Abstract
In this study, 191 culture isolates were recovered from suspected samples of animals and humans in Ellinghausen McCullough Johnson and Harris (EMJH) medium and assessed for its morphological features by dark field microscopy. Extracted DNA from individual culture was subjected to different PCR assays for identification and characterization of leptospira. Out of 99 positive leptospira cultures, 52 pathogenic leptospira isolates were characterized at species level by using partial RNA polymerase β-subunit (rpoB) gene sequences. Phylogenetic analysis of the nucleotide sequences revealed that 30, 8, and 14 isolates belong to L. borgpetersenii / L. interrogans, L. kirschneri, and Leptospira intermediate species, respectively. Based on analysis of 99 leptospira isolates, the prevalent Leptospira species were L. borgpetersenii or L. interrogans (30.30%), L. kirschneri (8%) and Leptospira intermediate species (14.14%) in animals and humans. To the best of authors knowledge, this is the first study to use rpoB gene nucleotide sequence based phylogenetic analysis to identify/detect Leptospira intermediate species (L. wolffii) in animals and humans in India. Hence, the prevalence of this species will surely emphasize the importance of consideration of Leptospira intermediate species and formulate a way for further studies especially in understanding the newly emerging Leptospira in animals and humans and to combat the problem associated with the disease conditions.
Highlights
Leptospirosis is an important re-emerging zoonotic disease in tropical and subtropical regions of the world with various animal species acting as carriers
Clinical samples Clinical samples such as blood, serum, urine and other materials such as kidney, tissues and fluids from aborted fetus collected from suspected animals and humans [most of cases-pyrexia of unknown origin (PUO)] from different geographical locations of India collected either by the ADMAS team or obtained through various organizations such as state animal husbandry laboratories, research institutes and from organized farms which were submitted to the laboratory for leptospira diagnosis
Several molecular techniques have been evaluated for the identification and characterization of Leptospira spp. and DNA–DNA hybridization identified 20 Leptospira species to date with nine pathogenic Leptospira species (Cerqueira and Picardeau 2009)
Summary
Leptospirosis is an important re-emerging zoonotic disease in tropical and subtropical regions of the world with various animal species acting as carriers. The preliminary study was undertaken to investigate the prevalence of Leptospira species in animals and humans using the archived cultural isolates recovered from suspected samples of animals and humans in different geographical locality of India collected during different periods up to the species level by using rpoB gene- based sequence and phylogenetic analysis.
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