Ethical treatments of non-human animals in behavioural research & teaching

Question 1

List of the ethical treatment of animals in research

Answer 1

• legislation
• the 3Rs
• choices of species & nonanimal alternatives
• number of individuals
• procedures
• endangered species
• procurement & transport of animals
• housing & animal care
• final disposition of animals

Question 2

Legislation

Answer 2

• country-specific
• investigators acquire knowledge about local legislation (the level of animal protection and enforcement of standards differs considerably between countries and regions)
• many nations / academic institutions require experiments performed on captive animals / wildlife must first be reviewed and approved by an animal welfare, animal care & use, or ethics committee of the institution

Question 3

Legislation
Animals (Scientific Procedures) Act 1986 (ASPA)

Answer 3

Act to make new provision for protection of animals used for experimental or other scientific purposes

• UK - strictest animal research regulations in world, guided by the Animals (Scientific Procedures) Act, 1986 (ASPA)
• ASPA regulates procedures carried out on protected animals (= any living vertebrate, other than man, and any living cephalopod) fro scientific or educational purposes that may cause pain, suffering, distress or lasting harm
• regulated procedures require 3 licences (establishment, project, and person licence)
• Ethical review by an Animal Welfare and Ethical Review Body (AWERB) includes review of Non-Regulated procedures

Question 4

Legislation
Animal Welfare Act 2006

Answer 4

contains general laws relating to animal welfare. It is an offence to cause unnecessary suffering to any animal (applies to all animals on common land); contains a Duty of Care to animals (= anyone responsible for an animal must take reasonable steps to make sure the animal’s needs are met - i.e. look after an animal’s welfare & ensure that is does not suffer)

Question 5

Legislation
The Welfare of Farmed Animals Regulation 2007

Answer 5

set minimum welfare standards for farm animals generally whilst kept and reared on a farm. Some more specific provisions address laying hens, calves confined for rearing and fattening, cattle, pigs, boards, sows, piglets and rabbits

Question 6

The Three Rs

Answer 6

• first defined by Russel and Burch in their book ‘The Principles of Humane Experimental Technique’
• The NC3Rs = updated definitions in line with common scientific parlance to highlight the importance of the 3Rs to modern research practices

Replacement
Reduction
Refinement

Question 7

Replacement
Basic and updated definitions

Answer 7

Basic - avoiding or replacing the use of animals in areas where they otherwise would have been used
Updated - accelerating the development and use of predictive and robust models and tools, based on the latest science and technologies, to address important scientific questions without the use of animals

Question 8

Reduction
Basic and updated definition

Answer 8

basic - minimising the number of animals used consistent with scientific aims
updated - appropriately designed and analysed animal experiments that are robust and reproducible, and truly add to the knowledge base

Question 9

Refinement
basic and updated definition

Answer 9

basic - minimising the pain, suffering, distress or lasting harm that research animals might experience
updated - advancing research animal welfare by exploiting the latest in vivo technologies and by improving understanding of the impact of welfare on scientific outcomes

Question 10

The Three Rs - Replacement
Including full replacement and partial replacement

Answer 10

• technologies /approaches which directly replace or avoid the use of animals in experiments where they would otherwise have been used
• animal models are often costly / time-consuming and may present scientific limitations, and alternative models can address some of these concerns

Full replacement - methods that avoid use of animals for research / testing, e.g. human volunteers, tissues and cells, mathematical and computer models, and established cell lines (i.e. non-animal technologies or NATs)

Partial replacement - use of animals that, based on current scientific thinking, are not considered capable of experiencing suffering (e.g. Drosophila, nematodes, social amoebae, and immature forms of vertebrates); also includes the use of primary cells/tissues taken form animals killed solely for this purpose (i.e. not been used in a scientific procedure that causes suffering)

Question 11

The Three Rs - Replacement
Under the UK’s Animals (Scientific Procedures) Act 1986

Answer 11

• Cephalopods (octopuses, squid) = protected
• embryonic and fetal forms of mammals, birds and reptiles = protected during last third of their gestation/incubation period, fish and amphibians once they can feed independently, and cephalopods at the point they hatch (Embryonic and fetal forms = protected from an earlier stage of development if they are going to live beyond the stage described above and procedure is likely to cause them pain, suffering, distress or lasting harm after they have developed to that stage)

Question 12

The Three Rs - Reduction

Answer 12

• methods minimise number of animals used per experiment / study consistent with scientific aims
• essential for reduction that studies w/ animals are appropriately designed and analysed to ensure robust and reproducible findings
• methods allow info gathered per animal in experiment to maximise to reduce use of additional animals (e.g. imaging modalities - allow longitudinal measurements in the same animal, or microsampling of blood, allowing for repeat sampling)
• reducing number of animals used must be balanced against any additional suffering that might be caused by their repeated use
• sharing data and resources (e.g. animals, tissues and equipment) between research groups and organisation can also contribute to reduction

Question 13

The Three Rs - Refinement

Answer 13

• methods minimise pain, suffering, distress or lasting harm may be experienced by research animals, and which improve their welfare
• applies to all aspects of animal use (housing, husbandry, scientific procedures)
• Examples including ensuring animals are provided with housing that allows expression of species-specific behaviours, using appropriate anaesthesia and analgesia to minimise pain, and training animals to cooperate with procedures to minimise distress

pain + suffering alter animal’s behaviour, physiology + immunology - changes lead to variation in experimental results - impair both reliability and repeatability of studies

Question 14

PREPARE Guidelines
What does PREPARE stand for?
What are PREPARE Guidelines?

Answer 14

PREPARE = Planning Research and Experimental Procedures on Animals: Recommendations for Excellence

set of guidelines for planning experiments as part of ongoing efforts to reduce waste, promote animal alternatives (3Rs), and increase the reproducibility of research and testing (produces by a group of experts from the UK and Norway, led by Norecopa)

covers 3 broad areas which determine quality of preparation for animal studies: (1) formulation of the study, (2) dialogue between scientists and the animal facility, (3) quality control of the components in the study

Question 15

ARRIVE Guidelines
What does ARRIVE stand for?
What is ARRIVE Guidelines?

Answer 15

ARRIVE (Animal Research: Reporting of In Vivo Experiments)

• recommendations to improve reporting research involving animals - maximising quality and reliability of published research, and enabling others to better scrutinise, evaluate and reproduce it
• relevant to any study involving live animals, in any area of the biosciences; primarily aimed at researchers, reviewers and journal editors of studies involving animals; also useful for ethical review boards, funders, institutions and learned societies to help promote best practice, and ensure rigorous design and reporting of animal research

Question 16

Choice of species & nonanimal alternatives

Answer 16

• appropriate and best suited for research question(s) (requires knowledge of natural history, physiology, phylogenetic relationships, and an individual animal’s previous experience)
• if procedures or housing conditions may cause pain/discomfort/stress, use the species / strain that is least likely to experience pain or distress
• majority of invertebrates usually excluded from legislation (notable exception: cephalopods) -> does not mean they don’t experience pain/discomfort/stress, but knowledge is more limited than for vertebrates -> seek expert advice and minimise potential harm wherever possible
• live animals = generally essential in behavioural research, but nonanimal alternatives can sometimes be used (e.g. video records, animal robots, computer simulations, or webcams)

Question 17

Number of individuals

Answer 17

• use minimum number of animals necessary to test the hypotheses, w/o loss of scientific rigour (3Rs) (excess in applying criterion may lead to risk of overlooking natural phenotypic variability of animal populations as well as individual personality)
• use clear, theoretically driven hypotheses, pilot studies, good experimental design and statistical test - enable several factors to be examined simultaneously reduce number of animals used, w/o compromising research objectives
• use power analyses of pilot or previously published data to estimate lowest number of animals needed to obtain meaningful scientific data (if absent, base sample sizes on related published studies)

Question 18

What should occur if animals are exposed to painful, stressful or noxious stimuli? (procedures)

Answer 18

investigator must perform are carefully considered cost/benefit analysis (partly depends upon the goal of the research)

Question 19

What are not adequate justifications? (procedures)

Answer 19

custom and practice, economic savings, convenience, or the fact that an animal might experience the same or similar stimuli in the wild are not adequate justifications

Question 20

When can poor welfare arise? (procedures)

Answer 20

poor welfare can arise because of actions causing pain of suffering, but also lack of action or denying some opportunity or resource

Question 21

Is there much literature discussing assessment of pain, suffering and wellbeing in both vertebrates and invertebrates? (procedures)

Answer 21

considerable amount of literature

Question 22

What should be used before techniques that cause physical or psychological discomfort? (procedures)

Answer 22

Use more refined procedures before using techniques that cause physical or psychological discomfort

Consider pain- or distress-causing potential of all
aspects or stages of the procedures

Pain or suffering should be (duration / magnitude) as far as possible under the requirements of the experimental design (but studies of factors such as housing or management procedures that may induce
states of stress, anxiety and fear can be a necessary part of research aimed at improving animal welfare or scientific validity)

Question 23

What should occur to minimise preparatory stress, and immediate and residual effects? (procedures)

Answer 23

Pre- and postprocedural care to minimize preparatory stress, and immediate and residual effects

Question 24

If procedures cause more than momentary, low-intensity pain, discomfort or distress, what should be used? (procedures)

Answer 24

use appropriate anaesthesia, analgesia, tranquillization and/or adjunctive relief measures unless this would jeopardize the aims of the study; seek expert advice (e.g. vet)

Question 25

What are many forms of discomfort or suffering involve?
How can this be alleviated or prevented?
(procedures)

Answer 25

Many forms of discomfort or suffering can involve experiences other than nociception (e.g. nausea, pruritus, thermal stress, social isolation, or fear) which can be taxon and context specific; many of these can be alleviated or prevented by
medication, procedures or housing conditions that specifically target those experiences -> consult with experts

Question 26

How to prevent disruption to subjects and the wider ecosystem in fieldwork procedures?

Answer 26

minimise fear, distress or lasting harm on individual animals/populations/ecosystems

Question 27

What must occur if harm is likely to take place? (procedure - fieldwork)

Answer 27

cost/benefit analysis (cost = total impact on the ecosystem)

Question 28

What may have adverse consequences in a fieldwork procedure and how can we consider less harmful techniques?

Answer 28

Capture, handling and marking, fitting with data-loggers/transmitters, collection
of physiological data (e.g. blood or tissue samples, body temperature) or the
experimental manipulations may have adverse consequences -> consider
minimally invasive or non-invasive techniques

pilot investigations (direct negative impacts; longer-term or delayed effects)

Question 29

How to minimise suffering/discomfort if animals are removed from the population in fieldwork procedures?

Answer 29

minimize time in captivity; when returning individuals consider long-term costs
(e.g. loss of territory/mates, change in social status, loss of site-specific
information or changes in cognitive abilities); avoid possible introduction of
captive-borne zoonotics

Question 30

What should be thought out before marking and tagging procedures?

Answer 30

• Explore and fully consider the least invasive
  method of marking subjects, consider specific
  characteristics of the study species and study
  aims
• Natural, unique coloration patterns (skin, fur or
  plumage) may be suitable for identification
• Minimize pain and distress of any amount of
  handling or invasive measures

Question 31

What are included in minimally invasive procedures? (procedures - marking and tagging)

Answer 31

e.g. metallic or plastic leg or neck bands, temporary
supracutaneous tags such as glue-on markers, radiotransmitter harnesses or dyes,
reflective ‘bar codes’ -> Identify biocompatible adhesives and harnesses as well as
consider the life span of the tag/marker relative to that of the animal and the study

Question 32

What is included in medium invasive procedures? (procedures - marking and tagging)

Answer 32

e.g. tattooing, freeze
marking or branding, visible implant elastomer (VIE) tags or
subcutaneously implanted radiofrequency identification
(RFID) tags (e.g. passive integrated transponder (PIT) tags);
consider the use of antibiotics or analgesics

Question 33

What is included in invasive procedures? (procedures - marking and tagging)

Answer 33

e.g. ear, toe, feather or fin clips or
punches; -> only use when absolutely necessary, no
other forms of marking are possible or feasible, and full
consideration has been given to less invasive alternatives;
consider short-and long-term consequences and the use
of antibiotics or analgesics

Question 34

What is preferred as an encounter method? (procedures - aggression, predation and intraspecific killing)

Answer 34

• field studies of natural encounters = preferred over staged encounters

Question 35

What should occur if staged encounters are necessary? (procedures - aggression, predation and intraspecific killing)

Answer 35

models or audio/video playback (number of animals should be kept to the minimum and experiments should be as short as possible)

Question 36

What can change a species response to natural predators? (procedures - aggression, predation and intraspecific killing)

Answer 36

prey species can incorrectly habituate to frequent attacks with models, which may change their response to natural predators

Question 37

How can suffering be reduced in staged encounters? (procedures - aggression, predation and intraspecific killing)

Answer 37

by continuous observation with intervention to stop aggression at predefined levels and by providing protective barriers and escape routes for the subjects

Question 38

Endangered species

Answer 38

• all research on, or indirectly impacts, endangered or locally rare species must comply w/ relevant legislation + coordinate w/ official agencies responsible for conservation effort for species or population under study
• members of threatened species should not be placed at risk except as part of serious attempt at conservation
• observation alone - results in serious disturbance, including higher predation rates on nests of young, or their abandonment
• consider techniques / alternative species as first step
• consider further adverse consequences e.g. opening remote areas for subsequent access, or teaching techniques of anaesthesia and capture that might be misused (e.g. by poachers)

Question 39

What breeders and suppliers should be used? (procurement & transport of animals)

Answer 39

only use reputable breeders and suppliers

Question 40

What must methods for capturing wild animals ensure? (procurement & transport of animals)

Answer 40

minimise pain, distress and suffering, and must comply with any relevant legislation

Question 41

What should not be encouraged for trapping animals? (procurement & transport of animals)

Answer 41

if animals are purchased from local trappers, do not encourage trapping methods that cause suffering, or methods that involve killing many individuals to obtain a few live specimens

Question 42

What should not be created? (procurement & transport of animals)

Answer 42

do not create a local market for culling wildlife for profit

Question 43

What should not happen to endangered species/populations in the wild? (procurement & transport of animals)

Answer 43

individuals of endangered species/populations should not be taken from wild unless part of an active conservation programme

Question 44

What should individuals handling and transporting animals be? (procurement & transport of animals)

Answer 44

qualified and experienced in requirements of species (incl. provision of adequate food, water, ventilation, space and protection from waste). Potential sources of stress associated with transport should be identified and minimised

Question 45

What animals should be transported? (procurement & transport of animals)

Answer 45

only healthy animals should be transported

Question 46

What animals are not suitable for transport? (procurement & transport of animals)

Answer 46

young, dependent animals are not usually suitable for transport (transport with mother to avoid separation)

Question 47

How to reduce stress during loading / shipping? (procurement & transport of animals)

Answer 47

precondition animals to transport containers

Question 48

What should be considered when transporting animals? (procurement & transport of animals)

Answer 48

carefully consider modes of transport, schedules (e.g. reduce/eliminate layovers, unless desirable) and shipping containers; compliance with relevant transport regulations for the species (local, national and international)

Question 49

What should follow transport of animals? (procurement & transport of animals)

Answer 49

suitable arrangements for acclimatization and monitoring of animals

Question 50

Housing & Animal Care

Answer 50

• ‘Standard’ conditions or practices = not translatable across all species (consult most recent guidelines from relevant taxon-oriented professional societies)
• conditions / husbandry practices must mee, at least, minimal recommended requirements BUT larger or more enriched conditions improve animal welfare but also quality of science
• companions / appropriate social groups (unless it leads to aggression, suffering)
• frequency of cleaning / water turnover -> compromise between disease and stress pevention
• consider human-animal interactions (perception of humans may depend upon species, rearing history, and the nature of the interaction)

Question 51

Final Disposition of Animals

Answer 51

• distribution to colleagues
• release of field-trapped animals
• euthanasia

Question 52

Distribution to colleagues (Final Disposition of Animals)

Answer 52

don’t use same animals repeatedly in stressful / painful experiments; animals should never be subjected to major surgery more than once unless unavoidable and justifiable

Question 53

Release of field-trapped animals (Final Disposition of Animals)

Answer 53

may be prohibited by laws; can be critical to conservation efforts; assess whether releases into wild might be injurious or detrimental to released animal / existing populations; animals should be released only at the site where they were trapped (unless conservation efforts dictate otherwise)

Question 54

Euthanasia (Final Disposition of Animals)

Answer 54

as humanely and painlessly as possible; death should be confirmed before bodies are destroyed; advice from trained veterinarians / others with equivalent expertise; consider the use of tissues or carcasses (in line with 3Rs)