Animal welfare and animal experimentation at ULiège

In Belgium, animal welfare has been a regionalized responsibility since 2014. There are therefore three ministers for animal welfare (Walloon, Flemish and Brussels regions). The 1986 law regulates the welfare and protection of experimental animals ^[1]and by the Royal Decree of May 2013. These national laws transpose the European Directive published in 2010^[2]on protecting animals used for scientific purposes.

With a few nuances, the rules are identical throughout the country despite the regionalization of powers.

National and European legislation also defines what constitutes an animal experiment.

"Animal experiment: any invasive or non-invasive use of an animal for experimental or other scientific purposes, the results of which are known or unknown, or for educational purposes, likely to cause that animal pain, suffering, distress or lasting harm equivalent to or greater than that caused by the introduction of a needle following good veterinary practice. This includes any intervention intended or likely to result in the birth or hatching of an animal or the creation and maintenance of a line of genetically modified animals under any of these conditions, but excludes the killing of animals for the sole purpose of using their organs or tissues;"

This means that, for example, simply looking at an animal (observing it), touching it or examining it in a medical context is not considered an experiment in the legal sense of the term.

[1]Law of August 14 1986 on the protection and welfare of animals: http://environnement.wallonie.be/legis/bienetreanimal/bienetre008.html

[2] Directive 2010/63/EU on the protection of animals used for scientific purposes: https://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:fr:PDF

The following information is taken from the brochure "Animal experimentation: so that's how they do it... in Belgium ", published in December 2014 by BCLAS (Belgian Council for Laboratory Animal Science) which can be consulted via this link.

Researchers pursue specific goals, notably in :

Fundamental research

• Understanding how humans and animals function, and the diseases that affect them.

Applied research

• Developing medicines and vaccines to improve human and animal health.
• Animal testing, which is compulsory by law before a drug is introduced onto the market, is designed to guarantee these products' maximum efficacy and safety. At the same time, public authorities also impose strict conditions of use to protect experimental animals.

Using experimental animals cannot be improvised.

The Walloon Region must approve any research laboratory, public or private, working with animals and scrupulously respect the legislative framework (European directive and Belgian legislation) which describes the conditions under which animal experimentation can be carried out.

An Animal Ethics Committee must authorize any animal research project within the company or university. This authorization must be obtained before the research procedures are launched. The creation and composition of all Ethics Commissions are subject to the prior approval of the Service Public de Wallonie (SPW), Service du bien-être animal.

The composition of the Ethics Commission must be multidisciplinary and include at least 7 people with recognized expertise in the fields of ethics, animal health, animal welfare, alternative methods and statistical testing. Most Ethics Commissions comprise a much larger number of members (16 at ULiège) to cover a wide range of research topics. A member of the Animal Welfare Department is a permanent guest of the Commission.

The Commissions operation is entirely under the control of the Animal Welfare Service of the SPW (formerly via the Federal Public Service - Public Health). Inspectors may inspect laboratories at any time and without warning.

Laboratory staff (animal workers, animal technicians) and researchers must undergo mandatory training specific to their function and ongoing training.

Finally, an " Animal Welfare Expert " is appointed for each laboratory after approval by the Animal Welfare Department, and is automatically a member of the Ethics Commission. At ULiège, three veterinary surgeons are responsible for the day-to-day supervision of laboratory animal welfare.

All animals are monitored seven days a week, all year round.

The latest technology is used for anaesthesia (gas anaesthesia, as in humans).

Various costly methods are used for rodent imaging (ultrasound, X-ray, CT scan, pet scan, nuclear magnetic resonance).

Surgery, if required, is performed in the same way as in humans, using strict aseptic methods and sterile equipment.

Medication used to control any pain is the same as for human beings (anti-inflammatories, morphine derivatives).

Regarding housing rodents, preference is given to regulated, ventilated cages. A cage for four mice costs 500 euros, for example (coupled with a sterile hood to prevent animal infections).

Animal health is also monitored by regular checks for any germs that may be present, in line with scientific recommendations in this field.

As part of a European Citizens' Initiative, 1.17 million European citizens submitted a request to the European Commission in March 2015 to repeal animal experimentation as defined by Directive 2010/63/EU.

"We urge the European Commission to repeal Directive 2010/63/EU on the protection of animals used for scientific purposes and instead present a new proposal for a directive to end animal experimentation and make it compulsory for biomedical and toxicological research to use data relevant to the human species."

Extract from the European Citizens' Initiative "Stop Vivisection", whose file is on their website.

In its response made public on June 3, 2015, the European Commission explained why it did not, at this stage, intend to repeal this directive and suggest the adoption of a new legislative framework.

" The Commission stresses that, for the time being, animal experimentation remains essential to protect the health of citizens and animals, and to preserve the environment. While contributing to the ultimate goal of total replacement of animal testing, Directive 2010/63/EU is an indispensable instrument at EU level to protect animals that still need to be used. "

" While fully recognizing the need to advance scientific knowledge before alternative formulas can be developed in all sectors where animal testing is still used, the Commission will continue to promote the development and implementation of alternative methods, encourage cooperation and knowledge sharing in all sectors, and validate new methods while facilitating their regulatory approval. In addition, the Commission will actively monitor compliance with Directive 2010/63/EU, particularly the "three Rs" principle. The Commission will remain in close consultation with the scientific community at EU and international level to find alternative testing methods. It will organize a conference by the end of 2016 on how to progress towards the goal of phasing out animal testing. "

The full response from the European Commission can be downloaded here and here.

It is important to note that animals brought to and treated at the University Veterinary Clinic are not included in animal experimentation protocols. However, important scientific data may be obtained, for example, from blood samples (or radiological images) obtained as part of the diagnostic work to be carried out after the clinical examination. If it is in the interest of animal health to use them for a scientific publication (helpful in informing other researchers), then, as in human medicine, informed consent is signed by the owners, who have therefore been notified a priori.

If, exceptionally, dogs or cats from private owners are required for the study of a specific pathology (to treat it), for the reason that they alone present this pathology (because of their breed, for example), the owners concerned are also informed, and both the Ethics Commission and the Animal Welfare Department must give them a priori agreement. The dog or cat then goes home.

ULiège is a comprehensive university, with 11 faculties and over 3200 teaching and research staff active in almost every field of knowledge. To pursue their research and generate new knowledge from which humans and/or animals will be the beneficiaries, the use of laboratory animals in experiments may be not only necessary but also compulsory by law.

ULiège is authorized by the public authorities to conduct animal studies.

Experimentation using laboratory animals is governed in Belgium by European and national legislation, which ULiège and all its approved laboratories comply with strictly and under the permanent control of the Animal Welfare Service of the SPW.zAny research project requiring experimentation using laboratory animals must receive prior approval of the ULiège Ethics Commission. Its structure and missions are defined in the 1986 law, recently amended. A member of the Animal Welfare Department is a permanent guest of the Commission.

At the same time, in the interests of transparency and public information on the scientific work carried out, the "non-technical summaries" of each research project are posted on a European Commission website. At the end of projects, retrospective analyses are also published.

Alternative methods are implemented mainly by the same researchers who develop them. In Belgium, as in Liège, we have already seen a significant reduction in the number of animals used in laboratories in recent years, in favour of computerized methods, simulations or cell cultures.

However these alternative methods cannot yet wholly replace experiments using laboratory animals. It remains a necessity for the advancement of human and animal health, as the European Commission recently reminded following the 2015 European Citizens' Initiative "Stop Vivisection!" (see "Advanced methodology").

Between 2014 and 2021, the number of animals used in Wallonia has been almost halved (286,252 animals used in 2014 versus 148,438 in 2021). The number of animals used in the Walloon region and in Belgium is published annually by the SPW.

The vast majority of experimental animals (over 87%) are rodents (mice, rats, guinea pigs) and lagomorphs (rabbits).

There is a trend towards increasing the use of fish (and especially fish larvae), in particular zebrafish (Danio rerio), which is helping to refine the species enrolled and thus replace rodents with so-called less sensitive species, as recommended by the Law. In this way, 11.2% of the animals registered are fish.

To this end, ULiège has developed a large platform of zebrafish, which are gradually replacing mice in studies of body development, cancerous and/or infectious processes and their treatment.

In the case of laboratory dogs, the main aim is to improve the health of their fellow creatures. This research, always mindful of the animal's well-being, aims to improve human health and that of our pets. One example is the experiments carried out (in France) using dogs, which in 2017 led to the publication of a new gene therapy to restore muscle strength in Duchenne muscular dystrophy in dogs[2], paving the way for the development of human treatments. At Liège, laboratory dogs are used, for example, in

protocols to study urinary tract infections, the efficacy of treatments administered through the nose to combat colds, incontinence in older dogs, prostate hypertrophy, sperm vitality, new dressings, etc.

ULiège stopped using laboratory cats for research some ten years ago. Their help has led, for example, to improvements in the treatment of asthma in this species, as well as in the treatment of ringworm, which often affects cats and also contaminates humans. All these cats have since found adoptive families within a strictly regulated framework.

There have been no monkeys at the University of Liège for some thirty years, and none in the Walloon Region for several years.

[2]https://www.nature.com/articles/ncomms16105

The numbers of animals given in the statistics do not represent the number of animals euthanized. This is an extremely important point.

For example, if 5 dogs undergo a blood test and are left alive afterwards, they will still appear in the statistics on the use of dogs in research.

So don't confuse the number of animals enrolled with the number of animals euthanized.

The figures for animals (especially dogs and cats, but also horses) enrolled (without actually being euthanized) are regularly quoted to rouse the authorities, without discriminating between the two. It's important to be precise and to provide complete information.

Vivisection" is not practised in laboratories.

Some opponents of laboratory animal experimentation often use this old term to alarm public opinion. The term "vivisection" refers to the "sectioning" of "live, unanesthetized animals". It implies animal suffering. Such practices have long since ceased in laboratories, and any manipulation that might induce pain is carried out on animals anaesthetized using the most modern methods, or on which specific analgesia has been applied. Therefore, the term "experimental surgical protocols carried out under strictly controlled general (or local) anaesthesia" is more appropriate.

The law specifies that the following species must come from an approved breeder, a guarantee of the high quality and health of these animals.

1. Mouse (Mus musculus)
2. Rat (Rattus norvegicus)
3. Guinea pig (Cavia porcellus)
4. Syrian (golden) hamster (Mesocricetus auratus)
5. Chinese hamster (Cricetulus griseus)
6. Mongolian gerbil (Meriones unguiculatus)
7. Rabbit (Oryctolagus cuniculus)
8. Dog (Canis familiaris)
9. Cat (Felis catus)
10. All non-human primate species
11. Frogs (Xenopus laevis, tropicalis), Rana (temporaria, pipiens)
12. Zebra fish (Danio rerio)

Other species, such as farm animals, may come from an approved farm.

Unless there is an exceptional dispensation from the SPW (e.g. an endangered species that needs to be saved), experimental animals may not come from the wild, where they would be captured.

Unless there is an exceptional dispensation from the SPW (for example, an endangered species that must be saved), experimental animals may not come from the S.P.A. or shelters.

The University of Liège (Faculty of Veterinary Medicine) has an approved laboratory for using Beagles.

No dogs were used in this laboratory in 2021.

Except for protocols where the animal must be euthanized before undergoing organ removal for research purposes (a common practice with mice and rats), or when the animals have not regained total health, there is no legal obligation to euthanize the animal at the end of an experimental protocol.

The University of Liège is also habitually re-homing animals when possible. Dogs, cats, horses, geckos, guinea pigs and turtles have already been given up to private individuals (subject to a rigorous adoption contract and the approval of the Ethics Commission).

These animals can only be given up if they are in perfect condition, and the Animal Welfare Expert, a veterinary surgeon, must attest to this.

The national legislation requires the various categories of personnel working with laboratory animals to undergo appropriate training, comprising a number of hours and well-defined topics.

The University of Liège organizes this training in the form of a University Certificate (a real diploma) and requires its researchers to follow it.

The same applies to the animal workers and technicians concerned.

The University of Liège promotes the Basel Declaration.

Like the Declaration of Helsinki, which permanently changed the ethical landscape of human clinical research, the Basel Declaration aims to bring together the scientific community to pursue and implement ethical principles such as the 3R's (Replacement, Reduction, Refinement) wherever animals are used.

It is also committed to greater trust, transparency and communication of activities concerning animal experimentation.

In return, the scientists who have signed it expect the community to recognize the fundamental usefulness of these experiments for medicine, now and in the future.

The University of Liège supports the Basel Declaration.

It recognizes that :

1. It is impossible to separate "basic" and "applied" research in biomedical research: it is a continuous process, from the study of fundamental physiological functions to the understanding of the principles of disease and the development of therapies
2. Animal research must continue to be authorized where necessary.
3. That new laws and regulations are only helpful if the arguments for and against them result from a democratic debate based on facts.

The University of Liège therefore, agrees that "society and legislators should condemn the actions of radical groups who use illegal means or violence (including psychological violence) against the scientific community under the pretext of protecting animals, and that members of organizations representing animal interests should be invited to discuss all critical issues openly with the scientific community.

Alternative methods" refer to any initiative and/or approach developed to help implement the 3 Rs principle (Replacement, Reduction, Refinement), a code suggested in 1959 by Russell W. and Burch L.

So it's not just about strict "replacement", by which we mean "replacing animals with other tests^[1] ".

• Replacement is the use of methods that replace animal testing with other non-animal forms (in vitro: cell cultures in culture dishes; computer and simulation models; human experiments)
• Reduction consists in reducing the number of animals used in studies (is it possible to experiment with fewer animals?For example, new imaging methods make it possible to reduce the number of in vivo experiments. Rather than euthanising the animal to study its pathology at staggered times, we can take repeated images while leaving the animal alive).
• Refinement is defined as any improvement in the methodology for using animals (is the experimental protocol organized in such a way as to minimize animal discomfort?)

European Directive 2010/63/EU states that the ultimate goal is the progressive elimination of animal experimentation, but acknowledges that the use of animals is still necessary for the time being.

This directive firmly anchors the 3R's principle in European legislation and subsequent national legislation. No animal experiment may be carried out if an alternative method not using live animals is scientifically satisfactory for achieving the desired aim.

Ethics Commissions ensure compliance with these principles.

Alternative methods and, more generally, respect for the 3R's principle have enabled us to reduce the number of animals used in experiments. At the same time, advances in research have made it possible to answer more scientific questions with the same number of animals.

[1] Rogiers V and Beken S, entitled "Alternative methods to animal experimentation", 2000

Some extracts from a carte blanche by Prof. Marc Vandenheede, published in "Le Vif" on 23/04/2018

"Using animals for scientific research often requires involving them in procedures that generate suffering and end in death. This reality has led the European Union to frame this activity with highly restrictive animal welfare legislation. (...)

However, the assurance of "comfortable" living and dying conditions for experimental animals does not entirely sidestep the fundamental ethical question of the justification for their use. This can be summed up in the following dilemma: how can we argue for the scientific validity of the use of animal models based on their proximity to humans, while at the same time using the opposite argument to defend their ethical validity?

(...)

A first step out of this dilemma is to recognize the total bankruptcy of the Cartesian animal-machine model, and thus to fully accept our societal responsibility for the use of another sentient being for our benefit (including its killing), to the point of including an ontological or even spiritual dimension.

(...)

Something concrete? Why not start by thanking experimental animals (...)? In Japan, most research centers and universities that use animal experiments have set up a monument in tribute to the animals, and regularly thank them in ceremonies involving all staff. In the U.S.A., the famous N.I.H. ("National Institute of Health") research center has erected a commemorative plaque reading: "With recognition and gratitude to the research animals and the NIH Animal Care and Use Community that have contributed to our exceptional biomedical research advances". This is also the case in Canada, at the University of Guelph for the past 30 years: "In recognition of the animals used by the University of Guelph community to support excellence in teaching and research". Other avenues can be explored, such as setting up a foundation dedicated to research alternatives for the direct benefit of animals. But why not start by simply not forgetting them during "Télévie", "Cap 48" or the next "Téléthon"?