Barros Juliana Helena da Silva, Roque André Luiz Rodrigues, Xavier Samanta Cristina das Chagas, Nascimento Kátia Cristina Silva, Toma Helena Keiko, Madeira Maria de Fatima
Laboratório de Biologia de Tripanosomatídeos, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil.
Laboratório de Diagnóstico Molecular e Hematologia, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21491-599, Brazil.
Pathogens. 2020 Sep 7;9(9):736. doi: 10.3390/pathogens9090736.
This study describes the morphological, biochemical, and molecular differences among isolates from hemocultures of hematophagous (; = 2) and insectivorous (; = 1) bats from the Atlantic Rainforest of Rio de Janeiro, Brazil. Fusiform epimastigotes from the hematophagous isolates were elongated, whereas those of the insectivorous isolate were stumpy, reflected in statistically evident differences in the cell body and flagellum lengths. In the hemocultures, a higher percentage of trypomastigote forms (60%) was observed in the hematophagous bat isolates than that in the isolate from the insectivorous bat (4%), which demonstrated globular morphology. Three molecular DNA regions were analyzed: V7V8 (18S rDNA), glycosomal glyceraldehyde 3-phosphate dehydrogenase gene, and mitochondrial cytochrome gene. The samples were also subjected to multilocus enzyme electrophoresis and random amplified polymorphic DNA analysis. All isolates were identified as by phylogenetic analysis. These sequences were clustered into two separate subgroups with high bootstrap values according to the feeding habits of the bats from which the parasites were isolated. However, other samples from bats with different feeding habits were found in the same branch. These results support the separation of the three isolates into two subgroups, demonstrating that different subpopulations of circulate among bats.
本研究描述了来自巴西里约热内卢大西洋雨林的吸血蝙蝠(n = 2)和食虫蝙蝠(n = 1)血培养物分离株之间的形态、生化和分子差异。吸血蝙蝠分离株的梭形上鞭毛体细长,而食虫蝙蝠分离株的则粗短,这反映在细胞体和鞭毛长度上有统计学上明显的差异。在血培养物中,吸血蝙蝠分离株中观察到的锥鞭毛体形式百分比(60%)高于食虫蝙蝠分离株(4%),后者表现为球形形态。分析了三个分子DNA区域:V7V8(18S rDNA)、糖体甘油醛-3-磷酸脱氢酶基因和线粒体细胞色素基因。样本还进行了多位点酶电泳和随机扩增多态性DNA分析。通过系统发育分析,所有分离株均被鉴定为克氏锥虫。根据分离出寄生虫的蝙蝠的摄食习性,这些序列以高自展值聚为两个单独的亚组。然而,在同一分支中发现了来自具有不同摄食习性蝙蝠的其他克氏锥虫样本。这些结果支持将这三个分离株分为两个亚组,表明不同亚群的克氏锥虫在蝙蝠之间传播。