Molecular detection and characterization of Leptospira species in bats and other small wild mammals from Villeta municipality, Colombia

Abstract

Leptospira is a bacterial genus which includes several pathogenic species. Wild mammals can act as reservoir hosts, shedding bacteria in their urine. Leptospirosis is an important health problem in Villeta, but data regarding potential reservoirs hosts and natural sources of infection are still scarce. We aimed to detect and characterize the presence of Leptospira among small wild mammals from Villeta municipality, Colombia. Small wild mammals from three orders: Didelphimorphia, Chiroptera and Rodentia, were sampled in the region. DNA was extracted from kidney samples and screened for Leptospira through real-time PCR targeting a the 16 s rRNA gene. Positive samples were screened through conventional PCR using five complementary genes: adk, icdA, lipL32, lipL41 and secY. Amplicons were sequenced and used for phylogenetic analysis. A total of 75 kidney samples were collected from three small wild mammal orders: 62 bats, 5 opossums, and 7 rodents. Leptospira spp. was detected in 38.7 % of the samples, with bats presenting the most frequent infection rate (43.5 %). The infection rate varied by sampling site, with the highest frequency observed in Mave village (57.1 %). Concatenated phylogenetic analysis revealed that all sequences clustered within the P1/Pathogenic Leptospira major clade, forming three subclades: two bat-related Leptospira subclades, and one subclade including Leptospira interrogans, Leptospira kirschneri, and Leptospira noguchii species. This study describes the presence of Leptospira among bats and opossums from Villeta, Colombia. It also identifies the circulation of several P1/pathogenic Leptospira species among bats forming three clusters, two of them composed exclusively of bat-related leptospires, and one together with recognized pathogenic species.