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Optimization of protocols for DNA extraction from fecal samples

High-throughput sequencing offers new possibilities in molecular ecology and conservation studies. However, its potential has not yet become fully exploited for noninvasive studies of free–ranging animals, such as those based on feces. High–throughput sequencing allows sequencing of short DNA fragments and could allow simultaneous genotyping of a very large number of samples and markers at a low cost. The application of high throughput genotyping to fecal samples from wildlife has been hindered by several labor intensive steps. Alternative protocols which could allow higher throughput were evaluated for two of these steps: sample collection and DNA extraction. Two different field sampling and seven different DNA extraction methods were tested on grey wolf (Canis lupus) feces. There was high variation in genotyping success rates. The field sampling method based on surface swabbing performed much worse than the extraction from a fecal fragment. In addition, there is a lot of room for improvement in the DNA extraction step. Optimization of protocols can lead to very much more efficient, cheaper and higher throughput noninvasive monitoring. Selection of appropriate markers is still of paramount importance to increase genotyping success. informacion[at] Sarabia et al (2020) Towards high-throughput analyses of fecal samples from wildlife. Animal Biodiver Conserv 43.2: 271–283 Doi 10.32800/abc.2020.43.0271
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Vertebrate road-kill patterns in Mediterranean habitats: who, when and where.

Vertebrate road-kill patterns in Mediterranean habitats: who, when and where.

Road-kill is the most recognized impact of traffic and an important threat for biodiversity. Nevertheless, most research on this topic deals with particular species or with road features, describing proximate correlates and rarely making inference on the mechanisms. Here it is provided a more general approximation by describing life-history, temporal and spatial factors affecting vertebrate road-kills in Mediterranean landscapes, a biodiversity hotspot with little studied road impacts. During one year authors recorded the casualties found on paved roads within Doñana Natural Park. More than 2300 road-kills belonging to 66 species (32% of the study area checklist) were found, with abundant ectotherm species more likely to be road-killed. The temporal and spatial factors affecting the road-kill patterns of different taxonomic and functional groups were also investigated. The phenology of the species was the main factor affecting road-kill temporal patterns for lizards, all birds and small mammals. Additionally, rainfall events were associated with the road-kill peaks of wintering birds, whereas high temperatures were related to the increase of road-killed snakes and the decrease of road-killed amphibians. Amphibians, snakes, lizards and small passerines were mainly road-killed according with their spatial abundance. Mitigation measures such as wildlife road-crossing structures showed contradictory effectiveness for small vertebrates due to the lack of adequate drift fences. Authors suggest prioritizing the mitigation measures which can permanently decrease the risk of been road-killed for ectotherm species, such as specific road-crossing structures with effective drift fences on road-kill hotspots. Concurrently, group-specific temporal mitigation measures should be applied during the road-kill seasonal peaks. The present work provides recommendations to decrease road-kill impacts in Mediterranean environments, but simultaneously tries to contribute to a more general development of road ecology research, suggesting several useful guidelines to perform road-kill studies. informacion[at] D'Amico et al (2015) Vertebrate road-kill patterns in Mediterranean habitats: Who, when and where. Biol Conserv 191: 234–242 DOI: 10.1016/j.biocon.2015.06.010