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.

The study describes prevalence, clinical symptoms and risk factors for brucellosis in personnel engaged in veterinary health care in Karnataka, India. A total of 1050 sera samples were collected from animal handlers, veterinarians, veterinary students, para-veterinarians and persons engaged in artificial insemination of animals. The sera samples were tested for brucellosis by Rose Bengal plate test (RBPT), serum agglutination test (SAT), IgG and IgM indirect ELISA and PCR. Age, sex, clinical symptoms and risk factors were recorded in structured questionnaire. Of the 1050 samples tested, 6.76, 6.38, 3.90, 2.67 and 2.0% were positive by IgG ELISA, RBPT, SAT, IgM ELISA and PCR, respectively and overall prevalence recorded was 7.04%. The prominent clinical symptoms observed were intermittent fever (71.62%) followed by joint pain and body aches. A high degree of suspicion, awareness and multimodal diagnostic approach is suggested for early diagnosis, treatment and disease follow up.

Although host specificity has been observed in different species of Brucella, crossing the animal host boundary is likely to occur at any time. In this study, Bruce ladder PCR and abortus–melitensis–ovis–suis (AMOS) PCR assays were used to characterize 47 Brucella isolates from Indian origin in order to know exact species for understanding epidemiology of brucellosis. Out of them, 28, 14, and 5 isolates were found to be Brucella abortus, Brucella melitensis, and Brucella suis, respectively. Further analysis by AMOS PCR has identified that all the B. abortus isolates belong to any one of the biovar 1, 2, or 4; of the five B. suis isolates, three belong to biovar 1 and two belong to any one of the biovar 2, 3, 4, or 5. Although this multiplex Bruce ladder PCR is useful in differentiating all Brucella species, elaborate study is required to further characterize the isolates at exact biovar level.

Background and Aim: The present serodiagnosis of brucellosis in livestock is based on the whole cell or smooth lipopolysaccharide of the Brucella organism in which specificity is hampered by the cross-reactivity, especially with the antibodies against Yersinia enterocolitica O:9 organism. The problem can be addressed by screening for better immunodominant antigens. Hence, the present study was undertaken to screen protein antigens of Brucella abortus for their diagnostic potential in cattle brucellosis. Materials and Methods: Protein antigens of B. abortus (n=10) non-reactive to antibodies against Y. enterocolitica O:9 were selected, expressed in Escherichia coli, assessed the reactivity of expressed recombinant proteins by Western blot, standardized indirect-enzyme-linked immunosorbent assay (ELISA) for detecting Brucella antibodies in cattle serum, and comparative evaluation was done. Results: All the selected protein antigens were expressed and in the Western blot with Brucella antibodies positive cattle serum, six recombinant (Brucella protein 26 [BP26], Cu-Zn Superoxide dismutase [SodC], B. abortus I-1885, Serine protease, Bacterioferritin, and Brucella Lumazine Synthase [BLS]) proteins showed reaction whereas none of the proteins showed reactivity with Brucella negative cattle serum. ELISA has been done using known Brucella positive and negative cattle sera samples (n=113 each) in which the performance of recombinant proteins in diagnosing brucellosis was in the order of BP26 > BLS > SodC followed by rest of the proteins. BP26 based ELISA was found to be better with area under the curve as 0.953, and diagnostic sensitivity, diagnostic specificity, and Youden's index of 90.27%, 95.58%, and 0.8584, respectively, with the excellent agreement (k=0.85). Conclusion: BP26 could be a potential diagnostic antigen among the immunodominant proteins of B. abortus in ruling out Y. enterocolitica O:9 infection while diagnosing brucellosis in cattle herds.

Leptospirosis is a re‐emerging zoonotic disease of animals and humans caused by pathogenic Leptospira, which has major public health concerns. The study is aimed to express the recombinant outer membrane protein (OMP) A‐like protein (rLoa22) and transmembrane (rOmpL37) protein of L. interrogans serovar Hardjo in the E. coli and their evaluation as a diagnostic antigen in the latex agglutination test (LAT) to detect anti‐leptospiral antibodies in the sera of animals. The Loa22 and OmpL37 gene lacking signal peptide coding sequences were individually amplified (522bp and 963bp), by PCR, and directionally cloned into a pETite N‐His Kan vector for expression. The expressed purified proteins were characterized by SDS‐PAGE and immunoblot, which confirmed leptospiral specific reactive protein with a molecular weight of ~19 kDa and 36 kDa, respectively. The sensitized latex beads coated with these OM proteins separately were evaluated in LAT, using cattle sera of microscopic agglutination test (MAT) confirmed positive (n=53) and negative (n=52) cases of leptospirosis. The rLoa22 LAT and rOmpL37 LAT revealed the relative diagnostic sensitivity of 94.34 % & 96.23 %; specificity of 92.31 % & 96.15 %; and accuracy of 93.33 % & 96.19 %, with the excellent agreement of Cohen’s kappa value of 0.87 & 0.92, respectively. After extensive evaluation, this rapid recombinant protein‐based field diagnostic test can be applied as a screening test for the detection of anti‐leptospiral antibodies in the sera of animals in the field conditions.

Background and Aim: Brucellosis is a zoonotic disease of high economic and public health importance in large and small ruminant populations worldwide. A cross-sectional study was conducted to determine the seroprevalence and risk factors of brucellosis in small ruminants in organized farms in the southern region of India. Materials and Methods: Farms exclusively rearing sheep and goats were selected based on the number of animals (small, medium, or large) and the location of the farm (urban, periurban, or rural). A total of 1499 serum samples; 1001 from sheeps and 498 from goats were sourced from six sheep and four goat farms and tested using Rose Bengal Plate and indirect Enzyme-Linked Immunosorbent Assay tests. Results: The apparent prevalence of brucellosis was higher in sheep (8.29%, 95% CI 6.7-10.1) than goats (5.82%, 95% CI 4.0-8.2). The true adjusted population level seroprevalence was also higher in sheep, at 7.7% (95% CI 6.0-9.6) than in goats, at 5.1% (95% CI 3.2-7.6). According to bivariate categorical analysis, six highly significant (p<0.001) animal- and farm-level risk factors for sheep were age, breed, number of lambings, history of abortion, rural farms, and presence of dogs on the farm. In goats, five significant risk factors were found: History of abortion, separate sheds, dogs on the farm, weekly veterinary consultation, and lack of brucellosis awareness. In a logistic regression model, abortion (OR adjusted 10.8, 95% CI 1.2-96.12), rural farms (OR adjusted 8.5, 95% CI 3.6-20.0), and absence of separate sheds on the farms (OR 1.9, 95% CI 1.1- 3.5) were found to be significant risk factors for ovine brucellosis. Conclusion: The use of complementary measures to tackle the multiple animal- and farm-level risk factors may help to reduce the disease burden in the absence of a vaccination policy for small ruminants in India.