The number of fish used in research has increased in the last decades. Anaesthesia is required when fish must be held immobile and it is crucial to promote fish welfare, because these vertebrates can show signs of stress and/or pain during handling, transport, tagging, sampling and invasive procedures. The use of an inadequate anaesthetic protocol can compromise not only the welfare of the fish, but also the reliability of the research results. Thus, the development of suitable anaesthetic regimes for each fish species is important. This article reviews the main anaesthetic and analgesic agents used in adult fish in a research setting.

Anaesthesia is used daily in fish experimental procedures; however, the use of an inadequate anaesthetic protocol can compromise not only the animal's welfare but also the reliability of results. The use of zebrafish (Danio rerio) in biomedical research has increased in the last decades, highlighting the importance of appropriate anaesthetic regimens for this species. This article reviews the main anaesthetic agents and protocols used in laboratory adult zebrafish, and some of the analgesic methods to be used in this species that still need more research. In addition, a systematized observation of signs is proposed to evaluate adult zebrafish welfare to reduce pain and distress.

Recent empirical studies have reported evidence that many aquatic species, including fish, cephalopods and crustaceans, have the capacity for nociception and pain, and that their welfare should be taken into consideration. Some sceptics, rejecting the precautionary principle, have denied that any study demonstrates pain or other aspects of sentience in fish. This target article discusses some of the scientific shortcomings of these critiques through a detailed analysis of a study exploring nociception and analgesia in larval zebrafish.

The increasing use of zebrafish model has not been accompanied by the evolution of proper anaesthesia for this species in research. The most used anaesthetic in fishes, MS222, may induce aversion, reduction of heart rate, and consequently high mortality, especially during long exposures. Therefore, we aim to explore new anaesthetic protocols to be used in zebrafish by studying the quality of anaesthesia and recovery induced by different concentrations of propofol alone and in combination with different concentrations of lidocaine. In experiment A, eighty-three AB zebrafish were randomly assigned to 7 different groups: control, 2.5 (2.5P), 5 (5P) or 7.5 μg/ml (7.5P) of propofol; and 2.5 μg/ml of propofol combined with 50, (P/50L), 100 (P/100L) or 150 μg/ml (P/150L) of lidocaine. Zebrafish were placed in an anaesthetic water bath and time to lose the equilibrium, reflex to touch, reflex to a tail pinch, and respiratory rate were measured. Time to gain equilibrium was also assessed in a clean tank. Five and 24 hours after anaesthesia recovery, zebrafish were evaluated concerning activity and reactivity. Afterwards, in a second phase of experiments (experiment B), the best protocol of the experiment A was compared with a new group of 8 fishes treated with 100 mg/L of MS222 (100M). In experiment A, only different concentrations of propofol/lidocaine combination induced full anaesthesia in all animals. Thus only these groups were compared with a standard dose of MS222 in experiment B. Propofol/lidocaine induced a quicker loss of equilibrium, and loss of response to light and painful stimuli compared with MS222. However zebrafish treated with MS222 recovered quickly than the ones treated with propofol/lidocaine. In conclusion, propofol/lidocaine combination and MS222 have advantages in different situations. MS222 is ideal for minor procedures when a quick recovery is important, while propofol/lidocaine is best to induce a quick and complete anaesthesia.