Sex identification of wolf (Canis lupus) using non-invasive samples

Sastre, Natalia; Francino, Olga; Lampreave, Gabriel; Bologov, Vladimir V.; López‐Martín, José María; Sánchez, Armand; Ramírez, Óscar

doi:10.1007/s10592-008-9565-6

Cited by 36 publications

(25 citation statements)

References 27 publications

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“…faeces, hair, etc.) have now been successfully applied to many studies of endangered mammals, such as population structure in brown bears (Perez et al 2009), sex determination in wolves (Sastre et al 2009), estimation of effective population sizes in giant pandas (Zhan et al 2006) and dietary habits in carnivores (Farrell et al 2000). This approach can be particularly useful in rare, endangered and secretive species like bongo.…”

Section: Introductionmentioning

confidence: 99%

The use of non-invasive molecular techniques to confirm the presence of mountain bongo Tragelaphus eurycerus isaaci populations in Kenya and preliminary inference of their mitochondrial genetic variation

et al. 2011

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The mountain bongo antelope Tragelaphus eurycerus isaaci has rapidly declined in recent decades, due to a combination of hunting, habitat degradation and disease. Endemic to Kenya, mountain bongo populations have shrunk to approximately 100 individuals now mainly confined to the Aberdares mountain ranges. Indirect observation of bongo signs (e.g. tracks, dung) can be misleading, thus methods to ensure reliable species identification, such as DNA-based techniques, are necessary to effectively study and monitor this species. We assessed bongo presence in four mountain habitats in Kenya (Mount Kenya National Park, Aberdare National Park, Eburu and Mau forests) and carried out a preliminary analysis of genetic variation by examining 466 bp of the first domain of the mtDNA control region using DNA extracted from faecal samples. Of the 201 dung samples collected in the field, 102 samples were molecularly identified as bongo, 97 as waterbuck, one as African buffalo and one as Aders' duiker. Overall species-identification accuracy by experienced trackers was 64%, with very high error of commission when identifying bongo sign (37%), and high error of omission for waterbuck sign (82%), suggesting that the two species' signs are easily confused. Despite high variation in the mtDNA control region in most antelope species, our results suggest low genetic variation in mountain bongo as only two haplotypes were detected in 102 samples analyzed. In contrast, the analysis of 63 waterbuck samples from the same sites revealed 21 haplotypes. Nevertheless, further examination using nuclear DNA markers (e.g. microsatellites) in a multi-locus approach is still required, especially because the use of mitochondrial DNA can result in population overestimation as distinct dung samples can potentially be originated from the same individual.

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Section: Introductionmentioning

confidence: 99%

The use of non-invasive molecular techniques to confirm the presence of mountain bongo Tragelaphus eurycerus isaaci populations in Kenya and preliminary inference of their mitochondrial genetic variation

et al. 2011

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“…DNA from tooth samples was amplified in a PCR mixture containing 30-40 ng of DNA, PCR buffer (19), 2.5 mM MgCl 2 , 0.2 mM of each dNTP, 0.4 lM of each primer and 1 U AmpliTaq Gold DNA Polymerase (Applied Biosystems). In order to avoid typing errors in tooth DNA sampleswith DNA of low quality and quantity-each sample was amplified four times (Sastre et al 2009). DNA from blood samples was amplified in two multiplex PCR reactions with three (MS34A, MS41A and 990) and two markers (MS34B, MS41B).…”

Section: Samplesmentioning

confidence: 99%

Signatures of demographic bottlenecks in European wolf populations

et al. 2010

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Monitoring the loss of genetic diversity in wild populations after a bottleneck event is a priority in conservation and management plans. Here, we used diverse molecular markers to search for signatures of demographic bottlenecks in two wolf populations; an isolated population from the Iberian Peninsula and a non-isolated population from European Russia. Autosomal, mtDNA and Y-chromosomal diversity and the effective population size (N e ) were significantly lower in the Iberian population. Neutrality tests using mtDNA sequences, such as R 2, Fu and Li's F*, Tajima's D and Fu's F s , were positively significant in the Iberian population, suggesting a population decline, but were not significant for the Russian population, likely due to its larger effective population size. However, three tests using autosomal data confirmed the occurrence of the genetic bottleneck in both populations. The M-ratio test was the only one providing significant results for both populations. Given the lack of consistency among the different tests, we recommend using multiple approaches to investigate possible past bottlenecks. The small effective population size (about 50) in the Iberian Peninsula compared to the presumed extant population size could indicate that the bottleneck was more powerful than initially suspected or an overestimation of the current population. The risks associated with small effective population sizes suggest that the genetic change in this population should be closely monitored in the future. On the other hand, the relatively small effective population size for Russian wolves (a few hundred individuals) could indicate some fragmentation, contrary to what is commonly assumed.

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“…However, detection and identification of species is difficult for rare and threatened taxa, mainly due to their low densities, nocturnal and elusive behaviour, and to the logistical and ethical difficulties involved in their capture and handling during traditional capture-markrecapture approaches. This has prompted the development of non-invasive tools applied to samples such as faeces, hair, feathers, urine, saliva, scent marks, sloughed skin and animal products illegally trafficked (e.g., Taberlet et al 1999;Dalèn et al 2004, Arrendal et al 2007Hogan et al 2008;Jones et al 2008;Sundqvist et al 2008;Sastre et al 2009). However, non-invasive samples seldom allow species identification based on morphology alone (Davison et al 2002).…”

Section: Introductionmentioning

confidence: 99%

Species identification using a small nuclear gene fragment: application to sympatric wild carnivores from South-western Europe

et al. 2009

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Species identification is essential for noninvasive studies of elusive and rare animals, and for detecting illegal harvest or trade of wildlife species. However, most molecular tests identify only a limited number of species or require multiple laboratory steps to distinguish many taxa. Additionally, most protocols use mitochondrial DNA being, therefore, especially prone to problems such as nuclear insert copies, high intraspecific diversity or heteroplasmy. Here, we developed a molecular test based on the polymorphisms detected on a small nuclear gene fragment (221 bp of the IRBP -Interphotoreceptor Retinoid-Binding Protein-exon 1). This fragment revealed 51 variable sites (including 12 non-synonymous and 19 species-specific sites), which enabled the successful distinction of all 16 carnivore species native to Southwestern Europe. A SSCP (Single-Strand Conformational Polymorphism) gel electrophoresis technique was also optimized to allow the simple and inexpensive application of this molecular test. Sequences and SSCP profiles were consistent in identifying a total of 387 samples, including faeces (172) and hairs (17) collected non-invasively in the field. Due to its low cost, simplicity, and wide range of identifiable species, this test shows great promise to facilitate studies in molecular ecology, conservation genetics, and forensic analysis, as well as DNA bar-coding projects.

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Sex identification of wolf (Canis lupus) using non-invasive samples

Cited by 36 publications

References 27 publications

The use of non-invasive molecular techniques to confirm the presence of mountain bongo Tragelaphus eurycerus isaaci populations in Kenya and preliminary inference of their mitochondrial genetic variation

The use of non-invasive molecular techniques to confirm the presence of mountain bongo Tragelaphus eurycerus isaaci populations in Kenya and preliminary inference of their mitochondrial genetic variation

Signatures of demographic bottlenecks in European wolf populations

Species identification using a small nuclear gene fragment: application to sympatric wild carnivores from South-western Europe

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