الفهرس 
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Abstract
Sarcocystosis is a zoonotic disease in domestic animals caused by Sarcocystis species, a cyst-forming coccidian parasite with obligatory two host life cycle involving carnivorous as definitive hosts and herbivorous or omnivorous as intermediate hosts. Sarcocystis species were infested a vast range of animals as well as human beings. Found frequently in animal carcasses at slaughter, undermining their value, they have also been found associated with clinical disease. Infestation in food animals could result in loss of weight, poor feeding efficiency, anorexia, fever, anaemia, muscle weakness, reduced milk yield, abortion and death in cases of very severe infection depends on the dose of ingested sporocysts and the immune status of the host.Although buffaloes meat is one of the most important sources of animal protein for a wide range of consumers, but unfortunately there is a wide spread of Sarcocysts in their meat, such infection often results in much economic losses in addition to it is possible zoonosis to human. Sarcocysts in buffaloes meat cause serious changes which may greatly lower the meat quality, lowering its grade and may render it completely unacceptable. As regulations decide, carcasses heavily infected with Sarcocysts are totally rejected, but that of light infection, can pass for human consumption after partial trimming without restriction. This coccidian parasite caused intestinal and muscular sarcocystosis in immunocompetent patients. The serum ALT activity has been regarded as a reliable and sensitive marker of liver disease. ALT may also be a good indicator of overall health, particularly in the context of obesity, the metabolic syndrome, and presence of cardiovascular disease, as many patients affected by these conditions also are at risk of having non-alcoholic fatty liver disease. A growing body of information suggests that determination of AST isoenzymes in human serum is useful in evaluating damage to some of these organs. In hepatic disease, the test is used to assess liver necrosis and for determining prognosis. It may also assist in identifying patients with active alcoholic liver disease. Gamma-glutamyl transferase (GGT) is a second-generation enzymatic liver function test available for several decades, initially used as a sensitive indicator of alcohol ingestion, hepatic inflammation, fatty liver disease, and hepatitis, GGT was associated with an increase in all-cause mortality. Apart from using gamma-glutamyltransferase (GGT) as a predictor of diabetes, cardiovascular and chronic kidney disease, some evidence suggests GGT as an indicator of cancer risks . So in the present study we threw the light on biochemical study of some hepatic enzymes by the effect of buffalo sarcocystosis. Alanine amino transferase, Aspartate aminotransferase and Gammaglutamyl transferase were investigated by the effect of sarcocystosis. Purification, kinetic study and effects of anticiccidial drug (ambrolium) were undertaken for the most significant affected enzyme. Plasma and hepatic samples were collected from 10 control and 15 sarcocystosis naturally infested buffalos. Alanine amino transferase, Aspartate aminotransferase and Gammaglutamyl transferase were investigated in control and sarcocystosis infested plasma and hepatic samples. The results revealed that specific activities of ALT (mU/mg) in different plasma samples from control buffalos ranged from 31 to 57. The specific activities of ALT (mU/mg) in different plasma samples from infested buffaloes ranged from 41 to 87. The Statistical analyses showed that the mean ± SE of specific activities (mU/mg) of ALT in different control plasma samples from buffaloes was 46.33±2.93 while that from infested plasma samples was 56.67±3.74. The specific activities of ALT were non-significantly increased in plasma of infested buffaloes (P=0.067). The specific activities of ALT (U/mg) in different hepatic samples from control buffalos ranged from 0.68 to 1.39. The specific activities of ALT (U/mg) in different hepatic samples from infested buffaloes ranged from 0.08 to 0.38. The Statistical analysis showed that the mean ± SE of specific activities (U/mg) of ALT in different control hepatic samples from buffaloes was 1.04±0.076 while that from infested liver samples was 0.21±0.025. The specific activities of ALT were highly significantly decreased in hepatic samples of infested buffaloes (P=0.000). The specific activities of AST (mU/mg) in different plasma samples from control buffalos ranged from 105 to 230. On the other hand, the specific activities of AST (mU/mg) in different plasma samples from infested buffaloes ranged from 120 to 260. The Statistical analysis showed that the mean±SE of specific activities of AST (mU/mg) in different control plasma samples from buffaloes was 161.1±14 while that from infested plasma samples was 178±11.51. There was non-significant increase in specific activities of AST in plasma samples from infested buffaloes (P=0.362). The specific activities of AST (U/mg) in different hepatic samples from control buffalos ranged from 0.56 to 1.86. On the other hand, the specific activities of AST (U/mg) in different hepatic samples from infested buffaloes ranged from 0.69 to 2.73. The Statistical analysis showed that the mean±SE of specific activities of AST (U\mg) in different control hepatic samples from buffaloes was 1.024±0.149 while that from infested hepatic samples was 1.74±0.157. The specific activities of AST were significantly increased in hepatic tissue of infested buffaloes (P=0.005). The specific activities of GGT (mU/mg) in different plasma samples from control buffaloes ranged from 8.6 to 12.1. On the other hand, the specific activities of GGT (mU/mg) in different plasma samples from infested buffaloes ranged from 9.5 to 34. The Statistical analysis of specific activities of specific activities of GGT (mU/mg) in different control plasma samples was 10.37±0.4 while that from infested plasma samples was 16.77±1.98. The specific activities of GGT were significantly increased in plasma of infested buffaloes (P=0.016). The specific activities of GGT (U/mg) in different hepatic samples from control buffalos ranged from 0.59 to 1.24. On the other hand, the specific activities of GGT (U/mg) in different hepatic samples from infested buffaloes ranged from 0.12 to 0.55. The Statistical analysis of specific activities of GGT (U/mg) in different control hepatic samples was 0.85±0.064 while that from infested hepatic samples was 0.30±0.035. The specific activities of GGT were significantly decreased in hepatic of infested buffaloes (P=0.000). The albumin (g/dl) from control buffaloes plasma ranged from 2.7 to 3.59 and that from sarcocystosis- infested plasma ranged from 2.11 to 3.5. The total protein (g/dl) from control buffaloes plasma ranged from 6.39 to 7.2 and that from sarcocystosis- infested buffaloes plasma ranged from 6.03 to 7.2. Statistical analysis of albumin (g/dl) from control buffaloes plasma is 3.1±0.195 while that from sarcocystosis-infested plasma is 2.75±0.243 with a non-significant decrease (P=0.317). Statistical analysis of total protein (g/dl) from control buffaloes plasma is 6.723±0.191 while that from sarcocystosis-infested plasma is 6.546±0.189 with a non-significant decrease (P=0.54). Prothrombin time (sec), activity (%) and INR in different plasma samples from control buffaloes ranged from 14.8 to 20.09, 60 to100 and 1 to 1.55, respectively. On the other hand, prothrombin time (sec), activity (%) and INR in different plasma samples from infested buffaloes ranged from 20.5 to 30.6, 29.5 to 58 and 1.59 to 2.9, respectively. Statistical analysis of prothrombin time from plasma of control buffaloes is 17.25±1.14 while that from sarcocystosis-infested plasma is 23.56±1.89 with a significant increase in prothrombin time (P=0.032). Statistical analysis of prothrombin activity (%) from plasma of control buffaloes is 80.12±8.68 while that from sarcocystosis-infested plasma is 48.1±5.36 with a significant decrease in prothrombin activity (P=0.013). Statistical analysis of International normalized ratio (INR) from plasma of control buffaloes is 1.24±0.12 while that from sarcocystosis-infested plasma is 1.98±0.25 with a significant increase in INR (P= 0.041). The GGT from hepatic samples from control buffalo was purified with a specific activity of 0.034 by DEAE cellulose column chromatography with a yield of 6.08. On the other hand the GGT from hepatic samples from infested buffalo was purified with a specific activity of 0.024 by DEAE cellulose coloumn chromatography with a yield of 5.2. SDS Poly acrylamide gel electrophoresis showed the different steps of purification of hepatic GGT from control and infested buffalos at 10% poly acylamide gel. SDS Poly acrylamide gel electrophoresis showed also purified hepatic GGT (after DEAE cellulose column step) from control and infested buffaloes. The purified hepatic GGT consisted of two subunits with molecular weight of 29.000 and 70.000. The Effect of substrate concentration on purified GGT showed that the values of Km of buffalo GGT from control and Vmax were 4.396 mM and 6802.7 U/mg, respectively while these values were 14.81 mM and vmax was 2020.2 U/mg, respectively for hepatic GGT from infested buffalos. The heat stability of hepatic buffalo GGT showed that with increasing of temperature from 37 to 70 °C the specific activity (U/mg) of control buffalo GGT was decreased from 7.75x10-3 to 0.36 x10-3.The specific activity (U/mg) of infested buffalo GGT was decreased from 7.05 x10-3 to 0.36 x10-3 with increasing of temperature from 37 to 70 °C. The optimum temperature for hepatic buffalo GGT was 37 °C, at which the specific activity of hepatic buffalo GGT from control was 7.75 x10-3 and from infested was 7.05 x10-3. The enzyme from the infested buffalo is less stable at 60 °C compared to that from control. The effect of ambrolium on purified GGT from control and infested buffalos was studied kinetically by Dixon plot. It showed an inhibitory effect of competitive type inhibition as the two lines intersect between the two axes. The kinetic parameters of the effect of ambrolium on purified GGT from control buffalo showed the KI value was 1.42mM while the kinetic parameters of the effect of ambrolium on purified GGT from infested buffalo showed the KI value was 32.23mM. The histopathological examination of control and infested hepatic buffalo samples showed normal structure of buffalo liver while infested hepatic sample showed dilated and degenerated central vein, vacoular degenerative changes in hepatocytes, focal aggregation of polymorph nuclear leucocytes, hyperactivation pkuffer cells , Necrosis, intracellular haemorrage, piknosis of nucleus, eosinophil infiltration, and degenerated and fibrotic bile duct, congestion, haemorrge in hepatic parenchyma, and haemolysis of RBCs, and presence of sarcocystis-schizont stage surrounding areas of necrosis.