Excel Sheet semifinal CO2 rev1 - European Commissionec.europa.eu/environment/chemicals/lab_animals/pdf/x_co2.pdf · stress and subsequent stop of oxygen supply causes final euthanasia

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Question X.Aa.

Question text:

Introduction X.a) Current provisions of Directive 86/609/EEC At the end of any experiment, it shall be decided whether the animal shall be kept alive or killed by a humane method.

X.b) Current situation in Member States

The Commission has produced guidelines on humane methods of euthanasia. Some Member States use these guidelines as such. Use of CO2 is recommended for killing certain species of animals used in experiments.

X.c) Trends and implications CO2 is one of the most commonly used method of euthanasia. It is the most convenient method for euthanasia of large number of rodents (quicker, less resources and time-consuming than alternatives). If used in optimal conditions, CO2 may also be less stressful than manipulations required for injections of physical methods. It is also highly reproducible when using appropriate equipments and sage for operators.

Establishments do not use a single method of euthanasia but rather a combination of several methods according to the type of study or the circumstances. For example, if there is a necropsy at the end of a toxicology study, the animal may be anesthetized with a suitable method, and then exsanguinated. If there is no necropsy, CO2 is used (on conscious animals), or pentobarbital over-dosage, if less than 2-3 animals are to be killed at the same time.

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Statistics:

75 respondents stated that they support the preliminary analysis.

22 respondents stated that they have no opinion in relation to this question.

122 respondents stated that they do not support the preliminary analysis or components of it. To justifydisagreement, they were asked to provide arguments, facts and figures and if possible indicate the source of information. However, a number of respondents opted for the "No" reply only to supply additional arguments in support of the preliminary analysis.

Comments received:

X.d) Problem dimension The Directive itself does not specify nor give any guidance as to the most appropriate methods of killing per type of species. According to the latest research, CO2 is aversive to all vertebrates, some species find even low (10-20% by volume in air) concentrations aversive. The Panel on Animal Health and Animal Welfare of the European Food Safety Agency has in its opinion recommended to use CO2 only when animal is first rendered unconscious via another method. Banning the use of CO2 as the sole agent would, however, have significant economical impacts due to its wide use.

X.e) Potential solutions For animal welfare reason, the revision could incorporate the list of humane methods of euthanasia to be used for experimental animals. Within the list of methods, the use of CO2 could be prohibited without rendering the animal unconscious prior to its use.

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Name of the expert or organisation

Type of organisation

Activity level of the organisation

Representative- ness

X.Aa.

Gesellschaft zur Förderung der biomedizischen Forschung

User of animals -public sector

National The organisation is representative for the field at the respective activity level.

No Ban of CO2 is scientifically not justified and has no animal welfare impact !

Hannover Medical School


Local; Regional; National

Hannover Medical School currently ranks highest with respect to research activities and aquired research grants

Ban of CO2 is scientifically not justified and has no animal welfare impact!See also FELASA comment as well as RCVS letter

Hannu Komulainen


National; European

Is relevant CO2 should not be prohibited as a stand alone method.

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Hans J. Hedrich User of animals -public sector

Local; Regional; National; European

I am currently President of the European College of Laboratory Animal Medicine (ECLAM) and Vicepresident Gesellschaft zur Förderung der Biomedizinischen Forschung

No Ban of CO2 is scientifically not justified and has no animal welfare impact!

Jose A. Costoya -Universidde de Santiago de Compostela, Spain


Local; Regional; National; European; Worldwide

Yes Certain IACUCs in USA use American Veterinary Medical Association recommendations and CO2 is considered as acceptable for several species. I believe there is some scientific controversy on this, therefore further investigations must be done before CO2 is prohibited.

Prof. Dr. R. Nobiling, University of Heidelberg


Regional; National; European; Worldwide

Biomed research at Univ. Heidelberg is Top - ranked

A undifferenciated prohibition of CO2 is scientifically not justified and has virtually no positive impact on animal welfare.

Prof.Dr. Günter Klima


Not applicable yes medical university

From my experiments I know that CO2 is very good

Timo Nevalainen User of animals -public sector

Local Yes It is not common EU-policy to exclude a single agent at a point where disagreement on effect still exists.

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(Name confidential - Respondent 001)


National; European; Worldwide

yes According to our experience, there is no scientific reason for a ban of CO2 euthanasia if done properly.




XXX The argument is fundamentally flawed and lacks evidence - see justification.




Yes A recent workshop in the UK has concluded that other gaseous anaesthetics produce similar behavioural effects to CO2. In addition to the unacceptable effects of the alternative gases they additionally carry significant cost to introduce and use.



Worldwide there are others Ban of CO2 is scientifically not justified and has no animal welfare impact



National representative I do not think that prohibition to use CO2 without rendering the animal first unconscious by a use of another method, or by use of anaesthetic gases in combination with CO2 will greatly increase animal welfare, because there are data contradicting the assessment euthanasia with CO2 being inhuman (Lab Anim 34:91, 2000).



National jes -

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National Yes, we are representing all physiological researchers in the Netherlands; we have about 300 members

The opinions that CO2 is aversive and poorly effective are wrong. It is well known that initially starting the procedure with a mixture of high CO2 and oxygen sedates the animals without major stress and subsequent stop of oxygen supply causes final euthanasia. Halothane should absolutely be discouraged bue to the risks involved to the personnel (halothane has almost been abandoned in human anesthesia for this reason).



Worldwide One of several global pharmaceutical companies

It is difficult to understand why such a technical detail should become part of a directive. There will be always a certain, usually low level of stress exposure. Most, if not all anaesthetics can cause excitation to varying degrees under certain circumstances. I wonder whether the EFSA report is scientifically tenable. There are studies showing no increase in stress hormones when the certain precautions are taken.




Other organisations in Austria: Karl-Franzens-University; Medical University Vienna; Medical University Innsbruck

See 1.4



Local Yes CO2 euthanisation is fast and very effective for all laboratory animals, and does not causes significant stress. There is no scientific proof that there is a need to render animals unconscious before euthanisation by CO2. In exceptional cases it may be useful to render larger animals than rodents unconscious before use of CO2. This could become a recommendation for exceptional cases.

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Yes CO2 is not inhumane and in certain experiments cannot be replaced by other means.




Yes Findings by the University of Newcastle do not support the above statement that CO2 should only be used once an animal is unconscious



National yes Recent research by the Uk National Centre for the three r's has concluded that the main alternative methods to CO2 are no more humane, therefore I see no need to ban it's use on conscious animals.



Not applicable equivalent to other large medical universities

In rodents, rising levels of CO2 provide unconsciousness prior to asphixiation (see Flecknell, 2006) CO2 euthanasia requires little or no handling of rodents and is there less stressful than injection/physical methods of euthanasia. Rodents resent being restrained.



Regional yes see 1.4



Not applicable yes The stress of transportation to an euthanasia setup usually outweighs the short (few seconds) effects of CO2

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Local; National There are several central laboratory animal facilities at Austrian universities

If done properly there is no scientific reason for a ban of CO2 euthanasia.



National; Worldwide

There are other universities at the respective activity level

No, it has not yet unequivocally been proven that there are more humane methods that can be used for euthanasia compared with CO2. A report from a recent workshop on CO2 euthanasia, soon to be published by the UK National Centre for the 3Rs, concludes that it is premature to ban it as there is no practical alternative that could be said to be more humane



National yes. There are other academic medical centres

There is conflicting data in literature. The statement that CO2 is aversive to all vertebrates should, therefore, not be made. Additional research is needed in order to come recommendations that have a scientific basis. Furthermore, it is not consistent with the nature of the Directive to single out one technique. The emphasis should be on the intention that techniques and procedures in general should be state-of-the-art.



Local yes No, if CO2 is used in a proper way, it is still a good method (for details see Dissertation Silke Corbach: “Untersuchung der CO2-Euthanasie bei Labormäusen auf Tierschutzgerechtigkeit” - “Investigation of Carbon Dioxide-Euthanasia of Laboratory Mice Regarding Animal Welfare Aspects”; http://elib.tiho-hannover.de/dissertations/corbachs_ss06.pdf).



Local yes No, if CO2 is used in a proper way, it is still a good method (for details see Dissertation Silke Corbach: “Untersuchung der CO2-Euthanasie bei Labormäusen auf Tierschutzgerechtigkeit” - “Investigation of Carbon Dioxide-Euthanasia of Laboratory Mice Regarding Animal Welfare Aspects”; http://elib.tiho-hannover.de/dissertations/corbachs_ss06.pdf).

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Yes Not sufficient evidence to reach this conclusion - not based on fact




Our organisation is representative within the field of cancer research but we have many collaborations with other like organisations.

Sufficient research needs to be done to provide factual information on the use of carbon dioxide for animal euthanasia. Many of the present arguments against the use of this agent and not factually-based and can be misinterpreted.



National yes The use of CO2 for euthanasia under certain circumstances can induce distress in rodents (ACLAM task force on rodent euthanasia 2005). It is still unclear if it the use of CO2 or the concentration of CO2 (70% vs 100%) or the rate of filling gas chamber (prefilled chamber versus low rate or high rate). More scientific information is still needed to define guidelines for the use of CO2. The cost evaluation do not consider extra cost for halothane and isoflurane use and for the maintenance of scavenging.




yes, we are a major EU HEI for biomedical research

The present directive is sufficient. It is a technical matter whether euthanasia by increased CO2 is sufficiently humane, in comparison with other available methods. the proposed use of halothane has significant health and safety implications for human personnel.



National Some smaller institutes exist in the Netherlands

No. The negative effects for animals of properly executed CO2 euthanasia are still subject of debate. There is by no means consensus about its being less humane than other accepted forms of euthanasia, when properly executed.

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National yes Ban of CO2 is scientifically not justified and has no animal welfare impact



Local organisation is representative

this question was not in the questionnaire when I printed it out




no If CO2 is derived from gas tanks, and the cage of animals is slowly filled, these animals do not show any symptoms of stress or pain. The use of CO2 has to be reglemented, but not prohibited. Solutions for this problem were provided by the NIH in conjunction with the american veterinary board and could be directly applied in Europe.

Bayer HealthCare

User of animals -private sector

Worldwide organisation is representative for the field

http://by-ej-sid.bayer-ag.com/han/EJ-1648/www.nature.com/nature/journal/v441/n7093/full/441570a.html

GlaxoSmithKline User of animals -private sector

Worldwide Thre are other international pharamceutical companies. We produce 25% of the worlds vaccines

These assumptions are fundamentally flawed. The researchers who work in this area do not support this call for a ban. http://www.nc3rs.org.uk/news.asp?id=292 Neither does the senior scientific community http://www.royalsoc.ac.uk/news.asp?id=5000

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Protherics Medicines Development Ltd.


Worldwide No - ABPI and EFPIA repesent the pharmaceutical industry in UK/EU

EFSA report is inaccurate so it must not be used to set policy. Alternative methods for rodents carry similar or great risks: gaseous anaesthetics produce similar behavioural effects to CO2 at induction concentrations, inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Scientific literature on current CO2 issues is on rats, not mice and other rodents. The important impact of age is not addressed, so laws would have to be species-specific, they would also have to be age specific. The inclusion of technical minutiae about methodologies is not consistent with the purpose of a Directive and will limit the scope for the future refinement of methodologies as new scientific data emerge. Financial costs would be very much higher than stated with uncertain science and no clear alternatives to CO2.

sanofi pasteur R&D


Worldwide Vaccine field No scientific consensus on the detrimental effects of CO2, when well used. But Yes, for the costs associated to the change: increase man-day time, increase equipments…

SYNGENTA User of animals -private sector

Worldwide There are other relevant organisations

Research presented at British Laboratory Animal Veterinary Association Spring Meeting in March 2006. Unpublished research showing the use of slow fill CO2 and the loss of conciousness by rats before any responses to pain were detected and nociceptors activated. Therefore there is the possibility of using CO2 in a humane manner. Gaseous anaesthetics can also be aversive.




Sanofi aventis is the largest pharmaceutical company in Europe and ranks third worldwide

The EFSA report is scientifically inaccurate and has ignored currently available data. It is misleading and biased See: ACLAM Task Force on Rodent Euthanasia, see also work being done by Flecknell and Roughan.

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Worldwide Yes There are broader issues to be addressed before considering the specifics of CO2. The Commission has produced guidance on humane methods of euthanasia - should it incorporate these into the Directive or should they remain as guidance? Given that the science moves on quite quickly, we would support guidance that is reviewed regularly, rather than inclusion in the body of the DIrective. Specifically with respect to CO2, we cannot conclude that the EFSA report is balanced and comprehensive in its review of this topic.



Worldwide Research and development

no comment



Worldwide Yes The issue should be discussed elsewhere. More research in this field is needed.



Worldwide yes it's representative

Recommendations already exists for euthnasia (AVMA panel, AAALAC, …) and they are references. CO2 is not prohibited in these texts and anesthesia before euthanasia with CO2 is already a recommendaiton in these texts.



National; Worldwide

yes The EEFSA should not be used as a unique or dominant reference. It can be challenged scientifically and by practical experience. . For rats and mice CO2 euthanasia is a humane and very effective technique IF used in the right way (like any sensitive technique) Other methods usable for rodents do not present any advantage and also have to be used in a specific way (gaseous anaesthetics, argon, nitrogen…) and may even be less effective and less "ethical". CO2 euthanasia is an excellent technique when used professionally i.e. with a progressive increase of the CO2 concentration, a flow and distribution control in a suitable chamber, time control of injection in the chamber and exposure, etc. One exception is that it shouldn't be used for immature rodent pups or with a very long CO2 exposure time. It is not the purpose of a directive to address such detailed technical issues

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European Representitive In my experience it is very important to get the flow rate at which the CO2 is delivered correct, for a rapid induction and death. Too fast I agree is aversive, and too slow causes suffering. at about 20% of the volume to be filled per minute induces unconciousness rapidly, and brings about a humane death.



Worldwide other relevant organisations

- The list of human method of killing should not be included in the Directive , publications and reports of experts are available and new methods can occur before revision of the directive - Technical and scientific data on CO2 in rodents must be take into account and practical use (Good practice for use of CO2) must be defined (different advices in the literature) - Halothane is hazardous



Worldwide yes No increase in animal welfare by adding an additional anaesthesia when CO2 application is carried out by trained persons



Worldwide There are many other such organisations in Europe

The above analysis does not take account of how little technical information has been published by a peer review process in this field. For example, there is no information offered on mice. The gaseous anaesthetic agents being proposed as alternatives are also aversive. The proposal is of concern in that, if enacted in legislation, it may unintentionally encourage reversion to physical techniques of killing or the use of injectable agents being performed on large groups of animals. If the legislation was enacted, the proposals would have to be highly detailed in terms of species and age or development stage, and this appears undesirable technical detail for proscription. As long as there is no scientifically validated consensus on an alternative to the use of CO2, it appears premature to single out this technique for banning.



Worldwide No I believe that the EFSA report is flawed in relation to use of CO2. If used correctly it is a safe and acceptable method for euthanasia of rodents.



Worldwide pharmaceutical since a list of humane euthanasia methods is subjected to further research knowledge, changes must remain flexible and should be incorporated in recommendations or guidelines from e.g. FELASA, but not made rigid in the Directive.

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Worldwide Yes We do not agree that avoidance of the use of carbon dioxide for euthanasia will increase animal welfare. A test by 2 of our personnel and observed by 3 official representatives (NVS, NACWO and dNACWO) used CO2 introduced at 2 L/min = 20% chamber vol/min, 4 L/min = 40% of chamber vol/min and 6 L/min = 60% of chamber vol/min. The 2 lower flow rates resulted in protracted behavioural adverse effects in the rats that could act adversely on animal welfare. This response was reduced by using a flow rate of 6 L/min (60% of chamber vol/min). Importantly at this increased rate, no inappropriate effects were observed.



Worldwide Global pharmaceutical research and development

Ban of CO2 is scientifically not justified - Alternative methods for rodents carry similar or great risks: gaseous anaesthetics produce similar behavioural effects to CO2 at induction concentrations, inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Scientific literature on current CO2 issues is on rats, not mice and other rodents. The important impact of age is not addressed, so laws would have to be species-specific, they would also have to be age specific. The inclusion of technical minutiae about methodologies is not consistent with the purpose of a Directive and will limit the scope for the future refinement of methodologies as new scientific data emerge. Financial costs would be very much higher than stated with uncertain science and no clear alternatives to CO2.



Worldwide yes EFSA report=scientifically inaccurate: must not be used to set policy (see P.Flecknell, ESLAV meeting Oct 05, ESP). Alt. methods for rodents carry similar or great risks: gaseous anaesth. produce similar behavioural effects to CO2 at induction concentr.,inert gases like argon lead to signs that require investigation and nitrogen is ineffective.No clear opinion in scientific community if use of CO2 euthanasia (E) has major neg. impact in animal welfare (AW).As long there is no comm..accepted opinion that CO2 (E) affects AW negatively,this shouldn’t be included in a Directive.Scientific literature on current CO2 issues is on rats,not other rodents.Important impact of age=not addressed,so laws would have to be species-&age specific. inclusion of tech. minutiae about methodologies=not steady with purpose of a Directive&limits the scope for future refinement of methodologies as new scientific data emerge.Costs much higher than stated with uncertain science&no alternatives to CO2.

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there are multiple independent Contract Research Organisations in UK and EU

The administration of halothane or isoflurane before killing animals by inhalation of CO2 will negate most of the advantages of using CO2. e.g. when testing for embryo-foetal toxicity pregnant rats are killed by CO2 to avoid anaesthetic agents affecting the viability of the unborn pups



National Yes The report provided by EFSA does not meet the standards and requirements for a scientific report, in particular if this report is founding policy making. Alternative methods for euthanasia of rodents have similar risks as gaseous anaesthetics produce the same behavioural effects to CO2 at induction concentrations and inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Costs will be higher than stated.



Worldwide Representative Banning of CO2 for euthanasia is not based on real scientific evidence, and the EFSA Report itself does not consider the most recent scientific data.



Worldwide yes Some of the conclusions of the EFSA report are the subject of scientific disagreement especially on CO2, so it must not be used to set policy. It does not make sense for the Directive to detail acceptable methods. Science, technology and scientific opinion move on and this would be counterproductive to animal welfare as it would hinder the adoption of improved methods in the future. A framework regulation requiring methods consistent with current best practice would be a much better solution then listing methods that are currently considered to be acceptable.

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Worldwide yes Alternative methods for rodents carry similar or great risks: gaseous anaesthetics produce similar behavioural effects to CO2 at induction concentrations, inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Scientific literature on current CO2 issues is on rats, not mice and other rodents. The important impact of age is not addressed, so laws would have to be species-specific, they would also have to be age specific. The inclusion of technical minutiae about methodologies is not consistent with the purpose of a Directive and will limit the scope for the future refinement of methodologies as new scientific data emerge. Financial costs would be very much higher than stated with uncertain science and no clear alternatives to CO2.



Worldwide no · no 100% clear data about aversive effects or not of CO2 or other anaesthetic gases. · CO2 may cause aversive effects (human data) in some rodent species · Anaesthesia of animals before use of CO2 is desirable



Worldwide The bquestionaire will be completed for the German sites. Other site head in other countries will reply individually

There is no scientific agreement on the use of CO2 so far.

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Worldwide Many organisations

The EFSA report is scientifically inaccurate so it must not be used to set policy. The current consensus from all active researchers is at: (LINK). Alternative methods for rodents carry similar or great risks: gaseous anaesthetics produce similar behavioural effects to CO2 at induction concentrations, inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Scientific literature on current CO2 issues is on rats, not mice and other rodents. The important impact of age is not addressed, so laws would have to be species-specific, they would also have to be age specific. The inclusion of technical minutiae about methodologies is not consistent with the purpose of a Directive and will limit the scope for the future refinement of methodologies as new scientific data emerge. Financial costs would be very much higher than stated with uncertain science and no clear alternatives to CO2.



Worldwide Yes There a no humane alternatives to appropriate (high concentration, rapid application) use of CO2 for euthanasia of large numbers of rodents.



Worldwide Yes Since your data already supports the fact that there is no significant improvement to be gained.



National representative http:// www.nature.com/nature/journal/v441/n7093/full/441570a.html



Worldwide There are any other relevant organisations but we are making research with rodents

The use of C02 couldnot be prohibited If there is suitable not a new method

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Worldwide Don't understand this question

There are broader issues to be addressed before considering the specifics of CO2. The Commission has produced guidance on humane methods of euthanasia - should it incorporate these into the Directive or should they remain as guidance? Given that the science moves on quite quickly, we would support guidance that is reviewed regularly, rather than inclusion in the body of the DIrective. Specifically with respect to CO2, we cannot conclude that the EFSA report is balanced and comprehensive in its review of this topic. This appears also to be the opinion of the Royal Society: http://www.royalsoc.ac.uk/displaypagedoc.asp?id=21379

Animal Procedures Committee (APC)

Public authority National The APC provide independent advice to UK Government on the use of animals in scientific procedures. The APC consider science, industry and animal welfare. Members are from a variety of backgrounds.

The APC does not support regulating euthanasia methods at this level of detail.

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Coordination group for laboratory animal activities

Public authority National Yes It is not common EU-policy to exclude a single agent at a point where disagreement on effect still exists.

DR Achim Klug, Ludwig Maximilians University Munich

Public authority Worldwide; Not applicable

Deutsche Forschungsgemeinschaft - DFG

Many alternatives to Co2 are available and are currently used. The method of killing is already included in current permits and reviewed by competent veterinarians before permits are issued, and thus no further regulation is required.

Prof. Dr. Bernd Hoffmann, Justus-Liebig-Universität Giessen

Public authority National; European

similar to other universities

The question is valid, the proposal not. Further research is reqiured, accounting for mor species than the rat. Also dogs, cats, pigs etc are used for experimental puposes.

UK Medical Research Council

Public authority Local; Regional; National; European; Worldwide

The MRC is the UK's leading publicly funded biomedical research organisation

Please refer to section 1.4 below.


Public authority National Yes EFSA advice has not been peer reviewed, but is selective and incomplete. Alternatives proposed likely to have similar issues.

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Public authority Local yes Ban of CO2 is scientifically not justified and has no animal welfare impact




Public authority National Yes See 1.4


Public authority National Regulatory authority in UK

Unresolved scientific issues mean that this assessment is made from incomplete knowledge in the area and some conclusions could be invalid or may alter later (see also 1.4)


Public authority National; European; Worldwide

Although we are a smaller University, about half of the research groups involving animal experiments belong to leading institutions in their field.

see comment 1.1

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Public authority National representative Ban of CO2 is scientifically not justified and has no animal welfare impact !

1.Animal Aid Other National Other organisations do exist

While we support the compilation of a list of humane methods of euthanasia to be used for animals used in experiments, the use of CO2 alone is unacceptable - whether administered under ‘optimal conditions’ or not.

Biosciences Federation

Other National; European

There are other relevant organisations

There is not yet a consensus about the welfare benefits of banning CO2 euthanasia. See details of a recent workshop on the subject (via: www.nc3rs.org.uk/news.asp?id=292).

COST Action B24 “Laboratory Animal Science Welfare” - http://biomedicum.ut.ee/costb24

Other European No No, if CO2 is used in a proper way, it is still a good method (for details see Dissertation Silke Corbach: “Untersuchung der CO2-Euthanasie bei Labormäusen auf Tierschutzgerechtigkeit” - “Investigation of Carbon Dioxide-Euthanasia of Laboratory Mice Regarding Animal Welfare Aspects”; http://elib.tiho-hannover.de/dissertations/corbachs_ss06.pdf). Singling out one technique is neither consistent with the nature of the Directive.

Medical University of Vienna

Other Local; Regional; National; European; Worldwide

Research in Medicine

see 1.4

National Centre for the Replacement, Refinement and Reduction of Animals in Research

Other National Representative More research is required before deciding to ban use of CO2 euthanasia. See Newcastle consensus report at www.nc3rs.org.uk

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Other National I am the chair of the Hungarian Scientific Ethical Committeeon Animal Experimentation

To review and update the list of appropriate euthanasia methods is a scientific task and not a legal job. It has no place in the revised Directive. The appropriateness of CO2 euthanasia is a scientific matter so let the scientists discuss it and do not intervene by legal means.


Other Not applicable other According to our experience, there is no scientific reason for a ban of CO2 euthanasia if done properly.


Other National Yes N.B. It is of great concern that this section is based on the EFSA report, which is scientifically inaccurate and should therefore not be used as a basis for policy. We therefore disagree with the findings of this section of the questionnaire and has not responded to the remaining questions in this section. If this is not addressed and the policy making on the review of Directive 86/609 continues to be based on inaccurate science, we are extremely concerned about the detrimental impact that this could have on the emerging bioscience sector in Europe, which the European Commission has done so much to support.


Other Not applicable not applicable This issue should be discussed elsewhere. I do not believe that it belongs into the Directive, since some of the problems may result from insufficient methodology. However, more research is needed in this field.

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Other Worldwide The Institute of Animal Technology is the prfessional body representing animal technologists

The Institute of Animal Technology is concerned about the proposal that CO2 is an unsuitable agent for the euthansia of rodents. Our members are required to euthanase animals as part of their employment and in the UK are the largest group most likely to euthanase animals in the laboratory environment. Feedback from our membership is that actual experience indicates that CO2 administered correctly in a rising concentration, with initial induction from a normal air environment, does not compromise animal welfare. The Institute is aware of the debate surrounding the use of CO2 and its welfare implications. However, we are not aware of any definitive agreement that any practical alternative is more humane and indeed other alternative agents may pose a greater health and safety risk to our members.

Austrian Society of Toxicoloy (ASTOX)

Non-governmental organisation

National Yes see 1.4

European Biomedical Research Association


European Yes There is not yet a consensus about the welfare benefits of banning CO2 euthanasia.

Gesellschaft für Versuchstierkunde - Society for Laboratory Animal Science (GV-SOLAS)


National; European

Yes No, if CO2 is used in a proper way, it is still a good method (for details see Dissertation Silke Corbach: “Untersuchung der CO2-Euthanasie bei Labormäusen auf Tierschutzgerechtigkeit” - “Investigation of Carbon Dioxide-Euthanasia of Laboratory Mice Regarding Animal Welfare Aspects”; http://elib.tiho-hannover.de/dissertations/corbachs_ss06.pdf).

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The Royal Society


National Yes. The Royal Society is the independent scientific academy of the UK dedicated to promoting excellence in science. The Society plays an influential role in national and international science policy

See justification below

UK Laboratory Animal Science Association (LASA)


National LASA represents animal user community in academic, government & industry sectors.

The EFSA report is scientifically flawed and should not be used to set policy. A recent workshop in the UK has concluded that other gaseous anaesthetics produce similar behavioural effects to CO2. In addition to the unacceptable effects of the alternative gases they additionally carry significant cost to introduce and use. The evidence for the adverse effects has been mainly generated in rats whilst probably 80% of all rodents that are euthanased by this method are mice.

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European FEPS is an umbrella organization for national Physiological societies in Europe. See further http://www.feps.org

I (Hultborn) have personally participated on a panel to judge CO2 anesthesia several years ago (Denmark – slaughtering of pigs) – I do not really recognize/ remember the conclusions referred to here. I would also like to refer to the comments made both by EBRA (see EBRA Bulletin June 2006; European Biomedical Research Association) and the Royal Society on this point.



National yes There exists no scientific reason for a ban of CO2 euthanasia if done properly.



National NO OTHER RELEVANT ORGANISATION

Despite its widespread use, CO2 euthanasia is not standardized (see Report of the ACLAM Task Force on Rodent Euthanasia, August 2005). This lack of standardization may be responsible for bad performance. The first step should be to set the technical requirements for this method of euthanasia. There is no worldwide scientific agreement regarding the aversive effect of CO2 and new data become regularly available (see JAALAS, 45, 4, July 2006, page 78, PS20).



National YES The aversiveness of Co2 needs to be further investigated.



National Represents all Finnish pharmacologists

See below

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National The Danish Society of Pharmacology and Toxicology represents Danish pharmacologists and toxicologists.

See 1.4.



Worldwide No other organisations

We understand from the report of the Newcastle Consensus Meeting on Carbon Dioxide Euthanasia of Laboratory Animals (http://www.nc3rs.org.uk/downloaddoc.asp?id=416&page=292&skin=0) that whilst the use of CO2 is aversive to some species of animals; there is a need for further research before any type of prohibition is implemented, as there is insufficient information currently available on alternatives to CO2. Therefore, any proposed ban on the use of CO2 is premature.



Regional Yes see 1.4




One other The experimentation on animals is not the only one and is obsolete surpassed by others more trustworthy.



Worldwide - -

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National yes The statement of the Panel on Animal Health and Animal Welfare of the European Food Safety Agency about the CO2 method for rodents is scientifically inaccurate. A ban or recommendation of additional anaesthesia in combination with CO2 is unjustifiable. Other gaseous anaesthetics show similar behavioural effects at induction concentrations Inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. The inclusion of technical details in methodologies is not consistent with the purpose of a Directive and will limit the scope for future refinement of methodologies as new scientific data emerge. Additionally the costs will increase disproportionate.



Worldwide yes Ban of CO2 is scientifically not justified and has no animal welfare impact



Worldwide The field at the respective activity level

Is very important to have validate methods of eutanasia with CO2. Is difficult ( security of people and envirommental )to have the god method for have unconscious prior the use of CO2


Breeder of animals

Worldwide We are representative for the field and constitute about 50 percent of the activity

Recent studies conducted in the UK and supported by work done in Canada and the US do not support Items 1.1, 1.2 or 1.3. In addition, these Items do not take into account that certain anesthetic gases such as Halothane are being phased out of the marketplace in favor of more difficult to administer and more costly anesthetic gases used principally for humans. The Items do not take into account the hazards associated with such gases especially to women of childbearing age, and from our estimation, greatly underestimates the costs per institution especially for very large institutions where one would expect not only the costs for operation but the costs for acquiring many pieces of such equipment would be very substantial. These Items also do not recognize the fact that the gases in question, when administered in a chamber setting, are also aversive to rodents and produce at the very minimum a stressful environment.

28

Biotechnology and Biological Sciences Research Council

National Representative There is an extensive discussion of evidence concerning the welfare impact of carbon dioxide applied using different methods but this evidence is controversial and conflicting. Notably, research carried out in the laboratory of the Chair of the working group that developed the recommendations upon which this consultation is based concludes that carbon dioxide is more aversive than a range of inhalation anaesthetics, but other researchers have not come to the same conclusions.


Regional; National

Not applicable: response is personal opinion based on personal expertise

Please see arguments below with respect to prohibiting the use of CO2 without pre-stunning. A list of approved humane methods of euthanasia to ensure animal welfare is a good idea - however, it is essential that it can be updated easily in the light of new research such that better methods can be added simply and equally, methods for which evidence becomes available indicating them to be sub-optimal can be removed.


National; European

There are other organisations

See report at www.nc3rs.org.uk/news.asp?id=292


National There are other relevant organisations

more research is required before deciding whether to ban the use of CO2


Local representative The statement of the Panel on Animal Health and Animal Welfare of the European Food Safety Agency about the CO2 method for rodents is scientifically inaccurate. A ban or recommendation of additional anaesthesia in combination with CO2 is unjustifiable. Other gaseous anaesthetics show similar behavioural effects at induction concentrations Inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. The inclusion of technical details in methodologies is not consistent with the purpose of a Directive and will limit the scope for future refinement of methodologies as new scientific data emerge. Additionally the costs will increase disproportionate.

29



Yes, we are representarive for the field

The statement of the Panel on Animal Health and Animal Welfare of the European Food Safety Agency about the CO2 method for rodents is scientifically inaccurate. A ban or recommendation of additional anaesthesia in combination with CO2 is unjustifiable. Other gaseous anaesthetics show similar behavioural effects at induction concentrations Inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. The inclusion of technical details in methodologies is not consistent with the purpose of a Directive and will limit the scope for future refinement of methodologies as new scientific data emerge. Additionally the costs will increase disproportionate.

30

Question X.1.1.

Question text:

Statistics:




Comments received:





X.1.1a.

The overall preliminary assessment shows that prohibition to use CO2 without rendering the animal first unconscious by a use of another method, or by use of anaesthetic gases in combination with CO2, would greatly increase animal welfare but would also be associated with higher costs.

Overall assessment:

Do you support this overall analysis? Yes/No/No opinion

31

Dept. Laboratory Animal Science, Utrecht University, The Netherlands


Worldwide chair of lab. animal sci. recognized by national authorities/responsable for national coordination education and training in lab. animal sci./ leading role in international laboratory animal science

Very few research results demonstrate the aversiveness of CO2 and, in particular, the statement that CO2 is aversive to “all vertebrates” is an exaggeration. On the contrary, many other publications demonstrate its usefulness for euthanasia. Furthermore, recent studies by Flecknell et al have not been taken sufficiently into account in formulating this assessment..





32





Ban of CO2 is scientifically not justified and has no animal welfare impact !




No, if CO2 is properly used, it is still a good method (for details see Dissertation Silke Corbach: “Untersuchung der CO2-Euthanasie bei Labormäusen auf Tierschutzgerechtigkeit” - “Investigation of Carbon Dioxide-Euthanasia of Laboratory Mice Regarding Animal Welfare Aspects”; http://elib.tiho-hannover.de/dissertations/corbachs_ss06.pdf).

Prof Alan Palmer


Worldwide Yes The state of present knowledge is not sufficient to justify an outright band on the use of CO2 at present.

33







Local Yes It is not common EU-policy to exclude a single agent at a point where disagreement on effect still exists.

Torgny Jeneskog, Umeå University, Sweden



Yes, guess so ... There are some - but not overwhelming, and also contradictory - data to support an adversiveness for CO2-inhalation in some species. BUT the adversiveness of a certain agent/method has to be put in relation to the adversiveness of other possible agents/methods. For many (most?) of those, including "anaesthetic gases", not much data are available to support their supremecy over CO2. Physical methods and injection techniques (overdose of anaesthetic) are indeed "stressful", per se, to the extent that the adversiveness of CO2-inhalation might in fact be negligable. [This might be arguments in general for more scientifically based studies on euthanasia methods!]








XXX See 1.4




Yes It is not clear that there are alternative options that are more humane when dealing with groups of animals. A ban at this stage would be premature.

34



National XXX are representative of our scientifi activity (basic research)











as mentioned above, well used CO2 euthanasia is not aversive




It is difficult to understand why such a technical detail should become part of a directive. There will be always a certain, usually low level of stress exposure. Most, if not all anaesthetics can cause excitation to varying degrees under certain circumstances. I wonder whether the EFSA report is scientifically tenable. There are studies showing no increase in stress hormones when the certain precautions are taken.

35





See 1.4



Local Yes There is no scientific proof in recognized literature that animal welfare would greatly increase by rendering animals unconscious before CO2 euthanisation. It is highly doubtful that this would be the case because CO2 euthanisation is very fast and effective.




Yes see 1.4



European Yes A recent study soon to be published in the UK National Centre for the Three Rs will conclude that alternative methods of euthanasia are not significantly more humane than CO2. The present assessment recognised the initial one-off extra costs of alternatives, but not the additional ongoing costs in terms of time and manpower per animal euthanased. If alternatives do not provide clear additional benefits it will be difficult to justify the additional costs.




CO2 euthanasia when conducted properly with rising levels of CO2 does not cause distress or suffering to rodents (see Flecknell, 2006).

36



Regional yes see 1.4







see X.e. There is no need for a combination with other anesthetic gases.



National; Worldwide






See comments made in Aa introduction



Local yes see above X.e

37



Local yes see Aa.




Yes As above. Not enough evidence to support this - more research is needed. Any current evidence is anecdotal.



National yes The use of CO2 for euthanasia under certain circumstances can induce distress in rodents (ACLAM task force on rodent euthanasia 2005). It is still unclear if it the use of CO2 or the concentration of CO2 (70% vs 100%) or the rate of filling gas chamber (prefilled chamber versus low rate or high rate). More scientific information is still needed to define guidelines for the use of CO2.




No. The negative effects for animals of properly executed CO2 euthanasia as compared to other forms of euthanasia are still subject of debate.







no If CO2 is derived from gas tanks, and the cage of animals is slowly filled, these animals do not show any symptoms of stress or pain. The use of CO2 has to be reglemented, but not prohibited. Solutions for this problem were provided by the NIH in conjunction with the american veterinary board and could be directly applied in Europe.

Bayer HealthCare



1.1 As above, we reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk. http://by-ej-sid.bayer-ag.com/han/EJ-1648/www.nature.com/nature/journal/v441/n7093/full/441570a.html

38



as above




We reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk. The second paragraph is not relevant. Statement that CO2 is only used in toxicology studies where there is no necropsy (Section X c) is incorrect – many laboratories routinely use CO2 to euthanise rodents for toxicology studies, as it is quick and effective, and the method does not produce any significant changes in the tissues that could interfere with the pathology evaluation. Any change in methodology for toxicology studies needs careful evaluation, to avoid introducing artefacts into the histopathological assessments, as a result of effects of agents used for euthanasia - halothane and phenobarbital both produce tissue changes that could interfere with the interpretation of toxicity studies.

sanofi pasteur R&D


Worldwide Vaccine field No scientific consensus on the detrimental effects of CO2, when well used. But Yes, for the costs associated to the change: increase man-day time, increase equipments…



National yes There are no alternatives and the costs and the risk to bring the procedure into the opposite direction (more stress, less welfare) could occur.

39





I reject the proposal on scientific grounds. It uses the EFSA report as its base and this misrepresents the current data as well as ignores data that was made available at the time of writing the report. Current data (Flecknell et al.) show that CO2 euthanasia is an appropriate method if used correctly. See also : http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstract&list_uids=16542038&query_hl=3&itool=pubmed_docsum



Worldwide Yes The evidence on the effects of CO2 on welfare are uncertain and hence we cannot support the analysis that there would be a great increase in animal welfare. However, we do support the view that there would be higher costs of alternative methods.




no comment



Worldwide Yes The issue should be discussed elsewhere. More research in this field is needed.




CO2 is one of the best method for euthanasia. The gaz that could be used with CO2 is mainly O2 or normal air and so, increasing CO2 concetration in a box with animals breathing normal air will lead to anesthesia before euthanasia. No need to change



National; Worldwide

yes See previous comment above. This statement cannot be accepted both for scientific, ethical and practical reasons.




See above

40



Worldwide yes No great increase in animal welfare by adding an additional anaesthesia when CO2 application is carried out by trained persons




The analysis cannot be supported as there is an assumption that other gaseous agents lack the aversion of CO2 and this is not supported by the available scientific information. Therefore there may not be a greatly increased impact on welfare. The banning of the technique might encourage more animals to be killed by direct means of handling to administer injection or a physical method.



Worldwide No I don’t agree that there is a large welfare gain to be had from removal of CO2 as a euthanasia agent. Different species find CO2 aversive at different concentrations, and the latest work by Flecknell and others indicates that when rats are exposed to a rising concentration of CO2, they are rendered unconscious before concentrations of CO2 are reached which would be considered painful to them, as judged by physiological measurements. Removal of CO2 would undoubtedly lead to higher costs due to the need to find a sutiable and safe alternative.



Worldwide pharmaceutical CO2 euthanasia is, as all gaseous methods are, of course aversive, but also acceptable as other methods under certain usage conditions. It differs according to the species (mice are becoming unconscious rapidly and with no apparent stress), the age of the animals, whether the induction box is prefilled with CO2 or gradually filled (ref.: Paul Flecknell showed at the ESLAV-SECAL meeting in Elche Oct 5-7, 2005, that the rat become unconscious BEFORE showing signs for stress when the induction box is filled at a flow of 20% of the box volume/minute) and probably other factors. The EFSA report, being only one of the many literature reports that show different results because experiments were done in different conditions, should not be used as sole reference. Rather additional research is needed like also encouraged by the ICLAS (Science May 5, 2006). Future refinement of methodologies is needed as new scientific data emerge



Worldwide Yes Use of a gaseous anaesthetic prior to exposure carbon dioxide will be very costly and we do not agree that this will increase animal welfare. We find that carbon dioxide, when administered at a carefully controlled flow rate, is a humane method of killing rodents.

41



Worldwide YES (Safety Assessment)

As above, we reject the policy proposal on scientific grounds: welfare effects are very questionnable and the costs would be high.



Worldwide yes As above, we reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstract&list_uids=16542038&query_hl=3&itool=pubmed_docsum The second paragraph is not relevant.








National Yes The report provided by EFSA does not meet the standards and requirements for a scientific report, in particular if this report is founding policy making. Alternative methods for euthanasia of rodents have similar risks as gaseous anaesthetics produce the same behavioural effects to CO2 at induction concentrations and inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Costs will be higher than stated.



Worldwide Representative Replacement of the existing methods should be based on scientific evidence and evaluation of risks (and costs) of possible alternatives.



Worldwide yes As above, we reject the policy proposal on scientific grounds welfare effects are very questionnable and the costs would be high.

42



Worldwide yes As above, we reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk.




As above, we reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstract&list_uids=16542038&query_hl=3&itool=pubmed_docsum The second paragraph is not relevant.



Worldwide Yes I am not aware of scientific evidence demonstrating superior methods for humane euthanasia of large numbers of rodents. CO2 is acceptable if applied correctly. This should be taken care of by appropriate guidance/training.



National representative As above, I reject the policy proposal on scientific grounds: welfare effects are uncertain, alternatives are not present and costs are high. Alternatives are currently undefined and carry risk. http:// www.nature.com/nature/journal/v441/n7093/full/441570a.html




The evidence on the effects of CO2 on welfare are uncertain and hence we cannot support the analysis that there would be a great increase in animal welfare. However, we do support the view that there would be higher costs of alternative methods.

43



This is a sweeping statement – there is evidence to suggest that some anaesthetic gases are equally aversive as CO2. One must also note that there will be differences in the level of aversion experienced by different species. Further information can be extracted from the Newcastle Consensus Report (see below).


Public authority National Yes It is not common EU-policy to exclude a single agent at a point where disagreement on effect still exists.




See comment given above. CO2 is used to anesthesize pigs prior to slaugther! In Germany this roughly applies to 40 mio animals. The issue needs a much more careful cinsideration.

44






Public authority National Yes There is evidence that some of the adjuncts are also aversive. The same adjunct has not been shown to be appropriate for multiple species and therefore the potential costs are higher than would be predicted if the same adjunct could be used for all species. There is sufficient evidence in the literature that the use of the wrong adjunct can be aversive and therefore could be as detrimental to welfare as the use of CO2 alone.







yes It is not conclusively proven that the use of pre-anaesthetic use is a welfare advantage. Evidence is to be produced on NC3Rs website of research performed in this area. If given under appropriate conditions, CO2 may be no more stressful than pre-anaesthesia technique. Higher costs will occur using preanaesthetic.



45


Public authority National We are the National Competent Authority

The EFSA advice has not been peer reviewed and is flawed. There is evidence that some of the alternatives and supplements are also aversive.



see 1.4




Ban of CO2 is scientifically not justified! Especially for small rodents CO2 might be one of the least stressful method. If applied correctly, CO2 will quickly lead to unconsciousness (e.g. see Grahwit, 2005, "Euthanasia of small rodents - reasons, methods and assessment of animal welfare.", Dtsch. tierärztl. Wschr. 112: 95-97). Training on correct use of CO2 has likely a larger impact on animal welfare than the ban of CO2.




Public authority Worldwide yes I have experience with CO2 gas. Anethetizing rats and mice with CO2 gas and then killing them by decaptitation puts minimal stress or pain on them. It is important using CO2 gas not dry ice

46


Animal welfare must take precedence over euthanasia costs.




There is not yet a consensus on the welfare benefits of banning CO2 euthanasia.


Other European No See above X.e)




see 1.4


Other National Representative More research is required before deciding to ban use of CO2 euthanasia. See Newcastle consensus report at www.nc3rs.org.uk



47


Other Not applicable not applicable see above



Yes ther are higher costs but there is no consensus that other methods would result in significant improvements in animal welfare.




British Heart Foundation


National We are a medical research charity, of which there are many in the UK

No further comment



European Yes There is not yet a consensus about the welfare benefits of banning CO2 euthanasia

48

FELASA (Federation of European Laboratory Animal Science Associations)


European FELASA represents 13 European lab animal sci associations and therefore some 3-4000 members of those associations (scientists, veterinary surgeons, technicians etc.). See www.felasa.org

Very few research results demonstrate the aversiveness of CO2 and, in particular, the statement that CO2 is aversive to “all vertebrates” is an exaggeration. On the contrary, many other publications demonstrate its usefulness for euthanasia. Furthermore, recent studies by Flecknell et al have not been taken sufficiently into account in formulating this assessment.



National; European


The Physiological Society


Worldwide Many other Learned Societies aer also representative

The Technical Expert Working Group was unscientifically selective in its assessment of this topic. First there is new data, of which the TEWG was aware, that challenges the assumption of adverse welfare effects of CO2 euthanasia, and this data set needs to be incorporated before any final decision. Second, there needs to be a method superior to CO2 that is not only better for animals but also safe for operators. The former might exist, but the latter does not.

49

The Royal Society







The EFSA report is scientifically flawed and should not be used to set policy. A recent workshop in the UK has concluded that other gaseous anaesthetics produce similar behavioural effects to CO2. In addition to the unacceptable effects of the alternative gases they additionally carry significant cost to introduce and use. The evidence for the adverse effects has been mainly generated in rats whilst probably 80% of all rodents that are euthanased by this method are mice.





50



National YES Many publications demonstrate CO2 usefulness for euthanasia (Flecknell et al.) Further research is necessary before any clear recommendation can be made.




See below




See 1.4.



Worldwide No other organisations

We understand from the report of the Newcastle Consensus Meeting on Carbon Dioxide Euthanasia of Laboratory Animals (http://www.nc3rs.org.uk/downloaddoc.asp?id=416&page=292&skin=0) that whilst the use of CO2 is aversive to some species of animals; there is a need for further research before any type of prohibition is implemented, as there is insufficient information currently available on alternatives to CO2. Therefore, any proposed ban on the use of CO2 is premature.



Regional Yes see 1.4

51







Worldwide - -



National yes The statement of the Panel on Animal Health and Animal Welfare of the European Food Safety Agency about the CO2 method for rodents is scientifically inaccurate. A ban or recommendation of additional anaesthesia in combination with CO2 is unjustifiable. Other gaseous anaesthetics show similar behavioural effects at induction concentrations Inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. The inclusion of technical details in methodologies is not consistent with the purpose of a Directive and will limit the scope for future refinement of methodologies as new scientific data emerge. Additionally the costs will increase disproportionate.



Worldwide Representative As the literature on this topic is unclear, it is essential not to impose legislation that may prove unnecessary. The use of carbon dioxide administered appropriately by experts may prove to be less aversive than some of the literature suggests. More work is needed to assess this.



Worldwide representative CO2 is proven to be an anaesthetic in human if mixed with oxygen sufficient to avoid asphyxia

52


Breeder of animals




Regional; National


It is not conclusively proven that the use of pre-anaesthetic use is a welfare advantage. Evidence is to be produced on NC3Rs website of research performed in this area. If given under appropriate conditions, CO2 may be no more stressful than pre-anaesthesia technique. LASA working party (1997) and AVMA Panel (2000) reports on euthanasia indicate CO2 is satisfactory method & latter quotes work showing that rats in increasing concentrations of CO2 in home cage showed no behavioural or physiological signs of stress. Higher costs will occur using pre-anaesthetic. The same adjunct has not been shown to be appropriate for multiple species and therefore the potential costs are higher than would be predicated if the same adjunct could be used for all species. There is sufficient evidence in the literature that the use of the wrong adjunct can be aversive and therefore could be as detrimental to welfare as the use of CO2 alone.


National; European


Other methods for rodents are of similar level of risk, eg gaseous anaesthetics, and inert gases. The scientific literature does not cover rodents apart from rats. Hence the EFSA report lacks scientific basis and should not inform changes in the current directive. We recommend that the directive should not be prescriptive, to allow for the capacity to use improved methodologies and be informed by new data as these become available.


Regional; Worldwide

also others see 1.4

53


Local representative of national pharmacological research institutions

The use of CO2 gas for euthanizing small laboratory animals is a humane and pragmatic method.







The statement of the Panel on Animal Health and Animal Welfare of the European Food Safety Agency about the CO2 method for rodents is scientifically inaccurate. A ban or recommendation of additional anaesthesia in combination with CO2 is unjustifiable. Other gaseous anaesthetics show similar behavioural effects at induction concentrations Inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. The inclusion of technical details in methodologies is not consistent with the purpose of a Directive and will limit the scope for future refinement of methodologies as new scientific data emerge. Additionally the costs will increase disproportionate.

54

Question X.1.2.

Question text:

Statistics:




Comments received:





X.1.2a.

animal welfare: The use of anaesthetic gases such as halothane to render animals unconscious before applying CO2 greatly reduces the suffering of animals. In this respect the impacts to animal welfare would be high especially when considering the large numbers of animals this would concern. Do you support the preliminary findings? Yes/No/ No opinion

55




See comment on 1.1 above.





56





Ban of CO2 is scientifically not justified and has no animal welfare impact !

Hannu Komulainen


National; European

Is relevant It would be non-sense to use two gases for the same animal, the other for an unconscious animal!





Prof Alan Palmer



57





A undifferenciated prohibition of CO2 is scientifically not justified and has virtually no positive impact on animal welfare / suffering and other parameters.

Torgny Jeneskog, Umeå University, Sweden



Yes, guess so ... Please, see my response in 1.1a








XXX See 1.4




Yes Halothane can be adversive and it is not clear that suffering will be reduced. A careful cost-benefit must be established before a ban can be considered.




See 1.1

58










Halothane should certainly be avoided due to its toxicity of users. It is well known that nurses in operating rooms get health problems, unless very good systems are installed for removal of the halothane.




There is no sufficient scientific basis to abandon C02. Halothane, which is right now being abandoned for anaesthesia for toxucologic reasons in man is certainly not an alternative.



Local Yes During CO2 euthanisation there is no major suffering for the animals. The type of stress during primary halothane or primary CO2 exposure is likely to be the same.




Yes see 1.4

59




Yes Inappropriate use of halothane will also impact adversely on animals







The effect would be “+” at best.



National; Worldwide






See comments made in Aa introduction



Local yes See above X.e)

60



Local yes see Aa




Yes Not enough evidence to support this





The use of a volatile agent such as halothane is already been phased out of use.



Local Not applicable, I am not an organisation

Halothane is potentially human toxic (do your homework before making suggestions) and therefore its use must be balanced and used in appropriate conditions




No. The negative effects for animals of properly executed CO2 euthanasia as compared to other forms of euthanasia are still subject of debate.




61




no If CO2 is derived from gas tanks, and the cage of animals is slowly filled, these animals do not show any symptoms of stress or pain. If you place an animal directly into a container filled with anaesthetic gases, these animals show much higher stress symptoms than with slow increase of CO2 concentration.

Bayer HealthCare



1.2 There is no scientific consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems. http://by-ej-sid.bayer-ag.com/han/EJ-1648/www.nature.com/nature/journal/v441/n7093/full/441570a.html There is no data to confirm or refute if what is seen by people in some studies is seen in all rodent species. Even if this data did show a welfare issues, the concentration of halothane and other agents required for anaesthesia induction cause the same behaviours to a lesser or similar degree.



as above




There is no scientific consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems. There is no data to confirm or refute if what is seen by people in some studies is seen in all rodent species. Even if this data did show a welfare issues, the concentration of halothane and other agents required for anaesthesia induction cause the same behaviours to a lesser or similar degree. (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=retrieve&db=pubmed&list_uids=12120924&dopt=Abstract). Therefore if there is suffering, and this is disputed, it will be similar with halothane and CO2

62

sanofi pasteur R&D


Worldwide Vaccine field No if CO2 is well used



National yes There is no scientific necessity to do so, see also 1.1





Work done by Flecknell reveals that by using specially designed chambers that ensure unconsciousness before 50% concentration re not associated with pain activation. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=retrieve&db=pubmed&list_uids=12120924&dopt=Abstract) – show that Halothane may be associated with behaviours consistent with nociceptive system activation. If there is suffering and this can be discussed, the Halothane may be similar to CO2



Worldwide Yes The evidence to support this assessment is scant and contradictory.




no comment



Worldwide Yes More research is needed before the issue is regulated by a Directive




They are much more risk for human safety using halothane ! This gaz should be prohibited ! see above: no need of other gaz than O2 or air

63



National; Worldwide

yes See previous comment above. This statement cannot be accepted both for scientific, ethical and practical reasons.







See problem encountered with gaseous anesthesia.




There is no data available to determine what animals experience on a rising concentration of CO2 versus a rising concentration of halothane. Both agents are aversive though halothane may be less aversive. For this reason the preliminary finding is difficult to support as the benefit to animal welfare would not be high.



Worldwide No There is no great suffering when CO2 is used appropriately. Halothane is a safety hazard for humans. Some other volatile anaesthetics such as isoflurane have been considered by Flecknell and others to be at least as aversive as CO2 to rats at the concentration needed to anaesthetise them.



Worldwide pharmaceutical There is no scientific consensus that signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems.



Worldwide Yes In our experience, animals appear more distressed when exposed to a gaseous anaesthetic than to carbon dioxide at an appropriate flow rate. In addition, the use of anaesthetics to kill animals used for studies of the central nervous system is not possible due to the effects of the anaesthetic agent on brain biochemistry.

64




There is no scientific consensus that exposure to CO2 in a rising concentration below 50% represents a welfare problem.




Alternatives not available



Worldwide yes No scientific consensus that signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems (LINK). No data to confirm or refute if what is seen in some studies is seen in all rodent species. Even if data did show a welfare issue: concentration of halothane & other agents required for anaesthesia induction cause same or similar behaviours. (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=retrieve&db=pubmed&list_uids=12120924&dopt=Abstract). Therefore if there is suffering, & this is disputed, it will be similar with halothane & CO2








National Yes There is no clear cut scientific evidence that the signs seen in rats after exposure to CO2 represent significant welfare problems. There is no data to evaluate that what is experienced by human beings in some studies is seen in all rodent species. Halothane and other agents required for anaesthesia induction cause the same behaviours to a similar degree.

65



Worldwide Representative See the above comments (re: scientific evidence)



Worldwide yes There is no scientific consensus that exposure to CO2 in a rising concentration below 50% represents a welfare problem.



Worldwide yes There is no scientific consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems . There is no data to confirm or refute if what is seen by people in some studies is seen in all rodent species. Even if this data did show a welfare issues, the concentration of halothane and other agents required for anaesthesia induction cause the same behaviours to a lesser or similar degree.




The scientific discussion on the best method to use is an ongoing one. Whenever other new methods/ tools in any science area are developed, the scientific discussion between experts is restarted – and this for good reasons from the animal welfare point of view. Re CO2 use, there are still many different opinions of well known and accepted experts under discussion. This shows that no final decision on the CO2 use can be made for the time being. Thus an inclusion in the 86/609 seems to be premature.



Worldwide Yes Halothane is irritating, too.

66



National representative There is no scientific consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems. http:// www.nature.com/nature/journal/v441/n7093/full/441570a.html There is no data to confirm or refute if what is seen by people in some studies is seen in all rodent species. Even if this data did show a welfare issues, the concentration of halothane and other agents required for anaesthesia induction cause the same behaviours to a lesser or similar degree.




The evidence to support this assessment is scant and contradictory. Availability of halothane in the longterm is uncertain, as it is now rarely used in human medicine.



In February 2006, active researchers in this field met at Newcastle UK and have agreed a consensus report which will be available from www.nc3rs.org.uk. Within this report there are significant areas of disagreement with the EFSA opinion. It is therefore recommended that this issue, and any decision, should await publication of the consensus report and further consideration of the information contained therein.

67

Coenraad F.M.Hendriksen

Public authority National at the national level: yes

halothane is heptotoxic and for this reason of concern for the working environment (animal care staff)




see above comment






Public authority National Yes The current evidence for significant impact on animal welfare is limited to some species. Although the research indicates that adjuncts to CO2 may be of welfare benefit in these species, further work is required to allow an assessment of the impact of adjuncts to CO2





68



yes there is little evidence to support this contention, if CO2 is given in an appropriate manner





The evidence that the proper use of a rising concentration of CO2 produces unnecessary stress of suffering is not compelling – and there is some evidence that some inhalational anaesthetics inert gases may also be aversive. – thus it is not clear what the impact would be on animal welfare. The current evidence also suggests that no single adjunct would be appropriate for all species and stage of development, and the wrong adjunct could increase welfare costs. It may be some years before there is sufficient evidence to justify permitting/requiring or allowing the use of adjuncts and anything other than an option rather than the default method.



see 1.4

69




See 1.2; if applied correctly, CO2 is one of the least stressful method and will quickly lead to unconsciousness. Halothan is considered hepatotoxic! It will thus be of heath risk to the personal.




Public authority Worldwide yes Halothane exites the animals first. It is also toxic for the personal. Isoflurane is expensive




There has been a debate about the welfare aspect of CO2 euthanasia for some time, but there is not a clear consensus that any of the practical alternatives are more humane. Thus, it would be premature to legislate to ban it at this time.

70


Other European No See above X.e)




see 1.4


Other National Representative This is a sweeping statement – there is a need for more research on the animal welfare implications of use of CO2 in mice and of alternative methods of euthanasia (e.g. anaesthetic gases).



See above. In addition, the recommended method is inapplicable on large number of animals and may lead to everyone’s sleeping nearby!

71




Other Not applicable not applicable Again, I don't think this issue belongs here. More research is needed before having this isseu regulated by the Directive.







No further comment



European Yes There has been a debate about the welfare aspect of CO2 euthanasia for some time, but there does not appear to be a clear consensus that any of the practical alternatives are more humane. Thus, it would be premature to legislate to ban it at this time.

72




See comment on 1.1 above.



National; European





Comparison with techniques used by humans on humans suggests that CO2 is more humane than any used on humans.

73

The Royal Society







There is no scientific consensus that a rising concentration of CO2 produces suffering.



National yes There exists no scientific reason for a ban of CO2 euthanasia if done properly.

74




Despite its widespread use, CO2 euthanasia is not standardized (see Report of the ACLAM Task Force on Rodent Euthanasia, August 2005). This lack of standardization may be responsible for bad performance. The first step should be to set the technical requirements for this method of euthanasia. There is no worldwide scientific agreement regarding the aversive effect of CO2 and new data become regularly available (see JAALAS, 45, 4, July 2006, page 78, PS20). Halothane is a hazard for people. Only Isoflurane should be used and it is very expensive.







Worldwide - -



National yes Other gaseous anaesthetics show similar behavioural effects at induction concentrations. There is no consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems.




75



Worldwide yes From an human point of view the use of halothane should be avoided and other anaesthetic gases should be favourable in use (f. a. Isoflurane). If you use an anaesthetic gas for rendering the animals unconscious, you could use the same for euthanasia, which is more practicable, when looking at the gas piping through large facilities (you then need only one), using both would heavily increase costs for that piping Equipment. There are twi recommendations on Euthanasia in Lab. Animals one from the AVMA and one from the European panel which include the use of CO2. I am only aware of the study from the UK Specialist who indicates that the use could stress the animal. I think further studies are required which not only refer to the use of CO2, but to the comparism between Co2 and other euthanasia methods as well.



Worldwide representative having seen rodents killed with anaesthetic or CO2 + oxygen there is no difference


Breeder of animals



76


Regional; National


Justification: there is little evidence to support this contention, if CO2 is given in an appropriate manner. The current evidence for that exists for impact on animal welfare is limited to some species. Although the research indicates that adjuncts to CO2 may be of welfare benefit in these species, further work is required to allow an assessment of the impact of adjuncts to CO2


National; European


There is no clear agreement on the impact on welfare due to CO2 exposure. Further, the concentration of halothane and other anaesthetic agents have an equivalent effect.





Local representative Other gaseous anaesthetics show similar behavioural effects at induction concentrations. There is no consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems.




Other gaseous anaesthetics show similar behavioural effects at induction concentrations. There is no consensus that the signs seen in rats after exposure to CO2 in a rising concentration below 50% represent significant welfare problems.

77

Question X.1.3.

Question text:

Statistics:




Comments received:





X.1.3a.

Impacts to establishments: The use of anaesthetic gasses such as halothane or isoflurane can be effective but are more expensive than CO2 alone. One-time Investments between 13.000 Euro and 25.000 Euro depending on the configuration are required for a complete anaesthetic setup for rodents including an active scavenging system. Do you support the preliminary findings? Yes/No/ No opinion

78








XXX See 1.4




Halothane is a hazard for people. Only Isoflurane should be used and it is very expensive. One research institution does not mean only one setup to euthanize animals. In our institution with 10 animal facilities, 70 animal rooms and 14.000 mouse cages, there are as many as 20 setups for euthanasia with CO2 at different places. We could not afford to replace everyone of them with anaesthetic setups.



Local; National; European; Worldwide

representative of a research university

Many systems for delivering these anaesthetic gases are already in place and so such investments may not be needed.

79



Local Yes There is no advantage in using halothane or isofluorane instead of CO2 concerning animal welfare. Nothing is published on this matter in widely recognized scientific journals.



National; Worldwide


No, it is assumed that the cost of this proposal would simply be the one-off cost of buying the apparatus necessary for an alternative method in every establishment. However, all of the alternative methods of euthanasia require significantly more time and manpower for the same number of animals. This means that the proposal would result in substantially increased costs on an ongoing basis, without any certain animal welfare benefit.



Local yes No, there are not only higher costs for one-time investments but also for running the system, especially with isoflurane. Halothane is aversive and animals sacrificed with halothane can not be used for feeding reptiles etc.. Operational safety can be also a problem using halothane.



Local yes The costs in these relations should not be relevant if there would be an improvement for the animals. But there is no refinement for the animals.




No. The overall additional costs are far higher than the one-time investment for apparatus due to the lower efficiency of alternative procedures.




yes A great proportion of research establishments already have acess to the necesary equipment.

Bayer HealthCare



1.3 Costs include the unit costs of specialised anaesthetic equipment (€5,000 - €10,000) with one unit required for each area of the institution (there may be many), the costs of gas scavenging equipment (€5,000 - €10,000). Ongoing costs include the costs of the agents, maintenance costs for all equipment, greater staff time and training costs for a more complex to stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters.

80



As above, these gases are also aversive, but the degree is unclear, and costs increase because animals then contain xenobiotics after death. They cannot then be used for reptile and raptor feeding, and have to be disposed of as special waste. http://scienceandresearch.homeoffice.gov.uk/animal-research/publications/publications/guidance/breeding_supply.pdf?view=Binary




Costs include the unit costs of specialised anaesthetic equipment with one unit required for each discrete area of the institution (there may be many), the costs of gas scavenging equipment. Ongoing costs include the costs of the agents, maintenance costs, greater staff time and training costs for a more complex two stage process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. Halothane is increasingly going out of production. It has already been noted that halothane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed. The use of halothane will mean animals are regarded as clinical waste. They cannot be used for feeding raptors and reptiles. Therefore there will be significant clinical waste costs.



National yes Establishment of such equipment is a one-time cost. But consequential costs arise than, personal, more time, gases and so on. Without secured scientific background such emphasis should not be given. There are several reports about the negative effect of halothane on animals.



European Yes Although they are noticeably expensive, they are not required in such huge amounts as CO2. So the cost should be calculated accordingly.Additionally, animal resarch should be privilage and the institutions which have the qualifications should be allowed to do animal research. However, in today's regulations, even small sized labs can the this permit with a proper manipulation of the holes in regulation.

81





Volatile gas systems are expensive – in the region of €5000 to €10000. Each has to be connected to suitable scavenger system (depending on buildings – retrofit can be very expensive – in excess of 50 to 100,000). Each station needs calibration, scavenge packs need disposal. I have considerable experience with such system and I question the one off costs shown here. Finally animals that are euthanised by gases other than CO2 are a clinical hazard and cannot be used to feed raptors and reptiles. Clinical waste needs to incinerated which also adds to costs.




Actually yes and Halothane is a hazard for people. Only Isoflurane should be used and it is very expensive.




What are the basis for these costs ? A complete setup with exhaust system could be much more expensive, and compare to a CO2 euthanasia station, halothane and isoflurane is a risk for human safety.



National; Worldwide

yes See previous comment above. This statement cannot be accepted both for scientific, ethical and practical reasons. It is not a matter of cost. We could add some safety issues related to the control of gaseous anaesthetics but there is enough evidence if favour of a well conducted and well controlled CO2 euthanasia.



European Representitive This depends on the size of the establishment, and numbers of people that will need retraining. I expect it would cost E50,000 at my establishment, at least, and is more expensive to maintain, and administer. It is also more dangerous for the operators. I thought Halothane is Toxic in humans?




See above; ratio harm/benefit for CO2 is not clear




representative for MD&D

The costs are maybe underestimated. Moreover, animals having been anaesthesised with isoflurane are considered clinical waste (cannot be used for feeding raptors and reptiles) and costs for clinical waste incineration will also increase.

82




The figures cited would allow the procurement of the equipment but given the technical complexity of the architecture of an animal facility the costs above would not permit any building modifications which may be required to effectively scavenge the waste anaesthetic gases. It appears little regard has been paid to the implication of expanding the worker population exposed to halogenated gaseous agents. The devices for delivering and scavenging the agents must be regularly serviced and maintained as well as purchased. The environment must be monitored and there is a validation process to ensure the systems restrict the occupational exposure levels. Halothane will soon become unavailable meaning the more expensive alternative of isoflurane will be used. The exposure to rodents by halothane means that carcasses cannot be used for outlets such as food for birds of prey or snakes. The overall impact on welfare may be neutral or negative.



Worldwide No As well as the one-time investment, with which I agree, , there would be on-going costs of purchase of the anaesthetic, with little or no perceived benefit, and increased potential human health and safety concern.



Worldwide pharmaceutical The use of halothane is decreasing and isoflurane needs very expensive equipment (10.000 EUR for one isoflurane equipment versus 1500 EUR for an automated CO2 euthanasia equipment). There are also important safety issues, which are predominant with the use of isoflurane (exposure of <20ppm for the operators) which means that expensive scavenging equipment must be well designed and reliable.




We estimate costs of 15- 20K Euros per unit to provide anaesthetic equipment and scavenging (for human safety). A unit would be needed for each area where the work needs to be done.However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste, so that they may not be used for feeding raptors and reptiles. Therefore there will be significant clinical waste costs and increased incineration outflow.

83




Costs include the unit costs of specialised anaesthetic equipment (€8,000 - €10,000) for one unit, the cost sof gas scavenging equipment (€5,000 - €6,000). Ongoing costs include the costs of the agents, maintenance costs for all equipment, greater staff time and training costs for a more complex two stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. It has already been noted that halothane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste.



Worldwide yes Costs incl. unit costs (C) of specialised anaesth. equipment (€5K- €10K) with one unit required for each discretet area of institution, cost of gas scavenging equipment (€5K - €10K).Ongoing C include C of agents,mainetenance C for all equipment,greater staff time&training C for a more complex to stage halothane+CO2 process,disposal C for activated charcoal gas scavenging packs,or environmental C of venting ozone depleters. In addition halothane=increasingly going out of production.It has already been noted that halthane is aversive,other agents more so,&CO2 less so.However,significance of this aversion has not been assessed.Most significant:use of halothane will mean animals are regarded as clinical waste. They cannot be used for feeding raptors&reptiles.Therefore: significant clinical waste C,incineration outflow as more animals being bred,probable at lower standards of care outside research breeders.In summary:proposal will likely cost significantly more to make welfare worse!



National Yes Costs include the unit costs of specialised anaesthetic equipment (€5,000 - €10,000) with one unit required for each specific procedure room. It has also been noted that Halothane is aversive, but the significance of this has not been assessed.



Worldwide Representative For a medium-sized Facility, the present costs are absolutely underextimated, and do not consider all the indirect costs of maintenance.

84



Worldwide yes We estimate costs of 15- 20K Euros per unit to provide anaesthetic equipment and scavenging( for human safety). A unit would be needed for each area where the work needs to be done. Ongoing costs include the costs of the agents, maintenance costs for all equipment, greater staff time and training costs for a more complex two stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive (some other agents more so) and CO2 less so. However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste, so that they may not be used for feeding raptors and reptiles. Therefore there will be significant clinical waste costs and increased incineration outflow.



Worldwide yes Costs include the unit costs of specialised anaesthetic equipment (€5,000 - €10,000) with one unit required for each discretet area of the institution (there may be many), the cost sof gas scavenging equipment (€5,000 - €10,000). Ongoing costs include the costs of the agents, mainetenance costs for all equipment, greater staff time and training costs for a more complex tqo stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. In addition halothane is increasingly going out of production. It has already been noted that halthane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed.




Costs include the unit costs of specialised anaesthetic equipment (€5,000 - €10,000) with one unit required for each discretet area of the institution (there may be many), the cost sof gas scavenging equipment (€5,000 - €10,000). Ongoing costs include the costs of the agents, mainetenance costs for all equipment, greater staff time and training costs for a more complex tqo stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. In addition halothane is increasingly going out of production. It has already been noted that halthane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste. In summary this proposal will likely costs significantly more to make welfare worse!

85



Worldwide Yes My estimate of investment costs for a large establishment like a research center is higher than 25.000 EUR.



Worldwide Yes No it an extra cost burden being placed on the establishment.



National representative Costs include the unit costs of specialised anaesthetic equipment (€5,000 - €10,000) with one unit required for each area of the institution (there may be many), the cost sof gas scavenging equipment (€5,000 - €10,000). Ongoing costs include the costs of the agents, maintenance costs for all equipment, greater staff time and training costs for a more complex tqo stage halothane + CO2 process, disposal costs for activated charcoal gas scavenging packs, or environmental costs of venting ozone depleters. In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste. They cannot be used for feeding raptors and reptiles.

86



See entry under 1.2 above


Public authority National Yes Running costs will also be considerably higher than with CO2. Even with scavenging system occupational aspects are problematic.


Public authority National Yes The costs are not only in the set up of the equipment (set up estimate per system – likely to need 1 system per species if using different agents, since 1 agent is not appropriate for all species: 5100 euros for scavenger, basic system, cylinders, vaporiser and chamber: assume highest of estimates above may include piped gases). Ongoing monthly costs dependent on use level; of the order of 50 euros for cylinder rental, and between 0.45 to 2.10 euros per box fill (approx 20-30 mice or 5 to 10 rats) depending on agent used (halothane, iso or sevo), plus servicing costs.

87



yes Do you support the preliminary findings? No. These costs seem very high and lower costings seem likely in some circumstances. Where high throughput is required then this level of expense might be needed.There is no mention of the ongoing cost implications which would not be insignificant, given the probable non-availabilty of halothane due to reduced clinical usage because of human health concerns. The alternative agents are significantly more expensive



See above. There is information only in some contexts and for some species. Much further work is required to justify mandating the use of such adjuncts. Assuming that one system will be required per species (and perhaps also per stage of development), that a range of adjuncts will be required, and that there are ongoing additional operating and servicing costs, the cost estimate is too low.



see 1.4




There would also be a significant ongoing cost for the larger establishments as the alternatives to CO2 cannot handle as many animals, thus greater number of person-hours would be required.


Other European No No, there are not only higher costs for one-time investments but also for running the system, especially with isoflurane. Halothane is aversive and animals sacrificed with halothane can not be used for feeding reptiles etc.. Operational safety can be also a problem using halothane.


Other Not applicable not applicable cost calculation does not reflect ongoing costs an those for disposal. also, in most instances one unit is not suffieicent to serve the entire organisation.

88



The use of alternative agents to CO2 will have significant cost implications and there is no consensus that these costs would be justified by improvements in animal welfare.




No further comment

Eurogroup for Animal Welfare/EWLA


European Eurogroup/EWLA is made up of animal welfare Member Organisations and Observers in all Member States

It is not possible to have an opinion on the cost for reconfiguring anaesthetic setups without knowing how accurate the stated figures are and how they have been determined. In some instances it may be possible to modify existing equipment to incorporate the use of different or combinations of anaesthetics. In such instances the associated cost would inevitably be less than replacing an entire system Where there is a clear way of reducing suffering, this should be implemented regardless of cost.



European Yes There would also be a significant ongoing cost for the larger establishments as the alternatives to CO2 cannot handle as many animals, thus greater number of man-hours would be required.

89



National; European

Yes No, there are not only higher costs for one-time investments but also for running the system, especially with isoflurane. Halothane is aversive and animals sacrificed with halothane can not be used for feeding reptiles etc.. Operational safety can be also a problem using halothane.




Widespread use of inhalational anaesthetic agents is not to be recommended on human safety grounds; for instance low levels of halothane can have adverse effects on human foetuses, and widespread use therefore poses a risk that should not be condoned. In addition there would be very substantial additional costs that are not warranted by a marginal improvement of welfare over a period of a very few minutes of an animal’s lifespan. It is notable that in humans, hypercapnia causes confusion and even euphoria before loss of consciousness.

The Royal Society




90




There is no scientific consensus that a rising concentration of CO2 produces suffering.




Halothane is a hazard for people. Only Isoflurane should be used and it is very expensive.







National yes Till now other methods for killing animals are not scientifically tested. The impact of using additional anaesthetic methods in combination with CO2 for animal welfare purposes is unclear. It does not make sense to calculate costs for an unproven method. In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive.




91


Regional; National


These costs seem very high and lower costings seem likely in some circumstances (my estimates of set up costs are closer to 5100 euros for a basic system using gas cylinders, but likely to need 1 system per species if using different agents, since 1 agent is not appropriate for all species). Where high throughput is required then the level of expense quoted in the document might be needed. The costs are not only in the set up of the equipment. Ongoing monthly costs are not mentioned but will also occur, which include the cost of the agent, scavenging and servicing and which would not be insignificant. There is also the probable non-availability of halothane due to reduced clinical usage because of human health concerns. The alternative agents to halothane are significantly more expensive and their usefulness as pre-stunning agents may not been tested widely or at all in some cases.


National; European


There will also be ongoing costs of consumables, equipment and staffing, with an impact on disposal costs and on environmental ozone.


Local representative Till now other methods for killing animals are not scientifically tested. The impact of using additional anaesthetic methods in combination with CO2 for animal welfare purposes is unclear. It does not make sense to calculate costs for an unproven method. In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive.




Till now other methods for killing animals are not scientifically tested. The impact of using additional anaesthetic methods in combination with CO2 for animal welfare purposes is unclear. It does not make sense to calculate costs for an unproven method. In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive.


National Not sure Gases such as argon are supposed to be better than CO2 because animals have not evolved specific receptors to detect it. Oxygen depletion is also not supposed to be as aversive. Sorry I have no references for this – I have seen the data presented as talks at conferences. These methods would not require halothane or isoflurane. Would they be less expensive?

92

Question X.1.4.

Question text:Justification (open text box)

Statistics:(no numbers registered as this was not a closed question)

Comments received:





X.1.4. Justification

Academical Medical Centre Amsterdam


Worldwide Yes it is representative

Animal welfare and public relations are more important than economical costs.

C. Norman Scholfield



yes CO2 not humane but cervicla dislocation or blow to head is Anaesthetics also involve an unpleasant excitment stage

93




Further research is necessary, perhaps funded under FP7, before any clear recommendation can be made on the use of CO2 for euthanasia,.

Elwin Rombouts User of animals -public sector

National; Worldwide

More relavant organisations, University, Biomedical Research

To my experience mixing CO2 with oxygen (thus keeping CO2 levels relatively low) reduces aggressiveness, at least rodents do not show any signs of severe irritation before they are rendered unconscious. This would mean less of an investment. Furthermore use of anaesthetic gasses needs more experience of users and higher maintenance in order to prevent exposure of workers to high levels of these gasses.

Hannu Komulainen


National; European

Is relevant Has anybody thought, how large problem the use of anethetic gases cause to health of the workers. They certainly will be regularly exposed. Halothane is a liver toxic gas. CO2 is inert in that respect. This point should be taken seriously. Could be an option but not a requirement.

Prof Alan Palmer



94







Local Yes 1.3. Running costs will also be considerably higher than with CO2. Even with scavenging system occupational aspects are problematic.




XXX The Technical Expert Working Group was unscientifically selective in its assessment of this topic. First there is new data, of which the TEWG was aware, that challenges the assumption of adverse welfare effects of CO2 euthanasia, and this data set needs to be incorporated before any final decision. Second, there needs to be a method superior to CO2 that is not only better for animals but also safe for operators. The former might exist, but the latter does not. Widespread use of inhalational anaesthetic agents is not to be recommended on human safety grounds; for instance low levels of halothane can have adverse effects on human foetuses, and widespread use therefore poses a risk that should not be condoned. In addition there would be very substantial additional costs that are not warranted by a marginal improvement of welfare over a period of a very few minutes of an animal’s lifespan.



Local; National; European; Worldwide

representative of a research university

Research needs to be carried out as a matter of urgency to develop a replacement for CO2. There is enough evidence to show its aversiveness and that it causes poor welfare in all species tested. What is needed are replacement non-aversive methods as are being developed for the farmed species. There are several approaches being tried, not just alternative gases, and many of these newer methods could be used with the same infrastructure and equipment as CO2. The cost of using an alternative gas such as argon would be minimal compared with the overall real costs and investment in each animal, and alternative costs are likely to be heavily influenced by what alternative method is being used. 1.1 I do agree with this statement but unless alternatives were developed. Some opportunities are available. The higher costs are relatively trivial in comparison with the overall investment in an experimental animal, probably less that 0.01% of the real costs involved.

95




Better teach investigators how to use CO2 euthanasia than looking for alternatives




A technical detail should not become part of a directive. It is also not possible to deal with such details: the issues are not yet entirely settled. Final judgement would have to deal with concentrations, flooding, prefilling, potential species differences etc. There is no sufficient scientific basis to abandon C02 now, and costs for replacement would be high.





The scientific literature on this topic is far from being clear. It is essential that more research is devoted to the topic of euthanasia of laboratory animals BEFORE legislation is imposed. As the literature on this topic is unclear, it is essential not to impose legislation that may prove unnecessary. The use of carbon dioxide administered appropriately by experts may prove to be less aversive than some of the literature suggests. More work is needed to assess this



National yes, competitive in the field of small animal research

I would plea for additional methods. Animals can be drugged by food (for instances benzodiazepines mixed in jellies or other things these animals like). When CO2 is added on top of these CNS depressing drugs less harm may be done. Clearly more research is needed. Costs for gasses (halothane and isoflurane) are too high.

96




Yes The use of carbon dioxide gas for anaesthetizing animals combined with rapid decapitation is a humane and pragmatic method for euthanasia of laboratory animals. Furthermore, the sequential combination of a volatile anaesthetic with cabon dioxide does not make sense. Why not simply use an overdose of the anaesthetic? As the literature on this topic is unclear, it is essential not to impose legislation that may prove unnecessary. The use of carbon dioxide administered appropriately by experts may prove to be less aversive than some of the literature suggests. More work is needed to assess this.




The health and safety impact of using halothane in particular, but also isoflurane for ALL C2 euthanasia is HIGH. Staff will be exposed to these gases at a much greater extent than now. Halothane is hepatotoxic and chronic exposure to this gas is undesirable. We also need to consider that establishments will then need to stock large numbers of bottles of these potent agents for this purpose, which will inevitably involve misuse and accidents.



Regional yes The scientific literature on this topic is far from being clear. It is essential that more research is devoted to the topic of euthanasia of laboratory animals BEFORE legislation is imposed.




Yes More research required.




No A report from a recent workshop on CO2 euthanasia, soon to be published by the UK National Centre for the 3Rs, concludes that it is premature to ban it as there is no practical alternative that could be said to be more humane. In addition, it is assumed that the cost of this proposal would simply be the one-off cost of buying the apparatus necessary for an alternative method in every establishment. However, all of the alternative methods of euthanasia require significantly more time and manpower for the same number of animals. This means that the proposal would result in substantially increased costs on an ongoing basis, without any certain animal welfare benefit.

97




Yes More work is needed before the ban on Co2 euthanasia is decided.




Yes Costs of anaesthetic agents are extremely high. This has all got to be evidence based, not anecdotal



National; Worldwide

yes, major UK research organisation for aquatic organisms and environment

Without a technical briefing it is not possible to cost the installation of a new system, but we would certainly comply with any regulatory change. Another implication may be staff-training, if anaesthesia is regulated as a veterinary rather than an experimental procedure.





Our establishment has already moved to the use of isoflurane rather than halothane.



Local Not applicable, I am not an organisation

One should provide a range of euthanasia techniques that are appropricate to the species and stage of development (e.g as is already done in the UK)

98



Not applicable No I think that CO2 should be banned. The use of halothane or isoflurane it is not more expensive than implementing all the housing, personnel and other changes, do not see why should it be not implemented as well.




no Detailed description of a suitable method to kill rodents with CO2 can be obtained from the AVMA at oacu.od.nih.gov/ARAC/EuthCO2.pdf or the NIH



Worldwide No there are other similar organisations

Halothane is toxic and substitutes may be little better. However, some agent to be used with carbon dioxide is necessary. Many establishments already use gaseous anaesthesia so costs should be low.




It is a University, the question is meaningless

I strongly support the view that the use of CO2 in conscious animals should be prohibited. From both simple observation of the procedure in rodents and human experiments involving rebreathing without CO2 scrubber, it is clear that the procedure is extremely unpleasant for the subject. The added "cost" to most institutions would be negligible, since they are likely to have the relevant equipment already.

Bayer HealthCare



In addition halothane is increasingly going out of production. It has already been noted that halothane is aversive, other agents more so, and CO2 less so. However, the significance of this aversion has not been assessed. Most significant of all is that use of halothane will mean animals are regarded as clinical waste. They cannot be used for feeding raptors and reptiles. Therefore there will be significant clinical waste costs, incineration outflow as more animals being bred, probable at lower standards of care outside research breeders. In summary this proposal will likely costs significantly more to make welfare worse! The Nature paper just give hints that rising percentage of CO2 might be unethical to use, but even this is disputed. Putting the animals directly into high CO2 concentrations seems to overcome the issue.

99



The EFSA report is scientifically inaccurate as above. Alternative methods for rodents carry similar or greater risks: gaseous anaesthetics produce similar behavioural effects to CO2 at induction concentrations, inert gases such as argon lead to adverse clinical signs (eg. Most recently; Behavior, Heart Rate, and Activity Levels Used as Indicators of Stress in Rats during Argon or Carbon Dioxide Euthanasia. TH Burkholder et al Journal of the American Association for Laboratory Animal Science Vol 45, No 4 P 78 July 2006) and nitrogen is ineffective. Scientific literature on current CO2 issues is on rats, not mice and other rodents. This inclusion of detail in a Directive and will limit the possibilities for the future refinement of methodologies as new scientific data emerge. Financial costs are higher than listed, for equipment, safety, and wastage of animals that can be used for animal feed.




Halothane is well known to carry a risk of hepatotoxicity in humans, so staff health and safety could be compromised through its regular use.





This proposition is based on misleading and incorrect data. See current work done by Flecknell for newest data.



Worldwide Yes Alternatives to CO2 would certainly require investment in more complex equipment. Control of exposure of staff is very important with some of the volatile anaesthetic alternatives. Several reviews and guidelines have been published where CO2 is an acceptable euthanasia method including the Euthanasia of Experimental Animals Report (XI/738/EEG). Laboratory Animals (1996) 30, 293-316 and (1997) 32, 1-32, JAVMA (2001) 218, 669-696.

100




- The list of human method of killing should not be included in the Directive , publications and reports of experts are available and new methods can occur before revision of the directive - Technical and scientific data on CO2 in rodents must be take into account and practical use (Good practice for use of CO2) must be defined (different advices in the literature) Halothane is hazardous



Worldwide Yes The additional use of fluorinated carbohydrates (halothane) will cause air pollution. We support EU-funded studies to find real alternative methods and to test the effects of halothane and CO2 in various species at different ages.







no comment



Worldwide There are other relevant organisations

Halothane and isoflurane are chlorofluorocarbon molecules. The potential risk for the environment should be taken into account when proposing new methods to be used on a large scale.



Worldwide No The information on CO2 presented by the EFSA working group has mixed up results from multiple species, to come up with an erroneous conclusion that CO2 should not be used in rodents. Scientific-based work in rats has indicated no such welfare concerns if CO2 is used appropriately. This is in line with my personal view having witnessed its use in many cases over many years. Effort should be made to educate people about the correct way to use CO2, ie by gradual exposure rather than 100% exposure. No credible, safe and economic alternatives currently exist.

101



Worldwide pharmaceutical Guidelines on particular methods of euthanasia should not be included in a Directive as these must be able to change easily following advances in scientific knowledge. On the other hand, encouragement for further research and advise on alternatives where concerns arise, should be seeked.



Worldwide yes -





No further comment



National Yes The report provided by EFSA does not meet the standards and requirements for a scientific report, in particular if this report is founding policy making. Alternative methods for euthanasia of rodents have similar risks as gaseous anaesthetics produce the same behavioural effects to CO2 at induction concentrations and inert gases such as argon lead to clinical signs that require investigation and nitrogen has been shown to be ineffective. Costs will be higher than stated. There is no clear cut scientific evidence that the signs seen in rats after exposure to CO2 represent significant welfare problems. There is no data to evaluate that what is experienced by human beings in some studies is seen in all rodent species. Halothane and other agents required for anaesthesia induction cause the same behaviours to a similar degree.

102



Worldwide yes A meeting was held earlier this year in Newcastle, UK where experts discussed the scientific evidence about the use of CO2. the outcome of this meeting was not in agreement with the findings of the EFSA report. The report of the meeting will be published in August 2006 at http://www.nc3rs.org.uk/ It is important that this is considered before any new regulation is drafted. We believe that the Directive should not specify euthanasia methods in detail. This would hinder the adoption of new methods which will be developed with the advance of scientific knowledge and technology. Methods use should be those of current best practice and could be determined by the ethical review process,




Maintenance costs should not be forgotten, as well as training of staff on how to do the new methods. However, these additional costs would not be a barrier for our company – as soon as the new methods are scientifically proved as being better for the animals.



National representative Therefore there will be significant clinical waste costs, incineration outflow as more animals being bred, probable at lower standards of care outside research breeders. In summary this proposal will likely costs significantly more to make welfare worse! The Nature paper just give hints that rising percentage of CO2 might be unethical to use, but even this is disputed. Putting the animals directly into high CO2 concentrations seems to overcome the issue.




Alternatives to CO2 would certainly require investment in more complex equipment. Control of exposure of staff is very important with some of the volatile anaesthetic alternatives.

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The APC does not support regulating euthanasia methods at this level of detail. With regard to 1.2, the use of anaesthetic gases may also compromise the work being carried out such as when the experiment being conducted involves collection and analysis/use of tissues.




The conclusion reached by a recent UK National Centre for the 3Rs conference in Newcastle, was that more research is required before deciding whether to ban the use of CO2.



yes This is not the time to be making this decision. Much data is being acquired in this field. It is not conclusive.

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Public authority National Yes You must be allowed to use CO2 alone. There are some - but not overwhelming, and also contradictory - data to support an adversiveness for CO2-inhalation in some species. BUT the adversiveness of a certain agent/method has to be put in relation to the adversiveness of other possible agents/methods. For many (most?) of those, including "anaesthetic gases", not much data are available to support their supremecy over CO2. Physical methods and injection techniques (overdose of anaesthetic) are indeed "stressful" per se, to the extent that the adversiveness of CO2-inhalation might in fact be negligable. [This might be arguments in general for more scientifically based studies on euthanasia methods!] -



Yes The use of halothane should be strictly forbidden because of its hepatic toxicity on mammals, especially humans.



The basis of the costing is not provided. The analysis above is silent on potential staff health, environmental and health and safety issues. Given the current state of knowledge it is not possible to construct a defensible case for moving from current to novel systems.


Public authority National yes Possible health risk for personel should be considered.



This issue has yet to be resolved scientifically. There is evidence that other inhalational agents may be aversive and there may be species differences in aversiveness. Currently there is insufficient scientific evidence to support this proposal. Also need to consider the acceptance of use of other agents for some studies, particularly regulatory studies where results may be influenced by interference of sedative or anaesthetic agents. Estimates of cost of implementation seem inaccurate and ongoing running costs are not considered and could be considerable. Overall this proposal is very detailed for a general directive. Changes in the scientific understanding and potential future developments may render this detail out of date rapidly.

105


Euthanasia means the humane destruction of an animal accomplished by a method that produces rapid unconsciousness and subsequent death without evidence of pain or distress, or a method that utilises anaesthesia produced by an agent that causes painless loss of consciousness and subsequent death The use of CO2 alone is unacceptable - whether administered under ‘optimal conditions’ or not (see: www.aphis.usda.gov/ac/cfr/9cfr1.html).




The Technical Expert Working Group was unscientifically selective in its assessment of this topic. First there is new data that challenges the assumption of adverse welfare effects of CO2 euthanasia, and this data needs to be assessed before any final decision is made. Second, there needs to be a method superior to CO2 that is not only better for animals but also safe for operators.

Cris Iles-Wright Other Not applicable Not applicable CO2 is routinely administered incorrectly and equipment is routinely insufficient for requirements outlined by legislation. Whilst considering killing methods it is also important to consider why these animal are bred in such large numbers, especially considering their irrelevance for medical requirements.

INRA center of Jouy-en-Josas Working group in animal experimentation ethics

Other Local it is representative at the local level

According to our animal technicians and our observations, we noticed that the use of anaesthetic gas before applying CO2 would avoid the period (around 10-15 seconds) during wich rodents suffocate because of lack of O2 replaced by CO2 until saturation. The anaesthetic gases can be used at room temperature avoiding the thermal stress when using CO2 at very low temperature when it arrives in the euthanasia cage. It would avoid also auditive stress due to the arrival of CO2 under high pressure. It has been experimented that reduced pessure in CO2 arrival decreases the rodents stress (excitation). For these 3 reasons, we think the use of isoflurane before applying CO2 in the same cage, instead of halothane (wich is prejudicial for humans), would increase animal welfare. it would cost more than 10 000 euros to add the system of anaesthetic gaz arrival in CO2 cages .




According to our experience, there is no scientific reason for a ban of CO2 euthanasia if done properly.

106

Royal Netherlands Akademie of Arts and Sciences

Other National Yes, representative of the scientific community

Comment to 1.2: Due to occupational safety hazards it is not advisable to work with halothane. We agree that animal welfare increases when an animal is rendered unconscious before exposing it to 100% CO2. There are however different ways to expose animals to CO2. In the Netherlands e.g. its very common to use 30-40% O2 in combination with 60-70% CO2 to render an animal unconscious without excitation before exposing it to 100% CO2.



1.3. The impact can be even worse (- -): costs may be higher because of the occupational risk in using volatile anaesthetics in large volume and for long exposure.


Other European ECPA represents the manufacturers of crop protection products (pesticides)

Question 1.2: Regarding CO2, when correctly used a rising CO2 level should be applied as this reduces animal stress rendering rapid unconsciousness and death. When used in this way anaesthetic gases may unnecessary.


Other National representative There has been considerable research into the use of CO2 for euthanasia in recent years. Many of the identified problems can be prevented by careful use of the agent. Rendering the animals unconscious with another gaseous agent prior to the introduction of carbon dioxide would also prevent these problems, but adds to the cost and also to the complexity of the equipment required, since rather than a single gas cylinder and induction chamber, two gas cylinders and a vaporiser would be needed. Until a cheap and reliable alternative without the welfare problems has been identified, the impact of removing CO2 as an agent for euthanasia would be considerable.

107


Other National There are other similar organisations

There seems not yet to be enough scientific evidence to ban the use of CO2. Halothane, being toxic to the personnel, should not be recommended as the basic anesthetic before the euthanasia. This is the area were more research is needed, especially to ensure that for example the tissues collected would not have been effected by euthanasia agent. If scientific evidence shows that there is a welfare need for anesthesia before CO2, the method of euthanasia should be changed after a transition period of some years. This is an important issue and financies should not be the main point in making decisions.



National Yes Ban of CO2 is scientifically not justified at this time. Further experimentation including direct comparisons with anesthetic gases in the same experimental models is necessary to identify the quickest and least aversive method for euthanasia.




The proposal to ban CO2 euthanasia is premature as there is no practical alternative that has been shown to be more humane.

Eurogroup for Animal Welfare/EWLA


European Eurogroup/EWLA is made up of animal welfare Member Organisations and Observers in all Member States

Although CO2 euthanasia is regarded as humane and acceptable by some scientists, recent reviews have identified serious welfare concerns associated with the technique (Conlee et al. 2005, EFSA 2005). A meeting of experts in the field, held in 2006, arrived at a consensus view that there is no “ideal” way to euthanase animals using CO2 (Hawkins et al. 2006). Exposure to high concentrations causes pain, and exposure to rising concentrations may cause other adverse effects such as dyspnoea, which can be highly distressing. The scientific literature regarding the effects of CO2 inhalation is far from complete and leading researchers in the field have recommended that further studies, taking a multidisciplinary approach, are required (Hawkins et al. 2006). In view of this, animals should not be exposed to carbon dioxide unless they have been rendered unconscious first, using an appropriate non-or minimally aversive gaseous anaesthetic agent (see Leach et al. 2003).

108

European Coalition to End Animal Experiments (ECEAE)


Worldwide Representative (the only EU coalition focusing on animal experiments)

We support these proposals on the grounds of increased animal welfare. It is important that the use of anaesthetic gases are fully substituted for this form of euthanasia and we are glad that the SCAHAW report has been mentioned. We would point out however that there are additional positive impacts for this option; public perception of the industry, wider societal impacts, positive impacts on researchers and technicians. Even in the absence of consideration of these it should be noted that the +++ for animal welfare minus the one – for cost should result in an overall positive impact of ++.




Further research is necessary, perhaps funded under FP7, before any clear recommendation can be made on the use of CO2 for euthanasia,.

109

PETA Europe Ltd.


Worldwide both The use of CO2 as a means of euthanasia is both painful and inhumane. Research has shown that death by CO2 results in the swelling and hemorrhaging of the lungs (Danneman et al. 1997). Animals have been noted as reacting with a panic – type response to being trapped in enclosures with elevated CO2 levels (Conlee et al. 2005).The EFSA Panel’s findings and recommendations (X.d above) must be taken into account in order to benefit animal welfare. The above-mentioned negative impact is financial, but it is inappropriate to justify animal suffering on the grounds that it will save money. The animal research industry is highly profitable, and the additional cost of anesthetic gasses is one that the industry should embrace willingly in order to demonstrate its commitment to animal welfare.

PROFESSOR RD COMBES - FRAME


Worldwide THERE ARE OTHERS

There are uncertainties relating to the humaneness of some techniques, including the use of carbon dioxide. Although many laboratory animals, especially rodents, are killed using CO2 according to a variety of protocols, there is currently no definitive guidance on whether and how CO2 can be administered humanely. There is also uncertainty about the feasibility of using alternative gaseous euthanasia agents, with respect to both animal welfare and human health and safety. A recent concensus meeting on Carbon Dioxide Euthanasia of Laboratory Animals (Newcastle, February, 20060 concluded that more research is needed into the physiological and responses to a range of gaseous agents, to identify possible alternatives to CO2 and define good practice for killing with carbon dioxide.

110

The National Anti-Vivisection Society



The NAVS is the leading and oldest anti-vivisection organisation in the world. There are a number of other relevant anti-vivisection organisations around the world with expertise in this field.

Problems associated with use of CO2 are likely to have a severe negative welfare effect, such as epistaxis and damage to nasal mucosa CO2 can be aversive to breathe, and animals may avoid the gas and either remain alive or suddenly get exposed to high concentrations whilst conscious (1). NAVS investigations have revealed that the CO2 method causes stress as rodents were observed frantically climbing over one another, trying to burrow into corners and trying to escape from the chamber. Gassing equipment was often faulty and measures to ensure and confirm death often not taken (2). If CO2 is to be used, it is imperative that stringent operating procedures be followed, but our investigations have shown time and time again that this is rarely the case. We suggest other methods, such as a mixture of gasses to be more humane. 1 Wolfensohn & Lloyd 2003. Handbook of lab. Animal Man. & welfare. Blackwell 2 Access denied 1996. NAVS report http://www.navs.org.uk/publications/reports/c=1

The Royal Society



We believe that the AHWP report to the EFSA failed to adequately review the scientific data on this issue. Furthermore, we understand from research published on the website of the UK’s NC3Rs (http://www.nc3rs.org.uk/downloaddoc.asp?id=416&page=292&skin=0) that whilst the use of CO2 is aversive to some species of animals, there is a need for further research before any type of prohibition on the use of CO2 is implemented. There is also uncertainty about the feasibility of using alternative gaseous euthanasia agents, with respect to both animal welfare and human health and safety. For these reasons we consider that a proposed prohibition on the use of CO2 is premature.

111

Universities Federation for Animal Welfare


Worldwide There are others There is good evidence that CO2 is highly aversive to rats above about 16%, whilst approx 30% is required to achieve loss of consciousness. CO2 appears to be less aversive to mice. A group of experts recently met at Newcastle and are producing a consensus report. Any decision should wait until this report is published.



European FEPS is an umbrella organization for national Physiological societies in Europe. See further http://www.feps.org

I (Hultborn) have personally participated on a panel to judge CO2 anesthesia several years ago (Denmark – slaughtering of pigs) – I do not really recognize/ remember the conclusions referred to here. I would also like to refer to the comments made both by EBRA (see EBRA Bulletin June 2006; European Biomedical Research Association) and the Royal Society on this point.




both please refer to statements by ECEAE and PETA



Worldwide Yes Recent evidence regarding concerns with carbon dioxide euthanasia justify a ban its use as a sole euthanasia agent, regardless of the financial costs to implement a system to render the animals unconscious first with an anaesthetic gas. Cost concerns should not override animal welfare concerns not only because of the welfare of the animals, but because public support for animal research drastically declines when the animals experience pain and distress. A review of carbon dioxide euthanasia by Conlee, et al (2005) Laboratory Animals, 39, 137-161 includes numerous arguments against the use of carbon dioxide as a sole euthanasia agents as well as recommendations regarding other methods, including the use of anaesthetic gases in combination with carbon dioxide.

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The use of halothane requires extremely expensive volatization and scavenging equipment. It is unlikely that animal facilities will be able to afford the investment. Continued use of isoflurane is prohibitively expensive. If CO2 is prohibited, it is likely that some animal care facilities will insist that researchers sacrifice animals with halothane but will not provide the necessary equipment. Continued exposure to halothane can cause life-threatening liver damage. Therefore much more work is needed to find a better alternative to halothane (or cheaper than isoflurane) before C02 is prohibited




A more thorough investigation of the aversiveness of CO2 needs to be conducted. In practce, particularly in small animals, it may be the most human method available.



Regional Yes The scientific literature on this topic is far from being clear. It is essential that more research is devoted to the topic of euthanasia of laboratory animals BEFORE legislation is imposed.




One other One can not understand that this questionnaire is limited to the fate a laboratory animals only when no mention is made of methods of substitution more trustworthy such as toxicogenomics on human cells.



Worldwide - -

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Worldwide yes From an human point of view the use of halothane should be avoided and other anaesthetic gases should be favourable in use (f. a. Isoflurane). If you use an anaesthetic gas for rendering the animals unconscious, you could use the same for euthanasia, which is more practicable, when looking at the gas piping through large facilities (you then need only one), using both would heavily increase costs for that piping Equipment. There are twi recommendations on Euthanasia in Lab. Animals one from the AVMA and one from the European panel which include the use of CO2. I am only aware of the study from the UK Specialist who indicates that the use could stress the animal. I think further studies are required which not only refer to the use of CO2, but to the comparism between Co2 and other euthanasia methods as well.


Breeder of animals




Regional; Worldwide

also others The scientific literature on this topic is far from being clear. It is essential that more research is devoted to the topic of euthanasia of laboratory animals BEFORE legislation is imposed. Moreoever, the sequential combination of a volatile anaesthetic with CO2 does not make sense. Why not simply use an overdose of the anaesthetic?

Documents

Excel Sheet semifinal CO2 rev1 - European Commissionec.europa.eu/environment/chemicals/lab_animals/pdf/x_co2.pdf · stress and subsequent stop of oxygen supply causes final euthanasia