Pros and cons of external swabbing of amphibians for genetic analyses

Müller, Antonia S.; Lenhardt, Patrick P.; Theissinger, Kathrin

doi:10.1007/s10344-013-0747-2

Cited by 14 publications

(25 citation statements)

References 16 publications

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“…The selected loci were chosen from a number of tested loci due to their amplification success and polymorphism in an earlier study (Müller, Lenhardt & Theissinger, 2013). Also, six of the selected loci were located on different chromosomes (Table 1; see Cano et al, 2011).…”

Section: Methodsmentioning

confidence: 99%

Amphibian population genetics in agricultural landscapes: does viniculture drive the population structuring of the European common frog (Rana temporaria)?

Lenhardt

Brühl

Leeb

et al. 2017

PeerJ

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Amphibian populations have been declining globally over the past decades. The intensification of agriculture, habitat loss, fragmentation of populations and toxic substances in the environment are considered as driving factors for this decline. Today, about 50% of the area of Germany is used for agriculture and is inhabited by a diverse variety of 20 amphibian species. Of these, 19 are exhibiting declining populations. Due to the protection status of native amphibian species, it is important to evaluate the effect of land use and associated stressors (such as road mortality and pesticide toxicity) on the genetic population structure of amphibians in agricultural landscapes. We investigated the effects of viniculture on the genetic differentiation of European common frog (Rana temporaria) populations in Southern Palatinate (Germany). We analyzed microsatellite data of ten loci from ten breeding pond populations located within viniculture landscape and in the adjacent forest block and compared these results with a previously developed landscape permeability model. We tested for significant correlation of genetic population differentiation and landscape elements, including land use as well as roads and their associated traffic intensity, to explain the genetic structure in the study area. Genetic differentiation among forest populations was significantly lower (median pairwise FST = 0.0041 at 5.39 km to 0.0159 at 9.40 km distance) than between viniculture populations (median pairwise FST = 0.0215 at 2.34 km to 0.0987 at 2.39 km distance). Our analyses rejected isolation by distance based on roads and associated traffic intensity as the sole explanation of the genetic differentiation and suggest that the viniculture landscape has to be considered as a limiting barrier for R. temporaria migration, partially confirming the isolation of breeding ponds predicted by the landscape permeability model. Therefore, arable land may act as a sink habitat, inhibiting genetic exchange and causing genetic differentiation of pond populations in agricultural areas. In viniculture, pesticides could be a driving factor for the observed genetic impoverishment, since pesticides are more frequently applied than any other management measure and can be highly toxic for terrestrial life stages of amphibians.

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Section: Methodsmentioning

confidence: 99%

Amphibian population genetics in agricultural landscapes: does viniculture drive the population structuring of the European common frog (Rana temporaria)?

Lenhardt

Brühl

Leeb

et al. 2017

PeerJ

Self Cite

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“…Their characteristics do not only render them useful as a source of information on host genetics, they can also provide information on host health conditions, physiology and even interactions with micro‐organisms (Pidancier, Miquel & Miaud ; Handel et al . ; Allen & Torres ; Müller, Lenhardt & Theissinger ; Koboziev et al . ).…”

Section: Introductionmentioning

confidence: 99%

Protein and RNA extraction from mucosal swabs: a minimally invasive source of ecological data for studies of natural populations

et al. 2016

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Summary Mucosal swabs have long been used to study various physiological processes in humans. In contrast, systematic sampling of mucosae is rare for wild animals, and except for its use as a source of DNA for population genetics, its potential as a tool to study physiologically relevant processes of natural populations has not been explored fully. We collected swabs from the rectal, genital, nasal and oral mucosae of California sea lion pups and investigated differences in RNA and protein yield, quality and integrity between storage times and mucosal types. Downstream applications were tested on the extracted products to determine the potential value of mucosal sampling in free‐ranging mammals. For most samples, RNA yield was stable regardless of storage time, and RNA quality and integrity were equal for all mucosal types. Ribosomal fragments of sizes expected for mammals, yeast and bacteria were observable in genital and rectal samples, but in nasal and oral samples only bacterial ribosomal fragments were observable. Amplification of selected transcripts was successful for all samples. The protein profile was distinct between mucosae, and samples with high protein yields were useful for antibody detection. Our study demonstrates that mucosal swabbing is a minimally invasive tool that yields useful physiological data for free‐ranging wildlife. Minimum experience is needed to collect samples, processing is inexpensive, and downstream applications for ecological studies are realistic.

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“…While the operculum's success at blocking mucous sample contamination is not surprising, given its role in protecting sensitive gill filaments (Kimmel et al ., ), the authors note that care must be taken when collecting epidermal mucous samples from this location as they can easily be contaminated with blood from the gill filaments and may not necessarily be completely representative of epidermal mucus. In addition, because this sampling location is limited to bony fishes, the authors urge further investigation on the effects of epidermal mucous sample contamination based on variations in: morphology and physiology ( e.g ., body plan and fin structure, mucous production rate and mucous composition (Benhamed et al ., ; Shephard, ); behaviour and phenology [ e.g ., schooling or breeding (Müller et al ., )]; contact type, force and duration. Furthermore, we note that this work was conducted on fish carcasses that had been frozen and thawed, potentially influencing mucous composition and distribution.…”

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confidence: 99%

“…In addition, because this sampling location is limited to bony fishes, the authors urge further investigation on the effects of epidermal mucous sample contamination based on variations in: morphology and physiology (e.g., body plan and fin structure, mucous production rate and mucous composition (Benhamed et al, 2014;Shephard, 1994); behaviour and phenology [e.g., schooling or breeding (Müller et al, 2013)]; contact type, force and duration. Furthermore, we note that this work was conducted on fish carcasses that had been frozen and thawed, potentially influencing mucous composition and distribution.…”

mentioning

confidence: 99%

Exploring sample cross‐contamination in fish epidermal mucus

Kroska

Wolf

Dial

et al. 2019

Journal of Fish Biology

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Cross‐contamination of epidermal mucus was assessed at three sampling locations on the bodies of Pacific halibut Hippoglossus stenolepis by inducing contact between fish coated with labelled synthetic mucus and non‐treated fish. Results indicate a positive relationship between sampling site exposure and sample contamination and that mucous sample cross‐contamination can be mitigated by sampling in a location protected from external contact.

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Pros and cons of external swabbing of amphibians for genetic analyses

Cited by 14 publications

References 16 publications

Amphibian population genetics in agricultural landscapes: does viniculture drive the population structuring of the European common frog (Rana temporaria)?

Amphibian population genetics in agricultural landscapes: does viniculture drive the population structuring of the European common frog (Rana temporaria)?

Protein and RNA extraction from mucosal swabs: a minimally invasive source of ecological data for studies of natural populations

Exploring sample cross‐contamination in fish epidermal mucus

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