Calicivirus Studies Laboratory

From:   Alvin W. Smith, DVM, PhD
	Professor, Oregon State University
	College of Veterinary Medicine
	Laboratory for Calicivirus Studies
	Corvallis, Oregon  97331
	Fax 541 737 2730

To:  	National Registration Authority    
    	(For Agriculture and Veterinary Chemicals)  
     	Attention: Joan Ashton, BS, BVSc           
     	First Floor Industry House               
     	10 National Circuit, Barton ACT           
     	Fax 616 272 4753

An open letter:     ADDRESSING THE PROPOSED APPROVAL AND OFFICIAL
                    RELEASE OF RABBIT HEMORRHAGIC DISEASE (RCD) 
		    AS A BIOLOGICAL AGENT

With Copies to:

The Hon. John Howard                            Rt. Hon J. Bolger
Prime Minister                                     Prime Minister
Fax 2 251 5454                                       Fax 473 7045

The Hon. John Anderson                    Hon. Dr. Lockwood Smith
Minister for Primary Industries & Energy     Min. Of Agriculture 
Fax 6 273 4120                                       Fax 471 2918

The Hon. Dr. Michael Wooldridge                Hon. Peter Topsell
Minister for Health       Speaker of the House of Representatives
Fax 39 563 4993                 (For the attention of Parliament)
                                                     Fax 472 2055

The Hon. Michael Lee                          Hon. Winston Peters
Shadow Minister for Health                Leader N.Z. First Party
Fax 4 334 3422                                       Fax 472 9723

Senator Cheryl Kernot                                Helen Hughes
Democrats Leader          Parliamentary Comm. for the Environment
Fax 73 844 3671                                      Fax 471 0331

Senator Bob Collins                              Director General
Shadow Minister for Primary Industries         Ministry of Health
Fax 088 981 3040

EXECUTIVE SUMMARY

To be approved as an "Agent Organism" RHD/RCD must first be proven to be safe and host specific for the feral rabbit Oryctolagus cuniculus. However, there is a virtual certainly that the RHD/RCD agent is not single host species specific. The evidence for this statement needs to be carefully examined using all the relevant information available, and the validity of this evidence must be considered from a sound scientific point of view without special interest bias.

RHD introduction and spread presents a potentially dangerous disease situation for the livestock, wildlife, and people of Australia and one that they have needlessly been exposed to. RHD has not met the proofs of host specificity and therefore cannot be declared an "Agent Organism" in Australia. The most responsible position that the New Zealand government could take at this time would be to block the import and release of RHD and disassociate themselves from the twisted data being reported by Australian and New Zealand agencies promoting RHD release.

The following information makes up a substantial body of evidence that weighs heavily toward RHD/RCD lacking host specificity. Against these data, claims of "no evidence" or a negative research result are used to support the declarations of RHD host specificity.

ORIGINS OF RHD: The known information involving the origins of RHD is sufficiently compelling that one must assume the agent originated from a non-rabbit source and for that reason one must also assume that RHD is not species specific. This cannot be overcome without proof that RHD originated entirely from rabbits. Host specificity for the European rabbit has to be proven before RHD can be approved as a biological agent. RHD cannot gain such approval unless the matter of origin is resolved in a manner that would eliminate all non O. cuniculus species as the source, or specifically prove (not imply, but prove) that the European rabbit was the sole source of the agent.

RHD/RCD VIRUS ACQUISITION: On the basis of world wide opinion, the Australian and New Zealand groups promoting the release of RHD/RCD for controlling feral rabbits are very much isolated. These pro-RHD groups may rightfully claim that U.S. and European scientists who criticize them fail to fully understand their rabbit problems. While this may be true, it becomes increasingly apparent that the groups in Australia and New Zealand promoting RHD release fail to comprehend the nature of this agent which causes deadly disease, nor do they consider the potential for disaster that could come from the RHD/RCD release (escape) and it's subsequent spread. Confirmation for this is seen in the recent decision that an environmental impact statement would not be required.

PURE CULTURES OF THE RHD AGENT: The RHD/RCD virus cannot be propagated in the laboratory and for that reason all sources of virus for vaccines, diagnostic reagents and infectivity work are from ground-up diseased rabbit livers. As a result there is not proof that RCD is a disease caused by a calicivirus. Vaccines of crude liver proportions have received numerous anecdotal reports of harming and killing immunized rabbits. There is no vaccine for humans or other animals should they be needed.

VIRUS PREPARATION: These remain crude preparation in the sense that the source material and procedures are not sufficient to separate different classes of viruses or diffeent viruses within the caliciviridae. Test reagents thus produced become suspect.

COMPETITIVE ELISA: There are a number of limitations with this test, particularly when improperly used or when the results are misinterpreted. This has resulted in a web of disinformation regarding antibody tests and possible infections. Simply put, the competitive ELISA test was manipulated to allow questionable and seemingly positive test results to be reported as negative.

ANIMAL INFECTIVITY STUDIES: All of the test species (29) except Kiwis were tested with the competitive ELISA no later than 14 days post RHD injection, and all but perhaps mice were said to be negative. Even 7 of 7 RHD infected rabbits that survived infection tested negative at 14 days.(1) It appears the test protocol using this test at 14 days was designed inadvertently or otherwise such that negative test results could be reported.

Bats appear to have sero converted.(6) Using the competitive ELISA, their antibody levels at 14 days were as high as infected rabbits at 14 days, but the bats were reported as negative. Using the competitive ELISA one Kiwi developed higher antibody levels at day 35 than two experimentally infected rabbits at day 35 (70% for the rabbit and 78% for the Kiwi)(1,6) yet the Kiwi was reported to have not been infected and the antibodies were said to have not been the result of infection, whereas the rabbits with lower antibody levels at day 35 were known to have been infected.

The positive Kiwi tests were passed off as bad laboratory procedures, that is, failing to properly adjust the virus concentration for Kiwi infectivity studies, inside the Wallaceville biocontainment unit. A sentinel rabbit dying of RHD in an infected bird pen even though the rabbit had no direct contact exposure (reference the Wardang Island direct contact rabbit experiments and their poor success) was explained away as bad laboratory procedures inside the maximum biocontainment facilities at AAHL. Was the experiment repeated or were complete answers given as to exactly why and precisely how the sentinel rabbit became infected?

RHD/RCD ANTIBODIES IN NON-EUROPEAN RABBIT SPECIES: The report of a hare in China dying of RHD and then having it's tissues used to reproduce RHD in rabbits appears to have been purposefully overlooked in a frequently cited reference on RHD host range.(2)

Recently reported accounts of RHD antibodies in Chinese, German, and Polish hares, foxes, and dogs as well as dogs shedding RHD virus are ignored or dismissed.(5,7,9,10)

POSSIBLE HUMAN INFECTION: Extensive meaningful serologic surveys (and tests other then CSIRO's competitive ELISA) appear not to have been done on humans that have had direct and meaningful RHD exposure. Remember, the existing information on antibodies in an exposed Mexican worker indicates human susceptibility to RHD infection. Have in-depth and properly designed experiments with primates been completed to help answer the human susceptibility question? New Zealand still has the option of not exposing their people to this disease. Forty one other nations have lost that option. Four of the five calicivirus groups are known to cause disease in humans. There are no calicivirus vaccines for humans and if the ground-up diseased liver preparations used in rabbits are unsafe and unacceptable for human use, how will the authorities address possible infections in he other species including humans? Why in the RHD infected areas of Australia has there not been any publicized widely distributed laboratory tests, serologies, and specific data regarding RHD diagnosis on Australian animals other than rabbits? Such data could be a very useful tool for the agencies promoting RHD release if testing had been carried out in depth and if all the results were reliably negative and if reliable tests are carried out not using the "fixed" competitive ELISA test. Such findings would help support their contention that RHD is species specific. Are there any RHD positive tests occurring among non-rabbit species?

WARDANG ISLAND: The virus escaped containment on Wardang Island not once, but twice. If the first escape failed to point out the failing of the system, then that makes the second escape even more condemning of the procedures or the motives on Wardang Island.

Finally, there have been repeated confessions of bad laboratory procedures which have been used to explain away results not supporting a position of officially introducing and releasing RHD in Australia and New Zealand. Laboratory tests are adjusted in ways which excludes results that would complicate a pro-RHD introduction agenda. Twenty nine experimental animal species were tested at time intervals (up to 14 days) that would virtually ensure the desired negative test results. Compelling evidence of host non-specificity for RHD such as antibodies in mice, Kiwis, bats, dog, foxes, hares and a human are rejected as meaningless and a documented and reported case of the disease in a hare is overlooked. The only easily recognized success coming from an otherwise ill conceived Wardang Island field project was to provide a mechanism whereby the RHD virus could be removed from the high level biocontainment at AAHL and loosed into feral rabbits in Australia without first passing through all the regulatory and societal approval processes. The Australian and New Zealand governments are considering approval of the agent causing rabbit hemorrhagic disease (RHD) as a biological agent for use in controlling the European feral rabbits in their two countries. To be approved as an "Agent Organism" RHD must first be PROVEN to be safe and host specific, that is, it must infect only one species, the European feral rabbit (Oryctolagus cuniculus). Because this requirement is pivotal, spokespersons for the Australia New Zealand Rabbit Calicivirus Disease Program (ANZRCDP) and RBAG (Rabbit Biocontrol Advisory Group) (a) and other similarly motivated agencies have used a massive public disinformation blitz to create the perception that RHD is safe and host specific, and they have attempted to discredit any opposition that does not support their predetermined objective of releasing RHD to kill feral rabbits. Officially releasing RHD in Australia now may be of less consequence to the Australian ecosystems because the virus has already been spread to all states and it cannot be controlled. However, deliberately blanketing the continent with the virus (twenty infected rabbits released at each of 280 sites across Australia) is the epidemiologic equivalent of a wild forest fire compared to back burning through a forest. The risk for non-target species becomes greatly complicated because of the increased intensity of exposure to overall levels of RHD. In addition, failure to officially declare RHD as a "biological agent" could still be of tremendous importance to certain individuals and agencies if liabilities occur which involve perceived damages from the unlawful release (escape) of the virus. If the agent was approved for release in New Zealand, that would be a personal victory for some special interest groups but it would be an added burden for a nation that would then have to worry about a poorly understood but deadly new disease agent intentionally introduced into it's unique ecosystem, with no means of control, nor method of predicting the eventual outcome. This would be a journey into a potentially dangerous and uncontrollable aspect of the future which New Zealand can still choose to avoid.

This open letter will focus on the reported information that relates to RHD safety (host specifically for a single species) and the legitimate inferences that can come from this information. Appended will be a series of submissions previously sent to scientists, reporters, official agencies and elected officials including the Prime Ministers of Australia and New Zealand. These appendixes provide additional detail on the plasticity of caliciviruses the implications of the hemorrhagic factor virus spread, vaccine problems and societal impacts.

(a)     The RBAG executive officer states the following in the RBAG Rabbit Calicivirus 
Disease Information Kit:  "RBAG is NOT an advocate for the introduction of RCD.  
It was formed to ensure that a wide cross-section of interest groups are able to ask the hard 
questions and to ensure all necessary research was undertaken and comunicated widely."  
Having said this, the RBAG kit then proceeds with a barrage of disinformation, and wrong 
answers borrowed from the British BSE cover-up and the ANZCRDP public 
relations/misinformation mill in Australia.  The obvious intent of RBAG is to misinform the 
decision makers and public alike for the purposes of gaining approval for RHD as a 
biological agent for killing rabbits regardless of the risks involved.  RBAG is an advocate 
for the introduction of RHD.  Reviewing their "slick" pro-RHD literature confirms this.  
For RBAG to insist they are not a pro-RHD advocate group seeking to import 
and release RHD/RCD in New Zealand becomes an obvious falsehood and immediately 
opens for question any credibility that RBAG might have hoped for.

There is a virtual certainty that the RHD agent IS NOT single host species specific. The evidence for this needs to be carefully spelled out and then examined using all available relevant information. The validity of this evidence must be considered from a sound scientific point of view rather than from a politically biased viewpoint.

To begin this process, what is the evidence regarding the host range of RHD? For RHD to be declared an agent that will only infect a single species each of the following questions must be answered in the absolute negative:

1. Has RHD ever infected any species other than (Oryctolagus cuniculus)?

2. Is it now infecting any other species?

3. Could it be expected to infect any other species?

The answers to these questions cannot be the usual "there is no evidence that" which is so frequently seen in ANZRCDP and British BSE public relations releases. Such statements are inherently deceptive unless they are qualified and carefully spell out the extent to which such "evidence" was sought. If this is not done, the statements themselves become evidence of sorts. They suggest that the information in question is not being sought, a negative answer is preferred, a deception is intended without having to actually lie or perhaps the missing evidence would provide unwelcome information. Lack of evidence can never be used to prove that RHD infects only one host species.

1. ORIGINS OF RHD: In 1984 in China something not previously described took place. A shipment of Angora rabbits from Germany arrived in China and seventy-two hours later they were dying of a "new" hemorrhagic disease. This occurrence has profound implications for the species specificity of RHD. What is known of this occurrence?

a. The disease, RHD, had not been observed or reported in Germany. It's occurrence would not go unnoticed because it kills rabbits quickly in an obvious and very bloody way. The same would have been seen in China. Previous outbreaks would have been obvious.

b. The RHD disease period is 24-72 hours from exposure to death. The Chinese shipment started dying 72 hours after arrival. Therefore within 48 hours of arrival the Angora rabbit shipment converged with some second event, triggering RHD.

c. The disease spread from the Angora shipment to Chinese rabbits. Therefore, the "new" disease, is presumed to not be of Chinese rabbit origin.

d. The most reasonable explanation for the sudden appearance and high mortality is that within 24-48 hours after arriving in China the entire shipment of Angora rabbits were exposed to a new agent from some unknown animal source either in their food, or their environment.

e. This rationale argues for a non-rabbit source of the RHD agent, and against host specificity.

f. Opposing views suggest that the agent was already present in the rabbits and mutated de-novo between Germany and the first day or two in China to give a "new" disease manifestation that exhibits all the hallmarks of a disease that has never been rabbit adapted. This is a very low probability despite the findings of antibodies to similar agents in Europe pre-existing this occurrence by decades. In fact this protracted pre-existence of agents similar to but differing from RHD argues against one such agent suddenly becoming a renegade pathogen with entirely different virulence profiles. RHD destroys the liver and causes disseminated intravascular coagulation. Both can be rapidly fatal. The simultaneous appearance of two deadly factors by mutation is not likely.

g. Another compelling argument is that if a mutation event occurred to produce a deadly virus, that event by virtue of a historicity devoid of previous occurrences of RHD in rabbits, would be an extremely rare event unlikely to occur simultaneously in more than one rabbit. Therefore as a mutation event in a single rabbit, the disease should occur in a temporal way with the point source rabbit dying first, then 1-3 days later a few more and then more, as occurs with any disease introduction into a population by a single case. Involvement of the entire shipment at about 72 hours would suggest simultaneous exposure. Numerous rabbits would not simultaneously develop a very rare mutant and that mutant would be unlikely to simultaneously develop two deadly virulence factors.

h. In summary, the known information involving the origins of RHD is sufficiently compelling that one must assume the agent originated from a non-rabbit source and for that reason one must also assume that RHD is not species specific. This cannot be overcome without proof that RHD originated entirely from rabbits. Host specificity for the European rabbit has to be proven before RHD can be approved as a biological agent. RHD cannot gain such approval unless the matter of origin is resolved in a manner that would eliminate all non O. cuniculus species as the source, or specifically prove (not imply, but prove) that the European rabbit was the sole source of the agent.

2. RHD/RCD VIRUS ACQUISITION: Subsequent to making the decision to develop RHD/RCD as a biologic agent for rabbit control in Australia, attempts were made to acquire the agent from European sources. It appears the first attempt was in Germany but reportedly failed because of reluctant German scientists. Australian officials were said to have passed this off as the Germans being uncooperative and insisting on unreasonable constraints. Next the Australians went to Czechoslovakia and successfully acquired four lyophilized ampules of RHD infected rabbit liver. Since that time, a major RHD researcher in Spain told Sydney 60 Minutes' in a televised interview that he would not have given the RHD agent to Australia for use as a biological control agent against wild rabbits. In summary, without pursuing this issue further, two major European research centers for RHD do not agree with Australia's proposed use of the virus. The stated reasons by one of the experienced senior European researchers was that the virus was too unstable, and new types had already emerged which were not controlled by current vaccines. If there are doubts that the Europeans have greater credibility and experience in calicivirus research than the Australians, simply check the peer reviewed literature where the quantities of research conducted and published differ by orders of magnitude. The lesson here for Australian decision makers is that the European RHD virologists do not agree that introducing the virus is a proper decision and this is based in part on a knowledge of the fickle and changing characteristics of calicivirus. Australians who are inexperienced in calicivirology imply that they have coordinated their RHD/RCD initiative with calicivirus laboratories worldwide. In the United States the calicivirologists with decades of experience were not contacted, however, the U.S. Plum Island Laboratory (a counterpart to AAHL at Geelong) was visited. Even there, both the laboratory head and his chief, a presidentially appointed Administrator having responsibility over all U.S. animal health control programs, stated emphatically that they would
"in no way recommend that RHD be released in Australia."

On the basis of world wide opinion, the Australian and New Zealand groups promoting the release of RHD/RCD for controlling feral rabbits are very much isolated. These pro-RHD groups may rightfully claim that U.S. and European scientists fail to fully understand their rabbit problems. While this may be true, it becomes increasingly apparent that the groups in Australia and New Zealand promoting RHD release fail to comprehend the nature of this agent which causes deadly disease, nor do they consider the potential for disaster that could come from the RHD/RCD release (escape) and it's subsequent spread. This was confirmed (at the time of writing) by the Australian government deciding not to call for an environmental impact study on RHD despite serous concerns voiced in the draft EIS statement prepared by B.J. Comans concerning environmental issues including prey deprivation of native birds and loss of food source for Aboriginal groups.

3. PURE CULTURES OF THE RHD/RCD AGENT: One of the more difficult procedures in virology is to determine beyond all doubt that one is propagating pure stocks of a virus free of any contaminating viruses. There have been monumental mistakes in this regard, for example, a live virus polio vaccine contaminated with a tumor causing monkey virus, and animal vaccines contaminated with unrecognized pathogens. In private quarters there have been suggestions that the canine parvovirus pandemic which erupted world wide almost simultaneously was due to a widely distributed dog vaccine contaminated with a parvovirus growing in the cells used to produce the live virus vaccines. If a contaminating virus is not known to exist in a cell line and methods have not been developed to demonstrate it's presence then that virus will be carried and disseminated wherever the cells are propagated. If this can happen in cells of a single type grown in sealed plastic tubes which can be manipulated with pH, growth temperatures, inhibitors and numerous other mechanisms to separate viruses on the basis of their growth characteristics, think how much more difficult (or impossible) it is to "purify" viruses that must be grown in whole animals where none of these factors can be manipulated.

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