LABORATORY ANIMAL SCIENCE: FUNCTION B
TABLE OF CONTENTS
1. Lab animals in Flanders - an historical and ethical perspective ...................................................................................................................... 1
Prehistorical times ........................................................................................................................................................................................ 1
Middle Ages .................................................................................................................................................................................................. 1
Renaissance (14-16th century) ...................................................................................................................................................................... 1
Age of Enlightenment ................................................................................................................................................................................... 2
Modern times (1750-1950) .......................................................................................................................................................................... 3
In Belgium..................................................................................................................................................................................................... 5
Ethics ............................................................................................................................................................................................................ 7
2.1 Legislation part 1.......................................................................................................................................................................................... 8
European level .............................................................................................................................................................................................. 8
National level (in Belgium) ........................................................................................................................................................................... 8
Laboratory animal ........................................................................................................................................................................................ 8
Animal experiment ..................................................................................................................................................................................... 11
Ethical committee ...................................................................................................................................................................................... 13
User, breeder, supplier ............................................................................................................................................................................... 14
Animal husbandry....................................................................................................................................................................................... 14
Personnel and their responsabilities .......................................................................................................................................................... 15
At the end of the experiment? ................................................................................................................................................................... 17
2.2 Legislation part 2........................................................................................................................................................................................ 20
Ethics committee ........................................................................................................................................................................................ 20
Animal welfare body .................................................................................................................................................................................. 24
National committee.................................................................................................................................................................................... 25
3.1 Numbers and species ................................................................................................................................................................................. 26
Animal welfare ........................................................................................................................................................................................... 26
Numbers of animals used ........................................................................................................................................................................... 26
Biological characteristics of the most frequently used species................................................................................................................... 28
3.2 Animal husbandry incl. safety .................................................................................................................................................................... 36
Husbandry (housing) .................................................................................................................................................................................. 36
Nutrition ..................................................................................................................................................................................................... 40
Transport .................................................................................................................................................................................................... 42
3.3 safety incl. health monitoring .................................................................................................................................................................... 43
Health monitoring ...................................................................................................................................................................................... 43
Risks............................................................................................................................................................................................................ 45
4. Animal welfare: general principles ............................................................................................................................................................... 47
Sentient beings ........................................................................................................................................................................................... 47
Animal welfare ........................................................................................................................................................................................... 47
Learning processes ..................................................................................................................................................................................... 48
Concept of five freedoms ........................................................................................................................................................................... 49
Pain and welfare assessment ..................................................................................................................................................................... 53
5.1 Project design part 1 .................................................................................................................................................................................. 56
PREPARE guidelines .................................................................................................................................................................................... 56
, Basics for good experimental design .......................................................................................................................................................... 58
Statistical methods to support experimental design .................................................................................................................................. 61
Experimental designs.................................................................................................................................................................................. 63
ARRIVE guidelines ....................................................................................................................................................................................... 67
5.2 Experimental design part 2 ........................................................................................................................................................................ 69
Alternatives for animal-based research ...................................................................................................................................................... 69
Severity assessment ................................................................................................................................................................................... 71
Humane endpoints ..................................................................................................................................................................................... 74
6. Minimally invasive procedures..................................................................................................................................................................... 77
Marking of animaLs .................................................................................................................................................................................... 77
Administration of substances ..................................................................................................................................................................... 79
Collection of body fluids ............................................................................................................................................................................. 81
7.1 Asepsis ....................................................................................................................................................................................................... 84
Halsteds principles...................................................................................................................................................................................... 84
Basic surgery............................................................................................................................................................................................... 84
Aseptic techniques ..................................................................................................................................................................................... 85
7.2 Peri-operative management and care........................................................................................................................................................ 89
Pre-operative management........................................................................................................................................................................ 89
Post-operative management ...................................................................................................................................................................... 90
7.3 Suturing and wound healing ...................................................................................................................................................................... 92
Suture material (cf. practical) ..................................................................................................................................................................... 92
Suture techniques (cf. practical) ................................................................................................................................................................. 94
Wound healing ........................................................................................................................................................................................... 95
8.1 Anesthesia.................................................................................................................................................................................................. 97
Key elements .............................................................................................................................................................................................. 97
Phases ........................................................................................................................................................................................................ 98
Most frequently used anesthetics .............................................................................................................................................................. 99
Possible complications and how to respond ............................................................................................................................................ 103
Examples .................................................................................................................................................................................................. 104
8.2 Analgesia .................................................................................................................................................................................................. 106
Pain management .................................................................................................................................................................................... 106
Most frequently used analgesics .............................................................................................................................................................. 107
Pain assessment ....................................................................................................................................................................................... 109
9. Humane killing and humane endpoints ..................................................................................................................................................... 110
Humane killing .......................................................................................................................................................................................... 110
Humane endpoints ................................................................................................................................................................................... 114
10. Genetic modification ................................................................................................................................................................................ 115
Selective breeding .................................................................................................................................................................................... 115
Inbred vs. outbred strains ........................................................................................................................................................................ 115
Spontaneous vs. induced models ............................................................................................................................................................. 118
Genetic modification revolution ............................................................................................................................................................... 121
,Opinions about laboratory animals and animal experiments vary widely. But what actually are a laboratory animal
and an animal experiment? This is still often misunderstood by general population!
1. LAB ANIMALS IN FLANDERS - AN HISTORICAL AND ETHICAL PERSPECTIVE
PREHISTORICAL TIMES
Aristotle (384-322 BC)
• Scala naturae (natural order)
o From simple, low level of importance to more complex, nearly perfect, high level of importance
o = introduction to the thinking of higher and lower beings; we still do this cf. lower vertebrates
Plutarchus (46-120) did not agree with this thinking
• Involves animals in ethical considerations
• Vegetarism
• Pythagoras
Galenus (129-216)
• Taboo on using human bodies (but Roman Games)
• Vivisection and dissection
o Vivisection: surgery conducted for experimental purposes on a living organism without anesthesia
• Uses monkeys and pigs as proxy for the human body
• His knowledge prevailed until 17th century
MIDDLE AGES
Augustinus Hippo (354-430)
• Scala naturae added with angels and God
o In order for men to get closer to God, distance between
men and animals increased
o Forsake animal instincts
• Man to Gods image
• Animals serve man
No real progress: disease = curse and cure = faith
RENAISSANCE (14-16 T H CENTURY)
= revival and critical view on the ancient times
Vesalius (1514-1564): corrected Aristotle and Galenus by using human bodies
Van Leeuwen: invention of the microscope
→ Different view on the position of men; however, vivisection of animals was still not seen as a problem
→ Start of a more scientific approach
1
,AGE OF ENLIGHTENMENT
Descartes (1596-1650)
• Theory of innate knowledge: all humans were born with knowledge through the higher power of God
o ↔ John Locke (1632-1704), an empiricist: all knowledge is acquired through experience
• ‘Je pense donc je suis’
• Importance of reasoning
• Mens innate knowledge and capability to reason, think, feel… differentiate them from animals, which are
merely machines (automata) with which we can do what we want
Industrialization
• Men became less acquainted with nature; decreasing contact with animals made it even more difficult to
have feel for animal welfare
• Men had more time to think about the principles of life
Hogarth (1751)
• Four stages of Cruelty
o Show the story of Tom Nero in an easy to understand way (many people could not read back then)
o Point of the story: if you mistreat animals, the chance of you mistreating other humans is also bigger
BUT remember that the same cruel things can be done to you!
o First time that this type of considered thinking about the handling of animals was published
2
,Kant (1724-1804): deontology
• Actions that we must take (duty) are based on reasoning and do not consider the outcome or consequences
• Categorical imperative
o Treat human(ity) as a goal, never as a means
o Universal (for all and under all circumstances)
• Reasoning is the key that decides what has dignity and what not → only (hu)man
• ‘Animals are there merely as a means to an end. That end is man.’
• ‘We can judge the heart of a man by his treatment of animals.’
Bentham (1748-1832) and Mill: utilism
• Agrees to the use of animals as long as the benefit exceeds the suffering for man
• → Calculus
o Everybody is to count for one, nobody to count for more than one
• Greatest happiness principle
o Reasoning is less important than happiness
o Considers the consequences of actions (↔ Kant)
• ‘The question is not Can they reason, nor Can they talk, but Can they suffer…‘→ sentient beings
o This lies at the basis of the 3R principle
MODERN TIMES (1750-1950)
Charles Darwin
• Origin of species > scala naturae
• Start of zoocentrism: ethical theories that confer moral standing towards at least some animals
Claude Bernard (humanist; 1813-1878)
• Foundation of animal experiments, which should be properly controlled
• First test in animals
Cruelty against animals act (1867; UK)
• Cruel treatment of cattle act (1822)
• Cruelty against animals act (1835)
o Reaction against Magendie (France), who sold tickets to his vivisections
• Rules concerning animal research
o Prosecution is possible
o Only vertebrates
o Under anesthesia
o Use each animal only 1x
o Immediately kill animal after the experiment
o Only when necessary to advance human health
→ UK was the first country that included in their legislation that you cannot be cruel to animals
3
, Even though people were realizing that animal experiments are not harmless thanks to advent of psychology
(Wolfang Köhler, Jane Goodall, Frans De Waal), there was a rise in animal experiments because of advancing
science
• Infections
o Kochs postulates
o Pasteur used humane endpoints; he killed the animals once they had suffered enough and were
not going to recover anymore
• Scientifically tested drugs, vaccines,…
• Use of rodents (= pest animals)
→ Difficult for anti-vivisectionists (National Anti-vivisection Society) to upheld the arguments that no
advancement is made via animal experiments → discussion shifted to preventing harm rather than questioning
the value of animal research
Anesthetics (1846)
• Gave carte blanche (you could do whatever you wanted with anesthetized animal) → increase in animal use
• Often failed to work
Russel and Burch (1959; UK)
• “The Principles of Humane Experimental Technique” (re-emerged in 1987)
o ‘We assume that experimental biologists are only too happy to treat their animals as humanely as
possible. BUT The central problem then is that of determining what is and what is not humane, and
how humanity can be promoted without prejudice to scientific and medical aims. We must begin
by examining the concept of humanity (or inhumanity) as an objective assessment of the effects of
any procedure on the animal subject.’
o → 3Rs: if there is no suitable alternative (replacement), use the lowest number of animals possible
(reduction) and keep their lives as good as possible (refinement)
Replacement Reduction Refinement
• Non-vertebrate model • Sample size calculations • Cage enrichment
• Embryonic/larval form • Re-use • Use of anesthetics and analgesics
• In vitro test • Reporting negative data • In vivo imaging
• Computer/in silico
Tom Regan (US)
• “The Case for Animal Rights” (1983)
o Extension of Kants deontology
o Animal rights (≠ animal welfare)
• Moral agents vs. moral patients
o Moral agents are those who have full autonomy and are capable (possess the necessary cognitive
skills) in making decisions
o Moral patients are individuals who have desires, longings, know emotions,…
o Only normal adult humans are moral agents, all others are subjects of life and thus moral patients
who have fundamental (universal) rights
o Moral agents have the duty to respect the rights of moral patients
• All subjects of life should be considered as a goal, not as a means
• Opposes all use of animals; no animal experiments, no eating of animals, no pet animals,…
4
TABLE OF CONTENTS
1. Lab animals in Flanders - an historical and ethical perspective ...................................................................................................................... 1
Prehistorical times ........................................................................................................................................................................................ 1
Middle Ages .................................................................................................................................................................................................. 1
Renaissance (14-16th century) ...................................................................................................................................................................... 1
Age of Enlightenment ................................................................................................................................................................................... 2
Modern times (1750-1950) .......................................................................................................................................................................... 3
In Belgium..................................................................................................................................................................................................... 5
Ethics ............................................................................................................................................................................................................ 7
2.1 Legislation part 1.......................................................................................................................................................................................... 8
European level .............................................................................................................................................................................................. 8
National level (in Belgium) ........................................................................................................................................................................... 8
Laboratory animal ........................................................................................................................................................................................ 8
Animal experiment ..................................................................................................................................................................................... 11
Ethical committee ...................................................................................................................................................................................... 13
User, breeder, supplier ............................................................................................................................................................................... 14
Animal husbandry....................................................................................................................................................................................... 14
Personnel and their responsabilities .......................................................................................................................................................... 15
At the end of the experiment? ................................................................................................................................................................... 17
2.2 Legislation part 2........................................................................................................................................................................................ 20
Ethics committee ........................................................................................................................................................................................ 20
Animal welfare body .................................................................................................................................................................................. 24
National committee.................................................................................................................................................................................... 25
3.1 Numbers and species ................................................................................................................................................................................. 26
Animal welfare ........................................................................................................................................................................................... 26
Numbers of animals used ........................................................................................................................................................................... 26
Biological characteristics of the most frequently used species................................................................................................................... 28
3.2 Animal husbandry incl. safety .................................................................................................................................................................... 36
Husbandry (housing) .................................................................................................................................................................................. 36
Nutrition ..................................................................................................................................................................................................... 40
Transport .................................................................................................................................................................................................... 42
3.3 safety incl. health monitoring .................................................................................................................................................................... 43
Health monitoring ...................................................................................................................................................................................... 43
Risks............................................................................................................................................................................................................ 45
4. Animal welfare: general principles ............................................................................................................................................................... 47
Sentient beings ........................................................................................................................................................................................... 47
Animal welfare ........................................................................................................................................................................................... 47
Learning processes ..................................................................................................................................................................................... 48
Concept of five freedoms ........................................................................................................................................................................... 49
Pain and welfare assessment ..................................................................................................................................................................... 53
5.1 Project design part 1 .................................................................................................................................................................................. 56
PREPARE guidelines .................................................................................................................................................................................... 56
, Basics for good experimental design .......................................................................................................................................................... 58
Statistical methods to support experimental design .................................................................................................................................. 61
Experimental designs.................................................................................................................................................................................. 63
ARRIVE guidelines ....................................................................................................................................................................................... 67
5.2 Experimental design part 2 ........................................................................................................................................................................ 69
Alternatives for animal-based research ...................................................................................................................................................... 69
Severity assessment ................................................................................................................................................................................... 71
Humane endpoints ..................................................................................................................................................................................... 74
6. Minimally invasive procedures..................................................................................................................................................................... 77
Marking of animaLs .................................................................................................................................................................................... 77
Administration of substances ..................................................................................................................................................................... 79
Collection of body fluids ............................................................................................................................................................................. 81
7.1 Asepsis ....................................................................................................................................................................................................... 84
Halsteds principles...................................................................................................................................................................................... 84
Basic surgery............................................................................................................................................................................................... 84
Aseptic techniques ..................................................................................................................................................................................... 85
7.2 Peri-operative management and care........................................................................................................................................................ 89
Pre-operative management........................................................................................................................................................................ 89
Post-operative management ...................................................................................................................................................................... 90
7.3 Suturing and wound healing ...................................................................................................................................................................... 92
Suture material (cf. practical) ..................................................................................................................................................................... 92
Suture techniques (cf. practical) ................................................................................................................................................................. 94
Wound healing ........................................................................................................................................................................................... 95
8.1 Anesthesia.................................................................................................................................................................................................. 97
Key elements .............................................................................................................................................................................................. 97
Phases ........................................................................................................................................................................................................ 98
Most frequently used anesthetics .............................................................................................................................................................. 99
Possible complications and how to respond ............................................................................................................................................ 103
Examples .................................................................................................................................................................................................. 104
8.2 Analgesia .................................................................................................................................................................................................. 106
Pain management .................................................................................................................................................................................... 106
Most frequently used analgesics .............................................................................................................................................................. 107
Pain assessment ....................................................................................................................................................................................... 109
9. Humane killing and humane endpoints ..................................................................................................................................................... 110
Humane killing .......................................................................................................................................................................................... 110
Humane endpoints ................................................................................................................................................................................... 114
10. Genetic modification ................................................................................................................................................................................ 115
Selective breeding .................................................................................................................................................................................... 115
Inbred vs. outbred strains ........................................................................................................................................................................ 115
Spontaneous vs. induced models ............................................................................................................................................................. 118
Genetic modification revolution ............................................................................................................................................................... 121
,Opinions about laboratory animals and animal experiments vary widely. But what actually are a laboratory animal
and an animal experiment? This is still often misunderstood by general population!
1. LAB ANIMALS IN FLANDERS - AN HISTORICAL AND ETHICAL PERSPECTIVE
PREHISTORICAL TIMES
Aristotle (384-322 BC)
• Scala naturae (natural order)
o From simple, low level of importance to more complex, nearly perfect, high level of importance
o = introduction to the thinking of higher and lower beings; we still do this cf. lower vertebrates
Plutarchus (46-120) did not agree with this thinking
• Involves animals in ethical considerations
• Vegetarism
• Pythagoras
Galenus (129-216)
• Taboo on using human bodies (but Roman Games)
• Vivisection and dissection
o Vivisection: surgery conducted for experimental purposes on a living organism without anesthesia
• Uses monkeys and pigs as proxy for the human body
• His knowledge prevailed until 17th century
MIDDLE AGES
Augustinus Hippo (354-430)
• Scala naturae added with angels and God
o In order for men to get closer to God, distance between
men and animals increased
o Forsake animal instincts
• Man to Gods image
• Animals serve man
No real progress: disease = curse and cure = faith
RENAISSANCE (14-16 T H CENTURY)
= revival and critical view on the ancient times
Vesalius (1514-1564): corrected Aristotle and Galenus by using human bodies
Van Leeuwen: invention of the microscope
→ Different view on the position of men; however, vivisection of animals was still not seen as a problem
→ Start of a more scientific approach
1
,AGE OF ENLIGHTENMENT
Descartes (1596-1650)
• Theory of innate knowledge: all humans were born with knowledge through the higher power of God
o ↔ John Locke (1632-1704), an empiricist: all knowledge is acquired through experience
• ‘Je pense donc je suis’
• Importance of reasoning
• Mens innate knowledge and capability to reason, think, feel… differentiate them from animals, which are
merely machines (automata) with which we can do what we want
Industrialization
• Men became less acquainted with nature; decreasing contact with animals made it even more difficult to
have feel for animal welfare
• Men had more time to think about the principles of life
Hogarth (1751)
• Four stages of Cruelty
o Show the story of Tom Nero in an easy to understand way (many people could not read back then)
o Point of the story: if you mistreat animals, the chance of you mistreating other humans is also bigger
BUT remember that the same cruel things can be done to you!
o First time that this type of considered thinking about the handling of animals was published
2
,Kant (1724-1804): deontology
• Actions that we must take (duty) are based on reasoning and do not consider the outcome or consequences
• Categorical imperative
o Treat human(ity) as a goal, never as a means
o Universal (for all and under all circumstances)
• Reasoning is the key that decides what has dignity and what not → only (hu)man
• ‘Animals are there merely as a means to an end. That end is man.’
• ‘We can judge the heart of a man by his treatment of animals.’
Bentham (1748-1832) and Mill: utilism
• Agrees to the use of animals as long as the benefit exceeds the suffering for man
• → Calculus
o Everybody is to count for one, nobody to count for more than one
• Greatest happiness principle
o Reasoning is less important than happiness
o Considers the consequences of actions (↔ Kant)
• ‘The question is not Can they reason, nor Can they talk, but Can they suffer…‘→ sentient beings
o This lies at the basis of the 3R principle
MODERN TIMES (1750-1950)
Charles Darwin
• Origin of species > scala naturae
• Start of zoocentrism: ethical theories that confer moral standing towards at least some animals
Claude Bernard (humanist; 1813-1878)
• Foundation of animal experiments, which should be properly controlled
• First test in animals
Cruelty against animals act (1867; UK)
• Cruel treatment of cattle act (1822)
• Cruelty against animals act (1835)
o Reaction against Magendie (France), who sold tickets to his vivisections
• Rules concerning animal research
o Prosecution is possible
o Only vertebrates
o Under anesthesia
o Use each animal only 1x
o Immediately kill animal after the experiment
o Only when necessary to advance human health
→ UK was the first country that included in their legislation that you cannot be cruel to animals
3
, Even though people were realizing that animal experiments are not harmless thanks to advent of psychology
(Wolfang Köhler, Jane Goodall, Frans De Waal), there was a rise in animal experiments because of advancing
science
• Infections
o Kochs postulates
o Pasteur used humane endpoints; he killed the animals once they had suffered enough and were
not going to recover anymore
• Scientifically tested drugs, vaccines,…
• Use of rodents (= pest animals)
→ Difficult for anti-vivisectionists (National Anti-vivisection Society) to upheld the arguments that no
advancement is made via animal experiments → discussion shifted to preventing harm rather than questioning
the value of animal research
Anesthetics (1846)
• Gave carte blanche (you could do whatever you wanted with anesthetized animal) → increase in animal use
• Often failed to work
Russel and Burch (1959; UK)
• “The Principles of Humane Experimental Technique” (re-emerged in 1987)
o ‘We assume that experimental biologists are only too happy to treat their animals as humanely as
possible. BUT The central problem then is that of determining what is and what is not humane, and
how humanity can be promoted without prejudice to scientific and medical aims. We must begin
by examining the concept of humanity (or inhumanity) as an objective assessment of the effects of
any procedure on the animal subject.’
o → 3Rs: if there is no suitable alternative (replacement), use the lowest number of animals possible
(reduction) and keep their lives as good as possible (refinement)
Replacement Reduction Refinement
• Non-vertebrate model • Sample size calculations • Cage enrichment
• Embryonic/larval form • Re-use • Use of anesthetics and analgesics
• In vitro test • Reporting negative data • In vivo imaging
• Computer/in silico
Tom Regan (US)
• “The Case for Animal Rights” (1983)
o Extension of Kants deontology
o Animal rights (≠ animal welfare)
• Moral agents vs. moral patients
o Moral agents are those who have full autonomy and are capable (possess the necessary cognitive
skills) in making decisions
o Moral patients are individuals who have desires, longings, know emotions,…
o Only normal adult humans are moral agents, all others are subjects of life and thus moral patients
who have fundamental (universal) rights
o Moral agents have the duty to respect the rights of moral patients
• All subjects of life should be considered as a goal, not as a means
• Opposes all use of animals; no animal experiments, no eating of animals, no pet animals,…
4
