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<I>Crematogaster</I> Ants Harriet L. Wilkes Honors College Honors Theses Florida Atlantic University Libraries Year Bioprospecting for useful compounds in the venom of Crematogaster ants Robert E. McCurdy Florida Atlantic University, This paper is posted at DigitalCommons@Florida Atlantic University. http://digitalcommons.fau.edu/wilkes theses/32 Bioprospecting for Useful Compounds in the Venom of Crematogaster Ants by Robert E. McCurdy A Thesis Submitted to the Faculty of The Wilkes Honors College in Partial Fulfillment of the Requirements for the Degree of Bachelor of Arts in Liberal Arts and Sciences with a Major Concentration in Environmental Science and a Minor Concentration in Chemistry Wilkes Honors College of Florida Atlantic University Jupiter, Florida May 2007 Bioprospecting for Useful Compounds in the Venom of Crematogaster Ants by Robert E. McCurdy This thesis was prepared under the direction of the candidate’s thesis advisor, Dr. James K. Wetterer, and has been approved by the members of his supervisory committee. It was submitted to the faculty of the Honors College and was accepted in partial fulfillment of the requirements for the degree of Bachelor of Arts in Liberal Arts and Sciences. SUPERVISORY COMMITTEE: ____________________________ Dr. James K. Wetterer ____________________________ Dr. Eugene Smith ______________________________ Dean, Wilkes Honors College ____________ Date ii ACKNOWLEDGEMENTS First of all, I want to thank everyone who contributed to my education. This includes everyone from family, teachers, professors, friends and anyone else who helped get where I am now. All the little or big things that helped me along the way will not go unforgotten. We are all products of our environments and I like to believe that I am a shining example of what an excellent education provides. Next is my family. You guys have been so great to me. Each one of you has contributed your part to being there when I needed you. Through thick and thin, you were always there for me. Mom, I don’t think I can ever say enough to express my gratitude and love for you. Thanks for believing in me. Paul, you have pushed me and provided me with a role model like no other. Most of all, thanks for being my best friend. Live it up in NZ and study hard. I’ll see you when I get there. Sarah, I love you for all you are; even when we don’t see eye to eye on everything. Congratulations on being the newest Dr. McCurdy. Alice, you bring a light to all of our lives like no other. Keep it up. Last, but not least, is my deceased father. Your memory is with me everyday. Thanks for showing me how wonderful life is, especially around the ocean. To the faculty of the Honors College, it was a real pleasure to learn with all of you. Most especially, I would like to recognize Dr. Wetterer, Dr. Smith, Dr. O’Brien, and Dr. Moore. Thanks for keeping my interest in the sciences and the environment alive. Working with each one of you on a personal level has meant a lot to me. Mahalo. iii ABSTRACT Author: Robert E. McCurdy Title: Bioprospecting for Useful Compounds in the Venom of Crematogaster Ants Institution: Wilkes Honors College of Florida Atlantic University Thesis Advisor: Dr. James K. Wetterer Degree: Bachelor of Arts in Liberal Arts and Sciences Concentration: Environmental Science and Chemistry Year: 2007 Bioprospecting, the search for useful compounds in nature, has led to the discovery of many important pharmaceuticals. Most current bioprospecting efforts work with chemicals derived from marine invertebrates and terrestrial plants. I looked for useful compounds in a relatively unstudied source, the venom of Crematogaster ants, using Gas Chromatography-Mass Spectroscopy (GC-MS). Further studies can more accurately identify these chemicals using High Pressure Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR). iv TABLE OF CONTENTS Introduction………………………………………………………………………1 Methods…………………………………………………………………………...7 2.1 Study Site.…………………………………………………………………7 2.2 Laboratory…………………………………………………………………9 2.3 Gas Chromatography-Mass Spectroscopy………………………………...9 2.31 Gas Chromatography (GC)……………………………………………..10 2.32 Mass Spectroscopy (MS)……………………………………………….10 Results…………………………………………………………………………...12 Discussion…………………………………………………………………….…14 Works Cited…………………………………………………………………….16 v TABLE OF FIGURES FIGURES Figure 1 Crematogaster sp. ………………………………………………………...4 Figure 2 Land use map of Abacoa…………………………………………………..8 Figure 3 GC scan of Crematogaster venom……………………………………….13 vi INTRODUCTION The Secretariat of the Convention on Biological Diversity (CBD) has defined bioprospecting as “the exploration of biodiversity for commercially valuable genetic and biochemical resources” (Arico and Salpin 2005). Its main goal is to discover compounds that exist in nature with beneficial properties that can be used by humans. Once found and identified, the genetic or biochemical resources can be applied in many different fields, of which drug research is the most notable. Bioprospecting is a field that has received much interest lately as scientists seek out new approaches to curing disease. While drug companies dedicate valuable time and resources uncovering the pathologies of diseases so as to design new drugs to counter the effects, there may be a compound already in existence that an organism has been using for some time for this particular purpose. Therefore, research dealing with the discovery and properties of novel compounds in nature warrants more attention. What constitutes bioprospecting in international law is not yet undefined. A few countries have defined it in laws governing biodiversity but diverge in what steps to bringing a product to market actually encompass bioprospecting. To simplify matters, Arico and Salpin (2005) listed the steps: • Systematic search, collection, gathering or sampling of biological resources for purposes of commercial or industrial exploitation; • Screening, isolation, characterization of commercially useful compounds; • Testing and trials; and 1 • Further application and development of the compounds for commercial purposes, including large-scale collection, development of mass culture techniques, and conduct of trials for approved commercial sale. Sought after compounds could be identified by indiscriminately testing natural compounds from any organism, but chances of getting a promising candidate compound could be daunting. Instead, scientists begin their searches by seeking out organisms that have displayed desirable attributes. In the case of the rosy periwinkle (Catharanaus roseus), generations of people had been using this flower to treat diabetes in Madagascar (Gept 2004). When researchers took a closer look in the 1950s, they found a compound, which is later marketed as the drug Vincristine that helped cure thousands of childhood leukemia cases (Gept 2004). Here is an example of local knowledge of the environment leading to a treatment whose benefit to humanity is practically immeasurable. With so many potential compounds and organisms out there, how should scientists proceed with bioprospecting efforts. Like in the case of the rose periwinkle, traditional knowledge of plants can prove to be helpful. But in today’s modern societies, intimate knowledge of natural remedies have been lost; so instead, science should turn to organisms that demonstrate unique chemical solutions to environmental pressures. For this study, the unique properties of ant venom are investigated further. Bioactive compounds from ants are exploited by a variety of other species in nature. Many birds rub ants on their feathers, a behavior called “anting,” apparently to help control parasites (Revis and Waller 2004). The chemicals found in ants are thought to retard plumage deterioration by helping control microbe and fungal growth. Anting 2 creates better fitness for the birds, as well as making them more attractive to potential mates. In another case, the skin of certain poisonous frogs and toads contain poisonous compounds known as pumiliotoxins, apparently sequestered from ants and other arthropods in their diets (Jones et al. 1999). The frogs are able to utilize the chemicals created by the ants for defense for their own purpose of avoiding predation. Humans have only recently begun to tap into the potential of ant venoms. One research group has focused on engineering a more potent baculovirus for pest control on crops (Szolajska et al. 2004). Natural baculoviruses have proven to be effective, but slow acting insecticides. The lag time between infection and death has ruled out baculoviruses as a viable control method because significant damage can be inflicted on the crops before the insects die. By engineering the genetic material coding for poneratoxin, a neurotoxin from the ant Paraponera clavata, into the baculoviruses, the viruses kill the pests at a much quicker rate (Szolajska et al. 2004). The group has shown that the baculoviruses with the recombinant DNA are both a quick-acting and wide spectrum insecticide suitable for commercial use. These instances represent how chemicals from ant venom can be utilized for a multitude of applications. They also show that further studies can uncover even more useful compounds from the potent mix of chemicals that ants use to protect themselves from their enemies. Ant venoms are shown to be more structurally diverse than both bee and wasp venoms (Blum 1992). They have also been shown to have many biochemical pathways that are notable. As Blum (1992) reported: 3 The poison gland products of selected ant venoms are cytotoxic
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