LABORATORY
ANIMAL SCIENCE
(LAS) - RODENTS
UAntwerpen
,TABLE OF CONTENTS
CH 0: INTRODUCTION ..............................................................................................................5
CH1: LEGISLATION (C. VAN GINNIKEN) ..................................................................................6
1. LEGISLATION ON WORKING WITH LABORATORY ANIMALS ...................................................................... 6
2. LABORATORY ANIMAL – DEFINITION ............................................................................................... 8
3. LABORATORY ANIMAL – ORIGIN ..................................................................................................... 8
4. LABORATORY ANIMAL – IDENTIFICATION ........................................................................................ 10
5. ANIMAL EXPERIMENT – PURPOSES ............................................................................................... 11
6. ETHICAL COMMITTEE ............................................................................................................... 12
7. ANIMAL WELFARE UNIT/BODY ..................................................................................................... 13
8. USER, SUPPLIER, BREEDER ........................................................................................................ 13
9. ANIMAL HUSBANDRY ............................................................................................................... 14
10. PERSONNEL AND THEIR RESPONSABILITIES (!) ................................................................................. 15
11. AT THE END OF THE EXPERIMENT – OUTCOMES ................................................................................ 17
CH2: PRINCIPLES OF ANIMAL HANDLING AND RESTRAINT (D. VAN DAM) .............................. 19
1. THE 3HS-MODULES ................................................................................................................ 19
2. ANIMAL HANDLING ................................................................................................................. 19
3. NON-AVERSIVE / REFINED HANDLING OF MICE ................................................................................ 21
4. RAT HANDLING / TOUCHING ...................................................................................................... 25
5. PICKING UP ANIMALS ............................................................................................................... 27
6. IMMOBILISATION OF ANIMALS..................................................................................................... 28
CH3: BIOLOGY OF RODENTS (C. VAN GINNIKEN) .................................................................. 33
1. MOUSE – NUMBERS (BELGIUM / FLANDERS) ................................................................................... 33
2. MOUSE – BIOLOGY ................................................................................................................. 33
3. ANIMAL HUSBANDRY CONSIDERATIONS (GENERAL) ........................................................................ 36
4. MOUSE HUSBANDRY............................................................................................................... 37
5. MOUSE – ENVIRONMENTAL PARAMETERS ..................................................................................... 41
6. MOUSE – ANATOMICAL & PHYSIOLOGICAL FEATURES ...................................................................... 42
7. MOUSE – NUTRITION & REPRODUCTION ...................................................................................... 46
8. MOUSE – REPRODUCTIVE AND URINARY SYSTEM ............................................................................. 47
9. MOUSE – FERTILITY CYCLE, BREEDING & REPRODUCTION ................................................................ 48
10. MOUSE – DEVELOPMENT & BREEDING PROTOCOLS ........................................................................ 51
11. RAT – NUMBERS .................................................................................................................... 54
12. RAT – BIOLOGY...................................................................................................................... 54
13. RAT – HUSBANDRY ................................................................................................................. 55
14. RAT – ENVIRONMENTAL PARAMETERS, ANATOMY & REPRODUCTION .................................................... 56
1
,15. RAT – BREEDING & REPRODUCTION ............................................................................................ 57
CH4: ANIMAL WELFARE (D. VAN DAM) .................................................................................. 59
1. IMPACT ON ANIMAL WELFARE IN RESEARCH .................................................................................. 59
2. ANIMAL WELFARE AND SENTIENCE ............................................................................................. 60
3. CONTROLLABILITY AND PREDICTABILITY ........................................................................................ 64
4. FREQUENCY OF STRESSORS & HABITUATION .................................................................................. 66
5. REGULATORY RANGE & ADAPTIVE CAPACITY ................................................................................. 67
6. STRESS, DISTRESS & ANIMAL WELFARE ....................................................................................... 68
7. HABITUATION VS. SENSITIZATION................................................................................................ 69
8. STRESS THROUGHOUT THE LIFE CYCLE (EXAMPLES) ........................................................................ 69
CH5: BEHAVIORAL EVALUATION (VAN DAM) ......................................................................... 75
1. GOAL OF BEHAVIORAL EVALUATION ............................................................................................ 75
2. MULTI-TIERED EVALUATION PROCESS ......................................................................................... 75
3. TIER 1: GENERAL HEALTH AND NEUROLOGICAL REFLEXES (DETAILS) ................................................... 76
4. SENSORY AND MOTOR ABILITIES ................................................................................................ 81
5. SPECIFIC BEHAVIORAL DOMAINS (3RD TIER): SOCIAL BEHAVIORS ....................................................... 87
6. FEAR AND ANXIETY ................................................................................................................. 91
7. CIRCADIAN RHYTHM AND SLEEP ................................................................................................ 92
8. LEARNING AND MEMORY (COGNITIVE FUNCTION TESTS) .................................................................. 93
9. DEPRESSION-RELATED SYMPTOMS ............................................................................................. 97
CH6: MINIMALLY INVASIVE PROCEDURES (C. VAN GINNIKEN) .............................................. 98
1. IDENTIFICATION / MARKING OF ANIMALS ....................................................................................... 98
2. ADMINISTRATION OF SUBSTANCES............................................................................................. 103
3. COLLECTION OF BODY FLUIDS .................................................................................................. 108
CH7: ANESTHESIA AND ANALGESIA (C. VAN GINNEKEN) .................................................... 117
1. PRE-OPERATIVE MANAGEMENT ................................................................................................ 117
2. PRE-OPERATIVE: FASTING ...................................................................................................... 120
3. PERI-OPERATIVE PERIOD – MANAGEMENT................................................................................... 122
4. DEFINITIONS ....................................................................................................................... 123
5. ANESTHESIA ....................................................................................................................... 124
6. THE TRIAD OF ANESTHESIA ..................................................................................................... 124
7. ANESTHESIA – PHASES .......................................................................................................... 125
8. ANESTHETIC DEPTH .............................................................................................................. 125
9. INHALATION ANESTHETICS...................................................................................................... 127
10. INJECTABLE ANAESTHETICS ..................................................................................................... 132
11. ANAESTHESIA – PHARMACA .................................................................................................... 137
2
,12. OBSERVATIONAL MONITORING DURING ANAESTHESIA ..................................................................... 138
13. TROUBLESHOOTING .............................................................................................................. 139
14. POST-OPERATIVE MANAGEMENT ............................................................................................... 140
15. PAIN & ANALGESIA ............................................................................................................... 141
16. PRE-EMPTIVE ANALGESIA ....................................................................................................... 141
17. PAIN MANAGEMENT – ANALGESICS ........................................................................................... 142
18. PAIN ASSESSMENT ............................................................................................................... 144
CH8: PAIN AND WELFARE ASSESSMENT (D. VAN DAM) ....................................................... 146
1. PAIN AND WELFARE ASSESSMENT ............................................................................................ 146
2. WELFARE ASSESSMENT ......................................................................................................... 146
3. WELFARE INDICATORS........................................................................................................... 146
4. TUTORIAL: ANIMAL HEALTH AND WELFARE (AHWLA) ................................................................... 147
5. BODY CONDITION ................................................................................................................ 147
6. BODY SHAPE AND MALFORMATIONS ......................................................................................... 148
7. POSTURE ........................................................................................................................... 149
8. FUR QUALITY ...................................................................................................................... 150
9. EYES AND MOUTH ................................................................................................................ 150
10. SKIN AND MUCOUS MEMBRANES ............................................................................................. 151
11. TAIL .................................................................................................................................. 152
12. BEHAVIOURAL ASPECTS ......................................................................................................... 152
13. PAIN ASSESSMENT ............................................................................................................... 154
14. GRIMACE SCALE (FACIAL EXPRESSION ANALYSIS)......................................................................... 159
15. ESSENTIAL KNOWLEDGE FOR WELFARE ASSESSMENT .................................................................... 161
16. WELFARE ASSESSMENT IN GENETICALLY MODIFIED (GM) RODENTS .................................................. 162
CH9: HUMANE KILLING ........................................................................................................ 164
1. LEGISLATION ON HUMANE KILLING ........................................................................................... 164
2. REQUIREMENTS FOR HUMANE KILLING ...................................................................................... 164
3. METHODS FOR HUMANE KILLING (EXAM) ................................................................................. 164
4. CONFIRMATION OF DEATH...................................................................................................... 167
5. HUMANE ENDPOINTS ............................................................................................................ 167
CH10: DISEASES (C. VAN GINNEKEN) ............................................................................... 168
1. HOW MICROORGANISMS ENTER THE ANIMAL UNIT ......................................................................... 168
2. HEALTH MONITORING (HM) ................................................................................................... 169
3. MAJOR DISEASE PROBLEMS .................................................................................................... 171
4. DISEASES IN LABORATORY MICE .............................................................................................. 172
5. KEY TAKEAWAYS LABORATORY MOUSE DISEASES............................................................................ 176
6. DISEASES IN LABORATORY RATS ............................................................................................... 176
7. RESPONSE PLAN (FOR ANY DISEASE OUTBREAK) ........................................................................... 180
3
,CH11: RODENTS FIELD RESEARCH (H. LEIRS) .................................................................... 181
1. CAPTURE METHODS ............................................................................................................. 181
2. TRAPPING EFFORT ................................................................................................................ 183
3. MARKING RODENTS .............................................................................................................. 184
4. SAMPLE COLLECTION............................................................................................................ 185
5. HUMANE KILLING ................................................................................................................. 186
6. BIOSAFETY ......................................................................................................................... 187
7. STUDYING BEHAVIOUR IN NATURE ............................................................................................ 188
8. LEGISLATION....................................................................................................................... 189
9. PUBLIC VISIBILITY................................................................................................................. 189
10. ALTERNATIVE METHODS WITHOUT RODENT MANIPULATION ............................................................ 190
4
,CH 0: INTRODUCTION
This summary covers the full theoretical content of the course Laboratory Animal Science (LAS) –
Rodents. All lecture material has been processed and integrated, including detailed notes taken
during the classes. In addition, new chapters that must be studied independently are included, such
as CH11: Rodents in Field Research.
Throughout the summary, exam tips are clearly indicated, and a distinction is made between material
that is essential and content that is of lesser importance.
The exam consists of approximately 10-15 open-ended questions.
Good luck!
5
,CH1: LEGISLATION (C . V A N G IN N IK E N )
1. LEGISLATION ON WORKING WITH LABORATORY ANIMALS
This chapter is revision from the core module (1st master) à not important for the exam!
EUROPEAN LEVEL
• EU Directive 1986
o 1st directive on protection of animals used for experimental & other scientific purposes.
o Required all EU Member States to implement it into their national legislation.
o Belgium implemented it through its Animal Welfare Law (1986) and Royal Decree
(1993).
• EU Directive 2010
o Updated and replaced Directive 1986
o Strengthened protection for animals used in science.
o Key improvements:
§ Clearer rules on the 3Rs (Replacement, Reduction, Refinement).
§ Stricter project authorization requirements.
§ Increased transparency and reporting.
o Required Belgium (and other EU Member States) to update national laws accordingly.
NATIONAL LEVEL (BELGIUM)
• Animal Welfare Law (1986)
o Foundation for all animal welfare legislation in Belgium.
o Protects the welfare of animals in general — not spec. targeted at laboratory animals
• Royal Decree (RD) 1993
o Specifically addressed the protection of laboratory animals.
o Implemented the EU directive 1986
• Royal Decree (RD) 2013
o Replaced the 1993 RD to implement changes from EU Directive 2010
o Includes annexes with detailed husbandry and care requirements (important for
practical assignments)
• Royal Decree (RD) 2001
o Prohibits certain types of animal experiments
§ Tobacco (research).
§ Cosmetics (except in vivo acute toxicity tests).
§ Monoclonal antibodies using the ascites method (exemptions may be possible
6
, • Decrees in 2017
o Changes related to laboratory animal protection.
o Defines the composition of the Vlaamse Proefdierencommissie (Flemish Animal
Experiment Commission).
REGIONAL LEVEL (FLANDERS & WALLONIA)
• 2017 – Competences for welfare became regional matter (Flanders & Wallonia responsible).
• Vlaamse Codex Dierenwelzijn 2024
o New Flemish Animal Welfare Code, eiective 1 January 2025.
o Updates and consolidates the 1986 Animal Welfare Law.
o Important future measures:
§ From 2026: Ban on selling animals at markets.
§ From 2029: Mandatory shelters for animals kept outdoors (e.g., horses).
CONSTITUTIONAL LEVEL (BELGIUM)
• Belgian Constitution – 2024 Amendment
o Oiicially recognizes animals as sentient beings.
o States that their welfare must be protected.
o All laws, decrees, and regulations must comply with this constitutional principle.
SCOPE OF PROTECTION
• Applies to vertebrates (unless specifically excluded).
• Protection focuses on:
o Welfare, health, and proper housing conditions.
o Minimizing pain, suiering, and distress.
o Promoting alternatives (3Rs).
7
,2. LABORATORY ANIMAL – DEFINITION
According to the 1986 Animal Welfare Law, laboratory animals include:
• All vertebrates and cephalopods (e.g., octopuses and squids), as these animals can suier.
• Developmental stage:
o Vertebrates: From the last trimester of normal development onward
§ (e.g., mouse embryos become laboratory animals after 14 days of gestation
since their total pregnancy is 21 days) (know the gestation period of the rodents seen in this course)
§ Specifications for fish: considered independently feeding when both ends of the
intestines are open, and the fish takes in food
§ Specifications for birds: when they hatch or are born alive (whether by human
action or not) they are considered experimental animals
o Vertebrates without gestation (e.g., larval zebrafish): considered a laboratory animal
once it becomes self-feeding (e.g. larval zebrafish starting from 72h are lab animals).
When you start using the animal before the last trimester, however the handling exceeds the third
trimester à the animal IS considered a laboratory animal!!
Royal Decree 2001 (RD2001) – Ban on Great Ape Experiments
• No animal experiments allowed on: Chimpanzees, Bonobos, Orangutans, Gorillas.
• Keep in mind: any kind of handling on great apes à Not an animal experiment à simply illegal!
3. LABORATORY ANIMAL – ORIGIN
PURPOSE-BRED ANIMALS
• Certain species must be purpose-bred for scientific use (come from licensed breeding
companies)
1. Rodents: Mouse, Rat, Guinea Pig, Syrian Hamster, Chinese Hamster, Mongolian Gerbil
2. Larger Mammals: Rabbit, Dog, Cat
3. All species of non-Human Primates (NHPs) (F2 generation!)
§ F1 generation should also be purpose bred
§ Grandparents may be caught in the wild (not common)
4. Other species: Frog, Zebrafish
• Why purpose-bred animals?
o Known genetic, biological, and behavioral background à Increases standardization.
o Improves quality and reliability of results.
o Reduces variability à statistical power à ¯procedures & animal use.
8
, • Exemptions:
o If purpose-bred animals are unavailable, researchers must:
§ Prove insuiicient supply from licensed breeders.
§ Ensure animals were bred in good conditions/circumstances
§ Obtain government approval.
USE OF NON-HUMAN PRIMATES (NHPs)
• NHPs can only be used if absolutely necessary (essential for the benefit of humans) and no
other methods (nor alternative animal species) are yet available.
• Allowed only for:
1. Basic research.
2. Applied research (only for human diseases and disorders).
3. Drug and treatment development (including safety studies).
• Not allowed for:
o Protection or conservation of species.
o Education.
o Forensic research.
USE OF NON-PURPOSE-BRED ANIMALS
• If a species is not listed as purpose-bred (see above), researchers can source them elsewhere,
but should:
o Prefer licensed breeders (ensures good housing, quality, and reduces variability).
o Farm animals are an exception (since most farmers lack a license).
SPECIAL CASES
• Endangered Species
o Generally not allowed, but exemptions exist if:
§ The research directly benefits the species itself.
§ No alternative species or method is available.
§ Used only for basic/applied research, drug development, or species
preservation.
• Wild-Caught Animals
o Allowed only with a government license.
o Researchers must prove no alternative species or method exists.
o Must be caught by a competent person
9
ANIMAL SCIENCE
(LAS) - RODENTS
UAntwerpen
,TABLE OF CONTENTS
CH 0: INTRODUCTION ..............................................................................................................5
CH1: LEGISLATION (C. VAN GINNIKEN) ..................................................................................6
1. LEGISLATION ON WORKING WITH LABORATORY ANIMALS ...................................................................... 6
2. LABORATORY ANIMAL – DEFINITION ............................................................................................... 8
3. LABORATORY ANIMAL – ORIGIN ..................................................................................................... 8
4. LABORATORY ANIMAL – IDENTIFICATION ........................................................................................ 10
5. ANIMAL EXPERIMENT – PURPOSES ............................................................................................... 11
6. ETHICAL COMMITTEE ............................................................................................................... 12
7. ANIMAL WELFARE UNIT/BODY ..................................................................................................... 13
8. USER, SUPPLIER, BREEDER ........................................................................................................ 13
9. ANIMAL HUSBANDRY ............................................................................................................... 14
10. PERSONNEL AND THEIR RESPONSABILITIES (!) ................................................................................. 15
11. AT THE END OF THE EXPERIMENT – OUTCOMES ................................................................................ 17
CH2: PRINCIPLES OF ANIMAL HANDLING AND RESTRAINT (D. VAN DAM) .............................. 19
1. THE 3HS-MODULES ................................................................................................................ 19
2. ANIMAL HANDLING ................................................................................................................. 19
3. NON-AVERSIVE / REFINED HANDLING OF MICE ................................................................................ 21
4. RAT HANDLING / TOUCHING ...................................................................................................... 25
5. PICKING UP ANIMALS ............................................................................................................... 27
6. IMMOBILISATION OF ANIMALS..................................................................................................... 28
CH3: BIOLOGY OF RODENTS (C. VAN GINNIKEN) .................................................................. 33
1. MOUSE – NUMBERS (BELGIUM / FLANDERS) ................................................................................... 33
2. MOUSE – BIOLOGY ................................................................................................................. 33
3. ANIMAL HUSBANDRY CONSIDERATIONS (GENERAL) ........................................................................ 36
4. MOUSE HUSBANDRY............................................................................................................... 37
5. MOUSE – ENVIRONMENTAL PARAMETERS ..................................................................................... 41
6. MOUSE – ANATOMICAL & PHYSIOLOGICAL FEATURES ...................................................................... 42
7. MOUSE – NUTRITION & REPRODUCTION ...................................................................................... 46
8. MOUSE – REPRODUCTIVE AND URINARY SYSTEM ............................................................................. 47
9. MOUSE – FERTILITY CYCLE, BREEDING & REPRODUCTION ................................................................ 48
10. MOUSE – DEVELOPMENT & BREEDING PROTOCOLS ........................................................................ 51
11. RAT – NUMBERS .................................................................................................................... 54
12. RAT – BIOLOGY...................................................................................................................... 54
13. RAT – HUSBANDRY ................................................................................................................. 55
14. RAT – ENVIRONMENTAL PARAMETERS, ANATOMY & REPRODUCTION .................................................... 56
1
,15. RAT – BREEDING & REPRODUCTION ............................................................................................ 57
CH4: ANIMAL WELFARE (D. VAN DAM) .................................................................................. 59
1. IMPACT ON ANIMAL WELFARE IN RESEARCH .................................................................................. 59
2. ANIMAL WELFARE AND SENTIENCE ............................................................................................. 60
3. CONTROLLABILITY AND PREDICTABILITY ........................................................................................ 64
4. FREQUENCY OF STRESSORS & HABITUATION .................................................................................. 66
5. REGULATORY RANGE & ADAPTIVE CAPACITY ................................................................................. 67
6. STRESS, DISTRESS & ANIMAL WELFARE ....................................................................................... 68
7. HABITUATION VS. SENSITIZATION................................................................................................ 69
8. STRESS THROUGHOUT THE LIFE CYCLE (EXAMPLES) ........................................................................ 69
CH5: BEHAVIORAL EVALUATION (VAN DAM) ......................................................................... 75
1. GOAL OF BEHAVIORAL EVALUATION ............................................................................................ 75
2. MULTI-TIERED EVALUATION PROCESS ......................................................................................... 75
3. TIER 1: GENERAL HEALTH AND NEUROLOGICAL REFLEXES (DETAILS) ................................................... 76
4. SENSORY AND MOTOR ABILITIES ................................................................................................ 81
5. SPECIFIC BEHAVIORAL DOMAINS (3RD TIER): SOCIAL BEHAVIORS ....................................................... 87
6. FEAR AND ANXIETY ................................................................................................................. 91
7. CIRCADIAN RHYTHM AND SLEEP ................................................................................................ 92
8. LEARNING AND MEMORY (COGNITIVE FUNCTION TESTS) .................................................................. 93
9. DEPRESSION-RELATED SYMPTOMS ............................................................................................. 97
CH6: MINIMALLY INVASIVE PROCEDURES (C. VAN GINNIKEN) .............................................. 98
1. IDENTIFICATION / MARKING OF ANIMALS ....................................................................................... 98
2. ADMINISTRATION OF SUBSTANCES............................................................................................. 103
3. COLLECTION OF BODY FLUIDS .................................................................................................. 108
CH7: ANESTHESIA AND ANALGESIA (C. VAN GINNEKEN) .................................................... 117
1. PRE-OPERATIVE MANAGEMENT ................................................................................................ 117
2. PRE-OPERATIVE: FASTING ...................................................................................................... 120
3. PERI-OPERATIVE PERIOD – MANAGEMENT................................................................................... 122
4. DEFINITIONS ....................................................................................................................... 123
5. ANESTHESIA ....................................................................................................................... 124
6. THE TRIAD OF ANESTHESIA ..................................................................................................... 124
7. ANESTHESIA – PHASES .......................................................................................................... 125
8. ANESTHETIC DEPTH .............................................................................................................. 125
9. INHALATION ANESTHETICS...................................................................................................... 127
10. INJECTABLE ANAESTHETICS ..................................................................................................... 132
11. ANAESTHESIA – PHARMACA .................................................................................................... 137
2
,12. OBSERVATIONAL MONITORING DURING ANAESTHESIA ..................................................................... 138
13. TROUBLESHOOTING .............................................................................................................. 139
14. POST-OPERATIVE MANAGEMENT ............................................................................................... 140
15. PAIN & ANALGESIA ............................................................................................................... 141
16. PRE-EMPTIVE ANALGESIA ....................................................................................................... 141
17. PAIN MANAGEMENT – ANALGESICS ........................................................................................... 142
18. PAIN ASSESSMENT ............................................................................................................... 144
CH8: PAIN AND WELFARE ASSESSMENT (D. VAN DAM) ....................................................... 146
1. PAIN AND WELFARE ASSESSMENT ............................................................................................ 146
2. WELFARE ASSESSMENT ......................................................................................................... 146
3. WELFARE INDICATORS........................................................................................................... 146
4. TUTORIAL: ANIMAL HEALTH AND WELFARE (AHWLA) ................................................................... 147
5. BODY CONDITION ................................................................................................................ 147
6. BODY SHAPE AND MALFORMATIONS ......................................................................................... 148
7. POSTURE ........................................................................................................................... 149
8. FUR QUALITY ...................................................................................................................... 150
9. EYES AND MOUTH ................................................................................................................ 150
10. SKIN AND MUCOUS MEMBRANES ............................................................................................. 151
11. TAIL .................................................................................................................................. 152
12. BEHAVIOURAL ASPECTS ......................................................................................................... 152
13. PAIN ASSESSMENT ............................................................................................................... 154
14. GRIMACE SCALE (FACIAL EXPRESSION ANALYSIS)......................................................................... 159
15. ESSENTIAL KNOWLEDGE FOR WELFARE ASSESSMENT .................................................................... 161
16. WELFARE ASSESSMENT IN GENETICALLY MODIFIED (GM) RODENTS .................................................. 162
CH9: HUMANE KILLING ........................................................................................................ 164
1. LEGISLATION ON HUMANE KILLING ........................................................................................... 164
2. REQUIREMENTS FOR HUMANE KILLING ...................................................................................... 164
3. METHODS FOR HUMANE KILLING (EXAM) ................................................................................. 164
4. CONFIRMATION OF DEATH...................................................................................................... 167
5. HUMANE ENDPOINTS ............................................................................................................ 167
CH10: DISEASES (C. VAN GINNEKEN) ............................................................................... 168
1. HOW MICROORGANISMS ENTER THE ANIMAL UNIT ......................................................................... 168
2. HEALTH MONITORING (HM) ................................................................................................... 169
3. MAJOR DISEASE PROBLEMS .................................................................................................... 171
4. DISEASES IN LABORATORY MICE .............................................................................................. 172
5. KEY TAKEAWAYS LABORATORY MOUSE DISEASES............................................................................ 176
6. DISEASES IN LABORATORY RATS ............................................................................................... 176
7. RESPONSE PLAN (FOR ANY DISEASE OUTBREAK) ........................................................................... 180
3
,CH11: RODENTS FIELD RESEARCH (H. LEIRS) .................................................................... 181
1. CAPTURE METHODS ............................................................................................................. 181
2. TRAPPING EFFORT ................................................................................................................ 183
3. MARKING RODENTS .............................................................................................................. 184
4. SAMPLE COLLECTION............................................................................................................ 185
5. HUMANE KILLING ................................................................................................................. 186
6. BIOSAFETY ......................................................................................................................... 187
7. STUDYING BEHAVIOUR IN NATURE ............................................................................................ 188
8. LEGISLATION....................................................................................................................... 189
9. PUBLIC VISIBILITY................................................................................................................. 189
10. ALTERNATIVE METHODS WITHOUT RODENT MANIPULATION ............................................................ 190
4
,CH 0: INTRODUCTION
This summary covers the full theoretical content of the course Laboratory Animal Science (LAS) –
Rodents. All lecture material has been processed and integrated, including detailed notes taken
during the classes. In addition, new chapters that must be studied independently are included, such
as CH11: Rodents in Field Research.
Throughout the summary, exam tips are clearly indicated, and a distinction is made between material
that is essential and content that is of lesser importance.
The exam consists of approximately 10-15 open-ended questions.
Good luck!
5
,CH1: LEGISLATION (C . V A N G IN N IK E N )
1. LEGISLATION ON WORKING WITH LABORATORY ANIMALS
This chapter is revision from the core module (1st master) à not important for the exam!
EUROPEAN LEVEL
• EU Directive 1986
o 1st directive on protection of animals used for experimental & other scientific purposes.
o Required all EU Member States to implement it into their national legislation.
o Belgium implemented it through its Animal Welfare Law (1986) and Royal Decree
(1993).
• EU Directive 2010
o Updated and replaced Directive 1986
o Strengthened protection for animals used in science.
o Key improvements:
§ Clearer rules on the 3Rs (Replacement, Reduction, Refinement).
§ Stricter project authorization requirements.
§ Increased transparency and reporting.
o Required Belgium (and other EU Member States) to update national laws accordingly.
NATIONAL LEVEL (BELGIUM)
• Animal Welfare Law (1986)
o Foundation for all animal welfare legislation in Belgium.
o Protects the welfare of animals in general — not spec. targeted at laboratory animals
• Royal Decree (RD) 1993
o Specifically addressed the protection of laboratory animals.
o Implemented the EU directive 1986
• Royal Decree (RD) 2013
o Replaced the 1993 RD to implement changes from EU Directive 2010
o Includes annexes with detailed husbandry and care requirements (important for
practical assignments)
• Royal Decree (RD) 2001
o Prohibits certain types of animal experiments
§ Tobacco (research).
§ Cosmetics (except in vivo acute toxicity tests).
§ Monoclonal antibodies using the ascites method (exemptions may be possible
6
, • Decrees in 2017
o Changes related to laboratory animal protection.
o Defines the composition of the Vlaamse Proefdierencommissie (Flemish Animal
Experiment Commission).
REGIONAL LEVEL (FLANDERS & WALLONIA)
• 2017 – Competences for welfare became regional matter (Flanders & Wallonia responsible).
• Vlaamse Codex Dierenwelzijn 2024
o New Flemish Animal Welfare Code, eiective 1 January 2025.
o Updates and consolidates the 1986 Animal Welfare Law.
o Important future measures:
§ From 2026: Ban on selling animals at markets.
§ From 2029: Mandatory shelters for animals kept outdoors (e.g., horses).
CONSTITUTIONAL LEVEL (BELGIUM)
• Belgian Constitution – 2024 Amendment
o Oiicially recognizes animals as sentient beings.
o States that their welfare must be protected.
o All laws, decrees, and regulations must comply with this constitutional principle.
SCOPE OF PROTECTION
• Applies to vertebrates (unless specifically excluded).
• Protection focuses on:
o Welfare, health, and proper housing conditions.
o Minimizing pain, suiering, and distress.
o Promoting alternatives (3Rs).
7
,2. LABORATORY ANIMAL – DEFINITION
According to the 1986 Animal Welfare Law, laboratory animals include:
• All vertebrates and cephalopods (e.g., octopuses and squids), as these animals can suier.
• Developmental stage:
o Vertebrates: From the last trimester of normal development onward
§ (e.g., mouse embryos become laboratory animals after 14 days of gestation
since their total pregnancy is 21 days) (know the gestation period of the rodents seen in this course)
§ Specifications for fish: considered independently feeding when both ends of the
intestines are open, and the fish takes in food
§ Specifications for birds: when they hatch or are born alive (whether by human
action or not) they are considered experimental animals
o Vertebrates without gestation (e.g., larval zebrafish): considered a laboratory animal
once it becomes self-feeding (e.g. larval zebrafish starting from 72h are lab animals).
When you start using the animal before the last trimester, however the handling exceeds the third
trimester à the animal IS considered a laboratory animal!!
Royal Decree 2001 (RD2001) – Ban on Great Ape Experiments
• No animal experiments allowed on: Chimpanzees, Bonobos, Orangutans, Gorillas.
• Keep in mind: any kind of handling on great apes à Not an animal experiment à simply illegal!
3. LABORATORY ANIMAL – ORIGIN
PURPOSE-BRED ANIMALS
• Certain species must be purpose-bred for scientific use (come from licensed breeding
companies)
1. Rodents: Mouse, Rat, Guinea Pig, Syrian Hamster, Chinese Hamster, Mongolian Gerbil
2. Larger Mammals: Rabbit, Dog, Cat
3. All species of non-Human Primates (NHPs) (F2 generation!)
§ F1 generation should also be purpose bred
§ Grandparents may be caught in the wild (not common)
4. Other species: Frog, Zebrafish
• Why purpose-bred animals?
o Known genetic, biological, and behavioral background à Increases standardization.
o Improves quality and reliability of results.
o Reduces variability à statistical power à ¯procedures & animal use.
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, • Exemptions:
o If purpose-bred animals are unavailable, researchers must:
§ Prove insuiicient supply from licensed breeders.
§ Ensure animals were bred in good conditions/circumstances
§ Obtain government approval.
USE OF NON-HUMAN PRIMATES (NHPs)
• NHPs can only be used if absolutely necessary (essential for the benefit of humans) and no
other methods (nor alternative animal species) are yet available.
• Allowed only for:
1. Basic research.
2. Applied research (only for human diseases and disorders).
3. Drug and treatment development (including safety studies).
• Not allowed for:
o Protection or conservation of species.
o Education.
o Forensic research.
USE OF NON-PURPOSE-BRED ANIMALS
• If a species is not listed as purpose-bred (see above), researchers can source them elsewhere,
but should:
o Prefer licensed breeders (ensures good housing, quality, and reduces variability).
o Farm animals are an exception (since most farmers lack a license).
SPECIAL CASES
• Endangered Species
o Generally not allowed, but exemptions exist if:
§ The research directly benefits the species itself.
§ No alternative species or method is available.
§ Used only for basic/applied research, drug development, or species
preservation.
• Wild-Caught Animals
o Allowed only with a government license.
o Researchers must prove no alternative species or method exists.
o Must be caught by a competent person
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