Graduate Thesis Or Dissertation
 

Histochemical study of some enzymes of the common liver fluke, Fasciola hepatica L.

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  • Acid and alkaline phosphatase was localized histochemically in the intestinal epithelium and excretory duct respectively in fresh frozen sections of Fasciola hepatica. Polyphenol oxidase was localized in the vitellaria, vitelline ducts and egg-shells. The distribution and nature of esterases was examined histochemically. Acetyl-cholinesterase was observed in the tegument, neural ganglia, neural commisure, acetabulum and oral sucker. Pseudocholinesterase was observed in the pharynx, cirrus, cirrus pouch wall, prostate gland sperm sac, testes, ovary, vitelline duct and duct walls of Mehlis' gland. Use of differential inhibitors of mammalian cholinesterases indicated that presence of acetylcholinesterase in structures containing pseudocholinesterase could not be demonstrated by differential enzyme activity. Non-specific esterase is reported in the excretory ducts and intestinal epithelium. The possible functions of the various esterases is discussed. Succinic dehydrogenase was observed in all those organs and tissues reported to contain esterases. It is reported from the cirrus, cirrus pouch wall, prostate gland and sperm sac for the first time. The effects of various concentrations of Ruelene (4-tert-butyl-2-chlorophenyl methyl methyl phosphoramidate), Halaxon (di(2-chloroethyl) 3-chloro-4 methylcoumarin-7-yl) phosphate, CoRal (0, 0-diethyl-0-3-chloro-4-methyl-2-oxo-2H-l-benzyopyran-7-yl phosphorothioate), Dibenzylamine (DABA, 1, 6 bis (methylamino-methylphenoxy) hexane dihydrochloride), Tris (p-aminophenyl) carbonium chloride (TAC, pararosaniline chloride), Bayer 9015 (B9015, 3, 3'-dichloro-5, 5'-dinitro-0, 0'-biphenol), Bithionol (2, 2'-thiobis (4, 6-dichlorophenol) 'Actamer') were examined for inhibitory effect against fluke cholinesterases. Bithionol and B9015 failed to inhibit esterases activity. CoRal, Neguvon, TAC and Halaxon inhibited all cholinesterase activity. Ruelene reversibly inhibited pharyngeal and reproductive complex cholinesterases but non-reversibly inhibited tegumental and neural Acetylcholinesterase. Dibenzylamine reversibly inhibited all cholinesterases. Bayer 9015 and DABA failed to inhibit excretory duct alkaline phosphatase activity. Bayer 9015 failed to inhibit acid phosphatase activity in intestinal epithelium. Bithionol, Bayer 9015 and Hexachlorophene (2, 2' dihydroxy-3, 5, 6-3'5'6' hexachlorodiphenyl methane) all inhibited succinic dehydrogenase at all concentrations tested. Dibenzylamine had no inhibitory effect while TAC partially inhibited the staining reaction. Incubation of adult flukes in DABA-Hedon-Fleig saline for 8, 18 or 24 hours resulted in a decrease in PAS positive material and loss of tegument. This is compared with Bayer 9015 induced necrosis and a hypothesis explaining the mechanism of this necrosis is presented.
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