Ethics Welfare and 3 Rs on the RAT Platform level 1

Question 1

What are the ethical principles underlying EU Directive 2010/63?

Answer 1

Animals have moral value, and so any harm to them should be avoided.

Question 2

What approach with regards to the ethical dilemma do these laws take.

Answer 2

Utilitarian

Question 3

Where can you find the list of appropriate measures for killing animals?

Answer 3

Annex IV

Question 4

How do we decide what animals to use in studies?

Answer 4

We typically decide based on a combination of degrees of sentience (non-human primates are considered of greatest concern) and public opinion (dogs and cats are popular pets).

Question 5

What type of projects doe the EU directive require authorisation before.

Answer 5

The Directive requires the authorisation of projects involving animal species that are capable of experiencing pain or distress. This includes all vertebrates, including foetal and larval forms, and cephalopods (octopus and cuttlefish). Greater consideration is generally given to those species with higher cognitive capacities (e.g. non-human primates), and the use of Great Apes, such as Chimpanzees, is either not permitted or requires exceptional authorisation at the level of the European Commission.

Question 6

What species requires exceptional authorisation?

Answer 6

Great Apes & Chimpanzees

Question 7

What is the recommendation when there is a choice of species?

Answer 7

When there is a choice of species, Directive 2010/63/EU recommends that the “least sentient” species be selected. This is based on the assumption that species of “lower” sentience have less capacity for experiencing pain and distress.

Question 8

What are the 3 R’s?

Answer 8

Replacement, Reduction and Refinement.

Question 9

Who penned the 3 R’s?

Answer 9

Russel & Burch

Question 10

What order should they be considered?

Answer 10

Replacement and reduction followed by Refinement

Question 11

Where were the 3 R’s first described?

Answer 11

the Principles of Humane Experimental Technique

Question 12

Who was the Principles of Humane Experimental Technique commissioned by?

Answer 12

The Universities Federation for Animal Welfare.

Question 13

Who Reviewed the 3 R’s?

Answer 13

Tannenberg and Bennet
Graham and Prescott

Question 14

What is Replacement?

Answer 14

Methods that avoid or replace the use of animals in areas where they would have otherwise been used.

Question 15

What is Reduction?

Answer 15

Methods that minimize the number of animals used per experiment or test, either by enabling researchers to obtain comparable levels of information (of a given amount and precision) from fewer animals, or to obtain more information from the same number of animals (thereby avoiding further animal use).

Question 16

What is Refinement?

Answer 16

Methods that minimize any pain, suffering, distress, or lasting harm that may be experienced by the animals, and improve animal welfare. Refinement applies to all aspects of animal use, from the housing and husbandry of the animals used, to the scientific procedures performed upon them.

Question 17

What is Complete Replacement?

Answer 17

Complete replacement is the use of model systems that make no use of animals: computer simulations, studies using human tissues, or immortalised cell lines.

Question 18

What is Partial Replacement?

Answer 18

Partial replacement is the use of animals or life stages of animals that currently are not considered to have a sufficient degree of sentience to be capable of experiencing pain or distress. Examples include Drosophila (shown below), nematodes, and embryonic and larval forms of species that are only protected under Directive 2010/63/EU when at a later stage of development, such as newly-hatched zebrafish larvae.
artial replacements also include using tissues from animals killed for some other purpose. For example, farm animals killed for human consumption. It also encompasses animals which were killed solely to provide tissue for studies in-vitro which had not undergone any prior scientific procedure.

Question 19

Where can information on alternatives to animals be found?

Answer 19

In the “Alternatives” e-learning modules (EU52 and EU60) hosted on the ETPLAS website(opens in a new tab).

Question 20

What are the 4 main points for reduction?

Answer 20

Good experimental Design. Optimum designs that estimate group sizes carefully will avoid the unnecessary use of animals - the number used should be neither too large nor too small.
Improving experimental design. PREPARE and ARRIVE guidelines and other tools such as the experimental design assistant.
Reduction by obtaining more Data. Reduction by increasing the data obtained from each animal. Improved methodologies offer considerable opportunities for obtaining serial measures from individual animals rather than requiring a separate group of animals at each time point of a study.
Reduction - Sharing the data or other materials obtained with other research groups. Sharing the data obtained from your work with other groups is likely to increase the benefits of your work and could produce an overall reduction of animal use within a particular area of research.

Question 21

Where can refinements be made easiest and with the most benefits?

Answer 21

By refining procedure and animal care and husbandry.

Question 22

What is a culture of care?

Answer 22

Developing policies related to animal care and use that go beyond observing regulatory requirements and implementing the 3Rs. More info can be found on the NORECOPA website.

Question 23

What are search hedges?

Answer 23

Search hedges are pre-made, standardized search strings that can be used to identify relevant articles for a specific research topic. Search hedges can help save time and make searches more effective, but they may not be available for your area of research. However, you may be able to use some as a starting point (e.g. for particular animal species, to include all the synonyms(opens in a new tab),

Question 24

What are other sources of info on the 3 R’s?

Answer 24

*AGRICOLA

• PubMed
• Web of Science
• Scopus
• BIOSIS
• Zoological Record
• EMBASE
  EURL ECVAM
  NORECOPA
  CAAT - Centre for Alternatives to Animal Testing.
  NC3R’s website

Question 25

At what levels are harm benefit analyses carried out?

Answer 25

This assessment may be made at a local, regional or national level, depending upon your national implementation of the EU Directive.

Question 26

Are potential transport harms considered in the harm benefit analysis?

Answer 26

No, these potential harms are not included in the assessment required under the Directive, but methods of reducing their impact can benefit both animal welfare and the research outcomes of the study.

Question 27

Is single housing of animals considered a harm?

Answer 27

Social isolation is stressful, although rats will habituate to this if sufficient time is allocated. Since single housing is apparently necessary to measure heart activity, it may not be avoidable for this study and should be considered a "harm". It might be possible to avoid this by redesigning the study or using alternative telemetry systems. Although it appears each rat will be housed in standard cages, it needs to be clarified if any environmental enrichment will be provided.

Question 28

Important considerations when assessing the harms of surgery.

Answer 28

Is the method of anaesthesia appropriate to the species and procedure, and is the most "refined". Is the investigator who will carry out the surgery thoroughly familiar with the technique? Are appropriate supportive measures in place, for example, ensuring the rats do not become hypothermic during the procedure or during recovery? Is an appropriate analgesic regimen being used? Is its efficacy being assessed?

Question 29

Assessing the harms associated with administration of the test substance?

Answer 29

Is gavage the most suitable, can alternative methods be used. Is the person giving the gavage suitably trained. Can the substance be given in a palatable base. Have adverse effects of the substance been considered.

Question 30

Assessing harms associated with Euthanasia?

Answer 30

Will it interfere with results to be collected? Will it be carried out competently?

Question 31

What are non-procedural harms?

Answer 31

Harms that arise not as a direct consequence of the procedure but simply because the animals are housed in a laboratory environment. These non-procedural harms do not form part of the severity assessment of a procedure unless they are required as part of the study design. In this example, single-housing is required as part of the procedure, so the stress caused by this should be included in the assessment. The Directive 2010/63/EU requires refinement of all of these harms.

Question 32

How long does it typically take rats to recover from transport?

Answer 32

7-14 days

Question 33

What are some refinements that can be made to reduce the harms of transport?

Answer 33

By reducing journey times, avoiding prolonged housing in the transport boxes before or after transport, and providing appropriate bedding, food and fluids.

Question 34

How can the harms of individual housing be reduced?

Answer 34

Animals should be allowed time to adapt to this and the changes in bedding, diet and cage environment before the additional stress of experimental techniques. Environmental enrichment should also be used, as this will reduce the stress associated with housing in a barren environment. The rats in the image above have tubes as shelters and chew sticks to promote natural gnawing behaviour.

Question 35

How can drug administration be refined?

Answer 35

The rats will receive the substance by gavage, and using positive reinforcement training can have major benefits in reducing the distress associated with this procedure. Alternatively, some materials may be consumed voluntarily in palatable mixtures (e.g. flavoured syrups). Rats can also be trained to drink voluntarily from a syringe, which is effective and results in accurate dosing

Question 36

What are the 5 freedoms?

Answer 36

Freedom from hunger or thirst Freedom from discomfort Freedom from pain, injury or disease Freedom to express (most) normal behaviour Freedom from fear and distress

Question 37

How do we assess severity?

Answer 37

We must combine the different harms in some way. The approach taken by Directive EU 2010/63 is to grade the overall severity of harms as non-recovery, mild, moderate or severe. This assessment of "severity" is then considered alongside the project's potential benefits in the harm-benefit assessment.

Question 38

Why do we assess Non-procedure related harms?

Answer 38

They are required to be included in methods to refine the harms associated with a procedure. Although non-procedure related harms are not included in the severity assessment of a project, they can cause detrimental welfare affects and impact on scientific outcomes. You should also consider the potential cumulative effects of "below threshold" techniques.

Question 39

What is a prospective assessment?

Answer 39

A prospective assessment is carried out before a project is authorised, initially by the applicant, and this assessment should be confirmed or modified during evaluation. Prospective assessment of severity requires a careful evaluation of all of the harms that might occur when the animals are used in the study. Assessments will become more consistent as knowledge is gained of the effects of different manipulations on the animal species that will be used. It is often helpful to discuss the assessment with colleagues who have experience of the model or techniques that will be used. The assigned severity category is based on the most severe effects likely to be experienced by an individual animal after all refinements have been applied.

Question 40

How is the assigned severity category decided?

Answer 40

Based on the most severe effects likely to be experienced by an individual animal after all refinements have been applied.

Question 41

How can severity be considered?

Answer 41

Severity can be considered as the number of steps on a ladder. Refinements reduce the harm the animal experiences and could move an animal down from a higher category to a lower one. Extending harms by extending the length of a procedure might move them up a level. Carrying out a technique multiple times might also move the animal up a level

Question 42

How is non-recovery described?

Answer 42

Non-recovery is used to classify procedures conducted entirely under general anaesthesia. The animal is euthanased while still anaesthetised. General anaesthesia prevents the animal from being aware of any pain, suffering or distress since it is unconscious throughout. But the techniques undertaken will still cause harm to the animal and so need to be regulated. So the category of "non-recovery" is used.

Question 43

How is mild described?

Answer 43

Mild procedures are defined as those likely to cause short-term mild pain, suffering or distress, as well as procedures with no significant impairment of the well-being or general condition of the animals.

Question 44

How is moderate described?

Answer 44

Moderate procedures are defined as those which are likely to cause short-term moderate pain, suffering or distress, or long-lasting mild pain, suffering or distress as well as procedures that are likely to cause moderate impairment of the well-being or general condition of the animals.

Question 45

How is severe described?

Answer 45

Severe procedures are defined as those which are likely to cause severe pain, suffering or distress, or long-lasting moderate pain, suffering or distress, as well as procedures, that are likely to cause severe impairment of the well-being or general condition of the animals.

Question 46

When is actual severity to be reported?

Answer 46

At the end of the procedure, these ongoing assessments enable the actual severity of each animal to be determined and reported by the research group.

Question 47

How would the same mild technique applied several times be assigned?

Answer 47

Depends on if they fully recover between steps or not. If they recover it may be assigned mild, if not depending on how bad it is, it may be assigned severe or moderate. A multi-step procedure that requires a mild procedure to be carried out several times is more likely to be classified as moderate: If it is predicted that the animal will fail to recover between each step of the procedure.

Question 48

Why is a harm-benefit analysis difficult?

Answer 48

Harms to animals cannot be quantified accurately. The benefits of research projects are uncertain. Balancing harms to one group against benefits to another makes successful analysis problematic.

Question 49

What is re-use?

Answer 49

Defining "re-use" of animals is a little complex. Suppose a project involves a series of experimental manipulations on an animal, which all form part of one multi-step procedure, all conducted for a single scientific purpose. In that case, this is not classed as reuse.

Question 50

What is a multi-step procedure?

Answer 50

This procedure consists of several linked techniques. The outcome of this multi-step procedure requires that all of the steps are undertaken in the same animal and is categorised as continuous use of the animal. We would consider all of these steps, assess the harms and include this in our harm-benefit analysis. The project might have been designed to assess the effects of several compounds in the same animals since this could reduce variation caused by using different groups of animals for each compound. This would still be continuous use. In this case, however, the study has been completed, the implants are still functioning, and if the drug is known to be excreted rapidly, these animals might be suitable for another study. But we need to consider the ethical, scientific and legal constraints associated with what is now classed as "re-use".

Question 51

When may re-use be permissible?

Answer 51

Reuse may be permissible if they are assessed as mild or moderate. If the procedure was severe, then re-use might be permitted with special approval from the regulatory authority, providing: The animal has only been used once on a severe procedure. A veterinarian has examined the animal to ensure it has fully recovered from the first procedure.

Question 52

What is required for assessment of re-use?

Answer 52

Assessment of the health of the animal by the facility veterinarian. Assessment of the cumulative effects of the procedures that have been undertaken. Assessment of the potential interaction of the completed procedure with any subsequent use of the animal. A prospective assessment that the severity of the further procedure is mild, moderate or non-recovery. Re-use should always be considered on a case-by-case basis.

Question 53

For what severity procedures can mice might not be used?

Answer 53

Severe unless special approval from the regulatory authority has been granted.

Question 54

Can the prospective severity classification of a procedure be influenced by the choice of species that will be used?

Answer 54

Yes, the prospective severity may change depending on the choice of species.

Question 55

How would a mild procedure that causes prolonged mild pain and distress be classified?

Answer 55

As moderate.

Question 56

When carrying out a prospective severity procedure on a multi-step procedure do the harms add up.

Answer 56

The prospective assessment of severity of multi-step procedures must take account of all of the harms, but these may not simply "add-up". An initial prospective assessment will need to be made, but it may not be possible to assess severity accurately until the actual effects of the steps on the animal are assessed during the procedure.