BackgroundLeptospirosis is a re-emerging infectious disease that is under-recognized due to low-sensitivity and cumbersome serological tests. MAT is the gold standard test and it is the only serogroup specific test used till date. Rapid reliable alternative serogroup specific tests are needed for surveillance studies to identify locally circulating serogroups in the study area.Methods/Principal FindingsIn the present investigation the serological specificity of leptospiral lipopolysaccharides (LPS) was evaluated by enzyme linked immunosorbent assay (ELISA), dot blot assay and rapid immunochromatography based lateral flow assay (ICG-LFA). Sera samples from 120 MAT positive cases, 174 cases with febrile illness other than leptospirosis, and 121 seronegative healthy controls were evaluated for the diagnostic sensitivity and specificity of the developed assays. LPS was extracted from five locally predominant circulating serogroups including: Australis (27.5%), Autumnalis (11.7%), Ballum (25.8%), Grippotyphosa (12.5%), Pomona (10%) and were used as antigens in the diagnostics to detect IgM antibodies in patients’ sera. The sensitivity observed by IgM ELISA and dot blot assay using various leptospiral LPS was >90% for homologous sera. Except for Ballum LPS, no other LPS showed cross-reactivity to heterologous sera. An attempt was made to develop LPS based ICG-LFA for rapid and sensitive serogroup specific diagnostics of leptospirosis. The developed ICG-LFA showed sensitivity in the range between 93 and 100% for homologous sera. The Wilcoxon analysis showed LPS based ICG-LFA did not differ significantly from the gold standard MAT (P>0.05).ConclusionThe application of single array of LPS for serogroup specific diagnosis is first of its kind. The developed assay could potentially be evaluated and employed for as MAT alternative.

Leptospirosis is mainly considered an occupational disease, prevalent among agriculture, sewage works, forestry, and animal slaughtering populations. However, putative risk to miners and their inclusion in the high-risk leptospirosis group remain in need of rigorous analysis. Therefore, a study was conducted with the objective to assess the leptospirosis seroprevalence among miners of two districts of Tamil Nadu, India. A total of 244 sera samples from Pudukkottai miners (124) and Karur miners (120) were analyzed by microscopic agglutination test. Antibodies to leptospires were detected in 94 samples giving an overall seroprevalence of 38.5%. The seroprevalence was higher among Pudukkottai miners (65.3%) when compared with Karur miners (10.8%). Seroprevalence among control population (13%) was significantly less than that of the Pudukkottai miners marking a possible high-risk population group distinction. Subject sera most commonly reacted with organisms of the serogroup Autumnalis, and the pattern was similar in carrier animals of the study areas. Two leptospires were isolated from kidney samples of rats. The prevalence of Autumnalis among rodents and humans source tracked human leptospirosis among the miners. The study also determined that Pudukkottai miners are subjected to high-risk challenges such as exposure to water bodies on the way to the mines (odds ratio [OR] = 10.6), wet mine areas (OR = 10.6), rat infestation (OR = 4.6), and cattle rearing (OR = 10.4) and are thus frequently exposed to leptospirosis compared with Karur miners. Hence, control strategies targeting these populations will likely to prove to be effective remediation strategies benefiting Pudukkottai miners and workers in similar environments across occupations.

Humans are known to excrete leptospires in their urine after recovery from illness and there are reports showing development of asymptomatic leptospiruria in settings of high disease transmission. In this regard, we sought to evaluate the asymptomatic renal carriage status of humans in the highly endemic region of Tiruchirapalli district, Tamilnadu, India. A total of 245 asymptomatic participants were included. Urine and blood samples were collected and the extent of leptospiral infection was characterized by MAT, qPCR, 16S rRNA, and dot blot assay. The qPCR screening with urine DNA identified 129 (52.7%) positive samples further confirmed by nested PCR. The dot blot assay marked 30.2% (74/245) as true positives. The phylogenetic analysis showed the sequences to cluster with pathogenic Leptospira spp. Serological results showed 50 people with urine positivity to be negative for MAT and can probably be classified as 'asymptomatic individuals.' In conclusion, it can be speculated that in endemic regions there is a greater possibility of humans as maintenance host rather than incidental hosts.

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Leptospirosis is a worldwide reemerging tropical zoonotic disease with symptoms of mild febrile illness to more severe multiple organ failure caused by pathogenic leptospiral strains. There was no effective antibiotic for treating leptospirosis. Here, the anti-leptospiral potential of marine actinobacterial compound from Streptomyces indiaensis MSU5 isolated from Manakudy marine sediment, Tamil Nadu, India was evaluated. The potential actinobacterial strain was identified by phenotypic, cell wall, 16S rRNA gene sequence and phylogenetic analysis. In vitro anti-leptospiral activity of the actinobacterial compound was determined using broth microdilution test against various serovars of Leptospira with different concentration ranging from 15.625 to 500 µg/ml. Mass production of anti-leptospiral compound was carried out in agar surface fermentation with optimized condition and purified by preparative TLC. The purified fraction of anti-leptospiral compound named as MSU5-1, and it was confirmed by microdilution test. Remarkably, the compound MSU5-1 showed minimum inhibitory concentration of 62.5 µg/ml and minimum bactericidal concentration of 125 µg/ml against human pathogenic leptospiral isolate strain N2. The structural elucidation of purified compound was carried out using UV, FT-IR, NMR and LC-MS analysis. The compound MSU5-1 was tentatively identified as leptomycin B (CHO) with molecular weight 541.1 g/mol. Anti-leptospiral activity of compound MSU5-1 exhibited 80% of survival rate in mice model, further it was confirmed by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) analysis. From the available literature, this is the first report on the marine actinobacterial compound for evaluating both in vitro and in vivo leptospiricidal activity.