Considerations for a European animal welfare standard to evaluate adverse phenotypes in teleost fish

Bert, Bettina; Chmielewska, Justyna; Bergmann, Sven; Busch, Maximilian A.; Driever, Wolfgang; Finger‐Baier, Karin; Hößler, Johanna C.; Köhler, Almut; Leich, Nora; Misgeld, Thomas; Nöldner, Torsten; Reiher, Annegret; Schartl, Manfred; Seebach‐Sproedt, Anja; Thumberger, Thomas; Schönfelder, Gilbert; Grune, Barbara

doi:10.15252/embj.201694448

Cited by 24 publications

(18 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1, Nr. 1) in accordance with European Union animal welfare guidelines [25]. The fish facility is under the supervision of the local representative of the animal welfare agency.…”

Section: Methodsmentioning

confidence: 99%

Swift Large-scale Examination of Directed Genome Editing

et al. 2019

Self Cite

View full text Add to dashboard Cite

In the era of CRISPR gene editing and genetic screening, there is an increasing demand for quick and reliable nucleic acid extraction pipelines for rapid genotyping of large and diverse sample sets. Despite continuous improvements of current workflows, the handling-time and material costs per sample remain major limiting factors. Here we present a robust method for low-cost DIY-pipet tips addressing these needs; i.e. using a cellulose filter disc inserted into a regular pipet tip. These filter-in-tips allow for a rapid, stand-alone four-step genotyping workflow by simply binding the DNA contained in the primary lysate to the cellulose filter, washing it in water and eluting it directly into the buffer for the downstream application (e.g. PCR). This drastically cuts down processing time to maximum 30 seconds per sample, with the potential for parallelizing and automation. We show the ease and sensitivity of our procedure by genotyping genetically modified medaka ( Oryzias latipes ) and zebrafish ( Danio rerio ) embryos (targeted by CRISPR/Cas9 knock-out and knock-in) in a 96-well plate format. The robust isolation and detection of multiple alleles of various abundancies in a mosaic genetic background allows phenotype-genotype correlation already in the injected generation, demonstrating the reliability and sensitivity of the protocol. Our method is applicable across kingdoms to samples ranging from cells to tissues i. e. plant seedlings, adult flies, mouse cell culture and tissue as well as adult fish fin-clips.

show abstract

“…1, Nr. 1) in accordance with European Union animal welfare guidelines [25]. The fish facility is under the supervision of the local representative of the animal welfare agency.…”

Section: Methodsmentioning

confidence: 99%

Swift Large-scale Examination of Directed Genome Editing

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…This generates a significant load of paperwork for scientists, for institutions and for regulatory authorities. Furthermore, after the fish have been generated, they must undergo a severity assessment to determine whether they suffer as a result of the genetic modification . Importantly, when the results of this assessment for a given genetically altered line have been published or are available through a repository, the assessment does not have to be carried out again when the line is transferred to a different institution.…”

Section: Annual Number Of Pubmed‐listed Articles Using Zebrafishmentioning

confidence: 99%

Archiving of zebrafish lines can reduce animal experiments in biomedical research

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Fish are considered useful alternatives to mammals within the principles of replacement, reduction and refinement (‘3Rs’) for the use of animals in research. Historically, research into fish welfare has not received much attention and has lagged behind studies in mammals 1 , 7 , 8 although the situation has improved in recent years 9 – 11 . Fish are also subject to the 3Rs principles.…”

Section: Introductionmentioning

confidence: 99%

“…Many laboratory studies use small-bodied model fish species such as the zebrafish Danio rerio and the three-spined stickleback Gasterosteus aculeatus . Advantages of these species include their small size, ease of maintenance in the laboratory and similarities to other vertebrates 9 , 12 – 15 . DNA is frequently collected from these animals to facilitate identification by genotyping, and this is usually achieved by fin clipping under non-terminal anaesthesia 16 .…”

Section: Introductionmentioning

confidence: 99%

Skin swabbing is a refined technique to collect DNA from model fish species

Tilley

Gutiérrez

Sebire

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Model fish species such as sticklebacks and zebrafish are frequently used in studies that require DNA to be collected from live animals. This is typically achieved by fin clipping, a procedure that is simple and reliable to perform but that can harm fish. An alternative procedure to sample DNA involves swabbing the skin to collect mucus and epithelial cells. Although swabbing appears to be less invasive than fin clipping, it still requires fish to be netted, held in air and handled—procedures that can cause stress. In this study we combine behavioural and physiological analyses to investigate changes in gene expression, behaviour and welfare after fin clipping and swabbing. Swabbing led to a smaller change in cortisol release and behaviour on the first day of analysis compared to fin clipping. It also led to less variability in data suggesting that fewer animals need to be measured after using this technique. However, swabbing triggered some longer term changes in zebrafish behaviour suggesting a delayed response to sample collection. Skin swabbing does not require the use of anaesthetics and triggers fewer changes in behaviour and physiology than fin clipping. It is therefore a more refined technique for DNA collection with the potential to improve fish health and welfare.

show abstract

Considerations for a European animal welfare standard to evaluate adverse phenotypes in teleost fish

Cited by 24 publications

References 12 publications

Swift Large-scale Examination of Directed Genome Editing

Swift Large-scale Examination of Directed Genome Editing

Archiving of zebrafish lines can reduce animal experiments in biomedical research

Skin swabbing is a refined technique to collect DNA from model fish species

Contact Info

Product

Resources

About