tick borne haemoparasites and haemorickettsiales pose a major health risk to animals worldwide. the present study reports the development and validation of multiplex PCr to simultaneously detect the most prevalent tick borne pathogens infecting dogs in kerala, South India. the assay targeting the small subunit ribosomal rNa genes of the organisms could amplify well demarcated amplicons of B. canis vogeli, B. gibsoni and E. canis. In the study population, which included both healthy dogs as well as those with clinical symptoms suggestive of the three infections under study, 46.6% animals were infected with one of the three pathogens, amongst which the occurrence of B. gibsoni was significantly the highest. Natural co-infections were also detected in nine dogs, which suggests the suitability of the assay to assist in the selection of pathogen specific treatment protocols.

Aim:Canine babesiosis is an important vector-borne hemoparasitic disease caused by Babesia canis vogeli and Babesia gibsoni, in India. The communication places on record the salient findings of the study directed to detect and characterize the pathogenic B. gibsoni isolates of Kerala state.Materials and Methods::A total of 150 dogs were examined for the presence of hemoparasites by light microscopy as well as by PCR targeting the 18S rRNA gene of B. gibsoni. Hematological parameters were also analysed. Phylogenetic tree was constructed based on Tamura kei model adopting ML method.Results::A sensitive and specific polymerase chain reaction assay was developed with newly designed primer pair BAGI-F/BAGI-R for the amplification of 488 bp fragment of 18S rRNA gene of B. gibsoni. Out of the 150 dogs examined, molecular evidence of B. gibsoni was recorded in 47.3% animals, while light microscopy detected the infection in 26.67% cases. The phylogenetic analyses revealed that B. gibsoni, Kerala, isolate was closest and occurred together with Bareilly isolate. Anemia and thrombocytopenia were the significant hematological alterations in chronic B. gibsoni infection.Conclusion::A high prevalence of natural infection of B. gibsoni was detected among the study population. The affected animals showed anaemia and thrombocytopenia. Phylogenetic analysis of this pathogenic isolate from south India revealed the closest similarity with Bareilly isolates.

Polymerase chain reaction (PCR) primers specific to exon 2 of the bovine lymphocyte antigen (BoLA)-DRB3 gene were used successfully to amplify the equivalent region in 34 Murrah and 36 Surti buffaloes selected at random. The 304 bp amplified product of the DRB3 gene was separately digested with BstγI, HaeIII and Rsal enzymes. Digestion with BstγI enzyme did not reveal any polymorphism and all animals showed a single restriction pattern, which corresponded exactly to the BstγI pattern ‘b’ previously described for cattle. Digestion with HaeIII enzyme resulted in five patterns, four of which corresponded to the Haelll patterns previously reported in cattle. The new HaeIII pattern was observed in both the breeds of buffaloes studied. The fragment analysis with RsaI revealed 13 different patterns. All of these RsaI patterns corresponded to the RsaI patterns previously described for cattle. The high degree of similarity in the restriction fragment length polymorphism (RFLP) patterns of cattle and buffalo observed in the present study provide evidence for the strong conservation amongst other bovine species, of restriction sites previously reported in cattle.

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The legendary Vechur cattle of Kerala, described as a very short breed, and the crossbred (CB) Sunandini cattle population exhibited great phenotypic variation; hence, the present study attempted to analyze the genetic diversity existing between them. A set of 14 polymorphic microsatellites were chosen from FAO-ISAG panel and amplified from genomic DNA isolated from blood samples of 30 Vechur and 64 unrelated crossbred cattle, using fluorescent labeled primers. Both populations revealed high genetic diversity as evidenced from high observed number of alleles, Polymorphic Information Content and expected heterozygosity. Observed heterozygosity was lesser (0.699) than expected (0.752) in Vechur population which was further supported by positive F value of 0.1149, indicating slight level of inbreeding in Vechur population. Overall, F value was 0.065, which means genetic differentiation between crossbred and Vechur population was 6.5%, indicating that the crossbred cattle must have differentiated into a definite population that is different from the indigenous Vechur cows. Structure analysis indicated that the two populations showed distinct differences, with two underlying clusters. The present study supports the separation between Taurine and Zebu cattle and throws light onto the genetic diversity and relationship between native Vechur and crossbred cattle populations in Kerala state.