Rapid assessment of heavy metal toxicity using bioluminescent bacteria Photobacterium leiognathi strain GoMGm1.

Several commercial test kits such as Microtox, LUMIStox, ToxAlert, Aboatox, and ToxScreen have been widely used for toxicity screening. Though this time saving assays offer excellent sensitivity, cost-effectiveness, and accuracy, these commercial assays are limited in terms of real-time monitoring in Indian coastal environment due to warmer temperatures. This necessitates the need to develop a rapid and accurate assay that can be effectively employed for real time monitoring with respect to heavy metals in the Indian coastal waters. With this objective, the present study was conducted by isolating an indigenous luminescent bacterium from the light organs of chordates Gazza minuta which showed higher luminescence in a wide range of temperatures. The isolate could grow well in the temperature of 30 ± 2 °C and withstand temperature up to 35 ± 2 °C. The isolated bacterium was identified as Photobacterium leiognathi GoMGm1 based on 16S rDNA and luxA gene sequences. The suitable growing medium was optimized using central composite rotational design (CCRD) method to obtain optimal growth and luminescence. The optimized medium exemplified the maximal growth and luminescence of P. leiognathi at OD600 nm of 5.78 ± 0.12 and RLU of 12.49 ± 0.43. The isolate was used to assess the toxicity of several heavy metals. The IC50 values of 0.0051, 1.13, 1.37, 3.1, and 6.68 mg L-1 were observed for the Hg, Cr, Cu, Ni, and Zn, respectively, after 15 min of exposure. Results obtained from principal component analysis (PCA) displayed the present assay's compatibility with other luminescent bacterial assay and commercial Microtox™ assay. Thus, it would the right candidate as an early detection system for heavy metals in aquatic bodies in tropical countries. Schematic representation of the present study.

New Delhi Metallo-ß-lactamase and other mechanisms of carbapenemases among Enterobacteriaceae in rural South India.

OBJECTIVES: The emergence and dissemination of carbapenem-resistant Enterobacteriaceae (CRE) is an important public health problem. This study aimed to understand the prevalence and mechanisms of carbapenem resistance in clinically important members of Enterobacteriaceae in rural South India. METHODS: Routine clinical isolates of Escherichia coli and Klebsiella spp. were tested for Ertapenem (ETP) non-susceptibility by the disk diffusion method over a 3-year period (2012-2014). The ETP non-susceptible isolates were preserved, and tested for the MIC of carbapenems and the carriage of major carbapenemase-encoding genes. Representative genes were sequenced and selective isolates were tested for the production of carbapenemase by carbapenem inactivation method. RESULTS: A total of 444 ETP non-susceptible isolates were identified in increasing incidence over the study period. Among them, MIC50 and MIC90 of carbapenems (excluding ETP) were 0.25-0.5µg/mL and 8-16µg/mL, respectively, and the prevalence of non-ETP carbapenem resistance was estimated as 3%. Among the 177 tested isolates, 65 (37%) had one or more carbapenemase-encoding genes with a predominance of New Delhi Metallo-ß-lactamase (NDM; 32 of 65; 49.2%). CONCLUSIONS: This study documented the MIC range for carbapenems, prevalence and mechanisms of carbapenem resistance among Enterobacteriaceae in rural South India. It substantiated NDM as a leading mechanism of carbapenem resistance and highlighted the importance of MIC testing in patient management.

In silico prediction of potential vaccine candidates on capsid protein of human bocavirus 1.

Human bocavirus 1 (HBoV1) is a newly identified parvovirus that causes serious respiratory infection among children across the globe. Aim of the present study was to predict immunogenic residues located on the VP2 protein of HBoV1 towards development of epitope based vaccines. Several computational tools were employed to predict epitopes (bothT and B cell restricted) with stringent regulation for the improvement of confidence. After meticulous analysis, the peptide "TTPWTYFNFNQY" was identified as potential candidate for development of preventive vaccine. Of note, the epitope "TTPWTYFNFNQY" was found to be recognized by fifteen different alleles belonging to seven HLA supertypes (A1, A3, A24, A26, B7, B58 and B62). Further, mutational variability analysis pointed that most of the amino acids were well conserved. Docking scores obtained from ClusPro and Autodock Vina for selected epitopes displayed energetically favorable and stable interaction of peptide-HLA-I complexes. The core peptide "LLYQMPFFL" was found to recognize by wide range of HLA class II allele recognition thereby qualified as candidate for therapeutic vaccine. Five distinct linear peptides (withT cell epitope superimposition) belonging to B cells were identified in the VP2 protein. Further attention on the enlisted epitopes may shed light on the path for development of diagnostic, therapeutic and preventive tools against HBoV1 infection. Additionally, the predicted epitopes may help us to address the original antigenic sin phenomena observed during consecutive HBoV2-4 infection.

Gut microbial degradation of organophosphate insecticides-induces glucose intolerance via gluconeogenesis.

BACKGROUND: Organophosphates are the most frequently and largely applied insecticide in the world due to their biodegradable nature. Gut microbes were shown to degrade organophosphates and cause intestinal dysfunction. The diabetogenic nature of organophosphates was recently reported but the underlying molecular mechanism is unclear. We aimed to understand the role of gut microbiota in organophosphate-induced hyperglycemia and to unravel the molecular mechanism behind this process. RESULTS: Here we demonstrate a high prevalence of diabetes among people directly exposed to organophosphates in rural India (n = 3080). Correlation and linear regression analysis reveal a strong association between plasma organophosphate residues and HbA1c but no association with acetylcholine esterase was noticed. Chronic treatment of mice with organophosphate for 180 days confirms the induction of glucose intolerance with no significant change in acetylcholine esterase. Further fecal transplantation and culture transplantation experiments confirm the involvement of gut microbiota in organophosphate-induced glucose intolerance. Intestinal metatranscriptomic and host metabolomic analyses reveal that gut microbial organophosphate degradation produces short chain fatty acids like acetic acid, which induces gluconeogenesis and thereby accounts for glucose intolerance. Plasma organophosphate residues are positively correlated with fecal esterase activity and acetate level of human diabetes. CONCLUSION: Collectively, our results implicate gluconeogenesis as the key mechanism behind organophosphate-induced hyperglycemia, mediated by the organophosphate-degrading potential of gut microbiota. This study reveals the gut microbiome-mediated diabetogenic nature of organophosphates and hence that the usage of these insecticides should be reconsidered.

KIR : HLA association with clinical manifestations of HBV infection in Madurai, south India.

The antiviral action of natural killer (NK) cells is regulated by a wide repertoire of germ-line encoded membrane receptors which recognize the expression of certain self-molecules on target cells. Among the receptors, killer cell immunoglobulinlike receptor (KIR) which recognizes the expression of human leukocyte antigen (HLA) class I has a predominant role in regulating the effector functions of NK cells, particularly in viral infections.We studied a total of 128 hepatitis B virus (HBV) patients (15 acute, 43 asymptomatic, 27 chronic and 43 with other liver diseases) while attending the Department of Medical Gastroenterology, Government Rajaji Hospital, Madurai, India, and 128 ethnic matched control to find the association between the KIR : HLA genes and differential manifestations of HBV. KIR and its ligand HLA polymorphism were identified by DNAPCR methods. The activatory receptor KIR-2DS1 was significantly elevated in various disease categories, namely asymptomatic, chronic and other HBV, except acute HBV infection. Whereas, KIR 2DS3 in acute and chronic patients and KIR 2DS5 and 3DS1 in asymptomatic individuals. Among various KIR-HLA combinations, homozygous 2DS2:C1 and individuals with 3DSI:BW4 (OR = 3.23, CI = 1.55-6.7, Pc = 0.02) are associated with HBV asymptomatism, while most of the two domain inhibitory receptors with their ligands showed significant risk in other liver diseases. Further, KIR3DL1 : HLA Bw4Iso80 (OR = 3.89, 95% CI = 1.58-9.55, Pc = 0.004) is related with higher risk for asymptomatic infection when compared with chronic HBV. Thus, the select KIR : HLA alleles and combinations seem to direct the NK cell activities and immune response in different directions resulting in varied symptoms and manifestations in the subgroups of HBV-infected patients studied.