecting for Hot Gold in the World’s First National Park ...... 134 Bioprospecting in America’s National Parks: An Activity Reborn ...... 140 Yellowstone and Beyond: Bioprospecting or Bio-destruction? ...... 145 Bioprospecting in America’s National Parks: Concerns and Issues ...... 146 Chapter Summary ...... 148

12. BIOPROSPECTING IN THE BLUE ABYSS ...... 150

Finding Pharmaceutical Treasures at the Bottom of the Sea ...... 150 Chapter Introduction ...... 150 Bioprospecting for Blue Gold ...... 150 Marine Bioprospecting: The Cost Barrier ...... 158 Bioprospecting on the High Seas and the Ambiguities of UNCLOS ...... 161 Bioprospecting on the High Seas: No Guidance from the CBD ...... 166 Chapter Summary ...... 169

13. BIOPROSPECTING FOR COLD GOLD ...... 170

Bioprospecting in Antarctica ...... 170 Chapter Introduction ...... 170 Bioprospecting for Microbes at the Bottom of the World ...... 171 Bioprospecting in Antarctica and Geopolitical Tensions ...... 175

Bioprospecting in the Arctic ...... 183 Chapter Summary ...... 184

14. MICROBIAL BIOPROSPECTING: A NEW ERA ...... 186

Great Potential, Great Obstacles ...... 186 Review ...... 186 Lessons Learned and Concluding Thoughts ...... 189 Looking Forward ...... 191 What Will Become of Plants? ...... 191 The Future of the Future of Bioprospecting ...... 193

APPENDICES ...... 194

A. FARNSWORTH’S LIST OF 47 MEDICINES FROM TROPICAL PLANTS ...... 194 B. DRUGS DERIVED FROM TRADITIONAL ETHNOBOTANICAL KNOWLEDGE ...... 196 C. YELLOWSTONE NATIONAL PARK’S EXTREMOPHILE HABITATS...... 199 D. DEEP OCEAN RIDGES AND THE MECHANICS OF HYDROTHERMAL VENTS ...... 200 E. ANTARCTICA IS A NEW FRONTIER IN BIOPROSPECTING ...... 201 F. ACRONYMS AND ABBREVIATIONS ...... 202

REFERENCES ...... 203

LIST OF TABLES

Table Page

1. A Partial List of Pharmaceutical and Industrial Uses of Antibiotics Derived from Microbial Organisms ...... 124

2. Habitats of Extremophiles ...... 130

3. A Sampling of Current Pharmaceutical Products Derived from Marine Organisms ...... 157

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LIST OF FIGURES

Figure Page

1. A map of the Americas circa 1860 showing an image of the Cinchona spp. and depicting its general habitat in the Andes Mountains (green region) ...... 16

2. Discovery of important alkaloids from plants: 1800-1900 ...... 24

3. Tropical rainforests (unfrosted area) possess great biodiversity, but are restricted to a small band near the equator ...... 29

4. Taxus brevifolia twig and bark ...... 40

5. Long road to the Convention on Biological Diversity ...... 54

6. The former African slave Quassi, along with a map of Suriname, and his namesake plant ...... 64

7. Where in the world did TEK for the rosy periwinkle originate? ...... 69

8. The remote island chain of Vanuatu in Oceania became the focal point of biopiracy accusations regarding the possible theft of traditional ethnobotanical knowledge during the kava craze of the 1990s...... 70

9. Chiapas State in Mexico was the region where a legitimate research project was derailed by outside NGOs ...... 79

10. Drugs discovered from ethnobotanical leads ...... 104

11. Yellow boundary represents approximate extent of Hoodia gordonii ...... 114

12. The pool that started it all ...... 135

13. Yellowstone National Park as seen through visible light (left) and infrared (right)...... 139

14. Researchers from WHOI photographed the first hydrothermal vent known as a “black smoker” ever seen by humans ...... 154

viii

15. A white smoker ...... 154

16. A black smoker ...... 154

17. Exclusive Economic Zones (hashed areas) as demarcated by UNCLOS ...... 163

18. Boundaries of maritime sovereignty as demarcated by UNCLOS ...... 164

19. A comparison of the landmass of Antarctica and the United States ...... 171

ACKNOWLEDGMENTS

Little did I know that flipping through an ecological law journal in the CSUF

Library, followed by a brief question to Dr. Voeks, “What’s microbial bioprospecting?” would result in a thesis more than a year later.

A good number of people deserve credit for that. Firstly, Dr. Voeks, for sharing with me his voluminous knowledge of ethnobotany and all aspects therein. His attention to detail saved me from numerous pitfalls and served as an inspiration. Any shortcomings on the following pages are mine alone. Hope to see you again in Lençios, professor.

Much gratitude is reserved for Dr. John Carroll and Dr. Jonathan Taylor for coming on at the last minute to serve on the thesis committee. Dr. Carroll, thanks for everything else you did. Dr. Taylor, much appreciation for always making time for graduate students.

Special thanks to the fellow grads: J.C., Anay, Vaishali, Kai, Shelly, Cindy,

Cheryl, Skyler, and Donna. They always listened to my complaining. And I did a lot.

The following faculty deserve notice: Dr. Drayse, I will not forget that semester when I did double-duty in two of your courses; Dr. Wu, I do not think that there is anything about Remote Sensing that you do not know; Dr. Xu, your knowledge on population geography is endless; Dr. DeLyser, your “little yellow book” turned out to be quite useful, although it may not show; Dr. Zia Salim, for endless patience and advice on presentations and posters. Much thanks to Dr. James Miller for his inspiration and

devotion. Finally, thanks to Prof. Leaa Short, Dr. Juliet Zaidi, Prof. Vanessa Engstrom,

Prof. Brian McCabe, and Prof. Scott Williams.

CHAPTER 1

FROM PLANTS TO MICROBES

When the desiccated body of Ötzi the Iceman was discovered lodged among rocks and ice in the Italian Tyrolean Alps in 1991, the mystery of what caused Ötzi’s death and how he came to be at that particular location when he died around 3,300 BCE, consumed a great deal of interest. Less discussed but equally important was the botanical medicine carried by the Neolithic sojourner. The fungus Piptoporus betulinus, commonly known as the birch bracket, was intended as a laxative to purge the parasites in Ötzi’s intestines that researchers have determined afflicted him (Crocq 2007; Mateo, Nader, & Tamayo 2001;

Oeggl 2009). Ötzi’s organic medicine was not the oddity of a remote mountain Stone

Age tribe, but an integral part of humankind’s millennia-long co-evolution with medicines from plants and botanical material.

Five thousand years later, humankind’s enduring partnership with medicinal plants transitioned into a new era. When the National Park Service signed a benefit sharing agreement with Diversa Corporation in 1997, it marked a historic shift from plants and fungi as a source of medicines to an increasing emphasis on microbes.

How and why that expansion to microbial bioprospecting occurred, and what the future of microbial bioprospecting holds, is the subject of this thesis. It is the contention of this thesis that certain identifiable events during the past several hundred years inexorably drove bioprospecting into the new era of microbial bioprospecting, and that if

2 it had not been for these specific events occurring at select times from the European Age of Discovery onward, this movement towards microbial bioprospecting would not have taken place in the time frame that it occurred.

Thousands of years before Ötzi’s time humans accessed plants as a source for new medicines (Fabricant & Farnsworth 2001; Gurib-Fakim 2006). Medicinal uses of plants allowed humans to survive and thrive. In the present age, technology like automated bioassays (Miller 2011) furthered the search for new medicines from plants.

By the mid-1990s, advancements in molecular technology had improved the screening of microorganisms for various applications (Hunter-Cevera 1998). From this time forward microbial organisms would gain increasing importance in pharmaceutical bioprospecting.

Bioprospecting can involve investigating biodiversity for “genetic and biochemical resources” (Reid, Laird, Gamez, Sittenfield, & Gollin et al. 1993, 3).

Bioprospecting often utilizes the traditional medicinal knowledge of indigenous societies to identify potential pharmaceuticals (Davidov 2013). For strictly pharmaceutical applications, bioprospecting can be the isolation and screening of bioactive compounds in plants for drug development (Kumar & Tarui 2007). For the purpose of this thesis, bioprospecting can be thought of as the search for plants whose properties benefit humans for medicinal purposes.

Bioprospecting encompasses a broad canvas of issues and concerns related to developing commercial products from biological resources (Lucchi 2013). Due to this, definitions of bioprospecting can be extremely generalized, benign, specific, contentious, or political, depending on the desired effect. Bioprospecting in contemporary times is often equated with biopiracy. Shiva (2007) denounces bioprospecting as a “sophisticated

3 form of biopiracy” (308). Such contemporary claims of biopiracy contributed to bioprospecting’s shift toward microbial organisms by motivating researchers to look beyond plants for new sources of medicines.

Historical events have shaped bioprospecting over the past several hundred years.

In turn, acts of bioprospecting have shaped history. Bioprospecting during the European

Age of Discovery often involved harsh and unscrupulous acts perpetrated against indigenous peoples by their (largely) Spanish and Portuguese conquerors. The colonization of the Americas in the 1500s decimated native populations by exposing them to European diseases. It also unleashed syphilis and other tropical maladies upon

Europeans (Voeks in press).

It was believed that hidden within the mysterious tropical rainforests was a medicinal “green gold” that would provide cures for the afflicted. Many Europeans believed that the godless indigenous peoples possessed knowledge in the use of these plants that would save the good Christians of Europe from much suffering. A centuries- long desperate quest began to wrest this knowledge from indigenous populations by whatever means necessary. Incidences of murder, torture, bribery, and theft to obtain this knowledge were considered necessary acts of persuasion by colonizing powers (Voeks in press; Zupanov & Xavier 2014). These historical acts of biopiracy have shaped perceptions of bioprospecting in contemporary times.

Historical acts of biopiracy laid the foundation for some in the developing world to view contemporary bioprospecting as a new wave of colonialism (Shiva 2004). Public perception of bioprospecting became controversial (Brown 2003). Many indigenous peoples and other stakeholders came to view any form of bioprospecting as biopiracy

(Isaac & Kerr 2004). This rationale stigmatized legitimate bioprospecting and in some instances halted research (Belejack 2003). Bioprospectors looked elsewhere for potential sources of medicines and found it.

Now bioprospecting is shifting to the diverse world of microorganisms. This new era of bioprospecting focuses on a very specific set of microbial organisms: extremophiles. These are microbes that inhabit some of the most inhospitable places on the planet like thermal vents (fumaroles), familiar to most through the images of boiling hot springs in Yellowstone National Park. Green gold has become “hot gold.”

Nor is the bioprospecting of extremophiles restricted to land. Deep below the ocean’s surface at depths that would crush a human body, extremophiles have been found that are already offering commercial applications in a variety of sectors. These extremophiles not only withstand intense pressure, but alternately thrive in temperatures of several hundred degrees, or in acidic or saline water that would kill most organisms

(Synnes 2007). Microbial bioprospecting for “blue gold” in maritime waters has gained a good deal of interest in the past decade.

Then there are the extremophiles on Antarctica and in the waters off its coastline.

Potentially useful microbial organisms have been located in frozen ice sheets, dry desert valleys, and chilly Antarctic waters. Polar extremophiles have been located in the ice sheets on Greenland (Hughes, Pertierra, & Walton 2013). The bioprospecting “cold rush” for polar microbial organisms has begun.

This shift or expansion toward microbial bioprospecting is the sum of many events over a long temporal scale. Voeks (in press) and Zupanov and Xavier (2014) laid out how the quest for green gold during the European Age of Discovery led to biopiracy.

The importance of ethnopharmacology to bioprospecting and drug discovery has been well documented (Ghorbani, Naghibi, and Mosaddegh 2006; Heinrich and Gibbons

2001). Farnsworth and Soejarto (1985, 1989) reignited interest in plants as a source of medicines by evaluating their economic potential. This helped to cultivate a perception in the public’s mind of Amazonia as a medicine cabinet filled with wonder cures (Davidov

2013). A need to protect indigenous peoples’ traditional knowledge and provide opportunities for benefit sharing led to the promulgation of the Convention on Biological

Diversity, or CBD (Aguilar 2001; Asebey & Kempenaar 1995; Conklin 2002; Peterson

2001; Timmermans 2003). Berlin and Berlin (2003, 2004) and Brown (2003) critiqued how the CBD and a few environmental Non-Governmental Organizations (NGOs) worked to confound rather than facilitate legitimate research. Recently, microbial organisms known as extremophiles have garnered attention as a source for pharmaceutical applications (van den Burg 2003).

Separately, each event is beneficial for illuminating stages in bioprospecting’s history. Yet it creates the impression that each episode has no connection with preceding chapters in bioprospecting’s evolution. To understand why only a few years separate the adoption of the CBD and the ascendance of microbial bioprospecting, historical connections need to be observed. By merging transient developments of bioprospecting into a continuous narrative, it is possible to ascertain how and why the shift toward microbial bioprospecting occurred, and what it means for the future of the quest for medicinals from nature.

Most literature has ignored how contemporary claims of biopiracy contributed to a growing interest in microbial bioprospecting. Some arguments of contemporary

6 biopiracy are based on long-past incidents such as quinine from the cinchona tree

(Mgbeoji 2006). The rosy periwinkle is often cited as a modern case of biopiracy to demonstrate indigenous peoples are being excluded from economic riches (Reid 2010;

Timmermans 2003). A few have pointed out the complexities of such examples (Brown

2003; Harper 2005; Malik 2005; Osseo-Asare 2014). Almost no literature has connected claims of biological theft and acts of biopiracy to the transition from higher plants to microbial organisms.

Literature is equally scarce when it comes to providing a comprehensive assessment of the transition to microbial bioprospecting. Discussions of microbial bioprospecting in America’s national parks (Bryson & Kaczmarek 2000; Doremus 1999;

Wood 2000), in marine waters (Dionisi, Lozada, & Olivera 2012; Synnes 2007; Haefner

2003), and in Antarctica (Jabor-Green & Nicol 2003; Lohan and Johnston 2005; Puig-

Marco 2014), all take place in separate contexts. Virtually no literature links the search for new chemical entities in America’s national parks, the abyssal depths, and Antarctica in a unified theme that addresses the movement toward microbial bioprospecting.

Linking past claims of biopiracy with the new era of microbial bioprospecting will require examining key developments in bioprospecting over a several hundred-year period.

This thesis is divided into fourteen chapters. The first chapter serves as an introduction. Chapter two examines an actual act of biopiracy during the European Age of Discovery. This and similar events planted seeds of animosity and distrust that would frustrate and ultimately thwart contemporary ethnobotanical researchers and bioprospectors. These contemporary allegations of biopiracy created a developed-

7 developing world divide that was a fundamental component in shifting bioprospecting toward microbes.

Chapter three surveys the transformation of bioprospecting in the 1800s as scientific inquiry began to assert itself over the domain of medicinal plant treatments.

Previously, bioprospecting had focused on plant simples (a leaf, root, etc.) for medicinal uses. Little thought was given to what made the plant useful. This changed in the late

1700s as scientific inquiry isolated bioactive agents within plants (Cordell 2000). This chapter makes the case that by the end of the 1800s, the lines between ethnobotany, ethnopharmacology, and bioprospecting were starting to merge. Further, the close association between ethnopharmacological research and ethnobotanical research positioned these two fields as one in the minds of indigenous peoples in developing countries. Equating ethnobotanical researchers as agents of pharmaceutical companies would have profound influence on the growing interest in microbial organisms.

Chapter four follows the merging of bioprospecting with modern technology. By the 1930s, synthesized drugs derived from plant compounds began to gain widespread use (Naranjo 1995). In the 1950s, the National Cancer Institute began an industrialized approach to bioprospecting (Frisvold & Day-Rubenstein 2008). This was the first time a methodical effort using traditional medicinal uses of plants was conducted with cutting- edge technology. The outcome of this mass screening and its implications on later research will be discussed.

Chapter five covers only twenty years from 1980 to 2000, but arguably the most important in bioprospecting’s history. Two important points will be discussed: a belief that tropical rainforests represented reservoirs of green gold and the adoption of the

Convention on Biological Diversity (CBD). Both will be shown to be key in moving bioprospecting toward an emphasis on microbial bioprospecting.

Chapter six addresses how misperceptions of traditional ethnobotanical knowledge played a key role in generating interest in microbial bioprospecting. As portrayed in the generic biopiracy narrative, a pharmaceutical company identifies a plant used as a medicine from the traditional medicinal knowledge of a specific group. Yet plants are not fixed within discrete boundaries. Neither are groups of people and their knowledge. Sorting out which group originated what discrete item of traditional knowledge becomes byzantine. This chapter shows that establishing who originated traditional knowledge and how that knowledge, if any, played into a specific drug development, is far more complex than the accepted narrative.

This leads to chapter seven where the impact of the CBD on ethnobotanical research is examined. The CBD was heralded as a solution that would prevent the theft of indigenous medicinal knowledge and allow indigenous peoples to profit from that knowledge. It will be shown the CBD erected barriers to research that it was supposed to facilitate. In many instances indigenous peoples were negatively impacted. Non-

Governmental Organizations (NGOs) became major actors in the bioprospecting narrative. This chapter shows that at least one NGO furthered its own political agenda rather than voicing concerns of indigenous groups.

Chapter eight examines the process of drug development based on bioactive compounds from plants. Contemporary narratives of biopiracy are based on a relatively uncomplicated plant-to-drug development process. By following the rise and failure of the much-hyped Shaman Pharmaceuticals this chapter counters that narrative. It will be

9 shown that drug development based on traditional ethnobotanical knowledge is an uncertain road to producing a successful pharmaceutical.

The rosy periwinkle is often cited as a prime example of biopiracy during the past century. Yet the African continent holds a much stronger argument for contemporary biopiracy. Chapter nine presents a case study of Hoodia that constitutes one of the most unambiguous acts of recent biopiracy. By contrasting Hoodia with the rosy periwinkle this chapter helps clarify the differences between real and imagined biopiracy.

Chapter 10 introduces bioprospecting’s next era. It discusses why microbes called extremophiles are of such great interest. Chapter 11 focuses on the movement from the so-called green gold of plants to the hot gold of thermal features. Fumaroles, or vents of super-heated water, can be found on land or deep offshore. Many of them are found in the

United States. America’s national parks are now the focus of microbial bioprospectors.

The National Park System is now promoting microbial bioprospecting revenue sharing agreements. How this came to be and what it means for the future of America’s national parks will be discussed.

Chapter 12 goes underwater to the abyssal depths of the ocean to investigate another sector of microbial bioprospecting. Thousands of feet beneath the ocean surface, where no light reaches and the pressure is dozens of times that at the surface, extremophile microbes have been found that offer many pharmaceutical and industrial applications. Just as perplexing as how microbes can exist in such bizarre and harsh environments, is the confusion surrounding who owns or who can access these microbes.

This section explores the question of whether the United Nations Convention on the Law

10 of the Seas is prepared for the political disputes that will certainly arise as countries compete to find the next miracle microbe on the ocean floor.

Chapter 13 concludes discussions on the third frontier of the expansion to microbial bioprospecting. This chapter ventures to the frigid climate of Antarctica.

Prohibited from national ownership but claimed by many states, Antarctica has become a focus of microbial bioprospecting. How these resources will be managed on the third largest continent is unclear. This leaves open the possibility for international conflict as countries scramble to grab their seat in the next bioprospecting gold rush.

Chapter 14 concludes with a review and analysis of what has been learned, plus insights that can be applied as bioprospecting enters the microbial era. A brief discussion on the future of bioprospecting concludes the thesis.

CHAPTER 2

BIRTH OF BIOPIRACY

Seeds of a New Era

Chapter Introduction

To understand how contemporary claims of biopiracy have shaped modern bioprospecting and thus pushed bioprospecting into its new era of microbial extremophiles, it is necessary to revisit a prior act of biopiracy. This instance is the theft of cinchona seeds by the British during the 1800s. This act of biopiracy is instructive for several reasons. It is a well-documented case of biopiracy; it was committed by a colonial power against indigenous peoples and specifically Bolivia; and the theft of cinchona seeds is used in present times to bolster claims of biopiracy. In the following chapters, we will learn how the ramifications of this act rippled through history to impact the future of bioprospecting.

Bioprospecting as Biopiracy

The arrival of Europeans in the Americas unleashed a bewildering number of diseases upon indigenous peoples. Thousands of years of geographic isolation left the native populations vulnerable to diseases from Europe and Africa. In certain places like the Caribbean island of Hispaniola, catastrophic depopulation from disease occurred within decades. Nor were Europeans immune to the germ exchange. Syphilis began

12 visiting death upon European populations not long after Christopher Columbus returned from his initial voyage to the Americas (Voeks in press).

Europeans unwittingly delivered upon themselves one of the most virulent diseases they encountered during colonial expansion into the American tropics.

Ironically, the need to possess an antidote to this disease was responsible for an act of biopiracy that rippled down to present times.

Malaria was established as far north as southeastern England by the 1600s. It is believed that both European settlers and enslaved Africans brought two species of the protozoan parasite (Plasmodium vivax and Plasmodium falciparum) to the Americas

(Gurib-Fakim 2006; Packard 2007). Malaria proved an effective buttress against the colonization of tropical regions in Africa and the Americas. Death rates for Europeans in

West Africa reached 483 per 1,000 during the 1700s. In the New World places like

Jamaica experienced a death toll of around 200 per thousand (Schiebinger 2004).

With European medicine relying largely upon poorly trained (if at all) surgeon- barbers, bleeders, herbalists, and astrologers, a desperate need arose to find cures for malaria and other illnesses. Since indigenous peoples in the Americas had been combatting diseases largely through medicinal plants, Europeans turned to these same plants for a cure (Voeks in press). One well known physician commented that in the

1700s, European medicine was so ineffectual that discovering new drugs was either a result of an accident or relying on the New World “savages” (Schiebinger 2004, 73).

Some of this desperation was rooted in the Doctrine of the Signatures which held that God endowed specific plants with cures for specific diseases (Schultes & von Reis

1995). Since Adam and Eve, according to Christian beliefs, had been cast out of the

Garden of Eden and some considered the tropics to be the site of the Garden of Eden, there was a belief that tropical plants held the cure for many diseases. Pharmacopeias that accompanied Europeans to the New World did not survive, and alternatives had to be found (Voeks in press). It was not until the invention of the Wardian case (a small glass terrarium) that plants could be successfully transported across great distances (Wirten

2008). To find medicinal properties in tropical plants the Spanish, Portuguese, and other colonial powers during that period, turned to the very people who were being destroyed by colonialism: the indigenous populations of the Americas.

Understandably, the indigenous people were not keen on divulging medicinal plant knowledge to those in the process of enslaving them. To entice the locals Europeans used tactics that ranged from gifts to threats of violence to torture (Voeks in press). Still, the degree of success in acquiring medicinal knowledge that proved an effective weapon against diseases is in question. What indigenous people viewed as theft was seen as bioprospecting by colonial powers. The unwillingness of indigenous people to divulge medicinal plant knowledge was viewed by colonial powers as secretive resistance to obstruct the spread of Christianity (Zupanov & Xavier 2014).

The Portuguese were the originators of “green imperialism” in the Colonial Age.

Economically important plants were cultivated in a number of their colonial possessions.

Plants served a practical importance other than economic wealth. Medicinally useful plants provided a natural pharmacopoeia for engenhos (sugar plantations), slaves, and casados (settlers) (Zupanov & Xavier 2014).

The Jesuit order amassed an impressive pharmacopeia to assist their missionary endeavors. The medicinal plant knowledge of the indigenous peoples was assiduously

14 recorded and experimented with by the Jesuits. These drogas do sertao, or drugs of the hinterland, also proved effective not only in healing the indigenous “pagans” but also the

Portuguese (Zupanov & Xavier 2014, 528). The Jesuits were such good observers of the local pharmacopoeia that they gained a near monopoly on one medicinal plant that would save lives in both Europe and the tropics, and led to one of the first acts of actual biopiracy (Voeks in press; Zupanov & Xavier 2014).

The Theft of Cinchona

One of the biggest coups of either duplicity or ingenuity in securing medicinal plant knowledge that returned actual economic benefits (as well as saving millions of lives) was bark from the cinchona tree. Theft of cinchona seeds from Andean countries is one of the few well-documented instances of a medicinal plant (not an industrial plant like rubber) being stolen from indigenous peoples by a concerted, well-funded, and persistent state-sponsored initiative.

The cinchona tree of concern here is one of over two dozen species in the genus

Cinchona, which is part of the Rubiaceae family to which the coffee plant belongs. Carl

Linnaeus, the famous Swedish taxonomist, supposedly named the tree (spelling the name wrong in the process) after a countess whose malaria was treated by the tree’s bark. More commonly the bark is referred to as Jesuit’s bark, Jesuit’s powder, Peruvian bark, or fever bark. It is now known that an alkaloid in the tree’s bark called quinine is responsible for its remarkable ability to combat Plasmodium falciparum, the parasite that causes most of the fatalities from malaria (Balick & Cox 1996; Gurib-Fakim 2006; Voeks in press).

The cinchona bark’s miraculous powers first came to the attention of Jesuit priests not long after Spain’s conquest of the Andean region of Peru, Bolivia, Ecuador, and

Colombia. Quinine is an often-cited example of biopiracy, due to documentation from the first experiments of cinchona bark by Jesuit priests, to the concerted efforts of colonial powers to cultivate the tree for themselves. That documentation can be traced back to

1653 in writings by Gaspar Caldera de Heredia, a Spanish physician. Indigenous peoples who were being led to Quito to work in mines had to ford a mountain stream. The frigid waters induced chills, which the local Indians treated by stripping cinchona trees of bark, pounding the bark into a powder, and mixing it with warm water. Jesuits accompanying the Indians noticed that the drink dramatically reduced shivering soon after its ingestion.

The Jesuit priests tested the potency of the concoction on malaria victims, thinking it might have some effect on the intense shivers induced by the disease. The drink proved surprisingly effective. Not only did it reduce the shivers of malaria victims, it appeared to cure the disease. Exports of the “fever bark” or “Jesuit’s bark” became a major economic export for Spain to the Old World (Voeks in press, 13-14; Zupanov & Xavier 2014, 530).

Although Robert Talbor gained notoriety in 1670 by curing royalty in Britain and

France with his concoction of cinchona bark, the source of the bark remained elusive

(Kaufman & Ruveda 2005). A Frenchman named Nicholas de Blegny wrote in 1682 that the cinchona tree was an “Indian” tree, but it was unclear if “Indian” referred to the subcontinent or people in the New World (Voeks in press, 32). Not even a botanist’s sketch existed of the tree which was actually endemic to the high, remote Andes

Mountains (Balick & Cox 1996). Eventually, collective minds deduced the tree’s natural habitat was somewhere high in the Andes (see Figure 1).

This was backed up in 1737 by William Arrot, a physician from Scotland who was practicing medicine in Peru. He is credited with the first field reports of the tree in its

16 native habitat (Voeks in press). Efforts moved to identifying cinchona’s curative powers.

Attempts in the laboratory to synthesize quinine’s bioactive compounds failed. Joseph

Pelletier and Joseph Caventou, both French chemists, isolated the alkaloid compound found in cinchona bark in 1820, but they had no luck with synthesizing a drug (Balick &

Cox 1996; Kaufman & Ruveda 2005). More forceful measures were needed.

Figure 1. A map of the Americas circa 1860 showing an image of the Cinchona spp. and depicting its general habitat in the Andes Mountains (green region). Map from Lazaridis, 1860, Library of Congress, Geography and Map Division, Washington, D.C., G32901860.L3; cinchona image from Roth & Streller, 2013; habitat adapted from Honigsbaum, 2003.

Colonial narrative at the time was reluctant to admit that the indigenous peoples had been using cinchona bark as a medicinal remedy for centuries. The imposed

European narrative was that an intellectually and culturally backward people could not produce a medicine more effective than what European minds could conceive. For a long time it was accepted that the Spanish introduced the healing wonders of the cinchona bark to the Andean peoples (Voeks in press). It was upon this Euro-centric belief that a centuries-long initiative to steal cinchona seeds was constructed.

Unlike other miracle cures of the time, Jesuit’s bark had lasting powers. The

Andean countries found that by luck of nature they had a much desired product in their possession. Colombia exported over six million pounds of cinchona bark in 1880. The

Andean countries of Bolivia, Colombia, Ecuador, and Peru attempted to maintain this monopoly by banning exportation of cinchona seeds or seedlings (Balick & Cox 1996).

To colonial powers like Britain, it seemed unfair that European people were held hostage by the monopoly of quinine by culturally backward countries. This control had to be broken. This belief led to the narrative that those South American countries within whose borders the cinchona tree naturally grew could not be trusted with managing the world’s sole supply of an effective antidote to malaria. A good deal of this manufactured hysteria rested upon how the bark was harvested. Andean cascarilleros would fell the tree to strip it of its bark, thereby killing the plant. Yet the tree would soon grow back, more vigorous than ever. This method of harvesting, while at first shocking, ensured a constant supply of cinchona bark. However, this minor point was over looked by British authorities (Voeks in press). Some, like Sir Clements Markham, later president of the

Royal Geographical Society, insisted New World peoples should surrender cinchona as a

18 repayment for all the good Europeans had done, such as providing them with a variety of agricultural crops and fruits, to livestock, including horses (Wirten 2008).

Britain was eager to secure seeds of the cinchona tree and establish its own plantations as means of extending its colonial hegemony in India and West Africa. In both places malaria had taken a horrifying toll on British lives. Sir William Jackson

Hooker, director of the prestigious Kew Gardens which was a repository for plants sampled (or pirated, depending on your view) from all over the world, was one of the main principals in the conspiracy. A plan was developed to be conducted jointly by the

Kew Gardens and Britain’s Secretary of State for India (currently under British control).

It called on whatever means necessary to collect and smuggle cinchona seeds out of their native habitat in direct violation of the export ban. Once in Britain’s possession they would be planted in India. Anticipated rewards would be British lives saved, reduced costs from exporting millions of dollars in cinchona bark, and British control over malarial-infested regions in Africa and India (Voeks in press).

Letters reveal geographer Clements Markham (the same Markham previously mentioned) and botanist Richard Spruce, were to secretly use “bribes and threats” to obtain and smuggle cinchona seeds to India (Voeks in press, 36). The plot included purchasing land high in the Andes where collections could be stockpiled. Richard Spruce eventually collected and smuggled over 100,000 seeds and 637 seedlings of Cinchona succirubra back to India. Markham focused on collecting another species, Cinchona salisaya. Markham’s efforts ended when locals got wind of his activities and chased him out of the country. Although Markham was able to bring along cinchona seeds during his escape, they perished by the time he reached India. Still, enough of Spruce’s seeds made

19 it back to Sri Lanka (Ceylon) to allow the British to produce enough quinine to maintain the health of their British colonists (Kaufman & Ruveda 2005; Voeks in press).

Attempts at stealing cinchona seeds persisted. After all, there was a vast sum of money waiting for those willing to engage in the necessary skullduggery. Charles Ledger was one such opportunist. Although not officially sponsored by the British government,

Ledger knew that anyone willing to provide British officials with cinchona seedlings would be handsomely rewarded. Earlier in the 1840s, Ledger had taken upon himself to obtain cinchona seedlings. He employed a local Bolivian native named Manuel Incra

Mamani who was especially adept at discriminating between different species of cinchona (Voeks in press). This was important because not all cinchona species (there are

40) contain the same level of malaria-resistant quinine alkaloids. The seedlings earlier obtained by Spruce and sent to India had a lower level of quinine than other species

(Balick & Cox 1996).

Despite his initial failure to obtain cinchona seedlings with high quinine levels,

Ledger moved back into the biopiracy business in 1865 when Mamani re-contacted him.

This time Mamani had several bags of Cinchona ledgeriana, the species with a high level of quinine alkaloids. Ledger paid Mamani $500 with instructions to obtain more. Ledger returned to England to sell the seeds (Balick & Cox 1996; Voeks in press). However, due to an earlier attempt by Ledger to sell low-quality quinine seedlings to the British government, and that a suitable plantation of cinchona trees (although with low quinine levels) was currently growing in India, Ledger’s offer was refused. Dismayed, Ledger sold the seedlings to a Dutch government official for $20. The Dutch sent the seeds to their colony on Java. Within a few years the Dutch discovered the $20 investment had

20 secured them a species with 13 percent quinine alkaloid levels—the highest ever recorded. Within half a century the Dutch controlled 97 percent of the global quinine market (Balick & Cox 1996).

For the Dutch, this was a long-awaited reward. An earlier effort by Justus

Hasskarl, head of a botanical garden on Java, duplicitously secured a load of cinchona seeds in 1852, only to find they had low quinine levels. Ironically, Cinchona ledgeriana

(formerly called Cinchona salisaya) was the very species obtained by Markham but whose seeds died enroute to Britain (Wirten 2008).

And what of Mamani, the Indian whose keen intellectual abilities and persistence had finally paid off in securing so many seedlings of high quality quinine that ultimately saved millions of lives? After selling the seedlings to Ledger, Mamani was arrested and tortured by Bolivian police, and died shortly after (Balick & Cox 1996; Voeks in press).

Quinine’s Final Chapter

The quinine saga did not end there. In the 1940s, the world was again seized by quinine hysteria. The Dutch had stockpiled nearly their entire supply of cinchona bark in

The Netherlands. As the Nazis occupied The Netherlands and the Japanese Imperial

Army occupied Dutch cinchona plantations in Indonesia, Allied forces were faced with having no antidote to malaria for their forces fighting in the Pacific and Africa. Once again, Andean countries home to the cinchona species found themselves crawling with foreigners desperately seeking cinchona. The Americans employed small armies of local people to strip cinchona trees of bark. Millions of pounds of cinchona bark were shipped back to America for pharmaceutical companies like Merck to attempt to find a synthetic alternative. Ironically, it was two Nazi agents who came to the Allies’ rescue. They

21 smuggled a considerable amount of pure quinine out of Amsterdam and sold it to an

American representative in South America. This allowed the Americans to produce the necessary quinine needed for the war effort. However, a structurally complex chemical quinine was not totally synthetically reproduced until 1944 (Balick & Cox 1996;

Kaufman & Ruveda 2005).

Chapter Summary

The discussion on the theft of cinchona seedlings is relevant to this thesis for several reasons. First, the unsanctioned removal of cinchona seeds in blatant violation of an export ban represented a clear and premeditated act of biopiracy. Second, it was largely state-sponsored. Third, it robbed countries that were home to the cinchona tree of a valuable endemic export. Fourth, unlike contemporary claims of biopiracy, it was well documented. Finally, it had far reaching public health outcomes.

The worldwide availability of quinine made possible by this act of biopiracy saved millions of lives. But it was also an act of blatant biopiracy. It is this act of past biological theft, more than any other that created lasting animosity and suspicion among indigenous peoples and enhanced the North-South dualism. To this day, it is a prominent example of biopiracy to which denouncers of bioprospecting point. This act of biopiracy rippled through the decades and set the stage for what would come over a hundred years later: the promulgation of the Convention on Biological Diversity. That came in the same span of time that bioprospecting expanded into microbial organisms.

CHAPTER 3

BIOPROSPECTING: A MORE HUMANE AGE

Bioprospecting as Science

Chapter Introduction

This chapter follows bioprospecting as it transitions from a blinded Cyclops to a more precise scientific endeavor. It ends by demonstrating how the emergence of two new scientific disciplines became forever linked. This linkage had profound ramifications on shaping the future direction of bioprospecting in the next century.

Previously, any plant that hinted at holding a possible cure ignited a scramble to obtain it. During the 1800s, this slowly changed. Scientific inquiry started to replace superstition. This began with identifying secondary compounds in the laboratory.

By the end of the 19th century ethnobotany had become a recognized scientific field. Laboratory analysis relied heavily on ethnobotanical field research. This focused research more precisely on targeting plants used by indigenous people for medicinal uses.

This had two future impacts. First, it forever linked ethnobotanical field research with laboratory efforts to develop drugs. Secondly, through this linkage with drug development, ethnobotanists became a target for lingering bitterness over earlier biopiracy like quinine. A field that initially dedicated much of its time to the innocuous cataloging of plants would, by the end of the 1900s, be accused of participating in biopiracy.

Bioprospecting Enters the Laboratory

Bioprospecting by colonial powers largely fulfilled two parallel goals. One was to obtain economically commercial plants such as rubber and spices. The second was to discover medicinal plants that could supplement medical practices that had changed little in the past several hundred years (Voeks 2004; Voeks in press).

During the 1800s, plant pharmacopeias still reigned. By some accounts, plants composed 90 percent of all medicines as late as 1930. Yet a subtle shift was taking place in bioprospecting. The 1700s saw science merge with medicine in a substantial way. That carried through into the 1800s as chemistry slowly began to unlock the mysteries of secondary compounds (Naranjo 1995). By the early 20th century, drugs like aspirin had been synthesized (Rishton 2008). Scientific inquiry was shaping bioprospecting into a precise endeavor.

This was largely due to the development of a science called pharmacognosy.

Pharmacognosy was coined in 1815 to describe the general study of drugs derived from plants or animals (Gurib-Fakim 2006). Scientists lost no time in doing so. Morphine, a secondary compound in the opium poppy, was isolated in 1806 (Rishton 2008). Quinine was isolated in 1820 (Fellows & Scofield 1995). Like dominoes falling, the 1800s saw alkaloid after alkaloid isolated from plants (see Figure 2). With these successes, pharmacognosy influenced all future pharmaceutical disciplines (Gurib-Fakim 2006).

This includes one not recognized as an official field until the end of the next century: ethnopharmacology.

In 1803, French ethnobotanist Louis Theodore Leschenault de la Tour led the first successful ethnopharmacological research. He observed indigenous peoples on the island

24 of Java prepare a poison using ingredients from a plant that was then applied to arrows.

Leschenault secured the primary plant used in the preparation of the poison and submitted the samples to other scientists. They determined the poison acted on the spinal cord to induce asphyxia and death. About 10 years later, Pelletier and Caventou identified strychnine as the bioactive alkaloid. It is one of the first known instances of a field researcher documenting the use of a plant by an indigenous group, then extending the research into the laboratory (Holmstedt & Bruhn 1995).

Figure 2. Discovery of important alkaloids from plants: 1800-1900. The isolation of alkaloids (secondary compounds) from plants progressed during the 1800s. The investigation of bioactive compounds in plants adopted a growing reliance on ethnobotanists in the field. By the end of the century, early pharmacognosy and ethnobotany were starting to overlap. Accompanying arrow indicates a generalized timeline of ethnobotanical influence on drug research. Graphic created by author; dates adapted from Raviña (2011) and Sneader (2000).

In Africa, poisoned arrows derived from the plant Strophanthus kombe were the object of another pharmacological investigation. Dr. John Kirk accompanied David

Livingstone on his infamous journey to Zambezi between 1858 through 1864. After accidentally getting some poison on his toothbrush that resulted in a much reduced pulse,

Kirk sent the plant to England for further study (Holmstedt 1995). By 1865, pharmacologists had isolated an alkaloid from the kombe vine. By 1885, the Burroughs

Wellcome & Company was marketing Strophanthin, a cardiac medicine with properties similar to digitalis (Hokkanen 2012).

Across the Atlantic in South America, curare, an infamous poison whose antidote had been the focus of intense bioprospecting by the continent’s colonial masters, helped move ethnopharmacology to the forefront. The fast-acting poison was used on the arrows of various tribes of indigenous peoples in the Amazon (Ghorbani, Naghibi, & Mosaddegh

2006). Curare-laced poison arrows had impeded colonization by the Spanish and

Portuguese. It had fascinated explorers and scientists like Alexander von Humboldt.

Claude Bernard, a French physiologist, was the first to carry out detailed laboratory analysis of the poison. He found the poison resulted in muscular paralysis (Heinrich &

Gibbons 2001). But Bernard was not satisfied. He emphasized the necessity of isolating the bioactive compounds from the foreign compounds with which the poison was mixed by the makers (Bernard 1966, cited in Heinrich & Gibbons 2001). That was no easy task.

In 1947, researchers finally isolated the bioactive compound in a particular type of curare made from Chondrodendron tomentosum (Heinrich & Gibbons 2001).

Through their meticulous scientific inquiry utilizing the microscope rather than brute force, Bernard and others had created ethnopharmacology—a direct result of bioprospecting, and a close relative of the then unnamed discipline of ethnobotany. Yet it was not until 1967, about 60 years after ethnobotany became a recognized discipline, that ethnopharmacology became a recognized field (Ghorbani, Naghibi, & Mosaddegh 2006).

The Role and Wonder of Secondary Compounds

Paracelsus, the noted alchemist, was by 1524 urging the scientific community to look beyond a plant like rhubarb that functioned as a purgative, and instead find the

26 bioactive agent in rhubarb that gave it a purgative effect (Cordell 2000). By the 1700s, the scientific community was questioning what agent in a plant was responsible for its effective medicinal properties. By 1780, Scheele was attempting to identify these properties in plants by investigating their organic acids (Sneader 1985, cited in Cordell

2000). Due to the advancements in scientific technology during the 1800s, it finally became possible to isolate bioactive compounds in plants. Building upon the research of

Scheele, by the beginning of the 1900s numerous bioactive alkaloids had been identified.

These included atropine, papaverine and codeine (Cordell, 1982; Sneader, 1985 cited in

Cordell, 2000). Science was slowly unlocking the medicinal secrets of secondary compounds.

Secondary compounds are basically quinine or aspirin for plants. That is, they function to protect the plant or allow it to thrive in a specific environment (Croteau,

Kutchan, & Lewis 2000; Seigler & Price 1976; Wink 1988). Unlike plants, humans are mobile. When we are in an unsuitable environment, or pursued by predators, we simply pick up and leave. Plants cannot do that. They are sessile. To survive in what might otherwise be an unsuitable habitat, or to ward off potential predators, plants use chemistry. These organic compounds form metabolites, known as secondary compounds.

Plant metabolism is broken into primary and secondary metabolism. Primary metabolism directs fundamental systems of plants such photosynthesis and respiration, along with manufacturing proteins for growth (Croteau, Kutchan, & Lewis 2000). Unlike secondary compounds, the purpose of primary compounds is quite clear (Seigler 1977; Swain 1977).

Secondary compounds baffled scientists for a long time. Plants have limited energy and it was puzzling why energy would be directed at forming metabolites that

27 have no apparent value to the plant’s growth (Seigler 1977; Seigler & Price 1976; Swain

1977; Wink 1988). It is now believed secondary compounds serve a variety of important functions, including defending the plant by producing chemicals that drive away potential predators; allowing the plant to adapt to certain environmental niches; assisting in seed dispersal by producing aromatic scents or colors that lure seed-dispersing animals; making the leaf structure resistant to pathogens; playing vital roles in relationships between the plant and its ecological environment; and serving as chemicals that assist in inter-species competition (Croteau, Kutchan, & Lewis 2000; Fellows & Scofield 1995;

Seigler 1977; Seigler & Price 1976; Swain 1977; Wink 1988). Secondary compounds should not be thought of as a bullet-proof vest for plants. Many secondary compounds may have a marginal affect, but over thousands of years this can substantially tilt the odds in the plant’s evolutionary favor (Swain 1977).

The appearance of eukaryotes (nucleated cells with complex functions) ~1.7 billion years ago opened the door for the evolution of secondary compounds. Flowering plants, known as angiosperms, possess the highest and most complex secondary compounds. Angiosperms evolved later than other plants, first appearing 138 to 63 million years ago during the Cretaceous Period. They have since come to dominate the plant kingdom. A burst of plant diversity began during the Eocene Epoch (starting ~55 million years ago) as plants shifted from abiotic dispersal to methods relying on animals

(Fellows & Scofield 1995).

Secondary compounds are derived from shikimic acids, amino acids, and acetate.

In turn, these starting materials are the basis for the three main groups of secondary compounds: the phenolics, nitrogen compounds, and terpenoids. The phenolics include

28 flavonoids, tannins, and quinones. Terpenoids include the terpenes, steroids, and carotenoids. They often regulate growth. Tannins are known for being effective antibiotics and antioxidants. Flavonoids assist in UV absorption. They play a fundamental role in reproduction by enticing pollinators and seed dispersers by adding color to fruits and flowers. Nitrogen compounds include most of the alkaloids. Over

12,000 have been isolated since the beginning of the 19th century. Alkaloids serve as a chemical defense against predators. They often have a bitter taste to the human senses.

Alkaloids can be toxic in the form of cocaine, nicotine, strychnine, or even caffeine

(Croteau, Kutchan, & Lewis, 2000; Fellows & Scofield 1995; Gurib-Fakim 2006; Swain,

1977).

How secondary metabolites relate to the new era in bioprospecting is due to their connection with biodiversity and where that biodiversity resides. Tropical rainforests are notoriously diverse. These regions of great biodiversity are situated between the Tropic of Cancer (23.5º N) and the Tropic of Capricorn (23.5º S). See Figure 3. A tropical rainforest experiences ~2,000 mm (79”) of rainfall annually and an average temperature above 24º C (75.2º F). In the Amazon basin precipitation can reach 3,000 mm (118”) annually. Earth’s surface is seven percent covered by tropical rainforests (Kricher 2011).

The physical attributes of tropical rainforest biomes result in a tremendous diversity of flora and fauna. Estimates have put species diversity for plants and animals between three million and 30 million. The Amazon may contain upwards of 80,000 different species of plants which amounts to 15 percent of all of the world’s plant species.

Biodiversity is concentrated in the tropics due to the latitudinal diversity gradient (LDG).

This means that as latitudes increase away from the Equator, be it north or south, species

Figure 3. Tropical rainforests (unfrosted area) possess great biodiversity, but are restricted to a small band near the equator. Adapted from Kricher, 2011.

diversity decreases dramatically. This is why a tropical rainforest can contain up to 250 or more tree species in a hectare (2.47 acres), compared to 10 to 15 per hectare in a temperate forest (Farnsworth & Soejarto 1991; Kricher 2011; Schultes 1994).

As species diversity increases, the number of secondary compounds (notably alkaloids) likewise increases. Tropical plants have been found to produce a higher number of alkaloids (Dannell & Bergstrom 2002; Rasmann & Agrawal 2011). Levin and

York (1978) found that tropical herbs have the highest alkaloid content when compared against shrubs and trees from tropical, sub-tropical, and temperate regions. Further, lowland tropical rainforests species contain the highest alkaloid percentage of rainforest plant communities.

Eighty percent of the world’s population is believed to rely on some degree of traditional medicinal healing (Gurib-Fakim 2006; Kim 2005; Moran, King, & Carlson

2001). Indigenous people make up around 4 percent of the world’s population (Rao

2006). Yet they comprise an estimated 95 percent of cultural diversity. Indigenous people typically reside in regions associated with a high degree of biodiversity (Firestone 2003).

Consider that with the known biodiversity of tropical regions and the plentiful alkaloids contained therein. Thus, it is understandable that if a researcher wishes to study traditional medicinal practices (or get insights into plants that may contain bioactive compounds), they head to a tropical rainforest. This is why rainforests and indigenous people are of great interest to bioprospectors.

Ethnobotany and Ethnopharmacology: A Linkage is Formed

As analysis of plant compounds advanced in the laboratory so did that of plant usage in the field by researchers. By the late 1800s, ethnobotany was coming into its own as a discipline. Pedanius Dioscorides probably became the first ethnobotanist of sorts when he published Materia Medica circa 77 CE. By the Renaissance Period, botany and human-plant relationships had moved from the domain of monks in the monastery to that of artists, and then to explorers and scientists in the field. In 1542, Leonhart Fuchs published a 400-page compendium of sketches showing plants native to Germany. In the

1700s, Alexander von Humboldt and his study (or creation) of biogeography inspired more curiosity about the plant world (Davis 1995).

Ethnobotanical Knowledge in Early Drug Development

In 1895, botanist John Harshberger at the University of Pennsylvania first used the term ethnobotany to define a discipline (Davis 1995). Harshberger saw ethnobotany as a subfield of phytogeography, or plant geography (Morgan 1995). Like bioprospecting, ethnobotany’s definition varies. Ghorbani, Naghibi, & Mosaddegh (2006) offer a sutiable generic definition for ethnobotany as the study of how people use plants for whatever

31 purpose. For this thesis, it is how people use plants for medicinal purposes. Thus, as a field of methodical research solidified around this new science, bioprospecting entered a new era. The harsh tactics discussed in chapter two were replaced by academic rigor.

Recall that pharmacognosy was meant to encompass the general study of drugs derived from plants or animals (Gurib-Fakim 2006). Since its inception in the early

1800s, pharmacognosy has seen its definition evolve. Balunas and Kinghorn (2005) wrote that in present times pharmacognosy encompasses “a broad study of natural products from various sources including plants, bacteria, fungi, and marine organisms”

(432). Further, the pair noted that the field melds many disciplines such as ethnobotany and ethnopharmacology into an “interdisciplinary science” (432). The American Society of Pharmacognosy acknowledges this wide scope. It defines pharmacognosy as “the study of . . . potential drugs or drug substances of natural origin, as well as the search for new drugs from natural sources” (Balunas & Kinghorn 2005, 432).

The last part in particular appears to merge with the goals of ethnopharmacology.

Holmstedt (1995) wrote that ethnopharmacology is generally considered “ . . . the interdisciplinary scientific exploration of biologically active agents traditionally employed or observed by man” (321). Lopez (2011) noted that ethnopharmacology draws on multidisciplinary research in the study of indigenous drugs, and that in many instances ethnobotany and ethnopharmacology have shared goals. Remember that Leschenault was

“an ethnobotanist” who launched the first successful ethnopharmacological investigation that linked field work and laboratory analysis (Holmstedt 1995, 321).

Both ethnobotany and ethnopharmacology are considered interdisciplinary fields

(Gurib-Fakim 2006). It is acknowledged that ethnobotany, including the study of

32 medicinally useful plants, is a multi-disciplinary endeavor drawing upon various methodologies (Davis 1995; Holmstedt & Bruhn 1995; Heinrich & Gibbons 2001; Lipp

1995; Morgan 1995).

Ethnobotany is an umbrella term that covers how humans use plants for foods, medicines, or most any other use (Farnsworth 1994). While ethnobotany does not primarily concern itself with developing pharmaceuticals from indigenous medicinal uses of plants, observing the pharmacological properties of a plant can form a component of research (McClatchey, Mahady, Bennett, Shiels, & Savo 2009). Harshberger felt that ethnobotany was an effective method to learn “ . . . new uses of plants . . . ” that could be applied to present times (Morgan 1995, 251). Davis (1995) wrote that part of the ethnobotanical practice of finding and cataloging new plants is “ . . . incorporating their usefulness into modern society” (40). Even before then, in 1874, “aborginal botany” was coined by Stephen Powers to describe the various uses of plants, including medicinal applications (Davis 1995, 43).

From the 1700s through the end of the 1800s, explorers and field researchers sent back plants to be tested, accompanied with exotic tales of the medicinal practices of indigenous peoples (Davis 1995). Ethnobotanists came to play a critical role in a pharmacological research. Hoffman (1995) noted this relationship when he wrote, “ . . . it was, and still is, the ethnobotanist who is the mediator between healers or shamans and medicinal chemists . . . ” (311).

Bioprospecting, generally regarded as “exploring biodiversity for new sources of natural products” (McClatchey, Mahady, Bennett, Shiels, & Savo 2009, 2) plays a role in

33 all three disciplines. As the 1800s drew to a close, a co-dependence developed between the early fields of ethnobotany, ethnopharmacognosy, and later on, ethnopharmacology.

This overlap was particularly apparent in one of the most well-known instances of drug development from a plant. For centuries various groups had used willow bark to treat pain from headaches and rheumatism. A richly documented ethnobotanical history of willow bark’s medicinal uses among indigenous peoples was the reason Felix Hoffman at the Bayer Company began experimenting with synthesizing salicylic acid, the bioactive compound in willow bark (Rishton 2008).

Medicinal plants were always of great interest to humans. Early written texts describing the various medicinal uses of plants carried forward into the modern age

(Schultes & von Reis 1995). With the burgeoning discipline of ethnobotany describing uses of medicinal plants (Hofmann 1995), and the new science of ethnopharmacology studying bioactive compounds in medicinal plants, the two fields became merged, if not in purpose, then at least in a symbiotic relationship. As the 1800s ended, the fields of ethnobotany and ethnopharmcology appeared to be on parallel tracks. Bioprospecting came to be seen as a useful instrument for both.

Chapter Summary

This interdisciplinary mingling is important in shaping the contemporary history of bioprospecting for two reasons. Scientific advancements in the 1800s, and the intellectual curiosity of what made plants medicinally potent, created the springboard for continuing research in the 1900s. Secondly, many indigenous people reside in developing countries, while the laboratories of ethnopharmacologists are in developed countries. This created a latent developed-developing world contrast that a hundred years later would

34 come back to haunt ethnobotanists who saw themselves simply as the contemporary generation of Harshberger and Schultes. Intentionally or not, bioprospecting became linked with drug development.

CHAPTER 4

THE AGE OF SYNTHESIZED MEDICINES

Bioprospecting in the 1900s

Chapter Introduction

This chapter follows bioprospecting through the technological advancements of the 1900s. In the 1930s, synthesized drugs from plants began to gain mass consumer appeal. In the 1950s, the National Cancer Institute (NCI) revolutionized pharmaceutical development with an industrialized approach to bioprospecting. Although not a huge success, it reignited corporate pharmaceutical interest in plant-based medicines.

By the 1970s, interest in plants as a source of medicines was starting to wane.

Biotechnology innovations in the form of mass-screening and bio-assays would eventually re-stimulate interest in plants as a source of medicines. Ethnopharmacology, so important during the previous century for placing bioprospecting on a scientific footing, became a recognized field. This set the stage for the 1980s which saw fundamental shifts in both the perception and the practice of bioprospecting.

Bioprospecting Tries to Detach Itself from Plants

By the late 1800s, medicine was shedding its image from a field dominated by quacks practicing bloodletting to a regulated and professional science. Still, medicine was greatly dependent upon plant-based medicines. At the start of the 1800s, at least 80 percent of medicines available were derived from plants (Makhubu 1998). Yet by the late

1800s and into the early 1900s, interest in medicinal uses from herbs was beginning to wane. The public had grown tired of “wonder cures” derived from herbs that were utterly useless. As a growing organic chemistry discipline promised new synthetic drugs, people began to turn away from plants as medicines (Schultes & von Reis 1995).

Although Merck & Company marketed morphine as the first synthetic drug in

1826, the sustained age of synthesized medicines started in 1899 when aspirin was developed by Bayer (Veeresham 2012). This was followed by barbital, another synthetic drug, about a decade later (Naranjo 1995). Synthetics for dentistry, including Novocain, were commercially available in America soon after 1891. At the end of the 1800s, and into the early 1900s, almost every year saw a synthetic drug released that legitimately treated a disease or relieved suffering (Tainter & Marcelli 1959).

By the 1930s, synthetic drugs appeared poised to revolutionize medical care. The development of drugs like dichlorodiphenyltrichloroethane (DDT), penicillin, and a new era of man-made polymers (Fellows & Scofield 1995) promised an age of unparalleled human health and comfort. The synthesis of sulphamides introduced the age of chemotherapy that same decade. Following World War II, medical drug technology increased to a point where, in the 1960s, 90 percent of all drugs were synthetic (Naranjo

1995).

Despite successes in the laboratory at synthesizing new drugs, renewed interest in bioprospecting and medicinal plants was reignited in the years leading up to and during

World War II. The global conflict saw huge amounts of money dumped into pharmaceutical research and development to create antibiotics and other drugs for the war effort (Petrova 2014).

The renewed interest in drugs derived from plants began when tubocurarine, a naturally occurring alkaloid in Chondrodendron tomentosum, was purified as D-

Tubocurarine in 1935 (Brown 2013; National Center for Biotechnology Information, n.d.). Tubocurarine had a long history of bioprospecting. This paralyzing neurotoxin used on poison arrows was first described by Peter Martyr D’Anghera in 1516 during travels in South America. Charles Waterton, a wealthy man acting as a caretaker for sugar plantations in Guyana, went bioprospecting for the legendary Wourali poison in 1812.

After two years he was successful. Efforts during the 1800s to use derivatives of the poison to treat muscle spasms caused by tetanus and strychnine poisoning were unsuccessful. Yet by 1942, the neuromuscular blocker was being used as a popular anesthetic (Brown 2013).

Penicillin, synthesized and mass-produced by the mid-1940s, was derived from fungi. This helped reignite interest in organic products as a source of antibiotics (Naranjo

1995; Rishton 2008). Another drug derivative from a plant began to be widely used for treating hypertension. Rustom Jal Vakil, an Indian cardiologist, extracted an alkaloid from Rauwolfia serpentine, known as the snake-root plant. Reserpine was soon synthesized and prescribed for combating hypertension (Gilani & Rahman 2005; Gupta

2002; Ishwarwal & Gupta 2006).

Other plant-derived drugs were gaining attention. Digitalis, reserpine, tubocurarine, cortisone, and the rosy periwinkle—medicinally useful extracts from plants—all had one thing in common: their medicinal uses were based in the folk medicine of “primitive” societies (Schultes & von Reis 1995, 287).

There were less pleasant reasons for the return to natural sources of drugs.

Concurrent to the development of potent pesticides like DDT and drugs like thalidomide, there was a growing realization these synthesized drugs could cause great harm (Fellows

& Scofield 1995). The scientific world once again returned to bioprospecting for medicinal plants, only this time armed with technology that was lacking only a few decades prior.

The First Mass Screening Program

In the 1958, the National Cancer Institute (NCI) initiated the Natural Products

Program. The purpose was to institute the world’s first mass-screening of natural products (plants) that might show bioactivity against cancerous tumors. The program was in conjunction with the United States Department of Agriculture (USDA) which had a robust program in collecting plants.

Thousands of plant samples were required. Collectors spread out over 60 countries, returning approximately 3,500 to 4,000 plants a year, of which about 2,600 came from the USDA. From the start of the program to the end of its initial phase in

1982, over 35,000 samples from plants and 114,000 extracts were analyzed for anti- cancer bioactivity (Aylward 1995; Cragg, Boyd, Cardellina, Newman, & Snader et al.,

1994). This encompassed 12,000 to 13,000 species of plants (Soejarto, Fong, Tan, Zhang,

& Ma et al. 2005).

During the approximately 20 years of the first screening program, about 9.7 percent of the plant samples and 4.3 percent of the plant extracts showed initial bioactivity. Out of those initial screenings, 2,000 isolated compounds were selected for additional screening. Just 17 of those merited clinical trials. From that 17 (Aylward

1995), just one compound (paclitaxel) went on to put the program in the headlines. Its trademark name was Taxol.

Taxus brevifolia is commonly known as the Pacific yew. Although the sample that ultimately yielded Taxol came from a tree in the Pacific Northwest, species of the yew genus grow in other parts of the world (see Figure 4). The T. brevifolia and T. canadensis species have been used in traditional medicines by North American indigenous cultures (Cragg et al. 1994). Still, the initial emphasis of the NCI screening program was not based on traditional medicinal knowledge, but rather on botanically related species that might possess similar bioactive agents. This was because the NCI felt

(at the time) using traditional medicinal knowledge had limited applications within the goals of the program. The NCI believed that since many tumors are internal and slow growing, societies lacking in advanced medical technology would have difficulty in detecting cancerous tumors; traditional medicinal practices used to treat external cancers would be poisonous if used internally; and that since cancer is largely a disease of extended age, tribal societies whose individuals had shorter life spans would be unlikely to experience internal cancerous diseases, and therefore possess less knowledge on that topic (Aylward 1995).

The NCI program was terminated in 1982 due to its failure to identify numerous anti-cancer bioactive agents. In the 1980s, interest in plant-based medicines continued to grow alongside innovations in screening technology. The NCI restarted its program from

1986 to 2004. This time collection focused on plants from tropical regions of the world which the NCI hoped would prove more successful in isolating useful bioactive compounds, due to the greater biodiversity and hence the greater number of secondary

Figure 4. Taxus brevifolia twig and bark. The drug Taxol derived from the bark was the result of the world’s first modern mass-screening of secondary compounds in plants. From Seiler, Jensen, Niemiera, & Peterson, 2015.

compounds that were expected to be found. Both botanical diversity and traditional medicinal treatment were considered as springboards to investigations (Aylward 1995;

Cragg et al. 1994; Soejarto et al. 2005).

Taxol’s journey from bioactive agent to marketable pharmaceutical is emblematic of the problems encountered in turning plant-based compounds into an effective drug.

The species of the tree that produces the highest quantity of paclitaxel is the Pacific yew.

But the tree is notoriously slow-growing. Producing one kilogram of Taxol requires upwards of 16,000 pounds of tree bark. Each tree yields only three to five pounds of bark and can require up to 100 years to produce mature bark. During the early stages of

41 development a stable supply of raw paclitaxel was questionable. The yew tree was in short supply, which made reliance on trees in the wild problematic. Concerns of over harvesting arose. The program looked stalled.

This created political concerns that after having invested millions in the research program, the NCI did not have the financial ability to bring the drug to market. In 1989, researchers at Florida State University developed a process to semi-synthesize paclitaxel.

Bristol-Myers Squibb licensed the process. In 1992, the FDA approved Taxol (the trademark name) as an anti-cancer drug. A year later Bristol-Myers Squibb released the drug on the market. Taxol was a huge success. By 2000, global sales had topped $1.5 billion (Frisvold & Day-Rubenstein 2008).

The initial phase of the NCI’s screening program did succeed in its purpose to locate an anti-cancer drug. But it is a sober reminder that out of the 114,000 plant-based compounds screened during 21 years of the initial screening program, only two extracts were considered major discoveries. One was paclitaxel, the other was camptothecin. Two other drugs were derived through semi-synthesis based on camptothecin (Cragg, Katz,

Newman, & Rosenthal 2012). Still, the NCI program proved that the mass screening of plants using modern technology could have pharmacological and economical value. This kept alive interest in bioprospecting for medicinal plants.

The Continuing Linkage Between Ethnobotany and Drug Development

Ethnobotany Goes Psychedelic

A second development served to attract the attention of both pharmaceutical companies and the public regarding traditionally-used medicinal biological products. In

1939 and 1940, two researchers published articles that later served to turn on a generation

42 to the wonders of natural hallucinogens. In 1939, American anthropologist Jean Bassett

Johnson published research on the use of teonanacatl mushrooms by the Mazatec people in the Mexican state of Oaxaca. A year later, ethnobotanist Richard Evans Schultes published an article on the same mushrooms that were once used as a hallucinogen by the

Aztecs (Heinrich & Gibbons 2001; Hofmann 1995; Johnson 1939; Schultes 1940).

Inspired by this research, Robert Gordon Wasson, an amateur mycologist, and his wife Valentina Pavlovna, traveled to Mexico to experience first-hand the power of these hallucinogenic mushrooms. Between 1953 and 1956, Wasson visited Huautla de Jimenez to witness the sacred velada ritual. Those experiences, published in Life magazine in

1957 as “Seeking the Magic Mushroom” generated wide public interest in hallucinogenic natural products (Heinrich & Gibbons 2001; Hofmann 1995; Schultes & von Reis 1995).

This came just as the social revolution of the 1960s was getting underway. The burgeoning psychedelic era of the 1960s saw people experimenting with hallucinogenic fungi for less than scientific reasons (Schultes & von Reis 1995). Nonetheless, it served to situate the medicinal possibilities of natural products in the public mind. This would become important two decades later.

Ethnopharmacology: Another Link in the Chain

It was during the 1960s that a third development acted to further link bioprospecting and pharmaceutical companies and forever change the course of bioprospecting. In 1967, the term ethnopharmacology was coined (Ghorbani, Naghibi, &

Mosaddegh 2006; Heinrich & Gibbons 2001). In 1979, the first issue of the Journal of

Ethnopharmacology was published (Soejarto et al. 2005). More than a hundred years after bioprospecting took its first steps into scientific analysis, ethnopharmacology

43 became a recognized and respected discipline. By the end of the century drug development from plants had moved from accidental discoveries and haphazard research to methodical screening and development (Petrova 2014).

This was the result of overlapping and merging in research objectives between ethnopharmacology and ethnobotany. Ethnobotany and ethnopharmacology both draw on field research to study traditional or indigenous medicinal knowledge (Soejarto et al.

2005). Bioprospecting and ethnopharmacological research are not always discrete activities (Heinrich & Gibbons 2001). Bioprospecting can be conducted by those inside or outside of a particularly scientific discipline (Soejarto et al. 2005). Further, ethnobotanical knowledge of a particular community or region can be valuable for bioprospectors investigating antidotes to infectious diseases (Heinrich & Gibbons 2001).

In some instances, ethnobotanists have formed close alliances with drug companies to discover leads for new pharmaceuticals (Berlin & Berlin 1994).

The previous chapter established the overlapping methods and objectives of ethnobotany, pharmacognosy, and ethnopharmacology, and the reliance on bioprospecting that they all share. Some regard bioprospecting as multinational companies tapping the traditional medicinal knowledge of indigenous groups (Warner

2006) for the purpose of developing new pharmaceuticals (Heinrich & Gibbons 2001).

Since ethnobotanical research often focuses on the traditional use of medicinal plants, it is challenging to parse the lines between ehtnopharmacology, ethnobotany, and bioprospecting. Ethnobotany’s multi-faceted connection to pharmaceutical bioprospecting was about to hurt academic researchers in a substantial way.

Chapter Summary

Significant developments occurred during the first eighty years of the 1900s that helped facilitate the eventual expansion toward microbial bioprospecting. A turning away from plants as a source of medicines in the early part of the century was partially due to a public tired of bogus medical cures. As more synthetic drugs hit the market, partially spurred by World War II, there was confidence that synthetic chemistry would provide copious amounts of new medicines.

A second notable event was the commencement of a decades-long screening program by the NCI. However, the first phase of the program produced just one major clinically approved drug. Still, hallucinogenic mushrooms kept alive public and scientific interest in natural products as drugs.

A third important outcome was the growing association between ethnobotany and ethnopharmacology, and their uses of bioprospecting. The unintentional linkage forged at the end of the 1800s between ethnopharmacology, bioprospecting, and ethnobotany was reinforced. This perceived close alliance with transnational pharmaceutical companies in developed countries would taint ethnobotany over the next two decades. Starting roughly in 1980, the events, crimes, and ghosts of the past several hundred years of bioprospecting would come to a denouement. Bioprospecting would change irrevocably.

CHAPTER FIVE

TWO DECADES THAT CHANGED BIOPROSPECTING FOREVER

Bioprospecting from 1980-2000

Chapter Introduction

This chapter explores a 20-year period that forever changed the course of bioprospecting. It did so by shaping fundamental perceptions of the rainforest, biodiversity, and holders of traditional knowledge. The outcome of the decades between

1980 and 2000 was a result of events that had unfolded over the past several hundred years of bioprospecting.

It was during this period that the term biopiracy was coined to reflect the growing distrust toward bioprospecting. Global media and Non-Governmental Organizations became major actors in shaping the biopiracy narrative. Public perceptions regarding the rainforests began to change. Indigenous peoples came to be seen as holders of medicinal knowledge that could provide remedies to many developed world afflictions.

Concerns over intellectual theft, rainforest destruction, threats to biodiversity, along with the simmering animosity over acts of biopiracy committed during colonial times, gave rise to the promulgation of the Convention on Biological Diversity (CBD). At the time the CBD was regarded as a significant development in protecting the intellectual property of indigenous peoples and ensuring their economic welfare. How effective the

CBD was in resolving these concerns will be addressed in a following chapter.

Farnsworth’s List

In 1985, Norman R. Farnsworth and Djaja Doel Soejarto published an article in

Economic Botany that had a significant impact on the direction of bioprospecting.

Farnsworth and Soejarto’s question was simple: if a plant became extinct what was the economic loss? The answer helped to rejuvenate interest as plants as a source of pharmaceuticals.

Farnsworth and Soejarto first looked at the number of plant-based pharmaceuticals. To qualify as plant-based, a drug could not have been manufactured without its basic bioactive agent coming from a plant. The pair found that roughly 23 percent to 25 percent of pharmaceutical prescriptions filled during the period 1959 to

1973 qualified as plant-based pharmaceuticals (see Appendix A for Farnsworth’s list).

Basing their studies on over-the-counter sales of pharmaceuticals derived from plants,

Farnsworth and Soejarto extrapolated the sales of plant-based pharmaceuticals in 1980 to be $8.112 billion. By their count at the time, they found that plants served as the basis for

40 drugs. Dividing those 40 plant-based drugs into $8.112 billion returned an estimated economic loss of a single species to be $203 million. Further, Farnsworth and Soejarto found that since 40 pharmaceuticals had been derived from 5,000 flowering plant species worldwide, it meant for that every 125 plants analyzed for bioactivity, one would become a marketable drug (Farnsworth 1993). With a possible upward limit of 422,000 plant species worldwide (Pitman & Jorgensen 2002), and tropical countries home to an estimated 125,000 of those species (Gurib-Fakim 2006), people took notice.

Return to Paradise

On Farnsworth’s list of medicines derived from plants were vinblastine and vincristine, both effective anti-cancer agents derived from the rosy periwinkle

(Catharansus roseus). This sparked a great deal of attention. The thought went, if medicinal plants were worth such a great amount, and Eli Lilly was raking in an estimated $100 million a year from vinblastine and vincristine (Frisvold & Day-

Rubenstein 2008), didn’t tropical countries that were home to these plants deserve to receive royalties? Green gold fever took hold in a number of tropical countries.

Concurrent to this was a growing assumption among the public that cures for numerous diseases could be found in a pristine tropical rainforest. This belief, produced by the developed world in periodicals, books, and movies like Medicine Man, constructed an untouched tropical paradise that was endangered by evil loggers or money-grubbing

Big Pharma (Aylward 1995; Davidov 2013; Voeks 2004; Voeks in press). However, indigenous peoples had carved out a millennia-long existence in the rainforest by intentionally altering the landscape, such as cutting and burning (Miller 2007; Voeks

2004). Along with providing for agricultural uses, such activities created areas where medicinal plants could grow, called disturbance pharmacopoeias (Voeks 2004).

Still, during the 1980s, the Amazonian rainforest was portrayed as the “lungs of the world” which helped construct the belief that the rainforest was the property of wealthy, developed countries (Goldman 1998, cited in Davidov 2013, 249). This environmental narrative of a disappearing Amazon became a “fantasy Amazon”

(Davidov 2013, 245).

In 1989, TIME magazine had a cover issue with an image of Earth as “Planet of the Year” (TIME Magazine 1989). The caption “Endangered Earth” with the Amazon visible on the front cover fed the perception of an environmental Armageddon.

Projections of impending destruction for the world’s plants, particularly those in tropical countries, were plentiful. Estimates of a 44 percent loss to tropical forests between 1940 and 1980 were circulated. Annual deforestation losses reached an estimated 75,000 km2. Projections of tropical forests disappearing in 30 years with an estimated 60,000 plants becoming extinct within half a century were made (Hamann

1991). Others estimated 27 percent to 33 percent of all known and unknown plant species fell under the status of threatened (Joppa, Roberts, & Primm 2011). Some even predicted a “goodbye, rainforests” scenario of complete devastation (Janzen 1987, cited in

Farnsworth & Soejarto 1991, 34). Nor were forests and plants the only victims. In

December of 1988, the Brazilian rubber tapper Chico Mendes was murdered (Hecht

1989). This was widely publicized and followed by the 1994 movie depicting his life, The

Burning Season.

Possibly, one of the most influential factors contributing to the changing perception of the rainforest, folk medicine, and bioprospecting in the mind of the public was the commercial success of Mark J. Plotkin’s 1993 book, Tales of a Shaman's

Apprentice: An Ethnobotanist Searches for New Medicines in the Amazon Rain Forest.

The book’s casual but compelling storytelling style generated wide public interest in indigenous medicines of Amazonian people. Although Plotkin’s goal was to create a written compendium of folk medicine for his host village in Suriname, the book helped

49 reinforce a perception that wonder drugs existed undiscovered in the rainforest, and that any deforestation could have profound consequences on the world at large.

Feeding into this was the growing demand for phytomedicines, also known as herbals and botanicals (a.k.a. dietary supplements). These products saw a huge growth in sales during this period. Sales reached an estimated $12.4 billion by 1993 (Gilani &

Rahman 2005; Mateo, Nader &Tamayo 2001), as those in developed countries sought the

Fountain of Youth on local pharmacy shelves. All of the above drove a sense of urgency.

Public perceptions of bioprospecting soured within two decades. When the

Journal of Ethnopharmacology published its first issue in 1979, bioprospecting was regarded as relatively benign due to the positive effects on both local and global communities. Also, there was the accepted belief that traditional (indigenous) medicinal knowledge was the common heritage of humanity (Soejarto et al. 2005). By the early

1990s, “biopiracy” had been coined to reflect the growing frustrations over the exploitation of indigenous peoples, justified or imagined (Hamilton 2006a; Robinson

2010). By the end of the 1990s, the field of ethnobotany, which included researchers like

Richard Evans Schultes—who had spent years living among indigenous peoples of the

Amazon region and documenting their traditional medicinal practices—had become a symbol of intellectual theft to some biopiracy critics (Brown 2003).

The Long March to the Convention on Biological Diversity

Despite the fanfare accompanying the adoption of the CBD in 1992, concerns about developing countries poaching biological material and traditional knowledge did not arise out of a vacuum. Changes in bioprospecting were related to slowly changing attitudes about indigenous societies.

Movements to Protect Indigenous People and Traditional Knowledge

The first major step was arguably the 1971 Barbados Conference sponsored by the World Council of Churches. Ostensibly organized to inventory the cultural and environmental plight of indigenous peoples in South America, the conference served to launch and promote an international movement for the rights of all indigenous peoples.

The “Declaration of Barbados for the Liberation of the Indians” became an internationally recognized document that established guidelines for researchers, religious groups, and governmental administrators interacting with indigenous peoples in the

Amazon region (Berlin & Berlin 2004; Brysk 1996).

Concerns of biodiversity acted as a bridge to the plight of indigenous peoples. In

1987, a push was made by the United Nations Environmental Program Governing

Council to formalize various and fractured treaties governing biological diversity on a global scale (Mgbeoji 2006).

Years before the Convention on Biological Diversity and the Earth Summit, there was a growing consensus that indigenous peoples and developing countries had the right to both protect and receive compensation for traditional knowledge (Cordell 2000).

The 1970s saw efforts to increase awareness of the medicinal value of plants in

Mexico. Mexican President Luis Echeverria instituted a program to encourage campesinos (peasant farmers) to supply raw plant materials for medicines, as part of his program to encourage pharmaceutical development of drugs from plants (Rose, Quave &

Islam 2012).

In 1976, the World Health Organization (WHO) passed Resolution WHA29.72 to bring attention to practitioners of traditional medicine. A year later, Resolution

WHA30.49 encouraged countries to use traditional medicinal knowledge. In 1978, the

WHO took it a step farther with Resolution WHA331.33. This asked countries to build an inventory of medicinal plants, assess scientific merit of the traditional medicinal uses, exchange information among groups and countries, and create centers for studying medicinal plants (Akerele 1991).

A decade later in 1987, resolutions passed at the Fortieth World Health Assembly urged member states to create programs to study, evaluate, cultivate, conserve medicinal plants, and ensure quality standards of drugs made from plants (Akerele 1991). From

1984 through 1987, the International Union for Conservation of Nature and Natural

Resources (IUCN) promoted the Plants Conservation Program to develop strategies for plant conservation and establish botanical gardens (Hamann 1991).

In 1990, Thomas Eisner from Cornell University fueled the growing interest in biodiversity conservation and benefit sharing. A year earlier in Goteborg, Sweden, the

International Society of Chemical Ecology adopted the Goteborg Resolution. This document called for using a small amount of profits from chemical bioprospecting for conservation programs in developing countries to counter the loss of biodiversity. Eisner pointed out that a loss of a species meant a loss of chemicals that could have profound medicinal applications. Eisner urged for chemical prospecting to be increased, particularly in the tropics, where potentially useful medicinal plants were facing extinction. He outlined a method where laboratories from developed countries would work in close association with tropical countries in screening plants. Under his plan, pharmaceutical companies would share profits with the host nations. Eisner proposed that initial funding come from debt-for-nature swaps that were gaining in popularity (Eisner

1990). Coming on the back of Farnsworth’s paper only a few years before, this helped feed the growing attention on the potential of traditional medicine.

The possibility of merging conservation with benefit sharing for indigenous groups was embraced by much of the international community. International Non-

Governmental Organizations (NGOs) set up organizations to assist indigenous peoples and make them economically self-sufficient. Profiting from their natural resources and traditional knowledge was the obvious option (Berlin & Berlin 2004).

Long before the CBD, a perceived lack of benefit sharing and overharvesting of resources was becoming an issue. According to Tedlock (2006) there were instances of outsiders “discovering” plants used by indigenous peoples without sharing any future profits from the plant’s uses (257). Others felt there was a problem with over-extraction of biological resources and a lack of inadequate benefit sharing in the years leading up to the adoption of the Convention on Biological Diversity (Takeshita 2001). Momentum was building to address these concerns.

The first meeting of the International Society of Ethnobiology (ISE) was organized in 1988 in Belem, Brazil. The conference members concluded that conservation programs should be established, indigenous peoples must be consulted in actions that affect them, and that compensation was deserved for the use of their knowledge and resources. While methods of prior informed consent (PIC) were not strictly addressed, no international scientific body had ever before recognized a need for guidelines in dealing with indigenous peoples (Berlin & Berlin 2004; Who We Are, n.d.).

In 1992, the same year that the CBD came into existence, the Charter of the

Indigenous Tribal Peoples of the Tropical Forests was published in Malaysia. It not only

53 recognized the contributions that traditional technologies of indigenous peoples make to the global society, but it demanded control over traditional forms of medicinal practices and knowledge (Moran, King, & Carlson 2001).

Rio de Janeiro, Brazil, was the ideal location to hold the 1992 United Nations

Conference on Environment and Development (UNCED). Brazil has a huge biodiversity in both flora and fauna, along with diverse cultural groups. The country contains around

25 percent of all plant species in the world (Verma 2002). Brazil has been described as a

“laboratory in situ” (Garcia & Ming 2012, 480). The UNCED, informally dubbed the

Earth Summit, was intended to focus on measures to protect countries like Brazil with a high biodiversity. For a timeline of events leading up the UNCED, see Figure 5.

After a little more than 10 days, the conference produced the Convention on

Biological Diversity (CBD) (Earth Summit 1997). In just 28 pages the modest document laid out guidelines for collecting plants and other biological resources; benefit sharing for the usage of these biological resources; obtaining prior informed consent; and acceptable conduct for field researchers (Neimark 2012; United Nations 1992). It could be said that, at the time, the CBD was regarded as a Declaration of Independence for plants

(biological products) and indigenous peoples.

The Convention on Biological Diversity had multiple objectives. It sought to conserve biological diversity, provide for the sustainable use of that diversity, and provide for equitable benefit sharing of that diversity. The CBD also sought to ensure proper legal access to biological resources while respecting the rights of those groups or countries that controlled such resources (Cragg, Katz, Newman, & Rosenthal 2012;

Schindel, Bubela, Rosenthal, Castle, du Plessis et al. 2015). A measurement of the CBD’s

Figure 5. Long road to the Convention on Biological Diversity. A timeline of developments leading to the promulgation of the Convention on Biological Diversity. Graphic by author. Collected from sources cited in text.

effectiveness on achieving its goals will be discussed later. But at that moment in 1992, the CBD was heralded as a breakthrough for indigenous peoples, researchers, and Big

Pharma alike. For better or for worse, bioprospecting was changed irrevocably.

Chapter Summary

This chapter established that although short in temporal scale, the twenty years from 1980-2000 were instrumental in shaping bioprospecting’s future.

A series of human rights declarations and conventions over the decades since

WWII eventually merged into a global effort to protect indigenous peoples. It culminated with the adoption of the CBD.

Concurrently, popular culture played a prominent role in changing the public’s mind from the rainforest as a forgotten wilderness, to the rainforest as the container of miracle drugs. As a result, bioprospecting began to be viewed unfavorably.

These events merged into a powerful force that changed bioprospecting irrevocably, but not necessarily how it was intended. Instead of acting to provide a framework where indigenous peoples would control and profit from indigenous knowledge, these two decades would ultimately be key drivers in moving bioprospecting toward microbes. How this unintended consequence came about will be discussed in the next two chapters.

CHAPTER SIX

TRADITIONAL KNOWLEDGE

A Key Driver of Bioprospecting’s New Direction

Chapter Introduction

One objective of the CBD was to protect the traditional ethnobotanical knowledge

(TEK) held by indigenous peoples by providing a structure for them to exercise control over their intellectual property. It was the alleged theft of TEK (also referred to as traditional medicinal knowledge (TMK)) that helped give rise to the now ubiquitous usage of the term biopiracy. But what is TEK/TMK? And who, if anyone, owns it? This chapter examines several instances where claims of biopiracy have been used to illustrate the theft of knowledge from traditional groups. Examples of the rosy periwinkle, kava, and an enslaved African named Quassi, demonstrate that delineating ownership of

TEK/TMK is more complex than what simplistic allegations of biopiracy would have us believe.

It is not easy to distinguish the origin of traditional ethnobotanical knowledge. As

TEK is assimilated by others through whatever form of contact, determining what groups deserve to participate in benefit sharing can be maddeningly frustrating. Which leads to the question, can any group actually own TEK? This chapter will demonstrate that ownership of TEK is highly complex, varies from case to case, and thus efforts to offer blanket remedies for such a complicated issue are ill-fated.

Traditional Ethnobotanical Knowledge: Who Owns It?

The history of humans is intertwined with plants. Thousands of years before recorded history humans most certainly were engaged in bioprospecting. Uses of medicinal plants stretching back 60,000 years to the Middle Paleolithic have been identified through fossils (Fabricant & Farnsworth 2001). Plants were traded extensively in the ancient world (Merson 2000). One of the earliest recorded expeditions to obtain plants was launched by Queen Hatshepsut of Egypt in 1495 BCE. She sent her emissaries to the Land of Punt in East Africa to obtain samples of the Boswellia tree. The tree’s aromatic resin was the source of frankincense, an important and lucrative incense of that period (Robinson 2010; Wynberg & Laird 2007). Oil derived from the Boswellia carteri tree contains boswellic acid, a compound with medicinal properties including anti-tumor activity (Frank, Yang, Osban, Azzarello, & Saban et al. 2009). In the Americas, the

Aztecs were known to have sent out expeditions to discover medicinal plants. The

Chinese assimilated plants from Southeast Asia (Lipp 1995).

India’s use of plants for medicinal and therapeutic purposes goes back to the

Rigveda over 4,000 years ago. The Pen Tsao, a compilation of 366 medicinal herbs, was published in 2500 BCE by the Chinese herbalist Shen Nung. By 1550 BCE, the

Egyptians had compiled over 700 medicinal remedies known as the Ebers Papyrus. The

Greek physician Pedanius Dioscorides authored a botanic compendium of over 600 species of plants called De Materia Medica. The book, published circa 77 CE, was so comprehensive in its description of medicinal uses for plants that it served as an authoritative source for almost 2,000 years (Garrity & Hunter-Cervera 1999; Gurib-

Fakim 2006; Jain 1994; Raviña 2011; Schultes & von Reis 1995).

War in ancient times played a role in dispersing medicinal knowledge of plants.

The Greeks adopted Yunani (Unani) medicinal practices around 400 BCE. This knowledge was adopted by Arab doctors and was later brought to India during the Mogul invasions (Jain 1994). Asian migrants ventured across the land bridge from Asia into the

Americas. The medicinal plant knowledge they possessed was modified as they acquired new medicinal knowledge (Raviña 2011).

Colonization in more recent times was a key factor in the exchange of medicinal practices. Colonizer and colonized assimilated each other’s knowledge of useful medicinal plants (Jain 1994; Morgan 1995; Osseo-Asare 2008; Robinson 2010; Schultes

& von Reis 1995; Wynberg & Laird 2007).

Absence of written documentation makes tracing TEK impossible. The type of plant used and the method of preparation or application were transmitted orally through the generations (Balunas & Kinghorn 2005). Complicating this is that the origin of a medicinal plant (genetic resource) and the traditional knowledge of its use may have originated in different locations (Dutfield 2013). Eventually, the precise geographic origins of specific plants were forgotten, if they ever were known (Robinson 2010). It is impossible to know if Ötzi’s people originated the medicinal knowledge of the fungus in his possession. Most probably not.

Contact between cultures and processes such as borrowing, appropriation, migration, and diffusion have been ubiquitous for so long that it is difficult to determine what is authentically indigenous. It is estimated 66 percent of the world’s population, and

75 percent to 90 percent of the population in developing countries, access 35,000 to

70,000 species of plants for some sort of medicine (Farnsworth & Soejarto 1991;

Hamann 1991; Schumacher 1991). It is logical to assume some of those uses and traditional knowledge may have been passed from and assimilated by different groups over time. Identifying a plant that grows in a number of countries but has TEK uses specific to one cultural group is problematic (Robinson 2010). Therefore, determining if the present holder of traditional knowledge is in fact the group that originated the knowledge, and if not, who has a right to compensation, is difficult at best (Chennells

2013).

Definitions of indigenous people are neither concise nor universal. The World

Intellectual Property Organization (WIPO) defines indigenous peoples (communities and nations) as “those which, having a historical continuity with ‘pre-invasion’ and pre- colonial societies that developed on their territories, consider themselves distinct from other sectors of the societies now prevailing in those countries, or parts of them.” Further, indigenous peoples are often distinguished by being creators, holders, and transmitters of traditional knowledge (World Intellectual Property Organization 2001, 23). Additionally,

“local” and “indigenous” communities can be interpreted as two distinct kinds of groups

(Chennells 2013).

Attempts to define traditional knowledge can be equally difficult. Quinn (2001) offers a satisfactory definition for discussion purposes here by defining it as “ . . . creation through a long period of time which has been passed down from generation to generation; new knowledge is integrated to the existing, as knowledge is improved . . .”

(293). Establishing the authors of this “improvement” can make it more difficult to decide the true ownership of TEK. There are other considerations. Chennells (2013)

60 points out that traditional knowledge may be interpreted to have a different meaning than that of indigenous knowledge.

Ownership of traditional knowledge is often viewed differently by those in the developed world and societies in developing countries. Customary rights began to clash with property laws when the United States passed the Plant Patent Act in 1930. This law allowed asexually reproduced plants to be patented. Prior to that, genetic resources, including plants, were generally viewed as a “common biological heritage.” Later, DNA techniques gave rise to genetically engineered plants (or biological resources) that could be patented (Merson 2000, 288). A landmark 1980 Supreme Court decision in Diamond v

Chakrabarty allowed the patenting of a genetically engineered bacterium developed to fight oil spills (Moran, King, & Carlson 2001; Tsioumanis, Mattas, and Tsioumanis

2003). These two developments created interest among the pharmaceutical and agricultural industries for medicinal plants used in traditional societies that could serve as the basis for new products (Merson 2000). A major goal of the CBD was to change the perception that plants were a “common heritage of mankind” and could be exploitated by anyone without regulation. The CBD was intended to give countries and indigenous societies control over biological resources within their domain (Ho 2006; Timmermans

2003).

For developed countries, the ability to patent biological material is key to intellectual property. According to the WIPO, a patent is “an exclusive right granted for an invention, being a product or process that offers a new technical solution to a problem.” To qualify for protection, the invention must be novel, meaning new and nonobvious; possess a new characteristic not found in prior art; must demonstrate an

61 inventive step; and be capable of industrial application or usefulness (World Intellectual

Property Organization 2001, 25).

A key component of contention between indigenous knowledge and patent laws rests in interpretation of what constitutes “prior art” (loosely defined as a previously existing work). U.S. patent law does not recognize prior art if it is not documented in writing (Finetti 2011; Ho 2006). This is problematic in indigenous cultures where such knowledge is transmitted orally through the generations, rather than through literary works (Sarma 1999). Some argue that claims of biopiracy such as the Indian Neem tree are more akin to patent disputes than outright intellectual theft (Chen 2006; Ho 2006).

Differences in beliefs leave the door open to claims of biopiracy.

Western view holds that patents serve to encourage investment and stimulate the market in a particular sector (Isaac & Kerr 2004). Granting patents for new pharmaceuticals is an example. Ho (2006) points out that indigenous people view traditional knowledge as part of their cultural composition and patenting any part of that knowledge is interpreted as an attempt to commodify their culture. Traditional knowledge is seen by indigenous peoples as shared by the entire community, whereas patents reward an individual. Traditional knowledge or the use of medicinal plants is embedded within a group’s culture, and often considered sacred (Ho 2006; Moran, King, & Carlson 2001).

Therefore, any economic incentive to profit from a patent may be morally objectionable

(Ho 2006). Collective or traditional knowledge can form the core of a people’s distinct cultural identity (Tsioumanis, Mattas, and Tsioumanis 2003). Stripping a people of their

TEK may be perceived as stripping them of their identify.

Holders of traditional knowledge may have substantially different veiws on what is patentable than people in developed countires. Posey (2002) points out that indigenous societies generally regard themselves as equal with nature, acting as its guardians and caretakers. Nature exists as an integrated extension of society. Whereas, in Western society technology, rather than nature, is regarded as an extension of the individual. Thus, many indigenous societies are concerned that pharmaceutical companies from developed countries will modify traditional knowledge of plants, and then patent the modifications through intellectual property laws (Timmermans 2003).

A significant barrier to determining ownership of traditional knowledge is its fluid nature. As in developed societies, traditional societies change as contributions from successive generations are assimilated (Timmermans 2003). Traditional knowledge is constantly being updated as a group adapts to changing environments (Finetti 2011; Ho

2006). Thus, traditional knowledge does not necessarily indicate ancient practices. It can be recently acquired (Finetti 2011; Ji 2011; Timmermans 2003). Traditional knowledge is generally regarded as possessing social significance. Traditional may also refer to the method in which the knowledge is transmitted to those within the group (Ji 2011). Yet without a clear record of contributions it is impossible to determine authorship.

This leads to the key component of the biopiracy debate, and by default, a key component of the transition to microbial bioprospecting: with what group or person does traditional knowledge reside? It is the overlapping transfer of this knowledge, willingly or forced, documented or undocumented, intra-group and inter-group, that creates such a messy problem in sorting out whose indigenous knowledge is whose.

To demonstrate this point, a survey of three disputes concerning ownership of traditional ethnobotanical knowledge will prove instructive. One deals with a person of which few have familiarity. The second is a popular drug often touted as a prime example of biopiracy. The third is a commercial drink derived from a root extract that rode the hype of tropical medicinal salvation. Each offers a unique viewpoint from which to parse the question pushing much of the biopiracy debate, and thus the future direction of bioprospecting: Who originated what knowledge?

Candidates for Biopiracy: Three Case Studies

Quassi the Genius or Quassi the Biopirate?

A likely instance of an individual assimilating TEK from others and using it for their own gain occurred in colonial Suriname during the 1700s. Ironically, it was not the

Dutch colonial masters co-opting the traditional knowledge of Amerindians, but an enslaved African who gained fame and riches for his miraculous grasp of local medicinal plants and practices.

Kwasimukamba was brought to Suriname in chains after being kidnapped in coastal Guinea in the early 1700s. Around 1730, Kwasimukamba, more commonly referred to as Quassi or Kwasi, “discovered” medicinal uses of a tree which quickly brought him great fame as a healer among enslaved people, Indians, and white colonists

(Price 1979). See Figure 6.

The bitter solution Quassi brewed from a tree helped fight fevers by lowering body temperature. The famous taxonomist Carl Linnaeus later named the tree Quassia amara in Quassi’s honor. The solution, known as Quassi Bitters, is not an official pharmaceutical, but remains available in Suriname. Quassi accumulated a great deal of

Figure 6. The former African slave Quassi, along with a map of Suriname, and his namesake plant. Quassi, known alternatively as Graman Quacy, Quassie van Timotibo, or Quasimukamba, likely biopirated traditional ethnobotanical knowledge from local tribes in Suriname dealing with the medicinal uses of Quassia amara (right). Quassi image from Voorheve & Field, 2007; map from ESRI 2014; photo from Sanchez in Medicinal Plant Conservation Newsletter, 2012.

wealth and notoriety for his medicinal skills. Not only did he come to own a plantation and slaves, he was showered with gifts from the Dutch government (Voeks in press).

Since Quassi was brought to Suriname as a child, he certainly did not acquire his acute medicinal skills in Africa. Rather, he must have assimilated his craft from the local indigenous population. Quassi’s Dutch masters assumed as much, lauding Quassi for

“ . . .the considerable knowledge which he was able to acquire from the Indians . . . ”

(Price 1979, 152).

Thus, Quassi was richly rewarded for medicinal knowledge he certainly did not originate. Was Quassi a biopirate for practicing these medicinal skills on the whites, even though any member of the local indigenous population could have done the same? How much of the indigenous knowledge did Quassi actually appropriate? No one knows.

Perhaps innate intelligence enabled Quassi to expand upon the practices of indigenous healers in a unique manner. Keep in mind that the Jesuits who experimented with

65 cinchona bark did not expect to cure malaria, they just intended to relieve the shivers caused by the disease. Or was Quassi just clever and astute in exploiting an opportune situation for his advantage? Is there a difference between that and biopiracy? Whatever the real answer, this forgotten bit of intellectual appropriation is probably a better case for biopiracy than the next example that is cited far more widely.

The Myth of the Rosy Periwinkle

Very few people would suspect that a roadside weed is the source for two potent anti-cancer drugs, but that is exactly the case for the rosy periwinkle. Also known as

Catharanthus roseus, this plant has sustained the biopiracy narrative and saved millions of lives for several decades. Of all the biopiracy allegations, this plant most perfectly fills the claims of Big Pharma exploiting the poor, indigenous healer. Of all the biopiracy claims, it is perhaps the shakiest.

Biopiracy proponents would have you believe this: for generations the rosy periwinkle, native to Madagascar, was used by indigenous healers for a variety of healing practices. Then a scientist from a developed country, upon hearing of this miraculous plant, absconded with it back to a pharmaceutical lab. Soon after—presto—drugs based on the plant cured thousands suffering from cancer and earned millions for Big Pharma.

And Madagascar got zilch. In this instance, fact is neatly tucked behind the curtain.

The geography of the rosy periwinkle underscores claims of intellectual theft. The number of periwinkle species found on Madagascar leads botanists to believe that the plant is endemic to that landmass. Seven species of Catharanthus are found on

Madagascar, with an eighth endemic to India (Osseo-Asare 2014). The plant figures into

66 the medicinal healing properties of cultures on six continents, including islands in the

Caribbean. The rosy periwinkle is now ubiquitous worldwide (Brown 2003).

The ubiquitous distribution of the species raises the question of how the plant was able to travel so extensively. During the period of Swahili trade, it is believed that the periwinkle made its way to Eastern Africa, Asia, South Asia, and from there the rest of the world. No written documentation exists pre-colonization on Madagascar until French subjugation during the 1600s. Early records indicate that Catharanthus was useful for heart ailments, headaches, or venereal disease. On Madagascar, the rosy periwinkle is thought of as a weed with many homeopathic applications. Sometimes the entire plant was utilized, or only leaves, or just roots. Malagasy fishermen chewed the periwinkle roots to relieve hunger and provide stimulation. Various medicinal uses also come from places as scattered as Jamaica and the Philippines (Osseo-Asare 2014).

Starting in the 1920s, a desire arose to find a less expensive drug to treat diabetes.

It is uncertain where using periwinkle leaves to treat diabetes originated. Possibly, it evolved from homemade recipes, or clinical investigations, or a mixture of both. South

Africa, where a recipe dating to the 1920s was found, is a prime location (Osseo-Asare

2014). An April 1926 issue of the Journal of the American Medical Association (JAMA) cites a study on the action of vinca on the blood sugar of rabbits in South Africa. In

January of 1929, JAMA carried an article on efforts in Australia to treat diabetes with

Vinca rosea, or the rosy periwinkle (Desai 2013; Epstein 1926; JAMA 1926; JAMA

1929). Still, the Caribbean figures prominently in the rosy periwinkle saga because it is the island of Jamaica, not Madagascar, from where the idea originated to test

Catharanthus for bioactivity (Harper 2005).

Both Filipinos and Jamaicans reportedly used Catharanthus to treat diabetes

(Harper 2005). Leaves from a rosy periwinkle in Jamaica made their way to a researcher in Canada in 1958. Soon the pharmaceutical giant Eli Lilly began testing the plant.

Screening for bioactivity against diabetes went nowhere, but the plant’s extracts were promising for bioactivity against tumors (Chen 2006). In 1965, Lilly received a patent for the process to produce the alkaloids known as Oncovin (vincristine) and Velban

(vinblastine) (Osseo-Asare 2014). The drugs were shockingly successful. Vinblastine has proven 80 percent effective in fighting Hodgkin’s disease while vincristine, in combination with other drugs, has demonstrated a 90 percent remission rate against leukemia. Total annual drug sales are reported to be US $160 to $180 million annually, although the patents have expired (Harper 2005; Newman 1994; Osseo-Asare 2014).

Somehow, Madagascar and not Jamaica became the focal point of the rosy periwinkle saga. Claims of biopiracy and lack of revenue sharing do not end with the manufacture of the drug. Eli Lilly could not synthesize the alkaloids and continued to need periwinkle plants to produce the medicine. Fifteen tons of periwinkle plants are required to produce one ounce of vincristine (Newman 1994). Eventually, cultivation operations were moved to Texas to ensure a stable supply (Brown 2003).

The large amount of periwinkle plants required to produce the drugs played into the belief that Eli Lilly was contributing to deforestation on Madagascar. Yet the rosy periwinkle, like many important medicinal plants, is not found in pristine rainforests, but rather in disturbed areas (Harper 2005; Newman 1994; Osseo-Asare 2014; Voeks 2004).

In some places like Florida the rosy periwinkle is regarded as a nuisance weed (Brown

2003). Yet that has not stopped the rosy periwinkle from being used as the poster child for rainforest devastation.

The rosy periwinkle issue raises a number of questions. If biopiracy occurred, who was the victim? It is hard to make a case for Madagascar due to the plant’s wide distribution over the centuries. As we have seen, early records indicate that the plant’s usage to treat diabetes possibly originated in South Africa. Yet no diabetes drug was ever derived from Catharanthus extracts. Since the rosy periwinkle was circulated widely among many cultures, with each no doubt adopting the previous culture’s knowledge and contributing their own inspiration to the mix, does any single group actually deserve credit? Or revenue sharing? One might say Jamaica deserves credit, but the plants used in

Eli Lilly’s initial research reportedly came from India (Brown 2003). Should India lay claim as well?

Nor is there a straight link from indigenous medicinal practices to commercial pharmaceutical, since the drugs ultimately developed from the rosy periwinkle were not used to fight diabetes. Since all of these cultures supposedly used the rosy periwinkle to treat diabetes, and not cancer, is it possible for any group to lay claim to ownership of traditional knowledge? Therefore, in this instance it is difficult to make a legitimate claim of biopiracy. See Figure 7.

In Quassi’s case, claims of biopiracy are uncertain. In the instance of the rosy periwinkle, they are dubious. Further, both happened centuries or decades ago. Yet contemporary claims regarding a popular plant from the South Pacific are no less clear in demarcating lines of biopiracy and theft of traditional ethnobotanical knowledge. This

Figure 7: Where in the world did TEK for the rosy periwinkle originate? Question marks (added by author) are all possible sites. World map from ESRI 2014; photo of rosy periwinkle from Sain & Sharma 2013.

next case demonstrates how claims of biopiracy and theft of intellectual knowledge can be directed by one indigenous group against another.

Salvation from the South Pacific: Kava

Most people associate the island chain of Vanuatu with James Michener’s Tales of the South Pacific. The 80-plus islands are located ~1500 miles northeast of Australia.

Until the 1980s, the islands were known as the New Hebrides, having received the name from Captain Cook who explored the island group in 1774 (see Figure 8). Johann Georg

Forster was struck by the intoxicating power of an endemic plant called kava, and named the tuberous plant Piper methysticum—basically, intoxicating pepper (Singh 1992).

Kava is used across Oceania for religious purposes and as a relaxant that encourages social geniality. In the 1990s, Vanuatu become the focal point for a kava craze. Largely pushed as a relaxant drink that relieved stress and anxiety, the kava root was also sold dried and ground into a medicinal powder. Kava was included in herbal

Figure 8. The remote island chain of Vanuatu in Oceania became the focal point of biopiracy accusations regarding the possible theft of traditional ethnobotanical knowledge during the kava craze of the 1990s. This time much of the dispute was between indigenous island groups in Oceania. Yellow oval indicates the approximately 80 islands that form Vanuatu. The yellow arrow indicates the island group’s relative location. Photo of kava plant (upper left) from Ligo, 2016. Basemap from ESRI, 2014.

supplements and homeopathic medicines. Some claimed it had applications in weight reduction and could help insomnia. Others believed ingestion induced a hypnotic state

(Lindstrom 2009).

As kava exploded onto the world market shelves in the early 1990s, the various

Oceanic cultural groups were quick to spot a commercial opportunity. Small-scale kava growers on Hawai’i increased production. Large-scale kava plantations appeared on New

Caledonia and even Guatemala, with less intensive attempts to cultivate kava elsewhere.

Everyone from consumers on the street to investors on Wall Street rode the kava wave.

Exports of kava rose dramatically from Pacific island nations. In a seven-year period from 1995 to 2002, Vanuatu saw kava exports rocket from US $432,000 to US

$4,527,000. But in 2002, reports claimed that a few people in Germany had suffered hepatotoxicity (liver damage) from kava supplements. Eight countries prohibited the sale of kava products. The United States did not, but issued a warning. The worldwide kava craze temporarily crashed (Lindstrom 2009). Germany and other countries later lifted their bans. Sales in kava products are recently again on the upswing (Ligo 2016).

Research has since attributed reported incidents of hepatotoxicity to the poor quality of the raw material that most likely contained mold, along with incorrect preparation (Boyle

2009; Teschke, Sarris, & Schweitzer 2012).

Kava is noteworthy here for two reasons. Among the many contemporary accusations of biopiracy, kava is one of the few, if not the only instance, of technology being able to trace a disputed medicinal plant to a relatively precise geographic location and time. The kava researcher Vincent Lebot genetically traced kava’s origin to islands in northern Vanuatu. There Lebot claimed Piper methysticum was first domesticated from

Piper wichmannii, a variety of local pepper, about 2,500 to 3,000 years ago (Lindstrom

2009). However, kava’s extensive cultivation has led to multiple varieties on the same island, plus islands across Oceania. There are nine varieties on Samoa alone. As Pacific

Islanders ventured across the Pacific Ocean, kava’s social importance from medicinal plant to recreational use was disseminated (Singh 1992).

Dutch explorers were introduced to kava in 1616 on Wallis and Futuna Islands. It took about 300 years for kava extracts to make their way onto the shelves of European pharmacopeias (Lindstrom 2009). Fifteen lactones (kavalactones) have been identified in the kava root, a tribute to its naturally powerful properties (Balick & Cox 1996). Yet until the 1990s, kava use was mostly relegated to Pacific Islanders. The first wave of kava

72 moving outward from its origin in Vanuatu did not generate accusations of biopiracy. The second wave in the 1990s did (Ji 2011; Lindstrom 2009; Singh 1992).

Unlike cinchona (at least until it was pirated by the British and Dutch), kava was willingly exported beyond its original area of cultivation by Melanesians and

Polynesians. Due to this, variations of the original kava have disseminated across the

Pacific Ocean. This is where the second instructive part of the kava controversy is relevant. It was reportedly the Hawaiian stock of kava that was used for commercial plantations during the kava craze. As sales of kava increased, so did accusations of biopiracy. The claims did not fall on a strict developing world-developed world axis as with most biopiracy disputes. Instead, concerns about kava biopiracy opened various levels of disputes among Pacific Island countries and among ni-Vanuatu (Ji 2011;

Lindstrom 2009; Singh 1992).

A good deal of the biopiracy disputes revolved around rights to cultivate and sell kava within Vanuatu. Attachments to kin and geographic locations are strong among the ni-Vanuatu. So is attachment to kava, which is embedded in the ni-Vanuatu culture. A kin or group may claim ownership to a specific variety of kava that may in turn be claimed by another kin or group of ni-Vanuatu. Such conflicting claims would be difficult to mediate. There are also cultural and gender restrictions regarding the use of kava. Therefore, designating a variety of kava as belonging to all ni-Vanuatu has the potential to create competing claims of ownership among regions or cultural classes, down to families or the individual (Lindstrom 2009).

Since it is believed kava’s use was modified by its introduction into Melanesia, as well as Polynesia (Singh 1992), sorting out what group (or even island) holds what

73 particular authority over indigenous knowledge of kava is impossible. The fears of outsiders poaching intellectual knowledge that drove promulgation of the CBD can also exist within developing countries and peoples.

A final brief example of another plant further demonstrates the difficulties in sorting out ownership of traditional knowledge. During the colonial period, Europeans noticed that enslaved Africans and Amerindian women used the peacock flower,

Pionciana pulcherrima, as an abortifacient to prevent children from being born into slavery. The plant is considered native to the West Indies. Yet what group, Amerindians or Africans, hold responsibility for realizing the plant’s properties is unclear. By the

1700s, West African medicinal knowledge had merged so effectively with Amerindian medicinal knowledge that little apparent difference remained (Schiebinger 2004).

Chapter Summary

As can be seen by these examples, sorting out who owns or originated traditional ethnobotanical knowledge is problematic much of the time, and impossible in most cases.

Supposed theft of intellectual property was a key driver for authoring the CBD. Biopiracy narratives create an Us-Them construction that leads the public to believe clear distinctions can be drawn with confidence. These examples have shown that this is not so. Accusations of traditional knowledge theft exist not only in a developed-developing world polarity, but also among indigenous groups.

In this respect, history has an odd way of determining villains and saints according to the temporal moment and political lens of their deed. Let us return to the example earlier in the chapter. Dioscorides is lauded for amassing and distributing ethnobotanical knowledge. Yet his interest in botany was piqued during his travels as a

74 young soldier (Osbaldeston 2000). Imagine if Dioscorides was compiling his work today, slogging through rainforests from village to village, inquiring about the medicinal uses of plants from indigenous peoples, and scribbling down every detail. Then, upon returning to his office, he published a volume of his findings. Dioscorides would probably be excoriated. He would be denounced as a biopirate.

CHAPTER SEVEN

THE CBD AND IMPACTS ON ETHNOBOTANICAL RESEARCH

The CBD in Practice and the Nagoya Protocol

Chapter Introduction

When the Convention on Biological Diversity (CBD) was promulgated in the early 1990s, there was optimism that indigenous peoples would become equal partners in the bioprospecting equation. The CBD was intended to reduce claims of biopiracy by providing a universally recognized framework of benefit sharing and research guidelines.

The coming years would show that good intentions are often derailed by real-world complications. Eventually, those problems would be largely addressed in the Nagoya

Protocol. But in those intervening 18 years, the CBD would at times create confusion instead of clarity, or impede rather than facilitate. In some cases, vague or conflicting guidelines would prove detrimental to both academic researchers and indigenous groups.

The CBD and the narrative of the pristine rainforest introduced a new set of actors into the biopiracy narrative—that of the Non-Governmental Organization. This chapter examines instances of some NGOs exerting onerous pressure on ethnobotanists and legitimate research projects. It explores whether some NGOs and environmental groups are operating to benefit their own objectives, rather than those of indigenous peoples.

The Convention on Biological Diversity: A New Era, New Problems

The Convention on Biological Diversity was heralded as a new day for the world of bioprospecting. The CBD was promulgated at the Earth Summit in Rio de Janeiro,

Brazil, on June 5, 1992. The modest document consisted of 42 main articles in 28 pages

(United Nations 1992). The CBD shifted traditional knowledge and plant resources from being a common heritage open to access by everyone, to being the property of countries

(Moran, King, & Carlson 2001). Additionally, it required that researchers obtain the prior informed consent (PIC) of indigenous communities before conducting research

(Schuklenk & Kleinsmidt 2006). The CBD was a culmination arising from a growing worldwide discourse on threats to global biodiversity, complaints by indigenous peoples of economic and cultural exploitation, and claims of biopiracy. The document was officially adopted on December 29, 1993, with 168 countries as signatories (United

Nations 2015b).

The CBD accomplished a great deal. Among its three primary objectives were: to conserve biological diversity; to ensure the sustainable use of the elements contributing to biological diversity, and; to provide for the equitable benefit sharing derived from the use of genetic resources (United Nations 2015a). Article 8 of the CBD encourages countries to establish and regulate protected areas for the conservation of biological diversity and to “ . . . preserve and maintain knowledge, innovations and practices of indigenous and local communities . . . ” in a sustainable manner (United Nations 1992, 6). It also calls for revenue sharing from such practices. Article 15 provides for state sovereignty over biological resources. Access to these resources must be “on mutually agreed terms” and

“subject to prior informed consent” with the entity providing the resources. Article 15 also calls for benefit sharing in a “fair and equitable way” (United Nations 1992, 9-10).

Although the CBD was working to help indigenous peoples, it left unresolved key components. This unwittingly created unforeseen impediments that caused confusion among both indigenous groups and researchers in certain instances. Differing opinions as to what entity is entitled to grant prior informed consent, and the difficulty in drawing lines on biological resources, allowed outside actors to shut down an ethnobotanical research project between an American university, a Mexican research institute, and an indigenous group in Mexico. It is important to note that these events took place before the

Nagoya Protocol that sought to address perceived insufficiencies of the CBD.

Ethnobotanical Research post-CBD: Hard Lessons in Chiapas

With the CBD in place, researchers moved to initiate benefit-sharing agreements.

There was confidence that research in indigenous communities could move forward unencumbered by accusations of biopiracy. That confidence soon disappeared. A new set of actors arrived on the scene with fresh allegations of biopiracy. They were international environmental NGOs claiming to act on behalf of exploited indigenous groups. One anti- bioprospecting NGO in particular used clever publicity to muster opposition against a bioprospecting research project in Mexico known as the Maya ICBG.

In 1998, Brent Berlin was the Graham Perdue professor of anthropology and

Director of the Center for Latin American and Caribbean Studies at the University of

Georgia, Athens (UGA). His wife, Elois Ann Berlin, was a medical anthropologist and an associate professor of anthropology at the same university. The Berlins were the recipients of a grant from the International Cooperative Biodiversity Groups program

(ICBG), administered by the Fogarty International Center under the auspices of the

National Institutes of Health (NIH) (Berlin & Berlin 2004; Rosenthal 2006).

The $2.5 million grant (over 5 years) was geared toward the following goals: locate, analyze, and evaluate local plants and other biological material for bioactive agents; identify and evaluate the medicinal and economic value of bioactive agents in plants for both local communities and global markets, including compiling a materia medica for the Highland Maya communities; produce an Ethnoflora of the Highlands of

Chiapas in the Tzeltal and Tzotzil languages; and bolster academic interaction between the UGA and the El Colegio de la Frontera Sur (ECOSUR), a research institute located at

San Cristobal de las Casas in the state of Chiapas, Mexico (Berlin & Berlin 2004).

The project was approved as a partnership between the UGA and ECOSUR. As part of the grant requirements Molecular Nature, Ltd. (MNL), a private biotechnology firm in

Wales, United Kingdom, was brought in to perform the laboratory analysis of plant extracts that were collected in the field. Upon the development of a commercially viable pharmaceutical product, the royalties were to be split between UGA, ECOSUR, MNL, and the Promotion of the Intellectual Property Rights of the Highland Maya of Chiapas

(PROMAYA). The latter was an NGO created by the Berlins to dispense royalties (if any) to local communities (Berlin & Berlin 2004; Hayden 2003). The project was known as the Maya ICBG. The grant project also invested US $80,000 in laboratory equipment for ECOSUR. Roughly 12 percent of the grant would be dedicated to training up to 24

Maya to assist with field research and laboratory duties (Belejack 2002; Brown 2003).

The Highland Maya in Chiapas State number over 900,000, reside in over 8,000 villages, and include four languages (see Figure 9). Brent Berlin has performed research

79 in the region for over four decades. He is fluent in both Spanish and Tzeltal Maya (Berlin

& Berlin 2003; Brown 2003; Cragg, Katz, Newman, & Rosenthal 2012).

Following PIC guidelines as outlined in the CBD, the Berlins obtained consent of the local community leaders and members through a three-month long process of informational meetings held at villages. They also distributed flyers and aired radio announcements in native languages. The community meetings included theatrical plays in local languages that explained the project’s purpose. Goals, methods, and the unlikelihood of any revenues being generated were emphasized. Additionally, progress

Figure 9. Chiapas State in Mexico was the region where a legitimate research project was derailed by outside NGOs. Chiapas (colored in pink on basemap) inset map from Berlin & Berlin, 1996, and A.C. Procomith, 1991; Basemap from ESRI, 2014; Mexican state boundaries shapefile from Hoel, 2013; country boundaries shapefile from Natural Earth, 2016.

80 was continually updated on the UGA website. Of the 15 municipalities home to 47 villages (parajes) asked to participate, 46 agreed (Rosenthal 2006). This included about

30,000 people (Berlin & Berlin 2003). By 1999, the Berlin’s Maya ICBG project was up and running. Almost immediately there were claims of biopiracy.

A group of local healers operating under an NGO later known as COMPITCH

(Consejo Estatal de Medicos y Parteras Indigenous Tradicionales de Chiapas) voiced opposition to the agreement, claiming the agreements secured by the Maya ICBG were not valid and that the Berlins had been deceitful about the economic potential of the project (Berlin & Berlin 2004). Critics of the Maya ICBG contended that the project had not properly informed local communities on the international discussion about traditional knowledge and the patent process; claimed that the Maya ICBG had not collected the required number of signatures from community members; and insisted that the agreement include input from all two million Maya-speaking peoples in Mexico and Guatemala

(Rosenthal 2006).

In normal times, these complaints may not have been enough to halt the project.

But Chiapas State in the late 1990s was in the middle of political upheaval. On January 1,

1994, an uprising focused world attention on the plight of the Maya and frustrations over the recently approved North American Free Trade Agreement (NAFTA) (Froehling

1997). Anger toward the Mexican government over a history of social inequities made the Maya ICBG an easy target. Lack of a recognized regional authority in Chiapas made it easy for outside NGOs to fill the power vacuum (Cragg, Katz, Newman, & Rosenthal

2012).

The most vocal NGO was the Rural Advancement Foundation International, now known as the Action Group on Erosion, Technology, and Concentration (RAFI/ETC), based in Canada. Patrick Mooney, ETC’s director, is generally recognized as having first used the term “biopiracy” in the early 1990s (Berlin & Berlin 2004; Hamilton 2006b;

Hayden 2003; Robinson 2010). Global Exchange, a San Francisco-based NGO, also attacked the Maya ICBG project on the legitimacy of the agreements signed by the participating Maya communities (Berlin & Berlin 2004).

RAFI (ETC) used various media platforms to engineer a successful publicity campaign that discredited the validity and purpose of the Maya ICBG. Claims of biopiracy, the patenting of biological resources, and theft of traditional knowledge captured the attention of the global media. RAFI cleverly likened the Maya ICBG to past fears of Yankee colonialism in Central America (Berlin & Berlin 2003; Berlin & Berlin

2004; Brown 2003; Rosenthal 2006). At one point, Brent Berlin was likened to pukuj, a local incarnation of Satan (Belejack 2002; Brown 2003).

In a news release, RAFI (1999) announced that some indigenous organizations in

Chiapas “ . . . claim that their indigenous knowledge and resources are being stolen” (2).

There were additional complaints by project opponents that, by setting up PROMAYA, which would disburse monies to local communities, it was a sign that the Berlins were acting without properly consulting the Highland Maya (Rural Advancement Foundation

International 1999).

Claims that researchers had side-stepped proper channels to obtain prior informed consent drove a good deal of the controversy. The Convention on Biological Diversity does not define PIC (keeping in mind this was pre-Nagoya). It only stipulates PIC be

82 obtained and defers to individual nations on the process (United Nations 1992, 9-10).

Compounding this was a lack of a universal definition for prior informed consent

(Firestone 2003). This became a major problem for the Maya ICBG.

Countries like the Philippines and Peru have established detailed requirements on obtaining PIC, while requirements in Mexico (at least at the time) are less explicit (Berlin

& Berlin 2003). Therefore, in an effort to adhere to CBD guidelines, the Berlins followed

Mexican law when they obtained consent. Article 87 BIS of Mexico’s General Law of

Ecological Balance and Environmental Protection states that authorization to exploit

“wild flora and fauna . . . shall only be granted with the . . . informed consent of the owner or legitimate holder of the land where the biological resource is located” (Berlin &

Berlin 2003; Federal Official Gazette 1998, 53; Hughes 2002). Yet from what entity that consent should be sought is problematic for researchers, particularly in Mexico. Nor was the CBD any help. The CBD uses the generalized term “contracting party” when discussing bioprospecting agreements (United Nations 1992). This highlighted another weakness of the CBD. The CBD does not specifically define state or community (Aguilar

2001; United Nations 1992). Without a universal definition of community, researchers may have trouble identifying at what level within an indigenous community they should seek consent. Much of the debate centered on interpretations of community in Maya culture.

Establishing the proper geographic boundaries, physical, cultural, or political, that define an indigenous community is extremely difficult due to the unique socio-political structure that each indigenous group possesses from country to country (Berlin & Berlin

2003). Communities may not always be defined by language or culture, but by region.

Additionally, adversarial relationships of an indigenous people to the controlling national political leadership, or even amongst groups, can be problematic in defining a community

(Rosenthal 2006). The region encompassed by the Maya ICBG was particularly challenging.

The parajes (villages) are considered by many to form a community in the Maya

Highlands. A history of independence at the paraje level, coupled with a weak municipal system in this part of Mexico, places decision-making in the hands of local individual communities (Rosenthal 2006). Yet there is no nationally recognized system of communities in Mexico, according to Neil Harvey at the University of Mexico. He notes that obtaining PIC from indigenous communities in Mexico is difficult due to the absence of a universally accepted definition of community (Berlin & Berlin 2004). Schuklenk &

Kleinsmidt (2006) acknowledge that the Maya communities do not have nor speak with a single, unified voice.

Although the Maya ICBG obtained consent at the community level (Berlin &

Berlin 2003), COMPITCH and RAFI argued that by only securing the permission of some Maya communities, and not all of them throughout Mexico and Guatemala, the

Maya ICBG was co-opting the traditional knowledge of those groups not participating

(Brown 2003). That is, since a particular plant in Chiapas might grow in another country, and be used by groups there, permission was needed by all groups who might possibly encounter the plant.

This was another weakness of the CBD as it applied to the Maya ICBG: how to delineate boundaries of a natural or biological resource. This revisits Article 87 BIS of

Mexico’s General Law of Ecological Balance and Environmental Protection that allows

84 exploitation of biological resources with the permission of the landholder upon whose land such resources are encountered (Federal Official Gazette 1998). As previous chapters have shown, it is possible to find the same medicinal plant in numerous countries. Thus, the difficulty of drawing borders on biological resources can be as difficult as delineating boundaries of traditional medicinal knowledge.

Extending the demands made on the Maya ICBG researchers, it is easy to see how research could be hamstrung if a researcher wanted to study a plant found in a particular region of one continent, when the plant grows on multiple continents. To have accepted

RAFI’s demands would have entailed collecting permission from all Maya communities throughout Central America, an impossible task that would have effectively ended any bioprospecting—which was what RAFI and COMPITCH wanted. At one point, RAFI

(ETC) stated, “ . . . RAFI regards all bioprospecting agreements to be biopiracy” (Brown

2003, 122).

The anti-globalism protest that helped fan opposition to the Maya ICBG became a child of globalization during the course of the protest. Although opponents criticized the

Maya ICBG for using individual Maya communities as legitimate representatives of all

Maya people, groups like COMPITCH had no problem with speaking for all Maya communities. However, none of the Maya communities participating in the project were part of COMPITCH’s opposition (Rosenthal 2006).

The complaints by RAFI and COMPITCH were found to be contradictory.

Canada’s Indigenous Caucus reviewed the dispute and noted that RAFI and COMPITCH simultaneously insisted for community autonomy, while insisting that the Mexican government enact laws addressing bioprospecting in the case (Brown 2003). By 2000, the

85 dispute had reached a point where the Mexican government tried to mediate a solution.

The arbitration ended with government representatives claiming that COMPITCH was refusing to negotiate ethically (Berlin & Berlin 2003). The Maya ICBG ended in October

2001, when ECOSUR removed itself from the project due to the controversy. Without a sponsoring Mexican institution, the NIH withdrew funding (Brown 2003; Rosenthal

2006).

The communities in the Maya ICBG project were co-opted by outside NGOs due to the inherent makeup of these communities that did not possess a recognized authority that could speak for all of the communities (Rosenthal 2006; Schuklenk & Kleinsmidt

2006). COMPITCH, a native organization, was itself seen as being the mouthpiece of outsiders who manipulated the situation to their own political agenda (Brown 2003).

The Convention on Biological Diversity was intended to empower indigenous peoples by giving them control over biological resources. In the Maya ICBG case, the

CBD acted to de-power these groups. The Maya communities that could have benefitted culturally, educationally, and economically, had control wrested from them by outside actors who returned to wealthy developed countries.

The CBD in the New Century: A Work in Progress

Impact of the CBD and the Biopiracy Narrative

While the CBD accomplished much, it also left certain concerns unresolved. This included insufficient guidelines on what formed equitable benefit sharing, and a lack of clear and universal guidelines on how access to biological resources should be granted and regulated. Overly stringent laws, seemingly interminable negotiations, and confusion over what entity could grant access, at times created frustration on the part of researchers

86 following adoption of the CBD. Countries and indigenous groups possessing biological material became disillusioned when benefit sharing deals did not generate huge sums of money (Cragg, Katz, Newman, & Rosenthal 2012). As seen with the Maya ICBG case, it was not just varying interpretations of the CBD’s components among researchers and governments, but also outside groups that helped create confusion and frustration.

By some accounts, this caused research to flounder (Lewis 2003). Researchers saw little purpose in approaching countries with bioprospecting projects, as there was reluctance to permit projects in a “politically charged atmosphere” (131).

The Nagoya Protocol: Bandage or Cure?

By September 2002, a little over 10 years after the CBD was adopted, there was a desire on the part of both developed and developing countries to improve upon the CBD.

Eight years later, in October of 2010, the Nagoya Protocol was adopted. It became effective on October 12, 2014 (Schindel et al. 2015).

The Nagoya Protocol was seen as necessary to diffuse the growing frustrations felt by all parties. Frameworks for granting PIC were judged to be inconsistent, if applied at all. Some countries did not possess the ability to develop the necessary laws. There was suspicion on both sides. Some developing countries believed that agreements would be ignored once the biological material left their country. Researchers were confounded by byzantine regulations, delays in negotiating, and absence of a legal framework, which would leave them open to charges of biopiracy (Burton & Evans-Illidge 2014).

The Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable

Sharing of Benefits Arising from Their Utilization to the Convention on Biological

Diversity provides for legally-binding guidelines designed to eliminate some of the issues that had proved disruptive to the Maya ICBG.

The Nagoya Protocol establishes “international access standards” (Buck &

Hamilton 2011, 47) that are aimed to provide more consistency and clarity in obtaining

PIC and MAT (mutually agreed terms). This will ensure that indigenous or local groups have the ability to grant access to biological resources to which they have a previously established right to control. The Nagoya Protocol also clarifies obligations for benefit sharing (Buck & Hamilton 2011).

Full discussion of the Nagoya Protocol is beyond the scope of this chapter. It is too early to judge how the new guidelines will pan out in practice. Some have expressed concern that the Nagoya Protocol, like the CBD, fails to clarify certain key terms and concepts, such as “indigenous and local communities,” or “traditional knowledge associated with genetic resources,” or what it means for knowledge to be “held” by a community (Buck & Hamilton 2011, 54-55).

Bioprospecting in the Digital Age

There is another bioprospecting method that has been gaining popularity but remains outside the regulation of the CBD or other legal guidance. Searching ancient texts for bioprospecting clues is now being used by some as a way to circumvent onerous bureaucratic regulations. It is also an indication of how modern technology has expanded methods used in bioprospecting.

Ancient texts are scanned and then searched by a computer for references to medicinal plant usage. These computer programs identify concepts and expressions, such as “leaf of a tree.” Such phrases provide important clues about medicinal uses, although

88 identifying the exact plant can be problematic. Computer software can now scan up to

1,200 pages an hour, greatly facilitating what was previously a time-consuming process

(Buenz, Schnepple, Bauer, Elkin, Riddle et al. 2004).

This method has already yielded a success. The Ambonese Herbal, authored by

Georg Rumphius in 1692 (the first of several drafts), was scanned by a computer. A treatment for dysentery was found by using the nut of the atun tree (Buenz, Bauer,

Johnson, Tavana, Beekman et al. 2006).

For those wishing to protect the knowledge of indigenous peoples, this raises a troubling question. If indeed traditional knowledge can be mined from ancient texts and a successful pharmaceutical actually be developed, who is deserving of benefit sharing?

Potentially, this could involve the TEK of a group that no longer exists. Or if present-day descendants exist, are they entitled to benefit sharing, even though they no longer reside on the land or practice TEK? Scanning old texts for possible drug leads may seem inconsequential now, but if a commercially successful pharmaceutical is developed, such dilemmas will be relevant.

Chapter Summary

The Convention on Biological Diversity, which was heralded by many as the beginning of a new era, in fact became just that. Ambiguous guidelines confounded researchers. In the case of the Maya ICBG it allowed outside actors to exploit indigenous people for their own agenda. This led to the perceived shortcomings of the CBD being addressed in subsequent years.

By then new biotechnology had appeared. Microbial organisms were gaining attention. Further, the prime areas for these microbes appeared to be out of the reach of

89 confounding documents like the CBD. In a span of a few years, bioprospecting expanded to include microbial organisms. The CBD, despite its laudable goals, had partly contributed to this new era.

CHAPTER EIGHT

THE PHARMACEUTICAL INDUSTRY

From Rainforest to Pharmaceutical: The Long, Uncertain, and Expensive Road

Chapter Introduction

The rationale that traditional medicinal knowledge is being used as a roadmap by stealthy pharmaceutical companies to first identify medicinal uses of plants, then use that knowledge to create a wonder drug that denies ownership and profit to indigenous peoples or developing countries, is central to contemporary claims of biopiracy. This chapter follows the process of drug development from field to lab to pharmaceutical counter. It demonstrates that drug development from plants is neither quick, nor easy, nor cheap.

Claims of biopiracy arose due to a belief that pharmaceutical companies create structures to cheat rather than to profit alongside indigenous peoples in drug development. This chapter examines what happened to a company that was built upon the principle of benefit sharing with indigenous peoples.

Another accepted narrative is that traditional ethnobotanical knowledge provides pharmaceutical companies with a clear roadmap to generate wonder cures. This chapter reveals that relying on TEK can be helpful in some instances, but in the complex process of pharmaceutical development, success is dependent upon many factors.

Green Gold Reborn

The technique of turning a plant into a pharmaceutical is not a recent development. An English physician named William Withering was experimenting with digitalis by 1775. Withering learned that folk healers used Digitalis purpurea as a remedy for dropsy. Dropsy is when the body swells due to inadequate circulation by the heart. Withering administered a mixture of ground D. purpurea leaves and water to patients. Even with his crude mixture Withering had a startling success rate of 65 to 80 percent (Cox 1994). And that was before the advent of modern pharmaceutical technology that can isolate compounds not imagined by Withering.

Still, two hundred years after Withering’s rudimentary experimentations, the road from plant to useful pharmaceutical remains a difficult, questionable, and expensive process. Hopes in modern technology’s ability to develop useful pharmaceuticals from plants has been dashed, revived, and dashed again.

Starting in the 1950s, pharmaceutical companies placed greater emphasis on synthetic chemistry (Hayden 2003). The urgent necessity for antibiotics during World

War II, coupled with the need to combat numerous afflictions during the post-war years, had tantalized the pharmaceutical industry with the sizeable profits to be reaped from large-scale drug development. Random mass screening of plants and biological material was initiated (Petrova 2014).

The mass screenings created a new problem. The thousands of compounds isolated created a bewildering backlog when it came to the manual testing of these bioactive agents in test tubes. This was because the biochemical processes and molecular dynamics underlying the effectiveness of drugs was largely beyond the grasp of

92 researchers at the time. As a result, massive libraries of compounds were compiled with little hope of ever being thoroughly tested for drugs. Identifying a compound with therapeutic potential was often dictated by chance (Petrova 2014).

The peak excitement for plants-to-medicines lasted only about seven years, from

1953 through 1960. Reality was a hard teacher. During a 31-year period starting in 1950, just nine drugs (including contraceptives, antihypertension, tranquilizers, and vincristine and vinblastine) were developed from bioactive extracts in plants. An additional 40 steroids were also developed from plant extracts during this period (Farnsworth &

Soejarto 1985).

Starting in the 1970s, pharmaceutical companies lost interest in screening plant samples from tropical locations. Biotechnology began to make advancements that spurred hope that the massive libraries of compounds acquired through mass screenings would create a new era of synthetic drug production (Sittenfield & Gamez 1993).

Spending by pharmaceutical companies for research and development of plants- to-drugs declined rapidly. By 1974, just one pharmaceutical company in the United States was engaged in screening plants as a source of medicines. The amount invested in that research was estimated at a mere $200,000 (Farnsworth & Soejarto 1985).

Economic considerations also had an impact on the scaling down of plant-to-drug research. A period of corporate mergers led to the global drug market being dominated by a handful of multinational pharmaceutical firms. The vertical integration of pharmaceutical companies created enormous pressure for research to produce profitable drugs (Laird & ten Kate 2002, ). These economic considerations cooled enthusiasm for expensive plant-to-drug research programs.

By the early 1980s, new techniques and advances in biotechnology, and specifically genetic engineering, ratcheted up excitement over potential successes in synthesizing drugs (Petrova 2014). By the 1990s, innovative biotechnology techniques were opening a new era in drug research.

Techniques known as combinatory chemistry and mass bioassay screenings were part of the shift away from the pharmaceutical industry’s emphasis on medicinal plants.

Combinatory chemistry involves mass screenings of thousands of (synthetic) compounds at a time. If a lead (a useful compound) is identified, pieces of it are removed or added to in a process called combinatory chemistry until a compound that can be synthesized is produced. Yet performing this procedure on compounds extracted from plants is problematic. Many natural compounds are too complex to be reproduced in the lab

(Conniff 2012).

Plants never receded far from the attention of researchers. Renewed interest in them soon returned. Researchers were concerned that they may be running out of bioactive compounds that could be reproduced synthetically in the lab. As technology increased, identifying useful bioactive compounds in plants became more accurate and less expensive (Newman 1994). Once again, it seemed plants could be useful to pharmaceutical development.

Earlier in 1985, one development played a major role in reviving interest in plants as a source of medicines. That was the estimated economic value of plants as medicines in what could be loosely called Farnsworth’s list (see Appendix A).

Farnsworth and Soejarto (1985) found that during the period 1959-1973, an average of 25.3 percent of pharmaceutical prescriptions in the United States had their

94 core bioactive compounds extracted from plants. By extrapolating that number against the cost of drug prescriptions, the pair estimated the 1980 value of pharmaceutical prescriptions derived from plants to be $8.1 billion. When over-the-counter sales of plant- based medicines was included, this pushed the figure up to $9.8 billion. Five years later in 1985, that number stood at around $18 billion (Principe 1991).

Farnsworth followed up the groundbreaking article with another one shortly after.

The latter one pointed out that out of 119 pharmaceuticals derived from plants, 88 were based on leads generated through traditional medicinal knowledge (Farnsworth, Bingel,

Soejarto, & Guo 1985). Farnsworth and Soejarto (1991) generated more interest in medicines from tropical plants, by noting that 95 plant species accounted for 121 plant- based drugs. Additionally, 38 plant species found in tropical rainforests were responsible for 45 drugs.

With pharmaceutical prescription costs on a steady linear rise, and only about

5,000 plants at the time having been analyzed for bioactivity out of a possible 500,000 species (Farnsworth & Soejarto 1985), researchers took notice. It probably helped that

Farnsworth was part of the initial research team at Eli Lilly that developed vinblastine and vincristine (of the rosy periwinkle fame) into successful anti-cancer drugs (Brown

2003).

These articles helped fuel the growing attention on the rainforests as repositories of potential miracle drugs. The thinking went that, if 74 percent of just over 119 drugs were based on traditional knowledge leads (Farnsworth, Bingel, Soejarto, & Guo 1985), then certainly indigenous peoples residing in biologically diverse areas such as the

95 tropical rainforests must be sitting on a green gold mine. Examining the process of bringing a pharmaceutical to market offers a sobering perspective.

The Myth of (Easy) Green Gold

The true cost of drug development from plants is hard to pin down. Common figures range from as little as $50 million to $500 million with the process requiring up to

18 years (Aylward 1995; Brown 2003; Farnsworth & Soejarto 1991; Laird & ten Kate

2002; Moran, King, & Carlson 2001; Reid et al. 1993; Sittenfield & Gámez 1993). Others have estimated that the process of developing a potential drug, starting with collection of plants through research and clinical trials, ending with placement on market shelves as pharmaceutical, generally takes 10 to 15 years at a cost of $300 million to $2 billion

(Moran, King, & Carlson 2001; Rose, Quave, & Islam 2012). A more recent estimate put the cost at $1.3 billion (Petrova 2014). Whether the cost of development be at the low end or the high end of those figures, turning a plant extract into a successful drug is an expensive endeavor fraught with a myriad of possibilities for failure.

The generally accepted success rate starting with testing a plant for a bioactive compound, refining, and synthesizing a compound (if found), then developing and testing the drug during clinical trials, to getting final FDA approval, is a daunting one in 1,000 to one in 10,000 (Fabricant & Farnsworth 2001; Firn, 2003; Hayden 2003; Kumar & Tarui

2007; Newman 1994; Sittenfield & Gámez 1993; Petrova 2014).

Weiss and Eisner (1998) offered a more sobering analysis. Should a plant survive the generally accepted one in 1,000 chance of passing an initial bioactive screening to become a hit, only three percent of those hits survive further screening to become a lead.

Only about 10 percent of leads become compounds of interest and are selected for further

96 refinement and testing as potential drugs. Of that group, only 15 percent reach the market. In the end, of all the raw biological samples that make it through initial screening and go on to survive further testing as leads, development, and approval, only one in four million become a marketable drug.

Upon investigating the potential use of a plant for a drug, researchers face numerous questions. What will be the outcome of the drug in usefulness and safety? Can the necessary compounds be reproduced in the lab? Is the usefulness of the drug going to guarantee future profits to recoup research and development investments? Is the drug patentable? (Firn 2003).

The process starts with plant collection in the field. Different collecting methods are used to discover leads in plants. Ethno-directed research is one such approach (Balick

& Cox 1996). It is probably the most well-known. A second method referred to as random is not actually random. This relies on collecting plants based on non- ethnomedicinal traditional knowledge, yet suspected of having potential for bioactive compounds. This includes collecting a plant from every plant family within an ecological region to ensure the most genetic diversity (Ghorbani, Naghibi, & Mosaddegh 2006;

Miller 2011).

Once the field samples are shipped to the lab, the screening process is broken into two stages: first securing a hit, then a lead. A hit is a compound with an established structure that registers bioactivity against its target (Soejarto 1996). Screening for hits can be dependent on the part of the plant collected. Flowers can have a dramatically different chemical composition than roots, roots can be different than leaves, and established leaves different than new plant growth (Balick & Cox 1996). Extracts that show a hit or a

97 positive sign of a bioactive agent are dereplicated. Dereplication identifies the structure of new compounds from previously identified compounds. This is an important point.

The screening process is not geared solely for the elucidation of bioactive compounds, but novel compounds that have not been previously identified (Miller 2011).

Hits that show further potential are subjected to more analysis that isolates their chemical structures. A lead occurs when an identified compound possesses a distinct purity with a potent and discrete activity, and has a well-defined chemical structure connected to a specific activity that makes it suitable for a bioassay (Cordell 2000).

Two problems arise with the identification of a lead. Although plants may have a lot of bioactive agents, the effectiveness of those agents may be so weak as to make them useless as a pharmaceutical. When a lead is generated, further analysis may reveal that the chemical structure is so complex that laboratory replication into a synthetic drug is impossible (Firn 2003). Harm to consumers is another reason that many drugs get shelved in development. About 30 percent of compounds in development are too toxic for further development. An additional 30 percent of compounds are shelved due to lack of effectiveness as a drug. Other problems that derail promising drug compounds are related to the rate and length of effect, or problems ranging from absorption by the body (Petrova

2014).

A favorite past process was to use ethanol and methanol to extract compounds from raw plants. This method could disguise the potential of new compounds unless they happen to be extraordinarily powerful or abundant. This could be a contributing reason for the surprisingly low rate of new compounds identified during the 1980s and the early

1990s. To counter this, new technologies using nuclear magnetic resonance were

98 invented to fractionalize the compounds so that each compound would be better represented. Other modern technologies alternatively employ liquid chromatography or spectroscopes. By the 1990s, automated bioassays were capable of screening 30,000 to

300,000 samples per year. (Miller 2011). But more expensive technology does not guarantee success.

Although the aggregate number of secondary or specialized metabolites produced by all species of plants is estimated to be at least 200,000 (Pichersky & Lewinsohn 2011), in-vitro bioassays and mass screenings in the 1990s did not meet expectations of an increase in identifying novel bioactive compounds that led to new drugs. Researchers speculate that this was partly due to the redundancy of secondary compounds in plants, or that numerous secondary compounds come from a select group within a species, or are limited to just one species. Further, the drying of plants to prepare them for study changes the chemical makeup, potentially causing some compounds to become lost (Miller 2011).

Identifying a new compound leads to new problems. Initial testing requires 0.5 to

2.0 kilograms of plant material. If bioactive agents are identified and further testing is required, then upwards of 100 kilograms of plant material is required for drug production and synthesis (although modern technology is rapidly reducing this amount) (Balick &

Cox 1996; Soejarto 1996). This raises the question of obtaining enough raw source material.

Take the case of the infamous rosy periwinkle. The alkaloids vincristine and vinblastine both occur in such minute concentrations that a minimum of 250 kilograms of rosy periwinkle leaves is required to produce 500 milligrams of the cancer-fighting drug

(Balick & Cox 1996). The FDA recommended first dosage of vinblastine for adults

99 suffering from Hodgkin’s disease is 3.7 mg/m2 bsa (body surface area). That dosage may be increased up to a maximum of 18.5 mg/ m2 bsa (Drugs.com 2016). It is important to note that the dosage can vary slightly according to the the patient’s condition or medical supervision. Using the Standford University body surface area calculator, a male adult

5’10 tall and weighing 170 pounds has a bsa of 1.95 m2 (Standford Department of

Medicine 2016). That means a typical first dose for a pateint of this sex, height, and weight would be approximately 7.22 mg of vinblastine. Given that the dosage may be increased over time, and that thousands or millions of patients may require similar doses, it is easy to see how obtaining the necessary plant volume for producing a drug could become a barrier to drug development.

The United States has a rigorous and exhaustive drug approval process. The development of pharmaceuticals follows four basic stages. Once a drug is produced it is tried on animals to determine effectiveness and toxicity levels. A pharmaceutical company then files for an Investigative New Drug (IND) application. The preclinical trial then moves to Phase 1 where the drug is tested on human volunteers. In Phase 2 the drug is tested on actual patients to further fine tune the chemical composition and dosage. In

Phase 3 the safety and effectiveness of the drug is validated prior to approval for marketing. This process requires 4 to 7 years. At the end of Phase 3 the pharmaceutical company can request the Food and Drug Administration approve the new drug. If the application is approved, Phase 4 is required to scrutinize the drug’s safety and side- effects during an extended period of testing (Cordell 2000). There are other factors that make drug development a risky business.

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The patent clock on a new chemical entity (NCE) starts ticking soon after a compound is identified early in the screening process. A patent expires in 20 years, so a pharmaceutical company has a limited window to recoup its investment. If exclusivity rights are awarded, depending on the drug a pharmaceutical company may have up to 23 years of protection (Farnsworth & Soejarto 1991; Petrova 2014; Reid et al. 1993).

Whatever the cost of drug development, pharmaceutical companies operate on a deadline.

Then there is the small percent of drugs that recoup the investment of research and development. Only three out of 10 drugs that reach the market return a profit

(Aylward 1995). Clearly, pharmaceutical development from plants is not the quick and easy road to riches many might assume.

The Value of Traditional Ethnobotanical Knowledge in Drug Discovery: Map or Maze?

The biopiracy narrative has portrayed indigenous knowledge as extremely useful in identifying potential new compounds. This section shows the actual value of traditional knowledge in drug discovery is much less certain.

Interest in traditional healing practiced by indigenous peoples was gaining acceptance years before the CBD took effect in 1993. The World Health Organization

(WHO) acknowledged the importance of traditional medicine with the Primary Health

Care proclamation of Alma Ata in 1978. The Maastricht Declaration was promulgated by the World Congress on Medicinal and Aromatic Plants for Human Welfare in 1992. It recognized bioactive agents in plants and biological materials formed an important component of traditional medicine (Kim 2005).

On the heels of Farnsworth’s list in the mid-1980s, there was anticipation that

TEK contained the future pharmacopeia of mankind. The CBD was adopted to prevent

101 the theft of traditional medicinal knowledge and allow indigenous peoples to share the wealth of drugs-from-plants. We have just seen how the odds of a plant becoming a successful pharmaceutical are akin to winning the lottery. So which is it? Are pharmaceutical companies plundering the TEK of indigenous peoples? Or has the value of traditional medicinal knowledge been vastly overrated? Like the drug discovery process, this has no clear answer.

The use of a medicinal plant by indigenous people often leads to a dead end when it is attempted to be modified into a drug for populations in developed countries. Or it may lead to a completely different use, such as tubocurarine (as a muscle relaxant).

Sometimes the natural compounds in plants will lead to synthetic compounds that possess medicinal value. It is rare that drug research based on indigenous uses leads to a drug with the same uses in developed countries, such as quinine (Schultes 1994).

Indigenous communities may vary in their uses for the same plant (Schultes

1991). Communities may vary in ethnobotanical knowledge due to different diseases, diets, cultural, or physiological factors (Firn 2003). Thus, basing research on the traditional medicinal practices of one group can misdirect researchers.

Nor do all indigenous groups maintain a similar sized pharmacopoeia. The number of plants used in traditional healing varies widely even among groups in the same geographic area (Schultes 1991). Generalized predictions on the usefulness of a region’s biodiversity, or the traditional knowledge of any one group as an indicator of potential drug discovery, is problematic.

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Even though 80 percent of the global population relies on plants for medicine,

Plotkin (1991) points out that this may not be a vote so much for the effectiveness of traditional medicine, but rather a lack of access to better and more expensive medicine.

There is also the economic hurdle of obtaining the expensive and sophisticated screening equipment found in research facilities of developed countries. This is why plant simples are common in indigenous medicines. Plant simples are one or two plants, mixed or boiled without additional ingredients (Schultes 1991). Plant simples are convenient for indigenous healers but not for commercial pharmaceuticals. Synthesizing plant compounds is necessary as it is not practical to ship an entire plant to the market shelf in worldwide mass distribution.

Diseases common in the developed world are not generally commonplace in developing communities. Thus, plants may be used to combat different illnesses (Firn

2003). Plant simples used by traditional healers are utilized mostly for medical issues involving the skin, stomach and intestinal tract, inflammation, obstetric and gynecological problems, snakebites, or bleeding. Traditional medicinal practices are not used to treat cancers or complex, internal Western diseases. Conversely, Western medicine concentrates on medical issues that predominate in Western societies such as cardiovascular disease, nervous system disorders, abnormal tissue growths (neoplasms and cancer), or microbial illnesses (Balick & Cox 1996; Firn 2003). This is not surprising, as indigenous peoples can more easily diagnose external ailments like skin diseases, but lack technology to diagnose internal disorders (Balick & Cox 1996).

These differences in the medicinal uses of plants creates a dilemma for arguments of modern biopiracy. If indeed pharmaceutical companies are hijacking the knowledge of

103 indigenous medicinal knowledge, should not the Western uses of a plant’s healing powers mimic those found in indigenous communities? This is clearly not the case.

However, that belief was partly effective in pushing bioprospecting toward a new era of microbial bioprospecting.

Economics play a major factor in the value of traditional knowledge. It is a sad fact that diarrhea kills five million infants each year. Yet because diarrhea and other diseases like malaria or schistosomiasis are largely developing world afflictions, little drug research is devoted to finding potential cures (Cordell 2000). Pharmaceutical economies of scale are therefore a driver in drug research (Laird & ten Kate 2002).

Economic pressures drive drug research in countries like the United States, while indigenous healers are under no such pressures (Balick & Cox 1996). Of 50 drugs whose initial field research began as tips from indigenous medicine, 22 percent resulted in drugs used to fight cardiovascular disease, while only 2 percent were used by indigenous peoples for similar diseases (Balick & Cox 1996). Thus, funding often determines the direction of research, which is almost always targeted at afflictions of developed countries. Figure 10 shows a sampling of some of the most well-known drugs derived from traditional ethnobotanical knowledge. Appendix B provides a table on 50 drugs derived at least in part from TEK.

Studies are mixed as to the odds of success between basing initial pharmaceutical research on indigenous knowledge or stochastic screenings. Balick and Cox (1996) noted that a general historical overview of plant screening based on ethno-directed observations is more likely to generate some level of success as opposed to more random methods.

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Figure 10. Drugs discovered from ethnobotanical leads. Some of the most well-known drugs developed were based on at least partial traditional ethnobotanical knowledge. See Appendix B for a complete list. Image from Balick & Cox, 1996.

Some studies found focusing on plants with an established medicinal use among indigenous peoples returns a higher success rate in the screening process. Holmstedt and

Bruhn (1995) noted that as far back as 1976, a study by the National Cancer Institute showed a higher success rate for anti-cancer tumor screening based on ethnobotanical observations versus random screenings. That was before advanced biotechnology became common place in the laboratory.

Basing drug development on ethnobotanical leads appear to be useful against certain ailments, such as combating antifungal and antibacterial afflictions, along with hypoglycemic ailments (Balick & Cox 1996). A study led by Cox (1994) found a distinct difference between ailments and diseases treated with medicinal plants by indigenous healers, and drugs used in developed countries that were derived from ethnobotanical

105 leads. Some speculate that the economics in drug use between wealthy and poor societies may leave ethnobotanical leads under utilized by researchers (Firn 2003).

Caution is prudent in reading too much into studies that show a high initial bioactivity for the screening of traditional medicinal plants. Miller (2011) points out that some previous studies that have compared the success rate generated by ethnobotanical leads versus mass screening are based on initial screenings. The chance of a hit or lead becoming an effective drug is another matter. Miller makes another important observation that there are different interpretations of “random” collection. Plants may be collected without ethnobotanical guidance, but may still be collected based on other information such as taxonomy.

Different cultures may return different success rates (Balick & Cox 1996).

Farnsworth (1994) argued that for traditional medicinal knowledge to be relevant in pharmaceutical research, data on indigenous medicinal practices must be meticulous, otherwise the process reverts back to a shotgun approach. Utilizing different collection methods should be adjusted according to the goal of the program (Miller 2011).

It remains unclear how important TEK/TMK is to the pharmaceutical industry in drug development. While some repeat the mantra that plants serve as the basis for one in four drugs, others point out the odds that only one in 10,000 compounds becomes a marketable pharmaceutical (Hayden 2003). This uncertainty explains the historical rollercoaster of interest in drugs-from-plants. Excitement reaches a peak, only to have hopes plummet, then a new technology causes interest to be reborn.

By 2002, an additional 16 plant-based drugs had been added to Farnsworth’s original list. This raises the number of drugs based on plants to 135. Although upwards of

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60,000 plant species have been screened for drug potential, many of those screens were for specific diseases. Since there is agreement that about 2,000 species have been exhaustively screened for bioactivity, this leaves a tantalizing possibility of 350,000 species left to screen. Based on the formula:

135 # &' 1/34 )560 7&)0.3)0- 5+ 8*,3/0 = # &' ()*+,- -./00+01 # &' -001 ()*+,- − # *)/0*1: -./00+01

Miller (2011) conducted estimates on the number of potential drugs to be derived from plants.

Plugging 2,000 into the equation for the number of plants thoroughly screened returns a possible 20,115 to 23,490 potential undiscovered drugs, based on 298,000 to

348,000 species left to examine. Yet if the upper number of 60,000 plants screened for bioactivity is used, and the number of plant species remaining to be screened is 240,000 to 290,000, then 540 to 653 new drugs are waiting to be identified by bioprospectors.

These numbers are dependent on the real number of plant species left to be screened, which is unknown at this point (Miller 2011). Still, this allows for a good number of new drugs, at least until obstacles in actual drug development are considered.

Mitigating factors could substantially lower predictions of undiscovered drugs. It is possible that many undiscovered secondary compounds will actually turn out to be compounds already identified (Balick & Cox 1996). Will the ratio of elucidated drugs from undiscovered secondary compounds be the same as those already identified?

Indigenous peoples have been inhabiting rainforests for thousands of years, and some researchers speculate that indigenous knowledge of medicinal plants is finite. If so, there could be a dramatic fall off of new drugs waiting to be identified (Principe 1991). Finally,

107 as the number of plant species screened increases, some believe that the number of undiscovered compounds could shrink disproportionately (Miller 2011).

Much of the uncertainty rests on how many new secondary compounds can be synthesized. Advancements in biotechnology may increase the number of compounds that can be extracted from plants. Or it might not. What is resolved is that bringing a drug to market from a plant is an expensive, herculean, and elusive task.

Still, tantalizing tales of a global pharmaceutical market worth $1 trillion in 2015

(Petrova 2014) create unrealistic expectations on benefit sharing agreements. This has caused a new problem. Rather than serving as an incentive, the prospect of benefit sharing can act to raise unrealistic hopes of monetary gains from medicinal plants. When this goes unrealized, indigenous peoples and host governments may feel cheated, even if the plant-to-drug research is a complete bust.

A bioprospecting incident in Cameroon serves as an example. A research team funded by the NCI was investigating a plant compound for anti-HIV bioactivity. The research was requiring significant investments with no success. Cameroonian government officials were becoming impatient at the lack of financial return. Ultimately, the plant did not pan out. But expectations remained. “The Cameroonians were all wanting to buy themselves new Mercedes,” said a person familiar with the research (Conniff 2012, 2).

Pharmaceutical Companies as the Good Guys: Two Case Examples

Most of the biopiracy narrative is focused on faceless Big Pharma conglomerates whose primary goal is profitability. But what if a company was founded upon the intention of putting benefit sharing with holders of indigenous knowledge and profits on

108 equal footing? Would the results be different? Two following case examples are not comforting for the future of bioprospecting benefit sharing arrangements.

The INBio-Merck Benefit Sharing Agreement: A Revolutionary Pact

Even before the CBD was inked in the early 1990s, attempts at revenue sharing between pharmaceutical companies and indigenous peoples were already underway. The most heralded was the INBio arrangement in Costa Rica. On October 24, 1989, the La

Asociacion Instituto Nacional de Biodiversidad (INBio) was established as a private, non-profit entity with the objective to inventory the country’s rich endemic flora and fauna. Considering Costa Rica’s vast biodiversity, this was no small task. Costa Rica is similar in size to West Virginia yet is home to approximately 5 percent of all known plant and animal species. The goal of INBio was to ensure the conservation of this vast biodiversity through commercial but sustainable uses (Hunter 1997). The initiative came just as interest in traditional medicine was experiencing a revival.

In late 1991, Merck & Co. Ltd., a well-known multi-national pharmaceutical firm, signed a 24-month deal with INBio that was heralded as a revolutionary bioprospecting agreement. In exchange for $1 million, plus a smaller up-front fee to pay for technical equipment, Merck would analyze Costa Rica’s rich plant biodiversity in hopes of finding chemicals that would become future pharmaceuticals. INBio was to receive an undisclosed percent of royalties (probably one to three percent) generated from any successful drugs (Aseby & Kempenaar 1995; Hunter 1997). The agreement seemed a model on which all future arrangements were to be based.

The deal was renewed in 1993 and again in 1996. Based on the biodiversity found in Costa Rica, it might have been expected that the program would return at least a

109 handful of previously unknown bioactive compounds for drug development. Yet not one marketable drug was ever brought to market. In 2008, Merck gave up its quest to find drugs from bioprospecting and gave away its inventory of 100,000 extracts from natural compounds (Conniff 2012). Thus ended one of the most anticipated bioprospecting arrangements in one of the most biologically diverse countries on the plant.

Shaman Pharmaceuticals: A Painful Reality

If there was ever a pharmaceutical company that seemed dedicated to the welfare of indigenous healers, it surely was Shaman Pharmaceuticals. Its founder Mark J. Plotkin became fascinated by traditional healing techniques while a university student. In the

1990s, Plotkin’s book about his encounters with traditional healers was widely read. The book’s success helped propel the renewed interest in bioprospecting based on TEK.

Understanding the suspicion with which many indigenous people viewed outside bioprospectors, and truly concerned about the future of indigenous peoples as modernity encroached, Plotkin decided to change that. He designed Shaman Pharmaceuticals with benefit sharing in mind. In addition, Shaman’s non-profit twin, the Healing Forest

Conservancy, was intended to use profits and donations to protect the tropical rainforests home to traditional healers and indigenous peoples (Brown 2003; Garrity & Hunter-

Cevera 1999).

The strategy of Shaman Pharmaceuticals was to forego the traditional shotgun route of mass plant screening. Instead, Shaman focused on cooperating with indigenous peoples to mine their millennia of traditional healing knowledge. Many were skeptical.

But the strategy looked like it was paying off. Shaman was launched just in time to tap into the rising tide of venture capital during the 1990s. Lisa Conte, the president and CEO

110 of Shaman Pharmaceuticals, told Business Week that Shaman’s success at finding bioactive leads was 50 percent from just 75 samples—a shocking success rate. By the mid-1990s, Shaman Pharmaceuticals was upbeat about two potential anti-viral drugs called Provir and Virend. Based on this early success the company formed partnerships with Merck and Eli Lilly (Brown 2003). It appeared Shaman Pharmaceuticals was about to do what had long eluded the pharmaceutical bioprospecting world: create a partnership with traditional healers that allowed drug companies to profit not at the expense of, but rather, in conjunction with traditional healers. Then the harsh reality of drug development came knocking.

By 1998, five years after the company’s initial public offering, Shaman

Pharmaceuticals found itself on shaky financial ground. Estimates put monthly capital expenditures at $4 million. The company made two subsequent stock offerings in an effort to stave off financial collapse while the Food and Drug Administration weighed whether Provir was market ready. When the FDA decided further testing was necessary

(due to a variety of factors), Shaman Pharmaceuticals’ ambitious goal collapsed (Brown

2003). In 1999, a decade after incorporation, Shaman Pharmaceuticals closed its doors

(Peterson 2001). Conte reinvented Shaman Pharmaceuticals as Shaman Botanicals, with the intent of concentrating on the exploding international natural-products market. By marketing its products as dietary supplements, Shaman Botanicals could avoid the strict

FDA scrutiny reserved for drugs (Brown 2003).

Shaman Pharmaceuticals’ problems did not end there. The company also fell victim to claims of biopiracy. These allegations arose out of two patents the company holds on sangre de drago (also sangre de grado). Some claimed that the patents, originally

111 for respiratory problems in children and another for herpes, are not unique due to uses of the plant for these very health issues over the millennia. With Shaman Pharmaceuticals’ bankruptcy, a probable result is that the patents will linger in limbo until falling into the public domain. The patents will neither provide benefit sharing to indigenous peoples, nor provide the world at large with needed drugs (Reid 2010).

Shaman was also plagued by the same dilemma other bioprospecting arrangements have been faced with: how to draw boundaries between those groups or communities who participate in benefit sharing and those who do not. Despite distributing upwards of $3.5 million to indigenous groups in various countries, Shaman

Pharmaceuticals and the Healing Forest Conservancy came under fire for not sharing with the correct groups.

It has already been established that plants used in traditional healing are not confined geographically to any arbitrary border drawn on a map. Rather, medicinal plants are often found over a wide geographic domain where different groups and cultures access the plants, sometimes for the same purpose. Thus, Shaman Pharmaceuticals and the Healing Forest Conservancy were forced to decide if benefit sharing would take place only with groups with which the organizations had collaborated, even though this could cut out other groups located very close geographically (or culturally) (Brown 2003). Even with the best intentions, no matter how lines are drawn, one group will always be deemed the winner and another the loser.

Shaman Pharmaceuticals started with a noble ambition but fell victim to the reality of the incredibly hard, expensive, and torturously long process needed to bring a successful drug to market. This reality is often overlooked in biopiracy rhetoric.

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Chapter Summary

Despite high hopes, the post-CBD bioprospecting and benefit sharing agreements did not generate vast wealth for holders of traditional knowledge. It did promote unrealistic expectations of economic wealth based on biological resources (Brush 2004).

Almost ten years after the CBD was adopted, no commercialized product had been developed by the pharmaceutical industry based on TMK (Moran, King, & Carlson

2001). As the following chapters will show, by this time there was a noticeable expansion into microbial bioprospecting.

The initial enthusiasm for combinatory chemistry also subsided. As late as 2012, only one new chemical entity (NCE) that was identified through combinatory chemistry had become a marketable drug. That was sorafenib, under the name Nexavar, used to treat a form of carcinoma (Newman & Cragg 2012). Yet combinatory chemistry or some form of advanced biotechnology is almost always required to create an effective drug from a plant compound. The cost and difficulty of bringing a drug to market from a plant is so difficult that “ . . . only in extremely rare instances, does the isolated natural product itself serve as a magic bullet” (Cordell 2000, 469).

This chapter demonstrated how the illusion of the easy plant-to-drug narrative is false. The chapter showed using traditional ethnobotanical knowledge as a direct path to drug discovery is problematic, and that even a pharmaceutical company founded on the principle of benefit sharing was unable to turn a profit. Yet these beliefs—that creating pharmaceuticals from drugs is easy, creates instant wealth, and that indigenous knowledge is key to this process—were significant drivers in shifting bioprospecting toward the world of microbial organisms.

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CHAPTER NINE

BIOPROSPECTING IN AFRICA

An Argument for Contemporary Biopiracy

The example of quinine derived from the cinchona tree was used earlier to demonstrate how an actual act of biopiracy served to fuel suspicion of researchers in the present age. More contemporary examples often cite the rosy periwinkle. Yet that has been shown to be a dubious example of biopiracy in the modern age. To dismiss all claims of biopiracy as mere patent disputes or overly suspicious minds is as equally troubling as those who view bioprospecting as little more than sanctioned biopiracy.

Is there a contemporary case that might stand as a good example of biopiracy in recent decades? In fact, there is. It comes from Africa. Not from the equatorial rainforest nor the country of Madagascar, but instead the arid expanse of the Kalahari Desert in southern Africa.

Hoodia: A Case Study

Hoodia gordonii, known generically as Hoodia, would be dismissed by most as an average succulent. It belongs to the Apocynaceae family. It thrives in hot, arid environments of South Africa and neighboring Namibia. The first written description of the plant by Europeans was by the botanist Carl Peter Thunberg in 1773 (Balick 2007;

Vermaak, Hamman, & Viljoen 2011). In the past decade Hoodia became internationally known for two reasons: a potential diet drug and a strong case for biopiracy.

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The San are considered to be the original inhabitants of southern Africa. They have practiced a nomadic lifestyle in the Kalahari Desert for at least 20,000 years. About

2,000 years ago, a pastoralist people called the Khoi migrated into the area. They were followed by the Xhosa about 1,400 CE (Chennells 2013). See Figure 11.

Figure 11. Yellow boundary represents approximate extent of Hoodia gordonii. Adapted from Swart, 2008; Google Earth, 2013. Inset of Hoodia gordonii in Namibia. Van Rooyen, 2001, Kew Royal Botanical Gardens website. African countries shapefile by Flannery, 2014.

The San have may have the dubious reputation as being the most marginalized of all of the groups in southern Africa. Colonization led to a loss of land plus a fractured culture and identity (Chennells 2007). In the early 1890s, the San clashed with the

German settlers in the region. San women were raped. Retaliation against white farms by the San resulted in a state-sanctioned policy of genocide (Osseo-Asare 2014). The San are now dispersed, with about 9,000 residing in South Africa, 35,000 in Namibia, and

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55,000 in Botswana. There are smaller populations in Angola, Zimbabwe, and Zambia

(Chennells 2013, 2007). Enduring poverty appears to be the one factor that all San share

(Vermaak, Hamman, & Viljoen 2011).

To ward off pangs of hunger and to gain moisture, the San commonly consume

Hoodia, which is similar in size to a cucumber. The San are generally regarded as the originators of the knowledge that Hoodia was an effective appetite suppressant (Balick

2007; Chennells 2004). Many species of Hoodia are generically referred to as “ghaap.”

Since some species have different uses, Hoodia gordonii is sometimes credited for medicinal remedies against stomach cramps and hypertension (Vermaak, Hamman, &

Viljoen 2011). It is now known that secondary compounds in Hoodia gordonii called glycosides function as appetite suppressors (van Heerden 2008).

Researchers over the past 100 years have been baffled as to how the San are able to survive in a geographic region that offers little water or food. In the 1960s, over 1,000 regional plant species were analyzed by the South African Council for Scientific and

Industrial Research (CSIR) for possible diet or drug potential. Plants used by indigenous communities were targeted. Samples of plants regularly consumed by the San were obtained, including Hoodia gordonii and Hoodia pilifera (van Heerden 2008; van

Heerden, Horak, Maharaj, Vleggaar, Sanabe et al. 2007).

However, research into Hoodia stalled. In the 1980s, armed with new technology, researchers again examined Hoodia, but this time for weight loss possibilities. By the mid-1990s, researchers had isolated a bioactive chemical in Hoodia that made it desirable as an appetite suppressant (Chennells 2007; Osseo-Asare 2014). In 1996, a patent was

116 awarded to CSIR for a chemical named P57. A year later, CSIR signed a deal with a natural products company in Britain called Phytopharm.

Where exactly the idea came from to investigate Hoodia as a hunger suppressant is unknown. Starting in the 1700s, the Hoodia plant served not only as a food source for the San, but also Dutch colonists (Osseo-Asare 2014). Chennells (2004), a counsel at the

South African San Institute, pointed to a Dutch ethnobiologist who authored a 1937 paper on Hoodia, or the South African military’s San trackers, as likely sources that brought the plant’s properties to the attention of the CSIR. Although the extent of interaction between the Xhosa and the San is unclear, there is evidence that the Xhosa absorbed a good deal of the medicinal knowledge practiced by San healers (Chennells 2013).

The Council for Scientific and Industrial Research records do not reveal the names of the Khoi-San who may have provided knowledge of the plant’s properties.

Even the exact geographic location is unknown. The Hoodia plant grows not just in parts of South Africa, but in Namibia and Botswana as well. The plant had obviously come from some region traditionally inhabited by San groups (Osseo-Asare 2014; Vermaak,

Hamman, & Viljoen 2011).

In 1998, Phytopharm signed a sub-licensing deal with pharmacutical manufacturer Pfizer, Inc. (Chennells 2013). All of this proceeded without anyone consulting the San. That changed in 2001, when Hoodia’s potential as an appetite suppresant drug gained global attention (Vermaak, Hamman, & Viljoen 2011). It was soon asked why a people who had long used a plant in a specific geographic area were not receiving compensation (Osseo-Asare 2014). The head of Phytopharm reportedly told

117 a British paper that the San were “extinct” (Chennells 2007, 13). This was contradicted by the approximately 100,000 San living in southern Africa (Chennells 2004).

The South African San Council filed a lawsuit. In 2002, CSIR acknowledged that

San traditional knowledge had provided the initial lead. In 2003, the San Council accepted an agreement with CSIR that would allow the San groups to share in revenue from Hoodia-related products. Other San communities residing in Namibia and Botswana pressed for a share of revenue and eventually received it (Chennells 2007). Around the same time, South Africa adopted its Biodiversity Act to keep it in compliance with the

Convention on Biological Diversity (Osseo-Asare 2014).

The San seemed poised to be one of the first real breakouts for benefit sharing envisioned by the CBD. The international Hoodia market took off. All sorts of dietary supplements supposedly derived from the plant hit the shelves. It is doubtful some of the products even contained any Hoodia extract. Farmers raced to plant enough Hoodia.

There were concerns of the plant going extinct in some places due to unauthorized harvesting (Chennells 2007). The supposed dietary wonder drug even got a spot on “60

Minutes” (Osseo-Asare 2014).

Then things started to go downhill. Uncertainties ranging from inability to secure enough plant material, to high development costs, to safety concerns, had stalled development by 2008 (Osseo-Asare 2014; Vermaak, Hamman, & Viljoen 2011).

Phytopharm eventually returned development of a diet drug from Hoodia back to CSIR

(Osseo-Asare 2014). Despite the San having received a favorable deal of six percent on royalties and eight percent on milestone payments, no commercially successful products

118 have been generated from Hoodia (Vermaak, Hamman, & Viljoen 2011). The San became the victims of the incredibly hard task of bringing a drug to market.

Hoodia v. the Rosy Periwinkle

There are several important differences between Hoodia and the rosy periwinkle.

The San succeeded in their claims because they were a distinct group in a distinct region where Hoodia is native. Hoodia is difficult to transplant, so the plant does not grow elsewhere. Therefore, local people have harvested only from this one area (Osseo-Asare

2014). The rosy periwinkle grows in numerous locations and has been used by many groups over a wide geographic area. Unlike the rosy periwinkle, Hoodia’s traditional usage as an appetite suppressant, and the original investigation into the plant for that purpose, did result in an appetite suppressant drug (although short-lived). The rosy periwinkle was investigated as a diabetic remedy, and instead became an anti-cancer drug. Finally, the end of apartheid and changing politics made CSIR researchers acknowledge that traditional knowledge of Hoodia had in fact been utilized in research without the consent of the San, and that those findings had been kept secret from the San

(Osseo-Asare 2014).

Still, Hoodia raises questions on other levels. As Chennells (2007) observed, the

San provided their consent only after research had been conducted. It raises the question that at what point in time does prior informed consent become prior. Chennells further points out that the San traditionally do not possess a formal leadership that speaks for everyone. Yet the San were represented by a council when negotiations took place. This raises issues similar to the Maya ICBG in regards to deciding what entity can speak for the entire community.

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Chapter Summary

The Nagoya Protocol was not in effect during the Hoodia example discussed. It is hard to say how helpful it might have been in negotiations for the San. The San seemed to have everything going for them: a fairly clear-cut case of a marginalized people possessing a relatively geographically-contained knowledge of medicinal plant usage; a plant that came from a defined geographic region; an extract poised to become a major drug, and; a fair benefit sharing deal. But it all fell apart.

The previous chapters have discussed bioprospecting with plants as the focus. The following chapters will explore bioprospecting’s new era. Although plants are not at the center of this endeavor, the expansion into microbial organisms may ultimately create equally contentious issues.

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CHAPTER TEN

A NEW ERA IN BIOPROSPECTING

Microbial Bioprospecting

Chapter Introduction

The lure of finding the next miracle drug has not changed. What has changed is the landscape upon which bioprospecting is being conducted. Rainforests have given way to fumaroles. Steamy tropics have given way to Antarctic ice. Terra firma has given way to the marine abyss. Bioprospecting has expanded into a new era that focuses on organisms too small to be seen with the unaided eye. Steaming fumaroles, the deep marine abyss, and the frigid Antarctic are all the next frontiers of bioprospecting.

Microbial Organisms: Endless Possibilities

When the first Europeans landed in the tropical rainforests of the Americas they found a cornucopia of bizarre plants. As bioprospectors peel back the microbial world, they are finding organisms even more fascinating, and potentially more useful.

Regardless of how extreme of an environment, microbes appear to be able to withstand the conditions (Cavicchioli 2002; van den Burg 2003).

Microorganisms can be found in the domains of Bacteria, Archaea, and Eukarya.

Microorganisms do not have a strict definition. Generally, they can be thought of as organisms whose mass does not exceed 10-5 grams and possess a length no more than 500

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µm (Martiny, Bohannan, Brown, Colwell & Fuhrman et al. 2006). This microscopic life is 50 times tinier than a human hair follicle (Yellowstone Association 2015).

Microbial bioprospecting began generating a great deal of interest in the mid-

1990s. This is not attributable to a specific reason. Rather, bioprospecting’s expansion into the realm of microorganisms appears to have been a convergence of several factors.

Possibly, some researchers and bioprospectors were frustrated by the vague guidelines and perceived inadequacies of the CBD, as discussed earlier. This may have prompted bioprospectors to look elsewhere for sources of secondary compounds.

Advancements in biotechnology certainly were a factor. Akondi and Lakshmi

(2013) credit a robust improvement in molecular tools around 1995 as a major development in the ability to study microbial organisms. Increased capability with PCR

(polymerase chain reaction) technology made it possible to screen previously uncultured microbial organisms (Bull 2004). High-throughput screening using culture-independent methods began to accumulate large metagenomic libraries that in turn could be analyzed to identify more bioactive agents (Akondi and Lakshmi 2013).

In the mid-1990s, only one percent to five percent of microbial species diversity had been identified (Hunter-Cevera 1998). With the advent of new technology, researchers were able to begin tapping this immense reservoir of biodiversity. Still, the vastness of microbial life is so great that the estimate of one percent of microbial life having been explored remains unchanged (Otohinoyi & Omodele 2015).

In 1997, the National Park Service announced a benefit sharing deal between

Yellowstone National Park and a private biotech company (Yellowstone National Park

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2015). This increasing interest in microbial organisms left little doubt that bioprospecting appeared to be entering a new era.

Microorganisms are far and away the largest group of life on the planet. Microbes form 60 percent of the planet’s total biomass (Dionisi, Lozada, & Olivera, 2012).

Knowledge of the microbial world is limited, but the biodiversity of that world is suspected to be vast. Some estimate that microbial organisms account for at least 90 percent of the world’s diversity (Schafer & Borchert 2004). A recent application of scaling laws applied to microbial organisms found there is approximately 1012 (one trillion) microbial species on Earth (Locey & Lennon 2016).

Microorganisms have been evolving on this planet for 3.5 billion years. They have adapted to environments once thought impossible for life (Schafer & Borchert

2004). Microbes have been discovered deep within the bowels of the Earth where they thrive, sometimes in a dormant state. There they survive by energy obtained from the hydrogen or carbon dioxide provided by geochemical reactions within rocks (Hunter-

Cevera 1998; Raven & Johnson n.d.). Even deserts may contain microbes, which can be active or dormant for long periods (Rothschild & Mancinelli 2001).

Plants contain a good number of microbes called endophytes (Strobel & Daisy

2003). Soil also has a high number of microbes (Synnes 2007). This thesis focuses on microbes that are found in polar climates, the marine abyss, and fumaroles on land.

Microbial organisms have already been used in a variety of applications. A group of microbes called extremophiles possesses potential for a wide array of applications, from pharmaceutical to industrial (Otohinoyi & Omodele 2015). Uses in agriculture include nitrogen fixation, herbicides, and pesticides (Hunter-Cevera 1998: Krishnamurthy

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2003). Microbial extremophiles have demonstrated potential in biomining (the use of biological processes to extract ores), ore-bleaching, and de-sulfurization of coal. Other industrial applications are in detergents; cheese and dairy production; food sweeteners and flavoring agents; and baking and brewing. Microbial organisms can be used for environmental purposes like bioremediation and waste disposal (Dermijian, Moris-Varas

& Cassidy 2001; Hunter-Cevera 1998; Krishnamurthy 2003). Biotechnology includes uses for genetic engineering (Dermijian, Moris-Varas & Cassidy 2001). More exotic uses of microbes include applications in producing solar energy (Krishnamurthy 2003).

Pharmaceutical applications are no less varied. Typhoid and Hepatitis B vaccines are extracts from microbial enzymes. Trans-genetically altered microbes have been used in steroids and antibiotics. Microbial organisms provide a host of useful enzymes in medicines (Krishnamurthy 2003). See Table 1 for a list of applications for antibiotics derived from microbial organisms.

The Extreme Worlds of Extremophiles

A particular type of microbe is of the greatest interest to bioprospectors.

Extremophiles are a microorganism that survives, or even thrives, in conditions unbearable to humans (Cavicchioli 2002). Extremophiles can be thought of as any microorganism that thrives in any environment thought too extreme for life (Rothschild

& Mancinelli 2001). Extremophiles favor niches that have extremely high or extremely low temperatures (fumaroles and ice sheets); high or low acidity; crushing pressures in the ocean depths; extremely high salinity (NaCl); or even radiation.

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Table 1. A Partial List of Pharmaceutical and Industrial Uses of Antibiotics Derived from Microbial Organisms

Useful Antibiotics Derived from Microbes

Microbial Antibiotic Current Applications

Cephalosporins, Penicillins, Rifamycins antibacterials

Amphotericin B., Griseofulvin, Nystatin, Hamycin antifungal

Abikoviromycin, Kikumycin antiviral

Adriamycin, Belomycin antitumor

Avermectin, Hygromycin anthelminithic

Herbicidin herbicide

Mibemycin miticide (insecticide)

Tectranectin insecticide

Gibberellins plant hormone (agriculture)

Monascin food pigment

Detoxin detoxicant

Azalomycin antiprotozoal

Nissin food additive

Variginiamycin animal growth promoter

Colisan, Patulin antispasmodic

Cyclosporin A., Alanosine immunosuppressive

Griseofulvin, Amicomycin anti-inflammatory

Dopastin, Spiramycin hypertensive

Filipin, Nogalomycin anticoagulant Note: Adapted from Krishnamurthy, 2003.

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The environment of microbes known as extremophiles is believed to be limited only by temperature (Hunter-Cevera 1998). Even the range of temperatures—from gut- wrenching cold to boiling hot—that extremophiles endure is phenomenal (Schafer &

Borchert 2004; Srivastava, Rai, Kumar, Kashyap, & Arora 2013). Microbes of the

Archaea family are of particular interest for researchers due to their ability to thrive in a combination of extreme temperature, salinity, and acidity, plus overall toxicity (Synnes

2007). It should be noted that extremophiles can also be eukaryotes (multi-celled organisms), although most thermophiles are prokaryotes. This is because eukaryotes are unable to survive at temperatures higher than ~60 ºC (140 ºF) (Rothschild & Mancinelli

2001).

Extremophiles are able to withstand the brutal conditions of extreme environments through evolution (Hunter-Cevera 1998; Schafer & Borchert 2004). A cell’s lipids, proteins, and nucleic acids (known as biomolecules) all must adapt to life under punishing conditions (Rothschild & Mancinelli 2001; Synnes 2007). Since complex DNA is vulnerable to extreme temperatures of >60º C (76º F), extremophiles in high temperature environments have DNA sequences that can tolerate extreme temperatures (Raven & Johnson n.d).

Thermophiles and hyperthermophiles flourish in high temperatures because of a compact and durable monolayer cellular membrane that resists heat far more effectively than the traditional bi-layer cell membrane (Raven & Johnson n.d). Uniquely adapted enzymes allow a microbe to survive in supracold environments. These allow a microbe to tolerate cryptobiosis, a period of dormancy brought on by extremely low temperatures.

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Or, enable the cell to produce proteins (cryoprotectors) that withstand freezing

(Srivastava, Rai, Kumar, Kashyap, & Arora 2013).

Scientists believe prokaryotes inhabit these niches because they are too extreme for multi-celled organisms (Srivastava, Rai, Kumar, Kashyap, & Arora 2013). In the process, extremophiles produce a diverse number of enzymes that may have numerous potential applications in industry and medicine (Synnes 2007). The reason for this amazing versatility is that extremophilic organisms give rise to biomolecules that are much more stable in extreme conditions (temperature, pressure, salinity, etc.) than enzymes found in mesophilic organisms (organisms that tolerate moderate temperature ranges) (Otohinoyi & Omodele 2015).

Types of Extremophiles

Extremophiles are so diverse in their ecological niches that various names have been given to them, based on the extreme environments which they inhabit.

Extremophiles classified by temperature are either thermophiles (punishing heat), or psychrophiles (withstanding brutal cold, such as polar ice or ocean depths). Acidophiles and alkaliphiles can tolerate various degrees of pH. Xerophiles thrive in extreme arid niches, like deserts (hot or polar). Microbes that survive buried deep within rocks are endophiles (endoliths). Halophiles tolerate high sodium environments. Extremophile environments can be extremely acidic (up to 10 percent hydrochloric acid), highly alkaline, or have high concentrations of sodium (Johnston & Lohan 2005; Rothschild &

Mancinelli 2001; Srivastava, Rai, Kumar, Kashyap, & Arora 2013; Synnes 2007; Vorgias

& Antrankian 2004). Add in the crushing pressure of the benthic abyss and these extremophiles get even more amazing.

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One certain type of extremophile generating a lot of interest is known as a thermophile. These microbes inhabit brutally hot environments. So great is their diversity of environments that there are grades of thermophiles. They generally exist in environments over 50 ºC (122 ºF). Those existing in temperatures of 50-60 ºC (122-140

ºF) are moderate thermophiles. Extreme thermophiles grow in 60-80 ºC (140-176 ºF).

Some environments are so extremely hot that the term thermophile is not satisfactory.

Instead, a second class has been added to describe microbes that thrive in superheated environments—hyperthermophile. Hyperthermophiles tolerate temperatures of 80 ºC –

112.8 ºC (176-235 ºF) (Srivastava, Rai, Kumar, Kashyap, & Arora 2013; Synnes 2007;

Vorgias & Antrankian 2004).

These searing environments favored by extremophiles and hyperthermophiles are found in fumaroles where superheated water or sludge is disgorged by vents in the

Earth’s surface, such as those in Yellowstone National Park. At places like the Mid-

Atlantic Ridge, over 12,000 feet deep, water from hydrothermal vents called black smokers can reach temperatures of 400 ºC (752 ºF). Black smokers, vents on the ocean floor pouring out scalding hot water in addition to sulphur or a mixture of other otherwise deadly chemicals, are favored by thermophiles. Enzymes from these microbes can be used in a variety of high-temperature energy applications (Srivastava, Rai, Kumar,

Kashyap, & Arora 2013; Synnes 2007; Vorgias & Antrankian 2004).

Psychrophiles, microbes favoring punishingly cold temperatures, are also divided into different categories. Psychrophilic microbes thrive in permanently cold environments of 15 ºC (59 ºF) and lower. None have been found at temperatures higher than 68 ºF

(Reed 2005; Srivastava, Rai, Kumar, Kashyap, & Arora 2013). Psychrotrophic microbes

128 prefer seasonally cold environments. Both grow best when temperatures hover close to 0

ºC (14 ºF) or lower. These environments generally are found in the Antarctic and Arctic ocean regions that remain frozen a majority of the time. The natural anti-freeze of these organisms has various industrial and medical applications. Psychrophiles are used in a variety of applications from cold laundry detergents (making cold-water detergents more effective, thus saving energy), wine making, and cheese manufacturing (Srivastava, Rai,

Kumar, Kashyap, & Arora 2013; Vorgias & Antrankian 2004).

Acidophiles are those microbes that thrive in a pH range of 5.0 to 0.7 (with pH

3.0 being optimal), generally in temperatures of 60 to 90 ºC (140 to 194 ºF). This makes acidophiles potentially useful in coal and ore mining and production. Alkaliphiles do best in environments above a pH 9, but have been found in East African lakes with a pH of

12.0. Haloalkaliphiles combine high tolerances for alkaline with high tolerances for sodium. Carbonate-rich springs and lakes are a prime environment. These extremophiles have various industrial applications such as degreasers (Srivastava, Rai, Kumar, Kashyap,

& Arora 2013; Vorgias & Antrankian 2004).

Halophiles are possibly the most diverse group of extremophiles. They have a powerful resistance to the osmotic pressures of salt-rich environments. Halophiles do best in three to 30 percent saline environments. Possible industry-related applications include resistance to freezing or desiccation, or enduring high temperatures (Reed 2005;

Srivastava, Rai, Kumar, Kashyap, & Arora 2013; Vorgias & Antrankian 2004).

Piezophiles were formerly called barophiles. Although some do well at surface atmospheric pressure, most prefer pressures reaching 40 MPa and above (equivalent to

394 atmospheric pressures at sea level). Possible industrial applications involve food

129 production where high pressures are required to sterilize and process food. Due to the difficulty of reproducing extreme pressures in the lab, scientists are still pondering the potential applications for piezophiles (Parkes & Wellsbury 2004; Srivastava, Rai, Kumar,

Kashyap, & Arora 2013)

Xerophiles thrive in conditions of limited of water. Xerophiles are responsible for spoiling food. Both mold and yeast are well-known xerophiles. Xerophiles have the ability to thrive in low water and low humidity environments, which makes them potential targets for water-saving applications. Xerophiles in the Antarctic can survive in long periods of darkness (Rothschild & Mancinelli 2001; Srivastava, Rai, Kumar,

Kashyap, & Arora 2013).

Radiophiles, also known as radiation-resistant extremophiles, are microbes that can withstand radiation that would be lethal to other organisms. Some have even been found living in the cores of nuclear reactors (Cavicchioli 2002). These microbes are also found in places of extreme elevation that receive a great deal of ultra-violet light.

Radiation-resistant extremophiles are of particular interest because of their potential to repair DNA, plus resistance to oxygen depletion and lack of nourishment. This makes them candidates for anticancer drugs, along with industrial uses in agriculture (Demirjian,

Moris-Varas, & Cassidy 2001; Gabani & Singh 2013;).

Microaerophiles, extremophiles that survive in depleted oxygen environments, also offer possibilities in a range of applications (van den Burg 2003). Extremophiles can also come in many combinations of the above. These microorganisms are called polyextremophiles (Otohinoyi & Omodele 2015). See Table 2 for extremophiles and their environments.

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Table 2. Habitats of Extremophiles

Type Habitat General Optimal Environment Environment

Thermophile Fumaroles; Deep sea vents; Heat tolerant 113˚F - 176˚F Hot springs; Geysers

Hyperthermophile Fumaroles; Deep sea vents Extreme heat tolerant 130˚F - 235˚F

Psychrotolerant Frigid water; icy zones Cold tolerant 39˚F - 46˚F

Psychrophiles Frigid water; Arctic & Cold tolerant <32˚F - 53˚F Antarctic ice

Acidophiles Hydrothermal vents; hot Extreme acidity <0.7 pH to 3.0 springs; acidic solutions; pH mine drainage

Alkaliphiles Carbonate or Soda lakes; Extreme alkaline 9.0 pH to 13.0 highly alkaline solutions pH

Halophiles Hypersaline lakes (Dead Extreme saline 25% to 32% Sea, Great Salt Lake) solutions NaCl

Barophiles / Abyssal depths Intense pressure > 1 to >500 Piezophiles atmospheres

Radiophiles Elevated altitudes Elevated radiation High UV; (mountains); high UV levels gamma rays environments (open fields); nuclear power plants

Microaerophilic Oxygen-deprived lakes; Low oxygen levels <21.0 % O2 hypoxic areas

Metalophile Mining sites; geologic Elevated metal zones concentration

Xerophiles Desiccated environments Deserts; Antarctic Extremely arid deserts Note: Extremophiles take advantage of nearly all environments and as a result offer a cornucopia of pharmaceutical and industrial possibilities. Extremophiles can occur in combinations, such as an acid-tolerant thermophile. Compiled from Raven & Johnson, n.d.; Gabani & Singh, 2013; van den Burg, 2003; Rothschild and Mancinelli, 2001; Dermijian, Moris-Varas, Cassidy, 2001.

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Obtaining useful enzymes from extremophiles consists of two alternative major steps. First, the necessary biomass must be produced from the enzyme to extract and analyze the biocatalytic substances (enzymes that modify a chemical reaction). Or, the biocatalyst gene codes can be cloned and replicated in a host. Both of these methods require expensive and cutting-edge technology that is rapidly evolving (van den Burg

2003).

Although there are currently over 3,000 enzymes used in a range of applications, many are unstable in extreme conditions and processes. Enzymes or “extremozymes” from extremophiles can withstand these processes. The biocatalysts (enzymes that facilitate chemical reactions) extracted from extremophiles remain stable in such conditions. Additionally, it is suspected that the genomic sequences of extremophiles may in turn produce unique enzymes for innovative applications (Kumar, Aswathi, &

Singh 2011).

Chapter Summary

Since the mid-1990s, bioprospecting has steadily expanded into a new era of microbial bioprospecting. Microbes known as extremophiles are of particular interest.

This expansion, or new era of bioprospecting, was brought on by a number of events that reached a denouement during the 1990s. These were discussed in previous chapters. An additional factor stemmed from bureaucratic hurdles following implementation of the

CBD that created frustration and appeared to be stalling some plant-based research projects.

Most importantly, innovations in biotechnology appeared in the 1990s that allowed scientists to isolate enzymes in microorganisms that had previously eluded them.

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The elucidation of extremozymes isolated from microorganisms had a profound impact.

Since the evolution of these extremophiles enabled them to exist in places where life was thought impossible, it was realized that their enzymes could be used in a wide array of industries and processes where previously known enzymes were unsuitable.

Further, many of these extremophiles are found in areas almost off the map, in deep ocean vents or the frigid interior of Antarctica. One very inviting spot for extremophile bioprospecting does not reside off the map. It is currently taking place in the United States. The next chapter examines how America’s national parks played, and are continuing to play, a critical role in the expansion of microbial bioprospecting.

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CHAPTER 11

MICROBIAL BIOPROSPECTING IN AMERICA’S NATIONAL PARKS

A New Bioprospecting Frontier

Chapter Introduction

The expansion into microbial bioprospecting in the mid-1990s enlarged the geography of bioprospecting. Most Americans would probably be surprised to know that for almost two decades commercial bioprospecting has been taking place on a regular basis in America’s national parks. One park in particular is drawing the most microbial bioprospectors.

Yellowstone National Park (YNP) contains over 10,000 assorted hydrothermal features including fumaroles, mud pots, and steam vents (Smith & Siegel 2000; Varley &

Scott 1998; Yellowstone National Park 2015). The extremophiles living in these harsh thermal environments may rival the biodiversity found in tropical rainforests (Wood

2000). These extremozymes could offer a wide array of pharmaceutical and industrial applications (Varley & Scott 1998). Potentially, the NPS could reap sizable revenues from commercial applications resulting from Yellowstone’s microbes. In 1997, the

National Park Service announced it had taken a major step in that direction by signing a benefit sharing agreement with a private company (Varley & Scott 1998; Yellowstone

National Park 2015).

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This chapter examines why Yellowstone National Park has captured the interest of microbial bioprospectors. The chapter will discuss microbial bioprospecting’s potential impact on America’s national parks. Can commercial arrangements be balanced with the public enjoyment and biological preservation of America’s parks? Or will bioprospecting in America’s national parks jeopardize the great biodiversity these parks contain? Once before a microbe that revolutionized biotechnology was plucked from the thermal pools of YNP. As bioprospecting expands into the microbial era, YNP could once again find itself the focus of the microbial bioprospecting world.

Bioprospecting for Hot Gold in the World’s First National Park

President Ulysses Grant signed the legislation creating Yellowstone National Park on March 1, 1872. There was little indication that reserving a parcel of land in the

Wyoming, Montana, and Idaho frontier (ten Kate, Touche, & Collis 1998; Wood 2000) would become one of humanity’s nobler accomplishments. Yet that stroke of pen launched America, and later the world, on a course of preserving great tracts of natural beauty and biodiversity. In a little over 100 years it would also create a biotechnology revolution.

When the NPS signed a benefit sharing agreement between Yellowstone National

Park and Diversa Corporation in 1997, it could be said that microbial bioprospecting had come full circle. For it was the discovery of the bacterial species Thermus aquaticus that led to the renewed interest in microbial bioprospecting. In 1966, Dr. Thomas Brock from

Indiana University was conducting research into thermophiles as part of a wider investigation into the evolution of life. A water sample from a thermal vent in

Yellowstone brought little attention at the time, other than that a microorganism had been

135 discovered thriving in water that was 71.1 ºC (160 ºF)—a feat previously thought impossible (Biel 2004; Mattix 1999). See Figure 12.

Figure 12: The pool that started it all. It was from this thermal pool called Mushroom Spring in YNP from which Thomas Brock plucked the organism known as Thermus aquaticus. Taq polymerase was derived from the organism, opening the door to new bioengineering possibilities for DNA. Rothschild & Mancinelli, 2001.

Samples of Thermus aquaticus collected by Brock were placed in the American

Type Culture Collection (ATCC) in Maryland for the purpose of public access (Bryson &

Kaczmarek 2000). A sample was obtained by the Cetus Corporation for $35.00 (ten Kate,

Touche, & Collis 1998). Twenty years after Thermus aquaticus was plucked from a thermal pool, an enzyme extracted from the microbe led to the development of the Taq polymerase process, known as the polymerase chain reaction (PCR). This scientific advancement accomplished two things. It garnered the Nobel Prize in Chemistry for Kary

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Mullis who directed the research at Cetus Corporation (Biel 2004; Doremus 1999), and it set forth a revolution in deoxyribonucleic acid (DNA) research.

The PCR process allowed previously nonculturable microbes to be reproduced in the laboratory by easily and efficiently reproducing small bits of complex DNA strands.

Doing so requires alternating cycles of extreme high and low temperatures. The Taq polymerase isolated from Thermus aquaticus was the only substance that could withstand the extreme heat required for the process. By using this process, billions of identical copies of DNA can be reproduced within hours. From a single microscopic fragment researchers can then produce enough material to study a substance in the laboratory. Its applications range from biotechnology, pharmaceuticals, and forensics (Biel 2004;

Bryson & Kaczmarek 2000; Mattix, 1999; ten Kate, Touche, & Collis 1998). Many television viewers are familiar with offshoots of this DNA fingerprinting process from shows like CSI: Crime Scene Investigation.

Following the development of the PCR technique by Mullis, F. Hoffmann-

LaRoche, a Swiss pharmaceutical company, purchased the patent to the PCR process for

$300 million. By the end of the 1990s, revenues associated with this process were estimated at over $1 billion annually (Biel 2004; Mattix 1999). Yellowstone National

Park, home to the thermal pool where Thermus aquaticus was discovered, has to date received . . . nothing. That is because at the time of Brock’s research, no revenue sharing agreements were in place. Brock had permission to remove samples, but was not legally required to share with the NPS any profits that might be derived from the research or biological material (ten Kate, Touche, & Collis 1998). It is a lesson from which the NPS took astute instruction. In 1997, the NPS signed its first ever benefit sharing agreement

137 with a biotechnology company called Diversa Corporation (now known as Verenium

Corporation) (Mattix 1999; Wood 2000; Yellowstone National Park 2015).

Bioprospecting had entered a new era.

The Diversa Corporation benefit sharing agreement came on the 125th anniversary of the creation of Yellowstone National Park (Doremus, 1999; Varley & Scott 1998;

Wood 2000). The agreement with Diversa Corporation was finalized in 1998. Officially known as a Cooperative Research and Development Agreement (CRADA), it granted

Diversa a “non-exclusive right to bioprospect” in the environs of Yellowstone National

Park within the scope of collecting “biological tissues, soils, sediments, water, and rocks”

(Wood 2000, 204). The CRADA is alternatively referred to as a revenue or profit sharing agreement. Any revenues generated from commercial applications derived from collected samples were to be shared with the NPS (Bryson & Kaczmarek 2000; ten Kate, Touche,

& Collis 1998; Wood 2000). It was heralded as a new day for America’s national parks.

From that day forward, if revenues like the one from Thermus aquaticus were realized, the NPS would share in the royalties.

There appears to be plenty of opportunity to discover the next Thermus aquaticus.

Yellowstone has a near monopoly on the world’s thermal features. The park contains 300 active geysers, which is 55 to 80 percent of active geysers in the world. Sixty percent of all geothermal features like superheated pools and fumaroles are located within the park

(National Park Service 2015; Smith & Siegel 2000; ten Kate, Touche, & Collis 1998).

The next largest collection of geysers is found on the Kamchatka Peninsula. The Dolina

Geizerov field has a mere 200 active geysers. New Zealand checks in with the third most geysers at 40, followed by Chile with 38. Fiery Iceland, probably thought to hold the

138 most geysers in the public mind, has only 25 active geysers (Smith & Siegel 2000).

Additionally, 60 percent of all geothermal features like superheated pools and fumaroles are located within YNP (National Park Service 2015; Smith & Siegel 2000; ten Kate,

Touche, & Collis 1998).

Barely one percent of the microbes thriving in Yellowstone’s thermal pools and associated environments has been identified and analyzed (Varley & Scott 1998). Since microorganisms compose the greatest molecular and chemical variety, it leaves open the potential for biotechnology breakthroughs. Microbial biodiversity is also enhanced by the

“island effect” of hot springs, of which YNP has a great number (Valverde, Tuffin, &

Cowan 2012). Possibly, the microbial biodiversity of Yellowstone’s thermal features could rival the diversity of tropical rainforests (Wood 2000). Whether the vast number of remaining unknown thermophiles will lead to a commercial pharmaceutical success is yet to be seen. Such high hopes for the potential of microbes is reminiscent of the earlier anticipation over new medicines that rainforest plants were expected to yield.

Although glaciation and uplift have contributed to Yellowstone’s unique features, it is what lies beneath the park’s surficial beauty that makes it appealing to bioprospectors

(see Figure 13). The 2.2 million acres comprising Yellowstone National Park are situated above a giant hotspot, or mantle plume. A hotspot or mantle plume is a geological term for a thinning in the earth’s crust. At such places the crust is thinner than usual and magma wells up toward the surface from the earth’s interior. The Hawaiian Islands sit over a well-known hotspot. As the Pacific plate moves over this hotspot, magma spews out of the crust, creating islands (Smith & Siegel 2000; Yellowstone National Park 2012).

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Figure 13: Yellowstone National Park as seen through visible light (left) and infrared (right). Bright areas in the infrared image indicate surface and sub-surface heat. Image: NASA’s Goddard Flight Center, 1999.

Yellowstone sits on a similar feature. In the present era, volcanism does not take place. But at one time in the Earth’s past it did. Over the past two million years, three massive volcanic eruptions have taken place in Yellowstone. The youngest of the three calderas erupted about 630,000 years ago. These eruptions dwarfed the eruption of Mt.

St. Helens in 1980. Geologists expect one or more of the calderas to erupt at some point in the future (Smith & Siegel 2000; Yellowstone National Park 2012).

This geologic activity beneath the placid exterior of Yellowstone provides a

desirable environment for thermophiles. Surface water percolates down where it contacts magma. The magma superheats the water. The heated water then moves upward, but it cannot escape to the surface. Much of it remains trapped in underground chambers at temperatures of over 204 ºC (400 ºF). However, at numerous places this superheated water moves through rock fissures to erupt at the surface; an event we call a geyser.

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Other superheated water makes its way to the surface in less spectacular manner, and accumulates in heated pools (Smith & Siegel 2000; Yellowstone National Park 2012).

Aside from geysers, Yellowstone harbors a number of different types of thermal features that interest microbial bioprospectors (see Appendix C). Fumaroles (also known as solfataras) are similar to geysers, but most of the water evaporates before reaching the surface. What escapes the underground vent is mostly steam or a mixture of gases. The escaping steam or gas can be so thick that it makes the ground look like it is smoking.

Then there are mud pots. Mud pots result from highly acidic water making its way to the surface. The acidic water dissolves the neighboring rock. Since there is not enough water to wash away the dissolved rock, a bubbling mass of silica and clay results. Superheated thermal pools occur in Yellowstone where the rock is not hard enough to resist the force of boiling water below, and the water bubbles out. These heated pools often occur in conjunction with thick layers of calcium carbonite (CaCO3). These superheated pools can be seen trickling over formations of travertine. Then there are the hundreds of other hot springs bubbling to the surface (Smith & Siegel 2000; Yellowstone National Park 2012).

Yellowstone’s natural wonders inspire awe in tourists and bioprospectors alike.

Bioprospecting in America’s Natural Parks: An Activity Reborn

Bioprospecting is not new to America’s national parks. It first started in YNP in

1898. The first bioprospector to obtain a permit was from the University of California.

W.A. Setchell collected specimens of thermophilic algae in Mammoth Hot Springs (Biel

2004; ten Kate, Touche, & Collis 1998; Varley & Scott 1998). In 2014, YNP granted 177 research permits for various purposes. Fifty-four percent of the research projects involved biological resources, including microbiology (Yellowstone National Park 2015).

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Two pieces of legislation facilitated bioprospecting in the national parks. The

Federal Transfer and Technology Act of 1986 encouraged federal laboratories and research facilities to enter into licensing arrangements with private contractors so that the government could benefit financially from intellectual transfers. On the heels of the

Yellowstone-Diversa CRADA, the National Parks Omnibus Management Act of 1998 allowed the Department of Interior (which oversees the NPS) to make benefit sharing arrangements with research institutions and private companies (Bryson & Kaczmarek

2000).

Chronic park funding deficits and the attention on the revenues that biological resources could bring, particularly in the wake of the Taq polymerase breakthrough, helped spur the drive to shape benefit sharing arrangements (Wood 2000). The NPS had its eyes on what was transpiring south of the border. The agreement with Diversa was designed to mimic the principles of the Convention on Biological Diversity—mainly, the conservation and protection of biodiversity, including sustainable use, and benefit sharing from the use of genetic or biological resources (ten Kate, Touche, & Collis 1998).

The National Park Service certainly has an argument for benefit sharing. Around the time of signing the CRADA deal, the NPS had a standing request of $8.7 billion to fund maintenance costs ranging from infrastructure to employee housing. Normally, national parks submit all revenues outside of entrance fees and recreation fees to the US

Treasury (Mattix 1999). Signing a profit sharing agreement under CRADA guarantees that the park in which bioprospecting occurs some of the revenue (Doremus 2004).

Under the CRADA, Diversa agreed to pay YNP $100,000 in five installments of

$20,000 over a five-year period (ten Kate, Touche, & Collis 1998). If the research

142 produced a commercial product, YNP would receive a royalty schedule of 0.5 to 10 percent. No actual number was made public (Doremus 1999).Yellowstone National Park received revenues from the actual signing of the deal. Any royalties were to be shared with the U.S. Treasury (Doremus 2004). Diversa also agreed to donate to YNP equipment

(DNA extraction kits) and staff training in PCR technology, estimated at $75,000 per year. Yellowstone National Park was to fund a Yellowstone Thermophile Conservation

Project program to encourage the research and conservation of Yellowstone’s microbial communities (ten Kate, Touche, & Collis 1998).

Under the arrangement allowed by the FTTA, Yellowstone was slated to pocket all revenues received through its CRADA deal with Diversa Corporation, as long as revenues did not exceed five percent of the park’s annual budget. That could have been about $1.4 million based on the approximately $28 million-dollar annual budget at the time. According to the CRADA, if revenues did not exceed the five percent limit, YNP was to retain 25 percent of the revenues, while the Federal Treasury would receive the remainder (Doremus 2004).

The contract between YNP and Diversa Corporation was nonexclusive (as are all benefit sharing agreements in the parks), so YNP was free to cut similar deals with other companies (which it has). Diversa Corporation did not gain (nor do other parties in other agreements) exclusive rights to any part of YNP (Doremus 2004). It is important to note that biological resources or specimens are not being sold. Only the intellectual knowledge and processes derived from such research (Yellowstone Association 2015).

Should another Taq polymerase novelty be extracted from its thermal pools, YNP could make hundreds of thousands in royalties. Some estimate that if YNP had been

143 collecting royalties from the revenues of the Taq polymerase, it could be collecting upwards of $400,000 to $3 million per year (Doremus 2004). Certainly something to get excited about, but YNP operates with a continuing budget deficit. The 2014 fiscal year budget for YNP was $69.7 million (Yellowstone National Park 2015). Even with substantial revenue sharing flows, YNP is unlikely to ever be self-supportive. Given

Yellowstone’s monopoly on thermal features and the diversity therein, it is doubtful other parks would come close to such a large royalty schedule. Like benefit sharing in the developing world, benefit sharing in the developed world is fraught with uncertainties.

The National Park Service was created in 1916 with a mandate from Congress to

“ . . . conserve the scenery and the natural and historic objects and the wild life therein and to provide for the enjoyment of the same in such manner and by such means as will leave them unimpaired for the enjoyment of future generations” (Yellowstone National

Park 2015, 23).

The benefit sharing agreement with Diversa was seen as throwing open the door to commercial exploitation of the parks that would erode this mandate and change fundamentally the future management of national parks. Others argued it seemed odd to allow commercial operators to remove biological material from national parks while the public faces fines for doing the same. The Edmonds Institute challenged the CRADA in court (Wood 2000). Like the CBD, what at first seemed like a win-win for everyone was turning into a quagmire.

Much of the debate centered on YNP qualifying as a “laboratory.” The FTTA allows federal laboratories to enter into CRADAs with private entities. Opponents to the

CRADA argued that the FTTA’s definition of laboratory “ . . . as a facility or group of

144 facilities owned, leased, or otherwise used by a Federal agency . . . ” did not fit the purpose of the CRADA. Based on legal precedence established by the Supreme Court, the NPS responded it had great discretion in interpreting “laboratory.” Ultimately, the courts agreed (Wood 2000).

The YNP-Diversa CRADA was finally greenlighted by the courts provided that the NPS complete an Environmental Impact Statement (EIS). That was finalized in

December 2009 (Lucchi 2013). By then, Diversa Corporation had lost interest in

Yellowstone, citing frustration with the legal road blocks (Dalton 2004).

Bioprospecting in Yellowstone and other national parks continues to move forward. Although requests for research permits continue, the majority of researchers are from public institutions, such as Montana State University’s Thermal Biology Institute.

Lucigen Corporation is a rare commercial bioprospector (Frank 2010). The company is currently awaiting approval from the Food and Drug Administration for an Ebola field test. The test is based on an enzyme extracted from a thermophile discovered in a thermal pool in YNP. The enzyme is not designed as an antivirus to Ebola, but rather to immediately identify those infected with Ebola. Instead of sending samples to a distant lab, medical personnel would be able to perform a test in the field and obtain the results within an hour (Newman 2014).

Bioprospecting has also moved underground. Carlsbad Caverns and Mammoth

Caves are of interest to extremophile bioprospectors (Doremus 1999). So is Lassen

Volcanic National Park in California. It is part of an extensive underground geologic complex that extends up to Crater Lake National Park in Oregon (Brown & Wolfe 2006).

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Outside of America, places like Russia’s Kamchatka Peninsula (Hoffman 2001) and Lake

Baikal (Hunter-Cevera, Repin, & Torok 2000) are targets of microbial bioprospecting.

Yellowstone and Beyond: Bioprospecting or Bio-destruction?

As America moves forward with bioprospecting in its national parks, it is worthwhile to examine differences and similarities between America’s national parks and those in the developing world. A major difference is the absence of indigenous peoples in

U.S. national parks. Bioprospecting in developing countries is often problematic because it introduces an additional set of actors into the narrative—that of indigenous peoples. As we have already seen, indigenous peoples may reside on lands of great biodiversity where bioprospecting is the most productive. Questions arise as to with whom should benefit sharing revenues be divided: the host government or the indigenous peoples. And with what groups of indigenous peoples should revenues be distributed.

America does not have this problem for the most part, as indigenous peoples were removed forcibly (if not brutally) from areas later designated as national parks. This process of removing indigenous peoples from national parks was, ironically, later known as the Yellowstone Model (Schelhas 2001).

Other questions have been raised. Doremus (1999) pointed out that there is a difference between the long history of the NPS issuing a permit for scientific research or bioprospecting, and a permit for commercial bioprospecting. For the past 20 years, since the CRADA was signed with Diversa, little public attention has been paid to microbial bioprospecting in America’s national parks. Seeing a steam shovel removing earth in

Yellowstone’s mud pots would certainly awaken the public to bioprospecting activities.

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The innocuous activity of removing vials of water from thermal pools is unlikely to ignite outrage.

Bioprospecting in America’s National Parks: Concerns and Issues

That raises the question—exactly what does this bioprospecting in national parks entail? Will pumps be sucking dry thermal pools? How much biological material will be removed? Opponents construct an image of wholesale plunder, while supporters paint a picture of minimal damage. It is probably somewhere in between.

Most research samples involve small amounts of organic matter, such as a few test tubes of water. Lucigen CEO David Mead may have over-simplified the question when he argued Lucigen and other researchers were only removing material that would otherwise be washed out to the rivers and the sea. In Lucigen’s case for the Ebola enzyme test, the amount of water removed from thermal pools was about two liters, containing a minute matter of DNA material (Frank 2010). Multiply that by dozens, hundreds, or conceivably thousands of researchers doing the same, and the possibility of damage to the environment or thermophilic habitats at some level, is within the realm of possibility. But

Lucigen’s Mead counters with a valid point that hordes of tourists can cause extensive damage to the national parks through weight of numbers (Frank 2010).

Another concern is that a thermophile with commercial applications may actually be discovered. Synthesizing the enzyme could require huge amounts of natural source material; enough so that it could potentially threaten the survival of the thermophile in its natural habitat (Doremus 2004). Given the technology now available to replicate DNA, this is probably unlikely. But again, no one knows exactly what microbe could be found.

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Perhaps one that cannot be replicated and would require huge amounts of natural biological material to be extracted.

The secrecy regarding the specific economics of the YNP-Diversa CRADA has caused concern among some groups. The amount of royalties to be returned (should a product return any revenues) remains unknown. The NPS was criticized for not divulging more details of the deal. Critics cited two major concerns with secret negotiations. First, a favorable royalty return may not have been secured. Second, secrecy denies the true owners of the park, the American public, from judging the merit of such deals (Doremus

2004). The NPS defended the secrecy, noting that many commercial biotechnology companies are hesitant to provide confidential information that competitors may find useful. Lawsuits were filed by the Edmonds Institute under the Freedom of Information

Act to obtain the details of the deal. The lawsuit was settled in 1998, but royalty schedules were still not made public (ten Kate, Touche, & Collis 1998).

An NGO called the World Foundation for Environment and Development

(WFED) advised the NPS in negotiating the CRADA with Diversa (Varley & Scott

1998). Reportedly, WFED received $28,000 for helping negotiate the deal (ten Kate,

Touche, & Collis 1998). How the WFED came to get the contract and how much it received is unclear. Given the national parks are owned by the American people there is reason for negotiations to be as transparent as possible to avoid improprieties. NGOs like the Public Employees for Environmental Protection (PEER) have accused Yellowstone

National Park of the illegal use of revenues derived from communication cell towers placed inside the park (Public Employees for Environmental Responsibility 2004).

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Failure of the NPS to regulate revenues generated from bioprospecting could generate similar complaints, and turn the public against benefit sharing arrangements.

There is also the concern with National Park Service personnel touting the financial gains to be made. It raises the question that the NPS may be making the mistake of the CBD and promising the public economic rewards that may never materialize.

Legal scholar Holly Doremus (2004) has raised similar issues. Should researchers hit the extremophile lottery and create a commercially rewarding pharmaceutical or other product, this could act to put more pressure on national parks to be self-sufficient.

Royalty distributions can lead to problems. If a substantial amount of money is returned to the Treasury, this could be too tempting for politicians to resist, and thus find ways to channel the money into other programs. These possibilities exist, but given the many watchdog organizations in place, it seems unlikely there will be extensive corruption or damage to the national parks through microbial bioprospecting agreements.

Chapter Summary

Despite some 24 thermophiles being isolated in Yellowstone’s thermal pools over the past few decades, Thermus aquaticus is the only microbe that has garnered public attention. More pharmaceutical and medical applications could be on the horizon.

Sulfolobus, a thermophile isolated from Yellowstone’s thermal pools, may provide a better insight on how the AIDS virus attacks proteins in the body. Then there is

Mammoth Hot Springs, which may hold fossilized microbes 33,000 years old embedded in its travertine rock (Yellowstone Association 2015).

Still, almost 20 years on since the announcement of the YNP-Diversa CRADA, no Taq polymerase-like microbe has emerged from YNP’s thermal pools. Although

149 microbial bioprospecting and benefit sharing is going forward in America’s national parks, hot gold may prove just as elusive as green gold.

Although laws are in place in the United States to regulate microbial bioprospecting for thermophiles and extremophiles, there are places in the world where such bioprospecting is moving forward without regulation or governing guidelines. In these places microbial bioprospecting is well ahead of any regulations and possibly could ignite geopolitical confrontations. These places exist in Antarctica and the maritime vastness of the planet’s oceans.

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CHAPTER 12

BIOPROSPECTING IN THE BLUE ABYSS

Finding Pharmaceutical Treasures at the Bottom of the Sea

Chapter Introduction

It is a commonly repeated mantra that humans know more about the moon than life in Earth’s oceans. About 70 percent of the planet’s surface is covered by a marine environment (Dionisi, Lozada, & Olivera 2011). That might seem prohibitive to bioprospecting, but it opens a vast new door as far as microbial organisms are concerned.

This chapter examines the potential of microbial bioprospecting in the pelagic vastness of the oceans, and particularly at the bottom of the sea floor.

Bioprospecting for Blue Gold

Marine microbes comprise 95 percent of all biomass in the ocean. Withdraw a liter of water from the ocean and you have captured millions of microbes (Abida,

Ruchaud, Rios, Humeau, Probert et al. 2013). Within that liter of water could be a biodiversity that dwarfs anything found among terrestrial life forms. That diversity is a result of billions of years adapting to environments where light, pressure, and temperature vary tremendously. Bright sunlight at the surface is replaced by growing blackness as one descends in depth. Atmospheric surface pressure increases to the equivalent of over 1,000 atmospheres (110 MPa) in the deep ocean. Ocean water temperature can range upwards

151 of 350 ºC (662 ºF) around deep sea vents to -35 ºC (-31 ºF) in Antarctic ice (Dionisi,

Lozada, & Olivera 2011).

Until the past few decades it was thought that the high pressures, cold temperatures, and lack of nutrients of the deep ocean offered no suitable habitat for life.

Now that technology has allowed humans to explore the ocean depths once off limits, researchers believe that marine microbial life could equal the biodiversity of rainforests

(Jobstvogt, Hanley, Hynes, Kenter, & Witte 2014). Microbes that inhabit these environments have developed resilient survival mechanisms. Largely for these reasons, many scientists are hopeful marine organisms can eventually provide numerous therapeutic drugs (Bhatia & Chugh 2015).

Marine bioprospecting is less than a hundred years old. It got off to a less than promising start. The academic community is divided on who should be regarded as the father of marine bioprospecting. Two people stand out as primary candidates. In 1945,

Guiseppe Brotzu was puzzled as to why Sardinia’s residents contracted fewer cases of typhoid than elsewhere, despite their habit of swimming in the sea near where the city’s sewage effluence was released. Brotzu took samples of sea water in the area and isolated

Cephalosporium acremonium. The microbe appeared to be resistant to Salmonella typhi.

Brotzu could not interest any Italian pharmaceutical companies in his discovery. His research later came to the attention of Eli Lilly. In 1964, Eli Lilly released an antibiotic under the brand name Cefalotin. Werner Bergmann and his fellow researchers also spearheaded the discovery of a pharmaceutical drug from marine environments. They isolated a nucleotide from Cryptotethia crypta, a sponge found in the Caribbean. It was refined into an effective anti-tumor drug (Armstrong, Falk-Petersen, & Kaspersen 2013).

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Microbes found in the ocean are attracting attention for three reasons. The first is a higher biodiversity than what can be found on land (de la Calle 2009). Some estimate that the ocean contains 3.67x1030 microorganisms with the vast majority of those microbes having never been identified (Synnes 2007). Since marine bioprospecting in earnest has been taking place for only two decades, there is more virgin territory to mine than on land (de la Calle 2009).

The lack of light, extreme temperatures (both hot and cold), crushing pressures, salinity, acidity, and varying degrees of nutrients, all acting in combination, have forced marine microorganisms to adapt to a variety of extreme environments (Synnes 2007). In turn, they have evolved unique metabolites often not found in terrestrial microbes

(Dionisi, Lozada, & Olivera 2011).

Thirdly, marine organisms have shown a history for higher positive hit rates than terrestrial organisms. A study by the National Cancer Institute found that terrestrial organisms returned a positive hit rate of 0.01 percent while marine organisms returned a

0.1 percent positive hit rate. Already 18,000 marine metabolites have shown uses in medicinal applications. (de la Calle 2009). Others report that the rate for positive hits from marine organisms is 500 times higher than that of terrestrial microbes (Dionisi,

Lozada, & Olivera 2011).

However, marine bioprospecting, while offering great possibilities, is fraught with great hurdles. Marine bioprospecting is extremely expensive. For that reason it is often a collaboration between the public and private sectors. Research vessels and submersible equipment with sophisticated technology are required when journeying to deep sea floor vents (Armstrong, Falk-Petersen, & Kaspersen 2013). So far, most of the collection of

153 marine microorganisms has been done at the upper levels of the ocean near the surface, where it can be done more quickly and more cost effectively (Hunt & Vincent 2006).

Until the past few decades, the oceans were not seen as holding much potential for microbial investigation. Researchers believed that the marine environment contained few microbes due to its salinity. Even after this was found to be otherwise, marine microbes were considered too challenging to isolate in a laboratory until recent advancements in biotechnology (Dionisi, Lozada, & Olivera 2011).

A third development made bioprospectors reconsider the microbial potential of the deep marine environment. In 1977, an accidental discovery suddenly focused attention on the sea floor thousands of feet beneath the surface. Researchers from the

Woods Hole Oceanographic Institution (WHOI), using the deep submersible vehicle

Alvin, found an unexpected and astounding geomorphic oddity on the sea floor along the

Galapagos Rift. Thousands of feet beneath the surface super-heated water was being spewed from the crust of the earth. These geologic features became known as marine hydrothermal vents (MHV), or more colloquially, as black smokers (Woods Hole

Oceanographic Institution 2015). See Figures 14, 15, and 16.

Most places where MHVs are found are locations where crustal plates are spreading, creating new sea floor. In other places, oceanic plates are subducting, creating deep trenches on the ocean floor. The majority of these undersea oceanic ridges are connected. If the oceans were drained, it would resemble a 40,000-mile suture stretching helter-skelter around the Earth (Allen 2001; Leary 2004; Rameriz-Llodra, Shank, &

German 2007). See Appendix D for additional images of deep sea vents.

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Figure 14, left. Researchers from WHOI photographed the first hydrothermal vent known as a “black smoker” ever seen by humans. From NOAA Ocean Explorer, photograph courtesy WHOI, 1979. Figure 15, top right. A white smoker. Figure 16, below right. A black smoker. Photographs by German/WHOI, 2013.

Marine hydrothermal vents function much like thermal vents found in

Yellowstone National Park. Sea water seeps into fractures in the oceanic crust where it is super-heated by magma. The super-heated water then erupts from vents on the ocean floor in a plume of black fluid that looks like smoke. Temperatures of these thermal vents can be a shocking 400 ºC (752 ºF)—so hot that the water can melt lead. The water contains a mixture of metals and chemicals making it a hot broth for life (Allen 2001;

National Oceanic and Atmospheric Administration 2013; Rameriz-Llodra, Shank, &

German 2007; Synnes 2007; Woods Hole Oceanographic Institution 2015).

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Marine hydrothermal vents, or black smokers, get their name from the color of their “smoke” which is a mixture of superheated metals, chemicals, and water. The black smoke is actually a thick broth of fine sulfide particles that precipitate when the superheated water mixes with the extremely cold seawater at the ocean floor. The precipitated sulfide-iron chemicals solidify into polymetallic sulfide chimneys. White smokers exist.

These vents operate the same as black smokers but do not emit dissolved metals. White smokers disgorge a lighter-colored chemical broth containing barium, calcium, silicon, and other chemicals (Allen 2001; National Oceanic and Atmospheric Administration

2013; Rameriz-Llodra, Shank, & German 2007).

Sunlight-adapted organisms produce energy through photosynthesis. But thousands of feet beneath the surface in total blackness, whole communities of organisms were found to be producing energy through chemosynthesis. That is, extracting chemicals in the water for energy. Thriving communities of tubeworms and other previously unknown organisms, both macroscopic and microbial, were discovered thriving in the vicinity of marine hydrothermal vents (Allen 2001; Leary 2004; Woods Hole

Oceanographic Institution 2015).

Since the discovery of hydrothermal vents in 1977, over 100 areas containing marine hydrothermal vents have been discovered with at least 550 new vent species identified. Two to three new species are added to that list each month (Rameriz-Llodra,

Shank, & German 2007). The deepest vents discovered are those in the Marianna Trench at 11,035 meters (~36,000 feet) beneath the surface (Leary, Vierros, Hamon, Arico, &

Monagle 2009).

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The diversity of locations with varying pressures, sunlight, and temperatures, creates a corresponding biodiversity around marine hydrothermal vents that has earned them the nickname “oases of the abyss” or “oceanic gardens of Eden” (Leary 2004, para.

7). Up to 90 percent of the 500 species identified living near MHVs were previously unknown because they are endemic to these strange environments (Leary 2004). The biodiversity of MHV microbes differs dramatically from one deep sea vent to the next because up to 75 percent of organisms that thrive around MHVs are endemic to just that one area (Leary 2004; Rameriz-Llodra, Shank, & German 2007).

The biodiversity of marine microbes is not limited to marine hydrothermal vents.

Cold seeps on the ocean floor also present bioprospecting opportunities. Cold seeps are often found on continental margins. In these peculiar places methane mixes with hydrogen-rich chemicals like hydrogen sulfide that seep from ocean sediments. The extreme pressure and near-freezing temperature creates an environment favored by microbial organisms that thrive in such radical conditions (Synnes 2007). Macrobial and microbial marine organisms together have been the source of over 15,000 new chemical compounds since the 1970s. Sixteen anti-tumor compounds being clinically tested were derived from marine microbial sources. When the number in pre-clinical trials is included it pushes the number closer to 30 (Xiong, Wang, Hao, & Wang 2013). Cephalosporin

(antibiotics), Cytarabine (leukemia treatment), and Vidarabine (treats eye infections), are well-known drugs derived from marine environments that are already on the market

(Haefner 2003). Ziconotide (pain relief); Trabectedin (aka Yondelis; liposarcoma or leiomyosarcoma treatment); Eribuline mesylate (cancer treatment); Omega-3 fatty acids; and Dolastatin (anti-cancer tumors) are more well-known FDA-approved drugs with

157 marine origins (Bhatia & Chugh 2015; Drugs.com 2015). Seven therapeutic drugs have been derived from marine environments as of 2012. One is in Phase 3, which is the final stage before being released to the market. Six are in Phase 2 clinical trials (Armstrong,

Falk-Petersen, & Kaspersen 2013). See Table 3.

Table 3. A Sampling of Current Pharmaceutical Products Derived from Marine Organisms

Originating Category Product Use Organism Status Revenues of Ecteinascidia Therapeutic Yondelis Anti-Cancer €20 million as turbinata (Ascidian) of 2012 Revenues of Neuropathic Conus Therapeutic Prialt $20 million as pain magus(mollusc) of 2012 Revenues of Halichondria okadai Therapeutic Halaven Anti-Cancer $200 million (Sponge) as of 2011 Salinispora tropica Phase 1 Therapeutic Salinisporamide Anti-Cancer (Bacterium) clinical trials Aspergillus sp. Phase 1 Therapeutic Plinabulin Anti-Cancer (Fungus) clinical trials UV Mycosporine-like Beginning Sunscreen absorbing Zooxanthellae (coral) amino acids trials potentials Pseudopterogorgia anti- In commercial Cosmetic Pseudopterosins elisabethae (soft inflammatory use coral) Thermus anti-free In commercial Cosmetic Venuceane thermophilus radicals use (Bacterium) Omega-3 fatty Crypthecodinium In commercial Nutrition acids cohnii (microalga) use Dunaliella salina In commercial Nutrition Cartenoids anti-oxidant (microalga) use Note: Adapted from Global Ocean Commission, 2013.

Pharmaceuticals are not the only possible use. Useful microbes have already been plucked from the ocean depths for industrial applications. One microbe inhabiting a black smoker is the source of an enzyme used in producing corn ethanol. This was due to the

158 discovery of the mother microbe flourishing at a pH of 4.5 in a deep ocean vent—an acidic level optimum in ethanol production. Another thermophile has yielded an enzyme useful in oil and gas extraction (Sheridan 2005).

Plankton, a microbial organism which has already produced a number of secondary metabolites that have generated over $200 million in revenues for cosmetic products, could become equally useful in agriculture. Plankton microorganisms live in low-nitrogen environments. Splicing their DNA with plants used in terrestrial agriculture could produce plants that require less nitrogen (Abida et al. 2013). A conceivable benefit would be the reduced need for nitrogen fertilizers, thereby reducing nitrogen runoff into waterways.

Marine Bioprospecting: The Cost Barrier

For a long time pharmaceutical companies were hesitant to enter into marine bioprospecting. The deep oceans were considered something of a biological desert

(Jobstvogt, Hanley, Hynes, Kenter, & Witte 2014). The largely unknown marine environment was considered too risky for pharmaceutical bioprospecting. Pharmaceutical companies decided to stay the course with developing drugs from terrestrial sources.

Thus, pharmaceutical companies were content to invest money into the terrestrial bioprospecting of plants which reached an estimated $9 billion annually in the 1980s. In the late 1980s and early 1990s, the technology of combinatory chemistry and high- throughput mass screening inspired great hopes for advances in synthetic compounds.

Natural products like plants, and even more expensive to obtain marine products, looked even less appealing. The drive to develop a blockbuster pharmaceutical helped slow the entry into marine bioprospecting and drug development. Driven by stockholder pressures,

159 pharmaceutical companies went through a period of corporate mergers (Glaser & Mayer

2009).

As private pharmaceutical companies seemed to be turning away from marine bioprospecting, public researchers were becoming more interested. The National Cancer

Institute announced that marine products returned a better hit rate in bioassays than that of terrestrial biological products. Funding for academic research, particularly for researching anti-cancer drugs from marine products, increased. Although private companies maintained some degree of research into drugs from marine biological products, drug development remained largely in the laboratories of academia. Even in recent years it remains so (Glaser & Mayer 2009).

This is an important departure from previous drug development that utilized terrestrial sources like plants. When it comes to marine bioprospecting, Big Pharma appears to be content to let others take the initiative—and risk—of drug development from marine sources. There are private pharmaceutical companies that do maintain an active presence in marine bioprospecting. However, drug development from marine sources, be it microbial or otherwise, remains largely in the hands of academic researchers, government research laboratories, and private venture capital biotechnology firms. Large pharmaceutical firms typically step in with infusements of cash and sign in- licensing deals with smaller companies and research institutes once initial screening and devleopment has shown promise (Glaser & Mayer 2009).

The reluctance of pharmaceutical companies to shoulder all of the research is understandable when one considers the enormous capital outlay required to perform marine microbial bioprospecting. Costs of a deep sea venture can reach $30,000 a day or

160 up to $1 million per month. Then it can take $1 billion in research and development over

15 years to bring a marine pharmaceutical to market (Global Ocean Commission, 2013;

Ruth 2006). Leary, Vierros, Hamon, Arico, & Monagle (2009) did not find any substantial documentation of research missions to the deep sea for the purpose of collecting genetic resources by a private company.

This hesitation to initiate research on the part of large pharmaceutical companies is why the National Cooperative Drug Discovery Group (NCDDG) was formed by the

NCI as a consortium between drug companies like Bristol Myers Squibb and Novartis, plus several university campuses across the country. For the most part, pharmaceutical companies have been content with this arrangement (Glaser & Mayer 2009). Yet interest in marine bioprospecting is there. Global sales from marine microorganism-derived products is expected to hit US $4.9 billion by 2018 (Global Industry Analysts, Inc. 2013, cited in Wynberg 2015).

Despite enormous capital outlay and other challenges, once in the lab the process of bioprospecting for marine microbes is similar to its terrestrial counterpart. So is the extreme cost, the time of development, and the miniscule odds of success (Glaser &

Mayer 2009; Hunt & Vincent 2006). However, interest is kept alive in marine bioprospecting by successes like Yondelis, an anti-cancer drug extracted from a

Caribbean Sea slug, and Prialt, a pain medication synthetically derived from the venom of a marine cone shell (Leary, Vierros, Hamon, Arico, & Monagle 2009).

Although sizeable revenues from commercial uses of marine microbes and marine biotechnology remain elusive, a single blockbuster success will open the door to further interest and investment, much the way Farnsworth’s list helped to stimulate plant

161 bioprospecting several decades ago. This will create problems that are more politically tense than bioprospecting between developed and developing countries. That is because the vastness of the ocean is not a developed-developing world issue. Rather, potential conflicts will be between wealthy, developed, and militarily strong countries.

Bioprospecting on the High Seas and the Ambiguities of UNCLOS

In terrestrial bioprospecting it can be difficult to draw a line between where one resource (a plant) begins or ends. So it is with microbial bioprospecting in the deep oceans (known as the High Seas, or the Area). Although maritime countries are each granted an exclusive economic zone (EEZ) that extends 200 nautical miles from their shore, this does little to deal with legalities governing deep sea vents thousands of miles offshore, or mobile organisms that may swim or drift into and out of the EEZ.

The microorganism plankton shows promise for both pharmaceutical and industrial applications. Plankton is a derivative of the Greek word planktos, or drifter.

Because marine plankton drifts with the currents, a microbe with commercial applications discovered in international waters could in fact be found drifting within a certain country’s territorial waters. This could create international legal disputes over ownership

(Abida et al. 2013). Would a country that engineered the plankton microbe into a commercial product receive sole revenues, or would that country have to share it with all maritime countries? If oceans are regarded as the common property of all countries, would every country participate in revenue sharing? And how would this revenue sharing arrangement be distributed? By country population size, length of coastline, or otherwise? The legal implications are numerous. Unfortunately, international maritime law provides little clarification.

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The United Nations Convention on Law of the Seas (UNCLOS) was adopted on

November 15, 1994, after a multi-decade’s quest to find the means of governing the largely here-to-fore unregulated vastness of the ocean (Allen 2001; United Nations 2013).

The first attempt to establish guidelines in the open ocean was in 1945 when President

Truman issued the Proclamation on the Continental Shelf. This was followed by the 1958

Convention on the Continental Shelf. The 1982 Law of the Sea Convention, which ultimately gave birth to the UNCLOS, supplanted the 1958 Geneva Conventions Law of the Sea (Allen 2001).

Under UNCLOS, each maritime country is granted a 12 nautical mile territorial zone that gives a state complete sovereignty over access to waters and resources within those waters, plus the sea floor. A 200 nautical mile EEZ allows a country to regulate marine resources and activities within that area. The Continental Shelf Zone allows countries the right to explore and exploit sea bed resources within 200 nautical miles (see

Figure 17). The ocean vastness, known as The Area (a.k.a. the High Seas) does not allow for national jurisdiction or sovereign claims (Leary 2004). Where exactly the Area begins is unclear because states may claim their continental shelf extends beyond the EEZ

(Arico & Salpin 2005). Therefore, deciding where to draw the line on a marine resource can be as difficult as determining what group can claim TEK rights to a plant.

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Figure 17. Exclusive Economic Zones (hashed areas) as demarcated by UNCLOS. The Area/High Seas is unregulated, creating an opportunity for tensions between developed countries over bioprospecting resources. EEZ shapefile from Marine Regions, Flanders Marine Institute, 2014; Base map from ESRI, 2014.

How UNCLOS applies to contemporary marine bioprospecting is unclear. Like the CBD, the document does not use the term bioprospecting (Arico & Salpin 2005). The

Nagoya Protocol supposedly streamlines access to marine genetic resources, but it is too early to know if it accomplishes that goal (Burton & Evans-Illidge 2014). Nor does

UNCLOS discuss genetic resources (Arico & Salpin 2005). This is because when

UNCLOS was conceived it focused on mineral and hydrocarbon extraction, not regulating the extraction, uses, and patenting of biological or genetic resources (Sheridan

2005). It has not been resolved if the 200 nautical mile EEZ gives maritime or coastal

164 countries the legal right to regulate bioprospecting of marine resources, including hydrothermal vents, within the EEZ (see Figure 18).

Figure 18. Boundaries of maritime sovereignty as demarcated by UNCLOS. From the Canadian Department of Fisheries and Oceans 2015, in Wynberg, 2015.

More uncertainty rests on how “sedentary species” is interpreted as defined in

Article 77 of UNCLOS, according to Leary (2004). A species may be sedentary at one stage in its life and not in another. Nor is it clear on how to handle disputes that may arise if the same species of microbe is discovered at a hydrothermal vent in international waters, and also within a country’s EEZ or territorial zone.

The International Seabed Authority (ISA) as designated in section 157 in part IX of UNCLOS is the governing body that oversees resource exploration and extraction in the Area. Yet “resources” are left undefined. The previous interpretation of resource was generally considered to govern mineral, oil, or natural gas extraction. It has not been determined as to how various sections of the document govern bioprospecting. The

165 question is further complicated by the Marine Scientific Research clause found in

UNCLOS. The High Seas Freedom provisions in Article 87 of UNCLOS allows states to conduct unfettered scientific research into resources in the Area (international waters).

Bioprospecting is not specifically mentioned (Leary 2004).

Other sections of UNCLOS lead to equally confusing interpretations. Article 136 provides that all resources within the Area be considered a common heritage of mankind

(CHM). Article 137 strictly prohibits national sovereignty over such resources. The problem is that UNCLOS does not adequately define what is meant by CMH (Allen

2001; Leary 2004). If a microbe is a common heritage of mankind under Article 136, does Article 137 then prohibit any one nation from extracting that microbe for pharmaceutical or industrial uses? Lack of clarity leads to more questions. Is a microbe a resource in its raw form, or a resource after it has been altered in the laboratory? The present reading of UNCLOS creates many dilemmas.

There is also uncertainty on how borders encompassing the EEZ or the

Continental Shelf should be drawn in regards to marine bioprospecting. If a nation delineates its EEZ boundary as a straight 200 nautical mile line, it could potentially push its sovereign authority into areas not recognized by other countries. Nor is it clear how to regulate MHVs that are split by a nation’s EEZ or Continental Shelf boundary. If a MHV field is split by such a boundary, does ownership of a MHV microbe belong to a single nation or is it up for grabs?

Other uncertainties further complicate the issue of marine bioprospecting. It is difficult to determine what constitutes marine scientific research, and what constitutes commercial bioprospecting (Arico & Salpin 2005; Ruth 2006). Since research of

166 hydrothermal vents for microbial bioprospecting can also be a precursor to exploration for mineral deposits, it is unclear as to how to draw lines between marine scientific research (MSR) of deep sea floor hydrothermal vents, and exploration for minerals, gas, or oil (Leary 2004; United Nations 1994). Duplicitous countries could argue they are performing research when in fact they are bioprospecting, thus avoiding any guidelines that might regulate such activities.

These questions will arise given what is at stake. Potential economic revenues from one MHV field in the Eastern Pacific were pegged at US $3 billion a year (Allen

2001). This much possible revenue, combined with the vagueness of UNCLOS, could lead to countries conducting activities of any nature, as long as it is under the banner of bioprospecting. Such activities could trigger confrontations between nations. Since the majority of the Area lies outside of national boundaries, and because of such is governed

(or not governed) by UNCLOS, addressing bioprospecting constitutes a pressing urgency.

Bioprospecting on the High Seas: No Guidance from the CBD

The Convention on Biological Diversity offers little assistance or clarification for bioprospecting in international waters. This is because the CBD is a framework for how bioprospecting activities should proceed, not how they must proceed. Each state has sovereign authority over how it implements the CBD. Thus, the CBD allows each country to implement the guidelines of the CBD in its own territorial waters. The CBD does not apply to the High Seas (the Area) in international waters. Therefore, neither

UNCLOS nor the CBD offer regulations or guidelines for bioprospecting at deep sea vents (Leary 2004).

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The CBD and UNCLOS differ in other ways. The CBD is generally recognized to emphasize conserving overall genetic resource biodiversity, while UNCLOS is concerned with conserving individual species diversity. The CBD urges ecosystem conservation while UNCLOS does not. When conducting research in a sovereign nation’s maritime waters, the CBD requires prior informed consent (among contracting parties) while

UNCLOS does not. The research or scientist-friendly leanings of UNCLOS would therefore conflict with bioprospecting benefit-sharing arrangements of the CBD. Since the CBD applies to a country’s EEZ if such is applicable, the CBD would probably trump

UNCLOS. However, for bioprospecting conducted in the Area it is accepted UNCLOS

(which provides no regulation for bioprospecting) would be the guiding document (Allen

2001; Leary 2004; United Nations Oceans & LOS 2015). Such inability for the CBD to regulate the exploitation of biological diversity in the deep sea is ironic, because genetic resources are the one deep sea resource that are the most readily available and potentially lucrative (Leary 2004).

The stakes for real conflict in the deep ocean are much greater than mere allegations of biopiracy. In previous bioprospecting disputes a lopsided balance of power existed between North and South, or developed and developing countries. Bioprospecting conflicts in the Area will be between militarized countries, such as Russia and the United

States, or Japan and China.

Discussions on marine bioprospecting threaten to alienate many poor, developing countries the same way terrestrial bioprospecting has done. Marine bioprospecting is an enormously costly endeavor and this could leave many developing countries that control long shorelines (i.e., Brazil) once again feeling they are on the losing end. These

168 countries may not be able to access potential commercially profitable microbes within what they consider their legal domain due to cost barriers.

To counter this, countries like Brazil are seeking to declare biological resources in international waters as a common heritage of mankind. This could force wealthier countries into benefit sharing agreements. The United States (which recognizes UNCLOS as codified international law, but has not signed) and Japan are reluctant to embrace such interpretations (Sheridan 2005). During negotiations for the Nagoya Protocol, it was decided to limit the geographical scope of the document to ensure it did not regulate bioprospecting in the Area (Buck & Hamilton 2011).

Given the wide latitude in interpreting UNCLOS, it is not surprising that charges of biopiracy and the theft of indigenous medicinal knowledge of marine organisms have already been made. Like allegations of biopiracy surrounding plants, allegations of biopiracy surrounding marine organisms are repeated with little scrutiny. Bhatia and

Chugh (2015) cite an allegation made by Someshwar Singh on the website Third World

Network. This website claims, “The plunder and patenting of marine life also has not abated (Singh 2000, para. 13).” The website (accessed January 2016) cites a report of a compound derived from the Caribbean sea-whip by University of California researchers that is useful as the anti-inflammatory agent pseudopterosin in skin cream. The website document claims the compound was “discovered” without researchers disclosing the country of origin (Singh 2000, para. 16). However, the soft coral in question is found in many Caribbean locations at depths of 31 meters (~100 feet) or more, off of countries such as the US Florida Keys, Nicaragua, Honduras, Colombia, Bahamas, Belize, and

Mexico (Huggan 2013; Onumah 2013). Establishing TEK ownership of marine microbes

169 will likely be more difficult than that of terrestrial plants. Allegations of biopiracy may follow bioprospecting to the bottom of the sea.

Yet marine bioprospecting may provide an opportunity for some developing countries to reset the clock, so to speak. The southern African coast is known to possess high levels of endemism of marine organisms (Wynberg 2015). By joining consortiums from wealthier countries, these poorer nations with a high offshore biodiversity may be able to profit from marine biological resources.

Chapter Summary

Bioprospecting for marine microbes has already shown that it will be different from terrestrial bioprospecting. Unlike an ethnobotanist or researcher setting out into the rainforest with minimal financial backing, bioprospecting in marine ecosystems, specifically deep marine areas, is enormously expensive. The great expense and cutting- edge technology required will certainly limit this new era of microbial bioprospecting to a select group of wealthy countries.

The issues raised by marine microbial bioprospecting may be more complex and more contentious than those created by bioprospecting for terrestrial biological products.

Differentiating who owns what and discerning who is doing what thousands of feet beneath the surface may create political or military confrontations. The complexities of marine microbial bioprospecting will probably be surpassed by only the third geography of bioprospecting’s new era—Antarctica. This is addressed in the next chapter.

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CHAPTER 13

BIOPROSPECTING FOR COLD GOLD

Bioprospecting in Antarctica

Chapter Introduction

It is doubtful that the explorers Roald Amundsen or Robert Scott would have guessed that the frigid environs of the Antarctic would become a hot spot for tourism. But in a little over 100 years since the race to the South Pole ended, the continent of

Antarctica has become a hub of research and recreation. Just a few years ago Antarctica was logging 33,000 tourists per year, with upwards of 100 permanent research facilities

(Hughes, Pertierra, & Walton 2013).

Hidden at the bottom of the Earth, many people do not realize the massive size of the Antarctic continent. Antarctica is equivalent to the United States and Mexico in size

(Zell 2014), or twice the size of Australia (Herber 2012). See Figure 19. Massive ice sheets averaging one mile in thickness cover large portions of the continent (Zell 2014).

Antarctica contains 90 percent of the world’s ice, with 70 percent of all freshwater locked in its massive ice sheets (Herber 2012; Zell 2014). Roughly only 0.34 percent of the landmass is ice free (Hughes, Pertierra, & Walton 2013). Antarctica might appear to be little more than a frozen wonderland to tourists, but to bioprospectors it offers opportunities to discover new microbes. Antarctica is considered an exotic environment.

Exotic locales generally mean unique biodiversity (Bowman 2001).

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Bioprospecting for Microbes at the Bottom of the World

Antarctica’s microbes are highly appealing to bioprospectors for two reasons.

With little bioprospecting having been conducted on the continent, Antarctica is largely virgin territory, and so are its microbes (Bowman 2001; Lohan & Johnston 2005). Polar extremophiles survive not only in unbearably low temperatures by human standards, but in frigid temperatures combined with the high salinity of ocean waters, or few nutrients, or limited sunlight. Microbes that survive in such extreme environments are thought to have evolved equally extreme enzymes (de Pascale, De Santi, Fu, & Landfald 2012).

Figure 19. A comparison of the landmass of Antarctica and the United States. Antarctica is shown using the Landsat Image Mosaic of Antarctica (LIMA). From Zell, 2014.

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Seawater, ice sheets, frozen lakes, soils, and fumaroles on and around Antarctica are home to cold-enduring microbial organisms whose enzymes may hold uses ranging from bioremediation, resistance to freezing, to other biochemical processes. Ice cores taken from ice sheets have yielded microbes termed ultra-microbacteria that date more than 100,000 years old. The Southern Ocean surrounding Antarctica is low in nutrients, but may yield microbes that lead to unique catalysts (substances that facilitate chemical reactions) and ligands (substances that bond to biomolecules). Antarctica’s continental shelf sediment is thought to hold an extremely high diversity of microbes (Bowman

2001).

Microbial bioprospectors are interested in the interior of Antarctica. The Ross

Desert has a temperature hovering near -20 ºC (-4 ºF) but its arid vastness holds cryptoendoliths. These are microbes that grow inside rocks. The McMurdo Dry Valleys and the region’s frozen lakes, situated in an ice-free zone of Antarctica where temperatures average -27.6 ºC (-17.6 ºF), are home to unique extremophilic microorganisms (Priscu & Christner 2004).

Antarctica’s lakes offer more bioprospecting opportunities. Deep Lake is found in

Eastern Antarctica’s Vestfold Hills (see Appendix E for map). Its chilly waters never climb above -10 ºC (14 ºF) and are ten times more saline than ocean water. Yet microbes thrive in its harsh environment. Lake Vostok lies buried 3.6 km (2.2 miles) beneath the icy Antarctic surface. It cannot be seen through the ice but its location is known through seismic imaging. Lake Vostok’s waters have been cut off from the current atmosphere for one million years. Molecular analysis combined with radioisotope tracers has discovered living microbes inside the lake (Bowman 2001).

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Antarctica is not all frozen. Fumaroles are located on Deception Island and other islands in the Shetland Archipelago (Hughes, Pertierra, & Walton 2013). Mt. Erebus, at over 12,000 feet, is an active volcano that has provided researchers with thermophilic microbes that survive in temperatures over 95 ºC (203 ºF) (Farrell & Duncan 2005).

Antarctica is not the only polar environment of interest. The polar biospheres comprise 14 percent of the Earth’s surface (Rampelotto 2014). Microbiota with a high diversity is expected to be found in extreme northern regions such as seasonal sea ice, the

Greenland Ice Sheet, and the tundra (de Pascale, De Santi, Fu, & Landfald 2012).

The evolution of polar microbial bioprospecting followed a story similar to the bioprospecting of plants for new compounds. The so-called genomic era that relied on high-throughput screening and DNA sequencing technology greatly expanded the field of polar microbiology (Rampelotto 2014).

Bioprospecting has been underway in Antarctica since the 1980s (Hemmings

2010). Until the 1950s, little was known about polar microbiota. As researchers penetrated the polar environments, they began to look for ways to elucidate the novel compounds produced by microbes living in these harsh environments (de Pascale, De

Santi, Fu, & Landfald 2012). Interest in applications for polar microbes began in the

1970s when an antifreeze extract was taken from a fish in waters around Antarctica

(Lohan & Johnston 2003). By the 1990s, automated gene sequencing and high- throughput screening greatly reduced costs, making it feasible to compile large genetic libraries unthinkable only a decade earlier (de Pascale, De Santi, Fu, & Landfald 2012).

Polar bioprospecting for industrial uses focuses on four core goals: obtaining enzymes that can be used in food production and processing (allowing crops and

174 packaged foods to resist cold); pollution control technologies (bioremediation); dietary supplements (cold-loving microbes manage their fatty acids extremely efficiently, and this could lead to dietary supplements that manipulate fatty acids in humans); and antifreeze technologies used in food processing (de Pascale, De Santi, Fu, & Landfald 2012).

The Antarctic Bioprospector, a United Nations database of bioprospecting operated by the United Nations University Institute of Advanced Studies, has logged over

185 commercial products derived from organisms located on Antarctica and in the

Southern Ocean (Leighton 2015). A species of Actinobacteria that includes Streptomyces, along with Nocardia and Micromonospora, are all members of a genus that has produced numerous bioactive compounds in the past (Lohan & Johnston 2005).

Bioprospecting in Antarctica is, like deep sea marine bioprospecting, an extremely expensive endeavor. Lohan and Johnston (2005) found that a handful of wealthy, developed countries that possessed cutting-edge technology held the majority of patents pertaining to Antarctic enzymes. Food processing applications held the largest number of patents with 27 percent. Enzymes with pharmaceutical applications held the second largest number of patents with 26 percent, with patents targeted at agriculture a close third with 23 percent. Japan held the largest number of annual patent applications at an average of seven per year, 1998 through 2003. Germany was second with half of that number.

Although the economic value from many of these patents is still unrealized (or undisclosed), it has not slowed international competition for microbes in Antarctica or the

Southern Ocean. A Russian company has patented a yeast found in Antarctica, while a

US company has patented an enzyme taken from krill and certain types of fish. Australia

175 is amassing a large stockpile of Antarctic microorganisms (Jabor-Green & Nicol 2003).

The Australian Collection of Antarctic Microorganisms possesses over 300 species. A private collection by the Cooperative Research Centre for the Antarctic and Southern

Ocean numbers over 7,400 specimens (Chaturvedi 2010; Jabor-Green & Nicol 2003).

Like its deep marine counterpart, bioprospecting in Antarctica is largely a joint public and private venture. A number of contracts between research institutes and private companies have already been concluded. One contract called for the Antarctic

Cooperative Research Center at the University of Tasmania, Australia, and AMRAD, a natural cosmetics pharmaceutical company, to jointly screen over 1,000 samples collected in the field. The goal is to discover useful pharmaceuticals (Lohan & Johnston

2003).

These contracts spread the risk and economic burden of bioprospecting in areas that require huge capital outlays. While spreading the expense and risks can encourage bioprospecting in Antarctica, these private-public consortia have the potential to merge scientific and commercial objectives (Johnston & Lohan 2005). As private companies look to recoup their investment, they will likely seek patents over any biological products sourced from Antarctica, along with claiming profits from products derived from those sources (Jabor-Green & Nicol 2003).

Bioprospecting in Antarctica and Geopolitical Tensions

Like its abyssal counterpart in the vast oceans, microbial bioprospecting is moving forward in Antarctica and the Southern Ocean with little regulation or oversight.

There is an absence of universal legal guidelines in place to ensure that microbial

176 bioprospecting in Antarctica will not repeat the same problems of bioprospecting in tropical climates.

At present, there is no framework regulating bioprospecting in Antarctica. At the time of the Antarctic Treaty’s adoption bioprospecting was not on the horizon and no guidelines were promulgated (Hemmings 2005). Little has been done since. Failure to act could leave Antarctica’s fate largely determined by market forces. This inaction is partly due to what Hemmings (2010) describes as a chronic resistance among nations when attempting to promulgate new guidelines for Antarctica. The previously held “Antarctic exceptionalism” is constantly being challenged by the demands of globalism (10). Since the Antarctic Treaty System regulates 10 percent of the Earth’s surface (Hemmings

2005), the lack of bioprospecting guidelines could lead to political disputes.

Much of the present geopolitics of Antarctica is a remnant of decaying geopolitical structures created decades ago during the Cold War. The Antarctic Treaty in

1959, and its subsequent Article IV, suspended resource exploration and extraction, along with banning all military activity and nuclear testing (Chaturvedi 2010; Leary 2008a;

Leary 2008b). The United States wanted to preclude any Soviet domination of Antarctica.

By 1949, the U.S. was pushing for Antarctica to be internationalized as part of its anti-

Soviet containment policy. The 1957-58 International Geophysical Year (IGY) and the push to view Antarctica more as a cold laboratory than a stage for the Cold War, provided much of the foundation for the 1959 Antarctic Treaty (Chaturvedi 2010).

It is important to note that under the ATS (Antarctic Treaty System), Antarctica is defined as all geography south of 60˚ latitude. This is different than the Arctic, which is generally accepted to encompass all geography north of 66.5˚ latitude (Rampelotto 2014).

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The Antarctic Treaty System was promulgated in 1959 and has been in effect since 1961. Since then, other conventions have been added to clarify and strengthen the

ATS. These include: the Convention for the Conservation of Antarctic Seals, effective

1972; Agreed Measures for the Conservation of Antarctic Fauna and Flora, effective

1982; the Convention for the Conservation of Antarctic Marine Living Resources, effective 1982; and the Protocol on Environmental Protection to the Antarctic Treaty, effective 1998. The Convention on the Regulation of Antarctic Mineral Resource

Activities never entered effect (Chaturvedi 2010; Hemmings 2005; Jabor-Green & Nicol

2003; Leary 2008a; Lohan & Johnston 2003).

The moratorium preventing exploitation of Antarctica’s resources will dissolve in

2048. At that time, only countries that were signatories to the Antarctic Treaty or have established research facilities on the continent will have a say in the next phase of

Antarctica’s use or exploitation (Leighton 2015). As of 2015, there were 53 signatories to the ATS (Secretariat of the Antarctic Treaty 2015).

Already, seven of the 12 countries that originally signed the ATS have staked claims to the continent, although other United Nations members fail to recognize these claims (Hemmings 2005; Jabor-Green & Nicol 2003). While most UN member nations believe that the High Seas extends to the shoreline of Antarctica, a few nations have asserted economic exclusion zones over parts of Antarctica (Hemmings 2005). The ATS was designed to prevent this free-for-all resource grab. If certain states feel they are being denied a share of resources, legally recognized or otherwise, this could create an unregulated scramble for a piece of Antarctica.

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Such competition may have already started. In 2011, at the 34th annual meeting of the ATS, Russia stated that it had goals to strengthen not only its bioprospecting capabilities on and around Antarctica, but to increase its natural resource exploration in the Southern Ocean. Twenty-nine countries now have research facilities on Antarctica, bringing the number on the continent to 82. Nor are developed countries the only ones that are attempting to secure a claim to the continent. China, Brazil, Ecuador, Chile,

Venezuela, and Argentina are among those countries that recently established or are in the process of establishing research bases in order to be at the table when Antarctica is carved up in 2048 (Leighton 2015). By the time that happens, the damage may have been done.

Among the many questions surrounding bioprospecting on and around Antarctica are: Does bioprospecting conflict with the ATS, or should it be banned from the continent? If allowed, who owns the genetic resources—the discovering party or the world? Who can access these genetic resources? If benefit sharing is enacted, who determines what is reasonable? What are the environmental and geopolitical ramifications of bioprospecting proceeding without a governing framework? Presently, the only rule governing bioprospecting in Antarctica is that there are no rules.

Much of the problem is due to legal conventions chasing technology. Like

UNCLOS, the ATS was conceived largely to regulate mineral and oil extraction.

Bioprospecting, specifically for microbes, was not on the radar in previous decades

(Hemmings 2005; Jabor-Green & Nicol 2003; Leary 2008b; Lohan & Johnston 2003).

The disparity between UNCLOS and the ATS, the two main documents governing

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Antarctica and the Southern Ocean, along with uncertainty about how the CBD applies to bioprospecting on Antarctica, if at all, has created a regulatory vacuum.

The international body is still undecided as to whether bioprospecting should be regulated by national or international laws (Rothwell 2005). Further, it is unclear what if any jurisdictional domain the CBD may have over bioprospecting in this part of the world

(Rothwell 2005). Such determinations rest on if Antarctica falls outside of any national domain or soveriegn jurisdiction, putting it out of reach of the CBD. One primary goal of the CBD was to regulate the transfer of genetic resources from provider to user. Given

Antarctica’s status, this part of the CBD does not appear to be applicable (Lohan &

Johnston 2005). As with bioprospecting and benefit sharing in the Area, the CBD offers no clear guidance for bioprospecting agreements on Antarctica. This goes back to the

CBD regulating only biological material that falls under a sovereign nation’s jurisdiction.

The Nagoya Protocol does not apply to Antarctica (Buck & Hamilton 2011).

The Antarctic Treaty System does not provide strict guidance for bioprospecting in Antarctica. Article III in the ATS calls for the free sharing of scientific knowledge derived from research on Antarctica. However, scientific knowledge is left undefined. It is not clear if the term includes bioprospecting research (Johnston & Lohan 2005).

Before legal constructions providing for bioprospecting around and on Antarctica can be adopted, it will be necessary to define exactly what bioprospecting is, at least in regards to Antarctica. Hemmings and Rogan-Finnemore (2005) point out the near impossibility of this. Bioprospecting grew out of scientific activities. It some respects it is indistinguishable. Yet it is also recognized as having both scientific and industrial goals.

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The ATS is predicated on the freedom of scientific research and collaboration, with a certain amount of mutually respected autonomy for science and its activities

(Hemmings & Rogan-Finnemore 2005). This leaves the door open for scientific principles to come under the sway of commerical interests.

The very nature of bioprospecting in both deep sea and Antarctic environments makes them enormously expensive ventures. As discussed, close associations have been formed between researchers and industry to offset these expenses. Graham (2005) has suggested that this leads to a tight and potentially cloudy intimacy between scientific and commerical interests.

Like deep sea bioprospecting, it can be difficult to delineate what is research bioprospecting and what is commercial bioprospecting. Graham (2005) points out that it is easier to erect a system that enforces the removal of a resource like oil, than it is to monitor the removal of a biological product potentially worth billions, when it can be smuggled out of Antarctica in a Petri dish.

The lack of a universally accepted and enforceable legal regime for intellectual property as it applies to bioprospecting on Antarctica is a critical missing link (Graham

2005). Unresolved are the processes governing patents when an Antarctic microbe is discovered by a country (Leary 2008a), and for benefit sharing disputes that may arise

(Johnston & Lohan 2005).

Yet even with all of these unresolved concerns, bioprospecting on Antarctica and in the Southern Ocean goes forward. Given the stakes for potential microbial bioprospecting and other resource extraction opportunities, this inability to resolve issues

181 may be intentional. That is, as many countries may see it, when it comes to their own national economic and political interests, the best regulation may be none at all.

Environmental damage to Antarctica and the Southern Ocean from unregulated bioprospecting is a real concern. For almost a quarter of a century bioprospecting has been underway in the Antarctic without regulation or legal guidelines, despite it being considered a sensitive environment. Some fear this might be leading to a “frontier mentality” (Graham 2005, 50).

There is good cause for concern. In addition to the growing number of tourists, researchers have caused documented damage to some degree. Japanese scientists researching a lake were accused of damaging it with radiological pollution; German researchers were believed to have trampled a sensitive moss bed near their research station; New Zealand scientists used fumaroles on Mt. Erebus as a barbecue pit; Chinese construction laborers were accused of using a penguin to play football; a French airstrip was allegedly constructed in a sensitive penguin environment; incidences of poaching toothfish by Spanish nationals have been reported (Graham 2005).

If an international legal regime for bioprospecting on Antarctica is left unresolved, it risks leaving the door open for mass corporate exploitation of biological resources (Hemmings & Rogan-Finnemore 2005). If a microbe of enormous pharmaceutical and commercial potential is discovered on Antarctica, extensive environmental damage may result before any guidelines or governing laws are in place.

As bioprospecting increases, and therefore the economic stakes of bioprospecting intensify, the belief that the Antarctic is considered fair game by all countries to perform

182 bioprospecting at will and without regulation, could make the continent hotly contested for reasons beyond microbes.

As natural resources are depleted on other continents, Antarctica could become too tempting to resist. This global commons could become a global contention. It will be difficult for science to remain disentangled from political tensions on Antarctica. As

Hemmings (2010) astutely pointed out, since territorial and ownership issues on

Antarctica are by nature political, there is little doubt science will be politicized.

Lack of a consensus on how to proceed with bioprospecting in Antarctica has the potential to go beyond scientific competition for exotic microbes. It could create geopolitical tensions among nations. China is rapidly increasing its funding for Antarctic bioprospecting. In 2012, China doubled its budget to $55 million. Still, this is much less than the estimated $300 million that the United States spends annually on its Antarctica bioprospecting research program. Although signatory countries to the Antarctic Treaty agreed not to extract Antarctica’s resources until 2048, there are ends around this through tourism, which is non-regulated, and geographical surveys (Robson 2013). Potentially, both could be used as fronts for less benign activities.

As time drags on without agreement, the harder it will be to create consensus.

Discovery of a commercially viable microbe by one country will create reasons to suspend further agreements as other countries scramble to lay claim to their own wonder microbe. Upon doing so, most countries will probably find it hard to relinquish potential wealth by constraining themselves through regulating bioprospecting. Antarctica will probably be regarded as a common heritage of mankind as long as it fails to produce any microbial wealth. That is highly unlikely.

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Bioprospecting in the Arctic

Issues in the Arctic regarding microbial bioprospecting are not any more resolved than those regarding Antarctica. Arctic bioprospecting has similar issues to its Antarctic counterpart, such as access; impact of scientific research on the environment; and sovereignty issues. Arctic microbial bioprospecting is generally carried out in territorial waters or state jurisdiction. Still, there is contention among states as to what constitutes territorial waters and who controls what waters.

The Arctic Council—a forum of countries including Canada, the United States,

Greenland, Finland, Iceland, the Russian Federation, and Sweden, charged with creating

Arctic policies—has provided no clear guidance on bioprospecting. Denmark and

Sweden have said they will not require PIC for genetic resources obtained from the wild, and therefore benefit sharing is not regulated. However, Greenland, which is controlled by Denmark, but autonomous, does regulate these activities. Iceland does not regulate bioprospecting except for that of microbes obtained from geothermal areas, although there is uncertainty about how geothermal areas are defined. Finland is still considering regulating bioprospecting in its Arctic areas (Leary 2008a).

Bioprospecting is underway in multiple Arctic regions. Greenland has come under attention from bioprospectors. Several Finnish and Spanish companies are in the early stages of exploring Greenland’s biodiversity. In the past decade Norway has aggressively stepped up bioprospecting activities in the Barents Sea. Iceland has seen its share of bioprospecting activities increase. Iceland straddles the Atlantic Mid-Oceanic Ridge where the seafloor is spreading. Diversa Corporation has reportedly secured rights to

184 commercially exploit all uses from a microbe discovered near an underwater hydrothermal vent along the Kolbeinsy Ridge (Lohan & Johnston 2003).

It is in Norway where the greatest potential for regional conflict exists due to bioprospecting. The Spitzbergen Archipelago, known as the Svalbard, is a possible point of contention should Norway decide to flex its territorial power in the face of Arctic bioprospecting competition. The Spitzbergen Treaty recognizes Norwegian control of the islands. But the treaty is unclear on how Norwegian sovereignty affects access and benefit sharing for foreign nationals engaged in bioprospecting, in regards to the CBD.

Nor is it resolved if Norway can extend control through the Continental Shelf and

Territorial Waters of its EEZ. Both Iceland and Russia have already contested Norway’s interpretation of the treaty in regards to fisheries (Leary 2008a).

The expansion into a new era of microbial bioprospecting may create a new shift in claims of biopiracy. There is a tendency to think that accusations of biopiracy falls on a developed-developing world axis. That may no longer be the case. Many people in

Norway have accused the pharmaceutical company Novartis of biopiracy. Sandimmum and Sandimmum Neoral are well-known drugs manufactured by Novartis. Together, these two drugs are believed to earn revenues of $1.2 billion annually. It so happens that these drugs were derived from a roadside plant without any benefit sharing agreement in place. Since this occurred in 1969, over 20 years before the CBD, Novartis feels it has no obligation to share revenues with Norway (Leary 2008a).

Chapter Summary

Bioprospecting on Antarctica and in the Southern Ocean is nowhere near being resolved. Bioprospecting is proceeding before any guidelines are in place. Unlike before,

185 when intellectual knowledge and benefit sharing was an issue, much more is at stake. Of all the new frontiers in microbial bioprospecting, Antarctica may prove to be the most contentious. The “Last Place on Earth” has the potential to become a flashpoint of global tensions unless bioprospecting guidelines are implemented.

If bioprospecting is allowed to proceed without regulation, there will be mission creep as countries compete to find ways to circumvent the ATS, and secure their own resources on Antarctica before it is snatched up. Ultimately, these tensions will not be just about microbial bioprospecting, but about resources that wars have been fought over: oil, gas, and minerals. All of this is instructive. Be it in the rainforest, the Arctic, the

Antarctic, a national park, or in the deep abyss of the ocean, bioprospecting for plants or microbial organsims will always be contentious.

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CHAPTER 14

MICROBIAL BIOPROSPECTING: A NEW ERA

Great Potential, Great Obstacles

Review

This thesis established that specific events over the past several hundred years encouraged a transition toward microbial bioprospecting. The events responsible for this development appear unconnected when examined individually, as prior literature has demonstrated. By examining these events over a several hundred-year scale, particularly from the 1800s on, this thesis has shown that each event acted as a successive link in the chain toward a new era in microbial bioprospecting.

The biopiracy of cinchona seeds left indigenous people embittered and suspicious of bioprospectors from developed countries. The biopiracy of cinchona served as a rallying cry for the promulgation of the CBD over 100 years later. Bioprospecting merged with science in the 1800s and set the foundation for drug synthesis in the following century. The new science of pharmacognosy based many laboratory tests on medicinal plants according to how those plants were used by indigenous peoples. Much of that literature came from early ethnobotanists in the field. By the end of the 1800s, ethnobotany became a recognized discipline. Ethnobotanists, for better or for worse, became forever linked to pharmaceutical bioprospecting.

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In the 20th century, plants fell in and out of favor as a source for pharmaceuticals.

Emerging biotechnologies contributed to these cyclical swings. During periods of renewed interest in plants, traditional knowledge often served as the basis for investigating bioactive compounds in plants. During the 1980s and 1990s, a perception arose that miracle drugs were being lost as the (Amazonian) rainforest was cleared.

Allegations of contemporary biopiracy garnered increasing attention. There was a desire to allow indigenous peoples to profit from their traditional medicinal knowledge. These factors merged with long-simmering animosity over past biopiracy to drive the adoption of the Convention on Biological Diversity.

The CBD was billed as a bridge that would allow indigenous peoples to profit from their traditional medicinal knowledge while pharmaceutical companies would tap that expertise to find leads for new drugs. This optimism was short lived. Many bioprospectors were stymied by the CBD’s inability to facilitate research. Developing countries became increasingly frustrated at the lack of revenue generated from benefit sharing agreements.

The CBD had been hailed as a new era for bioprospecting. Indeed, it was.

Researchers looked elsewhere and saw recent innovations in biotechnology had finally unlocked the door to the exploitation of microbial organisms. Although the CBD was later amended by the Nagoya Protocol, by then microbial bioprospecting was established.

Wrongly held perceptions served as an important link in the chain. The narrative that the traditional ethnobotanical knowledge of indigenous peoples was being exploited by pharmaceutical companies was a primary driver for promulgation of the CBD. Three case studies showed that this narrative is usually inaccurate.

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Even instances with a strong argument for biopiracy can be difficult to parse. The

San people in southern Africa may have the strongest case of biopiracy in over 100 years.

The Hoodia plant is endemic to a specific geographic area. It has not been transplanted to other countries. The San people are believed to have originated Hoodia’s use as an appetite suppressant. Even so, this example highlights typical concerns associated with designating ownership of TEK and benefit sharing. The San did share their knowledge with other groups. Since the San are largely without a central leadership, it raises the question of who can speak or negotiate on their behalf. Nor have the San received the substantial benefit sharing revenues originally envisioned by the early success of Hoodia.

This leads to a second important misperception: that the path from plant to successful pharmaceutical is an easy process. Two case examples showed that developing a successful drug from a plant is such a herculean enterprise that failure is the typical result.

The tenth chapter focused on the expansion of bioprospecting into a new era.

Microbial organisms known as extremophiles have generated a stir among researchers.

These bizarre microbes, dubbed extremozymes, inhabit environments once thought prohibitive to life. The anticipated potential of the enzymes or secondary metabolites contained in these microbes is so great, that researchers expect to develop not just revolutionary pharmaceuticals, but numerous applications for a variety of industrial sectors.

The eleventh, twelfth, and thirteenth chapters explored the environments that will be (and are) the next frontiers in microbial bioprospecting. For the first time America’s national parks are the focus of bioprospecting benefit sharing agreements. It was a

189 fumarole in Yellowstone National Park that gave rise to modern DNA technology. The

PCR technology based on a Yellowstone thermophile spurred the advancements in DNA technology that resulted in the first genetic sequencing of an organism in 1995. That in turn created a snowball effect of continuing innovations in genomic biotechnology that enabled the expansion into microbial bioprospecting (Bull 2004).

Unlike the bioprospecting of plants, this new era of microbial bioprospecting takes place in multiple geographies: terrestrial fumaroles, the abyssal depths of the ocean, and the frozen expanse of Antarctica. Formerly, plants and other terrestrial biological products served as the basis for many pharmaceuticals. Over the past twenty years, a biotechnology revolution has enabled bioprospecting to expand beyond its former historical limitations. Bioprospecting has moved from rainforest to fumaroles.

Lessons Learned and Concluding Thoughts

The expansion of bioprospecting into new geographical realms also enlarges the complications associated with bioprospecting. While the United States has laws in place that should largely protect its national parks from exploitation, laws governing bioprospecting in the deep ocean and on Antarctica are minimal. Here arises not only the potential for environmental and biodiversity damage, but the possibility for geopolitical tensions. The politics associated with microbial bioprospecting appear to be moving from a developed-developing world polarity to potential confrontations between developed militarized nations.

How bioprospecting is conducted in deep sea and Antarctic environments is strikingly different from that of terrestrial bioprospecting. Terrestrial bioprospecting projects can be conducted for a paltry sum when weighed against the massive costs of

190 microbial bioprospecting research in the Area or Antarctica. Both require investments that far exceed the financial capability of most private companies.

This has given rise to a new form of bioprospecting: that of the public-private consortia. Although such structures allow research to go forward, it also increases the chances for purely scientific research to be subsumed by commercial interests. In deep ocean environments or on Antarctica, where legal guidance is vague or absent, this could create issues that surpass the troublesome negotiations on traditional knowledge.

As bioprospecting transitions to microbes, sorting out complex issues of ownership will increase. If a medicinally important microbe is found in several different fumaroles across the world, who owns it? Or if a medicinally important microbe is discovered at only one deep sea fumarole, does the entire world claim ownership? These issues will make the discussions leading up to the CBD seem trivial. Bioprospecting has entered a new era, and so have the problems associated with it.

Aside from the political ramifications, it is appropriate to ponder the social implications of microbial bioprospecting. We have learned that many extremophiles possess a cellular structure that allows them to survive and thrive in the harshest conditions. If a microbe plucked from a fumarole in Yellowstone surrenders an enzyme that extends a human lifespan by rejuvenating cellular structure, it could have far reaching social implications.

The expansion to microbial organisms does represent a new era in bioprospecting.

The events discussed herein appeared unrelated but became linked over a long period. No one could have guessed that an astute observation by Jesuits priests, based upon traditional ethnobotanical knowledge of indigenous people, would become a vital

191 foundation for events that would transpire hundreds of years later. Nor was the expansion to microbial bioprospecting predictable. Early drug research based on plants in the 1800s jumped abruptly to synthetics in the next century. Yet as expectations went unfulfilled, interest swung back to plants and then away, as technology advanced in fitful stages.

The expansion to microbial bioprospecting arrived before the world recognized it.

The 1980 decision by the Supreme Court in Diamond v. Chakrabarty laid important legal ground work. The ability to patent a genetically-altered life form (Peterson 2001), and the biotechnology associated with the Human Genome Project (Hayden 2003), would have profound implications for microbial bioprospecting. The ink was barely dry on the CBD before the genetic sequence of Haemophilus influenzae was mapped and published three years later (Bull 2004).

Looking Forward

What Will Become of Plants?

A survey of microbial bioprospecting literature turns up the word “potential” in regards to microbes about as much as “biopiracy” is associated with bioprospecting from plants. Potential is key here. At one time plants had great potential. Then synthetic chemistry had great potential. Then plants reclaimed the mantle of potential, only to surrender it to microbial organisms. Waves of expectations followed by failure, followed by hope again, is symptomatic of the process of deriving pharmaceuticals from plants. It will remain so with microbial organisms.

Considering the opportunities that microbial organisms present, do plants have a future? Most certainly. As Miller (2011) points out, plants will maintain a prominent role in drug development from natural products. But they will have to share the limelight with

192 microbial organisms, or other biological sources. Lopez (2011) agrees that plants will not only continue to serve as a source for the basis of pharmaceuticals and nutraceuticals, but that research may increase.

While this thesis focused on microorganisms found in three geographical realms, useful microbes can be derived from other biological products—namely, fungi (Hunter-

Cevera 1998; Newman & Cragg 2012). Since the tropics contain a high diversity of microbial life (Hunter-Cevera 1998), it may allow developing countries to tap into the new era of microbial bioprospecting.

Yet this returns to the problems associated with the earlier disillusionment over lack of benefit sharing. Developing countries must understand the extremely low chance of success associated with pharmaceutical development. This is why some are recommending that benefit sharing should focus less on royalties that may come only after a decade or so—if ever—and instead emphasize attainable goals in the near future, like training, sharing technology, or disbursing milestone payments (Cragg, Katz,

Newman, & Rosenthal 2012).

Like deriving pharmaceuticals from plants, creating successful drugs from microbes will be frustrated by unforeseen obstacles. But microbial bioprospecting will proceed. Despite the great economic investment required, the myriad of potential uses for extremophiles and their enzymes is too tempting for biotechnology to resist. Whether it be the steaming fumaroles or bubbling mud pots in Yellowstone, a hydrothermal vent thousands of feet beneath the ocean surface, or the frigid ice of Antarctica, the expansion of bioprospecting from forest to fumaroles will ripple out to affect our lives—socially and politically—in ways we cannot imagine.

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The Future of Bioprospecting

So what is the future of bioprospecting? When all of Earth’s biological material has been thoroughly screened for bioactive compounds and there are conceivably no more new compounds to be identified, where will bioprospectors turn? Two words answer our question: look up.

Look up into the heavens at the small points of light. Specifically, asteroids, comets, and meteoroids swirling about in our solar system. Extraterrestrial bodies may hold the most extreme extremophiles in existence. Microorganisms may exist on Mars or in the icy waters of Jupiter’s moon Europa (Priscu & Christner 2004). Organic compounds have been recovered from meteorites, and research has shown that microbes can survive in space (Rothschild & Mancinelli 2001). Possibly, life was carried to Earth on a microbe by a meteroid that impacted the planet (Cavicchioli 2002).

As space and extraterrestrial bodies such as the moon are considered the common heritage of mankind (Rothwell 2005), the same issues seen here on Earth will no doubt follow bioprospectors as they reach out into space—the future era of bioprospecting.

APPENDIX A

FARNSWORTH’S LIST OF 47 MEDICINES FROM TROPICAL PLANTS

The following table shows 47 drugs (used in the USA) derived from 39 tropical plants. Adapted from Farnsworth’s original list of 121 drugs derived from 95 plant species, from both tropical and temperate zones. Adapted from Farnsworth & Soejarto 1989; Farnsworth & Soejarto 1991.

47 Drugs Derived from 39 Tropical Plants Species Drug Action / Clinical Use Species Drug Action / Clinical Use Vasicine Adhatoda vasica Oxytocic Erythroxylum coca Cocaine Local anesthetic (Peganine) Anamirta cocculus Picrotoxin Analeptic Gossypium species Gossypol Male contraceptive Anti-inflammatory; Ananas comosus Bromelain Lonchocarpus nicou Rotenone Piscicide Proteolytic Andrographis Andrographolide Bacillary dysentery Mucuna deeringiana L-Dopa Antiparkinsonism paniculata Andrographis Neoandrographolide Bacillary dysentery Nicotiana tabacum Nicotine Insecticide paniculata Ardisia japónica Bergenin Antitussive Ocotea glaziovii Glaziovine Antidepressant Areca catechu Arecoline Anthelminthic Pausinystalia yohimba Yohimbine Aphrodisiac Physostigma Physostigmine Cholinesterase Carica papaya Papain Proteolytic venenosum (Eserine) inhibitor Carica papaya L Chymopapain mucolytic Pilocarpusjaborandi Pilocarpine Parasympathomimetic Catharanthus roseus Vinblastine Antitumor Piper methysticum Kawain Tranquilizer Catharanthus roseus Vincristine Antitumor Quisqualis indica Quisqualic acid Anthelminthic 194

47 Drugs Derived from 39 Tropical Plants (continued) Species Drug Action / Clinical Use Species Drug Action / Clinical Use Antihypertensive; Centella asiatica Asiaticoside Vulnerary Rauvolfia canescens Deserpidine tranquilizer Cephaelis ipecacuanha Emetine Amoebicide; emetic Rauvolfia serpentina Ajmalicine Circulatory disorders Chondodendron tomentosum Tubocurarine Skeletal muscle relaxant Rauvolfia serpentina Rescinnamine Antihypertensive Cinchona ledgeriana Quinine Antiarrhythmic Antimalarial Rauvolfia serpentina Reserpine Antihypertensive Cinchona ledgeriana Quinidine Antipyretic Ricinus communis Castor oil Cathartic Cinnamomum camphora Campfior Rubefacient Rorippa indica Rorifone Antitussive Cissampelos pareira Cissampeline Skeletal muscle relaxant Simarouba glauca Glaucarubine Amoebicide Crotalaria sessiliflora Monocrotaline Antitumor Stevia rebaudiana Stevioside Sweetener Curcuma longa Curcumin Choleretic Stevia rebaudiana Rebaudioside A Sweetener Datura metel Scopolamine Sedative Strophanthus gratus Ouabain Cardiotonic Dioscorea species Diosgenin Source of female contraceptive Strychnos nux-vomica Strychnine CNS stimulant Dubosia myoporoides Atropine Anticholinergic Theobroma cacao Theobromine Diuretic Dubosia myoporoides Hyoscyamine Anticholinergic

195

APPENDIX B

DRUGS DERIVED FROM TRADITIONAL ETHNOBOTANICAL KNOWLEDGE

The following table shows 50 drugs derived from plants over approximately a 200-year period based on traditional ethnobotanical (medicinal) knowledge. From Balick & Cox, 1996.

50 Drugs Derived from Leads Based on Ethnobotanical Uses Drug Medical Use Plant Species Family Ajmaline Heart arrhythmia Rauvolfa spp. Apocynaceae Aspirin Analgesic, inflammation Filipendula ulmaria Rosaceae Atropine Ophthalmology Atropa belladonna Solanaceae Benzoin Oral disinfectant Styrax tonkinensis Styracaceae Caffeine Stimulant Camellia sinensis Theaceae Camphor Rheumatic pain Cinnamomum camphora Lauraceae Cascara Purgative Rhamnus purshiana Rhamnaceae Cocaine Ophthalmologic anaesthetic Erythroxylum coca Erythroxylaceae Codeine Analgesic, antitussive Papaver somniferum Papaveraceae Colchicine Gout Colchicum autumnale Liliaceae Demecolcine Leukemia, lymphomata Colchicum autumnale Liliaceae Deserpidine Hypertension Rauvolfia canescens Apocynaceae Dicoumarol Thrombosis Melilotus officinalis Fabaceae Digoxin & Digitoxin Atrial fibrillation Digitalis purpurea Scrophulariaceae Emetine Amoebic dysentery Cephaelis ipecachuanha Rubiaceae Ephedrine Bronchodilator Ephedra sinica Ephedraceae Eugenol Toothache Syzygium aromaticurn Myrtaceae 196

50 Drugs Derived from Leads Based on Ethnobotanical Uses (continued) Drug Medical Use Plant Species Family Gallotanins Hemorrhoid suppository Hamamelis virginiana Hamamelidaceae Hyoscyamine Anticholinergic Hyoscyamus niger Solanaceae Ipecac Emetic Cephaelis ipecacuanha Rubiaceae Ipratropium Bronchodilator Hyoscyamus niger Solanaceae Morphine Analgesic Papaver somniferum Papaveraceae Noscapine Antitussive Papaver somniferum Papaveraceae Papain Attenuates mucus Carica papaya Caricaceae Papaverine Antispasmodic Papaver somniferum Papaveraceae Physostigmine Glaucoma Physostigma venenosum Fabaceae Picrotoxin Barbiturate antidote Anamirta cocculus Menispermaceae Pilocarpine Glaucoma Pilocarpus jaborandi Rutaceae Podophyllotoxin Condylomata acuminata Podophyllum peltatum Berberidaceae Proscillaridin Cardiac malfunction Drimia maritima Liliaceae Protoveratrine Hypertension Veratrum album Liliaceae Pseudoephedrine Rhinitis Ephedra sinica Ephedraceae Psoralen Vitiligo Psoralea corylifolia Fabaceae Quinidine Cardiac arrhythmia Cinchona pubescens Rubiaceae

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50 Drugs Derived from Leads Based on Ethnobotanical Uses (continued) Drug Medical Use Plant Species Family Quinine Malaria prophylaxis Cinchona pubescens Rubiaceae Rescinnamine Hypertension Rauvoljia serpentina Apocynaceae Reserpine Hypertension Rauvolfia serpentina Apocynaceae Sennoside A,B Laxative Cassia angustifolia Caesalpiniaceae Scopolamine Motion sickness Datura stramonium Solanaceae Stigmasterol Steroidal precursor Physostigma venenosum Fabaceae Strophanthin Congestive heart failure Strophanthus gratus Apocynaceae Teniposide Bladder neoplasms Podophyllum peltatum Berberidaceae THC Antiemetic Cannabis sativa Cannabaceae Theophylline Diuretic, asthma Camellia sinensis Theaceae Toxiferine Surgery, relaxant Strychnos guianensis Loganiaceae Tubocurarine Muscle relaxant Chondrodendron tomentosum Menispermaceae Vinblastine Hodgkin’s disease Catharanthus roseus Apocynaceae Vincristine Pediatric leukemia Catharanthus roseus Apocynaceae Xanthotoxin Vitiligo Ammi majus Apiaceae 198

199

APPENDIX C

YELLOWSTONE NATIONAL PARK’S EXTREMOPHILE HABITATS

Top: Mud pots in Yellowstone harbor numerous extremophiles in their gooey super-heated muck. Photograph by Austin & Jones, 1987. Middle: Black Growler thermal steam vent (fumarole). From National Park Service, 2016; Photograph by Robinson, 1950. Bottom: An infrared image (left) and visible image (right) of Yellowstone’s Old Faithful geyser. Cooler colors indicate cooler temperatures. Warmer colors indicate higher temperatures. Courtesy of Cool Cosmos and NASA/JPL-Caltech, 2015. 200

APPENDIX D

DEEP OCEAN RIDGES AND THE MECHANICS OF HYDROTHERMAL VENTS.

Top: Map of mid-ocean ridges looking like a suture across the Earth’s ocean bottom. Colored areas indicate locations of known hydrothermal vent fields. From Tivey 2004, in Flores & Reysenbach, 2011. Bottom: Detailed geological mechanics of a deep sea hydrothermal vent, known as a black smoker. From PMEL-NOAA, 2015.

201

APPENDIX E

ANTARCTICA IS A NEW FRONTIER IN BIOPROSPECTING

Top: An ICE Sat image (May 2003) of Antarctica. Image from the CIA Factbook, courtesy of NASA/Goddard Space Flight Center & Scientific Visualization Studio, Canadian Space Agency, RADARSAT International Inc., 2015. Bottom: Map of Antarctica from Ingolfsson, 2004.

202

APPENDIX F:

ACRONYMS AND ABBREVIATIONS

ATS ...... Antarctic Treaty System CBD ...... Convention on Biological Diversity CHM ...... Common Heritage of Mankind COMPITCH ...... Consejo de Medicos y Parteras Indigenous Tradicionales de Chiapas CRADA ...... Cooperative Research and Development Agreement CSIR ...... (South African) Council for Scientific and Industrial Research ECOSUR ...... El Colegio de la Frontera Sur EEZ ...... Exclusive Economic Zone ICBG ...... International Cooperative Biodiversity Groups (Program) INBio...... La Asociacion Instituto Nacional de Biodiversidad ISA ...... International Seabed Authority MHV ...... Marine Hydrothermal Vent NCI ...... National Cancer Institute NGO ...... Non-Governmental Organization NIH ...... National Institutes of Health NPS ...... National Park Service PCR ...... Polymerase Chain Reaction PIC ...... Prior Informed Consent RAFI (ETC) ...... Rural Advancement Foundation International (aka ETC: Action Group on Erosion, Technology, and Concentration TEK ...... Traditional Ethnobotanical Knowledge TMK ...... Traditional Medicinal Knowledge UNCLOS ...... United Nations Convention on Law of the Seas YNP ...... Yellowstone National Park

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