A nondestructive, rapid, reliable and inexpensive method to sample, store and extract high‐quality DNA from fish body mucus and buccal cells

Lucentini, Livia; Palomba, Antonella; Lancioni, Hovirag; Natali, Mauro; Panara, Fausto

doi:10.1111/j.1471-8286.2005.01142.x

Cited by 49 publications

(35 citation statements)

References 14 publications

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“…Unlike fish and amphibians, which release abundant extracellular DNA via body mucus secretion [17], crayfish and other crustacean species seem to release limited amounts of tissue into water, which makes them more difficult to detect.…”

Section: Introductionmentioning

confidence: 99%

Using eDNA to detect the distribution and density of invasive crayfish in the Honghe-Hani rice terrace World Heritage site

Cai

Yang

et al. 2017

PLoS ONE

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The Honghe-Hani landscape in China is a UNESCO World Natural Heritage site due to the beauty of its thousands of rice terraces, but these structures are in danger from the invasive crayfish Procambarus clarkii. Crayfish dig nest holes, which collapse terrace walls and destroy rice production. Under the current control strategy, farmers self-report crayfish and are issued pesticide, but this strategy is not expected to eradicate the crayfish nor to prevent their spread since farmers are not able to detect small numbers of crayfish. Thus, we tested whether environmental DNA (eDNA) from paddy-water samples could provide a sensitive detection method. In an aquarium experiment, Real-time Quantitative polymerase chain reaction (qPCR) successfully detected crayfish, even at a simulated density of one crayfish per average-sized paddy (with one false negative). In a field test, we tested eDNA and bottle traps against direct counts of crayfish. eDNA successfully detected crayfish in all 25 paddies where crayfish were observed and in none of the 7 paddies where crayfish were absent. Bottle-trapping was successful in only 68% of the crayfish-present paddies. eDNA concentrations also correlated positively with crayfish counts. In sum, these results suggest that single samples of eDNA are able to detect small crayfish populations, but not perfectly. Thus, we conclude that a program of repeated eDNA sampling is now feasible and likely reliable for measuring crayfish geographic range and for detecting new invasion fronts in the Honghe Hani landscape, which would inform regional control efforts and help to prevent the further spread of this invasive crayfish.

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Section: Introductionmentioning

confidence: 99%

Using eDNA to detect the distribution and density of invasive crayfish in the Honghe-Hani rice terrace World Heritage site

Cai

Yang

et al. 2017

PLoS ONE

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“…Consequently, the development of new technologies and tools allowing a hands‐off collection of DNA in the aquatic environment is suggested. Indeed, in the case of fishes, mucus (Livia et al , 2006) and scales (Yue & Orban, 2001) have already been reported to be suitable for DNA analyses, and, along with other sources of DNA ( e.g. faeces and urine), could be used in individual genetic tagging.…”

Section: Discussionmentioning

confidence: 99%

Individual genetic tagging for teleosts: an empirical validation and a guideline for ecologists

Andreou

Garcia

Blanchet

et al. 2011

Journal of Fish Biology

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The efficiency of individual genetic tagging was determined by using passive integrated transponders (PIT) as a comparative conventional tagging method. Fifty-five common dace Leuciscus leuciscus were captured in the wild, PIT tagged and fin clipped (for DNA analysis). Thirty fish were recaptured on three occasions and tissue samples were collected. Using 18 microsatellite loci, 79-94% of the recaptures were correctly assigned. Experience with scoring L. leuciscus microsatellites led to more individuals correctly assigned. Allowing matches that differed by one or two alleles resulted in 100% of all recaptures successfully assigned irrespective of the observer. Reducing the set of loci to five to six loci appropriately selected did not affect the assignment rate, demonstrating that costs can be subsequently reduced. Despite their potential benefits, the application of genetic tags for teleosts has been limited. Here, it was demonstrated that genetic tagging could be applied, and a clear guideline (flowchart) is provided on how this method can be developed for teleosts and other organisms, with subsequent practical applications to ecology, evolutionary biology and conservation management.

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“…It is now possible to extract enough DNA from many kinds of samples such as scales and body mucus, without harming the fish [13,14]. PCR-based techniques for sex identification using the sex-determination gene (SRY) or homologous fragments such as zinc finger protein genes (ZFX and ZFY) which are amplified together or separately have been used widely in many mammal species [15][16][17][18][19].…”

Section: Introductionmentioning

confidence: 99%

A PCR-based method for sex identification of critically endangered Formosa landlocked salmon

Hsu

Gwo

2010

Fish Sci

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The extinction risk in a small population depends largely on stochastic factors that affect effective population size. Unbalanced sex ratio, a form of demographic stochasticity, not only reduces the effective population size but also raises extinction risk. Sex identification is fundamental for conservation biology; however, it is extremely difficult to do in Formosa landlocked salmon Oncorhynchus masou formosanus. Because sexual dimorphism in landlocked dwarf form Formosa landlocked salmon is minimal, we developed a simple and noninvasive PCR-based method to identify sex in this critically endangered species. The OtY2m primers (derived from OtY2) amplified the male-specific marker (OtY2) in Formosa landlocked salmon. In addition, we provide a valuable tool for detection of sex ratios in wild populations for conservation management. This study contributes useful information to estimate the effective population size and analyze population viability of Formosa landlocked salmon.

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A nondestructive, rapid, reliable and inexpensive method to sample, store and extract high‐quality DNA from fish body mucus and buccal cells

Cited by 49 publications

References 14 publications

Using eDNA to detect the distribution and density of invasive crayfish in the Honghe-Hani rice terrace World Heritage site

Using eDNA to detect the distribution and density of invasive crayfish in the Honghe-Hani rice terrace World Heritage site

Individual genetic tagging for teleosts: an empirical validation and a guideline for ecologists

A PCR-based method for sex identification of critically endangered Formosa landlocked salmon

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