1080 Chemical Facts by Stuart Campbell

1080: its chemistry, and the role of micro-organisms in its break down ML Jarman Introduction. It has been stated repeatedly to the NZ public that the current pest control regime, based on aerial use of manufactured 1080, is not harmful to the physical environment, and every living organism within that environment, because 1080: • • •

occurs in many plants and is therefore a `natural' chemical; `breaks down' into harmless chemicals (eg `salt and vinegar'); and is de-fluorinated and rendered non-toxic, by `commonly occurring' soil micro-organisms.

We contend that these emotive statements lack real scientific basis and that there are flaws, inaccuracies and unknowns in the whole subject of the chemistry of 1080 and its microbial de-fluorination, which make it misleading to the NZ public and dangerous to the NZ environment. To challenge the statements, we pose the following questions and try to find answers from our own knowledge backed up by information in scientific books, reports and publications. These questions have never been satisfactorily answered by the responsible authorities when we pose them in the consultation process. Questions 1. What is manufactured 1080? 2. Is there a natural form of manufactured 1080? 3. What is the chemical structure (what the molecule looks like) of manufactured 1080? 4. How is 1080 manufactured? 5. What is the chemical structure of potassium monofluoroacetate? 6. How and why does a plant manufacture monofluoroacetate? 7. Why is potassium monofluoroacetate not manufactured instead of the sodium version? 8. What is monofluoroacetate? 9. Why is monofluoroacetate so toxic? 10. How common are the plants that contain monofluoroacetate? 11. What can manufactured 1080 "break down" into? 12. What does de-fluorination mean? 13. Which are the `commonly occurring micro-organisms' that can de-fluorinate manufactured 1080 and how widespread and common are they? 14. What are the products of de-fluorination? 15. Where are the products of de-fluorination? 16. What are the factors controlling microbial activity?

Questions and Answers

1. What is manufactured 1080? The name `Compound 1080' is used for a relatively complex, manufactured chemical compound which can be stored in its anhydrous (dry) form as a crystalline powder. In its sodium salt form (CH2FCOONa) it is called sodium monofluoroacetate. 1,2,3 1080, sodium monofluoroacetate, sodium fluoroacetate and even sodium fluoroacetic acid, are some of the names that have been used for the same manufactured compound. 3 There are 21 synonyms used throughout the world .3 The chemical formula is CH2FCOONa. This is the chemical shorthand for the compound. It contains two carbon atoms, two hydrogen atoms, one fluorine atom (mono meaning one), one sodium atom, and two oxygen atoms in every molecule of the compound (Notice there is not a chlorine atom in sight.) 1,2 Ref: 1. Eisler R (1995) Sodium Monofluoroacetate (1080): Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review. Published by the U.S. Dept. of the Interior as Biological Report 27, Contaminant Hazard Reviews Report 30 February 1995 2. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 3. Norris WR (2001). Sodium fluoroacetate. In: International Programme on Chemical Safety. Poisons Information Monograph 494. Chemical. New Zealand National Poisons Centre, Box 913, Dunedin, New Zealand. Peer Review Update: Awang R, Besbelli N, Caldas, LQA; 17th. September 2001, Edinburgh Section 9 updated November 1, 2001 Dr WA Temple, Dunedin Final review November 2001 Penang Conclusion: Sodium monofluoroacetate is the chemical name which accurately describes the structure of manufactured 1080 used for pest control in New Zealand, and is the chemical name which should always be used in scientific publications and popular reports issued by agencies in New Zealand. It is sloppy and misleading to do otherwise. We are currently analysing a bibliography of more than 700 publications, reports, web site accounts, popular articles etc, that we have compiled, to see how prevalent this practice is, and how it may have affected the accuracy and credibility of the particular report.

2. Is there a natural form of manufactured. 1080? There is no natural form of manufactured 1080 (sodium monofluoroacetate). The chemical found in Dichapetalum cymosum (gifblaar = poisonous leaf) in South Africa is reported to be potassium fluoroacetate (CH2FCOOK)'. Other references to the same plant identify the toxic `principle' as merely `fluoroacetate'.2 Dichapetalum toxicarium (Chailletia toxicaria, Don) is a shrub occurring in Sierra Leone, West Africa, which produces a hard and woody fruit, extremely toxic to warm-blooded animals. `On the basis of nuclear magnetic resonance, infra-red spectroscopy and ozonolysis, the toxic ‘principle’ was found to © MLJarman CMB 90 Owhango 3990 via Taumarunui, NZ

be 18-fluoro-cis-9-octadecenoic acid (w-fluoro-oleic acid), or F(CH2)8CH=CH(CH2)7C00.3,4 Published research into the toxic component of some species of the Australian plant genus, Gastrolobium, and its environmental persistence does not call the toxic component 1080, or sodium monofluoroacetate. It refers to it as the toxic fluoroacetate (CH3FCOO-) part of the chemical. 5 Other researchers use the same terminology and make the same correct distinction. 6, 7, 8 Press and popular articles, written for public information and reassurance, do not make the distinction. They also use emotive language. 9.10 References: 1. Myers AG et al (2001) Journal of the American Chemical Society. 123:7207 "The identification of potassium fluoroacetate as the toxic principle of the South African plant Dichapetalum cymosum in 1943, by JS Marais, is often regarded as an important early discovery that directed attention to the potential of fluorine substitution to profoundly influence the biological activity of organic molecules." 2. Grobbelaar N & Meyer JJM 1989. Fluoroacetate production by Dichapetalum cymosum (gifblaar). Journal of Plant Physiology 135:550-553. 3. Pattison FLM & Dear REA (1961) Synthesis of the Toxic Principle of Dichapetalum toxicarium (18Fluoro-cis-9-Octadecenoic Acid). Nature (192) 1284-1285). Department of Chemistry, University of Western Ontario, London, Ontario. 4. Goldman P (1965). The enzymatic cleavage of the carbon-fluorine bond in fluoroacetate. The Journal of Biological Chemistry 240(8):3434-3439 5. Bong CL, Cole ALJ & Walker (1979). Effect of sodium monofluoroacetate (compound 1080) on soil microflora. Soil Biology and Biochemistry. 6. King DR, Kirkpatrick WE, Wong DH & Kinnear JE (1994). Degradation of 1080 in Australian soils. Proceedings of the Science Workshop on 1080. Royal Society of NZ, Miscellaneous Series 28 7. Twigg LE & King DR (1991) The impact of fluoroacetate bearing vegetation on Native Australian fauna: a review. Oikos 61: 412-430 8 Eason CT, Gooneratne R & Rammell CG (1994). A review of the toxicokinetics and toxicodynamics of sodium monofluoroacetate in animals. Proceedings of the Science Workshop on 1080. Royal Society of NZ, Miscellaneous Series 28. "It is generally agreed that fluoroacetate (whether naturally occurring in plants or in the form of 1080)..." 9 Bieleski R (2002). The 1080 plant. Friends of the Auckland Regional Botanic Gardens. Newsletter June 2002 "To those interested in plants, the most fascinating thing about 1080 is that it is a natural product ....produced by a number of plant species ...chemists twigged that these plants contained heaps of fluoroacetate - that is 1080.... What all this means is that in using 1080, we are not putting a new poison into the environment, we are increasing the range and level of what is a natural occurrence. Thus the risks of putting synthetic 1080 into the environment have to be read in that light" 10. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 "Perhaps the most surprising fact about 1080 poison is that it is a natural product. It occurs in significant concentrations in a number of plants, most notably the South African gifblaar plant and at least forty Australian plant species. In fact, the toxicity of sodium monofluoroacetate was first recognised in the 1830's when settlers in the Transvaal noticed their cattle dying after having munched on the aforementioned gifblaar plant, although the exact chemical species responsible for the poisoning was not identified until over one hundred years later. . Now if only we could get those bloody opossums to develop a taste for the gifblaar plant..." © MLJarman CMB 90 Owhango 3990 Via Taumarunui NZ

Conclusion: There has been ambiguity in the use of the statement that manufactured 1080 is the same chemical as a naturally occurring compound, which has misled the public. This needs to be acknowledged to the public.

3. What is the chemical structure (what the molecule looks like) of manufactured 1080? Sodium monofluoroacetate is a manufactured carbon or organic compound. Carbon chemistry (organic) is different to that of all the other elements, which are inorganic. This is because of the number of electrons which it has in its outer valency shell (the electrons which control what it can form compounds with). It does not form carbon ions (atoms with electrons either added or taken away and thus carrying positive or negative charge), but shares the four outer electrons with elements it combines with. They in turn have one electron to contribute. Eight electrons represent a stable molecule. Each carbon atom thus forms 4 bonds represented by the lines drawn out in the formula structure below, when it is in combination with other elements. Each bond shares two electrons (subatomic particles which are essential for forming bonds between elements) between the carbon atom and the atom of the element it is joined to. In the structure drawn out below, the first carbon atom forms single bonds with hydrogen (two of them) and fluorine and the adjacent carbon atom. That gives you the 4 bonds. The second carbon atom forms a single bond with the adjacent carbon atom, and a single bond with one of the oxygen atoms, and a double bond with the other oxygen atom. This gives you 4 bonds. Oxygen has two electrons to contribute (because of its outer valency shell electron compliment) and thus needs to either form double bonds with carbon, or join onto another inorganic element atom in a loose ionic bond, shown as the dashed line between 0 and Na in the diagram. This is the only `ionic' type bond in inactive 1080. 1,2,3 H 0- -- -- --Na+ ⁄ ⁄ H―C―C ⁄ ⁄⁄ F 0

References: 1. Eisler R (1995) Sodium Monofluoroacetate (1080): Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review. Published by the U.S. Dept of the Interior as Biological Report 27, Contaminant Hazard Reviews Report 30 February 1995 2. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 3. Bennett SM (2002) Sodium monofluoroacetate (1080). http//the piedpiper.co.uk/this(j).htm Conclusion: 1. The structure of manufactured 1080 (sodium monofluoroacetate) is that shown above. The structure of the synonym sodium fluoroacetate is assumed to be the same – but it © MLJarman CMB 90 Owhango 3990 via Taumarunui NZ

COULD be sodium di-fluoroacetate, or sodium tri-fluoroacetate, by replacing one (CHF2COONa) or two (CF3COONa) of the hydrogen atoms with fluoride atoms. The precision of the term sodium monofluoroacetate is very important. 2. The synonym sodium fluoroacetic acid (CFH3COOHNa) cannot exist. There would be too many bonds on both carbon atoms. This is again misleading to most members of the public and irrelevant to many scientists without strong Chemistry background.

4. How is 1080 manufactured? Presumably, the chemical is manufactured under conditions of high temperatures, low pressure — or with the use of catalysts. The manufactured 1080 (sodium monofluoroacetate) used in New Zealand comes from Tull Factory in the USA. In a report which minimises the risks of health effects from eating contaminated ducks, the temperatures at which manufactured 1080 can disintegrate are given as being between 100 and 200° 1 . Cooking fixes it. An earlier report puts the temperature for sodium fusion to the other constituent elements to be 500°C 2 i.e. it takes very high temperatures to get sodium to join onto the other elements in manufactured 1080? It is manufactured and exported as a white anhydrous crystalline powder, and the baits are prepared in New Zealand. 1 Temple & Edwards in: Eisler R (1995) Sodium Monofluoroacetate (1080): Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review. Published by the U.S. Dept. of the Interior as Biological Report 27, Contaminant Hazard Reviews Report 30 February 1995. pps 26-27.' ....Oven-baking or grilling at temperatures of greater than 200°C causes break down of 1080' 2 Goldman P (1965). The enzymatic cleavage of the carbon-fluorine bond in fluoroacetate. The Journal of Biological Chemistry 240(8):3434-3439 Conclusion: The answer to this question is still being sought after. Tull Factory owners are unlikely to advertise their chemical process!

5. What is the chemical structure of potassium monofluoroacetate? The structure of the naturally occurring potassium monofluoroacetate is CH2F000K, with potassium (K) atom replacing the sodium (Na) atom in sodium monofluoroacetate.

6. How and why does a plant manufacture monofluoroacetate? The biological processes which produce chemicals in situ are complex and not fully understood. An over simplistic model is that the leaves of plants are the "factories" and they obtain their raw materials from the soil. But many of the plants which produce monofluoroacetate and which have been analysed for it, grow in soils which are low in fluoro-minerals (fluoride ions). ' Studies of plant manufactured monofluoroacetate show the site of the toxin in the plant (leaf, roots, shoots, fruits) varying from species to species 2, perhaps evolved in response to the way in which animals harvest the plant products?3 1 Hall RJ (1972). The distribution of organic fluorine in some toxic tropical plants. New Phytologist 71: 855-871. ..None of the plant species examined ...from Africa, Australia and South America ... grew in soils which were high in fluoro-minerals; on the contrary some of the plants grew in soils with © MLJarman CMB 90 Owhango 3990 via Taumarunui, NZ

exceptionally low levels of fluorine... 2 Ward PF & Huskisson NS (1960). The metabolism of fluoroacetate by plants. Biochem J. 113(2): 9p. 3 DR King, AJ Oliver and RJ Mead ( ). The adaptation of some western Australian mammals to food plants containing fluoroacetate. Australian Journal of Zoology 26(4) 699 — 712... The tolerance to fluoroacetate of mammals from the south-west of Western Australia is unusually high Others from eastern Australia are much more susceptible. This tolerance appears to be an adaptation to the presence of mono-fluoroacetate in many species of the plant genera Gastrolobium and Oxylobium, which occur within the range of these mammals in Western Australia. Conclusion: There is no simple answer to this question. It needs further investigation, both in the literature and experimentally. This should be acknowledged to the public.

7. Why is potassium monofluoroacetate not manufactured instead of the sodium version? Presumably it is more difficult to manufacture, and therefore more costly to produce?

8. What is monofluoroacetate? (Fluoroacetate) It is the monofluoroacetate ion (CH2FCOO) part of the sodium monofluoroacetate (1080) molecule. This means it is the group of atoms carrying a negative charge, with the weak ionic bond between it and sodium (Na)+ referred to in Question 3. The structure is shown in Figure1 on the right hand side.1

Figure 1 H O ⁄ ⁄⁄ H―C―C ⁄ ⁄ H Oacetate ion

H O ⁄ ⁄⁄ H―C―C ⁄ ⁄ F Omonofluoroacetate ion

It is the monofluoroacetate part of the manufactured (1080) sodium monofluoroacetate chemical molecule, or that derived from natural plants, which is the toxic part of the chemical i.e. the monofluoroacetate ion.2 References 1. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 2 Eason CT & Frampton (1991). Acute toxicity of sodium monofluoroacetate (1080) bait to feral cats. Wildlife Research 18(445-449) Conclusion: Regardless of the source of the monofluoroacetate molecule, it is the component of 1080 which causes death.

9. Why is monofluoroacetate (fluoroacetate) so toxic? Blackman (2002) states "In terms of toxicity, the sodium ion (Na+) is innocuous and it's the monofluoroacetate ion (CH2F000") that's the nasty part. And it's nasty primarily because of its shape. © ML Jarman CMB 90 Owhango 3990 via Taumarunui, NZ

Like many poisons, the monofluoroacetate ion closely resembles a non-toxic species that is essential for the life of the organism in question. In this case, that essential species is the acetate ion, the structure of which is also shown in Figure 1. Many living organisms utilise the acetate ion in the Krebs cycle. This is a series of chemical processes in which acetate is ultimately converted to carbon dioxide, resulting in the production of energy. The chemical structures of the acetate and monofluoroacetate ions are so similar that those organisms dependent on absorbing and utilising acetate ions will also absorb monofluoroacetate ions, thinking that they're acetate ions — big mistake! One of the many processes in the Krebs cycle involves the conversion of acetate ion to citrate ion (Figure 2). The organism will likewise convert the structurally similar monofluoroacetate ion to the monofluorocitrate ion (Figure 2), but in doing so, it makes a fatal mistake. The monofluorocitrate ion is highly toxic because it immobilises an enzyme necessary for the next step of the Krebs cycle. The cycle thus stops at this point, resulting in a build-up of citrate ion in the organism and this, coupled with the loss of energy derived from the Krebs cycle leads to death. So you can see that the monofluoroacetate ion isn't toxic per se — it's what happens to it once it is ingested that results in its toxicity." 1. H H

⁄ O H O H O ⁄⁄ ⁄ ⁄ ⁄ ⁄⁄ C―C―C―C―C ⁄ ⁄ ⁄ ⁄ ⁄ O- H C H O⁄ ⁄⁄ O- O citrate ion

⁄ O H O H O ⁄ ⁄ ⁄ ⁄ ⁄ ⁄⁄ C―C―C―C―C ⁄ ⁄ ⁄ ⁄ ⁄ O- F C H O⁄ ⁄⁄ O- O monofluorocitrate ion

The toxicity of monofluoroacetate (whether in manufactured 1080 or the natural form)2 is due to this unique ability of the fluoroacetate part of the molecule to enter the metabolic machinery of a cell and tie up the enzymes aconitase and succinate dehydrogenase in the Kreb's (or Tri-Carboxylic Acid) cycle. This blockage results in the prevention of energy production in the cell and the accumulation of citrate which ultimately leads to cellular damage and death. References: 1. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 2. Eason CT & Frampton (1991). Acute toxicity of sodium monofluoroacetate (1080) bait to feral cats. Wildlife Research 18(445-449) Conclusion: Fluoroacetate is a unique poison. It is much more toxic than other fluoride compounds because of its ability to fit such a vulnerable niche in cellular metabolism and indeed because of the tenacity of the fluorine-acetate bond. 2 © MLJarman CMB 90 Owhango 3990 via Taumarunui, NZ

10. How common are the plants that contain monofluoroacetate? Plants reported to contain fluoroacetate and the regions where they come from are listed below. It is important to make the distinction between plant species, plant genera and plant families. For example, Gastrolobium is a Genus (Group) belonging to the Fabaceae Plant Family. Gastrolobium grandiflorum is a particular species within the Genus Gastrolobium. Plant Families and Genera have a distribution pattern consistent with the breakdown of Gondwanaland, and it is not surprising to find the same Plant Families and Genera in tropical Africa, Australia and Latin America. Individual species are not shared between the continents. The geological history of these three continents is also ancient. Soil chemistry is also a complex topic, but it could be assumed that the raw materials of the plant produced toxins are available to plants because of their soil chemical composition which reflects the soil origins. Plant taxonomy (naming of plants and establishing their affinities) is also a complex topic and constantly being revised. Scientific accounts, in reports and publications cite the actual plant species. Anonymous accounts written for agencies to inform and reassure the public do not.1,2 Gastrolobium species (Fabaceae Family) Gastrolobium grandiflorum (Fabaceae Family) Acacia georgina Dichapetalum cymosum (gifblaar = poisonous leaf) (Dichapetalaceae Family) Dichapetalum toxicarium (Chailletia toxicaria, Don) (Dichapetalaceae Family) Oxylobium parviflonum var. Revolutum C.A.Gardner (taxonomic synonym of Gastrolobium melanocarpum G. Chandler & Crisp. Palicourea margravii (Rosaceae family)

South Western part of Western Australia 1,2,3,4,5 Queensland, Australia 1,8 Queensland, Australia 1,8,9 South Africa 6,8 Sierra Leone , West Africa 7,8 Australia 8,10

Brazil 8,11

References. I. McKenzie R (Dr) BVScMVScDVSc. Australian Native Poisonous Plants. Senior Pathologist with the Animal Research Division of the Queensland Department of Primary Industries at Yeerongpilly, Brisbane. From hhtp://farrer.csu.edu.au/ASGAP/APOL7/sep97-4.html 2. Bong CL, Cole ALJ & Walker (1979). Effect of sodium monofluoroacetate (compound 1080) on soil microflora. Soil Biology and Biochemistry. 3. King DR, Kirkpatrick WE, Wong DH & Kinnear JE (1994). Degradation of 1080 in Australian soils. Proceedings s of the Science Workshop on 1080. Royal Society of NZ, Miscellaneous Series 28 4. Twigg LE & King DR (1991) The impact of fluoroacetate bearing vegetation on Native Australian fauna: a review. Oikos 61: 412-430 5 Eason CT, Gooneratne R & Rammell CG (1994). A review of the toxicokinetics and toxicodynamics of sodium monofluoroacetate in animals. Proceedings of the Science Workshop on 1080. Royal Society of NZ, Miscellaneous Series 28. "It is generally agreed that fluoroacetate (whether naturally occurring in plants or in the form of 1080...” © MLJarman CMB 90 Owhango 3990 via Taumarunui,

NZ 6. Grobbelaar N & Meyer JJM 1989. Fluoroacetate production by Dichapetalum cymosum (gifblaar). Journal of Plant Physiology 135:550-553. 7 Pattison FLM & Dear REA (1961). Synthesis of the Toxic Principle of Dichapetalum toxicarium (18Fluoro-cis-9-Oetadecenoic Acid). Nature (192) 1284-1285). Department of Chemistry, University of Western Ontario, London, Ontario. 8 Anonymous (2001) Reference Set: Fluoroacetate. PFPC 9 Oelrichs PB, McEwan T (1962). The toxic principle of Acacia georginae. Queensland Journal of Agricultural Science 19(1):1-16. 10 Chandler et al., Austral.Syst.Bot.15:646(2002) 11 Harper DB &O'Hagan D(1994)Fluorinated natural products. Natural Products Report 11:123-133 12. Anonymous DoC website — pests "Although 1080 is a manufactured compound, the active ingredient in 1080, fluoroacetate, is a natural plant toxin found in a more than 50 plants (including the tea plant) in South Africa, South America and Australia The toxin evolved as a deterrent to browsing animals." Conclusions: There is ambiguity and error contained in the statements: (1) that many plants produce sodium monofluoroacetate. Naturally occurring products are either monofluoroacetate or a completely different chemical; and (2) that they are common and widespread. In the initial search for publications citing the plant species, the above are the ones that have come to light. This search is ongoing. There are no New Zealand examples. These plants also represent a very small percentage of the total flora in each of the regions mentioned above. (These percentages are being checked up on.) They occur in tropical regions growing on soils of ancient origins where there is herbivory.

11. What chemicals can manufactured 1080 break down into and how? The carbon-carbon bond in the sodium mono-fluoroacetate molecule is less stable (weaker) than the carbon-fluorine bond, and so over time and with hydration (adding H20), sunlight can break the bond down, and the molecule can degrade into sodium bicarbonate (NaHCO3 ) and methyl fluoride (CH3F). This could happen if the molecules are lying for some time in surface water in sunlight. How likely is this to be the situation in a forest? If the molecules have entered streams or ground water, they are also unlikely to break down via sunlight. The chemical equation for this possible break down, if it occurs, is: CH2FCOONa + H2O →NaHCO3 + CH3F with all elements and their symbols accounted for. Methyl fluoride is a volatile fluoro-hydrocarbon which rots the ozone layer and contributes to the ozone hole! (Now — there is an emotive statement!) Sodium bicarbonate is a metal salt (common name baking soda). Metal salt is the general scientific term for a positively charged metal ion (Na+) joined to a negatively charged ion of an element, or a radicle (group of elements behaving like an element) - in this case the bicarbonate ion (HCO3). It is this term (metal salt) which may have contributed to the `salt and vinegar' story. ' © MLJarman CMB 90 Owhango 3990 via Taumarunui, NZ

The salt and vinegar story. Some claim that hydration (adding water, as in rain) spontaneously causes the dissociation of the fluorine and acetate part of the 1080 molecule. This in turn has led to the `myth', often and recently repeated that it breaks down into harmless by-products, namely "salt and vinegar" 1,2 Salt (or common table salt) is sodium chloride (NaCI). Vinegar (or acetic acid) is CH3COOH. Sodium monofluoroacetate cannot "break down" into salt and vinegar. It does not contain the element Chlorine (Cl) so cannot form table salt. Vinegar is not a product of the degradation of sodium monofluoroacetate. Figure 1 shows the structure of an acetate ion. 3 Adding one hydrogen atom to it could indeed produce CH3COOH (acetic acid or vinegar). However, water (H20), has two hydrogen atoms and one oxygen one. What happens to the other hydrogen atom and the oxygen one? You cannot rip off part of a molecule unless all components of both chemicals (acetate ion and water) have been accounted for. This is impossible anyway, because the acetate ion is not the negative ion present in 1080. It is the monofluoroacetate ion which is present. Simply adding H2O to monofluoroacetate (CH2FCOO-) does not happen, and if it could, it cannot produce vinegar. There simply are not the right number of atoms present. When water is added to sodium monofluoroacetate it "dissolves" but does not lose its integrity. It is diluted as the molecules are spread out in water (like dissolving coffee in water), but the molecules remain as sodium monofluoroacetate molecules.

Figure 1 H O ⁄ ⁄⁄ H―C―C ⁄ ⁄ H Oacetate ion

H O ⁄ ⁄⁄ H―C―C ⁄ ⁄ F Omonofluoroacetate ion

References: 1* Anonymous Press release (Thursday 2/11/2006). 1080 vital for agriculture and the environment. Royal Forest and Bird Protection Society. "it breaks down on contact with soil or water into harmless salt and vinegar, and does not persist in the environment" 2. Anonymous (August 2002) Forest and Bird Fact Sheet. POBox 631, Wellington. www.forestandbid.org.nz. "soil micro-organisms and water break 1080 down harmlessly into salt and vinegar" 3. Blackman A (2002). Inside the workings of a controversial killer. Chemistry Matters: a monthly column from the Department of Chemistry at the University of Otago. This article first appeared in the Otago Daily Times on 03 June 2002 Conclusion: The information given to the public, about the break down of 1080 into harmless products is incomplete and incorrect. This needs to be acknowledged by the agency responsible, namely Forest and Bird. The possible breakdown products when the C-C bond is broken in sunlight, over time and in solution, are sodium bicarbonate and methyl fluoride. ©MLJarman CMB 90 Owhango 3990 via Taumarunui NZ

12. What is microbial de-fluorination? References: 1 Eason CT (1993). Persistence of 1080. Surveillance 20(2):7-8. Conclusion: Certain soil bacteria and fungi are able to cleave the fluoro-carbon bond in a process called microbial de-fluorinaton.1

13. What are these "commonly occurring" micro-organisms that can de-fluorinate 1080 and how widespread and common are they? Micro-organism is a term which encompasses viruses, bacteria, algae and fungi. I have so far been able to locate the full text of the following references. There are many authors who refer to each other, but I need the full text in order to find the listing of micro-organisms involved. References: Pseudomanus-sp (unidentified), P. acidovorans, P.fluorescens

Bacteria

Fusarium oxysporum

Fungus

Penicillium purpurescens, P.restrictum

Fungus

In Australian soil/work done in laboratory Moraxella species strain B "typically found in bacterial species present in synovial tissue from human patients with various forms of arthritis"

Bacterial Pathogen (a harmful microbe)

This work was done in Australia, under laboratory conditions, GE related,

13 bacteria and 11 fungi ... .Fusarium oxysporum has by far the greatest de-fluorinating ability... This work was done on Australian soils under laboratory conditions

Fungus

King DR, Kirkpatrick WE, Wong DH & Kinnear JE (1994).Degradation of 1080 in Australian soils. Pp45-49 in Seawright AA and Eason CT (eds). Proceedings of the Science Workshop on 1080. Royal Society of NZ Miscellaneous Series 28. soils "contain a diverse assemblage of bacteria and fungi" Gregg K, Cooper CL, Schafer DJ, Sharpe H, Beard CE, Allen G and Xu J (1994). Bio/Technology 12, 1361-1365 ..growing cultures were able to detoxify fluoroacetate in the culture medium ....the construction of rumen bacteria that are able to detoxify an important natural poison supports the feasibility of using genetically modified rumen bacteria to aid animal production. Twigg LE & Socha LV (2001) Soil Biology and Biochemistry 33(2) 227-234. Soil samples (3 replicates) from central Australia were collected on seven occasions over an 8-month period, and the microorganisms capable of de-fluorinating 1080 were isolated. When grown in an inorganic medium containing 20mM 1080 as the sole Carbon source, 24 species were able to defluorinate 1080:

Conclusions: Most of the microbial work reported on has been done on Australian soils under controlled conditions. The New Zealand examples have also been done under controlled laboratory conditions. There are different motivations for the work in Australia and New Zealand. Some plants can also de-fluorinate fluoroacetate. This is still being investigated in the literature. There is no mention of the total distribution of micro-organisms in New Zealand soils.

14. What are the products of de-fluorination? Reference: Method.....The defluorinating ability of the microorganisms was determined in aqueous solution or in autoclaved soil (sterile) by determining the concentration of F in the culture broth using a Felectrode...

1. King DR, Kirkpatrick WE, Wong DH & Kinnear JE (1994).Degradation of 1080 in Australian soils. Pp45-49 in Seawright AA and Eason CT (eds). Proceedings of the Science Workshop on 1080. Royal Society of NZ Miscellaneous Series 28.

Conclusion: The cleaving of the strong C-F bond in sodium monofluoroacetate would release Fluoride ions (F-)1

15. Where are the products of de-fluorination? The Fluoride ions (F-) could be imbibed onto other hydrocarbons (organic molecules containing hydrogen and carbon - cellulose) in the upper layers of soil, attached to plant roots through mycorrhizal fungi, or in the litter layers on the soil surface. This is unlike the breaking of the C-C bond, which could produce methyl fluoride and sodium bicarbonate in water, over time and in sunlight. I would see the two processes as competing, depending on whether or not the micro-organisms are present in all New Zealand soils, and whether the conditions are right for microbial de-fluorination to take place. Either process could involve a time lag while Fluoroacetate is still in the environment. OR — rain would dissolve the Fluoroacetate, and it would be leached down into the water table, and due to the steepness of much of the terrain that is being treated (never far from a stream or water course), into streams and rivers, at levels below detection. It has not gone away. There may still be an opportunity for plants, both algae growing on rocks and water plants, to snatch Fluoride ions from the passing Fluoroacetate, before it disappears into the ocean.' This would place the products initially in the upper layers of the soil or in the litter layers on the soil surface, imbibed by bacteria onto plant cellulose, and then in the plants or algae in streams and rivers. Does this build up in the environment? What are the long term effects? What is happening to the 'biodiversity' of the micro-fauna, as they possibly go through DNA transformations in order to © MLJarman CMB 90 Owhango 3990 via Taumarunui NZ

respond to the increased availability of fluoride ions? What is happening to the invertebrates that share this environment? 2 References: 1 Eisler R (1995) Sodium Monofluoroacetate (1080): Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review. Published by the U.S. Dept of the Interior as Biological Report 27, Contaminant Hazard Reviews Report 30 February 1995 "More research on 1080 persistence in aquatic environments seems needed." 2. Aitkinson IAE, Campbell DJ, Fitzgerald JEC, Flux JEC & Meads MJ (1994). Possums and possum control; effects on lowland forest ecosystems. A literature review with specific reference to the use of 1080. DoC Report: Science For Conservation (1) 32 pp Conclusion: The products of de-fluorination could be initially in the upper layers of the soil or in the litter layers on the soil surface, imbibed by bacteria onto plant cellulose, and then in the plants or algae in streams and rivers. This needs to be researched further.

16. What are the factors controlling microbial activity? Temperature (both soil and air) 1,2, position of micro-organisms in the soil profile (near the surface or deeper)2, soil pH 3, amount of carbon available in the soil and the bait ', availability of microorganisms4, how efficient the particular type of micro-organisms are at de-fluorination 1,2.3, how long it takes them to de-fluorinate 1,2,3,5 , moisture content of soil not just rainfall 2. Research done in Australia has used baits that have been inoculated with micro-organisms to ensure that breakdown occurs 5. Is this necessary in the New Zealand situation? What would this do to the costs of producing the baits? Aitkinson et al (1994) in their literature review with specific reference to the use of 1080 6, identify research priorities. These need to be revisited. References: 1. Wong DH, Kirkpatrick WE, Kinnear JE and King DR (1991). Wildlife Research 18(5) 539-545..' In general, the de-fluorinating activity was low when 1080 was the sole carbon source, but in the presence of an alternative carbon source (eg peptone meat extract) the de-fluorinating ability of many organisms was greatly enhanced.....', `....only Fusarium oxysporum exhibited high defluorinating ability in soil, defluorinating approx equal to 72% of the available 1080 (20mM) in 15days at 270C......' 2 Kirkpatrick WE (1999). Assessment of sodium fluoroacetate (1080) in baits and its biodegradation by micro-organisms. Masters Thesis. School of Biomedical Sciences, Curtin University of Technology, Australia. `....loss of 1080 from baits buried occurred at a faster rate than from baits placed on the surface in the same time period..','.....Rainfall was recorded and temperature data was collected at each site..', 'initially 1080 loss from baits was minimal ..further loss was gradual even when it rained', 'generally baits had to be exposed to at least 50mm of rain before 1080 loss increased to 50%','..thirteen isolates showing varying degrees of 1080 degrading ability..', `the optimum temperatures for 1080 degradation were 30 degrees Celsius and fluctuating ambient temperatures of 15-28 degrees Celsius. 3 Twigg LE & Socha LV (2001) Soil Biology and Biochemistry 33(2) 227-234. ..'the fungus Fusarium oxysporum had by far the greatest defluorinating ability, and de-fluorinated about 45% of added 1080 within 12 days..... Significantly greater at pH5.6 compared to pH 6.8 ...appeared to asymptote after 2128 days...... © MLJarman, CMB 90 Owhango 3990 via Taumarunui NZ

4. King DR, Kirkpatrick WE, Wong DH & Kinnear JE (1994).Degradation of 1080 in Australian soils. Pp45-49 in Seawright AA and Eason CT (eds). Proceedings of the Science Workshop on 1080. Royal Society of NZ Miscellaneous Series 28. 5 McHroy 1981a in Eason ..........In another study, 1080 solutions prepared in distilled water and stored at room temperature for 10 years showed no significant breakdown; moreover, solutions of 1080 prepared in stagnant algal-laden water did not lose biocidal properties during 12 months 6. Aitkinson IAE, Campbell DJ, Fitzgerald JEC, Flux JEC & Meads MJ (1994). Possums and possum control; effects on lowland forest ecosystems. A literature review with specific reference to the use of 1080. DoC Report: Science For Conservation (1) 32 pp.

Conclusions: The often quoted examples of the success of microbial de-fluorination studied in controlled laboratory conditions, cannot be extrapolated into the real New Zealand outdoor environment without further URGENT, CAREFULLY DIRECTED AND COSTLY RESEARCH AND TESTING. This needs to be acknowledged to the New Zealand Public. THE USE OF 1080 SHOULD BE STOPPED UNTIL THIS IS DONE. © MLJarman CMB 90 Owhango, 3990 via Taumarunui NZ