The bands were visualised using a UV transilluminator learn more after ethidium bromide staining (0.5 μg/mL). The amplicons were purified using the QIAquick® PCR and the QIAEX II kits (Qiagen) for the H. capsulatum and Pneumocystis organisms, respectively. Afterwards, the amplicons were sent to the Molecular Biology Laboratory, Institute of Cellular Physiology (UNAM, Mexico) for sequencing in an ABI-automated DNA sequencer (Applied Biosystems Inc., Foster City, CA, USA). Sequencing reactions were performed for forward and reverse

DNA strands, and a consensus sequence for each amplified bat lung sample product was generated. The sequences were edited and aligned using the MEGA software, version 5 (http://​www.​megasoftware.​net). Most of the Hcp100 selleck sequences of H. capsulatum were previously reported in González-González et al. [6], and the other sequences were deposited in a database [GenBank: from JX091346 to JX091370 accession numbers]. All sequences

generated by both molecular markers for Pneumocystis spp. were reported by Derouiche et al. [16] and Akbar et al. [14]. The sequences of the specific markers for each pathogen (i.e., Hcp100 for H. capsulatum and mtLSUrRNA or mtSSUrRNA for Pneumocystis spp.) that were obtained in the same animal were the main inclusion criterion for considering bat co-infection. Statistics The infection and co-infection rates for each pathogen were estimated by considering all of the bats studied from the three countries and from each country separately (Argentina, French Guyana, and Mexico), in relation to those bats with H. capsulatum and Pneumocystis spp. infections as identified by sequencing their respective molecular markers. The corresponding 95% confidence interval (CI) was calculated using a normal

distribution. Results Data from nine bat species studied belonging to five different families, highlighting their particular behaviours, such as migration, nourishment, distribution in the American continent and colony size, are referred to in Table 1, according to Ceballos and Oliva Nitroxoline [23]. These behaviours varied considerably among the bat species studied (Table 1). The different species captured, their numbers, and their geographical origins are registered in Table 2. Although most of the bat species studied were non-migratory, the number of https://www.selleckchem.com/products/cilengitide-emd-121974-nsc-707544.html migratory bats from three processed species was greater than that of the non-migratory species (Tables 1 and 2). It is noteworthy that among the 122 bats studied, 84 (68.80%) belonged to the migratory species Tadarida brasiliensis, from which 63 individuals were captured in Mexico and 21 in Argentina (Table 2).