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Diagnosis of honeybee diseases using PCR and laser spectroscopic techniques /

Amal Mohamed Ibrahim Osman

Diagnosis of honeybee diseases using PCR and laser spectroscopic techniques / و استطياف الليزر PCRتشخيص امراض نحل العسل باستخدام تقنيات ال Amal Mohamed Ibrahim Osman ; Supervised Mona Abdelaziz Mohamed , Emtithal Mohammed Abdelsamiea , Hamdy Taher Mohamed Abouelenain - Cairo : Amal Mohamed Ibrahim Osman , 2015 - 152 P. : charts , photographs ; 25cm

Thesis (M.Sc.) - Cairo University - National Institute of Laser Enhanced Science - Department of Laser Application in Environmental Metrology Photochemistry and Agriculture

Paenibacillus larvae subsp. larvae, the causative pathogenic bacterium of virulent brood disease (American foulbrood, AFB). DNA extracted from P. l. larvae strain was subjected to PCR using specific primers of a partial sequence of 16S rRNA gene that closely related to the bacterium. Assayed bacterial strain produced a defined band of 700 bp in size. This test was done based on DNA extraction of bacterial colonies and different larval stages(1-5 days, of age) from Carniolan (Apis mellifera carnica) honeybee and Italian (Apis mellifera ligustica), The results of PCR analysis showed that the (day1) of all honeybee races didn't amplify the pathogen fragment (700bp) and all larval stages from (day2- day5) generated PCR fragment of 700bp. Also, this defined fragment was detected in larvae, pupae, adult worker and virgin queen resulted from the grafting Italian honeybee. On the other hand, the physicochemical structure of genomic DNA of pathogenic bacterium P. l. larvae and the two honeybee races before (-ve control) &after infection with the pathogen were identified and characterized qualitatively and quantitatively using molecular laser Raman (MLRS) and infra-red (FTIR) spectroscopies for the first time. The results of spectroscopic analysis clearly revealed that the pathogen was more abundant in Italian race from the early infection stage (day1) to (day5) of age. than Carniolan which showed more resistant to pathogen infection of the (day2)



Carniolan and Italian honeybee races P. l. larvae PCR