Pigeon faeces prohibited substance screening and confirmation Dr Schalk de Kock

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2. Pigeon faeces prohibited substance screening and confirmation Dr Schalk de Kock Laboratory Director

3. The core function of this Laboratory Thoroughbred racing in Southern Africa is provided with a competent and efficient drug testing service in accordance with local and international rules. The screening and confirmation capabilities, analytical and prosecution experience can only benefit SANPO 3

4. Analytical Capabilities The list of prohibited substances being screened for within horseracing is extensive. The screening is very sensitive This requires very advanced instrumentation and techniques for the extraction, separation, detection and confirmation of these prohibited substances. There is continual investment in the most modern instrumentation to satisfy the stringent screening requirements demanded by the horse racing industry and to be cost effective 4

5. Laboratory accreditation and quality • The Laboratory has been ISO compliant since 1994 • Currently to ISO/IEC 17025Is inspected on a regular basis • This compliance include • specimen and data integrity • all operations are documented and • applied according to approved procedures • methods and screening are consistently applied at all times • screening sensitivity is constantly monitored by quality control specimens 5

6. How good is the capability to screen pigeon faeces for prohibited substances • Pigeon faeces analysis complement the screening of racehorses well. We can provide a high quality screening service due to the following: • The range of prohibited substances existing • The screening sensitivity • Much experience in pigeon faeces analysis • Extensive experience in prohibited substance prosecution • A competitive screening cost 6

7. The range of prohibited substances There is extensive experience in the screening of all the very many substances of importance to screen within pigeon racing. This comes from many years of screening in racehorses • Special attention is however of course provided for those substances considered to be most-performance-enhancing. • (please help us to consider and identify these…..) • Also those preparations and substances considered of most concern, noted to be used in pigeon racing • Confiscated preparations have been identified…. 7

8. Ten years of pigeon faeces analysis and administration study experiencesWe now enter the tenth year of experience in pigeon faeces analysis and prohibited substances prosecution. We not only have much experience of natural pigeon substance profiles (such as steroids and feed components) We also have experience from many pigeon administration trials (administration study of 16 pigeons, studying 8 important drugs)Experience also comes from positive findings from our client within South Africa, Ireland and the United Kingdom (RPRA). 8

9. The control of drug use in sport is undertaken on a worldwide scale by The World Anti-Doping Agency The WADA website indicates the substance list and requirements for humans sports. At this stage animal sports must made their own lists.It is however clear that screening/ testing for these substances is an important aspect which must be in place. Note that the drug lists for human sports and pigeon racing are not that dissimilar when considering drug classes. Any veterinarian with pharmacological experience will tell you that drugs which are active in the human are also, with a few exceptions, active in the pigeon, while some have definitely been found more active in pigeons than humans. 9

10. The control of drug use in sport is undertaken on a worldwide scale by The World Anti-Doping Agency In their World Anti-Doping Code for 2003, Article 16: Doping Control for Animals Competing in Sport the following is specified:"In any sport that includes animals in competition the International Federation for that sport shall establish and implement anti-doping rules for the animals included in that sport". From the local perspective the South African Sports Confederation and Olympic Committee (SASCOC) and the South African Institute for Drug-Free Sport are committed to abide by the code of WADA "It damages the integrity, image and value of sport, and contravenes the fundamental principle of sportmanship and fair competition". 10

11. Horseracing comprehensive or WADA (human) less comprehensive screening? • Comprehensive as animals cannot decide what to administer, humans can • Humans have rights (drink coffee and energy drinks) • Must be able to treat conditions (asthma, inhalers) • Different substances prohibited for different sports (weightlifting vs. archery) • Humans will not use life threatening or experimental substances and doses 11

12. Which substances should be screened for?  Making a pigeon win a race is the objective and this is possible with performance enhancing drugs. From the composition of confiscated preparations it is clear that the only objective is to have performance enhancing effects on the pigeon. Especially of concern are preparations and drug entities which have a longer period of drug effect than the conventional, those known to be very active in the pigeon and those which are active at very low concentration There is much knowledge in the international scientific community on which drugs are effective in the pigeon and which of these fall in this class. This comes from many years of international scientific research, clinical studies and veterinary practice. 12

13. 1. Drugs masking injury and pain: These are drugs, which prevents the pigeon from experiencing the full extent of injury in that they have painkilling properties. Experiencing injury is important as it prevents the pigeon from aggravating the injury through exercise and strain. 1.1 Narcotic Analgesics (Opiates) These are all related to the opiate morphine, drugs with very potent pain relieving properties 1.2 Analgesics Analgesics relieve pain by increasing the pain threshold. With the use of these drugs the onset of injury and injury in itself may not be perceived. 1.3 Non-steroidal anti-inflammatories (NSAID's) Non-steroidal anti-inflammatories reduce the pain of inflammation, often resulting in too much exercise considering the severity of the injury 1.4 Corticosteroids  Corticosteroids have many effects, among others the same effects as the non-steroidal anti-inflammatory drugs. They are very potent, very low doses and concentrations have effect. The beneficial effect is extended to that of well-being 13

14. 2. Drugs acting on the nervous system Some drugs acting of the nervous system can enhance performance through physical and psychological effects. 2.1 StimulantsStimulants enhance physical and psychological performance. Heart rate and blood pressure can increase and the risk of cerebral haemorrhage (stroke) and cardiac arrest is increased. Note that caffeine is contained in coffee and tea and theobromine in cacao products. 14

15. 3. Drugs improving athletic performance These drugs improve the athletic ability of the pigeon and can be endogenous (naturally occurring in the body) or non-endogenous substances. Some of the drugs have in themselves an effect while others induce physical changes to the body, or induce changes in the level of endogenous compounds. 3.1 Bronchodilators These have the effect of dilating (widening, relieving constriction) the bronchial passages. 3.2 Anabolic steroids These compounds have anabolic effects on the body. These synthetic compounds can be identical in structure to naturally occurring anabolic steroids (e.g. testosterone) or related in structure. 3.3 Synthetic Hormones This drug class refers mainly to synthetic protein hormones which are similar in structure to endogenous hormones. The most significant hormones in this class include the synthetic human analogues of insulin (regulates carbohydrate metabolism), growth hormone (has anabolic effects and promote growth) and erythropoietin (stimulates and increase in mature red blood cells). 15

16. So we simply screen for those substances which are • administered? • Not as simple as that. Most substances are metabolised (modified) • Pigeon has fast excretion and significant metabolism • Some substances ONLY detected and can only be prosecuted as metabolites • We must screen for the RIGHT metabolites • Those are not always the same as found in the human 16

17. Why metabolites / metabolism? • Body sees substance entering the body as foreign (not normal food..) , there has to be the fear could be toxic • Body tries to excreted as fast as possible, modifies drug molecule • Modifies to be more easily excreted (more water soluble) • Oxidise to different molecule (phase I metabolite) • Links to molecule to enhance excretion (phase II metabolite) • Modifies to detoxify the molecule (now a new inactive molecule) 17

18. Study of metabolites can be important during positive findings • Can provide indication of: • Preparation and route of administration (oral, injected … ) • When likely administered • The dose administered 18

19. The screening sensitivity • The matrix of pigeon faeces is not easy for the detection of low concentration of many of the prohibited substances of importance to pigeon racing. • This laboratory is well experienced in the analysis of horse urine (difficult matrix as has high background and viscosity) and then also the analysis of blood of low sample volume. 19

20. The screening sensitivity • Modern drugs are very potent, very low doses are required… • It is know and we have also shown that most of the many hundred substances of importance we can routinely screen at low part per billion sensitivity in pigeon faeces • 1 part per billion • 1 ppb is 1 part prohibited substance in 1 000 000000 parts……. • 1 second in 32 years……… • This is not far below the concentration which reflects that the substance has an affect on the pigeon… and where there should be a positive finding 20

21. The importance of a Veterinary Committee Prohibited substance policies and decisions! • Which substance classes / types prohibited? • Which substances within these prohibited? • Which substances not prohibited? • Natural substances require population and administration studies • Data must be investigated and decision must be made at which concentration to prosecute • To re-establish and maintain prohibited substance lists • Must be aware exactly what the laboratory screen for, as a suggestion 21

22. Keeping the prohibited substances classes up to date The association between national pigeon racing and the Laboratory is important in this regard. The National Horseracing Authority have close to 20 years of experience in detecting new drugs effectively and cost effectively, researching new drugs, keeping with international trends and scientific literature and they belong to international horseracing and forensic associations. Referring to these animal sport testing laboratories assure that screening is highly inclusive of important drugs while drugs which are not important (not performance enhancing) are not tested for at an unnecessary cost (screening must be sensible) 22

23. Comparatively low screening cost • With pigeon faeces analysis quite similar to the screening we provide for the racing industry and other equine sports, analysis slots in our existing, comprehensive methodology: • Very cost effectively slots into our routine screening and employ the same cost effective techniques (with consumable attractively pricing from bulk buying power) • Very advanced instrumentation for high throughput analysis • Automation and significantly automated data processing The screening is “horseracing comprehensive” , not human WADA which is limited and cheaper 23

24. Comparatively low screening costSpecimens which require positive analysisSpecimen which are found to contain prohibited substances are re-analysed several times, ultimately as an analysis separate from all other specimens • In direct comparison with blank specimensIn direct comparison with a certified reference standard of the prohibited substance. • Such analysis and the full analysis reports which accompany the result has an analysis pricing of about R4000 24

25. Sample securityFrom many years of horseracing experience we understand, respect the importance of sample security and respect the security mechanism required. • Specimens are resealed and are stored sealed • Several re-analyses are conducted prior to positive reportingThe SANPO supplied kit security has not been successfully challenged • The kits are not expensive 25

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28. Specimen collection We must have enough faeces to conduct full screening and allow for re-analysis and the confirmation of positives Must be enough in Reference B specimen also! Basket evening (pre race) specimen collecting and testing is really the only effective means of screening The metabolism and faeces excretion of the pigeon is fast 28

29. Specimen collection There is always the option to collect many specimens on basket evening with proper security and labelling (the specimen number only, identification must only be possible on a separately kept document) Always keep pigeon faeces in the freezer, never in fridge only Can keep for long but must not freeze and defrost Overnight post office courier is fast and reliable for this purpose 29

30. Receipt of specimens at our laboratory Upon specimen receipt at the laboratory a record will be made of the condition of the specimen(s), the intactness of seals and the seal numbers. Any irregularities are immediately reported to the client as to inform that it could affect the prosecution of a positive finding Routine prohibited substance screening at the laboratory only involves the analysis of the Original (A) specimen. The Reference (B) specimen is also received and kept safely 30

31. Specimen analysis and reporting • Specimen is opened, take small amount for analysis, resealed • Hydrolysed (phase II metabolism considered) • Extract only substances of interest for screening • Make sure no losses of prohibited substances of concern • Clean up for best screening sensitivity • Longer instrument analysis for best sensitivity and detection • Advanced and sensitive instrumentation • Cost effective and fast data analysis • Reporting of negative results typically within 7 working days (Positive finding confirmation takes longer) 31

32. Positive findings • Negative result is quicker in reporting • Positive result requires several retests • Positive result introduce option of Reference (B) specimen analysis • Must be requested within a specified period of time • Must be at our laboratory • Specimen is not kept for long if negative, kept until case is closed for positives 32

33. Screening into the future • Pricing?The more analyses, the bigger the analysis batches, the lower the cost can be… • Specimen numbers?More specimens, more money more drug trials and other “natural substance level” investigation work possible • Blood screeningFoot prick for blood drops can be a suitable screen with suitable sensitivity? • Feather analysisProhibited substances are incorporated into the feather (similarly like into human hair) 33

34. Where prohibited substances originateSome of the preparations are being made available by veterinarians. The administration or supply of any drugs by a veterinarian without a clinical justification, such as in the case of deliberate doping, will definitely be reported to the Veterinary Council. The administration of registered drugs to any animal without a written veterinary prescription by a non-veterinarian is illegal. During abuse prosecution cases where this practice is clear the person responsible for the abuse could face other charges also. Such practices will in some cases be reported to the local Department of Health who may wish to issue fines or prosecute legally in a separate case(s). The possession of an unregistered drug preparation and the purchase of such preparations are prosecutable by customs and excise. 34

35. Concern over substances being are prohibited Contact your veterinarian to assist with the identification of prohibited substances and also refer to the SANPO list of prohibited preparations. Based on the classes of drugs specified in the SANPO rules it is easy for a veterinarian to know which drugs and preparations are not allowed. For most of the prohibited drugs, should these be prescribed by your veterinarian your pigeon is probably sick or injured and should not be racing. It may be a good practice to check the supplements and other preparations you routinely administer to your pigeons with your veterinarian in order to ensure that these are not prohibited. 35

36. Concern over substances being are prohibited In theory almost all prohibited substances in pigeon racing are only available as a prescription from a veterinarian or doctor or available from pharmacies. Only a few preparations such as caffeine (contained in some teas and coffee) and headache tablets (aspirin and paracetamol) are available via on the shelve retail 36

37. Are prohibited substances being used? There are those boasting that “they can’t catch us”, “they don’t test for this” and “they cannot detect this”, irrespective of how much drug testing is undertaken. Many are using natural products and supplements which are not prohibited substances, but are very convinced themselves and convincing to others that they are. Many are under the impression that the preparations which they are using are making a difference. This is often not true as clinical and a real scientific evidence show that the preparations involved are often not physiologically effective in this regard. The only effect of these preparations and resulting rumours are to upset others and create unnecessary excitement. 37

38. Substances which are not prohibited The views from forensic screening laboratories and clinicians in the field are in consensus in stating that the important substances are being tested for while those which are not important are being ignored. Substances beneficial to the health of the pigeon which are not prohibited Note that almost all medications and preparations for pigeons which are important to the pigeon’s health and welfare are not prohibited in racing Anti-bacterials(e.g. for the treatment of coccicidosis, trichomoiasis – crop cancer, ornithosis,mycoplasmociscandida, salmonella etc.)……………. 38

39. Substances beneficial to the health of the pigeon which are not prohibited …… anti-viral preparations, anti-oxidants, anti-parasitics, amino acids, blood formation agents, detoxifiers, dewormers, energy boosters, herbs, insecticides, immune activators/boosters, minerals trace elements, vitamins, vaccinations. Supplements of natural occurring intermediates (e.gcreatinine, methylsulphonylmethane – MSM, L-arginine, L-carnitine, nicotinamide, biotin, methionine, niacin, dimethylsulfoxide - DMSO, glutamine, taurine, choline, folate, riboflavin, thiamine etc.). These are (or possibly could be) beneficial to the health of your pigeon at the appropriate times and doses, can of course be administered to the race pigeon, and are not prohibited. 39

40. Substances beneficial to the health of the pigeon which is not prohibited Homeopathic preparations These are not prohibited for pigeon racing and real proof that these have real significant effects on the performance of the pigeon, horse, human or other animals is arguable. 40