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Aim:The aim was to determine hemato-biochemical changes and rapid diagnosis of Theileria annulata in naturally infected crossbred cows.Materials and Methods:Blood samples from lactating crossbred cows (n=40) between 3 and 7 years of age and showing clinical signs of tropical theileriosis were collected, with or without anticoagulant, and analyzed for tropical theileriosis by direct smear, direct blood polymerase chain reaction (PCR) detection of merozoite-piroplasm surface antigen (Tams1) gene specific amplicon, estimation of hematological and biochemical parameters. Healthy crossbred cows (n=6), examined free from hemoprotozoan infections were included as control.Results:The infected crossbred cows revealed significantly (p<0.001) lower values of total erythrocytic counts (4.46±0.2 × 106/µL), hemoglobin (Hb 6.025±0.39 g%), packed cell volume (17.05±1.1%), mean corpuscular volume (37.94±1.70 fL) and mean corpuscular Hb (13.5±0.48 pg; p<0.002) compared with healthy control. The serum samples of infected cows revealed profound (p<0.05) hyponatremia (Na 133.21±2.36 mEq/l) and hypocalcemia (Ca 8.39±0.34 mg%). Infected crossbred cows showed a significant increase (p<0.05) of mean serum activity of alanine aminotransferase (61.45±13.36 U/L), aspartate aminotransferase (146.1±20.97 U/L), blood urea nitrogen (28.26±3.90 mg%), creatinine (1.55±0.13 mg%), direct bilirubin (0.33±0.04 mg%; p<0.001) and lactate dehydrogenase (3001.32±167.0 U/L; p<001). Blood direct PCR revealed a 721-bp fragment amplified from the target gene encoding 30-kDa major merozoite surface antigen of T. annulata using specific primer pairs. This assay was positive for all the infected animals.Conclusion:The assessments of hemato-biochemical parameters in T. annulata infected crossbred cows may be useful in understanding disease pathogenesis, prognosis and corrective measures for supportive therapy. Moreover, blood direct PCR can reliably be used for rapid detection of T. annulata in conjunction with microscopic examination.

The present study aimed to diagnose Babesia bigemina in naturally infected crossbred cows and to determine its effect on haemato-biochemical profile of host animals. Blood samples from lactating crossbred cows (n=30) between 3-6 years of age and showing clinical signs of babesiosis were collected, with or without anticoagulant, and analyzed for the protozoa by direct smear, direct blood PCR detection of the apical membrane antigen 1 (AMA-1) gene specific amplicon of B. bigemina and estimation of haematological and biochemical parameters. Healthy crossbred cows (n=10), examined free from haemoprotozoan infections were included as control. Blood Direct PCR revealed a 448-bp amplified fragment. Out of 150 random blood samples screened, (27/150) 18% were positive under light microscope, whereas direct blood PCR revealed (39/150) 26% samples positive for B. bigemina. The result shows higher specificity and sensitivity of PCR test over blood smear examination. The infected group showed significantly (p<0.001) decreased levels of TEC (3.04±0.19), Hb (4.78±0.27) and PCV (14.53 ±0.87) than healthy control animals. However, differences in the red blood cell indices (MCV, MCH and MCHC) were non-significant (p>0.05) between the groups indicating normocytic hypochromic anaemia in affected crossbred cattle. Serum samples of infected cows showed significantly (p<0.01) higher values of ALT (78.83±8.95), AST (146.13±7.62), BUN (27.09±1.02), creatinine (1.93±0.1) and TBIL (1.42±0.06) than that of healthy control. A significant decrease (p<0.01) of TSP (6.12±0.13) and albumin (2.39±0.09) was also recorded in the infected cows compare to healthy control. The standardized blood direct PCR method of the present investigation may be useful for rapid and reliable diagnosis of B. bigemina in conjunction with microscopic examination. Moreover, marked changes in haematological and serum biochemical profile observed in B. bigemina infected crossbred cows may be useful in understanding disease pathogenesis and undertaking necessary corrective measures.

The present study reports the development and use of monoclonal antibody- based latex agglutination test (mAb-LAT) and antigen-detection ELISA (Ag-ELISA) for detection of Trypanosoma evansi infected buffaloes. MAb was produced for the first time against a surface antigen of an Indian isolate of T. evansi and used in development of mAb- LAT and Ag-ELISA for screening 285 buffalo sera samples collected from different regions of Haryana state in India during 2005 and 2006. For comparison, wet blood film (WBF) and microhematocrit technique (MHCT) were also used to detect the parasite in the blood samples from the same animals. Far more buffalo samples were declared positive by mAb-LAT and Ag-ELISA than those by WBF and MHCT. MAb-LAT was found to be a suitable fieldadaptable test for screening of large number of animals for T. evansi infections