Aim:This study was taken up to develop an indirect enzyme-linked immunosorbent assay (i-ELISA) for screening antibodies against Leptospira spp. in canines.Materials and Methods:An i-ELISA was developed using outer membrane protein extracted from Leptospira interrogans serovar canicola used for coating the well with concentration of 0.5 µg/µl. A total of 250 serum samples from clinically affected and apparently healthy dogs were collected along with relevant epidemiological data at Teaching Veterinary Clinical Complex, Veterinary College and Research Institute, Namakkal, and subjected to i-ELISA.Results:Out of 250 sera samples, 140 (56.00%) were found to be positive by i-ELISA. All the sera samples were subjected to microagglutination test (MAT) with panel of 12 different serovars. A total of 71 (28.40%) sera samples were positivity to MAT excluding the sera samples positive to L. interrogans serovars canicola and icterohaemorrhagiae in vaccinated dogs. Sensitivity and specificity of i-ELISA were higher in compared with MAT was 91.54% and 58.10%, respectively.Conclusion:An indirect ELISA developed for the detection of canine antileptospiral antibodies proved to be highly sensitive, rapid and easy to perform and overcome the drawbacks of MAT.
Leptospirosis is a reemerging and a complex zoonotic bacterial disease, caused by pathogenic serovars of Leptospira interrogans. A total of 124 sera samples of dogs belonging to different categories like vaccinated, unvaccinated-semiowned, and stray dogs were subjected to sampling. Microscopic agglutination test (MAT) was conducted by using Leptospira culture. Out of 42 vaccinated dogs, 24 (57%) were positive to one or more serovars. Of the 24, 22 (52.3%), 11 (26.19%), 4 (9.5%), 1 (3%), and 2 (4.7%) were positive to icterohaemorrhagiae, canicola, pomona, grippotyphosa, and autumnalis, respectively. Of the 48 unvaccinated semiowned dogs, 10 (28.8%) showed positive agglutination to one or more serovars. Of the 10 samples, 7 (14.5%), 2 (4.1%), 3 (6.2%), 3 (6.2%), and 5 (10.2%) were positive to icterohaemorrhagiae, canicola, pomona, grippotyphosa, and autumnalis, respectively. Among the 34 stray dogs, 12 showed positive agglutination to one or more leptospiral antibodies. Of the 12 samples, 6 (17.6%) showed positive agglutination to icterohaemorrhagiae, 2 (5.8%) to canicola, 5 (14.7%) to pomona, 7 (20.5%) to grippotyphosa, and 5 (4.7%) to autumnalis. This study emphasized the changing trends in the epidemiology of leptospirosis with higher prevalence of serovar L. grippotyphosa in street dogs.
A cross-sectional study was carried out to identify risk factors for bovine viral diarrhea virus (BVDV) infection in 62 randomly selected dairy herds which were tested for BVD serum antibodies by using an indirect ELISA kit (IDEXX). Results from the chi-square test analysis were interpreted by analyzing by chi-square test. A sum of 500 sera samples were screened and 66 animals (13.20%) showed positive for BVDV antibody. Within herd, BVD seroprevalence was 12-65%. This study concluded that epidemiological risk factors like location, herd size, housing patterns like, tail to tail system, roofing pattern, distance between the manure pit and farm, and distance between farms were significantly associated with BVDV serological status (P < 0.05).
An attempt had been made in this study to know the ability of Indian street rabies virus isolates to induce apoptosis in murine neuroblastoma cells. Thirty Indian street virus isolates were tested by fluorescent antibody technique (FAT) among which 20 samples was positive. These positive samples were used to infect murine neuroblastoma (MNA) cells. Four street rabies virus isolates and challenge virus strain (CVS) were used in this study to show their ability to induce apoptosis. Total RNA was isolated and cDNA was synthesized by using gene specific primers pertaining to Caspase-1, BAD (Proapoptotic gene) and Glycoprotein gene of rabies virus. Challenge Virus Standard has been used as control in the amplification as well as in the demonstration of apoptosis. The expression of Caspase-1, BAD and glycoprotein genes in the murine neuroblastoma cells in comparision with CVS was measured by using densitometer. Street virus isolates expressed higher levels of Caspase-1 and BAD in comparison with CVS. Further TUNEL staining method showed higher apoptotic index when compared to CVS. This study is an initial attempt to exhibit the ability of Indian rabies street virus isolates to induce apoptosis in murine neuroblastoma cells (in vitro).
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