Pathogenic Leptospira Species in Bats: Molecular Detection in a Colombian Cave

Abstract

Leptospirosis is caused by pathogenic Leptospira spp., which can be found in nature among domestic and wild animals. In Colombia, the Macaregua cave is known for its bat richness; thus, because bats are reservoir hosts of human microbiological pathogens, we determined if the Macaregua cave bats harbored Leptospira in the wild. A total of 85 kidney samples were collected from three bat species (Carollia perspicillata, Mormoops megalophylla, and Natalus tumidirostris) to detect Leptospira spp. The 16S rRNA gene was targeted through conventional PCR and qPCR; in addition, the LipL32 gene was detected using conventional PCR. Obtained amplicons were purified and sequenced for phylogenetic analysis. The Leptospira spp. 16S rRNA gene was detected in 51.8% bat kidneys, of which 35 sequences were obtained, all clustering within the pathogenic group. Moreover, 11 sequences presented high-identity-values with Leptospiranoguchii, Leptospiraalexanderi, Leptospiraborgpetersenii, Leptospirakirschneri, and Leptospiramayottensis. From the 16S rRNALeptospira spp.-positive population samples, 28 amplified for the LipL32 gene, and 23 sequences clustered in five different phylogenetic groups. In conclusion, we detected the circulation of different groups of Leptospira spp. sequences among cave bats in the wild; some sequences were detected in more than one bat specimen from the same species, suggesting a conspecific transmission within the cave.

Keywords: Colombia; Leptospira; bats; leptospirosis.

Figure 1

Leptospira spp. 16S rRNA sequence-based phylogenetic tree detected in bats. Sequences retrieved from this study are indicated by symbols: black circles from N. tumidirostris, black squares from M. megalophylla, and black rhombuses from C. perspicillata. The GenBank numbers from the reference sequences are indicated in brackets, and the Leptospira spp. sequences obtained from previous Colombian bat species studies are indicated by white rhombuses [21]. The Leptospira species and their groups are listed to the right of each branch.

Figure 2

LipL32 gene sequence-based phylogenetic tree for Leptospira spp. detected in bats. The sequences retrieved in this study are indicated by symbols: blue circles from M. megalophylla and red squares from N. tumidirostris; numbers in brackets in MZ787849, MZ787850, MZ787851, and MZ787848 sequences represent the number of sequences obtained from bat specimens; sequences without a particular number in brackets depict that only a sequence was obtained. GenBank numbers from reference sequences are indicated in brackets, and Leptospira spp. sequences obtained from bat species from previous studies are indicated by black rhombuses and black triangles [21,31]. Leptospira groups and identity percentages between sequences for each group are illustrated.