الفهرس 
LITERATURE REVIEWOnly 14 pages are availabe for public view
 |  | from | 222 |  |  |
| | | from | 222 | | |
Abstract
The present study was conducted to evaluate the potential role of Culex pipiens adult female mosquitoes in the transmission of hepatitis b and c virus (HBV& HCV), after fed on an infected blood through feeding membrane apparatus with a viral load of 1.9 x 106 IU/ml in HBV experiments and 1.2 x 106 IU /ml in HCV experiments symbiotic and aposymbiotic mosquitoes.
Adult female mosquitoes head region, mid-gut and salivary glands at different intervals were tested for the detection of viral load using real-time PCR.
In case of HBV experimental infection, DNAs of HBVs detected for 5& 7 hours in head region and up to 4 & 5 days mid-gut of symbiotic and aposymbiotic mosquitoes respectively.
While in HCV experimental infection, RNAs of HCV detected for to 60 and 90 min in head region and up to 5 & 6 days in mid-gut of symbiotic and aposymbiotic groups respectively
Absence of HBV DNA and HCV RNA in salivary glands after 3, 6, 11, 12 and 13 days post experimental infection.
The rearing of the mosquito, Culex pipiens, was carried out at the insectary of Entomology Department, Faculty of Science, Ain Shams University, Cairo - Egypt.
All experimental infection was carried out at Virology Department, Faculty of Medicine, Al azhar University under controlled laboratory conditions at temperature of 272C, relative humidity 7010% and 12-12 light-dark regime.
HBV DNA and HCV RNA extraction and real time PCR analysis was carried out at VACSERA, Holding Company for Production of Vaccines, Sera and Drugs, Giza-Egypt.
Isolation and identification of gut bacterial microbiota were carried out in National Cancer Institute, Cairo - Egypt .
Results obtained can be summarized as follows:
1. Evaluation of blood feeding membrane technique.
Cattle blood with chick membrane in mosquito rearing experimental infection, vectorial capacity and pathogen interactions studies is a good substitute for human blood and live feeding for its availability and cost effectiveness.
2. Experimental infection with HBV in symbiotic and aposymbiotic adult female mosquitoes.
2.1. Evaluation of the viral load at head region of HBV symbiotic and aposymbiotic adult female mosquitoes:
Data obtained revealed that, there was a decrease in the percentage of the HBV titers by increasing the time, post feeding in symbiotic and aposymbiotic mosquitoes.
A negative correlation between one female HBV titers and time was observed.
The results indicated that the mechanical transmission of HBV through head region of symbiotic mosquitoes can be possible.
2.2. Evaluation of the viral load in mid-gut of HBV experimental infected adult female symbiotic and apo symbiotic mosquitoes:
Results showed that there was a decrease in the percentage of the HBV titers which induced by increasing the time with higher time and titers in aposymbiotic mosquitoes.
2.3. Evaluation of the viral load in salivary glands of HBV experimental infected adult female symbiotic and aposymbiotic mosquitoes:
Data given showed that, no detection of HBV DNA at 5, 6, 11, 12, and 13 days post blood feeding.
The results also indicated that the biological transmission of HBV through adult female symbiotic and aposymbiotic mosquitoes did not occur.
3. Experimental infection with HCV in symbiotic and aposymbiotic adult female mosquitoes.
3.1. Evaluation of the viral load in head region of HCV experimental infected adult female symbiotic and aposymbiotic mosquitoes:
Data given showed that, there was a decrease in the percentage of HCV titers by increasing the time. A negative correlation between female HCV titers and time
The results also indicated that the mechanical transmission through symbiotic and aposymbiotic mosquitoes head region may be possible.
3.2. Evaluation of the viral load in mid-gut of HCV experimental infected adult female symbiotic and aposymbiotic mosquitoes:
Results showed that there was a decrease in the percentage of HCV titers by increasing the time.
3.3. Evaluation of the viral load in salivary glands of HCV experimental infected adult fema symbiotic and aposymbiotic mosquitoes:
Data given showed that, no detection of HCV RNA at 5, 6, 11, 12, and 13 days post blood feeding.
The results also showed that the biological transmissions of HCV through symbiotic and aposymbiotic mosquitoes did not occur.
4. Experimental transmission of HBV and HCV by mosquitoes:
Results showed that, mosquitoes which harbored the virus and allowed to complete its diet directly after infection (with contaminated head region) could transmit HBV particles to HBV negative blood sample 2 (with load 69 x 104 IU/ml). While, mosquitoes which harboured the virus 2,4, 6 and 10 days post feeding could not transmit it at any time.
Mosquitoes which harbor HCV and allowed to complete its diet directly after infection (with contaminated head region) can transmit HCV particles to HCV – negative blood sample 2 (with load 78x104 IU/ml). While, mosquitoes which harbour the virus eight days after feeding could not transmit HCV particles.
5. Estimation of blood feeding, ovioposition, hatachability rates, survival and digestion period through HBV and HCV experimental infected adult female symbiotic and aposymbiotic mosquitoes.
5.1. Estimation of some biological paramters (blood feeding, ovioposition and hatachability rates) in HBV and HCV experimental infected symbiotic and aposymbiotic mosquitoes.
Data obtained showed that, blood feeding, ovioposition and hatachability rates decreased significantly among symbiotic and aposymbiotic mosquitoes in both HBV and HCV experimental infected symbiotic and aposymbiotic mosquitoes adult female mosquitoes.
Aposymbiotic mosquitoes showed greater decreasing than symbiotic mosquitoes.
5.2. Estimation the survival rate through HBV and HCV experimental infected adult female symbiotic and aposymbiotic mosquitoes:
As shown from the results, the survival rate decreased significantly in HBV and HCV infected aposymbiotic adult female mosquitoes more than HBV and HCV infected symbiotic mosquitoes by increasing the time post feeding .
5.3. Estimation of the digestion period through HBV and HCV experimental infected adult female symbiotic and aposymbiotic mosquitoes.
The results indicated that a very highly significant increase in the blood meal digestion period of HBV and HCV infected in aposymbiotic adult females and highly significant increase in the blood meal digestion period of HBV and HCV infected symbiotic females.
6. Isolation and characterizeration of some bacterial gut microbiota associated with Cx. pipiens.
Escherichia coli, Klebsiella ornithiolytica, Enterobacter cloacae, Acinetobacter triffolii and Acinetobacter baumannii are isolated and identified from gut of adult female mosquitoes as Gram positive strains members.
Staphylococcus aureus and Micrbacterium olevians are isolated and identified as Gram negative strains members.