Laboratory Animal Science – Summary
Inhoudsopgave
1 Introduction............................................................................................................................. 5
2 History..................................................................................................................................... 5
3 Legislation............................................................................................................................... 5
More changes in legislation..................................................................................................... 7
4 Safety in the animal facility..................................................................................................... 8
4.1. Physical hazards............................................................................................................... 8
Trauma................................................................................................................................. 9
Combustible material – Fire and burning..............................................................................9
Noise.................................................................................................................................... 9
Electricity............................................................................................................................. 9
Apparatus under pressure, gasses and vapors....................................................................9
Radiation.............................................................................................................................. 9
Ionising radiation............................................................................................................... 10
4.2. Allergy............................................................................................................................ 10
Screening programs........................................................................................................... 11
Design of animal facility..................................................................................................... 11
Work organisation.............................................................................................................. 11
Personal protective gear.................................................................................................... 12
4.3. Chemical and protocol related hazards..........................................................................12
Protocol-related hazards.................................................................................................... 12
4.4. Zoonosis......................................................................................................................... 13
Plague – bacteria Yersinia pestis........................................................................................ 14
Leptospirosis (Weil’s disease, canicola fever)....................................................................14
Hanta virus......................................................................................................................... 14
Lymphocytic Choriomeningitis Virus (LCMV)......................................................................14
Salmonellosis..................................................................................................................... 14
Streptobacillus moniliformis............................................................................................... 14
Cox pox disease................................................................................................................. 14
Trichophyty......................................................................................................................... 14
Herpes-B............................................................................................................................ 14
5 Zootechnique of different species......................................................................................... 15
Characteristics - Rodents................................................................................................... 15
5.1. Mice and rats.................................................................................................................. 15
Characteristics – Mice and rats.......................................................................................... 15
5.2. Hamster (Mesocricetus auratus).................................................................................... 16
Characteristics................................................................................................................... 16
5.3. Guinea pig (Cavia porcellus).......................................................................................... 17
, Characteristics................................................................................................................... 17
5.4. Rabbit (Oryctolagus cuniculus)....................................................................................... 18
Characteristics................................................................................................................... 18
5.5. African clawed frog (Xenopus laevis).............................................................................. 18
5.6. Zebrafish (Danio rerio)................................................................................................... 19
5.7. Pigs (Sus scrufa)............................................................................................................. 19
6 Housing of Laboratory animals.............................................................................................. 19
6.1. Legislation and environmental parameters....................................................................19
Environment....................................................................................................................... 20
6.2. Cages and environmental enrichment........................................................................... 22
Cage material..................................................................................................................... 22
Cage floors......................................................................................................................... 22
Water supply...................................................................................................................... 22
Feedhoppers or feedracks.................................................................................................. 23
Bedding material................................................................................................................ 23
Environmental enrichment................................................................................................. 23
6.3. Special housing systems and transport..........................................................................24
Conventional animal facility............................................................................................... 24
Semi-barrier....................................................................................................................... 24
A2-animal facility............................................................................................................... 25
Specified pathogen-free (SPF) facility................................................................................25
Transport of animals........................................................................................................... 25
7 Hygiene and microbiological standardisation........................................................................26
7.1. Hygiene.......................................................................................................................... 26
Medical prophylaxis............................................................................................................ 26
Sanitary prophylaxis.......................................................................................................... 26
Disinfection and sterilisation.............................................................................................. 27
7.2. Pathology....................................................................................................................... 28
Disease.............................................................................................................................. 28
Diagnostics........................................................................................................................ 28
Infectious causes................................................................................................................ 28
7.3. Disease Management..................................................................................................... 29
8 Anaesthesia – Analgesia........................................................................................................ 31
8.1. General anaesthesia...................................................................................................... 31
8.2. Local anaesthesia........................................................................................................... 32
8.3. Sedation......................................................................................................................... 32
8.4. Premedication................................................................................................................ 33
8.5. Muscle relaxation........................................................................................................... 33
8.6. Injectable anesthesia..................................................................................................... 33
8.7. Inhalation anesthesia..................................................................................................... 33
8.8. Anaesthesia systems and consequences.......................................................................34
, Open system...................................................................................................................... 34
Half-open system............................................................................................................... 34
Half-closed system............................................................................................................. 34
Closed system.................................................................................................................... 34
8.9. Pain assessment............................................................................................................. 34
8.10. Pain.............................................................................................................................. 35
Post-operative care............................................................................................................ 36
9 Humane Endpoints – Euthanasia........................................................................................... 36
9.1. Pharmacological-chemical methods............................................................................... 37
9.2. Mechanical-physical methods........................................................................................ 37
10 Behaviour and Well-being.................................................................................................... 38
10.1. Species-specific behavior............................................................................................. 38
10.2. Ethogram..................................................................................................................... 38
10.3. Effect of stress – Interaction with environment............................................................38
Measuring well being......................................................................................................... 40
11 Experimental Techniques..................................................................................................... 40
11.1. Identification of animals............................................................................................... 40
11.2. Administration routes................................................................................................... 41
11.3. Withdrawal routes........................................................................................................ 42
12 Surgical Techniques............................................................................................................. 42
12.1. Stages of a surgical procedure on an animal...............................................................42
12.2. Additional care during surgery of the animal...............................................................43
12.3. Surgical instruments.................................................................................................... 43
12.4. Suturing techniques..................................................................................................... 43
= placing a surgeons knot..................................................................................................... 43
13 Genetical Standardisation................................................................................................... 43
13.1. Genetical control.......................................................................................................... 43
Cryopreservation of embryo’s/sperm.................................................................................43
13.2. Background and inbred strains..................................................................................... 44
Monozygote animals.......................................................................................................... 44
Inbred strains..................................................................................................................... 44
13.3. Different inbred strains................................................................................................ 45
F1-hybrid (Filial 1).............................................................................................................. 45
Congenic strain.................................................................................................................. 45
Chromosome-substitution strain (= consomic strain)........................................................45
Recombinant inbred strain (RI-strain)................................................................................46
Recombinant congenic strain (RCS)................................................................................... 46
13.4. Outbred strains............................................................................................................ 46
14 Transgenesis (in vertebrates).............................................................................................. 46
14.1. Random insertion in heritable transgenesis.................................................................47
Zygote injection................................................................................................................. 47
, Random insertion in frogs.................................................................................................. 48
Transgene Intracytoplasmic Sperm Injection (ICSI)............................................................48
Retroviral vectors............................................................................................................... 48
Transgenic birds................................................................................................................. 49
14.2. Targeted insertion........................................................................................................ 49
14.3. Genome engineering/editing........................................................................................ 50
14.4. Cloning......................................................................................................................... 50
14.5. Chemical induced mutagenesis.................................................................................... 50
14.6. Transient knock-down of gene expression....................................................................51
15 Ethics and Laboratory Animal Use....................................................................................... 51
15.1. Philosophical background............................................................................................. 51
Scala naturae – Great chain of being (1579)......................................................................52
Zoocentrism....................................................................................................................... 52
Proportionalist viewpoint (= in between position)..............................................................53
15.2. Animal Welfare............................................................................................................. 53
Conclusion.......................................................................................................................... 54
15.3. Ethical Matrix EU directive........................................................................................... 54
16 Alternative Methods............................................................................................................ 54
16.1. Replacement................................................................................................................ 55
In vitro research................................................................................................................. 55
Use of vertebrates at early stages – embryo’s...................................................................55
Use of lower organisms...................................................................................................... 56
Audio-visual aids................................................................................................................ 56
Other alternatives.............................................................................................................. 56
16.2. Reduction..................................................................................................................... 56
Ethical consideration.......................................................................................................... 57
Experimental design (statistics)......................................................................................... 57
Pilot study.......................................................................................................................... 57
Sharing animals................................................................................................................. 57
Cryo-preservation.............................................................................................................. 57
Computer models............................................................................................................... 57
Combining.......................................................................................................................... 57
Improved information dissemination.................................................................................. 58
Telemetry........................................................................................................................... 58
Good Laboratory Practise (GLP)......................................................................................... 58
16.3. Refinement................................................................................................................... 58
17 Feeding Laboratory Animals................................................................................................ 59
17.1. Nutrient requirements.................................................................................................. 62
18 Experimental design............................................................................................................ 63
Inhoudsopgave
1 Introduction............................................................................................................................. 5
2 History..................................................................................................................................... 5
3 Legislation............................................................................................................................... 5
More changes in legislation..................................................................................................... 7
4 Safety in the animal facility..................................................................................................... 8
4.1. Physical hazards............................................................................................................... 8
Trauma................................................................................................................................. 9
Combustible material – Fire and burning..............................................................................9
Noise.................................................................................................................................... 9
Electricity............................................................................................................................. 9
Apparatus under pressure, gasses and vapors....................................................................9
Radiation.............................................................................................................................. 9
Ionising radiation............................................................................................................... 10
4.2. Allergy............................................................................................................................ 10
Screening programs........................................................................................................... 11
Design of animal facility..................................................................................................... 11
Work organisation.............................................................................................................. 11
Personal protective gear.................................................................................................... 12
4.3. Chemical and protocol related hazards..........................................................................12
Protocol-related hazards.................................................................................................... 12
4.4. Zoonosis......................................................................................................................... 13
Plague – bacteria Yersinia pestis........................................................................................ 14
Leptospirosis (Weil’s disease, canicola fever)....................................................................14
Hanta virus......................................................................................................................... 14
Lymphocytic Choriomeningitis Virus (LCMV)......................................................................14
Salmonellosis..................................................................................................................... 14
Streptobacillus moniliformis............................................................................................... 14
Cox pox disease................................................................................................................. 14
Trichophyty......................................................................................................................... 14
Herpes-B............................................................................................................................ 14
5 Zootechnique of different species......................................................................................... 15
Characteristics - Rodents................................................................................................... 15
5.1. Mice and rats.................................................................................................................. 15
Characteristics – Mice and rats.......................................................................................... 15
5.2. Hamster (Mesocricetus auratus).................................................................................... 16
Characteristics................................................................................................................... 16
5.3. Guinea pig (Cavia porcellus).......................................................................................... 17
, Characteristics................................................................................................................... 17
5.4. Rabbit (Oryctolagus cuniculus)....................................................................................... 18
Characteristics................................................................................................................... 18
5.5. African clawed frog (Xenopus laevis).............................................................................. 18
5.6. Zebrafish (Danio rerio)................................................................................................... 19
5.7. Pigs (Sus scrufa)............................................................................................................. 19
6 Housing of Laboratory animals.............................................................................................. 19
6.1. Legislation and environmental parameters....................................................................19
Environment....................................................................................................................... 20
6.2. Cages and environmental enrichment........................................................................... 22
Cage material..................................................................................................................... 22
Cage floors......................................................................................................................... 22
Water supply...................................................................................................................... 22
Feedhoppers or feedracks.................................................................................................. 23
Bedding material................................................................................................................ 23
Environmental enrichment................................................................................................. 23
6.3. Special housing systems and transport..........................................................................24
Conventional animal facility............................................................................................... 24
Semi-barrier....................................................................................................................... 24
A2-animal facility............................................................................................................... 25
Specified pathogen-free (SPF) facility................................................................................25
Transport of animals........................................................................................................... 25
7 Hygiene and microbiological standardisation........................................................................26
7.1. Hygiene.......................................................................................................................... 26
Medical prophylaxis............................................................................................................ 26
Sanitary prophylaxis.......................................................................................................... 26
Disinfection and sterilisation.............................................................................................. 27
7.2. Pathology....................................................................................................................... 28
Disease.............................................................................................................................. 28
Diagnostics........................................................................................................................ 28
Infectious causes................................................................................................................ 28
7.3. Disease Management..................................................................................................... 29
8 Anaesthesia – Analgesia........................................................................................................ 31
8.1. General anaesthesia...................................................................................................... 31
8.2. Local anaesthesia........................................................................................................... 32
8.3. Sedation......................................................................................................................... 32
8.4. Premedication................................................................................................................ 33
8.5. Muscle relaxation........................................................................................................... 33
8.6. Injectable anesthesia..................................................................................................... 33
8.7. Inhalation anesthesia..................................................................................................... 33
8.8. Anaesthesia systems and consequences.......................................................................34
, Open system...................................................................................................................... 34
Half-open system............................................................................................................... 34
Half-closed system............................................................................................................. 34
Closed system.................................................................................................................... 34
8.9. Pain assessment............................................................................................................. 34
8.10. Pain.............................................................................................................................. 35
Post-operative care............................................................................................................ 36
9 Humane Endpoints – Euthanasia........................................................................................... 36
9.1. Pharmacological-chemical methods............................................................................... 37
9.2. Mechanical-physical methods........................................................................................ 37
10 Behaviour and Well-being.................................................................................................... 38
10.1. Species-specific behavior............................................................................................. 38
10.2. Ethogram..................................................................................................................... 38
10.3. Effect of stress – Interaction with environment............................................................38
Measuring well being......................................................................................................... 40
11 Experimental Techniques..................................................................................................... 40
11.1. Identification of animals............................................................................................... 40
11.2. Administration routes................................................................................................... 41
11.3. Withdrawal routes........................................................................................................ 42
12 Surgical Techniques............................................................................................................. 42
12.1. Stages of a surgical procedure on an animal...............................................................42
12.2. Additional care during surgery of the animal...............................................................43
12.3. Surgical instruments.................................................................................................... 43
12.4. Suturing techniques..................................................................................................... 43
= placing a surgeons knot..................................................................................................... 43
13 Genetical Standardisation................................................................................................... 43
13.1. Genetical control.......................................................................................................... 43
Cryopreservation of embryo’s/sperm.................................................................................43
13.2. Background and inbred strains..................................................................................... 44
Monozygote animals.......................................................................................................... 44
Inbred strains..................................................................................................................... 44
13.3. Different inbred strains................................................................................................ 45
F1-hybrid (Filial 1).............................................................................................................. 45
Congenic strain.................................................................................................................. 45
Chromosome-substitution strain (= consomic strain)........................................................45
Recombinant inbred strain (RI-strain)................................................................................46
Recombinant congenic strain (RCS)................................................................................... 46
13.4. Outbred strains............................................................................................................ 46
14 Transgenesis (in vertebrates).............................................................................................. 46
14.1. Random insertion in heritable transgenesis.................................................................47
Zygote injection................................................................................................................. 47
, Random insertion in frogs.................................................................................................. 48
Transgene Intracytoplasmic Sperm Injection (ICSI)............................................................48
Retroviral vectors............................................................................................................... 48
Transgenic birds................................................................................................................. 49
14.2. Targeted insertion........................................................................................................ 49
14.3. Genome engineering/editing........................................................................................ 50
14.4. Cloning......................................................................................................................... 50
14.5. Chemical induced mutagenesis.................................................................................... 50
14.6. Transient knock-down of gene expression....................................................................51
15 Ethics and Laboratory Animal Use....................................................................................... 51
15.1. Philosophical background............................................................................................. 51
Scala naturae – Great chain of being (1579)......................................................................52
Zoocentrism....................................................................................................................... 52
Proportionalist viewpoint (= in between position)..............................................................53
15.2. Animal Welfare............................................................................................................. 53
Conclusion.......................................................................................................................... 54
15.3. Ethical Matrix EU directive........................................................................................... 54
16 Alternative Methods............................................................................................................ 54
16.1. Replacement................................................................................................................ 55
In vitro research................................................................................................................. 55
Use of vertebrates at early stages – embryo’s...................................................................55
Use of lower organisms...................................................................................................... 56
Audio-visual aids................................................................................................................ 56
Other alternatives.............................................................................................................. 56
16.2. Reduction..................................................................................................................... 56
Ethical consideration.......................................................................................................... 57
Experimental design (statistics)......................................................................................... 57
Pilot study.......................................................................................................................... 57
Sharing animals................................................................................................................. 57
Cryo-preservation.............................................................................................................. 57
Computer models............................................................................................................... 57
Combining.......................................................................................................................... 57
Improved information dissemination.................................................................................. 58
Telemetry........................................................................................................................... 58
Good Laboratory Practise (GLP)......................................................................................... 58
16.3. Refinement................................................................................................................... 58
17 Feeding Laboratory Animals................................................................................................ 59
17.1. Nutrient requirements.................................................................................................. 62
18 Experimental design............................................................................................................ 63
