Zebrafish

Do you work with zebrafish in the Nordic countries?
Are you interested in a practical course on the care and use of zebrafish?
The Nordic Zebrafish Network was founded in Stockholm on 17 November 2023. Please see this overview of its activities.

Zebrafish (Danio rerio) are being used in increasing numbers within molecular biology, developmental biology, neurobiology, genetics, cancer research and drug discovery, due to the low costs of maintaining them, their short generation interval, the transparency of the embryos and the ability to manipulate the genome.
A revision of the requirements for housing and humane killing of zebrafish (Annexes III & IV of the EU Directive) was published in October 2023.

In addition to the topics below, please note that the PREPARE guidelines have separate tabs for fish researchers (see an example here) on each of the pages describing the topics on the PREPARE checklist.

• General references
• Housing and welfare
• Environmental enrichment
• Health monitoring
• Anaesthesia and analgesia
• Procedures
• Euthanasia
• Other literature and media articles
• Organisations and resource collections
• Meetings
• Zebrafish facilities

General references

• Recordings from the Zebrafish Refinement Webinar organised by Swiss 3RCC, 6 November 2023
• General resources for the care and use of fish in research
• Textbooks on zebrafish
• Zebrafish and other small warm-water laboratory fish (CCAC guidelines)
• Zebrafish welfare page (NC3Rs)
• International survey on the use and welfare of zebrafish Danio rerio in research (Lidster et al., 2017)

Housing and welfare

• Support material for the acclimatisation of zebrafish prior to experiments (Swedish 3Rs-Center, 2024; also in Swedish)
• vom Berg (2023): Refinement of zebrafish husbandry (webinar 6 November)
• Zbinden (2023): Culture of care in zebrafish (webinar 6 November)
• A collection of links to resources for the housing and husbandry of zebrafish, from the NA3RsC
• Welfare Assessment of Adult Laboratory Zebrafish: A Practical Guide (Sabrautzki et al., 2021)
• Effects of holding density on the welfare of zebrafish: A systematic review (Andersson & Kettunen, 2021)
• Different influences of anxiety models, environmental enrichment, standard conditions and intraspecies variation (sex, personality and strain) on stress and quality of life in adult and juvenile zebrafish: A systematic review (Buenhombre et al., 2021)
• Zebrafish (Danio rerio) Environmental Summary, Aquatic Resources Program, Boston Children's Hospital  (Lawrence et al., 2021)
• A fish is not a mouse: understanding differences in background genetics is critical (Crim & Lawrence, 2020)
• Automated monitoring of behaviour in zebrafish after invasive procedures (Deakin et al., 2019)
• Zebrafish: Housing and husbandry recommendations (2019): Joint Working Group Report from the European Society for Fish Models in Biology and Medicine (EUFishBioMed), which has since changed its name to the European Zebrafish Society (EZS) and FELASA
• Guidance on the housing and care of zebrafish (2011), produced by the Animals in Science Department, RSPCA
• Welfare challenges influence the complexity of movement: Fractal analysis of behaviour in zebrafish (Deakin et al., 2019)
• Is heightened-shoaling a good candidate for positive emotional behavior in zebrafish? (Franks et al., 2018)
• The role of water chemistry in zebrafish welfare and reproducibility of research studies (project led by Roderic Wilson, started in 2018)
• What do zebrafish want? Impact of social grouping, dominance and gender on preference for enrichment (Schroeder et al., 2014)
  
  

Environmental enrichment

• Stevens (2023): Implementing enrichment for laboratory zebrafish (webinar 6 November)
• A systematic review of the impact of environmental enrichment in zebrafish (Gallas-Lopes et al., 2023)
• Enrichment for Laboratory Zebrafish - A Review of the Evidence and the Challenges (Stevens et al., 2021)
• Zebrafish environmental enrichment posters from the RSPCA
• Different influences of anxiety models, environmental enrichment, standard conditions and intraspecies variation (sex, personality and strain) on stress and quality of life in adult and juvenile zebrafish: A systematic review (Buenhombre et al., 2021)
• Effects of environmental enrichment on survivorship, growth, sex ratio and behaviour in laboratory maintained zebrafish Danio rerio (Lee et al., 2018)
• What do zebrafish want? Impact of social grouping, dominance and gender on preference for enrichment (Schroeder et al., 2014)

Health monitoring

• A FELASA Working Group Survey on Fish Species Used for Research, Methods of Euthanasia, Health Monitoring, and Biosecurity in Europe, North America, and Oceania (Mocho & von Krogh, 2022).
• Body Condition Scoring For Adult Zebrafish (Danio rerio) (Clark et al., 2018)
• Recommendations for health monitoring and reporting for zebrafish research facilities (Collymore et al., 2016)
• Toward an integrated zebrafish health management program supporting cancer and neuroscience research (Martins et al., 2016)
• Guidelines for health and welfare monitoring of fish in research (Johansen et al., 2006)

Anaesthesia and analgesia
See also the more general section on anaesthesia and analgesia of fish
General principles of anaesthesia and analgesia

• Sneddon (2023): Pain assessment, anaesthesia and analgesia in zebrafish (webinar 6 November)
• FELASA (2023): Pain management in zebrafish: Report from a FELASA Working Group
• Davis et al. (2022): Propofol Immersion As a Euthanasia Method for Adult Zebrafish (Danio Rerio)
• Analgesia, Anesthesia and Euthanasia in zebrafish (Köhler & Valentim, 2021) - chapter 6 in Laboratory Fish in Biomedical Research
• Adult Zebrafish Anesthesia: A Study of Efficacy and Behavioral Recovery of Different Anesthetics (Jorge et al., 2021)
• A suitable anaesthetic protocol for metamorphic zebrafish (Owen & Kelsh, 2021)
• Tricaine Methanesulfonate (MS222) Has Short-Term Effects on Young Adult Zebrafish (Danio rerio) Working Memory and Cognitive Flexibility, but Not on Aging Fish (Fontana et al., 2021)
• Morphine alleviates pain after heart cryoinjury in zebrafish without impeding regeneration (Lelek et al., 2020)
• Anesthesia, Analgesia, and Euthanasia of the Laboratory Zebrafish (Collymore, 2020) - chapter 34 in The Zebrafish in Biomedical Research  
• Screening of Anaesthetics in Adult Zebrafish (Danio rerio) for the Induction of Euthanasia by Overdose (von Krogh et al., 2021)
• Impact of stress, fear and anxiety on the nociceptive responses of larval zebrafish (Lopez-Luna J et al., 2017)
• A Low-Cost Method of Skin Swabbing for the Collection of DNA samples from Small Laboratory Fish (Breacker et al., 2019)
• Automated monitoring of behaviour in zebrafish after invasive procedures Deakin et al. (2019)
• Exploring the efficacy of immersion analgesics in zebrafish using an integrative approach (Schroeder & Sneddon, 2017)
• Anaesthesia and analgesia in zebrafish: a question of refinement (Martins et al., 2016)
• A New Anaesthetic Protocol for Adult Zebrafish (Danio rerio): Propofol combined with Lidocaine (Valentim et al., 2016)
• Conditioned Place Avoidance of Zebrafish (Danio rerio) to Three Chemicals Used for Euthanasia and Anaesthesia (Wong et al., 2014)
• Behavioural Effects of the Commonly Used Fish Anaesthetic Tricaine Methanesulfonate (MS-222) on Zebrafish (Danio rerio) and Its Relevance for the Acetic Acid Pain Test (Nordgreen et al., 2014)
• Efficacy and Safety of 5 Anesthetics in Adult Zebrafish (Danio rerio) (2014) (Collymore et al., 2014)
• Do Fish perceive Anaesthetics as Aversive? (Readman et al., 2013)
• Anesthesia and Euthanasia in Zebrafish (Matthews & Varga, 2012)
• A Novel Behavioral Fish Model of Nociception for Testing Analgesics (Correia et al., 2011)
• Combined use of MS-222 (tricaine) and isofluorane extends anesthesia time and minimizes cardiac rhythm side effects in adult zebrafish Huang et al., 2010)
• Guidelines for anaesthesia and analgesia of fish (Goodman, 2009)
• The Efficacy of Clove Oil as an Anesthetic for the Zebrafish, (Danio rerio) Hamilton (Grush et al., 2004)
  
  Procedures
• Norton (2023): Skin swabbing as a refined technique to collect DNA from laboratory fish (webinar 6 November)
• Hernando (2023): The EggSorter, an automated tool to standardize the processing of zebrafish embryos (webinar 6 November)
• Skin swabbing for DNA sampling of zebrafish (NC3Rs site)
• Skin swabbing is a refined technique to collect DNA from model species (Tilley et al., 2020)
• Automated monitoring of behaviour in zebrafish after invasive procedures (the Fish Behaviour Index) (Deakin et al., 2019)
• A low-cost method of skin swabbing for the collection of DNA samples from small laboratory fish (Breacker et al., 2017)  Powerpoint slide about the technique (von Krogh)
• A Gelatin-based Diet for Oral Dosing Juvenile to Adult Zebrafish (Danio rerio) (Sciarra et al., 2014)
• Guidance on the severity classification of procedures involving fish (Hawkins et al., 2011)
• Dissection of Organs from the Adult Zebrafish (Gupta & Mullins, 2010)
• An overview of existing guidelines for handling, bleeding, administration and identification techniques for fish (Hawkins, 2009)
• An Encyclopedia of Experiments (video collection of research techniques on JOVE)

Euthanasia

• A Multi-Site Assessment of Anesthetic Overdose, Hypothermic Shock, and Electrical Stunning as Methods of Euthanasia for Zebrafish (Danio rerio) Embryos and Larvae (Mocho et al., 2022)
• A FELASA Working Group Survey on Fish Species Used for Research, Methods of Euthanasia, Health Monitoring, and Biosecurity in Europe, North America, and Oceania (Mocho & von Krogh, 2022)
• Report of Workshop on Euthanasia for Zebrafish - A Matter of Welfare and Science (Köhler et al., 2017)
• Euthanizing zebrafish legally in Europe: Are the approved methods of euthanizing zebrafish appropriate to research reality and animal welfare? (Valentim et al., 2016)
• Effectiveness of recommended euthanasia methods in larval zebrafish (Danio rerio) (Strykowski & Schech, 2015)
• Evaluation of rapid cooling and Tricaine Methanesulfonate (MS222) as methods of euthanasia in Zebrafish (Danio rerio) (Wilson et al., 2009)

Other literature and media articles

• Improving zebrafish laboratory welfare and scientific research through understanding their natural history  (Lee et al., 2022)
• Zebrafish (Danio rerio) meets bioethics: the 10Rs ethical principles in research (Canedo et al., 2022)
• Zebrafish: the rising star of animal models (Botthof)
• Behind the scenes of zebrafish research (Botthof)
• Zebrafish Rock! (Twitter/X account)

Organisations and resource collections

• ZFIN Protocol Wiki
• Zebrafish Anatomy Portal (ZFAP)
• The European Zebrafish Society (EZS), formerly EuFishBioMed
• European Zebrafish Resource Center (EZRC)
• Zebrafish Husbandry Association (ZHA)
• Zebrafish Information Network (ZFIN)
• Zebrafish International Resource Center (ZIRC)
• Zebrafish journal (with Technofish sections)
• ANZAAP (Australian and New Zealand Association of Aquarium Professionals)
  
  

Meetings

• Focus on Fish (meetings arranged by the RSPCA in 2021 & 2022), which contained several presentations about zebrafish
• Improved zebrafish welfare through enrichment, behavioural assessment and analgesia (NC3Rs/IAT Symposium 2019)
• Nordic Zebrafish Meeting (2018), Karolinska Institute
• Presentations and Report from the 1st Nordic Zebrafish & Medaka Care Workshop, Oslo, 7 February 2018:
  • Norecopa: Resources for improving animal research (Adrian Smith)
  • Zebrafish Husbandry Association: available resources for the community (Catarina Certal)
  • Zebrafishing in Lisbon (Congento) (Catarina Certal)
  • Can technology in Lab Animal Science support reproducible and precise research? (Massimo Ferrari)
  • Euthanasia of zebrafish and medaka (Kristine von Krogh)
  • Common health problems in lab fish and how to detect them (Marianne Kraugerud)
  • Update from the Joint Working Group for Zebrafish Guidelines (Peter Aleström) 
  • The Zebrafish core facility at Karolinska Institut (Lars Bräutigam)
  • Report from the meeting
• 3Rs Resources and Guidelines for Zebrafish (presentation by Adrian Smith at the NC3Rs/Cefas workshop on Addressing the needs for refinement of laboratory fish in Weymouth, 26-28 April 2016). This presentation includes an overview of international guidelines for the care and use of zebrafish; organisations and journals related to zebrafish research; suggestions on how to improve literature searches and reporting. A shorter version, just containing the 3R resources and guidelines, is available here.
• Detection and alleviation of pain in fish, a presentation by Dr. Lynne Sneddon at Norecopa's annual general meeting in 2015
• Contemporary Topics in Zebrafish Husbandry and Care: report of a meeting in 2014
• Presentations from Harmonisation of the Care and Use of Fish in Research, Gardermoen 2009

Please see Norecopa's International Meetings Calendar and list of past meetings (where there still may be information on the meeting websites).

Zebrafish facilities

• Zebrafish Facility, Bateson Centre, University of Sheffield
• Laboratory of Lynne Sneddon, University of Gothenburg