Impact of analgesic drugs on the behavioural responses of larval zebrafish to potentially noxious temperatures

López-Luna, Javier; Al-Jubouri, Qussay; Al-Nuaimy, Waleed; Sneddon, Lynne U.

doi:10.1016/j.applanim.2017.01.002

Cited by 78 publications

(56 citation statements)

References 42 publications

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“…Diggles et al also fail to cite a similar study that clearly undermines their position that altered water quality explains the behavioural changes (Lopez-Luna et al, 2017b). This study used heat as a noxious stimulus rather than acid.…”

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confidence: 94%

Responses of larval zebrafish to low pH immersion assay. Comment on Lopez-Luna et al.

Diggles

Arlinghaus

Browman

et al. 2017

Journal of Experimental Biology

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Lopez -Luna et al. (2017) observed behavioral responses of larval zebrafish (Danio rerio) exposed for 10 min to pH 2.6-3.6 when acetic acid (0.01-0.25%) or citric acid (0.1-5%) was added to the tank water in the presence or absence of aspirin (1-2.5 mg l −1 ), morphine sulfate (1-48 mg l −1 ), lidocaine (1-5 mg l −1 ) and flunixin (8-20 mg l −1 ). Fish exposed to 0.1-0.25% acetic acid were less active than controls while those exposed to citric acid and 0.01% acetic acid were more active. Administration of high doses of aspirin, morphine and lidocaine for 30 min before exposure prevented the reduction in activity induced by 0.1-0.25% acetic acid.These behavioral responses were interpreted as evidence that acetic acid immersion provided a noxious stimulus (i.e. activated nociceptors) that was reliable for use as a model system for the study of analgesic substances. We identify methodological weaknesses and inconsistencies in the interpretation of results, and emphasize that activation of nociceptors was assumed, not demonstrated. As a result of several processes and interactions that were not accounted for or discussed, we warn their conclusions are unfounded.A critical omission was the failure to report water conductivity, hardness and alkalinity data. These determine the magnitude of acute osmoregulatory effects that occur in fish exposed to highly acidic water . Trials by other researchers using water with different conductivity, hardness or alkalinity profiles could, therefore, generate significantly different results. Immersion of fish in low pH water also introduces several unavoidable and uncontrolled interactions that prevent unequivocal interpretation of the behavioral changes observed.For example, sudden exposure of fish to water of pH <4 results in gill dysfunction, iono-regulatory failure and pathological lesions of the gill epithelium . These reduce respiratory efficiency, initiating compensatory behavioral responses such as surface respiration (Kramer, 1987), which appears synonymous with 'top-dwelling behavior' reported by Currie (2014) in adult zebrafish immersed in 0.03% acetic acid ( pH 3.9-4.0). Notably, aquatic surface respiration can occur in a variety of natural circumstances in the absence of nociception (Kramer, 1987), so this behavior is insufficient evidence that nociception is occurring.In contrast to Currie (2014) and Steenbergen and Bardine (2014), Lopez-Luna et al. (2017) considered reduced (not increased) activity as evidence of 'alleged pain behavior' in zebrafish exposed to 0.1-0.25% acetic acid. Steenbergen and Bardine (2014) interpreted increased activity and cyclooxygenase-2 gene expression as evidence of nociception in larval zebrafish immersed in 0.0025-0.025% acetic acid. However, cyclooxygenase-2 expression is a non-specific marker of several physiological processes (Wang et al., 2016), meaning its expression is also insufficient evidence of nociception. A critical observation is that larval zebrafish in the study by Lopez-Luna et al. (2017) continued to exhibit increased a...

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confidence: 94%

Responses of larval zebrafish to low pH immersion assay. Comment on Lopez-Luna et al.

Diggles

Arlinghaus

Browman

et al. 2017

Journal of Experimental Biology

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“…Empirical studies in fishes have also confirmed the presence of nociceptors, demonstrating their properties are very similar to those found in humans and other mammals (Ashley et al ., , ; Mettam et al ., ; Sneddon, , , ). Studies in fishes have demonstrated changes in brain activity during noxious stimuli (Dunlop & Laming, ; Nordgreen et al ., ; Reilly et al ., ; Sneddon, ) and recorded adverse changes in behaviour and physiology during noxious treatments (Dunlop et al ., ; Maximino, ; Mettam et al ., ; Reilly et al ., ; Roques et al ., ; Sneddon, ; Sneddon et al ., , ; White et al ., ) all of which can be prevented by analgesics known to be effective in relieving pain in mammals (Lopez‐Luna et al ., , , , ; Mettam et al ., ; Newby et al ., ; Nordgreen et al ., ; Schroeder & Sneddon, ; Sneddon, , ; Taylor et al ., ). This empirical evidence taken together supports the idea that fishes may experience pain.…”

Section: Do Fishes Experience Pain?mentioning

confidence: 99%

“…Opioids inhibit neurotransmitter release pre‐synaptically thereby blocking nociceptors but these drugs also block central transmission. Several studies have shown morphine is an effective analgesic in O. mykiss (Baker et al ., ; Jones et al ., ; Sneddon, ) and other species ( e.g ., goldfish Carassius auratus (L. 1758); Nordgreen et al ., ; D. rerio , Lopez‐Luna et al ., ,b,c,d; Magalhaes et al ., ; Schroeder & Sneddon, ; Taylor et al ., ). Morphine has a pharmocokinetically similar action in fishes compared with mammals, however, morphine persists for a prolonged period after administration due to slower excretion rates (half‐life 37 h; Newby et al ., , ).…”

Section: Common Considerations For Laboratory and Field Studiesmentioning

confidence: 99%

“…Indomethacin injected intraperitoneally (20 μl of 0.2 mg ml −1 ) did reduce pain related responses in adult D. rerio treated with formalin (Magalhaes et al ., ). Aspirin administered via immersion prevented the reduced activity seen in adult and larval D. rerio when subjected to noxious events (Lopez‐Luna et al ., ,b,c,d; Schroeder & Sneddon, ). Future studies need to investigate NSAIDs further to determine the minimum effective dose and explore any unwanted side‐effects since long‐term use of these drugs in mammals, birds and reptiles can result in gastric ulcers or renal disease.…”

Section: Common Considerations For Laboratory and Field Studiesmentioning

confidence: 99%

“…Lidocaine effectively prevents behavioural and physiological alterations in O. mykiss during subcutaneous injection of acetic acid (Mettam et al ., ). Exposure to lidocaine by emersion also ameliorates substantial changes in the behaviour of adult D. rerio subject to caudal‐fin clipping (Schroeder & Sneddon, ; Deakin et al ., ) and was also effective when used with larval D. rerio exposed to noxious chemical and thermal stimuli (Lopez‐Luna et al ., ,b,c,d). Infiltration of rainbow trout with lidocaine appears to damage skeletal muscle and connective tissue with higher concentrations of (10 mg kg −1 ) but not to cause adverse behavioural side‐effects (20 mg kg −1 ), compared with saline infiltration (Chatigny et al ., 2018a,b).…”

Section: Common Considerations For Laboratory and Field Studiesmentioning

confidence: 99%

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Ethical considerations in fish research

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Bouyoucos

Brooks

et al. 2019

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Fishes are used in a wide range of scientific studies, from conservation research with potential benefits to the species used to biomedical research with potential human benefits. Fish research can take place in both laboratories and field environments and methods used represent a continuum from non‐invasive observations, handling, through to experimental manipulation. While some countries have legislation or guidance regarding the use of fish in research, many do not and there exists a diversity of scientific opinions on the sentience of fish and how we determine welfare. Nevertheless, there is a growing pressure on the scientific community to take more responsibility for the animals they work with through maximising the benefits of their research to humans or animals while minimising welfare or survival costs to their study animals. In this review, we focus primarily on the refinement of common methods used in fish research based on emerging knowledge with the aim of improving the welfare of fish used in scientific studies. We consider the use of anaesthetics and analgesics and how we mark individuals for identification purposes. We highlight the main ethical concerns facing researchers in both laboratory and field environments and identify areas that need urgent future research. We hope that this review will help inform those who wish to refine their ethical practices and stimulate thought among fish researchers for further avenues of refinement. Improved ethics and welfare of fishes will inevitably lead to increased scientific rigour and is in the best interests of both fishes and scientists.

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Zebrafish

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2024

The UFAW Handbook on the Care and Management of Laboratory and Other Research Animals

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Impact of analgesic drugs on the behavioural responses of larval zebrafish to potentially noxious temperatures

Cited by 78 publications

References 42 publications

Responses of larval zebrafish to low pH immersion assay. Comment on Lopez-Luna et al.

Responses of larval zebrafish to low pH immersion assay. Comment on Lopez-Luna et al.

Ethical considerations in fish research

Zebrafish

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