Identification of bat trypanosomes from Minas Gerais state, Brazil, based on 18S rDNA and Cathepsin-L-like targets

Abstract

Several bat species can be infected by trypanosomes, but there is not much information about which of these parasites infect bats from Triângulo Mineiro and Alto Paranaíba, Minas Gerais state, Brazil, a formerly endemic region for Trypanosoma cruzi, the causative agent of Chagas disease. The aim of this study was to describe, characterize, and identify the presence of trypanosomes in bats. The captured bats (448) belong to four families and to 19 different species. Of those, 37 bats were found to be positive for trypanosomes by microhematocrit, (infection rate 8.3%) and 27 were positive after hemoculture analysis. Initially, the isolates were identified by PCR (18S rDNA, 24Sα rDNA, spliced leader, COII RFLP-PCR) using primers originally designed for T. cruzi. PCRs (18S rDNA, 24Sα rDNA) showed compatible bands for TcI, whereas COII RFLP-PCR showed a similar pattern associated to TcII. However, there was no DNA amplification using spliced leader as a target, revealing a discrepancy between the results. Phylogenetic analysis of Cathepsin L-like and 18S rDNA sequences proved that 15 of the isolates corresponded to Trypanosoma cruzi marinkellei and one to Trypanosoma dionisii. These results revealed that the diversity of trypanosome species in a region considered endemic for Chagas disease is greater than previous descriptions. All this can confirm the necessity of using DNA sequencing approaches in order to determinate trypanosomes species isolated from bats.

Bat trypanosomatids (first report of T. wauwau) in Triângulo Mineiro, Brazil

“…Regarding PCR positivity, the prevalence of trypanosomatids was similar to that observed by other studies in Brazil [3][4], as was the result that in terms of dietary habits most of these bats were nectarivores. It was also demonstrated in this study that the two most common species that circulate in bats in this region are T. c. marinkellei and T.…”

“…In this study, the use of the direct parasitological method of microhematocrit revealed frequent positive results for trypanosomatids, as has been demonstrated by other studies [4,28]. It is worth highlighting that the microhematocrit method, in all the field studies carried out by our group, especially in small animals, has shown greater sensitivity or, in certain cases, similar behavior to that of traditional blood culture methods [29].…”

“…Some species of trypanosomatids have already been found to naturally infect bats [4,[7][8][9][10][24][25][26][27]. Many of these trypanosomatids are responsible for important zoonoses, but most of those found in bats remain poorly known.…”

“…Bats were captured from November 2014 to September 2015, at night between 18h00 and midnight (0h00) using mist nets, and during the day in shelters using manual 4 nets. The identification of bats species was done based on taxonomic keys to the family [17], genus, and species levels [18][19].…”

“…Until now, more than up to 30 species of trypanosomatids have been isolated from bats [1][2][3][4]. The most frequently reported species of Trypanosoma in bats are T. cruzi, T. cruzi marinkellei, T. dionisii, T. hedricki, T. myoti, T. leonidasdeanei, T. desterrensis, T. pifanoi, T. pessoai, T. megaderma-like, T. theileri, and T. rangeli [5][6][7][8][9][10].…”