الفهرس يوجد فقط 14 صفحة متاحة للعرض العام
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المستخلص
SUMMARY
This study was divided into two parts according to the area of study;
The first part was done in The Arab Republic of Egypt; a total number of 35 fattening steers (Egyptian native bread cattle) were subjected to slaughtering in Assiut’s slaughterhouses, Assiut Governorate, Egypt. The animals were examined clinically before slaughtering according to Rosenberger (1990). Blood samples were collected in clean dry centrifuge tubes containing disodium salts of EDTA for complete blood picture. Post-mortem examination was carried out and liver tissues specimens were collected and kept in 10 % neutral buffered formalin for histopathological studies.
The second part was done in Japan, with a total number of 90 Japanese Holstein bread cattle of both sexes, were subjected to slaughtering in Asahikawa slaughterhouse, Asahikawa, Hokkaido – Japan. Case history of each animal before slaughtering was studied and then blood samples (whole blood samples, blood serum samples) were collected in clean and dry tubes. After slaughtering each animal was examined for presence of any pathological lesions. Of the total number, 25 animals were clinically healthy, free from any post-mortem pathological lesions and were kept as control, the remaining (35 animals) were found suffering from glycogen degeneration (16), liver abscesses (5), sawdust liver (9) and severe fatty degeneration (5). Liver samples were collected from all animals under study. Blood and liver tissues samples from control animals and those with post-mortem pathological lesions in the liver were subjected to laboratory investigation.
11. Samples
Whole blood
10 ml whole blood were collected in tube containing disodium salt of EDTA and kept directly in cold container containing some ice pieces and used for making the hemolysate according to the following steps:
Blood samples were centrifuged 3500 rpm for 15 min at 4 ºC. Directly after centrifugation, the plasma and buffy coat were drawn off, after that, the packed cells were washed one with ten volumes of cold saline. The packed erythrocytes were divided into two parts in sterile ependorff tubes. One part was used for determination of erythrocytic GSH-Px activity where the RBCs were hemolysed by adding 4 volumes of cold deionized water. The second part was used for determination of G6PD activity of erythrocytes (RBCs were hemolysed by mixing 0.05 ml of the washed cell suspension with 0.5 ml of lysing solution (0.02 % digitonin containing NADP, 15 µmol/l; 16 mg digitonin was dissolved in 80 ml deionized water, filtered through Whatman # 1filter paper and then 1 mg NADP was added) and then the hemolysates were stored at –70 ºC till subjected to analysis.
Intracellular erythrocytic G6PD (U/ g Hb.) activity was measured spectrophotometerically according to the method described by Deutsch (1983). Intracellular erythrocytic GSH-Px (U/ g Hb.) activity was determined by using test kits supplied Oxis Research (Bioxytech GSH-PX-340) Portland.
Serum samples:
10 ml blood samples were collected in clean and dry centrifuge tubes from all animals under study for obtaining serum samples which used for measuring blood serum levels of total protein, albumin and total bilirubin and serum activities of aspartate amino transferase (AST), adenosine deaminase (ADA), glutamate dehydrogenase (GLDH) and γ- glutamyl transferase (GGT) by using Automatic Autoanalyzer (Hitachi Autoanalyzer 7060, Hitachi Ltd. Tokyo, Japan) and test kits supplied by Roche diagnostics.
Liver tissues specimens
Liver samples were collected from each animal under study and divided into two parts; one part (1 cm x 1.5 cm x 5 mm thickness) were preserved in 10 % neutral buffered formalin till subjected to histopathological examination, the other part was preserved directly after collection in ice till preparation; liver tissues specimens were homogenized in four volumes of cold buffer (Tris- HCL, pH 7.5) and then centrifugated at 10000 rpm for 30 min. at 4 ºC, the supernatant fluid containing the enzyme was removed and kept in ice if they will be assayed the same day, otherwise stored at – 70 ˚C and used for measuring hepatic G6PD and GSH-Px activities.
Hepatic G6PD activity (U/ g protein) was measured spectrophotometerically according to the method described by Deutsch (1983). Hepatic GSH-Px activity (U/ g protein) was determined by using test kits supplied by Oxis Research (Bioxytech® GSH-PX-340) Portland.
The classification of different types of liver affections was done according to the gross and histopathological findings; only chronic localized cholangitis was detected in that group of animals examined in the Arab Republic of Egypt. However, glycogen degeneration (16), Liver abscesses (5), sawdust liver (9) and severe fatty degeneration (5) were subjected to study in Japan
Clinical findings:
There were no abnormal clinical signs in animals suffered from glycogen degeneration, chronic localized cholangitis and sawdust liver. However, animals with liver abscesses showed loss of body weight. On the other hand, animals with severe fatty degeneration showed emaciation, weakness, dullness, roughness of coat and pale mucous membranes.
Histopathological findings:
Histopathological examinations of the investigated liver affections revealed increased portal C.T. which was dense, chollagenous and contained few chronic inflammatory cells and bile duct proliferation in cases of chronic localized cholangitis, irregular clear spaces in the cytoplasm of the hepatocytes in cases of glycogen degeneration, focal area of necrosis with neutrophilic infiltration in cases of sawdust liver, round fat globules in the hepatocytes that displace their contents to one side in cases of severe fatty degeneration.
Hematological and biochemical findings:
Hematological findings:
Cases with chronic localized cholangitis showed insignificant change in red blood cell picture, however, there were significant increase in total leucocytic counts with neutrophilia and eosinophilia.
Biochemical findings
Glucose 6 phosphate dehydrogenase activity in the erythrocytes and in the hepatic tissues:
Hepatic G6PD activity showed significant increase in cases of liver abscesses, sawdust liver and severe fatty degeneration. However, there was insignificant change in its activity in cases of glycogen degeneration. There were insignificant changes in erythrocytic G6PD activity in cases of glycogen degeneration, liver abscesses, sawdust liver and severe fatty degeneration.
Glutathione peroxidase activity in in erythrocytes and in hepatic tissues:
GSH-Px activity showed insignificant change in the erythrocytes in cases of glycogen degeneration, liver abscesses, sawdust liver and severe fatty degeneration. However, there was a significant decrease in hepatic GSH-Px activity in cases of severe fatty degeneration.
Total protein and its fractions:
There were insignificant changes in total protein and its fractions in cases of liver abscesses, sawdust liver and glycogen degeneration. However, there were significant decreases in total protein and albumin levels in cases of severe fatty degeneration.
Blood serum level of total bilirubin and serum enzyme activities:
There were significant increases in total bilirubin levels and in AST, γ- GT, ADA and GLDH activities in in cases of glycogen degeneration, liver abscesses, sawdust liver and severe fatty degeneration.