H1 – Legislation
1 The law about laboratory animals
1.1 What is an “Laboratory animal”?
= every living vertebrate animal including free living and/or reproducing larval forms
(bv. Exolotol) + excluding other foetal or embryonic forms, which are used or intended
for laboratory experiments. New legislation will include some invertebrae (like squid,
lobster = cephalopoda) and mammalian embryonic forms starting from the last third
of the pregnancy
1.2 What is an “animal experiment”?
= every use of a living vertebrate animal for experimental and other scientific
purposes. This includes everything that can inflict pain, suffering, disconformt or
permanent injury, including every treatment that has the purpose or consequence of the
birth of an animal in such a condition.
This excludes the least painful, in modern practice accepted methods to kill or mark an
animal (= “humane methods”) + non-experimental treatments in agriculture and
veterinary practice.
1.3 Important facts in the law
An animal should not be used more than once in experiments that inflict serious pain and
suffering. Pain, suffering and discomfort were not defined in previous legislation but
now included in the new European Directive which will be implemented in the new
Royal Decree (Belgium).
1.4 For what purpose van lab animals be used?
Production and control of sera, vaccines or diagnostics; toxicological and pharmacological
research; diagnose of diseases; education and answer scientific questions. So not for
cosmetic purpose.
1.5 Housing and taking care of lab animals
Housing conditions, enviroment, space to move, feed, water and care needs to meet the
animals needs + daily control of animals and environment (also during the weekends
and holidays) + regular control by the veterinarian (large animals) or experts (small
animals like rodents, rabbits).
1.6 Source and identification
No stray pets, lost or abandoned animals = only purpose bred by licensed breeding
companies (mice, rats, hamsters, dog, cats, primates,…). Large animals (pigs and sheep)
don’t not have to be purposed bred = purchase at a farm. If you need an animal that
isn’t available purpose bred (bv. specifiek honden ras), you have to apply for
exemption/vrijstelling at the government (difficult + strong motivation why you cant
choose other animals!).
Register must be kept of all animals coming and leaving the lab. Dogs, cats and primates
= marked individually and permanent.
,H1 – Legislation
1.7 Responsible use of lab animals
All experiments must be restricted to the absolute minimum, may only be preformed if
the purpose cannot be achieved by other methods, the choice of the species must be
seriously considered and preferably use animals with the lowest neurophysiologic
degree.
Animals must never be subject to serious pain, great discomfort or suffering. If this
cannot be avoided, the experiments should be performed under anaesthesia. If not
possible, analgesics or other should be used.
2 Degrees of pain and suffering
Mild -> administration of anesthesia, non-invasive imaging of animals (MRI) with
appropriate sedation of anesthesia, superficial procedures like tail and ear biopsies,…
Moderate -> surgery under general anesthesia and appropriate analgesia associated
with post-surgical pain, induction of tumors or spontaneous tumors that are expected to
cause moderate pain, chemotherapy or irradiation with sublethal dose,…
Severe -> toxicity testing where death is the end-point, chemotherapy or irradiation
with lethal dose, induction of tumors or spontaneous tumors that are expected to cause
progressive lethal disease associated with long-lasting moderate pain, testing of device
where failure may cause severe pain, distress or death (bv. Cardiac assist devices),…
3 What do you need before starting animal
experiments?
1) Governmental Laboratory licence = overview map of the rooms, kind of
experiments, list of species and overview of the staff (lab director, expert in the
protection of lab animals, project leader, biotechnicians and animal caretaker).
2) Permission of Ethical Committee = the committee does: evaluation planned
experiments, setting up ethical criteria concerning the animal experiments,
advising labs and the government + reporting the government. Composed of lab
director, project leader, biotechnician, veterinarian or expert end at least 2
independent members. The lab must aplly a suitable form with all the info.
3) Education = mendatory on all niveaus (project leader bv. 80 hours cours,
researcher and biotechnician 40, caretakers 25)
,H2 - Safety
1 Physical hazards
Large variety and possible damage = trauma, fire (inflammable gasses and liquids),
noise (dogs and pigs or machines = ear protection), UV light (damage skin and eyes ->
laminary flow) electricity (broken cables), radiation (feces of the treated animals,
urine,…), sharp objects (needles, scalpels). Also bite- and scratch wounds = Cats bv. can
give small but deep wounds -> hazard for damage of the tendons, joints and infection.
Capnocytophaga canimorsus after dog- or cat bite = first nausea, diarrhea, muscular
pain and malaise. Causes intravasal coagulation around the wound => tissue necrosis
(amputation). Patients with decreased resistance = general sepsis possible.
Streptobacilus moniliformis and Spirilium minus after rat or mouse bite =
respiratory distress, abscesses and general sepsis of organs. Can cause periods of fever
(rat-bite fever) and myocarditis or meningitis as complications -> symptoms immediately
or after several weeks (difficult to detect the source of the illness).
2 Allergy
Most important allergens = proteins in urine of rats and urine, saliva and fur
particles of guinea pig. Most exposure through air or through bites and scratches.
Allergy = first sentitisation to formation of specific IgE antibodies on mast cells and
basophils. After new contact, allergen binds on IgE with release of histamine and other
mediators -> effect immediately or 2-8 hours after or more.
Anafylaxis = allergy can be rarely deadly due to of an general allergic reaction. Mostly
after a bite + symptoms can develop very fast (from mild urticaria/netelroos when
mice/rats urinates on the arm to heavy life-theathening reactions).
Preventive measures:
- Screening programs = intracutaneous tests, antibody measurement in blood, contact
allergy test, lung function test,… -> yearly screening of people who works with lab
animals.
- Design of animal facility = adjustment of ventilation and filtration (lowers antigens in
air), cage cleaners (like laminary flows),…
- Work organisation = decreasing cage density (less proteins causing allergies), choice
of bedding material, giving them other tasks with less risks or with other species.
- Personal protective gear = vacuum to clean cages instead of swiping, lab coat and
gloves for contact on the skin and dust masks (mondkapje) for allergens in the air.
3 Chemical and protocol related hazards
Perform a risk analysis -> what are possible dangers? Like DEN = induction of liver
cancer, tamoxifen = cataract and abortion which is used to knock down genes in
transgenic animals, perfusion with formaldehyde,…
Protocol related hazards = involves chemicals of unknown risk + possible infectious
agents (used to evoke diseases in lab animals).
, H2 - Safety
Infectious agents can be categorised in 4 classes:
- Class 1 = no disease in healthy human (non-pathogenic to man, animal and plant).
Necessary facilities = lockable rooms, animals shouldn’t be able to escape and
information is indicated at the door. Windows are not allowed to open, no eating or
drinking, surfaces and floors or walls must be desinfectable, sink in the room, coat
hangers so that lab coats are not allowed out of the room.
- Class 2 = moderate risk, human/animal disease after exposure (oral intake or through
mucus) but spreading into the community is unlikely. Efficient treatment or prophylaxis
exist. Necessary facilities = same ass class 1 but biohazard sign at the door, sink is
operated by the elbow, laminair flow class II to handle the animals, autoclave and room
must be airtight.
- Class 3 = respiratory transmission which can cause serious or lethal disease in men or
animal. Spreading into the community is very likely but in most cases prophylaxis or an
efficient treatment exist. Necessary facilities = same as 2 but entrance through an
airlock, room must be disinfected with gas, everything is autoclaved before exiting the
room, collection system for urine and feces.
- Class 4 = agent with high risk of lethal disease in men or animal + high risk of
spreading into the community, no therapy. Necessary facilities = same as 3 but even
higher measures (personnel suit with segregated air supply, multiple showers.. but highly
exceptional).
4 Zoonosis
= infectious disease (subclinical to lethal) transmitted from animals to humans. Higher
risk with animals from the wild or conventional animals. These zoonotic infections vary
from subclinical to fatal.
4.1 Black death
Rats are the reservoir for the bacteria Yersinia pestis which transfer the disease
through fleas.
4.2 Leptospirosis (Weil’s disease, canicola fever)
= bacterial disease affecting humans and animals, caused by bacteria of the genus
Leptospira. High fever, severe headache, chills, muscle aches, vomiting, possible
jaundice, red eyes, abdominal pain,…
Some infected persons show no symptoms, when not treadted the patient can develop
kidney damage, menegitis, liver failure,… Death in rare cases. Contamination takes place
through contact with water, food, soil that contains urine of infected animals.
4.3 Hanta virus
Found in mice, transmitted trough secreta and excreta of respiratory tract and intestines.
Can lead to serious nephritis and complete loss of kidney function.
