Anaesthesia of laboratory, aquaculture and ornamental fish: Proceedings of the first LASA-FVS Symposium.

Following on from the Annual Fish Veterinary Society Conference, this symposium was organised with the Laboratory Animal Science Association and brought together experts from ornamental (pond and aquarium) fish practice, aquaculture and aquatic-research facilities to discuss good practice of anaesthesia. This proceedings paper gives an overview of relevant experiences involving a range of immersion drugs including tricaine, benzocaine and isoeugenol, as well as a summary of the main topics of discussion. While fish anaesthesia is commonplace, administration methods, drugs and monitoring procedures may often be regarded as antiquated when compared with mammalian practice. These limitations notwithstanding, individual fish will benefit from good anaesthetic monitoring. Although the most common anaesthetic drugs may be perceived as equally efficacious and therefore interchangeable for different settings, challenges are different for the anaesthesia of grouped fish, when determining species-dependent anaesthetic dosing in a multi-species tank, or adapting to farming requirements, nationally licensed products, costs and withdrawal periods. The fish anaesthetic arsenal fails to address premedication, analgesia and issues of averseness. The two latter factors should be part of the evaluation of anaesthetic protocols; therefore, instructions for the analgesic provision of lidocaine to fin clipped zebrafish are proposed. Euthanasia practices could sometimes be refined too. Alternative physical methods such as electrical stunning are options to be considered.

Species specific anaesthetics for fish anaesthesia and euthanasia.

There is a need to ensure that the care and welfare for fish maintained in the laboratory are to the highest standards. This extends to the use of anaesthetics for both scientific study, humane killing and euthanasia at end of life. An anaesthetic should not induce negative behaviours and fish should not seek to avoid the anaesthetic. Surprisingly little information is available to facilitate a humane choice of anaesthetic agent for fish despite over 100 years of use and the millions of fish currently held in thousands of laboratories worldwide. Using a chemotaxic choice chamber we found different species specific behavioural responses among four closely related fish species commonly held in the laboratory, exposed to three widely used anaesthetic agents. As previously found for zebrafish (Danio rerio), the use of MS-222 and benzocaine also appears to induce avoidance behaviours in medaka (Oryzias latipes); but etomidate could provide an alternative choice. Carp (Cyprinus carpio), although closely related to zebrafish showed avoidance behaviours to etomidate, but not benzocaine or MS-222; and rainbow trout (Oncorhynchus mykiss) showed no avoidance to the three agents tested. We were unable to ascertain avoidance responses in fathead minnows (Pimephales promelas) and suggest different test paradigms are required for that species.

A Good Death? Report of the Second Newcastle Meeting on Laboratory Animal Euthanasia.

Millions of laboratory animals are killed each year worldwide. There is an ethical, and in many countries also a legal, imperative to ensure those deaths cause minimal suffering. However, there is a lack of consensus regarding what methods of killing are humane for many species and stages of development. In 2013, an international group of researchers and stakeholders met at Newcastle University, United Kingdom to discuss the latest research and which methods could currently be considered most humane for the most commonly used laboratory species (mice, rats and zebrafish). They also discussed factors to consider when making decisions about appropriate techniques for particular species and projects, and priorities for further research. This report summarises the research findings and discussions, with recommendations to help inform good practice for humane killing.

Do fish perceive anaesthetics as aversive?

This study addresses a fundamental question in fish welfare: are the anaesthetics used for fish aversive? Despite years of routine general use of many agents, within both scientific research and aquaculture, there is a paucity of information regarding their tolerance and associated behavioural responses by fish. This study examined nine of the most commonly used fish anaesthetic agents, and performed preference tests using adult mixed sex zebrafish (Danio rerio), the most commonly held laboratory fish. Video tracking software quantified swimming behaviour related to aversion for each anaesthetic at 50% of its standard recommended dose compared with clean water in a flow-through chemotaxic choice chamber. Results suggest that several commonly used anaesthetics were aversive, including two of the most commonly recommended and used: MS222 (ethyl 3-aminobenzoate methanesulphate) and benzocaine. For ethical best practice, it is recommended that compounds that are aversive, even at low concentration, should no longer be used routinely for anaesthesia or indeed the first step of humane euthanasia of adult zebrafish. Two agents were found not to induce aversive behavioural responses: etomidate and 2,2,2 tribromoethanol. For the millions of adult zebrafish used in laboratories and breeding worldwide, etomidate appears best suited for future routine humane use.