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Detection the presence of Bdellovibrio bacteriovorus in the bacterial DNA extracts of Culex pipiens intestine using polymerase chain reaction

Ali Hassan Daghir Janabi

Al-Qadisiyah Journal of Veterinary Medicine Sciences مجلة القادسية لعلوم الطب البيطري
ISSN: 18185746 23134429 Year: 2017 Volume: 16 Issue: 2 Pages: 18-21
Publisher: Al-Qadisiyah University جامعة القادسية

Abstract

Mosquitoes are tiny creatures but could do a lot of damages to the people life and especially by transmitting dangerous diseases such as dengue fever transported by Aedes aegypti. The control of these bugs is hostorically challenging and difficult. This difficulty increases after the emerging problems of insecticide resistance. The gut flora(GF) in different animals has been detected to place an important role on the physiology of the intestine. This study was conducted to discover if the Bdellovibrio bacteriovorus is a member of the GF of the mosquitoes. B. bacteriovorus is a bacterium that has been found in the intestine of animals and healthy and patient humans but not in mosquitoes. Seventy adult mosquitos of Culex pipiens were used for extraction of the bacterial DNA from their intestine. The 70 mosquitoes were assigned randomly to 7 groups, 10 mosquitoes for each group to pool the amount of DNA extracted. Each mosquito was dissected under the microscope to isolate the intestine and use it later for DNA extraction. After the DNA had been extracted using a phenol/chloroform method, I subjected the extracted DNA to polymerase chain reaction (PCR) to detect the presence of this bacterium among the bacteria of the intestine using the following specific primers: Bd529F (5’-GGTAAGACGAGGGATCCT-3’) and Bd1007R (5’-TCTTCCAGTACATGTCAAG-3’) that amplify a 481-bp of the 16S rRNA gene. By the amplification that happened in all 7 groups, the result indicates the presence of Bdellovibrio bacteriovorus in the intestine of the Culex pipiens. Because this bacterium preys on Gram-negative bacteria, our results help to use this bacterium to fight insecticide resistance that caused by the degradation of these chemicals by gram-negative bacteria in the ingestion of these mosquitoes.