COMBINED MATING TECHNIQUE FOR ASSESSING CONJUGAL TRANSFER BETWEEN TEST VIBRIO CHOLERAE 01 MZW 41 AND PSEUDOMONAS AERUGINOSA W 109 + Wiaam Ahmed Al – Amili * Abstract This study presents results of comparing four in vitro conjugal transfer techniques, colony cross streak (CCS), broth mating (BM), combined spread plate (CSP), and membrane filtration (MF) with each other in evaluating the transfer of cojugative plasmids. Vibrio cholerae 01 MZW 41 and Pseudomonas aeruginosa W109, were used as mating pairs. The suitability of each method was evaluated for use in a routine assessment of the genetic stability of genetically transconjugated microorganism. By the BM technique, transconjugants were usually produced in 100% of the mating traits. In general, differences in the percentage of successful mating show that BM technique is statically significant at the P ≤ 0.05 level, compared with CCS, CSP, and MF techniques. Since the genetic exchange between Vibrio and Pseudomonas was not detected previously (an exception to this, what has been reported by Al – Amili, W.A. (2001), and any single conjugation transconjugants. We have used a combined mating technique which integrates the CCS, CSP, BM, and MF method as a single procedure to assess the mobility of plasmid DNA of genetically transconjugated microorganisms. ﺍﻝﻤﺴﺘﺨﻠﺹ ﺘﻅﻬﺭ ﻫﺫﻩ ﺍﻝﺩﺭﺍﺴﺔ ﻨﺘﺎﺌﺞ ﻤ ﻘﺎﺭﻨﺔ ﺃﺭﺒﻌﺔ ﺘﻘﻨﻴﺎﺕ ﺇﻨﺘﻘﺎل ﺇﻗﺘﺭﺍﻨﻲ ﻓﻲ ﺍﻝﺯﺠﺎﺝ ﻫﻲ، ﺍﻝﺘﺨﻁـﻴﻁ ﺍﻝﻤﺘﻘـﺎﻁﻊ ﻝﻠﻤﺴﺘﻌﻤﺭﺍﺕ) CCS ( ، ﺍﻝﺘﺯﺍﻭﺝ ﺒﺎﻝﺴﺎﺌل) BM ( ، ﻁﺒﻕ ﺍﻝﻨﺸﺭ ﺍﻝﻤﺭﻜﺏ) CSP ( ، ﻭﺍﻝﺘﺭﺸﻴﺢ ﺍﻝﻐـﺸﺎﺌﻲ) MF ( ﻤﻊ ﺒﻌﻀﻬﺎ ﻓﻲ ﺘﻘﻴﻴﻡ ﺇﻨﺘﻘﺎل ﺍﻝﺒﻼﺯﻤﻴﺩﺍﺕ ﺍﻻﻗﺘﺭﺍﻨﻴﺔ. ﺒﻜﺘﺭﻴﺎ ﻀﻤﺎﺕ ﺍﻝﻜﻭﻝﻴﺭﺍ01 MZW 41 ﻭ ﺒﻜﺘﺭﻴﺎ ﺍﻝﺯﺍﺌﻔـﺔ ﺍﻝﺯﻨﺠﺎﺭ ﻴﺔW109 ﺴﺘﺨﺩﻤﺕ ﻜﺄﺯﻭﺍﺝ ﺘﺯﺍﻭﺝ ﺃ. ﻤﻼﺌﻤﺔ ﻜل ﻁﺭﻴﻘﺔ ﻗﻴﻤﺕ ﻝﻼﺴﺘﺨﺩﺍﻡ ﻓﻲ ﺍﻝﺘﻘﺩﻴﺭ ﺍﻝﺭﻭﺘﻴﻨﻲ ﻝﻠﺜﺒﺎﺕ ﹰ ﺍﻝﻭﺭﺍﺜﻲ ﻝﻠﻤﻴﻜﺭﻭﺒﺎﺕ ﺍﻝﻤﻘﺘﺭﻨﺔ ﻭﺭﺍﺜﻴﺎ. ﺒﺈﺴﺘﺨﺩﺍﻡ ﺘﻘﻨﻴﺔBM ، ﺍﻨﺘﺠﺕ ﺍﻝﻤﻘﺘﺭﻨﺎﺕ ﻋﺎﺩﺓ ﻨﺴﺒﺔ100 % ﻝﺨﻭﺍﺹ ﺍﻝﺘـﺯﺍﻭﺝ، ﺒـﺼﻭﺭﺓ ﻋﺎﻤـﺔ. ﺍﻻﺨﺘﻼﻓﺎﺕ ﻓﻲ ﺍﻝﻨﺴﺒﺔ ﺍﻝﻤﺌﻭﻴﺔ ﻝﻠﺘﺯﺍﻭﺠﺎﺕ ﺍﻝﻨﺎﺠﺤﺔ ﺍﻅﻬﺭﺕ ﺍﻥ ﺘﻘﻨﻴﺔBM ﹰ ﻋﻨﺩ ﻤﺴﺘﻭﻯ ﻜﺎﻨﺕ ﻤﻌﻨﻭﻴﺔ ﺍﺤﺼﺎﺌﻴﺎ P ≤ 0.05 ﻤﻘﺎﺭﻨﺔ ﻤﻊ ﺘﻘﻨﻴﺎﺕCCS, CSP ﻭMF . ﺒﻤﺎ ﺍﻥ ﺍﻝﺘﺒﺎﺩل ﺍﻝﻭﺭﺍﺜﻲ ﺒﻴﻥ ﺒﻜﺘﺭﻴﺎ ﺍﻝﻀﻤﺎﺕ ﻭﺒﻜﺘﺭﻴـﺎ ﹰ ﺍﻝﺯﺍﺌﻔﺔ ﻝﻡ ﻴﻼﺤﻅ ﻤﺴﺒﻘﺎ) ﻋﺩﺍ ﻤﺎ ﻨﺸﺭ ﻤﻥ ﻗﺒلAL – Amili, W.A. (2001) .( ﻭ ﺇﻥ ﺍﻱ ﺘﻘﻨﻴـﺔ ﺘـﺯﺍﻭﺝ ﺍﻗﺘﺭﺍﻨﻲ ﻤﻔﺭﺩﺓ ﻻ ﺘﻜﻭﻥ ﻤﻭﺜﻭﻗﺔ ﺒﺼ ﻭﺭﺓ ﺘﺎﻤﺔ ﻓﻲ ﻤﻼﺤﻅﺔ ﺍﻝﻤﻘﺘﺭﻨﺎﺕ، ﻜﺒﺔ ﻝﻠﺘﺯﺍﻭﺝ ﻭﺍﻝﺘـﻲ ﺇﺴﺘﺨﺩﻤﻨﺎ ﺘﻘﻨﻴﺔ ﻤﺭ + Received on ٢ / ٤ / ٢٠٠٦ , Accepted on ٢٤ / ٧ / ٢٠٠٦ . * Asst Lectruter\Commission Of Technical Education College Of Medical Technologies/ Baghdad/ Department Of Medical Analysis