Using DNA Barcodes to Identify Road-Killed Animals in Two Atlantic Forest Nature Reserves, Brazil

Abstract

Road mortality is the leading source of biodiversity loss in the world, especially due to fragmentation of natural habitats and loss of wildlife. The survey of the main species victims of roadkill is of fundamental importance for the better understanding of the problem, being necessary, for this, the correct species identification. The aim of this study was to verify if DNA barcodes can be applied to identify road-killed samples that often cannot be determined morphologically. For this purpose, 222 vertebrate samples were collected in a stretch of the BR-101 highway that crosses two Discovery Coast Atlantic Forest Natural Reserves, the Sooretama Biological Reserve and the Vale Natural Reserve, in Espírito Santo, Brazil. The mitochondrial COI gene was amplified, sequenced and confronted with the BOLD database. It was possible to identify 62.16% of samples, totaling 62 different species, including Pyrrhura cruentata, Chaetomys subspinosus, Puma yagouaroundi and Leopardus wiedii considered Vulnerable in the National Official List of Species of Endangered Wildlife. The most commonly identified animals were a bat (Molossus molossus), an opossum (Didelphis aurita) and a frog (Trachycephalus mesophaeus) species. Only one reptile was identified using the technique, probably due to lack of reference sequences in BOLD. These data may contribute to a better understanding of the impact of roads on species biodiversity loss and to introduce the DNA barcode technique to road ecology scenarios.

Using DNA barcodes to identify forensically important species of Diptera in Espírito Santo State, Brazil

“…Regarding the use of universal primers, matches of >99% of similarity were retrieved for only 10/28 (35.7%) samples using BOLD database (Figure 2). Low levels of identification using BOLD were also described by Klippel et al (2015) when studying roadkilled animals. Using COI gene it was possible to identify 62.16% of samples, however, the percentage varied from about 87.5% when studying non-volant mammals to 0% in reptiles.…”

Roadkill records of reptiles and birds in Cerrado and Pantanal landscapes

“…First geographic records (FGR) and new roadkill records (NRR) for reptile and bird species were verified from the literature. We compared our dataset to data collected by De Souza et al (2014) from the same highway (BR-262) and data of road-killed reptiles and birds from other Brazilian roads (Novelli et al 1988, Prado et al 2006, Silva et al 2007, Coelho et al 2008, Hengemühle and Cademartori 2008, Cunha et al 2010, Hartmann et al 2011, Ramos et al 2011, Santana 2012, Santos et al 2012, 2014Carvalho et al 2014, Klippel et al 2015, Machado et al 2015, Almeida et al 2016, Braz and França 2016, Deffaci et al 2016, Saranholi et al 2016, Steil et al 2016, Corrêa et al 2017, Miranda et al 2017, Rodríguez-Castro et al 2017, Viturino and Graciolli 2017, Gonçalves et al 2018, Ramos-Abrantes et al 2018.…”

Highways are a threat for giant armadillos that underpasses can mitigate

“…They were located at a 5.1 km section, between km 102 and km 107 of the BR‐101 highway, which intercepts the SBR. This highway divides the SBR conservation unit into two fragments, east and west (Klippel et al, 2015). The structures were rainwater drainage culverts and had three different shapes: circular, rectangular and a shape as a tunnel, ranging 0.9 m x 1.1 m, 1.1 m × 1.3 m and 1.7 m x 1.1 m (height × width), respectively.…”

“…While there are no vehicle collision with giant armadillos recorded in literature on this highway, other animals were frequently recorded (Gumier‐Costa & Sperber, 2009). In the Atlantic Forest, there is currently no record for roadkill of giant armadillos, although many other species are victims of vehicle collisions in the SBR and VNR (Klippel et al, 2015), including large mammals (Srbek‐Araujo, Mendes, & Chiarello, 2015). Overall, only a small fraction of roads crossing the distribution of giant armadillos in Brazil were monitored and for this reason it is safe to assume that many more animals were killed by vehicle collisions during this period.…”