Hematobiochemical alterations and direct blood polymerase chain reaction detection of Theileria annulata in naturally infected crossbred cows.

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 × 10(6)/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.

Visceral schistosomiasis of domestic animals in India: humoral immune status of infected cattle, sheep and goats against major polypeptide antigens of Schistosoma indicum and S. spindale.

Polypeptide profiles of Schistosoma indicum and S. spindale adult worm homogenates were obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Humoral immune status of infected cattle, sheep and goats against Schistosoma indicum and S. spindale Ags was determined by immunoblot analysis and by indirect ELISA using four major polypeptides of approximate molecular masses 45 kDa, 40 kDa, 28 kDa and 15 kDa electro-eluted from the gel slices. Cattle sera samples had higher levels of antibodies against Si/s40 and Si/s28 than against Si/s45 antigen. Reasons have been discussed for the absence of detectable levels of anti-Si/s28, -Si/s45 and -Si/s40 antibodies in a significant number of sera samples from S. indicum egg-positive sheep.

Variation in resistance to haemonchosis: selection of female sheep resistant to Haemonchus contortus.

Seventy female lambs (6-7 months old) which were exposed to natural infections of Haemonchus contortus were designated as responders or non-responders on the basis of 10 weekly cumulative faecal egg counts. Selected responder and non-responder lambs were treated with ivermectin, housed separately and 6 weeks post-housing, seven lambs from each group were given a trickle infection of Haemonchus contortus at 1000 L3 daily for 5 days per week up to 2 weeks and examined weekly for 10 weeks after first infection. Analysis of data revealed significantly lower mean faecal egg counts and non-significantly less weight loss in responder than non-responder lambs. Mean values of haemoglobin, packed cell volume, total serum protein and peripheral eosinophil counts were significantly higher in responders than non-responders. In contrast, serum pepsinogen concentration was significantly less in responders than in non-responders. At 10 weeks post-infection, there were fewer pathological lesions and significantly lower worm burdens in responders than in non-responders. These results demonstrate a distinct resistance in responders to Haemonchus contortus infection.

Seasonal distribution of Theileria in Hyalomma anatolicum anatolicum ticks of an endemic area of Haryana, India.

The effect of season on the capacity of Hyalomma anatolicum anatolicum ticks to transmit Theileria was studied by detecting Theileria sporoblasts in the salivary glands of 647 adult ticks moulted in winter (November 1990 to March 1991) and 677 adult ticks moulted in the summer-rainy season (June to August 1991). The intensity (number of infected acini per infected tick) and abundance (number of infected acini per tick examined) of Theileria sporoblasts were significantly (P < 0.05) higher in winter moulted ticks (10.75 and 2.23 respectively) than the summer-rainy season moulted ticks (7.31 and 1.77 respectively). The prevalence of infected moulted ticks was not significantly higher in the summer-rainy season (24.22%) than in the winter (20.71%). A higher percentage of winter moulted ticks had high numbers of infected acini (> or = 11/infected tick) than the summer-rainy season moulted ticks, while the trend was reversed for low numbers of infected acini(5/infected tick), being 47.76% for winter and 73.78% for the summer-rainy season. It was concluded that the winter of Haryana is more favourable to the developing theilerial stages in the ticks than the summer-rainy season.

Folacin and cyanocobalamin in relation to natural Trypanosoma evansi infection in buffaloes.

The haematological values of haemoglobin (Hb), packed cell volume (PCV) and vitamin levels of folacin (folic acid), cyanocobalamin (vitamin B12) of buffaloes, in a surra endemic area of Eastern Haryana, India, were determined. Surra-positive buffaloes had significantly low levels of Hb, PCV, folic acid and vitamin B12. These low levels of folic acid and vitamin B12 may have enhanced the clinical signs of chronic trypanosomiasis, caused by Trypanosoma evansi.

Cell-mediated immune responses to sporozoites and macroschizonts of Theileria annulata.

The nature of cell-mediated immune (CMI) responses was studied in cross-bred bovine calves, immunised by attenuated and allogeneic macroschizonts of Theileria annulata. The CMI responses were also investigated in calves, destined to survive or die of tropical theileriosis (Theileria annulata) induced by a virulent dose of sporozoites or macroschizont-infected lymphoblasts. Calves suffering fatal theileriosis showed poor CMI response. Microcytotoxicity assay revealed an enhanced population of specific cytotoxic cells amongst the peripheral blood lymphocytes (PBL) of calves resolving the infection successfully. The E rosette assay showed proliferation of T cells and the assay for macrophage migration inhibition factor (MIF) demonstrated antigen sensitised cells in the PBL. Calves, immunised by allogeneic and attenuated macroschizont-infected lymphoblasts or those recovering from virulent macroschizont-induced infection, showed protective CMI responses with patterns similar to those appearing after non-fatal sporozoite infection.

Immunoprophylaxis against Theileria annulata with protein from plasma membrane of infected lymphoblasts.

Cross-bred (Bos taurus male x Bos indicus female) calves were protected against the homologous sporozoite-induced challenge of Theileria annulata when immunised with protein from the plasma membrane of macroschizont-infected lymphoblasts of allogeneic origin. However, such protection was parasite-strain specific with the plasma membrane originating from lymphoblasts infected and transformed by the same isolate of T. annulata sporozoites. No protection ensued when the infected lymphoblasts and sporozoites were from immunologically different isolates of T. annulata. There was enhanced proliferation of cells and evidence for lymphokine, originating from antigen-sensitised lymphocytes, demonstrable as macrophage migration inhibition factor in peripheral blood lymphocytes of the calves immunised by plasma membrane protein and challenged by sporozoites of homologous origin.

Epidemiology of gastrointestinal nematodes of sheep and goats in Haryana, India.

Epidemiological studies on some gastrointestinal nematodes of sheep and goats in Karnal, Ambala and Rohtak districts of Haryana (India) revealed that Haemonchus contortus and Trichostrongylus spp. were responsible for parasitic gastroenteritis in these hosts. The adult parasites persisted in the host throughout the year and there was no indication of hypobiosis. The maintenance of the parasite population depended on a continuous cycle of infection between the host and the pasture. The agroclimatic conditions of the districts revealed that, in general, favourable weather conditions for the development and survival of the free-living stages of the parasites existed all year round.