Dr. Osama Hassan Mansour El-Sayed :: Publications:

This study aimed to either examine the stability of transferred foreign Deoxyribonucleic acid (DNA) in chicken seminal plasma or to investigate the effect of introducing broiler hybrid chicken-DNA into embryonic cells of Mamourah chicken using spermatozoa as a vectors which known as sperm-mediated gene transfer technique on fertility and hatchability percentages as well as embryonic mortality of Mamourah incubated egg. Trial I: A band of 709-pb extracted from pEGFP-C1 plasmid DNA after cutting by BsrGI and AgeI restriction enzymes used as DNA substrate to evaluate the DNAse activity in chicken plasma semen of Mamourah chicken. Volume of 10µl fragment-DNA substrate was incubated at 40C for 40 min within four different tube contain distilled water, distilled water+ chicken seminal plasma, distilled water+ chicken seminal plasma+ EDTA and distilled water+ chicken seminal plasma+ Lipofectin reagent. Then, DNA substrate was detected by electrophoresis on 1% agarose gel containing ethidium bromide and the DNA stability was detected under UV light. Obtained results concluded that the DNA substrate was completely diminished after incubation with seminal plasma diluted in distilled water. However, the DNA substrate was stabilized after incubation with chicken seminal plasma in the presence of either EDTA or lipofectin reagent. Trail II: A total number of 150 matured Mamourah pullets aged 8 months were randomly grouped into six groups (25 pullets per each group). Pullets of the first three groups were artificially inseminated with semen of the same strain containing spermatozoa used as vectors for transferring level 20, 25 and 30µg of broiler hybrid-DNA (male) diluted with skim milk media in the presence of lipofectin reagent and considered as treated groups. Pullets of the other three groups were artificially inseminated with semen diluted with either media in the presence of Lipofectin reagent, semen diluted with only media or untreated semen and considered as control groups. It was clearly found that applying foreign DNA level significantly decreased the fertility percentage of eggs collected from treated pullets. The rate of decrement increased as the level of foreign DNA applied increased. The lowest average of fertility percentage was detected in pullets of the first generation mounted 71.85%. However, it increased in birds of the second generation to reach an average mounted 90.37%. On the other hand, no significant variation in the average of hatchability percentage due to treatments applied. Moreover, hatchability percentage did not show any significant variation due to pullet's generation. However, hatchability percent was slightly higher (88.93%) in pullets of the first generation when compared to those of pullets of the second generation (86.85%).