ACVS TRAINING

- Animal Ethics – describes right vs wrong way to treat animals

o ALL animal ethics reject the theory of speciesism (idea that we are superior to
other species)
o Animal rights =/= animal welfare
o Animal rights = NO acceptable use of animals
o Animal welfare = there are acceptable uses for animals, but have to account for
treating them ethically (shelter, pain-free, distress-free)
- Animal Alliance of Canada = largest animal rights group in Canada
- Cruelty Free Internation – largest animal rights group in Europe

Why is research a target?

- Excellent, false and misleading PR by animal rights groups
- Problem for the public is that research is an unknown
- Public also don’t trust the monitoring systems
o There are more rules that govern research than owning pets
o We also fail to share the positive impacts of animal research on everyday life
- Most people believe what the media says, not what actually happens
o Transparency on our behalf helps sway the public to support animal based
research

Requirements for animal research

- Must have faculty member or ACC approval
- Must have funding (can’t be funded w/o strong scientific merit)
- Approved animal use protocol (must be approved by ACC)

Regulatory Bodies
- University Council on animal care (UCAC) – primary body
- ACC – secondary body
o Everyone in the ACC have an equal voice
o ACC includes members of public (to garner public trust) and non-animal-based
researchers
o post-approval monitoring (audit)
- ACVS
- Ontario Ministry of Agriculture Food and Rural Affairs (OMAFRA) – provincial
o Animals for Research Act
o This is LAW – could end up with anyone in the lab ending up in jail/fined
o Audits are unannounced – look for noncompliance
- Canadian Council on Animal Care (CCAC) – federal
o They set the policies and guidelines
o Will not approve a study if there is no merit/benefit to human/animal health
o Also audit, but tend to announce
 Arrive w/ a panel member and spend a week around
o Have the power to pull the accreditation for all animal research @ university
The Three Rs
- Replace
o What alternatives to animals exist?
o Requires annual update – must reconsider this
- Reduce
o How can you reduce amount of animals needed WHILE achieving improved
statistical analysis
- Refine
o How can you improve animal life/welfare during the study?
o How to reduce pain/distress and overall, losses
- BONUS! Retirement
o If not required for science, UWO may send out animal for adoption

Categories of Invasiveness (CCAC)

A Experiments on MOST INVERTERBRATES or on live isolates (no animal protocol needed)
B Experiments which cause little or no discomfort or stress (e.g. intraperitoneal injection if 1x)
C Minor stress/pain (e.g. intraperitoneal injection if more than 1x)
D Moderate (e.g. surgery w/ recovery; e.g. capturing wild animals into captivity)
E Experiments which cause massive pain above its pain tolerance

Endpoints
- Endpoint = potential unacceptable condition which warrants intervention OR euthanasia
- Usually SOPs (standard operating procedures) are used
o SOP 100 = monitoring/tumour growth/rodents
o SOP 321, 322 = criteria for early euthanasia
- If SOPs aren’t used, criteria for early endpoints must be clearly defined in the animal
protocol  DEATH IS NOT AN ACCEPTABLE ENDPOINT at uwo
o Often means making monitoring/scoring sheets
o You must know what is normal/abnormal for the species, the strain, the individual
o E.g. “at a total score of 15, I will implement an endpoint”
o Activity/behavior in rodents is HELLA important
 E.g. rats like tunneling, they seek shelter when they can
o Appearance/posture
 Rodents don’t show when they’re sick (because they are prey lol)
 BUT hair coat and posture help determine
 Healthy = white, sleek fur with good posture
 Sick = puffy fur, some reddening
o Weight/Body condition
 Body condition ~ BMI
 BC1 – BC4

Euthanasia
- Method must be approved on protocol
- Must meet CCAC guidelines on acceptable and human methods for the species
- Often, overdose of an anesthetic or barbiturate (IP in rodents, IV in large animals)
Stress
- Reduce stress for animals!
- Only want your interventions to be the only significant stressor to the animal
- Garbage in = Garbage out
o Rodents are stressed about everything

Lecture 2
Adjuvants
Adjuvant = antigen delivery system
- Ovalbumin = antigen
- Alum = adjuvant
o Adjuvants NOT used in attenuated vaccines
o Adjuvants are required in vaccines where microbes are NOT producing more
antigen (multiplying)
Traditional adjuvants = aluminum salts (alum), oil-in-water emulsions
- Alum = Al(OH)3 and Mg(OH)2  non-toxic and water soluble
New Generation Adjuvants = TLR agonists, nanoparticles (virosomes)
Currently NINE adjuvants are approved in human vaccines

Antibody Generation
- Polyclonal antibodies = generated after immunization or natural infection
o Activate many B cell clones that produce Abs specific for MULTIPLE epitopes
on one antigen
o Serum/plasma contains polyclonal Abs
o Useful for Gamma globulin treatment = purified IgG Abs from pooled plasma
of 1000-10,000 donors
- Monoclonal antibodies (mAb) = have specificity for ONE antigenic epitope
o Generating hybridoma
 Fuse one B cell (from spleen) + one myeloma cell = hybridoma
 Culture one hybridoma per well; test epitope specificity of mAb
 Pool hybridoma’s with same specificity and freeze
 Hybridoma is immortal, so a renewable and reliable source of mAb