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Development And Optimization Of Multiplex Pcr For The Identification Of A, O And Asia 1 Strains Of FMDV In Pakistan

by Muhammad Ikram | Dr. Atif Hanif | Dr. Imran Najeeb | Prof. Dr.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3. Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples. Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l. Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases. Availability: Items available for loan: UVAS Library [Call number: 1189,T] (1).

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Microbial Evaluation Of Raw Meat At Abattoirs And Retail Outlests (Lahore)

by Abid Sarwar | Prof. Dr. Mansur ud Din Ahmad | Dr. Imran Najeeb | Prof. Dr. Azhar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: The objective of this study was to evaluate the microbial quality of meat. The present study was planed to determine the aerobic plate count on meat obtained from the abattoirs and local market. A total of 90 meat samples that were collected for determining the microbiological quality of meat. Half of the meat samples (n=45) were collected from various abattoirs and half of the meat samples (n=45) were collected from retail outlets in Lahore City to get an idea of contamination from slaughtering point to retail outlets. These samples were processed for Aerobic plate counts, E.coli, S.aureus and Salmonella counts. Overall, this study revealed that the level of contamination on meat carcasses was higher in retail meat shops compared to the abattoir. However, the microbial contamination in the abattoir were high if we compare these results to the reports from developing countries like India, Iran and Bangladesh. Bacterial isolates identified and counted from this study were Staphylococcus aureus (44) out of 90 samples was the most abundant as 48.88%, followed by E. coli (43) 47.77% and Salmonella (26) 28.88%. Statistical analysis revealed that analysis of variance between various abattoir and the retail meat shops for E.coli, Salmonella and S.aureus showed significant differences with some exceptions. E.coli counts were significantly higher (P < 0.05) in the meat shops and abattoirs. For E.coli most of the data were significant at 5% level (P < 0.05) with some exception in case of beef and goat samples taken from abattoirs which were non significant because of the unhygienic environments. Analysis of variance for Salmonella between various abattoir and the retail outlets were significant at 5% level (P < 0.05). For S.aureus between various abattoir and the retail outlets showed non significant at 5% level (P > 0.05) with some exceptions in case of beef abattoir and goat retail outlet samples taken which were significant at 5% level (P < 0.05). The higher incidence of microbial load in fresh meat obtained in this study might be attributed to unhygienic and improper handling of animals during slaughter, dressing, evisceration, transportation and unhygienic environments at the retail shops. The usual practice of washing the carcass with the same water in which intestines and offal had been washed was considered as one of the predominant reasons for increased microbial counts of the carcasses. A complete ignorance on the part of the meat handlers/ butchers in hygienic handling of carcasses during slaughter and retailing processes might be the main factors for producing meat with high microbial load. Levels of microbial contamination in Pakistani abattoirs and traditional retail meat shops reflect the hygiene status of meat production in the developing world. Education of the meat retailers' community which runs the traditional meat shops, in terms of the importance of hygienic and sanitary precautions would go a long way towards providing wholesome and safe meat to the consumers. Availability: Items available for loan: UVAS Library [Call number: 1196,T] (1).

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Effect Of Differnet Physico Chemical Substances On The Production Peotential Of Phycocyanin From Spirulina and its Characterization

by Firasat Hussain | Dr. Imran Najeeb | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Spirulina is a multi-cellular, filamentous Cyanobacterium, belonging to a blue-green alga of Cyanophyta. Spirulina is recently proven in animal experiments to exhibit various biological activities such as lowering plasma cholesterol levels and blood pressure. The principal phycobiliproteins present in spirulina are phycocyanin and allophycocyanin which are made up of dissimilar ? and ? polypeptide sub units. The fresh biomass was found suitable for phycocyanin extraction. Freezing and thawing of cells was proved the best method for extraction of phycocyanin (0.4mg/ml), as compared to homogenization, hydrochloric acid and sonication. Nitrogen effects phycocyanin production from spirulina. Different concentrations of nitrogen spirulina medium were provided. Among which 1.875g/L spirulina produced phycocyanin (0.412mg/ml). Phosphate effects phycocyanin production from spirulina. Different concentrations of phosphate spirulina medium were provided.Among which 1.5g/L spirulina produced phycocyanin (0.354mg/ml). There is also effect of temperature on phycocyanin production. Spirulina medium 0.192mg/ml at 25oC, 0.390mg/ml at 30oC, 0.184mg/ml at 35oC. There is also effect of light on phycocyanin production. 0.361mg/ml were produce at 1500 Lux. Molecular weight (66kDa) of phycocyanin was confirmed by SDS-PAGE and explored potential production of phycocyanin from indigenous spirulina. Availability: Items available for loan: UVAS Library [Call number: 1204,T] (1).

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