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عزل وتشخيص الجين lipA المنتج من بكتريا الزائفة الزنجارية من مياه الصرف الصناعي == Isolation And Identification of lipA Gene Producing Pseudomonas Aeruginosa From Industrial Wastewater
Author name:
انتصار طه لفتة
Supervisor name:
واثق عباس الدراغي
General topic:
Biology
Specific topic:
Biotechnologies
Degree:
Master
University:
University of Baghdad
Language:
Arabic
University location:
Baghdad
First pages:
24T2914 - p.pdf
Abstract:
توضح هذه الدراسة عزل lipA gene من الزوائف الزنجارية من مياه الصرف الصناعية للزيوت النباتية وتشخيصها اعتمادا على تفاعل البلمرة المتسلسل (PCR). استغرقت هذه الدراسة تسعة اشهر, من شهر شرين الاول 2014 لغاية حزيران 2015. تم جمع خمسين عينة من مياه الصرف الصناع | This study clarify the isolation and identification of lipA gene in Pseudomonas aeruginosa from industerial wastewater of vegetable oils factories depending on the polymerase chain reaction (PCR) technique. The present study had taken nine monthes starting from October 2014, till the mid of June 2015. Fifty samples of industrial wastewater were collected from the factories of the general company of vegetable oils, fourty from AL Rasheed factory and ten from AL ameen factory, the samples collected from the physical, chemical and biological treatment units, and other different wastewater tanks departments. While the four sewage samples were collected from sewer service Baghdad /Alrustumaiya. For Identification of Pseudomonas aeruginosa, two types of tests were used in this study. The first type was the routine tests, which include selective cultures, bacteriological and biochemical tests. Thirty four of fifty samples (68%) gave a positive growth and results for tests that were used to confirm the presence of P. aeruginosa. In addition to diagnosis P. aeruginosa in sewage sample which prepare the biological treatment units in vegetable oils factories as active sludge, other bacterial types have been diagnosed by using traditional methods and API 20 E system. For detection of bacterial lipolytic activity, two methods were used for this purpose, the first was the screening of bacterial lipolytic activity which was based on values of clear zones diameter around bacterial colony. The best values were between (1.9 _ 2.7) cm. The second test was carried by the Gas - Chromatography - Flame ionization detector, fatty acids solutions that were produced from hydrolysis by lipase enzyme were extracted by Petrolium ether solvent then analyzed by gas chromatograph apparatus. The second type of test was the Molecular diagnosis by using the Polymerase Chain Reaction (PCR) to detect lipA gene in Pseudomonas aeruginosa by two primers. Through two primers used, lipA 948 was the best and more specialized primer to isolation lipA gene of P. aeruginosa. It gave (100%) a positive result. While the second primer lipA 558, gave (66.66%) a positive result and that may be due to the design of this oligonucleotides was not specific for lipA of P.aeruginosa. This is so as was based on highly preserved region of 12 bacterial lipA - homologous genes for many genus and many species belong to Pseudomonas. DNA sequencing done for amplicon generated using lipA 948, this sequence aligned by using BLASTn software against NCBI database to validate the results and investigate the similarity degree with other corresponding strains.
