HISTOCHEMICAL AND BIOCHEMICAL STUDIES OF BILIARY AMPHISTOMOSIS IN BUFFALOES (Bubalus bubalis) WITH SPECIAL REFERENCE TO CHARACTERIZATION OF Gigantocotyle explanatum ANTIGEN
Loading...
Date
2005
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P
Abstract
ABSTRACT :
The research work on “Histochemical and Biochemical studies of Biliary Amphistomosis in buffaloes (Bubalus bubalis) with special reference to characterization of Gigantocotyle explanatum antigen” was undertaken to carryout the histochemical and biochemical studies of biliary amphistomosis with an attempt to characterize Gigantocotyle explanatum antigen, besides recording the prevalence of biliary amphistomosis in certain places of Andhra Pradesh.
A total of 238 buffaloes were examined in eight (8) places of Andhra Pradesh. Based on slaughter house examination, the per cent prevalence of biliary amphistomosis in buffaloes was 35.71. Among the eight (8) places under this study the prevalence was more in Renigunta (50.00%) followed by 42.10, 37.93, 37.03, 34.61, 32.00, 31.25 per cent at Guntur, Kadapa, Vizag, Vijayawada, Rajahmundry, Hyderabad and was least in Vijayanagaram (27.77%), respectively.
The prevalence of biliary amphistomosis with regard to age, sex, breed and season were also studied.
Among 238 buffaloes examined for biliary amphistomosis, 79 were below 6 months age group, out of which 18 were found positive with 22.78 per cent prevalence. In the age group of 6 months to 5 years, out of 60 buffaloes, 24 were found positive with 40 per cent prevalence and 99 of above 5 years age group, 43 were found positive with the prevalence rate of 43.43 per cent.
Among the total (238) number of buffaloes examined for biliary amphistomosis, 116 were males in which 50 were found positive with a prevalence rate of 43.10 per cent and 122 were females in which 37 were found positive with a prevalence rate of 30.32 per cent.
Among 238 buffaloes examined for biliary amphistomosis, 113 were cross-bred animals, out of which 42 were found positive with a prevalence rate of 37.16 per cent and 120 were non-descriptive, in which 47 were positive (39.16%). Among 238 buffaloes examined for biliary amphistomosis 109, 59 and 70 animals were examined in rainy, winter and summer seasons, out of which 45, 15 and 25 were found positive with a per cent prevalence of 41.28, 25.42 and 35.71, respectively.
Histopathological examination of liver tissue sections in buffaloes naturally infected with biliary amphistomosis, bile ducts revealed thickening and desquammation of the bile ductular epithelium with periductular infiltration of cells. Periductular fibrosis was evident with mononuclear cell infiltration. Mild proliferation of bile duct was also observed. Sinusoidal haemorrhages with dilatation of sinusoids were noticed. Hepatic cells were swollen with granularity in the cytoplasm (cloudy swelling). Thickening of glissions capsule was found.
Histochemical observation of frozen liver tissue sections in buffaloes naturally infected with biliary amphistomosis – alkaline phosphatase and acid phosphatase enzyme activity by Gomori’s method was seen in sinusoidal spaces and hepatic cells. Alkaline phosphatase activity was noticed as black colour where as acid phosphatase activity as brown colour in the stained infected liver tissue frozen sections and correlation with the histological findings of liver necrosis.
In an attempt to estimate the biochemical constituents (total protein, albumin, globulin, A/G ratio, cholesterol, alkaline phosphatase) in the serum samples collected from naturally infected buffaloes, there was a decrease in the total protein, albumin, A/G ratio but slight increase in globulin, cholesterol and alkaline phosphatase levels. Crude antigen of Giganotocotyle explanatum having protein concentration of 10.47 mg/ml was purified by column chromatography with sephadex G -200 using 0.15 M PBS (PH 7.4) buffer. A total of 50 fractions were collected and were monitored for protein concentration using spectrophotometer at 280 nm. The purified fractions showing the maximum peak value F1, F2 and F3 were injected in rabbits to raise the hyperimmunesera and was confirmed by double immuno diffusion test.
The whole homogenate and fractions of Gigantocotyle explanatum were subjected to 5 per cent SDS-PAGE for the analysis of polypeptide profile. SDS-PAGE revealed a total number of 17,16 and 23 polypeptides, respectively. The apparent molecular weight of the polypeptides of these ranged from <14 to 165 KDa.
Description
THESES
Keywords
BILIARY AMPHISTOMOSIS;BUFFALOES; Gigantocotyle explanatum