Animal experiments

Animal Ethics Checklist

The Editorial Boards endorse the ARRIVE Guidelines for reporting in vivo experiments:
http://onlinelibrary.wiley.com/doi/10.1113/jphysiol.2010.192278/full
http://onlinelibrary.wiley.com/doi/10.1113/expphysiol.2010.053793/full

Authors of papers submitted to The Journal of Physiology and Experimental Physiology must adhere to the journals’ principles and standards for reporting animal experiments.
Authors wishing to submit to either of the journals should read these guidelines to ensure that their research and experiments adhere to our exacting standards.
Authors are required to provide the reference number from their Institutional Ethics Committee approval form on submission. If no number has been given a dated 'Letter of approval' must be provided. Experimental Physiology authors must include this information at the beginning of the Methods section of their manuscript.A copy of the full form detailing experimental protocol might be requested for review purposes.

Euthanasia/humane killing: Animals must be killed using methods approved for that species, stage of development and size. The standards required by the journals are those of the UK/European Union legislation, set out in in ASPA Schedule 1 in the UK and in Annex IV in the European Directive 2010/63/EU. Methods not on Schedule 1 or Annex IV could be permissible but even where the method used conforms with national guidelines, the onus is on the author to provide scientific evidence to demonstrate that it is at least as humane as those on Schedule 1 and Annex IV.

Our guidelines have been outlined in the following editorial ‘Principles and standards for reporting animal experiments in The Journal of Physiology and Experimental Physiology'.

Confirmation of compliance
Authors must include in the Methods section a statement that the investigators understand the ethical principles under which the journal operates and that their work complies with the animal ethics checklist as outlined in the editorial and copied below.

Ethical approval

At the beginning of the Methods section state the institutional ethical committee that approved the study and also the national guidelines under which the institution operates. Authors must demonstrate they have taken all steps to minimise the animals’ pain and suffering.

Ethical information must be included for each manuscript. It is not sufficient to refer to previous publications for details.

Origin and source of the animals or tissues

Identify the animal supplier/commercial breeder. For farm animals it is not necessary to name the commercial supplier. However information on the country's regulatory authority governing animal husbandry and transportation must be provided. For example: “All animals were reared and transported under conditions specified in the UK’s Animal Welfare Act 2006 and The Welfare of Farm Animals (England) Regulations 2007.”
Specify species, strain, genetic background, weight, sex and age. Indicate the group size and total numbers of animals, including any animals used but subsequently excluded for any reason, specifying any unexpected events.
In studies of isolated tissues, including cell cultures, details of the methods by which these tissues were obtained, including the method of anaesthesia or killing. Always include the experimental temperature for in vitro studies.

Access to food and water

State the feeding regime e.g. fasting, feeding ad libitum or on a specific diet.

Euthanasia

Animals must be killed using methods approved for that species, stage of development and size. If national or local guidance on this is not available, information on UK/European Union requirements can be found in ASPA Schedule 1 in the UK and in Annex IV in the European Directive 2010/63/EU. Methods not on Schedule 1 or Annex IV could be permissible but the onus is on the author to provide scientific evidence to support the method used and to demonstrate that it is at least as humane as those on Schedule 1 and Annex IV.

Anaesthetic protocols & monitoring

Describe anaesthetic protocols in detail (premedication, anaesthetic(s) used, dose, route, supplementary dosage). These must be appropriate for the species, experimental time course and whether the procedure was terminal (in which the animal is killed without ever gaining consciousness) or required recovery. For the latter there should be appropriate post-surgical care and analgesia, which the paper must also describe. Researchers have an obligation to ensure that pain or distress, whether physical or psychological, is minimised, and steps taken to achieve this should be noted. Describe how the depth of anaesthesia was determined and maintained. This is especially important when neuromuscular blocking agents were employed. Please see below for specific details on the use of neuromuscular blockers. Please note that ether, chloroform and chloral hydrate are not acceptable anaesthetics as they no longer comply with the principles of best practice.

Analgesia and post-operative care in recovery

For animals in recovery there should be appropriate post-surgical care and analgesia, which the paper must also describe. Researchers have an obligation to ensure that pain or distress, whether physical or psychological, is minimised, and steps taken to achieve this should be noted.

Surgical procedures

Provide a brief description of the operation.

Terminal procedures

State what happened to the animals at the end of the study and how they were killed. Animals should have been humanely killed unless reuse in another study had been specifically authorized by the ethics body. Reuse depends upon full restoration of health and well-being and on the severity of the previous and subsequent procedures, evaluated as part of an assessment of the animal’s lifetime experience.

Confirmation of compliance

Include in the Methods section a statement that the investigators understand the ethical principles under which the journal operates and that their work complies with this animal ethics checklist.

 

Additional information on the use of neuromuscular blocking agents and of nitrous oxide

In experiments involving the use of neuromuscular blocking agents authors must describe the precautions taken to ensure the adequacy of anaesthesia. They must provide sufficient detail to enable the reader to determine that no unnecessary suffering occurred. The Physiological Society has issued the following advice:

'When neuromuscular blocking agents are used with anaesthetic agents during physiological experiments, safeguards are required to ensure that the animal does not experience unnecessary pain or distress. It is the responsibility of the person conducting the experiment to ensure that anaesthesia is adequate. Neuromuscular blocking agents should never be used without anaesthesia.

1. For any experiment using neuromuscular blocking agents it should first be established that the proposed anaesthetic regime is adequate, in the absence of these agents, to provide analgesia for any surgical procedure or noxious stimulus which is proposed. When light levels of anaesthesia are considered appropriate for experimental purposes, it should have been established that deeper levels of anaesthesia would interfere with the purpose of the experiment. All preparatory major surgery should be performed under full surgical anaesthesia. Subsequent procedures under light anaesthesia in the presence of neuromuscular blocking agents should be conducted in such a way that any residual pain from the initial surgery is blocked by local anaesthetics or analgesia and no further noxious stimuli are delivered. It should be noted that some methods of head holding using ear bars and zygomatic bars are a potential source of pain: other, atraumatic, methods of head restraint should be used in lightly anaesthetised animals.

2. During the course of each experiment in the period when neuromuscular blocking agents are used there must be a protocol for continuous or regular assessment of adequacy of anaesthesia. The appropriate methods of assessment will depend on the particular anaesthetic and the particular experiment. For example:

(a) the monitoring of changes in heart rate and blood pressure provides one of the most valuable indices of the level of anaesthesia. Neuromuscular blocking agents, in doses which do not reduce blood pressure, do not abolish autonomic cardiovascular reflexes. A preparation in which precipitate cardiovascular responses occur to minor noxious stimuli must be considered too lightly anaesthetised;
(b) the electroencephalogram can be monitored and changes in pattern with minor noxious stimuli may be a valuable guide. Thus, many experiments can be conducted while the EEG is in the unaroused state characteristic of moderate anaesthesia. However, some anaesthetics (e.g. chloralose) induce an EEG that cannot readily be interpreted, and the relation between EEG waveform and the suffering of pain is, in any case, not known. It should also be noted that changes in arterial Pco2 and the administration of atropine and certain other drugs may alter the relation between the EEG and behavioural state;
(c) the state of the pupil can provide a further indication of the level of anaesthesia. Generally, under anaesthesia pupils are constricted, and dilated pupils, or pupils which react rapidly to stimuli, are indicative of inadequate anaesthesia. This is not a suitable test for experiments on the visual system when drugs have been administered which paralyse accommodation and dilate the pupils.

3. Nitrous oxide (N2O) has effective mood changing (tranquillizing) and analgesic properties and is useful because of the ease and consistency of delivery but, even at the maximum concentration feasible at normal pressures, it is not an adequate anaesthetic for surgery in cats. Caution should be used in relying upon it for maintenance, even when precautions have been taken to avoid noxious stimuli. It becomes especially important to assess the animal's state periodically and to use supplementary agents as required.'

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