Skip to main content
NC3Rs: National Centre for the Replacement Refinement & Reduction of Animals in Research
Project grant

The detection, assessment and alleviation of pain in laboratory zebrafish

Zebrafish in multiple tanks

At a glance

Award date
November 2012 - April 2016
Grant amount
Principal investigator
Dr Lynne Sneddon
University of Liverpool


  • Refinement
Read the abstract
View the grant profile on GtR

Application abstract

The use of fish as experimental models has resulted in a significant increase in the numbers used in licensed procedures in the UK (+23% in 2010). Fish are subject to many routine procedures that result in tissue damage that would give rise to pain in mammals. For example, the removal of tissue in fin clipping for identification purposes and genetic screens; invasive tagging where tags are introduced into the body cavity; scraping of skin for disease screening; and invasive surgery with recovery.

This project will identify robust behavioural and physiological indicators to detect and assess pain in a model species, the zebrafish, and use the data collected to develop a simple, cost effective monitoring tool. Video recordings of fish subject to four potentially painful procedures shall be compared with undisturbed and sham treated fish along with non-invasive physiological measures of stress (ventilation rate and water cortisol). An intelligent monitoring system will be produced that can classify individuals according to behavioural measures and will provide an unequivocal means of determining whether fish are behaving normally, are stressed or experiencing pain.

This project will also determine what preferences zebrafish have for physical enrichments and social context using a choice chamber paradigm. Once the top preferences are identified these shall be investigated as a means of improving welfare and the recovery from pain by comparison with fish held under standard, barren conditions. Finally, a range of immersion analgesic and local anaesthetic drugs shall be tested to examine whether they effectively reduce the indicators of pain in zebrafish and what the effective dose is.



  1. Deakin AG et al. (2019). Automated monitoring of behaviour in zebrafish after invasive procedures. Scientific Reports 9(1):9042. doi: 10.1038/s41598-019-45464-w
  2. Deakin A et al. (2019). Welfare Challenges Influence the Complexity of Movement: Fractal Analysis of Behaviour in Zebrafish. Fishes 4(1):8. doi: 10.3390/fishes4010008
  3. Thomson JS et al. (2019) Assessment of behaviour in groups of zebrafish (Danio rerio) using an intelligent software monitoring tool, the chromatic fish analyser. Journal of Neuroscience Methods 328:108433. doi: 10.1016/j.jneumeth.2019.108433
  4. Deakin AG et al. (2019). Welfare challenges influence the complexity of movement: Fractal analysis of behaviour in zebrafish. Fishes 4(1):8. doi: 10.3390/fishes4010008
  5. Sneddon LU (2018). Comparative Physiology of Nociception and Pain. Physiology (Bethesda) 33(1):63-73. doi: 10.1152/physiol.00022.2017
  6. Sneddon LU (2018). Editorial: Where to draw the line? Should the age of protection for zebrafish be lowered? ATLA 46(6):309-311. doi: 10.1177/026119291804600605
  7. Lopez-Luna J et al. (2017). Impact of stress, fear and anxiety on the nociceptive responses of larval zebrafish. PLoS ONE 12(8):e0181010. doi: 10.1371/journal.pone.0181010
  8. Lopez-Luna J et al. (2017). Reduction in activity by noxious chemical stimulation is ameliorated by immersion in analgesic drugs in zebrafish. Journal of Experimental Biology 220(17):3192-3194. doi: 10.1242/jeb.146969
  9. Pounder KC et al. (2017). Physiological and behavioural evaluation of common anaesthesia practices in the rainbow trout. Applied Animal Behaviour Science 199:94-102. doi: 10.1016/j.applanim.2017.10.014
  10. Sneddon LU et al. (2017). Considering aspects of the 3Rs principles within experimental animal biology. Journal of Experimental Biology 220(17):3007-3016. doi: 10.1242/jeb.147058
  11. Sneddon LU (2017). Pain in laboratory animals: A possible confounding factor? ATLA 45(3):161-164. doi: 10.1177/026119291704500309
  12. Osborne N et al. (2016). Report of a Meeting on Contemporary Topics in Zebrafish Husbandry and Care. Zebrafish 13(6):584-589. doi: 10.1089/zeb.2016.1324
  13. Pounder KC et al. (2016). Does environmental enrichment promote recovery from stress in rainbow trout? Applied Animal Behaviour Science 176:136-142. doi: 10.1016/j.applanim.2016.01.009
  14. Sneddon LU (2015). Pain in aquatic animals. Journal of Experimental Biology 218(Pt 7):967-976. doi: 10.1242/jeb.088823 
  15. Schroeder P et al. (2014). What do zebrafish want? Impact of social grouping, dominance and gender on preference for enrichment. Laboratory Animals 48(4):328-37. doi: 10.1177/0023677214538239 
  16. Sneddon LU et al. (2014). Defining and assessing animal pain. Animal Behaviour 97:201-212. doi: 10.1016/j.anbehav.2014.09.007