At the end of this module you should:

Be able to argue the pros and cons of using animals as models for biomedical research
Be aware of the applicable legislation at the European/Spanish/Catalan context
Be familiar with the 3 Rs (replacement, reduction and refinement)
Understand that transparency about the use of animals in research is essential

1. History
2. Pros and Cons of animal research
3. Ethical issues and current debate
4. Guidelines and Regulations - the 3 Rs
5. Moving forward - towards top quality animal research

The use of animals in human research has a long history

• 2nd Century Galen of Pergamon uses animals for anatomical studies.
• 12th Century - Avenzoar introduced an experimental method of testing surgical procedures
before applying them to human patients. He performed the first experimental tracheotomy
on a goat.
• 1774 Benjamin Jesty, a farmer, inoculated pus from cowpox udder’s lesions into his sons
and wife
• 1798 Edward Jenner introduced the concept of inducing immunity to prevent diseases. The
first vaccine for smallpox was produced.
• 1849-1936 Ivan Paulov described conditioned reflex actions.
• 1885 Louis Pasteur discovered the vaccine for carbuncle.
• 1843-1910 Robert Koch discovered the bacillus responsible for tuberculosis using guinea
pigs to culture it.
• 1908 Margaret Floy Washburn published a book compiling her an other’s research on
experimental work in animal psychology, using a range of animal models
• 1909 Paul Erlich discovered chemotherapeutical agents against syphilis testing them in
rabbits.

Why animals are needed in biomedical research

• Biological similarity to humans – animals suffer from similar diseases to humans

• The short life span of some research animals allows to study them throughout their entire
life cycle
• Design of models that facilitate the study of specific diseases
• As an alternative to doing experiments with humans – although animal research also
raises some ethical issues.

Experimental animals - some numbers…

Number of animals used in 2020 in:
Catalonia: 194.878 / Spain: 675.822 / Europe: 8,6M

In the EU = Animals used for Research and Testing down nearly 22 % from 2008 to 2017. It
decreased an additional 7,48% from 2019 to 2020

The biggest group of animals used is rodents representing over 57% of all. Fish accounts for over
27% (2020 data). The last use of great apes in EU for animal research was reported in 1999.
Huge advances in medical science have been possible because of research on animals.
However the translation of the results of research done in animals to humans is not so
straightforward…

How often do highly cited animal studies translate into successful human research?
Study: the authors conducted a systematic review of animal studies specifically designed to model
human diseases. They screened articles (n=2000) published between 1980 and 2000 from 7
leading journals - Science, Nature journals, Cell etc…
76 animal trials selected, no animal study was negative, and the median citation count was
889 (639-2233).

Results: of the 76 selected studies:

28 (37%) replicated in human randomized trials (median time to replication was 7 years). 8
replicated interventions were approved for patients.
14 (18%) human studies contradicted animal studies
34 (45%) remain untested

Only about 1/3 of the highly cited animal research translated at the level of human randomized
trials

The complex extrapolation to humans of the results obtained in animal studies

• Animals are highly imperfect human models (Akhtar A. Camb Q Healthc Ethics. 2015)
• Animals can not be used as surrogate humans in drug safety testing.
• Mice seem to be a very poor model for human diseases. How many times have cancer
cures been observed in mice?
Utopian or non-realistic experimental models.
• Very young animals, without co-morbidity
• Interspecies variability
• Reductionism and lack of natural models (experiments)
Poor designs in terms of trial methodology
• Inadequate power
• Lack of randomisation and blinding procedures
• Sex bias (e.g. use of only male mice)
• Housing conditions (ie. effect of housing temperature on research results)

ANIMAL TESTING – ETHICAL ISSUES AND CURRENT DEBATE

Animal experimentation – A philosophical perspective

Is animal experimentation ethically right/wrong? This question has raised a lot of debate among
philosophers… Some influential philosophers on animal experimentation are:

AGAINST animal research:

• Peter Singer: Utilitarian argument
◦ Animal Liberation (1975) – considered by many as the bible of the modern animal rights
movement.
◦ In Defense of Animals (1985)
• Tom Regan: Deontological approach
◦ The Case for Animal Rights (1983)

FOR animal research:

• Carl Cohen:
◦ The case for the use of animals in biomedical research. NEJM 1986; 315: 865-70.

The utilitarian argument – promoted by Peter Singer

The key moral question related to animals is not whether they can reason, but whether they can
suffer (from physical pain, deprivation, loneliness, etc…). Singer echoes an assertion made by
Jeremy Bentham (1748-1831): “The question is not, can they reason? Nor can they talk? But, can
they suffer?” The principle of equality: we should give equal consideration to the suffering of
individuals, regardless of their species. Failure to do so amounts to “species-ism”, an offense
analogous to racism or sexism. Singer supports animal experimentation if the benefits obtained
outweigh the harm done, but no animal experiment should be conducted unless it would also be
permitted on a human (pain viewed with an equivalent moral cost).
The deontological approach - promoted by Tom Regan
The focus is on the RIGHTS of animals as a philosophical concept, in contrast to the utilitarian
ethics of Singer. Living sentient beings (including animals) have moral rights (eg. the right to
life) and they have these rights independently of considerations about the value of their
consequences. These rights are absolute and should never be overridden by other moral
considerations. All Individuals have equal rights, but not all animals possess the same “inherent
value”. BUT to insist that only humans have inherent value is species-ism.

Philosophers DEFENDING Animal Experimentation

Carl Cohen, dismisses both the Rights (Deontological) and the Suffering (Utilitarian)
arguments:

“Rights arise, and can be intelligibly defended, only among beings who actually do, or can, make
moral claims against one another. Whatever else rights may be, therefore, they are necessarily
human; their possessors are persons, human beings” “It does not follow from this, however, that we
are morally free to do anything we please to animals. Certainly not. To treat animals humanely,
however, is not to treat them as humans or as holders of rights”

“The elimination of horrible disease, the increase in longevity, the avoidance of great pain, the
saving of lives, and the improvement of the quality of lives (for humans and for animals) achieved
through research using animals is so incalculably great that the argument of these critics,
systematically pursued, establishes not their conclusion but its reverse: to refrain from using
animals in biomedical research is, on utilitarian grounds, morally wrong.”
Cohen, C. The case for the use of animals in biomedical research. N Eng. J Med, 1986

And… what is public opinion on this issue?

Large percentages of the general public (especially young people) express support for the
concept of animal rights, while simultaneously eating animals and supporting the use of
animals in biomedical research. Most working scientists support the use of animals in biomedical
research and are also concerned that such research be conducted humanely.

According to an IPSOS Mori survey in UK in 2016, the majority of people accept animal
research for medical purposes when there is no alternative and if there is no unnecessary
suffering to the animals. At the same time, they think there needs to be more work done into
alternatives to using animals in scientific research.

Emergence and very brief history of the animal rights movement

1875: First animal protection society was founded (in the UK), the Victoria Street Society for the
protection of Animals Liable to Vivisection, being its principal aim to stop animal research.
Vivisection became a matter of public debate (not only philosophical).
1900 – 1970’s: Antivivisection movements lose public support. Some reasons:
Advancement in medical knowledge during the 20th century changed perception of the public
toward animal testing.
Emergence rodents as models in research (instead of using domestic animals).
Socio – economic reasons (wars, recession, etc).
1975: Publication of “Animal Liberation” by P. Singer. Rebirth of the animal rights movement.
Animal rights organisations were particularly active (sometimes even violent) during the 70-90’s.
Nowadays their actions are more directed towards advocacy and influencing public policy. An
Example: Stop Vivisection initiative: Proposal to the European Commission to ban animal
experimentation in the EU in 2015. Read about the resolution of EU after the public hearing here.
Organisations defending or explaining the use of animals in biomedical research
There are also organisations defending the use of animals in research. Their aim is to provide
information about how research with animals is performed nowadays and its relevance for the
scientific and medical advancement. Check the links below to know more about some of them:
• European Animal Research Association (EARA); Understanding animal research;
AnimalResearch.Info

What effect does animal activism have on scientists?

Should individual researchers be more proactive explaining their research with animals to society?

GUIDELINES AND REGULATIONS =========================================

Legislation to manage animals in research is evolving

Much of the early debate on animal research took place in the 1980s before the introduction of
regulations and controls.
At present there are guidelines and laws that define the minimum standards that scientist must
follow in their care and use of animals.
In Europe, people working with animals must receive formal training, housing and transport of
animals is regulated and there are ethical committees and governmental institutions that revise
the protocols and control that the research is justifiable.

Three principles now govern research using animals – The 3Rs

The 3 Rs were described by Russell WMS and Burch RL in 1959 (Principles of Humane Animal
Experimentation).
The 3Rs have been internationally adopted as the principles governing animal research and have
been incorporated in international and national regulations.
The 3 Rs :

Replacement -------------------------------------------------------------------------------------------------------
-- Refers to the attempt to substitute by insentient animals, or if this is not possible, by species that
might be less susceptible to pain and distress.
Some replacement alternatives are:
Computer models
In vitro cells and tissue cultures
Organoids
Human volunteers
Replacement alternatives can be divided in:
Full replacement: avoids the use of any research animals.
Partial replacement: includes the use of some animals that, based on current scientific
thinking, are not considered capable of experiencing suffering (Invertebrates such as D.
melanogaster or C. elegans, or microorganisms).
Replacement methods for testing cosmetics: In the EU it is forbidden the use of animals for
testing cosmetic purposes since 2013 (in the UK since 2008) – alternative methods have been
developed and validated to totally phase out the use of animals in the development of cosmetic
products.
Replacement methods for household products: In the UK it is forbidden to test household
products on animals since 2015.
In the development of new drugs:
• A of new drugs fail in clinical trials even following success in animal tests (would a
reduction in animal tests change this figure?)
 • High-throughput screening in pharmaceutical industry has reduced the need for tens of
thousands of animals.
• New micro-dosing human assays can be a replacement/reduction alternative, but the
technology needs to be validated.
Until new replacement techniques are available animal research is still necessary
• Computer modeling can be good to predict protein folding, but isolated human cells will
never be able to tell us the full story about the regulation of blood pressure.
• Many in vitro studies, such as the Ames test, which assesses the mutagenic potential of a
chemical, give many false positives.
• Human tissue, cell cultures and computer modeling cannot reproduce models of mental
health problems such as depression, autism or learning disorders.
• To phase out the use of animals, more risk will need to be transferred to humans (!)

Reduction ----------------------------------------------------------------------------------------------------------
-- Attempt to use the minimum number of animal lives necessary to answer the research question.
But a design with too few animals that cannot reach statistical significance is equally wasteful and
morally troubling.

Ways of reducing the animals used in experimentation:

Right choice of animal species and strains; Exhaustive literature research to avoid repetition of
experiments; Adequate statistics and experimental design; Sharing data of studies performed in
animals

Refinement --------------------------------------------------------------------------------------------------------
-- Refers to the attempt to reduce the incidence or severity of pain and distress experienced by
laboratory animals (appropriate drugs and trained personnel).
• Anaesthesia (before and during surgery)
• Analgesia (in case of pain)
• Euthanasia
• Socialisation
• Acclimatisation after transport
• Avoid too long starvation
• Blood samples: saphenous vein instead of retroorbital punction
• Environmental improvements
◦ Enough space
◦ Social groups vs. Isolated animals
◦ Environmental enrichment

Legislation to manage animal research in Spain

• European Union: DIRECTIVE 2010/63/EU on the protection of animals used for scientific
purposes. The EU Directive is unique in the world because it sets as its ultimate goal the
full replacement of use of animals for scientific purposes.
◦ Regulates the use of animals (vertebrates and cephalopods) for scientific and educational
purposes. It regulates the origin, breeding, care, housing and killing of animals. It also
regulates the procedures to follow to experiment with animals. It specifically highlights
the 3 Rs – reuse of animals when possible, programmes for sharing tissues and organs,
and seeking of replacement methods.
• Spanish:
◦ Royal Decree 53/2013 on the protection of animals used for experimentation or other
scientific purposes – Applying the EU Directive.
◦ Orden ECC/566/2015 – establishes the skills and training requirements of the personnel
manipulating animals for experimentation and other scientific purposes.
 • Catalan: Decree 214/1997 regulates the use of animals for experimentation and other
scientific purposes, as well as the functions and the composition of “Comité Ético de
Bienestar Animal” similar to the “Institutional Animal Care and Use Committee” (IACUC)
in USA.

What does the PRBB code say about it?

8.9. Research involving experimental animals
In accordance with national and European regulations, all procedures using animals must be
previously approved by the Ethical Committee for Animal Research (CEEA-PRBB) or the
applicable body. All animal protocols must be carried out in an accredited animal facility and by
trained and accredited personnel.

The PRBB Animal Facility

4500 m2. Includes 5 independent areas, possibility of housing specific-pathogen-free animals.
The animal facility can accommodate up to 70,000 mice, 50,000 zebra fish and 500 frogs.
It includes special services for cryopreservation of mice germinal cells (embryos and sperm), and in
vivo non invasive imaging systems by optical imaging.
It has been certified by the AAALAC (Association for Assessment and Accreditation of
Laboratory Animal Care International,)

The dual role of animal facility professionals:

• Protect the animals, enforce research standards and rules and promote research.
• Manage the housing, feeding and care of animals (veterinary and nonmedical) .
• Provide training, advice and technical support for those involved in the care and/or research use
of animals.

Approval process for research involving animals at the PRBB

An animal research SUBMISSION should discuss:

• What types of procedures are being proposed and why the research is important.
• The rationale of the experiments and a justification of the species selected.
• An explanation of why other alternatives (cell cultures, computer models…) were rejected
or were inappropriate for the aims of the study.
• Steps taken to make the proposed experiments less invasive or to make use of a species
lower in the phylogenetic tree.
• A justification of the appropriateness of the number of animals requested for the
experimental series planned (with a statistical analysis if possible)
• A documented review justifying that the experiments proposed do not unnecessarily
 duplicate previous work.

Animal research PROCEDURES should describe:

• The use of anaesthetics, analgesics or sedatives when appropriate with an assessment
anticipated pain and distress.
• The justification, criteria and procedure that will be used to remove animals from the
study or to euthanise them.
• The training and experience of the personnel who will be involved
MOVING FORWARD…
**Note that the European Parliament voted in 2021 to accelerate the phase-out of animal
research.

Towards top quality animal research

Replacement, Reduction and Refinement BUT ALSO… Standardisation, prospective
registration, randomisation, systematic reviews and transparency

• Development of “Uniform Requirements” for animal research publications – Although

there are guidelines (ARRIVE, ICLAS, etc), they need to be unified and adopted by all
journals.
• Rigorous systematic reviews prior to designing new clinical trials based on animal
research.
• Enforce an animal trials registry similar to that existing for human clinical trials (to
ensure that results are reported and to avoid publication bias).
• Validation methods internationally recognized by different countries (to avoid
redundancy in the number of animals used).
• Promote a healthy debate and encourage transparency about how research with
animals is done.

Transparency on animal research

Some initiatives to increase transparency on animal research:

• Basel declaration (2011) – Aims to bring the scientific community together to advance
in the implementation of ethical principles when doing research with animals and to call
for more trust, transparency and communication on the sensitive topic of animals in
research.
• National transparency agreements: the Concordat on Openness on Animal Research
(2014) in the UK and similar transparency agreements in Spain (in 2016) and other
countries.
And some specific actions:
• The Lab Animal Tour, virtual visits to 4 animal facilities in the UK.
• At PRBB: a transparency web and training on animal research communication

More openness on how animal research is performed helps to debunk false myths… as for
example:
Alternative research methods can replace the use of all animals testing in biomedical research.
◦ Although there’s been a lot of advancement finding non-animal techniques, in most cases
there are no real alternatives, only adjunct methodologies.
Much research with animals is unnecessary.
◦ Scientists do not want to repeat old experiments. They want to make new discoveries.
Some repetition is necessary to verify and confirm results in different laboratories.
Moreover unnecessary research involving animals would not be accepted by the ethical
committees.
Most research with animals is performed on dogs, cats and monkeys.
◦ Most of the animals used in biomedical research are mice, rats and other rodents.
There are no laws or government regulations to protect research animals.
◦ There are European and national laws protecting animals used in biomedical research.
Dogs and cats (and other pets) lost or captured are sold to laboratories for research.
• Pharmaceutical and university researchers use animals bred for research from licensed
dealers.
Research animals are systematically kept in pain
• The majority of biomedical research (≥62%) does not involve the issue of pain. In another
32%, animals receive anesthetics or analgesics to minimize pain.
Researchers are indifferent to the well- being of animals or torture animals during
experimentation.
• Good science and good care are inseparable. Proper care and treatment of animals in
research are the only ways to ensure reliability and replicability of research results.
Animals are used for testing cosmetics
• Testing of cosmetics and cosmetic ingredients on animals has been banned in the UK
since 1998 and in Europe since 2013.