Buffaloes were screened for theileriosis by routine microscopic examination and also subjected for characterisation by PCR technique. Blood samples were collected from lactating buffaloes in post partum period from endemic areas of Athagarh block of Cuttack district, Odisha, India. Genomic DNA of Theileria piroplasm was isolated and genus specific primers were used for amplification of small subunit ribosomal RNA sequences. The amplified PCR products of Theileria spp. were sequenced. Out of 86 cases examined, 21 and 31 samples were found positive by Giemsa stained blood smear method and PCR technique respectively. The PCR product was sequenced and analysed for homology. The identified nucleotide sequence had close sequence homology with Theileria orientalis and Theileria buffeli. These findings also support the fact that 18S small subunit rRNA gene is hyper variable among the species. The nucleotide sequence was submitted to NCBI and a new accession number (MN262069) was assigned.

Raw milk samples of 503 individual buffaloes were collected from dairy farms located in Agra and Mathura cities in North India. Multiple tests (Indirect Fluorescent Agglutination test (i_FAT), IS900 PCR, Microscopy, Indigenous ELISA kit (i_ELISA), Dot-ELISA (d_ELISA) and Latex agglutination test (LAT)) based bio-load and bio-type profile of Mycobacterium avium subspecies paratuberculosis (MAP) was studied. Cumulatively average bio-load was 61.2 percent using three antigen and three antibody based. In i_FAT, IS900 PCR and microscopy, 43.5, 13.3 and 40.9 percent milk were positive for MAP, respectively. Whereas, 32.8, 49.3 and 44.1 percent milk samples were positive in i_ELISA, d_ELISA and LAT, respectively. Bio-typing of representative milk samples using IS900 PCR positive raw milk (67), 13.4 percent were infected with 'Indian Bison Type' biotype using IS1311 PCR_REA. Study concluded that 'Indian Bison type' was the predominant bio-type infecting lactating buffaloes of this region. Raw milk was highly convenient sample in buffaloes and 'milk samples' were first time screened without initial processing of milk samples. Detection limits of each tests was improved. Results of five tests (d_ELISA, LAT, i_ELISA, microscopy, i_FAT were comparable, except IS900 PCR. High bio-load of MAP in milk of buffaloes was major health hazard for human health. High bio-load of MAP was alarming and calls for initiation of Johne's disease control programs in the country.

Tropical theileriosis poses major threat for buffaloes causing significant economical loss to livestock farmers. Early detection and prompt treatment helps to minimise mortality and economical loss. This study was conducted on 79 female buffaloes presented with the signs suggestive of theileriosis. Evaluation of PCR for detection and buparvaquone for efficacy was undertaken. Overall prevalence of T. annulata infection recorded was 22.78 percent by PCR. Adult buffaloes showed higher prevalence (13.92 percent) compared to young buffaloes (8.86 percent). Blood smear examination revealed 38.89 percent sensitivity in detection of Theileria piroplasms. Haematological observations showed significant decreased values of Hb, TEC, PCV and TLC. Neutropenia, monocytopenia, eosinopenia and lymphocytosis were recorded. Buparvaquone was 100 percent effective in complete elimination of T.annulata in infected buffaloes.

A 7 years old female Murrah buffalo was presented to the Referral Veterinary Polyclinic, Indian Veterinary Research Institute, Izatnagar with a history of high fever, anorexia and respiratory distress since 2 days. On clinical examination, high body temperature, congested conjunctival mucous membrane, open mouth breathing, tachypnoea, tachycardia and lymphadenopathy noticed. Clinical pathology revealed leukocytosis, neutrophilia with shift to left and blood sample was found to be negative for haemoprotozoan infection. Bacteriological culture of blood sample revealed mucoid dew drop colonies suggestive of Pasteurella spp. and on Gram's staining of bacterial culture, Gram-negative cocco-bacilli organisms were detected. Further, the results of Pasteurella multocida species specific-PCR (polymerase chain reaction), Pasteurella multocida multiplex capsular PCR typing and Pasteurella multocida serotype B specific PCR revealed that the isolate was of Pasteurella multocida serotype B: 2. ABST (Antibiotic sensitivity test) revealed that the organism was highly sensitive for antibiotic Ceftiofur. The animal was treated with Inj. Ceftiofur sodium (2.2 mg/kg, IM, SID) and other supportive treatment including anti-pyretics, anti-histamines, multivitamins, rumenotorics and probiotics for 5 days. The animal showed marked recovery after complete therapy.

Successful pregnancy is the outcome of a well coordinated embryo- maternal communication events. Few evidences suggest the role of cytokines signaling pathways as mediators of these communications for establishment of pregnancy. In order to investigate the role of cytokines CCL8 and CXCL10 in embryonic implantation during pregnancy, the present study aimed to develop quantitative real time PCR method based on SYBR Green dye chemistry. Primers were designed for the amplification of CCL8, CXCL10 and GAPDH (endogenous control) genes specific to bovines using Primer 3 software. The amplification products for CCL8, CXCL10 and GAPDH (endogenous control) genes yielded fragments of 388, 151 and 81 bp respectively. Purified PCR Products were used for the generation of standard curve for all the three genes. Six scalars tenfold serial dilutions of every PCR product were performed for amplification of genes by optimized protocol. Each sample was run in triplicate along with a no template control for every assay. Each run was completed with a melting curve analysis to confirm the specificity of amplification and lack of primers dimers. The standards generated linear relationships with regression coefficients: r sup(2)=0.996, 0.993 and 0.992 for CCL8, CXCL10 and GAPDH genes respectively. The method posed to be reliable approach for estimating the relative expression of cytokines CCL8 and CXCL10 in peripheral blood leucocytes in buffaloes.

Foot and mouth disease is among the top listed livestock diseases causing severe economic losses. The aim of the present study was to determine the seroprevalence of FMD and detection of FMD virus shedding in milk of apparently healthy buffaloes and cattle of Pakistan. A total of 30 dairy farms were selected and registered in rural areas of Punjab consisting of minimum 15 animals. A total of 180 serum samples were collected and subjected to Non structural proteins (NSP) ELISA. The milk of sero-positive animals were collected and detected for the presence of FMD virus using reverse transcriptase PCR (RT-PCR). The results of current study showed overall seroprevalence 71.66 percent (129/180) with 65.38 percent in buffaloes and 76.47 percent in cattle. The FMD virus was detected in 24.03 percent (31/129) of sero positive samples. Among the FMD virus positive samples 65.51 percent belongs to serotype O and 35.48 percent belongs to Asia I, while none of the other serotypes were detected. The detection of FMD virus from the milk of apparently healthy buffaloes and cattle is an alarming situation and it may be considered as a potential role in the transmission of FMD.

Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne's disease, a disease with considerable economic impact on dairy buffalo herds. The present study was undertaken to assess the prevalence of MAP infection in buffaloes and its excretory pattern through buffalo milk. A total of 74 milk samples were collected from apparently healthy buffaloes of organized and unorganized sectors located in Tamil Nadu and subjected to Ziehl-Neelsen staining and Polymerase chain reaction (IS900 and F57 genes). Out of 74 samples, 3 (4.1 percent), 21 (28.4 percent) and 14 (18.9 percent) samples shed MAP organism by Ziehl-Neelsen staining, IS900 PCR and F57 PCR respectively. Besides age of the animal, stage of lactation and herd management were associated with excretion of MAP in milk. These results showed the high prevalence of MAP infection in buffaloes and warrants further studies and necessary actions to delineate the MAP infection in buffalo population.