Alternative methods
contemporary and future situation
in the Czech Republic
Dagmar Jírová,M.D., Ph.D.
Department of Toxicology
National Institute of Public Health
Praha, CR
Protection of Animals Used for Experimental Purposes
legislation in the CR
Directive 86/609/EEC, on protection of animals used for
experimental purposes
is implemented in
Act No.246/1992 Sb. , on the protection of animals against cruelty
Decree No. 207 / 2004 / Coll., on protection, breeding and use of
animals for experimental purposes
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Act No.246/1992 Sb. ,
on the protection of animals against cruelty
Regulating :
 definition of cruelty, possible purposes / procedures to kill animals
 protection of animals during transport
 protection of farm animals
 protection of domestic animals / breeding of special interest
 protection of wildlife species of animals
 protection of animals used for experimental purposes : article 15 - 18
 competent authorities responsible for protection of animals :
Ministry of Agriculture, Central Commission for Animal Welfare (consultant
body to the Minister), State Veterinary Administration, Municipal Authorities
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Act No.246/1992 Sb. ,
article 15 - 18
animals used for experimental purposes
Possible purposes to conduct experiments on animals, including :
- ban on testing of cosmetics and their ingredients, ..... weapons
- ban on use of abandoned animals , .... animals of endangered species
- priority to use animals bred for experimental purposes
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Experiment can only be performed, if proved that no alternative methods are
available ; unavoidable pain and distress should be minimized.
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Obligation to prepare project of the experiment , it has to be approved by
Animal Wefare Commission of the respective Ministry
Requirements on qualification of personnel conducting experiments
Rules on breeding, care, accomodation and transport of experimental
animals, on accreditation of breeders, suppliers and users, on animal
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welfare committees of these facilities, reports on number and species used.
Decree No. 207 / 2004 Coll.,
on protection, breeding and use of animals for
experimental purposes
25 mandatory items to be addressed in project of the experiment, including
reasoning for unavoidability of the experiment, absence of alternatives,
justification of species selection, number of animals, possible anesthesia,
qualification of personnel, laboratory eqiupment .....
Requirements on application for accreditation of the facility.
Requirements on qualification of personnel conducting experiments : obligatory
examination in elementary anatomy and physiology of laboratory animals,
genetics, pharmacology, toxicological methods including alternatives.
Requirements applied on facilities intended for breeding, accomodation, care,
transport and use of animals
- zoohygiene : space, temperature, humidity, ventilation, illumination ....
- transport conditions, veterinary care and examinations, labelling and
registry of animals, inspections, reporting
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(12 Annexes related to individual species, templates for reporting ....).
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 2009
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Animal Testing in the CR : 2009
Information Bulletine of Ministry of Agriculture
No.4 / 2010
http://eagri.cz
80
70
60
50
40
30
20
10
0
20
09
20
08
20
07
20
06
20
05
20
04
20
03
20
02
20
01
Development / QC
of drugs
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00
19
99
number of animals
Number of animals for safety assessment of
cosmetics
year
Biological
studies
Diagnosis
of disease
Toxicology /
safety
evaluations
Education /
training
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Others
Alternative methods in the CR
introduced originaly for In-market testing of cosmetics
In 1970s as a screening tool for local toxicity evaluation of consumer
products / cosmetics and medical devices
Reasons : scientific + ethical, economical + methodological
National Reference Center for Cosmetics and National Reference
Laboratory for Experimental Immunotoxicology at NIPH
Unit for Toxicology In vitro at Department of Toxicology at NIPH
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authorized by Ministry of Health for in-market testing of cosmetics,
employ alternatives on routine basis for safety and efficacy testing
apply validated and valid alternatives for local toxicity testing and safety
evaluation of chemicals and consumer products intended for skin/ mucosa
contact (toys, children products, MD, chemical preparations, others...)
testing facility for control authorities, manufacturers, NGOs , individuals...
accredited according to ISO 17025, GLP principles respected
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NRL for experimental Immunotoxicology
Unit for Toxicology In vitro
Department 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 Peer Review of ESAC-ECVAM , OECD WGs on
skin irritation, eye irritation validation studies, ICCVAM skin sensitization
ESAC-ECVAM Member (Head of Department )
Local information center for alternative methods in the CR, provide training
of other laboratories, perform comparative studies
Member of Central Commission for Animal Welfare in the CR, evaluation
of projects of experiments at Commission of Ministry of Health
CZECOPA (ECOPA) founder- registered office / executive center at NIPH
No specific budget for alternatives - included in budget of NIPH
CZECOPA : 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 epidermis
Phototoxicity: B.41 3T3 NRU test (OECD 432) , 3D skin models
Eye 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 skin
Genotoxicity / 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 for medical devices, cytotoxicity as 10
screening for local tolerance evaluation of consumer products
Directive 2010/63/EU
on protection of animals used for scientific purposes
transposition
 Transposition foreseen by Ministry of Agriculture, as :
- recognized competent authority by „competence law“ for „protection of
animals“ in general and responsible for implementation.
- has already established instruments to implement directive into the Act
No.246/1992 Coll. on protection of animals against cruelty and Decree
No.207/2004 Coll. on protection, breeding and use of animals for
experimental purposes.
 Alternatively, Society for science on laboratory animals suggests
transposition as a new law submitted by Ministry of Education :
- easier transposition as law dealing only with animals used for scientific
purposes
- clearly separated law for laboratory animals, free from provisions on farm
animals, wildlife species, etc.
- majority of experiments are performed within the sector of education, i.e.
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universities, scientific institutions
Directive 2010/63/EU
Existing provisions / instruments – easy to update
 Mandatory requirement to apply 3R principles, priority in replacement and
use of alternative methods in vitro.
 Requirements on the evaluation and authorization of projects involving the
use of animals, qualification/competence of personnel.
 Requirements on the origin, breeding, marking, care and accomodation
and killing of animals, regular inspections.
 Requirements on breeders, suppliers and users.
 Animal welfare bodies (committees) in facilities of breeders, suppliers and
users.
 Reporting on experiments (number / species/ purpose).
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Directive 2010/63/EU
New provisions – new instruments needed
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Classification of severity of procedures
Reuse of animals, sharing organs/tissues, rehoming/setting free of animals
Retrospective assessment of projects, non-technical project summaries
„Contribution“ to the development / validation of alternatives (Art.47) by MS
not understood as financial, no extra resources identified.
Designation of new competent authorities :
Competent authority (Min.of Agriculture) responsible for Dir. implementation
National Committee at Min.of Agriculture (new) for welfare of animals used
for scientific purposes and for evaluation/approval of projects, harmonizing
approaches of animal wefare commissions of individual sectors (Ministries)
Contact point to provide advice on regulatory relevance/suitability of
alternatives for validation, qualified laboratory to carry out validation study.
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National Institute of Public Health is suggested.
NRL / Unit for Toxicology in vitro / CZECOPA at NIPH Prague
Thank you for your attention
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