Proefdierkunde
INHOUD
1 History ........................................................................................................................................................... 3
2 Legislation ...................................................................................................................................................... 4
3 Safety in the animal facility ........................................................................................................................... 9
Physical hazards .................................................................................................................................................. 9
Allergy ............................................................................................................................................................... 11
Chemical and protocol related hazards ............................................................................................................ 13
Zoonosis ............................................................................................................................................................ 15
4 Zoo technique of different species .............................................................................................................. 17
Mice and rats .................................................................................................................................................... 17
Hamster ............................................................................................................................................................. 19
Guinea pig ......................................................................................................................................................... 20
Rabbit ................................................................................................................................................................ 21
Xenopus............................................................................................................................................................. 22
Zebrafish............................................................................................................................................................ 23
Pigs .................................................................................................................................................................... 23
5 Housing of laboratory animals..................................................................................................................... 25
Legislation and environmental parameters ...................................................................................................... 25
Cages and environmental enrichment .............................................................................................................. 27
Special housing systems and transport ............................................................................................................. 29
Conclusion ......................................................................................................................................................... 31
6 Hygiene and microbiological standardisation ............................................................................................. 33
Hygiene ............................................................................................................................................................. 33
Medical prophylaxis ...................................................................................................................................... 33
Sanitary prophylaxis ...................................................................................................................................... 33
Pathology .......................................................................................................................................................... 35
disease........................................................................................................................................................... 35
diagnostics..................................................................................................................................................... 35
infectious cause............................................................................................................................................. 35
Disease management ........................................................................................................................................ 38
7 Anesthesia – Analgesia .................................................................................................................................. 41
General and local anesthesia ............................................................................................................................ 41
Injectable anesthesia ........................................................................................................................................ 44
Inhalation anesthesia ........................................................................................................................................ 45
Anesthesia systems and consequences ............................................................................................................ 47
Pain assessment ................................................................................................................................................ 49
Pain and Post-operative Care ............................................................................................................................ 52
8 Humane endpoints – euthanasia .................................................................................................................. 55
1
, Humane endpoints ............................................................................................................................................ 55
Euthanasia ......................................................................................................................................................... 56
9 Behavior and well-being................................................................................................................................ 59
10 Experimental techniques ........................................................................................................................ 65
11 Surgical techniques ................................................................................................................................. 69
12 Genetical standardization and transgenese ........................................................................................... 75
Genetical standardisation ................................................................................................................................. 75
Genetical uniformity ..................................................................................................................................... 76
Genetical quality control ................................................................................................................................... 82
Transgenese ...................................................................................................................................................... 83
Introduction .................................................................................................................................................. 83
Zygote injection ............................................................................................................................................. 84
Gene targeting .............................................................................................................................................. 87
14 Alternative methods ............................................................................................................................... 91
Replacement ..................................................................................................................................................... 91
Reduction .......................................................................................................................................................... 94
Refinement........................................................................................................................................................ 95
15 Feeding laboratory animals .................................................................................................................... 98
2
,1 HISTORY
Use of laboratory animals parallel with the development of medical science
- Hippocrates - Corpus Hippocraticum (± 400 BC)
o accent on anatomy, mostly descriptive character
o vivisection
- Galenus (130-201 AD)
o physiology (e.g. pigs, dogs, monkeys)
o animal was completely accepted as a model for men
- Rising of Christianity
o nearly no research done during several centuries and very little animals were used
- From the 15th century on (Renaissance)
o new urge to increase knowledge
o experiments on non-anaesthetised animals
o Cartesian view on life (Descartes, 1596-1650): conclusion animals don’t feel pain so we can use
them for experiments
- Andreas Vesalius (1514-1564)
o Belgian
o anatomy of humans
o cutting spine of animals for research
- William Harvey (1578-1657)
o animal is seen as an important source of information
o knowledge of blood circulation and valves
- Second half of the 19th century
o important increase in animal use
o discovery of anesthetics
o several important scientific discoveries
o Darwin: Origin of Species (1859) biological similarities between humans and animals
→ scientific base for the use of animals as model for humans
o Claude Bernard (1865):
▪ functioning of organs
▪ defends the use of animals in experimental work
o Emergence of microbiology
▪ Robert Koch (1884): pathogenicity of microorganism is only proven after infection
experiment with healthy laboratory animals
▪ development of vaccines and antisera (Louis Pasteur, 1880)
- After WO I
o production of pharmaca
o development of new disciplines (toxicology, immunology,…)
o strong increase in the number of laboratory animals
- Shift in type of species used
o until end 19th century: mostly dogs, cats, rabbits and pigs
o later more rodents; especially rats and mice
- Legal obligation to test compounds in animals before use in clinical studies
- 1876: first legislation concerning the use of animals in research
o Cruelty to Animals Act (England)
- Countermovements
o movement against experiments with animals, first in England, later in France (from 19th
century)
o Peter Singer (1975): Animal Liberation Movement
o Tom Regan (1983): Animal Rights Movement
→ niet te kennen voor examen
3
, 2 LEGISLATION
Numbers
Europa: 12 million animals used
Belgium (2013): 626.742 animals used
- 77% rodents (rats, mice) and 8% rabbits
- 11% fish, amphibians and reptiles
- 4% birds
- 0,6% farm animals
- 0,09 % for dogs, 0,05% cats and 0,002% primates
Use of laboratory animals is fluctuating
- Development of alternatives, ethics, legislation, more responsible use and high costs → decrease ↓
- Development of transgenic animals → increase ↑
Legislation
18 March 1986 – 18 October 1991 European Convention for the protection of vertebrate animals that are
used for experimental and other scientific purposes
14 August 1986 Belgian law concerning the protection and the well-being of animals
14 November 1993 Royal Decree concerning the protection of laboratory animals (update)
6 April 2010 New Belgian Royal Decree with even stricter rules concerning housing of
laboratory animals
September 2010 New European Directive on the protection of animals used for scientific
purposes
29 May 2013 – 10 July 2013 Royal Decree concerning transposition of the EU Directive into national
legislation
Definition of ‘Laboratory animal’
= every living vertebrate animal used or intended for laboratory experiments
- Including free living and/or reproducing larval forms (example axolotl)
- Used or intended for laboratory experiments
- Excluding other fetal or embryonic forms
What has changed?
- Include some invertebrates (example: Cephalopoda (squid, lobster), fish)
- Include mammalian embryonic forms starting from the last third of term
Definition of ‘Animal experiment’
= every use of a living vertebrate animal for experimental and other scientific purposes
- Can inflict pain, suffering, discomfort or permanent injury
- Including every treatment that has the purpose or the consequence of the birth of an animal in such a
condition
- Exception of the least painful, in modern practice accepted methods to kill or mark the animal (so-called
humane methods)
- Excluding non-experimental treatments in agriculture and in veterinary practice
What does the law say
- Pain, suffering and discomfort were not defined in legislation → now taken up in the new European
Directive
- An animal should not be used more than once in experiments that inflict serious pain and suffering
o opmerking: niet enkel pijn maar ook permanente schade moet in rekening gebracht w
vb. ‘whole body irradiation’ doet geen pijn maar wel veel schade aan beenmerg
vb. amputatie
4
INHOUD
1 History ........................................................................................................................................................... 3
2 Legislation ...................................................................................................................................................... 4
3 Safety in the animal facility ........................................................................................................................... 9
Physical hazards .................................................................................................................................................. 9
Allergy ............................................................................................................................................................... 11
Chemical and protocol related hazards ............................................................................................................ 13
Zoonosis ............................................................................................................................................................ 15
4 Zoo technique of different species .............................................................................................................. 17
Mice and rats .................................................................................................................................................... 17
Hamster ............................................................................................................................................................. 19
Guinea pig ......................................................................................................................................................... 20
Rabbit ................................................................................................................................................................ 21
Xenopus............................................................................................................................................................. 22
Zebrafish............................................................................................................................................................ 23
Pigs .................................................................................................................................................................... 23
5 Housing of laboratory animals..................................................................................................................... 25
Legislation and environmental parameters ...................................................................................................... 25
Cages and environmental enrichment .............................................................................................................. 27
Special housing systems and transport ............................................................................................................. 29
Conclusion ......................................................................................................................................................... 31
6 Hygiene and microbiological standardisation ............................................................................................. 33
Hygiene ............................................................................................................................................................. 33
Medical prophylaxis ...................................................................................................................................... 33
Sanitary prophylaxis ...................................................................................................................................... 33
Pathology .......................................................................................................................................................... 35
disease........................................................................................................................................................... 35
diagnostics..................................................................................................................................................... 35
infectious cause............................................................................................................................................. 35
Disease management ........................................................................................................................................ 38
7 Anesthesia – Analgesia .................................................................................................................................. 41
General and local anesthesia ............................................................................................................................ 41
Injectable anesthesia ........................................................................................................................................ 44
Inhalation anesthesia ........................................................................................................................................ 45
Anesthesia systems and consequences ............................................................................................................ 47
Pain assessment ................................................................................................................................................ 49
Pain and Post-operative Care ............................................................................................................................ 52
8 Humane endpoints – euthanasia .................................................................................................................. 55
1
, Humane endpoints ............................................................................................................................................ 55
Euthanasia ......................................................................................................................................................... 56
9 Behavior and well-being................................................................................................................................ 59
10 Experimental techniques ........................................................................................................................ 65
11 Surgical techniques ................................................................................................................................. 69
12 Genetical standardization and transgenese ........................................................................................... 75
Genetical standardisation ................................................................................................................................. 75
Genetical uniformity ..................................................................................................................................... 76
Genetical quality control ................................................................................................................................... 82
Transgenese ...................................................................................................................................................... 83
Introduction .................................................................................................................................................. 83
Zygote injection ............................................................................................................................................. 84
Gene targeting .............................................................................................................................................. 87
14 Alternative methods ............................................................................................................................... 91
Replacement ..................................................................................................................................................... 91
Reduction .......................................................................................................................................................... 94
Refinement........................................................................................................................................................ 95
15 Feeding laboratory animals .................................................................................................................... 98
2
,1 HISTORY
Use of laboratory animals parallel with the development of medical science
- Hippocrates - Corpus Hippocraticum (± 400 BC)
o accent on anatomy, mostly descriptive character
o vivisection
- Galenus (130-201 AD)
o physiology (e.g. pigs, dogs, monkeys)
o animal was completely accepted as a model for men
- Rising of Christianity
o nearly no research done during several centuries and very little animals were used
- From the 15th century on (Renaissance)
o new urge to increase knowledge
o experiments on non-anaesthetised animals
o Cartesian view on life (Descartes, 1596-1650): conclusion animals don’t feel pain so we can use
them for experiments
- Andreas Vesalius (1514-1564)
o Belgian
o anatomy of humans
o cutting spine of animals for research
- William Harvey (1578-1657)
o animal is seen as an important source of information
o knowledge of blood circulation and valves
- Second half of the 19th century
o important increase in animal use
o discovery of anesthetics
o several important scientific discoveries
o Darwin: Origin of Species (1859) biological similarities between humans and animals
→ scientific base for the use of animals as model for humans
o Claude Bernard (1865):
▪ functioning of organs
▪ defends the use of animals in experimental work
o Emergence of microbiology
▪ Robert Koch (1884): pathogenicity of microorganism is only proven after infection
experiment with healthy laboratory animals
▪ development of vaccines and antisera (Louis Pasteur, 1880)
- After WO I
o production of pharmaca
o development of new disciplines (toxicology, immunology,…)
o strong increase in the number of laboratory animals
- Shift in type of species used
o until end 19th century: mostly dogs, cats, rabbits and pigs
o later more rodents; especially rats and mice
- Legal obligation to test compounds in animals before use in clinical studies
- 1876: first legislation concerning the use of animals in research
o Cruelty to Animals Act (England)
- Countermovements
o movement against experiments with animals, first in England, later in France (from 19th
century)
o Peter Singer (1975): Animal Liberation Movement
o Tom Regan (1983): Animal Rights Movement
→ niet te kennen voor examen
3
, 2 LEGISLATION
Numbers
Europa: 12 million animals used
Belgium (2013): 626.742 animals used
- 77% rodents (rats, mice) and 8% rabbits
- 11% fish, amphibians and reptiles
- 4% birds
- 0,6% farm animals
- 0,09 % for dogs, 0,05% cats and 0,002% primates
Use of laboratory animals is fluctuating
- Development of alternatives, ethics, legislation, more responsible use and high costs → decrease ↓
- Development of transgenic animals → increase ↑
Legislation
18 March 1986 – 18 October 1991 European Convention for the protection of vertebrate animals that are
used for experimental and other scientific purposes
14 August 1986 Belgian law concerning the protection and the well-being of animals
14 November 1993 Royal Decree concerning the protection of laboratory animals (update)
6 April 2010 New Belgian Royal Decree with even stricter rules concerning housing of
laboratory animals
September 2010 New European Directive on the protection of animals used for scientific
purposes
29 May 2013 – 10 July 2013 Royal Decree concerning transposition of the EU Directive into national
legislation
Definition of ‘Laboratory animal’
= every living vertebrate animal used or intended for laboratory experiments
- Including free living and/or reproducing larval forms (example axolotl)
- Used or intended for laboratory experiments
- Excluding other fetal or embryonic forms
What has changed?
- Include some invertebrates (example: Cephalopoda (squid, lobster), fish)
- Include mammalian embryonic forms starting from the last third of term
Definition of ‘Animal experiment’
= every use of a living vertebrate animal for experimental and other scientific purposes
- Can inflict pain, suffering, discomfort or permanent injury
- Including every treatment that has the purpose or the consequence of the birth of an animal in such a
condition
- Exception of the least painful, in modern practice accepted methods to kill or mark the animal (so-called
humane methods)
- Excluding non-experimental treatments in agriculture and in veterinary practice
What does the law say
- Pain, suffering and discomfort were not defined in legislation → now taken up in the new European
Directive
- An animal should not be used more than once in experiments that inflict serious pain and suffering
o opmerking: niet enkel pijn maar ook permanente schade moet in rekening gebracht w
vb. ‘whole body irradiation’ doet geen pijn maar wel veel schade aan beenmerg
vb. amputatie
4
