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Full summary Laboratory Animal Science

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Laboratory Animal Science

Institution
Universiteit Antwerpen (UA)

This is a full summary given in the first masters biomedical sciences at the university of Antwerp. It includes information of the slides, personal notes and extra information. Everything that is covered during class is included in the summary. The different chapters are: 1) introduction, 2) legisl...

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Uploaded on April 14, 2024
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Type Summary

laboratory animals
legislation
animal welfare
project design
animal care
husbandry
animal facility design
genetic modification
anaesthesia and analgesia
surgical techniqu
minimally invasive procedures

Institution
Universiteit Antwerpen (UA)
Education
Biomedische Wetenschappen
Course
Laboratory Animal Science

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Laboratory Animal Science – Core
Chapter 1: introduction

1. History of laboratory animals
In prehistorical times people used animals to look at nature:
- Aristotle (384-322 BC)
o Scala naturae: he started at a low level and ranked all other material
considering himself as a human being. Ex: lower and higher vertebrates
o Introduction of the thinking of higher and lower beings
- But – Plutarchus (46-120)
o He was a teacher of Pythagoras
o He was vegetarian and looked at animals being equal as human beings
o Involved animals in ethical considerations
- Galenus (129-216)
o He saw resemblances between pigs and human beings
o In this time there was also a taboo using human bodies, so they needed
to turn to animals to learn things.
§ They used monkeys and pigs
o Vivisection and dissection
o This knowledge prevailed until 17th century

Now we come to the Middle Ages, then there was a standstill in scientiﬁc progress,
because there were nay other challenges people faced:
- Augustinus Hippo (354-430)
o Natural order of things: God stands on the top, followed by Angels and
Saints, people, birds, ﬁsh, land animals,…. So there was a religious input
o People tried to resemble God as much as possible. They did that by
making a larger distinction of the lower level of animals.
§ Animal instincts were considered not good
§ Animals serve man
- No real progress
o Disease = curse
o Cure = faith

Then we come to the Renaissance. They looked back on the times before, they
rebirthed the way science was practiced at that time:
- Vesalius (1514-1564)
o Corrected Aristotle and Galenus by using human bodies.
o He started looking at bodies and appreciating di[erences between
human and animal bodies
- Van Leeuwen
o Invention of the microscope

, - Di[erent view on the position of man
- More scientiﬁc approach

To the Age of Enlightenment. The ability to reason:
- Descartes (1596-1650)
o Theory of innate knowledge: all humans were born with knowledge
through the higher power of God
§ ßà combated by philosopher John Locke, an empiricist: all
knowledge is acquired through experience
o The importance of reasoning: animals have their own speak but we don’t
understand them, so animals cannot think and speak as we and don’t
understand things like humans do.
o They made a clear distinction between humans, because they are able to
reason and to think and the rest is seen as machines, and they could use
them for whatever things…
o “Je pense donc je suis”
- Halen
- Industrialization
o People were replaced by machines so people had more time to think
o Less contact with animals
- Hogarth (1751)
o He made 4 stages of Cruelty based on Nero’s life
§ As a young boy Nero played with animals in a cruel way
§ He grow older and was a driver and still showed animal cruelty
§ He has a girlfriend. He asked his girlfriend to steel from her boss
but he kept it for himself and he kills her and crosses the border.
So he is also cruel to humans.
§ Then he was taken to the police: all the cruelty he had done was
now done to him.
o Message: if you are cruel to animals at a young age, you are going to be
criminal at later age. So you have to be respectful
- Kant (1724-1804)
o Deontology = ethical theory that uses rules to distinguish right from
wrong.
o He believed that ethical actions follow universal moral laws, such as
“Don't lie. Don't steal. Don't cheat.”
o Actions that we must take (= duties) based on reasoning and not
considering the outcome or consequences.
§ Because you can reason you have the responsibility to others that
cannot reason!
§ Still allow animals for own purposes but you have responsibility to
take care of them!
o Categorical imperative
§ Universal (for all and under all circumstances)
§ Treat human(ity) as a goal, never as a means

, - Bentham (1748-1832) and Mill
o Utilism = a theory of morality that advocates actions that foster happiness
and oppose actions that cause unhappiness. It promotes "the greatest
amount of good for the greatest number of people."
§ Reasoning is less important than happiness
§ Considers the consequences of actions (<-> Kant)
o They discarded Descartes, they said when you can see a subject su[ering
you have to take it into consideration. You have to remain happiness as
long as possible.
o Sentient beings!!!
§ ‘The question is not Can they reason, nor Can they talk, but Can
they su[er …’
o Calculus
§ Everybody to count for one, nobody to count for more than one.
o Agrees to the use of animals as long as the su[ering is exceeded by the
beneﬁt for man!

In modern times…:
- Charles Darwin (1809-1882)
o We are related to animals, revolutionary view
o Origin of species > Scala naturae
o Start of zoocentrism
- Claude Bernard (1838-1878)
o Assistant of Magendie
o Further elaborating on how to set up an animal experiment
o Foundation of animal experiments (should be properly controlled) – ﬁrst
test in animals (humanist)
- Anesthesia (1846)

In the UK: Cruelty against animals act (1867)
- Cruel treatment of Cattle Act (°1822)
- Cruelty against Against Animals Act (°1835)
- First legislation where animal research is aknowledged:
o Prosecution is possible
o Vertebrates
o Anesthesia
o Use only 1 x
o Kill immediately after the experiment
o Only when necessary to advance human health

Meanwhile in Belgium/Flanders?
- Inﬂuence and input from scientist predominantly from Germany
o E. Kant
o ‘all animals exist only as a means and not for their own sakes …’
- Catholic tradition
- Strafwetboek 1867 (possession)
- No speciﬁcation (!!?) of laboratory animals or animal experimentation

,Modern Ages (1750-1950)
- Rise in animal experiments
o Infections
§ Pasteur: described rabies and they used dogs and rabbits à
mentioned to use humane eindpoints!! Ex: kill animal prior to
dying because too much su[ering
§ Koch (postulates)
o Scientiﬁcally tested drugs, vaccines,…
o Switch to using more rodent species. Rodents are pest animals, you use
catch-devides to catch them.
- Advent of psychology
o Wolfgang Köhler
o Jane Goodall
o Frans De Waal

In the UK still certain illegal experiments were done. Te procedure was condemned as
cruel and unlawful by the National Anti-Vivisection Society.
- Discussion shifted to preventing harm ... Rather than questioning the value of
animal research …
- Anesthetics
o … rise in animal use (carte blanche)
o ... Often failed to work

Russel and Burch (1959)
- “The principles of humane experimental technique.” (re-emerged in 1978)
- They further stated, “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 e[ects of any
procedure on the animal subject.”
- They introduced the 3R’s: you ask yourself 3 questions:
o Replacement? If you can replace it with a non-animal experiments
o Reduction? If it is necessary, if you can reduce it, reduce animals, reduce
su[ering and if you have done what you could
o Reﬁnement? Do I still have possibilities to further reﬁne the experiment
and make them su[er less and make them happier?

2. 3Rs concept
Replacement:
- Non-vertebrate model
- Embryonic / larval form
- In vitro test
- Computer / in silico

Reﬁnement:

, - Cage enrichment
- Use of anaesthetics, analgetics
- In vivo imaging …

Reduction:
- Sample size calculations
- Re-use
- Reporting negative data …

A fourth R: respect?
- Now recent papers, introduce fourth R. you should also include respect. How to
keep the integrity and respect?

3. Welfare and law of laboratory animals
Tom regan
- He is an animal right activist
- The case for Animal Rights (1983)
o Extension of Kant’s intrinsic value to all species
o Rights : every animal has fundamental rights
o We have to respect those rights
- Moral agents vs moral patients
o Moral agents are those who have full autonomy and are capable (=
possessing the necessary cognitive skills) in making decisions
o Moral patients are individuals who have desires, longings, known
emotions, etc.
§ Only ‘normal’ adult humans are moral agents, all others are
‘subjects of life’ and thus moral patients who have fundamental
(universal) rights.
§ All subjects of life should be considered as a goal/end not as a
means.
§ Moral actors have the duty to respect the rights of moral patients.

Peter Singer
- Animal liberation (1970)
o Speciesism!!!
§ Does not entail equality
§ We can’t consider ourselves at higher level than a dog, a monkey,…
because that is discriminating.
§ He says su[ering of mice is equal as su[ering of human beings
• Precautionary principle
- Ultilitarianism
o One should give priority to relieve the greater su[ering
- You can still do experiments but take into account the su[ering of mice, if it is for
human health.

1986: European directive

,1986: Belgian law on animal welfare
- Mentions laboratory animal
- Mentions animal experiment
- Registration
- KB’s (implementation)
2010: New European Directive reﬁnement
Implementation of new EC (KB 2014)
2014: animal welfar moved to the “gewesten” (Flandres)

“Describe the responsibilities of staff when working with research animals and recognize the
importance of having a respectful and humane attitude towards working with animals in research. “

- Responsible for providing appropriate housing, feeding, and care for the animals according
to established guidelines and protocols. Access to clean water, suitable food, and
appropriate environmental conditions such as temperature, humidity, and lighting.
- Montor the health of the animals regularly, looking out for signs of illness or distress.
- Proper, gentle handling and restraint techniques à minimize stress and discomfort for the
animals during procedures such as injections, sample collection, or behavioral
assessments.
- Continuous training and education are essential for staff members
- Recognize the importance of having a respectful and humane attitude towards working with
animals in research. Animals are sentient beings capable of experiencing pain, distress, and
emotions. Minimize any potential harm or discomfort, providing enrichment to promote
psychological well-being, and considering alternatives to animal use whenever feasible. By
maintaining a respectful and humane attitude, staff members can contribute to the ethical
conduct of research and ensure the welfare of the animals under their care.

Describe the importance of disseminating information and study results that will promote an
understanding of ethical issues, good animal welfare, and the application of the 3Rs.

- Sharing information about research methods, ethical considerations, and study results
promotes transparency and accountability within the scientific community and to the
public. It allows for scrutiny of research practices and ensures that ethical standards are
upheld.
- Disseminating information about ethical issues and the 3Rs helps educate researchers,
laboratory personnel, veterinarians, and other professionals involved in animal research. By
increasing awareness and understanding of ethical principles and alternative methods,
researchers can make informed decisions that prioritize animal welfare while advancing
scientific progress.
- Openly sharing information about animal research practices and outcomes helps build trust
with the public. When individuals understand that researchers are committed to ethical
conduct and animal welfare, they are more likely to support responsible research initiatives.
- It encourages researchers to prioritize ethical conduct in their work.
- It fosters collaboration among researchers and institutions. This collaboration can spur
innovation in research methods and technologies that further reduce the need for animal
experimentation and enhance animal welfare.
- It contributes to the development of policies and guidelines that govern animal research.

,Chapter 2: Legislation - part 1
European level
- EU directive 2010/63/EU
o The member states who signed and ratiﬁed the directive must implement
it into their legislation.
o … EU directive 1986/609/EEG
§ was implemented in the law of 1986
§ Led to a Royal Decree in 1993 (now replaced by RD 2013, which
implemented the changes because of 2010/63/EU)

National level (Belgium)
- Law animal welfare 1986
o More on the metalevel so for all kinds of animals not only lab animals
- Royal Decree 30.11.01 Verbod op sommige dierproeven
- Royal Decree 29.05.13 Protection Laboratory animals incl annexes
- Decree 17.02.2017 Protection laboratory animals (changes)
- Decree 22.02.2017 Composition Vlaamse proefdierencommissie
- Vlaamse Codex Dierenwelzijn 2023

In a nutshell:
- The protection of laboratory animals in Belgium/Flanders is basically ensured by
the Animal Welfare Law 1986 concerning the protection and welfare of animals
and the Royal Decree of 1993 replaced by the Royal Decree of 2013 concerning
the protection of laboratory animals.

1. Laboratory animal
Animal welfare law 1986: what is considered to be a laboratory animal!:

Art 3 - 15. Laboratory animal:
1. The living cephalopods used or intended for use in animal experiments,
or which are speciﬁcally kept so that their organs or tissues can be used
for scientiﬁc purposes;
2. The living non-human vertebrates used or intended for use in animal
experiments, or which are kept speciﬁcally so that their organs or tissues
may be used for scientiﬁc purposes, including their self-feeding larval
forms, as well as fetal forms of mammals from entry of the last third of
their normal development;
3. This deﬁnition also applies to animals used in animal experiments that
are at an earlier stage of development than the one in section 15.2. if
these animals are required to survive beyond that stage of
development and are in danger of su^ering pain, su^ering, distress or
lasting harm after reaching that stage as a result of the animal
experiments performed;

, - All other animals can be considered an alternative model, for example
Drosophula is an invertebrate and you should not report on it.

Art 15.2:
- As cited, live cephalopods and live non-human vertebrates should be
considered experimental animals. They also include independently feeding
larval forms and foetal forms of mammals as of the last third of their normal
development.
- This requires further clariﬁcation:
o Fishes:
§ Are considered independently feeding when both ends of the
intestines are open, and the ﬁsh takes in food
§ This time varies between species and depends, among other
things, on the temperature at which they are kept
• e.g., larvae of zebraﬁsh, kept at +28°C, reach this stage
from the ﬁfth day after fertilisation which corresponds to
120hpf. However, several factors a[ect when this stage is
reached; therefore, this cannot be seen as an irrefutable
limit.
o Amphibians:
§ Are considered independently feeding from the time when it starts
actively moving in search of food (and the yolk is largely used up)
§ This time varies signiﬁcantly between species
• e.g., clawed frogs: according to tables, these are
independently feeding from NF stage 49 onwards
o Cephalopods:
§ Are considered independently feeding immediately after hatching
o Birds and reptiles:
§ When they hatch or are born alive, whether by human action or
not, are considered experimental animals
o Mammals:
§ In the last third of their normal development (i.e., last third of
gestation) are considered experimental animals

The origin
Art. 23. § 1:
- The Flemish Government can lay down rules regarding the origin of the
experimental animals and determine special conditions with regard to keeping
laboratory animals of di[erent categories. The Flemish Government can also
prescribe rules to determine and monitor the origin of the animals. However,
dogs and cats must be registered with an indication of their origin.
- They demand that certain species that are very frequently used in experiments
have to be purpose bred.

RD2013 annex 1: purpose (scientiﬁc use)-bred (license for breeding):
Mouse (Mus musculus)
1. Rat (Rattus norvegicus)

, 2. Guinea pig (Cavia porcellus)
3. Syrian (golden) hamster (Mesocricetus auratus)
4. Chinese hamster (Cricetulus griseus)
5. Mongolian gerbil (Meriones unguiculatus)
6. Rabbit (Oryctolagus cuniculus)
7. Dog (Canis familiaris)
8. Cat (Felis catus)
9. All species of non-human primates
10. Frog ((Xenopus (laevis, tropicalis), Rana (Temporaria, pipiens))
11. Zebra ﬁsh (Danio rerio)

ð buy them by a licensed breeder, and they should be bred only to be used in an
experiment, you cannot catch a mouse and use it in your experiment.

RD2013: Purpose bred:
- There is a need for certain species of vertebrate animals used in procedures to
be bred speciﬁcally for that purpose so that their genetic, biological and
behavioral background is well- known to persons undertaking the procedures.
Such knowledge both increases the scientiﬁc quality and reliability of the
results and decreases the variability, ultimately resulting in fewer
procedures and reduced animal use.

- Exemption in case animals in annex 1 are not available in su^icient numbers
o But necessary to proof this and
o Guarantee that animals were bred in good circumstances and
o Guarantee that with these animals the scientiﬁc goals can be met and
with the same numbers of animals
o Approval from government necessary

RD2013: Non-human primates (NHP)
- Having regard to the present state of scientiﬁc knowledge, the use of NHP in
scientiﬁc procedures is still necessary in biomedical research. Due to their
genetic proximity to human beings and to their highly developed social skills, the
use of NHP in scientiﬁc procedures raises speciﬁc ethical and practical
problems in terms of meeting their behavioral, environmental and social needs
in a laboratory environment. Furthermore their use is of greatest concern to the
public. Therefore the use of NHP should be permitted only in those biomedical
areas essential for the beneﬁt of human beings and for which no other
methods (nor animal species) are yet available.

- Adopted in art 6 of KB - Biomedical areas for which NHP can be used (when
no other method is available) are deﬁned:
o Basic research
o Applied research but only for the study of disorders and diseases
o Research in the development of drugs, treatments (incl safety)
- No use of NHP for: protection of the environment, conservation of species,
education, forensic research

, RD2001:
- Non-human primates can be used in animal experiments but NO animal
experiments are allowed on Chimpansee, Bonobo, Orang-Oetan and,
Gorilla.
ð They investigate and study these species. Are these experiments? => we only talk
about experiment if it is harm or stress causing.

RD2013: What about animals not listed in the annex?
- Must not be purpose bred, but bred by a licensed breeder, supplier
o This assures some kind of quality and reduces variability.
o except farm animals (no license necessary) (not purpose-bred, no
licensed breeder)
- (Art 5) Can endangered species be used? No
o Exemption possible
§ Proof that no other animal (or alternative method) can be used
§ Only for purposes: Basic research, Applied research, Development
of drugs and Preservation of species.
- (Art 7) can animals be ‘caught’ in the wild? Yes but
o License sould be requested and approved by the government and
§ ‘proof’ that no other animals (or alternative methods) can be used
§ Caught by a competent person
- (Art 8) Can stray or ferral animals be used ? No
o Exemption possible
§ Proof that the animal is the subject of the scientiﬁc question
§ And that no other animals (or alternative methods) can be used

Animals taken from the wild:
= a previously free-living animal that has been captured or otherwise brought
under the control of man.
- Catching, handling, restraining or transporting animals should not be
considered an animal experiment (even if it exceeds the threshold, e.g. when the
animals have to be sedated to do so), unless the scientiﬁc procedures are aimed
(at least in part) at investigating the catching, handling, restraining or
transporting of animals. Nevertheless, even when this does not constitute an
animal experiment, it may involve laboratory animals for which the most reﬁned
methods must always be used with a view to the welfare of the animals
concerned.
- Many identiﬁcation methods do cause pain, but again this is not a regulated
procedure.
- Equipping the animals with a tracking device
o Using a non-invasive method is not an animal experiment, except when
it is expected that the threshold will be exceeded.
o Equipping the animals with a tracking device by an invasive method (e.g.
transmitter implantation, ...) is considered an animal experiment if the
threshold is exceeded (which will usually be the case)
- Animal experiment vs. Laboratory animal

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