Course on Laboratory animal science (NWI-BM024D)
Jessie Hendricks, Radboud University Nijmegen
2020
1
,5-10-2020 8:30-9:15 Introduction to the course (Merel Ritskes-Hoitinga)
Main aims of the course:
Make an ethical argumentation and decision on whether an animal experiment is required or not
Design an animal experiment that ends in scientifically reliable and useful results, considering animal
welfare.
5-10-2020 9:30-10:30 Introduction in laboratory animal science (Merel Ritskes-Hoitinga)
We have a reproducibility crisis in research (50-90%) and a translational crisis in animal research.
Animal models are limited in their ability to translate to humans: of the drugs that have proved
promising in animal trials, 86-90% fail in human trials.
What is laboratory animal science? A multidisciplinary science contributing to:
- Quality science and animal studies
- Health and wellbeing laboratory animals
- Public acceptance animal studies
Softenon was tested in pregnant rats but the pups died in the uterus were not detected
About 10% of the mouse weight is blood, and you can take about 10% of the total blood. So you can
collect 1% of the body weight.
Rodents need sterile surgery.
Why animal studies? To enhance health and well-being of humans.
3Rs are central
Reduction, refinement, replacement
Vermindering, verfijning, vervanging
Systematic Reviews (SR) lead to better 3R implementation because existing relevant 3R information
is not found/used. Preclinical SRs benefits:
• More evidence-based choice of models
• Transparancy on quality of reporting
• Translational transparancy: Cohen BMJ 2018
• Identifying relevant research questions
• Implementing 3Rs, e.g. prevent unnecessary duplication
• Improving project planning of animal ánd human studies
SR includes the quality of publications: Quality score and risk of bias
Quality of reporting: ARRIVE and PREPARE
European Quality in Preclinical Data (EQIPD) = Quality system development for preclinical (animal)
studies for labs in industry and academia
5-10-2020 10:45-11:30 Explanation of group assignment
AWB = animal welfare body
CCD = central committee animal experiments
DEC = animal ethical committee
2
,5-10-2020 Legislation (online lecture, Ivo Tiebosch)
This module provides a relevant level of understanding of the national and international legal and
regulatory framework within which projects involving animals are constructed and managed and of
the legal responsibilities of the people involved, i.e. those carrying out procedures on animals;
designing procedures and projects; taking care of animals; or killing animals, and may cover other
relevant legislation.
Part 1: Law and regulations (13 min)
Intent of legislation: Protection of the animal, because the animal has intrinsic value (Art 1a).
According to article 1d: No, unless…. (3R principle).
A brief history
1992: Health and welfare law for animals (Netherlands)
2013: Law animals (Netherlands)
Current relevant legislation:
2014: revised experiments on animals act (Netherlands)
Experiments on Animals Decree
Experiments on Animal Regulation
Directive 2010/63/EU (Europe)
Annex I-VI within directive
Guidance documents
Other relevant laws
- Law of Nature Preservation; protection of species: Licence for field work, relevant for animal
experiments in the wild
- Law on Animals: Transport, Weaning, Mutilation (dehorning, toe clipping), Housing (Decree
on keepers of animals)
- Law on Environmental Pollution: GMO
Part 2: what is an animal experiment
- Species, life stages: Vertebrate species, squids. For human embryos: 3th stage of pregnancy.
For fish: when they hatch and feed themselves.
- Goal of experiment
- Pain, suffering: injecting is the lower threshold
Definition of an animal experiment: Any use, invasive or non-invasive, of an animal for experimental
or other scientific purposes, with known or unknown outcome, or educational purposes, which may
cause the animal a level of pain, suffering, distress or lasting harm equivalent to, or higher than, that
caused by the introduction of a needle in accordance with good veterinary practice.
Three R’s should be met.
According to the European directive, killing of animals solely for the use of their organs or tissues is
excluded. In the Netherlands, this is also an animal experiment.
Part 3: Involved parties, procedures and control
Nationally & locally
CA (CCD; centrale commissie dierproeven) is the party that forms final ethical judgement and gives
project licence.
NVWA: institutional licence, external supervision, yearly registration
NCad: Advice to minister, CCD, AWBs, disperse best practices
Responsible researcher:
- Person responsible for set-up of project and animal experiment
- MSc with 500 hours of education in basic biology and LAS course
- Species-specific knowledge
Application for project licence:
3
, - Non-technical summary: simple Dutch, publicly available
- Project proposal: background, goal, importance, research strategy
- Appendices animal experiments: type of procedures, number of animals, discomfort, HEP,
detailed enough for ethical judgement.
Animal welfare body: Animal welfare, science quality, legal demands. Give advice and provide
supervision.
Animal Ethics Committee: independent committee, ethical and scientific evaluation, Advice to
Competent Authority (CCD)
Competent Authority: centrale commissie dierproeven:
Ethical judgement and Grant project licence
Part 4: Local execution of an animal experiment
Housing, discomfort, humane endpoints, euthanasia
Working protocol: Fits exactly within project licence and includes details of experiments, method of
housing, level of discomfort, humane end points, method of killing, agreements/responsibilities,
amendments.
Housing: Annex III of Dir. 2010/63. Differs from other animal (welfare) legislation! Social housing,
cage size, bedding, enrichment, climate
Level of discomfort: Non-recovery, mild, moderate (e.g. experimental diseases), severe
Avoid suffering: use anaesthesia, use analgesics, unless strictly motivated. Spontaneous death should
be avoided and replaced by humane endpoints.
Humane endpoints: reasons to stop the animal experiment
1. The result have been achieved
2. The animal suffers severely and unintentional
3. The animal is no longer suitable as a model and/or results can no longer be achieved
A common misunderstanding is that animals should be killed at a humane endpoint.
General parameters of severe and unintentional suffering: deviant behaviour, body weight,
locomotion/mobility, food and water intake, general body condition. Or specific clinical signs: tumour
size, joint swelling, wound infection, paralysis, etc.
Clear indicators are important for the animals discomfort, but also for the researchers:
Animal in bad condition leads to unreliable results. Spontaneous death loses the possibility to acquire
data from the animal.
Method of killing: confirmation of cessation of circulation, destruction of brain, dislocation of neck,
exsanguination, confirmation of rigor mortis.
Local personnel
Persons taking care of animals and carrying out procedures:
- Animal caretaker (MBO level 3)
- Animal technician (HBMO level 4, HBO)
- Veterinarian
- Researcher (Art. 9)
Welfare log contains description of clinical signs. Mostly done by caretaker, monitoring daily.
Audits by Animal Welfare Body: execution according to project licence and working protocol, welfare
log, competence, communication, housing/care/climate.
External supervision by NVWA: regular inspection visits (usually unannounced). They can charge
disciplinary actions.
Final registration
After the animal experiment: welfare evaluation (course of animal experiment, actual level of
discomfort, any particulars, learning outcome), retrospective evaluation, yearly registration.
4
Jessie Hendricks, Radboud University Nijmegen
2020
1
,5-10-2020 8:30-9:15 Introduction to the course (Merel Ritskes-Hoitinga)
Main aims of the course:
Make an ethical argumentation and decision on whether an animal experiment is required or not
Design an animal experiment that ends in scientifically reliable and useful results, considering animal
welfare.
5-10-2020 9:30-10:30 Introduction in laboratory animal science (Merel Ritskes-Hoitinga)
We have a reproducibility crisis in research (50-90%) and a translational crisis in animal research.
Animal models are limited in their ability to translate to humans: of the drugs that have proved
promising in animal trials, 86-90% fail in human trials.
What is laboratory animal science? A multidisciplinary science contributing to:
- Quality science and animal studies
- Health and wellbeing laboratory animals
- Public acceptance animal studies
Softenon was tested in pregnant rats but the pups died in the uterus were not detected
About 10% of the mouse weight is blood, and you can take about 10% of the total blood. So you can
collect 1% of the body weight.
Rodents need sterile surgery.
Why animal studies? To enhance health and well-being of humans.
3Rs are central
Reduction, refinement, replacement
Vermindering, verfijning, vervanging
Systematic Reviews (SR) lead to better 3R implementation because existing relevant 3R information
is not found/used. Preclinical SRs benefits:
• More evidence-based choice of models
• Transparancy on quality of reporting
• Translational transparancy: Cohen BMJ 2018
• Identifying relevant research questions
• Implementing 3Rs, e.g. prevent unnecessary duplication
• Improving project planning of animal ánd human studies
SR includes the quality of publications: Quality score and risk of bias
Quality of reporting: ARRIVE and PREPARE
European Quality in Preclinical Data (EQIPD) = Quality system development for preclinical (animal)
studies for labs in industry and academia
5-10-2020 10:45-11:30 Explanation of group assignment
AWB = animal welfare body
CCD = central committee animal experiments
DEC = animal ethical committee
2
,5-10-2020 Legislation (online lecture, Ivo Tiebosch)
This module provides a relevant level of understanding of the national and international legal and
regulatory framework within which projects involving animals are constructed and managed and of
the legal responsibilities of the people involved, i.e. those carrying out procedures on animals;
designing procedures and projects; taking care of animals; or killing animals, and may cover other
relevant legislation.
Part 1: Law and regulations (13 min)
Intent of legislation: Protection of the animal, because the animal has intrinsic value (Art 1a).
According to article 1d: No, unless…. (3R principle).
A brief history
1992: Health and welfare law for animals (Netherlands)
2013: Law animals (Netherlands)
Current relevant legislation:
2014: revised experiments on animals act (Netherlands)
Experiments on Animals Decree
Experiments on Animal Regulation
Directive 2010/63/EU (Europe)
Annex I-VI within directive
Guidance documents
Other relevant laws
- Law of Nature Preservation; protection of species: Licence for field work, relevant for animal
experiments in the wild
- Law on Animals: Transport, Weaning, Mutilation (dehorning, toe clipping), Housing (Decree
on keepers of animals)
- Law on Environmental Pollution: GMO
Part 2: what is an animal experiment
- Species, life stages: Vertebrate species, squids. For human embryos: 3th stage of pregnancy.
For fish: when they hatch and feed themselves.
- Goal of experiment
- Pain, suffering: injecting is the lower threshold
Definition of an animal experiment: Any use, invasive or non-invasive, of an animal for experimental
or other scientific purposes, with known or unknown outcome, or educational purposes, which may
cause the animal a level of pain, suffering, distress or lasting harm equivalent to, or higher than, that
caused by the introduction of a needle in accordance with good veterinary practice.
Three R’s should be met.
According to the European directive, killing of animals solely for the use of their organs or tissues is
excluded. In the Netherlands, this is also an animal experiment.
Part 3: Involved parties, procedures and control
Nationally & locally
CA (CCD; centrale commissie dierproeven) is the party that forms final ethical judgement and gives
project licence.
NVWA: institutional licence, external supervision, yearly registration
NCad: Advice to minister, CCD, AWBs, disperse best practices
Responsible researcher:
- Person responsible for set-up of project and animal experiment
- MSc with 500 hours of education in basic biology and LAS course
- Species-specific knowledge
Application for project licence:
3
, - Non-technical summary: simple Dutch, publicly available
- Project proposal: background, goal, importance, research strategy
- Appendices animal experiments: type of procedures, number of animals, discomfort, HEP,
detailed enough for ethical judgement.
Animal welfare body: Animal welfare, science quality, legal demands. Give advice and provide
supervision.
Animal Ethics Committee: independent committee, ethical and scientific evaluation, Advice to
Competent Authority (CCD)
Competent Authority: centrale commissie dierproeven:
Ethical judgement and Grant project licence
Part 4: Local execution of an animal experiment
Housing, discomfort, humane endpoints, euthanasia
Working protocol: Fits exactly within project licence and includes details of experiments, method of
housing, level of discomfort, humane end points, method of killing, agreements/responsibilities,
amendments.
Housing: Annex III of Dir. 2010/63. Differs from other animal (welfare) legislation! Social housing,
cage size, bedding, enrichment, climate
Level of discomfort: Non-recovery, mild, moderate (e.g. experimental diseases), severe
Avoid suffering: use anaesthesia, use analgesics, unless strictly motivated. Spontaneous death should
be avoided and replaced by humane endpoints.
Humane endpoints: reasons to stop the animal experiment
1. The result have been achieved
2. The animal suffers severely and unintentional
3. The animal is no longer suitable as a model and/or results can no longer be achieved
A common misunderstanding is that animals should be killed at a humane endpoint.
General parameters of severe and unintentional suffering: deviant behaviour, body weight,
locomotion/mobility, food and water intake, general body condition. Or specific clinical signs: tumour
size, joint swelling, wound infection, paralysis, etc.
Clear indicators are important for the animals discomfort, but also for the researchers:
Animal in bad condition leads to unreliable results. Spontaneous death loses the possibility to acquire
data from the animal.
Method of killing: confirmation of cessation of circulation, destruction of brain, dislocation of neck,
exsanguination, confirmation of rigor mortis.
Local personnel
Persons taking care of animals and carrying out procedures:
- Animal caretaker (MBO level 3)
- Animal technician (HBMO level 4, HBO)
- Veterinarian
- Researcher (Art. 9)
Welfare log contains description of clinical signs. Mostly done by caretaker, monitoring daily.
Audits by Animal Welfare Body: execution according to project licence and working protocol, welfare
log, competence, communication, housing/care/climate.
External supervision by NVWA: regular inspection visits (usually unannounced). They can charge
disciplinary actions.
Final registration
After the animal experiment: welfare evaluation (course of animal experiment, actual level of
discomfort, any particulars, learning outcome), retrospective evaluation, yearly registration.
4
