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Research article - Peer-reviewed, 2015

A New Method for Noninvasive Genetic Sampling of Saliva in Ecological Research

Lobo D, Godinho R, Alvares F, Lopez-Bao JV, Rodriguez A

Abstract

Noninvasive samples for genetic analyses have become essential to address ecological questions. Popular noninvasive samples such as faeces contain degraded DNA which may compromise genotyping success. Saliva is an excellent alternative DNA source but scarcity of suitable collection methods makes its use anecdotal in field ecological studies. We develop a noninvasive method of collection that combines baits and porous materials able to capture saliva. We report its potential in optimal conditions, using confined dogs and collecting saliva early after deposition. DNA concentration in saliva extracts was generally high (mean 14 ng mu l(-1)). We correctly identified individuals in 78% of samples conservatively using ten microsatellite loci, and 90% of samples using only eight loci. Consensus genotypes closely matched reference genotypes obtained from hair DNA (99% of identification successes and 91% of failures). Mean genotyping effort needed for identification using ten loci was 2.2 replicates. Genotyping errors occurred at a very low frequency (allelic dropout: 2.3%; false alleles: 1.5%). Individual identification success increased with duration of substrate handling inside dog's mouth and the volume of saliva collected. Low identification success was associated with baits rich in DNA-oxidant polyphenols and DNA concentrations <1 ng mu l(-1). The procedure performed at least as well as other noninvasive methods, and could advantageously allow detection of socially low-ranked individuals underrepresented in sources of DNA that are involved in marking behaviour (faeces or urine). Once adapted and refined, there is promise for this technique to allow potentially high rates of individual identification in ecological field studies requiring noninvasive sampling of wild vertebrates.

Published in

PLoS ONE
2015, volume: 10, number: 10, article number: e0139765
Publisher: Public Library of Science

Authors' information

Lobo, Diana
University of Porto
Godinho, Raquel
University of Porto
Álvares, Francisco
University of Porto
López-Bao, José Vicente
Swedish University of Agricultural Sciences, Department of Ecology
López-Bao, José Vicente
University of Oviedo
Rodríguez, Alejandro
Consejo Superior de Investigaciones Cientificas (CSIC)

UKÄ Subject classification

Genetics
Ecology

Publication Identifiers

DOI: https://doi.org/10.1371/journal.pone.0139765

URI (permanent link to this page)

https://res.slu.se/id/publ/70179