Qualitative and quantitative assessment of Theileria annulata in cattle and buffaloes Polymerase Chain cattle and buffaloes Polymerase Chaincattle and buffaloes Polymerase Chain Reaction

Bovine tropical theileriosis caused by Theileria annulata is a tick-borne disease associated with high morbidity and mortality in the livestock. The conventional method of diagnosis is by the demonstration of the parasite stages by microscopic examination. This method suffers from low sensitivity, making it even more difficult to detect piroplasms in the carriers. PCR based assays are known to be more sensitive. The present study was undertaken to detect and quantify T. annulata in the blood of clinically infected and carrier animals using a quantitative PCR protocol targeting the gene encoding the major merozoite piroplasm surface antigen Tams 1. A total of 116 samples were collected from infected as well as apparently healthy cattle and buffaloes. Of these, 74 samples (63.79%) were positive for T. annulata by real-time PCR, including the 15 samples that were positive by Giemsa staining. The parasite load ranged from 1.39 x 106 to 3.35 x 109 and 0.35 x 106 to 2.83 x 107 ml-1 of blood in cattle and buffalo samples, respectively by qPCR. Our study suggests that real-time PCR assay can be used to detect and quantify the load of T. annulata in the blood of cattle and buffaloes. It also serves as a support to clinical diagnosis and assessment of carrier status in apparently healthy animals.

Molecular identification of methanogenic archaea from Surti buffaloes (bubalus bubalis), reveals more hydrogenotrophic methanogens phylotypes

Methane emissions from ruminant livestock are considered to be one of the more potent forms of greenhouses gases contributing to global warming. Many strategies to reduce emissions are targeting the methanogens that inhabit the rumen, but such an approach can only be successful if it targets all the major groups of ruminant methanogens. Therefore, a thorough knowledge of the diversity of these microbes in breeds of buffaloes, as well as in response to geographical location and different diets, is required. Therefore, molecular diversity of rumen methanogens in Surti buffaloes was investigated using 16S rRNA gene libraries prepared from pooled rumen contents from three Surti buffaloes. A total of 171 clones were identified revealing 23 different sequences (phylotypes). Of these 23 sequences, twelve sequences (12 OTUs, 83 clones) and 10 sequences (10 OTUs, 83 clones) were similar to methanogens belonging to the orders Methanomicrobiales and Methanobacteriales, and the remaining 1 phylotype (5 clones) were similar to Methanosarcina barkeri. These unique sequences clustered within a distinct and strongly supported phylogenetic group. Further studies and effective strategies can be made to inhibit the growth of Methanomicrobiales and Methanobacteriales phylotypes to reduce the methane emission from rumen and thus help in preventing global warming.

Immunotoxicity induced by subacute acephate exposure in white leghorn cockerels.

Acephate (Ace) is an organophosphate foliar spray insecticide of moderate persistence with residual systemic activity of about 10–15 days. This study was designed to evaluate the immunotoxic effect of oral subacute acephate exposure in 125-day old WLH Cockerel chicks for 28 days. The experimental birds were randomly divided into five groups (C1, C2, T1, T2 and T3), each comprising 25 birds. The birds of group C1 were given no treatment and served as control. Group C2 was administered groundnut oil (1ml/kg) and served as control (vehicle). Group T1 was given ALD50/40 (21.3 mg/kg), and Group T2 was put on ALD50/30 (28.4 mg/kg), while group T3 received ALD50/20 (42.6 mg/kg) of acephate suspended in groundnut oil. Experimental birds of all groups were vaccinated with New Castle disease vaccine at day 7. Blood was collected at two-week intervals for evaluation of humoral immune response. Parameters such as TLC, TP, antibody titre against ND vaccine, DNCB dye test, and histopathology of immune organs were studied to evaluate immunotoxicity. The results were statistically compared (p<0.05) with the control. Acephate produced decreased humoral immune response in terms of New Castle disease vaccine antibody titre, total protein, serum globulin, and serum albumin. Cell mediated immune response was checked with 1-chloro 2, 4 dinitrobenzene dye dermal sensitization test, and it did not reveal any significant differences. Lymphoid organs such as thymus, spleen, bursa, and liver were weighed during necropsy for calculation of organ weight: body weight ratio. After 28 days of acephate exposure, organ:body weight ratios of immune organs were significantly reduced except liver:body weight ratio on 14 days of exposure, which was increased in all treatment groups as compared to control groups. Histopathologically, bursa and spleen showed mild depletion of lymphocytes. To further identify the specific type of cell death as apoptotic or necrotic, DNA ladder assay was performed. DNA fragmentation assay detected ladder pattern (180bp) in DNA from hepatocytes and splenocytes of acephate-treated birds. It is concluded that acephate is immunotoxic, and exerts its immunotoxicity through induction of apoptosis and alteration of immunological parameters.