A Low-Cost Method of Skin Swabbing for the Collection of DNA Samples from Small Laboratory Fish

Breacker, Carl; Barber, Iain; Norton, William; McDearmid, Jonathan R.; Tilley, Ceinwen A.

doi:10.1089/zeb.2016.1348

Cited by 42 publications

(50 citation statements)

References 26 publications

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“…rerio exposed to anaesthetics. The authors suggest that fin‐clipping is an area that could offer the opportunity for refinement if genotyping might be completed without the invasive fin‐clip and recently this has been reported (Breacker et al ., ). There are efforts reported in the scientific community to refine this procedure in other species such as Neolamprologus pulcher (Trewavas & Poll 1952) (Le Vin et al ., ) and Gasterosteus aculeatus L. 1758 (Sebire et al ., ).…”

Section: Discussionmentioning

confidence: 99%

International survey on the use and welfare of zebrafish Danio rerio in research

Lidster

Readman

Prescott

et al. 2017

Journal of Fish Biology

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A survey was conducted regarding zebrafish Danio rerio use for scientific research with a focus on: anaesthesia and euthanasia; housing and husbandry; breeding and production; refinement opportunities. A total of 98 survey responses were received from laboratories in 22 countries in Europe, North America, South America, Asia and Australia. There appears a clear and urgent need to identify the most humane methods of anaesthesia and euthanasia. Aversive responses to MS-222 were widely observed raising concerns about the use of this anaesthetic for D. rerio. The use of anaesthesia in fin clipping for genetic identification is widely practised and there appears to be an opportunity to further develop less invasive methods and refine this process. Optimization (and potentially standardization) of feeding is an area for further investigation. Given that diet and body condition can have such profound effects on results of experiments, differences in practice could have significant scientific implications. Further research into transition between dark and light phases in the laboratory appears to represent an opportunity to establish best practice. Plants and gravel were not considered practical by many laboratories. The true value and benefits need to be established and communicated. Overproduction is a concern both from ethical and financial viewpoints. There is an opportunity to further reduce wastage of D. rerio. There are clear concerns and opportunities for the scientific community to work together to further improve the welfare of these important laboratory models.

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Section: Discussionmentioning

confidence: 99%

International survey on the use and welfare of zebrafish Danio rerio in research

Lidster

Readman

Prescott

et al. 2017

Journal of Fish Biology

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“…We determined the sex of each individual using a genetic sex-linked marker (Peichel et al, 2004) prior to exposure, using a non-invasive skin swabbing and DNA extraction method (Breacker et al, 2016).…”

Section: Sex Identificationmentioning

confidence: 99%

Can the behaviour of threespine stickleback parasitized with Schistocephalus solidus be replicated by manipulating host physiology?

Hébert

Berger

Barber

et al. 2016

Journal of Experimental Biology

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Sticklebacks infected by the parasitic flatworm Schistocephalus solidus show dramatic changes in phenotype, including a loss of species-typical behavioural responses to predators. The timing of host behaviour change coincides with the development of infectivity of the parasite to the final host (a piscivorous bird), making it an ideal model for studying the mechanisms of infection-induced behavioural modification. However, whether the loss of host anti-predator behaviour results from direct manipulation by the parasite, or is a by-product (e.g. host immune response) or side effect of infection (e.g. energetic loss), remains controversial. To understand the physiological mechanisms that generate these behavioural changes, we quantified the behavioural profiles of experimentally infected fish and attempted to replicate these in non-parasitized fish by exposing them to treatments including immunity activation and fasting, or by pharmacologically inhibiting the stress axis. All fish were screened for the following behaviours: activity, water depth preference, sociability, phototaxis, anti-predator response and latency to feed. We were able to change individual behaviours with certain treatments. Our results suggest that the impact of S. solidus on the stickleback might be of a multifactorial nature. The behaviour changes observed in infected fish might result from the combined effects of modifying the serotonergic axis, lack of energy and activation of the immune system.

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“…For Kca33Tg and Kca66Tg lines we used the F2 progeny of founders obtained from ZIRC. DNA was obtained by "scale swabs" from fish present in the laboratory using a modified protocol of previously established methods 19 . Briefly, unanesthetized fish are removed from the tank and placed on a wet sponge or gauze.…”

Section: Methodsmentioning

confidence: 99%

A 13-plex of tetra- and penta-STRs to identify zebrafish

Venta

Nguyen

Senut

et al. 2020

Sci Rep

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The zebrafish species Danio rerio has become one of the major vertebrate model organisms used in biomedical research. However, there are aspects of the model that need to be improved. One of these is the ability to identify individual fish and fish lines by DNA profiling. Although many dinucleotide short tandem repeat (diSTR) markers are available for this and similar purposes, they have certain disadvantages such as an excessive polymerase slippage ("stutter") that causes difficulties in automated genotyping and cross-laboratory comparisons. Here we report on the development of a 13-plex of tetranucleotide and pentanucleotide STRs (tetraSTRs and pentaSTRs, respectively) that have low stutter. The system uses an inexpensive universal primer labelling system, which can easily be converted to a direct labeling system if desired. This 13-plex was examined in three zebrafish lines (NHGRI-1, kca33Tg, and kca66Tg, originally obtained from ZIRC). The average observed heterozygosity (Ho) and expected heterozygosity (He) in these highly inbred lines were 0.291 and 0.359, respectively, which is very similar to what has been found with diSTRs. The probability of identity (PI) for all fish tested was 2.1 × 10 −5 and the PI for siblings (PIsib) was 6.4 × 10 −3 , as calculated by the Genalex package. Ninety percent of the fish tested were correctly identified with their respective strains. It is also demonstrated that this panel can be used to confirm doubled-haploid cell lines. This multiplex should find multiple uses for improving the accuracy and reproducibility of studies using the zebrafish model. Zebrafish (Danio rerio) is the vertebrate animal model most rapidly adopted in biomedical research. The zebrafish genome is 70% similar to the human, and 80% percent of the genes responsible for human diseases have an orthologue in zebrafish 1. It is relatively easy to generate mutant animals, and the cost of zebrafish maintenance is low. Applications of this animal model include, among the many, toxicology, developmental biology, genetics and degenerative diseases, psychiatric conditions, cancer, and metabolic disorders. Researchers can choose from a variety of wild-type and mutant zebrafish lines for their studies. All available from the Zebrafish International Resource Center (ZIRC), wild-type zebrafish lines include Tübingen, AB, NHGRI-1, Sanger AB Tübingen (SAT), WIK, Tüpfel long fin (TL), Nadia, and Cooch Behar. Also available, are more than forty thousand mutant lines. Despite previous reports of the derivation of the IM inbred zebrafish line in 2011, homozygous zebrafish lines for all loci are lacking 2,3. Some initiatives are in progress that may fulfill the need for fish lines that have an identical genetic identity. We are currently working on a strategy to generate homozygous diploid fish using the method developed by Streisinger and colleagues in 1981, followed by somatic cell nuclear transfer (SCNT) 4-9. Once a gynogenetic embryo is developed, we isolate cells from them, and subsequently, after testing for normal k...

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A Low-Cost Method of Skin Swabbing for the Collection of DNA Samples from Small Laboratory Fish

Cited by 42 publications

References 26 publications

International survey on the use and welfare of zebrafish Danio rerio in research

International survey on the use and welfare of zebrafish Danio rerio in research

Can the behaviour of threespine stickleback parasitized with Schistocephalus solidus be replicated by manipulating host physiology?

A 13-plex of tetra- and penta-STRs to identify zebrafish

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