Slaughterhouse prevalence of ovine paratuberculosis in Southern India.

Ovine paratuberculosis is a threat to small animal holders in terms of economic loss such as reduced growth performance and early culling. In order to study the slaughterhouse prevalence of ovine paratuberculosis, the slaughterhouse sheep samples (which are poor in body condition) collected over a period of two and half years from 1,034 suspected male sheep (poor in body condition) and 40 normal sheep (good body condition and subsequently negative by all the diagnostic tests employed) aged between 16 and 18 months were slaughtered at various abattoirs of Tamil Nadu. All the sheep taken in this study were maintained in almost same management conditions. DNA was extracted from 1,034 intestinal tissue and mesenteric lymph node and 121 were positive by IS 900 PCR. One hundred ten and 56 were positive by absorbed ELISA and Ziehl-Neelsen staining, respectively. In histopathology, 28 animals showed gross lesions of paratuberculosis infection (20-multibacillary and 8-paucibacillary forms). Out of 1,034 sheep tissues cultured, 32 showed cultural growth in Middlebrook 7H9 and 26 in Herrold's egg yolk medium. None of the 40 normal sheep were positive by any of the tests employed. In general, the mean body weight of paratuberculosis-affected animal either by any one of the tests employed was less than the non-affected sheep. The approximate economic loss per sheep/farmer/year is around Rs 1,840 (US$ 38.33) in paratuberculosis-affected sheep.

Detection of rabies virus antigen or antibody using flow cytometry.

BACKGROUND: Rabies is invariably a fatal encephalomyelitis that is considered to be a serious public health problem. Rabies diagnosis must be rapid and conclusive. Detection and quantification of antirabies antibodies is used for assessment of the effectiveness of rabies vaccines. Hence, computer-automated detection of fluorescence using flow cytometry was attempted to reduce the work time required to undertake the conventional rapid fluorescent focus inhibition test (RFFIT). METHODS: Pasteur virus (PV)-infected mouse neuroblastoma (MNA) cells were stained with rabies virus antinucleocapsid antibody, fluorescein isothiocyanate (FITC) conjugate, and the percentage of infected cells at 24, 48, and 72 h postinfection (PI) was determined using flow cytometry. Serum samples containing known antibody titres estimated by RFFIT in terms of IU/ml were used to neutralize 50 FFD50 dose of PV. The percentage of MNA cells infected by the un-neutralized virus was estimated by flow cytometry. Using the value of the percentage of cells infected in the presence of known negative serum as 100%, the infection inhibition caused by antibodies at each dilution of positive reference serum was calculated and a regression equation generated for the prediction of rabies virus antibody titres in test sera samples as equivalent units per ml (EU/ml). RESULTS: There was a significant increase in the percentage of infected cells between 24 and 48 h PI from 26.45 to 75.28%. The percentage of cells having high side scatter was also highest at 72 h PI (11.11%). Antibody titres predicted by flow cytometry and those estimated by RFFIT as IU/ml showed a correlation coefficient of 0.74. CONCLUSIONS: Thus, flow cytometry could be used to detect rabies virus antigen in infected cells and to predict serum antibody titres from a single dilution of serum tested with the potential advantages of automation, rapidity, and lack of subjectivity. It has the potential to replace the time-tested RFFIT in rabies serology in the years to come.