Severity Assessment framework EU 12 RAT Flashcards
Can prospective severity be influenced by the species being used?
Yes
A procedure is repeated multiple times, as part of a multistep procedure in the same animal. Each repetition of the procedure :
May increase or decrease in severity. when repeating procedures there is the possibility of habituation, resulting in a reduction of pain, suffering or distress, but there could also be an increase because of anticipation of the repeated events. If the repeated procedure was prospectively assessed as severe, then repeating it could increase the assessment to above the permitted upper threshold.
The assessment of the severity of a procedure is legally required to be undertaken:
before starting the procedure.
At the end of the procedure for all animals
Severity assessment of scientific procedures on laboratory mammals need to be carried out:
In all animals, including fetal forms after the start of the last third of their normal development.
An animal receives a subcutaneous injection of a compound that causes no harmful effects. It then undergoes a major surgical procedure under general anaesthesia, and the procedure is completed without the animal recovering from the anaesthetic. The prospective severity of this procedure would be:
If the procedure consisted solely of surgery under general anaesthesia without recovery, then it would be classed as “non-recovery”. In this example, a first step to the procedure was injection of a test compound, and this would be classed as mild. Overall then, the procedure is prospectively assessed as of mild severity.
what is a procedure?
A procedure is defined as:
Any use of an animal which may cause pain, suffering, distress or lasting harm.
The use must be for experimental, scientific or educational purposes. A procedure involves a combination of one or more technical acts carried out on the animal.
What is the upper threshold of severity according to to the EU
Procedures involving severe, enduring pain and distress that could not be alleviated. Such
procedures would not be permitted without exceptional justification
When should animals not be considered for re-use?
If the previous step or steps in the procedure could affect the scientific outcomes
of the new use, then the animals should be humanely killed, not reused
When may mice be re-used when the previous experiment was severe?
. 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.
• The animal has been examined by a veterinarian to ensure it has fully
recovered from the first procedure
How would you describe a non-recovery procedure?
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, provided a stable and appropriate
depth of anaesthesia is maintained. But the techniques undertaken will still cause
harm to the animal and so need to be regulated. So the category of “nonrecovery” is used
How would you describe a non-recovery procedure?
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, provided a stable and appropriate
depth of anaesthesia is maintained. But the techniques undertaken will still cause
harm to the animal and so need to be regulated. So the category of “nonrecovery” is used
What is a prospective assessment?
Prospective assessments of severity are made by the research worker before
any procedures are conducted, but they are also carried out during the evaluation
of a project by the competent authority and during the review of a project
proposal by the Animal Welfare Body. The severity category that is assigned is
based on the most severe effects likely to be experienced by an individual
animal after all refinements have been applied. It should also include the
proposed means of monitoring the animals to assess the harms, any scoresheets that will be used and interventions and end-points.
What is a retrospective assessment?
Directive 2010/63/EU requires retrospective assessment of some Projects. This
process differs from the assessment of the actual severity of all of the animals
used on each procedure, something that is required for all projects.
• The actual severity of the harms inflicted on each of the animals used must be
assessed and the actual numbers of animals and species used must be
recorded.
• An assessment is needed as to whether the objectives of the project were
achieved.
• The process should also include identification of opportunities for further
reduction, replacement and refinement of animal use in the project.
• It also involves an evaluation of the scientific outcomes of the project.
How are harms best assessed?
The effects of harms on animals are best assessed by evaluating the
animals themselves, rather than using anthropomorphic or anthropocentric
criteria. Making a judgement by considering how we would feel in similar
circumstances may often be very misleading.
What are humane endpoints?
Humane endpoints are used to terminate research on an animal at a point
where sufficient information has been gained to predict a later scientific
endpoint (such as death in toxicity testing). Humane endpoints can also be
used to provide an upper limit to the suffering experienced by the animals
when the suffering is so great that continued use of that animal can no longer
be justified.
Why should severity assessment be an ongoing process?
first time we conduct a study, we will be able to make some predictions of the
likely outcome based on similar models that we have used, or which have been
described in the scientific literature. This is our prospective assessment of
severity.
• When we conduct the study, we can observe the animals we have used
throughout the procedures, and assess and record the harms caused, these
measures comprise our on-going assessment of severity. At the end of the
study, we can make an assessment of the actual severity experienced by each
animal.
• The actual severity for each animal is used in the reporting of animal use
statistics by the Competent Authority and the European Commission.
• Once the study is completed a retrospective assessment of the project may be
required by the Competent Authority or Animal Welfare Body. This requires a
review of the actual severity of the harms for each animal, together with the
scientific outcomes we have obtained and the effects of the refinements used and
the methods of monitoring the welfare of the animals. Clearly, it is good scientific
practice to conduct a similar review for all projects.
What metrics can be used in clinical assessment of animals?
• External appearance, which could be further broken down to include
the appearance of the eyes, ears, nose, coat etc.,
• Posture, movement and spontaneous behaviour
• Response to handling
• Food and water consumption
• Bodyweight
Signs specific to a particular protocol, for example, the size of a tumour, and
its appearance (e.g. ulcerated or not) could also be included.
Key points of clinical score sheets
• The type of scoring will depend on the species, the strain and the procedure
being carried out.
• The different measures included may need to be weighted (i.e. a high score in
one specific sign would carry more weight than a high score in another sign) -
but it is usually best not to rely on one single measure to determine a humane
endpoint or other intervention.
• Ease of use is an important factor - score sheets with too many measures will
be both too time consuming to use, and likely to contain irrelevant measures.
• Score sheets should also include signs of positive welfare.
• Scoring systems need to be reviewed regularly.
• Clinical signs that are noted infrequently can be removed unless they are
important for indicating a humane endpoint has been reached.
• At the end of a procedure, assessing the correlation with clinical outcomes
(e.g. death, the onset of severe signs) and pathological findings can be used
to validate and improve scoring schemes.
• All score sheets should include a section where other observations can be
recorded. This enables any new signs that are noted frequently to be
incorporated into a revised score sheet.
• Automated technologies are becoming more widely available and can be used
to speed the on-going assessment of animals.
Key points of clinical score sheets
• The type of scoring will depend on the species, the strain and the procedure
being carried out.
• The different measures included may need to be weighted (i.e. a high score in
one specific sign would carry more weight than a high score in another sign) -
but it is usually best not to rely on one single measure to determine a humane
endpoint or other intervention.
• Ease of use is an important factor - score sheets with too many measures will
be both too time consuming to use, and likely to contain irrelevant measures.
• Score sheets should also include signs of positive welfare.
• Scoring systems need to be reviewed regularly.
• Clinical signs that are noted infrequently can be removed unless they are
important for indicating a humane endpoint has been reached.
• At the end of a procedure, assessing the correlation with clinical outcomes
(e.g. death, the onset of severe signs) and pathological findings can be used
to validate and improve scoring schemes.
• All score sheets should include a section where other observations can be
recorded. This enables any new signs that are noted frequently to be
incorporated into a revised score sheet.
• Automated technologies are becoming more widely available and can be used
to speed the on-going assessment of animals.
Why is death sometimes severe and sometimes not reported?
an animal is found dead in a research facility, this could be:
• A direct result of an experimental procedure
• Due to some unrelated cause (e.g. spontaneous disease, accidental injury)
• The EU Directive 2010/63, and national legislation in Member States, does not
require the assessment of severity and reporting of the death of an animal that is
normal and has not undergone any experimental procedure. However, if animals
have died as a consequence of failures in husbandry systems (e.g. flooding of
cages caused by a failure of an automatic watering system), then it may be a
requirement to inform the regulatory authority. Such non-procedure related
causes of death should, of course, be investigated so that appropriate measures
can be introduced to prevent their recurrence.
• If an animal has undergone or is undergoing a procedure (and this includes an
animal with a genetic modification) is found dead, it is assumed that the actual
severity of that procedure is severe, unless an informed decision can be made
that the animal did not experience severe suffering before death. Factors such as
the frequency of monitoring, use of analgesia, etc. will need to be given due
consideration, together with the cause of death (assuming this can be
determined).
• If it is unlikely that death was preceded by severe suffering, the actual severity
classification should reflect the assessment of the severity of the techniques
undertaken before death.
• Death itself is not considered an adverse effect that requires reporting to the
competent authority unless it was preceded by pain, distress or other adverse
effects. Although this is controversial, it is the view taken in the Directive and one
which has been transposed into the national legislation of Member States.
• All deaths of animals in a research facility should be carefully reviewed among
those involved (e.g. scientists, animal care staff, veterinary surgeon) as soon as
possible to ensure that all relevant information is available to determine whether
the death was procedure-related and to determine an appropriate level for
reporting purposes. Whatever the cause, measures should be
taken to avoid recurrence.
What is a prospective severity assessment?
Prospective assessment: before the actual procedure (as in when the protocol
is being assessed)
What is a retrospective severity assessment?
Retrospective assessment: when the procedure is being done or completed.
(score sheet)
What is the definition of the procedure?
The definition of a procedure which is the scope of the directive is defined in the
directive and SI is as follows: “procedure” means any use, invasive or non-invasive,
of an animal for experimental or other scientific purposes, with known or unknown
outcome, or educational purposes, which may cause the animal a level of pain,
suffering, distress or lasting harm equivalent to, or higher than, that caused by the
introduction of a needle in accordance with good veterinary practice. This
includes any course of action intended, or liable, to result in the birth or hatching of
an animal or the creation and maintenance of a genetically modified animal line in
any such condition, but excludes the killing of animals solely for the use of their
organs or tissues.’
• Must reach the needle threshold to require licencing
What level of severity should not be reached?
Subject to Regulation 88(4), a user shall not perform a procedure if it involves
severe pain, suffering or distress which is likely to be long-lasting and cannot
be ameliorated
What is a non-recovery procedure?
Procedures which are performed entirely under general
anaesthesia from which the animal shall not recover consciousness shall be
classified as ‘non-recovery’
What is a mild procedure?
: Procedures on animals as a result of which the animals are likely to
experience 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 shall be classified as ‘mild
What is a moderate procedure?
Procedures on animals as a result of which the animals are likely
to experience 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
shall be classified as ‘moderate’.
What is a severe procedure?
Procedures on animals as a result of which the animals are likely to
experience 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
Describe when animals can be reused.
a user shall not use an animal already used in a procedure or procedures in a
further procedure when a different animal, which has not previously been
used in a procedure, could be used, unless:
• the actual severity of the previous procedure or procedures was classified as
“mild” or “moderate” under Regulation 20(1),
• it is demonstrated that the animal’s general state of health and wellbeing has
been fully restored,
• the further procedure is classified as “mild”, “moderate” or “nonrecovery”
under Regulation 20(1), and
• it is in accordance with veterinary advice, taking into account the lifetime
experience of the animal
• These are further defined and expanded in ANNEX VIII
What is a grimace scale?
Developed for certain species based on changes in a number of ‘facial action units’,
such as narrowing of the eyes (orbital tightening), nose bulge, cheek bulge, ear
position or changes in the position and shape of the whiskers
Example of things that can be examined for clinical assess /score sheets
Piloerection
• Isotion Behaviour
• Lowering Of Activity
• Loss Of Food & Water Intake
• Manifestation Of Pain Vocalisation, Writhing Etc..
When does killing an animal require authorisation (other than establishment and personal authorisation)?
When using a method not listed in Annex IV of the directive
is withdrawal of food for less than 24 a regulated procedure?
No
Are the use of T mazes or radial mazes regulated?
No
Is feeding an experimental diet that meets the animals needs regulated?
no
Can a procedure have multiple steps?
yes if all steps have to be carried out in the same mouse it would be a multi step procedure rather than a collection of different procedures.
when may animals be re-used if they underwent a severe procedure?
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.
The animal has been examined by a veterinarian to ensure it has fully recovered from the first procedure.
What assessments are we required to make?
Directive 2010/63/EU requires severity assessments to be made:
As part of the process of application for authorisation of a Project (Article 37 and Annex VI).
As part of the harm-benefit assessment required for project licence authorisation (Article 38) using the classification set out in Article 15.
Prospective assessment to ensure the upper limit of severity is not exceeded (Article 15).
Assessment of actual severity to enable accurate reporting of statistics of animal use (Article 54).
As part of the decision as to whether re-use should be permitted (Article 16).
As part of the decision as to whether a retrospective assessment of a project is required (Article 39).
How is actual severity assessed?
actual severity is made based on the greatest degree of pain, distress or harm experienced by each animal, and not by the average over the course of the procedure or the severity at the end of procedure.
when are retrospective assessments required?
If the project involves the use of non-human primates.
If the project includes procedures that prospectively are classified as severe.
During the authorisation process of any project, the competent authority may specify that a retrospective review will be required.
Retrospective assessment of a project requires:
An assessment of the actual severity experienced by each animal used.
An assessment of the effectiveness of the refinement measures used.
Identification of future refinements that could be used if similar procedures were to be used on a future project.
An evaluation of the scientific outcomes of the project
The prospective severity of a procedure may change depending upon the stage of development or age of the animal that is to be used.
Correct, the severity of procedures is generally considered to be lower in immature (fetal and larval forms) than in adults. The severity is also sometimes considered lower in neonatal compared to adult animals.
When reporting the severity of procedures for collation of national (and EC) annual statistics on animal use:
The actual severity is assessed for each animal used in each procedure, and these are reported for each animal.
The severity of a procedure that is considered likely to cause moderate long-lasting pain or distress should be assessed prospectively as
Severe
Non-procedure related harms:
Are required to be included in methods to refine the harms associated with a procedure
refinements to research procedures should address both procedural and non-procedural harms. 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.
What is looping behaviour?
Repeatedly summersaulting
Applying a humane endpoint to a study means that you always
Kill the animals humanely once some predetermined measures of severity have been reached or are about to be exceeded.
Kill the animals humanely once some predetermined effects have been detected that reliably predict a scientific endpoint.
Humane endpoints should be set out in a research protocol and used in the prospective assessment of severity. They should not then be changed for subsequent studies as this would affect scientific outcomes.
False
Humane endpoints should be set in advance, when possible, and they can then be used in the prospective assessment of the severity of that procedure. They should be reviewed during and after each study is completed, to check they are appropriate. They may need to be revised because they are being applied later than needed, as this would result in avoidable pain or distress and we may also obtain poorer quality research data. In addition, if we apply the endpoint too early, we may fail to obtain valid results and these animals would be wasted. Although we will need consistency in our study design, this must be balanced with the need to optimise our endpoints for both animal welfare and scientific reasons.
The assessment of the actual severity of a procedure is made by:
the investigator, together with their research team and other advisors
When assessing the prospective severity of a procedure we should assess:
Only the harms caused by the experimental techniques used. We should also assess non-procedural harms, if these arise as a requirement of this particular study (eg single housing).
When we assess the severity of a multistep procedure this should be based on:
Assessing the combined effects of all the techniques that comprise the procedure, and the duration of the adverse effects they cause.
The initial harm-benefit assessment of a procedure requires ……………………… assessment of severity
Prospective
We plan on conducting a procedure involving surgery in mice. A prospective assessment of severity of the procedure requires information on
The anaesthetic regimen to be used
Anaesthetic monitoring and intra-operative care
Postoperative monitoring and pain assessment to be used
Humane endpoints to be applied
The method of pain relief
where can information on stocking density be found?
Annex III of Directive 2010/63/EU
Body mass can be a useful measure of the effects of some research procedures. When using this metric, we should
Compare changes in weight to the bodyweight of similar animals of the same age, for example by using growth curves provided by commercial breeders of animals.
Calculate the percentage weight loss based on the weights of normal control (untreated) animals.
Should injury always be reported?
If an injury was caused as part of a procedure for a scientific purpose, then it might be reported as moderate, but this was an accidental injury, unrelated to the study. Because the injury is not related to the procedure, this mouse would not be included in any reporting of actual severity. As the mouse hasn’t started on the study, there is no actual severity to report, as it has not undergone a procedure. However, the incident should still be investigated to try to prevent any recurrence with other mice.
Several cages of mice undergo a procedure that was prospectively assessed as mild. When assessed on day 3 of the study, the mice appeared to be experiencing the anticipated mild effects. That night, the water bottle in one cage leaks and the cage floods. The mice in this cage die overnight from hypothermia. The actual severity reported for these mice should be:
Although the mice were found dead and will almost certainly have experienced severe distress before death, the incident is an accident caused by a non-procedure related event. Provided the watering system had not been modified as a particular requirement for the study, then the actual severity would still be reported as mild. However, the incident should be recorded and investigated, to try to prevent recurrence.
