Alternative methods Alternative methods contemporary and future situationcontemporary and future situation

in the Czech Republicin the Czech Republic

Dagmar Jírová,M.D., Ph.D.Dagmar Jírová,M.D., Ph.D.Department of ToxicologyDepartment of ToxicologyNational Institute of Public HealthNational Institute of Public HealthPraha, CRPraha, CR

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Protection of Animals Used for Experimental Purposes legislation in the CR

Directive 86/609/EEC,Directive 86/609/EEC, on protection of animals used for on protection of animals used for experimental purposesexperimental purposes

is implemented inis implemented in

Act No.246/1992 Sb. , Act No.246/1992 Sb. , on the protection of animals against crueltyon the protection of animals against cruelty

Decree No. 207 / 2004 / Coll., Decree No. 207 / 2004 / Coll., on protection, breeding and use of on protection, breeding and use of animals for experimental purposesanimals for experimental purposes

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Act No.246/1992 Sb. ,Act No.246/1992 Sb. , on the protection of animals against crueltyon the protection of animals against cruelty

Regulating :Regulating : definition definition of crueltyof cruelty, , possible purposes / procedures to kill animalspossible purposes / procedures to kill animals protection of animals during protection of animals during transporttransport protection of protection of farmfarm animals animals protection of protection of domestic animalsdomestic animals / breeding of special interest / breeding of special interest protection of protection of wildlife specieswildlife species of animals of animals protection of animals used for protection of animals used for experimental purposesexperimental purposes : : article 15 - 18article 15 - 18 competent authorities competent authorities responsible for protection of animals : responsible for protection of animals :

Ministry of Agriculture, Central Commission for Animal Welfare (consultant Ministry of Agriculture, Central Commission for Animal Welfare (consultant body to the Minister), State Veterinary Administration, Municipal Authoritiesbody to the Minister), State Veterinary Administration, Municipal Authorities

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Act No.246/1992 Sb. ,Act No.246/1992 Sb. , article 15 - 18article 15 - 18

animals used for experimental purposes animals used for experimental purposes

Possible Possible purposespurposes to conduct experimentsto conduct experiments on animals, including : on animals, including : - ban on testing of cosmetics and their ingredients, ..... weapons- ban on testing of cosmetics and their ingredients, ..... weapons - ban on use of abandoned animals , .... animals of endangered species- ban on use of abandoned animals , .... animals of endangered species - priority to use animals bred for experimental purposes- priority to use animals bred for experimental purposes

Experiment can only be performed, if Experiment can only be performed, if provedproved that no alternative methods are that no alternative methods are availableavailable ; ; unavoidable pain and distress should be unavoidable pain and distress should be minimized.minimized.

ObligationObligation to prepare to prepare project of the experimentproject of the experiment , it has to be, it has to be approved approved by by Animal Wefare Commission of the respective MinistryAnimal Wefare Commission of the respective Ministry

Requirements on Requirements on qualification of personnelqualification of personnel conducting experimentsconducting experiments RulesRules on breeding, care, accomodation and transport on breeding, care, accomodation and transport of experimental of experimental

animals, on animals, on accreditationaccreditation of breeders, suppliers and users, on of breeders, suppliers and users, on animal animal welfare committeeswelfare committees of these facilities, of these facilities, reports reports on number and species used.on number and species used.

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Decree No. 207 / 2004 Coll.,Decree No. 207 / 2004 Coll., on protection, breeding and use of animals for on protection, breeding and use of animals for

experimental purposesexperimental purposes

25 mandatory items to be addressed in project of the experiment25 mandatory items to be addressed in project of the experiment, including , including reasoning for unavoidability of the experiment, absence of alternatives, reasoning for unavoidability of the experiment, absence of alternatives, justification of species selection, number of animals, possible anesthesia, justification of species selection, number of animals, possible anesthesia, qualification of personnel, laboratory eqiupment .....qualification of personnel, laboratory eqiupment .....

Requirements on application for accreditationRequirements on application for accreditation of the facility. of the facility.Requirements on qualification of personnel Requirements on qualification of personnel conducting experiments : obligatory conducting experiments : obligatory

examination in elementary anatomy and physiology of laboratory animals, examination in elementary anatomy and physiology of laboratory animals, genetics, pharmacology, toxicological methods including alternatives.genetics, pharmacology, toxicological methods including alternatives.

Requirements applied on facilities Requirements applied on facilities intended for breeding, accomodation, care, intended for breeding, accomodation, care, transport and use of animals transport and use of animals - zoohygiene : space, temperature, humidity, ventilation, illumination .... - zoohygiene : space, temperature, humidity, ventilation, illumination .... - transport conditions, veterinary care and examinations, labelling and - transport conditions, veterinary care and examinations, labelling and

registry of animals, inspections, reporting registry of animals, inspections, reporting (12 Annexes related to individual species, templates for reporting ....).(12 Annexes related to individual species, templates for reporting ....).

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Animal Testing in the CR : 2009 Information Bulletine of Ministry of Agriculture

No.4 / 2010 http://eagri.cz

State (Veterinary) administration of animal experiments / Central Commission for Animal Welfare 2009Central Commission for Animal Welfare 2009

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Animal Testing in the CR : 2009 Information Bulletine of Ministry of Agriculture

No.4 / 2010 http://eagri.cz

Biological studies

Development / QC of drugs

Diagnosis of disease

Toxicology / safety evaluations

Education / training

Others

Number of animals for safety assessment of cosmetics

010

203040

5060

7080

year

nu

mb

er o

f an

imal

s

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Alternative methods in the CRintroduced originaly for In-market testing of cosmetics

In 1970sIn 1970s as a screening tool for localas a screening tool for local toxicity evaluation of consumer toxicity evaluation of consumer products / cosmetics and medical devicesproducts / cosmetics and medical devices

Reasons Reasons : scientific + ethical, economical + methodologicalNational Reference Center for Cosmetics and National Reference National Reference Center for Cosmetics and National Reference

Laboratory for Experimental Immunotoxicology at NIPHLaboratory for Experimental Immunotoxicology at NIPH Unit for Toxicology In vitro at Department of Toxicology at NIPHUnit for Toxicology In vitro at Department of Toxicology at NIPH

authorized by Ministry of Health for in-market testing of cosmetics,employ alternativesemploy alternatives on routine basis for safety and efficacy testing on routine basis for safety and efficacy testing

apply validated and valid alternatives for apply validated and valid alternatives for local toxicity testinglocal toxicity testing and safety and safety evaluation of chemicals and consumer products intended for skin/ mucosa evaluation of chemicals and consumer products intended for skin/ mucosa contact (toys, children products, MD, chemical preparations, others...)contact (toys, children products, MD, chemical preparations, others...)

testing facility for control authorities, manufacturers, NGOs , individuals... accredited according to ISO 17025, GLPaccredited according to ISO 17025, GLP principles respected principles respected

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NRL for experimental ImmunotoxicologyNRL for experimental ImmunotoxicologyUnit for Toxicology In vitro Unit for Toxicology In vitro

Department of Toxicology at NIPHDepartment of Toxicology at NIPH

Granted for projects of applied research focussed on use of alternative methods in vitro for determination of local toxicity ( phototoxicity, skin and eye irritation, skin penetration) of ingredients of consumer products (5x local projects, 2x 6.FP projects)

Participation of experts in Participation of experts in Peer Review of ESAC-ECVAMPeer Review of ESAC-ECVAM , , OECD WGs OECD WGs on on skin irritation, eye irritation validation studies, ICCVAM skin sensitizationskin irritation, eye irritation validation studies, ICCVAM skin sensitization

ESAC-ECVAMESAC-ECVAM Member Member (Head of Department )(Head of Department )Local information center for alternative methods in the CR, provide training

of other laboratories, perform comparative studiesMember of Central Commission for Animal Welfare in the CR, evaluation

of projects of experiments at Commission of Ministry of HealthCZECOPA (ECOPA) founder- registered office / executive center at NIPH

No specific budget for alternatives - included in budget of NIPHNo specific budget for alternatives - included in budget of NIPHCZECOPA : membership fees onlyCZECOPA : membership fees only

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Alternative methods performed at NIPH Regulation No.440/2008/EC - OECD TG – ISO - ESAC

Skin corrosion: B.40 EpiSkin/ EpiDerm models of epidermis (OECD 430)Skin irritation: B.46 EpiSkin, EpiDerm, SkinEthic RHE models of epidermisPhototoxicity: B.41 3T3 NRU test (OECD 432) , 3D skin modelsEye irritation : Cytotoxicity test - 3T3 NRR, Organotypic model HET-CAM, 3D cornea models : EpiOcular, HCE SkinEthic Skin absorption / penetration : B.45 (OECD 428) ex vivo porcine skinGenotoxicity / Mutagenicity :

B.13/14 Reverse mutations in bacteria (Ames test) (OECD 471) B.10 Chromozomal aberations in mammals in vitro (OECD 473) Clastogenicity + aneugenicity. In vitro micronucleus test (OECD 487)

Acute toxicity: Cytotoxicity test for starting dose for in vivo test Cytotoxicity test: EN ISO 10993-5 ISO 10993-5 for medical devices, cytotoxicity as for medical devices, cytotoxicity as

screening for local tolerance evaluation of consumer productsscreening for local tolerance evaluation of consumer products

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Directive 2010/63/EU Directive 2010/63/EU on protection of animals used for scientific purposeson protection of animals used for scientific purposes

transpositiontransposition Transposition foreseen by Ministry of Agriculture, Transposition foreseen by Ministry of Agriculture, as :as : - recognized - recognized competent authoritycompetent authority by „competence law“ for „protection of by „competence law“ for „protection of

animals“ in general and responsible for implementation.animals“ in general and responsible for implementation. - has already - has already established instrumentsestablished instruments to implement directive into the Act to implement directive into the Act

No.246/1992 Coll. on protection of animals against cruelty and Decree No.246/1992 Coll. on protection of animals against cruelty and Decree No.207/2004 Coll. on protection, breeding and use of animals for No.207/2004 Coll. on protection, breeding and use of animals for experimental purposes.experimental purposes.

Alternatively,Alternatively, Society for science on laboratory animalsSociety for science on laboratory animals suggests suggests transposition as a new law submitted by Ministry of Education : transposition as a new law submitted by Ministry of Education :

- easier transposition as law dealing only with animals used for scientific - easier transposition as law dealing only with animals used for scientific purposespurposes

- clearly separated law for laboratory animals, free from provisions on farm - clearly separated law for laboratory animals, free from provisions on farm animals, wildlife species, etc.animals, wildlife species, etc.

- majority of experiments are performed within the sector of education, i.e. - majority of experiments are performed within the sector of education, i.e. universities, scientific institutionsuniversities, scientific institutions

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Directive 2010/63/EU Directive 2010/63/EU Existing provisions / instruments – easy to update Existing provisions / instruments – easy to update

Mandatory requirement Mandatory requirement to apply to apply 3R principles3R principles, priority in replacement and , priority in replacement and use of alternative use of alternative methods in vitro.methods in vitro.

Requirements on the Requirements on the evaluation and authorization of projectsevaluation and authorization of projects involving the involving the use of animals, use of animals, qualification/competencequalification/competence of personnel. of personnel.

Requirements on the origin, breeding, marking, care and accomodation Requirements on the origin, breeding, marking, care and accomodation and killing and killing of animals, regular of animals, regular inspectionsinspections..

Requirements on Requirements on breeders, suppliers and users.breeders, suppliers and users. Animal welfare bodiesAnimal welfare bodies (committees) (committees) in facilities of breeders, suppliers and in facilities of breeders, suppliers and

users.users. ReportingReporting on experiments (number / species/ purpose). on experiments (number / species/ purpose).

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Directive 2010/63/EU Directive 2010/63/EU New provisions – new instruments neededNew provisions – new instruments needed

Classification of severity Classification of severity of procedures of procedures Reuse Reuse of animals, of animals, sharingsharing organs/tissues, rehoming/ organs/tissues, rehoming/setting freesetting free of animals of animals Retrospective assessment Retrospective assessment of projectsof projects, non-technical project summaries, non-technical project summaries „„Contribution“ Contribution“ to the development / validation of alternatives (Art.47) by MS to the development / validation of alternatives (Art.47) by MS not not

understood as financial, no extra resources identified.understood as financial, no extra resources identified. Designation of new competent authorities :Designation of new competent authorities :- Competent authorityCompetent authority (Min.of Agriculture) (Min.of Agriculture) responsible for Dir. responsible for Dir. implementationimplementation- National Committee National Committee at Min.of Agricultureat Min.of Agriculture (new) for (new) for welfare of animalswelfare of animals used for used for

scientific purposes and for scientific purposes and for evaluation/approval of projectsevaluation/approval of projects, harmonizing , harmonizing approaches of animal wefare commissions of individual sectors (Ministries) approaches of animal wefare commissions of individual sectors (Ministries)

- Contact point Contact point to provide to provide adviceadvice on regulatory on regulatory relevance/suitability of relevance/suitability of alternativesalternatives for validation, for validation, qualified laboratoryqualified laboratory to carry out validation study. to carry out validation study.

National Institute of Public HealthNational Institute of Public Health is suggested. is suggested.

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NRL / Unit for Toxicology in vitro / CZECOPA at NIPH PragueNRL / Unit for Toxicology in vitro / CZECOPA at NIPH Prague Thank you for your attentionThank you for your attention