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Dolker S.,Indian Veterinary Research Institute | Verma R.,Indian Veterinary Research Institute | Prakash C.,Indian Veterinary Research Institute | Shamal A.,Indian Veterinary Research Institute | Shamal A.,Center for Animal Disease Research and Diagnosis
Indian Journal of Animal Sciences | Year: 2012

Diagnostic potentiality of loop mediated isothermal amplification (LAMP) assay with human sputum samples was compared with smear microscopy, culture and PCR in providing rapid and accurate TB diagnosis. Practical application of this assay was emphasized as a supplementary diagnostic test with acid fast smear microscopy for controlling tuberculosis in limited resource setting particularly India. Sputum samples (261) from TB suspected patients were collected, processed and subjected to smear microscopy, culture, PCR and LAMP assay. These 4 diagnostic methodologies were compared in terms of sensitivity, specificity, and rapidity and user friendliness. Sensitivity of LAMP, PCR and microscopy were 97.47, 97.47, and 61.61%, respectively, and specificity of LAMP, PCR and microscopy were 60.31, 74.60 and 100% respectively. Higher sensitivity of diagnostic test will be more useful in high TB endemicity area to cover most of TB patients under National Tuberculosis Control Programmae. This LAMP assay may be a viable alternative of costly PCR diagnosis. This test may be adopted as supplementary diagnostic test with acid fast smear microscopy for routine diagnosis of human TB. Source

Ramane S.,Mycobacteria Laboratory | Verma R.,Center for Animal Disease Research and Diagnosis | Mondal T.,Mycobacteria Laboratory | Upmanyu V.,Indian Veterinary Research Institute
Journal of Pure and Applied Microbiology | Year: 2014

Mycobacterium bovis 3/86 strain isolated from cattle was characterized based on RD region encoded Mb3904, Mb3905 and Mb2002c gene sequences. PCR was performed to amplify Mb3904, Mb390S and Mb2002c genes. Restriction enzymes digested amplified geneswere cloned in compatible pET vector and sequenced with vector specific primers. The sequenced genes and its deduced amino acid sequences were compared with the published sequences of reference strains. The sequences of the Mb3904, Mb3905 and Mb2002c genes share 99.6 to 100% nucleotide homology and 99.5 to 100% deduced protein sequence homology for all studied genes with published reference mycobacterial strains indicating their conserved nature. Source

Prakash C.,Center for Animal Disease Research and Diagnosis | Das P.,IVRI | Sunil Kumar B.V.,GADVASU | Singh V.,National Research Center on Mithun | And 2 more authors.
National Academy Science Letters | Year: 2015

Present study evaluated nucleotide sequence variability in 3′ end of insertion sequence IS407A in B. mallei NCTC 3709 strain and compared with other B. mallei strains reported worldwide. This insertion sequence IS407A was found conserved among all B. mallei strains studied while it was found absent in B. pseudomallei. Flagellar gene (motility gene) showed single nucleotide polymorphism in B. mallei strains. This insertion sequence can be used as a molecular signature for B. mallei organism and it can be successfully used in development of multiplex PCR for simultaneous detection of multiple equine pathogens like B. mallei (glanders), B. pseudomallei (Melioidosis) and Streptococcus equi (strangles). © 2014, The National Academy of Sciences, India. Source

Khan F.A.,University of Wisconsin - Madison | Das G.K.,University of Wisconsin - Madison | Pande M.,University of Wisconsin - Madison | Singh R.,Center for Animal Disease Research and Diagnosis | Ghosh S.K.,Indian Veterinary Research Institute
Buffalo Bulletin | Year: 2013

Histological examination of H&E stained sections of ovaries collected from cyclic and acyclicbuffaloes (n=6/group) was done in order to evaluate the degree of follicular atresia. The percentages of healthy and atretic follicles were different (P<0.05) between cyclic (53.1% and 46.9%, respectively) and acyclic (10.7% and 89.3%, respectively) buffaloes. Electrophoretic patterns of follicular fluid proteins, studied by SDS-PAGE of follicular fluid samples aspirated from small (5.0- 6.9 mm), medium (7.0-9.9 mm) and large (≥ 10 mm) follicles of cyclic and acyclic buffaloes (n= 6/group), did not reveal any apparent differences between the groups. DNA fragmentation patterns evaluated by using DNA isolated from the cell pellets obtained after centrifugation of the follicular fluid samples from small-, medium- and largesized follicles of cyclic and acyclic buffaloes (n=5/ group) showed a typical apoptotic oligonucleosome ladder pattern in the acyclic group. In conclusion, these results indicate an increased rate of follicular atresia without any qualitative changes in the follicular fuid proteins during ovarian acyclicity in buffalo. Source

Telang A.,Center for Animal Disease Research and Diagnosis
Environmental Toxicology | Year: 2015

This study was undertaken to investigate the toxic effects of imidacloprid (IM) on male reproductive system and ameliorative effect of curcumin (CMN) in male Wistar rats. For this purpose, IM (45 and 90 mg/kg, body weight) and CMN (100 mg/kg, body weight) were administered orally to the rats either alone or in combinations for a period of 28 days. At the end of experiment, male reproductive toxicity parameters (total sperm count and sperm abnormalities), testosterone level, steroidal enzymatic activity [3β-hydroxysteroid dehydrogenase (3β-HSD) and 17β-HSD], and oxidative stress indicators were estimated in testis and plasma. IM treatments resulted in significant decrease (p<0.05) in total epididymal sperm count, sperm motility, live sperm count, and increase (p<0.05) in sperm abnormalities. Activities of gamma-glutamyl transpeptidase, lactate dehydrogenase-x, and sorbitol dehydrogenase were significantly increased (p<0.05), while, 3β-HSD and 17β-HSD enzymatic activity along with testosterone concentration in testis and plasma were decreased significantly (p<0.05) in IM-treated rats. IM exposure resulted in significant increase (p<0.05) in LPO and decrease (p<0.05) in GSH level along with decreased activities of CAT, SOD, GPx, and GST. IM-treated rats showed histopathological alterations in testis and epididymis. However, the reproductive toxicity parameters, oxidative stress indicators, and histopathological changes were minimized and functional restorations were noticed by co-administration of CMN in IM-treated rats. The results of this study suggest that IM-induced male reproductive toxic effects could be ameliorated by CMN supplementation. © 2015 Wiley Periodicals, Inc. Source

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