Research Papers
Council Inc

     by Dr Diane Ryan, Chairperson

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Submitted to the Australian Harness Racing Council Inc
28 March 2001

FOR GENERAL RELEASE       Release Date:  10 April 2001

1.  Terms of Reference
2.  TC02 Review Sub-Committee Participants & Others
3.  Report 
4.  Supporting Papers & Documentation
Other Select Published Papers
on Feeding, Nutrition etc, Dealing With Aspects Relating To Equine Welfare Given The Results Of Standardbred Diet Survey



That a Sub-Committee be created, to be chaired by Dr Diane Ryan to conduct an evaluation of the existing material available to Council on TC02, and to have the power to co-opt the necessary expert advice and opinions in order to arrive at a recommendation to the AHRC of the:

1.  Level of laboratory uncertainty factor in the testing process
2.  Integrity of the testing process
3.  The mean TC02 level of the harness racing population
4.  The mean TC02 level of untreated harness racing population
5.  Has the mean level increased and reasons why?

Report to the Executive with a Summary and Recommendations within 12 weeks, then the Executive to consider, prior to referral to Council Members.


Sub Committee Participants

  • Dr Diane Ryan (Chair) BVSc, PhD (Chair)
    Deputy Chairperson, Harness Racing New South Wales
  • Dr Derek Major, BVSc, MACVSc.
    President, Australian Equine Veterinary Association
  • Dean Cooper, LL.B(Hons) (University of Tasmania), Barrister & Solicitor
    Legal Counsel, Australian Harness Racing Council
    Partner, Phillips Taglieri, Hobart

Racing Forensic Laboratories

  • Dr David Auer, Director (A/g), BVSc, BSc, DipEd, PhD
    Racing Integrity Services, Racing Science Centre, Queensland
  • Dr John Vine, Director, BSc, MSc, PhD, FRACI, FAORC
    Racing Analytical Services, Victoria
  • Dr Rob Rahaley, Director, BSc, MSc, PhD, Dip.ACVT, MACVSc
    IDEXX Veterinary Pathology Service, Adelaide

Independent Consulting Biometricians

  • Biometrical Consulting Service
    The University of Sydney
    • Associate Professor Mick O’Neill
    • Ms Kathryn Bartimote
    • Dr Peter Thomson

Australian Harness Racing Council, Corporate Services

  • Rod Pollock, MBA(Melb), FCPA, JP (Secretary)
    Chief Executive, AHRC
  • John Potter, Steward – Industry Liaison
    Charles Skillen, Stewards Panel
    Harness Racing Victoria, Melbourne

Standardbred Diet Survey

  • John Kohnke BVSc RDA
    John Kohnke Consultancy, Equine Feed
  • Dennis Roberts B.Com (Melb), MBA (MGSM)


1.     Professor Reuben Rose, Faculty of Veterinary Science, University of Sydney withdrew from Sub-Committee on 19th March 2001.

2.     The Final Report to the AHRC Executive has been prepared by the Review Sub-Committee comprising Dr Diane Ryan, Dr Derek Major, Dean Cooper and Rod Pollock (Secretary). Neither Professor Rose nor any participant from the Racing Forensic Laboratories has been involved in its preparation.

Racing Laboratories

The Racing Forensic Laboratories information utilised in the Plasma samples analysis were from:

  1. Racing Science Centre (RSC) Queensland
  2. Racing Chemistry Laboratory (RCL) Western Australia
  3. Australian Racing Forensic Laboratory (ARFL) New South Wales
  4. Racing Analytical Services Limited (RASL) Victoria and utilised by South Australia and Tasmania.
  5. IDEXX Veterinary Pathology Service.


Sample Data was provided by the Laboratory or the State Controlling Body.



The intention is that all racing laboratories involved in the testing programme:

  1. Are all accredited by NATA or equivalent,
  2. Apply a common Standard
  3. Utilise one Process Methodology.


The principle of measurement of TC02 must be identical throughout Australia.


  1. There must be the same threshold and the same allowance for the measurement of uncertainty.
  2. Similar data must be collected by each State Controlling Body.
  3. A common policy of disclosure of tested results should be developed and communicated.


a.    Laboratory procedures and methodologies including recognising the requirements of the International Laboratory Accreditation Co-operation should be regularly reviewed as part of the NATA Accreditation.  

b.    All testing laboratories must standardise their methodology and processes. Certified referee material must be in place.

c.    It was recommended that five (5) samples should be taken to effectively complete the process of analysis. One primary sample is required as the screening sample with the other samples required for any subsequent analysis.

d.    It was suggested that a smaller tube (5ml container) with a narrow bore/gauge needle be utilised for collection and animal welfare reasons.

e.    All laboratory processes including instrument calibration and equipment testing should be standardised in all States.

It is noted that the Beckman Synchron EL-ISE has been in use in all 4 Australian Racing Laboratories since 1992. Further that methods have been standardised and that the same batches of calibrators, controls and standards are currently in use.

f.    The AHRC/State Controlling Bodies should develop a specific uniform laboratory contractual agreement to undertake and operate this testing program nationally.

g.    Significant laboratory networking, peer review and inter-laboratory collaboration beyond the current formal referee sample system should be encouraged and adopted in a formal system.

h.    Consideration of the logistics of transportation between Western Australia and other States is an important consideration given the airfreight time involved. A range of benchmark requirements should be mandated to operate across the "collection to analysis" cycle.

i.    That all laboratories nationally develop and update a standardised reporting system. There should be not be any truncation or artificial factoring of any results from the testing program as has occurred in Victoria, Western Australia and South Australia in the recent past. Whilst acknowledging that this approach has been of benefit for the trainer through this conservative approach of under-reporting; accuracy in the raw results is essential.

It is recommended that the Racing Laboratories exercise no independent judgement on the testing results.

j.    That any "observable" impacting changes to any part of the laboratory’s processing operations or methodologies be reported to the AHRC/State Controlling Body within one (1) month with an impact statement detailing the occurrence, its effect on the testing program and any necessary action implemented or planned. This will enable the Regulatory Body to consider its approach for industry participants.

k.    That each State Controlling Body record and compile standardised information on its testing program utilising the national computer system. Reporting information on the testing programs results to our industry should occur with a defined lag.

l.    In the International Agreement for the Federation of Horseracing Authorities in Clause 18 it states that their laboratories should:        

  1. be accredited according to ISO/IEC 17025, General requirements for the competence of testing and calibration laboratories, and to the supplementary document ILAC-G7, Accreditation requirements and operating criteria for horseracing laboratories
  2. conform with the Guide for establishing the presence of prohibited substances (Part B of ILAC-G7)
  3. meet the Performance specification of the International Federation of Horseracing Authorities (Part C of ILAC-G7)
  4. take part in interlaboratory comparisons (clause 5.9(b) of ISO/IEC 17025:1999).

m.    Additionally the International Agreement in Clause 19 suggests that "with the objective of preventing infringements, the administrative Body may:-

  1. make available detection times
  2. give forewarning of new or modified tests
  3. provide an analytical service to establish whether a sample from a horse entered to race contains prohibited substances the trainer specifies

This is agreed with subject to the addition of relevant time frames for the first two instances.


a.  Throughout many submissions and other industry documentation much has been made of comparisons with both the Australian Thoroughbred Industry (36mmol/L) and the Accord of the International Federation of Horseracing Authorities (37mmol/L), the International Agreement.

b.    In firstly considering the International Agreement it should be noted that it is designed for the guidance of the Racing and Breeding Authorities internationally so that … in their own Rules of Racing and Breeding, they may aspire to the absolute protection of the integrity of horseracing. It should be noted that the Australian Thoroughbred Industry is not a signatory to the Agreement in respect of the level of 37mmol/L. It has varied the agreement for the area defined as Section 5, Sanctions and the threshold level to 36mmol/L. This is a significant statement in this international forum.

c.    Reference was made to the United States and their policies. Enquiries were made to the United States Trotting Association, Professor Richard Sams Laboratory Director of the Analytical Toxicology Laboratory at The Ohio State University provided a response based on the Ohio jurisdiction. The response is attached for information. It is noted that there are many similar areas to Australia. They utilise a level of 37mmol/L. However the United States of America and Canada are also not signatories to the International Agreement referred to above.

d.    It is considered that these factors should be noted in Council’s evaluation on the Australian TC02 threshold level. In addition, reference should be made to the article by Dr David Auer, Director of the Racing Science Centre, Queensland. It is entitled "Facts pertaining to establishing a threshold concentration for the total plasma carbon dioxide concentration of Australian harness horses.


a.    The initial work done in developing the threshold of 35mmol/l was undertaken in the early 1990’s. The results of experiments conducted at Harold Park and then subsequently for 95 Standardbred horses in the Bankstown, Fairfield and Menangle areas from 20 different racehorses were analysed on three machines the Beckman EL-ISE Autoanalyzer, Ciba-Corning Fast 4 Autoanalyzer and Radiometer ABL 300 blood gas machine in the Equine Exercise and Drug Evaluation Laboratory at the Department of Veterinary Clinical Sciences, University of Sydney. The result was a very narrow distribution between 25.4mmol/L to 32.7mmol/L. It was important to note that in this initial work the tc02 values as measured by the Radiometer blood gas analyser were on average 1.4mmol lower than the TC02 concentration measured on a Beckman-Elise Autoanalyser, which is the machine currently used by the Racing Laboratories. The results of the Population Study were:

Machine n HC03 (mmol/L) TC02 (mmol/L)
Blood gas machine 95 29.3 ± 1.4 30.8 ± 1.4
Beckman 89 - - 32.2 ± 1.3
Ciba-Corning 91 - - 30.5 ± 1.6

b.    Council adopted a threshold of 35mmol/L with an actionable level of 36.2mmol/L in all States with the exception of Western Australia with a level of 36.4mmol/L. The Beckman is in use as the key piece of equipment in the Racing Laboratories now. (Refer Dr David Auer’s Articles).

c.    Advances in technology with its procedures and processes over time have meant that consideration of the threshold is also appropriate. The information contained in the Biometrician’s Report and the Standardbred Diet Survey reveal changes to a number of areas not evident in the early 1990’s. The significant feeding patterns utilising different combinations indicates normal usage of electrolytes, citrates etc. The knowledge of products and their combination cannot be estimated or known in an industry of professionals, semi-professionals and hobbyists however the Diet Survey is indicative.

d.    Although Council consensus on a threshold level of 35mmol/L was achieved there was considerable discussion prior to agreement. The principal reason for setting the threshold level of 35mmol/L was to counter the practice of "milkshaking" and masking/disguising other prohibited substances. There is evidence to suggest that in the current environment innocent action may result in exceeding the threshold level.

e.    The Biometrician has reported that assuming that the data basically represents a population of untreated horses then between 1.8% (the Queensland estimate) and 2.8% (the South Australian estimate) of samples will naturally occur above 35mmol/L. That is if this is considered to be a valid assumption. That is, if 35mmol/L is the threshold limit then between 18 and 28 in every 1000 samples will be detected as having excessive TC02 levels. They may or may not exceed the actionable level. Their conclusion is that this would appear too high. The statistics are not normally distributed. Refer Dr David Auer’s Articles on the statistical concepts relative to Type 1 and Type 2 errors and their implications.


The Australian Laboratories have an explicit allowance for the measurement of uncertainty. Racing Laboratories are participating in the preparation and implementation of ISO 17025, which requires laboratories to estimate their uncertainty of measurement.

The following should be contemplated:

  1. The adoption of NATA accreditation and the new Standard (ISO 17025) will reinforce and regularly review this area.
  2. The possibility of a lowering of the measurement of uncertainty factor because of ongoing improvements in testing methodologies.
  3. The measurement of uncertainty factor should be reviewed annually or bi-annually.

As a direct result of the change from CASCO to ASE the Racing Forensic Laboratories are engaged in work on this area. The Review Sub-Committee considers it important that the testing program nationally be considered as one laboratory with different sites standardised and consistent. This calculation is to cover anything that happens in a laboratory so as not to cause variation in testing. The pursuit of accreditation both nationally and internationally further aids this process. This will also further develop the standards in the control material themselves. The standard is what is used to determine the measurement of uncertainty. The use of the control material is important for establishing the rules of rejection in machine calibration for testing. The laboratories already do a significant amount of interlaboratory collaboration. The highest standard possible together with quality benchmark controls/management is essential in this area. It is considered that one laboratory uncertainty of measurement should apply throughout Australia.


a.    Council was first made aware of changes in these Standards at its Special General Meeting in May 2000 in Melbourne. Reference should be made to Dr John Vine’s Paper on Measurement of TC02 in Australia "Pages 2 to 4".

b.    The CASCO (CASCO-NERL Diagnostics, Maine) Standard Change to ASE (Australian Scientific Enterprises Pty Ltd) is now well documented and communicated. The laboratories have advised on the technical steps taken to maintain the integrity and accuracy of the testing process after identification and during the changeover transition period. The laboratories adopted methods of truncating the results and factoring data downwards to reduce the reported values. South Australia adopted an even more conservative approach at the time. This action favoured the industry participant in the past.

c.    Discussion with the Racing Laboratories has ensured that in their opinion the necessitated change has resulted in a more accurate/robust calculation of the TC02 measurement. This is a direct result because the instrument is now calibrated between 27mmol/L and 39mmol/L. The statistics as provided by the Biometrical Consulting Service show a range of information, which will be summarised later. However, an important indicator is presented by the Table entitled Means of TC02 values on a State-by-State basis. The part of the Table refers to the immediate period between the last CASCO period and the next two ASE periods. The data is:

Means of TC02 values on a State-by-State basis.
(Counts of TC02 Values)









    1998 (CASCO)







    1999 (CASCO)







    1998 (ASE)







    1999 (ASE)







    2000 (ASE)







    By observation, it would appear that the mean has risen in this transition period. The frequency is included to dimension the sampling undertaken. Consider in association with (b) immediately above.

d.    Where any change occurs the Racing Laboratories must communicate such change to the relevant State Controlling Body and AHRC.


a.    The Policy on collection of samples for all States must be uniform, standardised and published. The policy should clearly define all aspects of the collection, handling, transportation and hand-over of samples to the Racing Laboratories. Timeframes must be mandated to operate against.

b.    It is considered important that further work be conducted and policy implemented on the following areas:

  1. A policy on selecting horses to be tested that is random and scientifically based without prior communication for integrity reasons.
  2. A comprehensive Statewide/National testing program designed to be cost effective and scientifically based to be implemented with statistical design and comprehensive coverage.
  3. Consider whether emphasis on any categories is required i.e. all winners etc.
  4. Further determined design and policy for pre and post race testing as part of the program.
  5. Further evaluate and determine a security policy that is relevant to the testing program in its entirety.

c.    Establish and monitor compliance of the testing program and its data through the statistical data collected. There is a lack of comprehensive consistency in the current testing programs and data collected.

d.    A specific education process be developed for distance learning and internet/seminar delivery for harness racing industry participants.

e.    Council has accepted that technological and other environmental changes between the trainer, the racing scene and the standardbred are adequately catered for through improvements undertaken by the Racing Forensic Laboratories over time. It has not previously reviewed this important area. Future review needs to occur periodically. They need to be open and further gain the best scientific evidence available of matters, which are researched adequately to reduce any impacting variables influencing the testing program. In this way external impacting variables can be minimised.

f.    Identify, design and fund a number of research studies on areas of investigation in conjunction with appropriate scientific resources in conjunction with the Australian Racing Forensic Laboratories.

The effects for example of the longer continued usage of low dose drug administration and whether it has a cumulative effect. The administration of alkalising salts etc. maybe another as is the legitimate use of therapeutic drugs.

David Auer’s Paper is important here as it suggests that there is an important need to balance the needs of trainers, the welfare and integrity of the racing industry together with the interests of the horse.


  • The Report of the Review Sub-Committee to the Australian Harness Racing Council Members was provided in a Video Conference held on Monday 9th April 2001.

  • These papers are not for general release as they will be discussed in Council’s Meeting on Monday 30th April 2001. Please note that the confidentiality aspects on the Written Submissions, Scientific References and Other Presentations have been maintained. The summary report delineates the issues and areas raised by the submissions etc. tendered.

  • The various papers are identified and listed:   

    • Terms of Reference
    • TC02 Review Sub-Committee Participants and Others
    • AHRC Rules on Prohibited Substances
    • Report
    • Facts pertaining to establishing a threshold concentration for the total plasma carbon dioxide concentration of Australian Harness Horses - Dr D. Auer.
    • Measurement of TC02 in Australia – Dr J. Vine
    • Report on an Investigation into Pre-Race TC02 Levels of Australian Harness Racing Horses 1995-2000 by Biometrical Consulting Services, The University of Sydney, New South Wales.
    • Measurement of plasma total carbon dioxide concentrations in stabled Thoroughbred racehorses in Australia using the ASE Linearity Set – Dr D Auer.
    • The total plasma carbon dioxide threshold – think about it – Dr D. Auer.
    • Report on Written Submissions, Scientific References and other Presentations.
    • Report on Standardbred Diet Survey and Article – Alkalising Agents – Food for Thought by Dr D Auer.
    • International Agreement on Breeding and Racing and Appendixes – International Federation of Horseracing Authorities.
    • Referral Response from USTA provided by Analytical Toxicology Laboratories, The Ohio State University.
    • ILAC – G7: 1996
      Accreditation Requirements and Operating Criteria for Horseracing Laboratories.
    • TC02 – Questions & Answers – Australian Harness Racing Council


  • Kentucky Equine Research Inc
    W.A. Stutz and others
    Oklahoma State University.

Paper: Effects of Dietary Cation-Anion Balance on Blood Parameters in Exercising Horses.

  • Equine Nutrition and Physiology
    Society, 12th Biennial Meeting
    1991, Canada, L.A. Baker and others

Paper: Effect of Dietary Cation-Anion Balance on Acid-Base status in Horses.

  • Oklahoma State University,
    J.C. Popplewell and others

Paper: Effect of Dietary Cation-anion Balance on Acid-Base Balance and Blood Parameters in Anaerobically exercised Horses.

  • Oklahoma State University,
    L.A. Baker and others

Paper: The comparison of two forms of Sodium and Potassium, and Chloride versus sulfur in the Dietary Cation-anion difference equation: Effects on Acid-Base status in Sedentary Horses.

  • University of Illinois,
    Kimberley F Kauffman and others

Paper: Effects of Diet on Plasma TC02 in Horses.

Release Date: 10 April 2001

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