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Characterization of cholinesterase activities for pesticide exposure in food animalsAbass Askar, Kasim Sakran January 2012 (has links)
The primary aim of the work described in this thesis is to establish a foundation for the applicability of a biochemical biomarker, cholinesterase (ChE) activity in food animal species, as an instrument for evaluating exposure to pollutants as well as predicting high-level effects on public health. Secondary aims are to increase the awareness of pesticide users of anti-ChE exposure, to decide whether poisoning episodes involve anti-ChE by measuring residual effects in tissues, and to identify sources of contamination in food animal tissues. The ChE are specialized carboxylic ester hydrolases that break down esters of choline. They are classified as either acetylcholinesterase (AChE) or butyrylcholinesterase (BChE). Both AChE and BChE activities were found to be higher in cattle than in sheep and higher in erythrocytes than in plasma and serum. The anticoagulant heparin significantly affects AChE activity in plasma compared with EDTA. Of the different tissue tested, the mean of ChE activities was found to be highest in tissue from the liver, followed by lung, muscle, kidney and heart for sheep and cattle. In pigs, the ChE activities tested higher in kidney, liver, lung, muscle and heart. The effect of freezing on ChE activities in liver and muscle tissues was significant inhibition after 6 months at -80 °C, whereas decreased after 3 months at -20 °C. A technique to improve the purification of AChE in sheep tissue was developed. BW284c51 strongly reduced acetylthiocholine iodide (AcTChI) and propionylthiocholine iodide (PrTChI) hydrolysis and slightly affected that of butyrylthiocholine iodide (BuTChI) in the liver, while iso-OMPA had no significant effect for muscle BuTChI of sheep and pigs. Histochemical study of liver tissue found AChE localised mainly in the cytoplasm of the cell lining in the sinusoids. The optimal pH values of AChE and BChE in liver and muscle ranged between 7.8 and 8.5. Both AChE and BChE activities increased when increase the time course and temperature. The half maximal inhibitory concentration (IC50) was found to be higher for carbaryl than dichlorvos (DDVP) and diazinon (DZN). Very little residual AChE activity was seen in the liver, but more was found in muscles. In general, the rate constants of inhibition (ki) values for liver and muscles were increased in different pHs according to the rank order of 8.5 > 7.5 > 6.5, while in plasma it was decreased in different temperatures as follows: 20 °C > 30 °C > 40 °C. The final experiments were carried out at the rate of spontaneous reactivation (ks) of inhibited AChE by DDVP and DZN from liver and muscle was found to be higher in sheep compared to cattle and pig, while the aging of phosphorylated AChE (ka) was found to be higher in cattle compared to sheep and pig. In addition, this study indicated that the developed bispyridinium symmetric (K048) oxime seems to be promising reactivated to DDVP-inhibited AChE for sheep and pigs while HI-6 was effective in cattle.
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Applications of spice extracts and other hurdles to improve microbial safety and shelf-life of cooked, high fat meat products (doner kebab)Al-Kutby, Sahar January 2012 (has links)
There is a growing demand for safe and convenient meat products. The effect of natural spice extracts incorporated with other hurdles for controlling pathogenic bacteria and extending the shelf life of RTE doner kebab were investigated. A comprehensive literature review was undertaken to establish the status of microbial risk, use of additives, knowledge on oxidative deterioration and HACCP associated with meat products. The in vitro antioxidant and antibacterial activities of spice extracts were screened and compared. Cinnamon, clove, and sumac alcoholic extracts demonstrated strong antimicrobial effect, however, rosemary proved effective as antioxidant in a lamb fat model. An accelerated shelf life study on a model system indicated that storage temperature was the most critical factor affecting lipid oxidation, which was effectively delayed by vacuum packaging and rosemary extracts. The effects of spice extracts, packaging and storage time on physiochemical, microbiological, and sensory attributes of doner kebab were evaluated. Application of rosemary and cinnamon extracts significantly reduced TVC, inhibited LAB, and retarded lipid oxidation rate. Sensory evaluation by a consumer panel indicated that only taste and spiciness perception was significantly different between treatments. A challenge test against Listeria monocytogenes showed significant differences between control and spice treatments at day 28. Strong inhibitory effects were associated to high levels of cinnamon particularly when applied after cooking. The effect of heat treatment and sumac (Rhus coriaria) on Bacillus cereus and Clostridium perfringens inactivation was evaluated on a doner kebab prototype. Addition of sumac significantly reduced D-values and z-values for both organisms in comparison to the control. The investigation of the effect of spice extracts, and environmental conditions on changes in growth kinetic parameters for L. monocytogenes and Salmonella Typhimurium showed that spice extracts are highly significant. For both microorganisms, Mumax was reduced as salt and spice concentrations increased, and pH levels decreased. This study shows that spice extracts incorporated with other hurdles can help to maintain safe and good quality RTE doner kebab.
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