الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
This study was conducted using 150 oral swap samples of dental plaque patients from the Oral medicine and periodontitis department – Faculty of Dentistry – Alexandria University. After purification and identification by: Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) biotype. Only 39 non contaminated and well identified bacterial isolates was selected. Two different honeybee strains, Apis mellifera yamentica, and Apis mellifera carnica from a farm in the Ganaklis area at Nobarya, Behaira governorate were used in this study. Bee venom was spectroscopically analyzed by HPLC to estimate the exact content of melittin and phospholipase A2 in every sample. The disc diffusion method was used to test the effect of bee venom on various bacterial isolates (with a mean of inhibition zones of 3 mm). Results proved that Gram-positive bacteria were more sensitive even at lower concentrations tested from Bee Venom than Gram negative bacteria. The studied bacterial isolates were resistant to one of the main components (Phospholipase A2). Wheras melitinin showed antimicrobial activity against many of the tested isolate with the highest antimicrobial activity recorded against Streptococcus mutans and Streptococcus mitis at a concentration of 50 µg/mL. The non-protein fraction has no inhibitory effect on the bacterial growth in all the tested bacterial isolates. Interestingly, the tested protein fraction had a moderate inhibitory effect on bacterial growth. The minimum inhibitory concentrations (MIC) of BV were determined with values ranged between 31.3 and 44.4 μg/mL, while the mean minimum bactericidal concentration (MBC) was nearly 30 μg/mL for all samples. Biofilms (dense micro-communities that grow on inert surfaces encapsulating themselves with secreted polymers). Photos were captured by an analytical Scanning Microscope at the Central Laboratory of City of Scientific Research & Applied Technology. Examination of the captured images showed an effect of different concentrations from bee venom on biofilm formation. Bee Venom sub-MICs totally inhibited Staphylococcus sciuri and candida albicans biofilm at concentrations of 25 and 20 μL of BV respectively.
Real Time PCR to showed Streptococcal collagen-like protein-1 (Scl-1) gene and Elongation factor-1 beta (EFB1) gene before and after treatment with bee venom. Data gained were expressed in Cycle threshold (Ct) for the target genes and the reference gene (16-S for bacteria and 18-S for Candida). Target gene expression was assessed and related to reference gene as Δ Ct = Ct assessed gene – Ct reference gene and the gene expression consequently could be detected from the (2 ^- Δ Δ Ct) value.
Gene expression levels for Streptococcal collagen-like protein-1 (Scl-1) gene in the five studied groups (ATCC standards, positive controls, group1, group 2, and group3) showed a decrease in expression in the bee venom-treated groups (with concentrations of 50 µL and 150 µL of bee venom) by (0.7; 0.7 and 0.8) and (0.7, 0.8 and 0.9). On the other hand, ATCC criteria, positive control and group 1 had increased expression in: ATCC standards (1.5, 1.5 and 1.4), positive control group (1.1, 1.0 and 0.9) and group 1 (1.1, 1.1 and 1.1) times, respectively. The gene expression levels in the untreated negative control group (ATCC criteria) were equal to (1) in the data analysis and calculation. This means that the SCl-1 gene was down-regulated in the bee venom-treated groups and was independent of dose.
On the other hand gene expression levels of Elongation factor-1 beta (EFB1) gene in different studied groups: ATCC standards, positive controls, group1, group 2, and group3 showed a decrease in expression in the groups treated with bee venom (with concentrations of: 50 µL and 150 µL of bee venom) per (0.8, 0.9 and 0.7) and (0.9, 1.0 and 0.8) folds. The ATCC criteria, the control was positive, and group 1 had an increased expression according to the ATCC criteria as follows (1.3, 1.3 and 1.2), the control group was positive as follows (1.4, 1.1 and 1.3) and group 1 was as follows (1.2, 1.1 and 1.1) times, respectively. The gene expression levels in the untreated negative control group (ATCC criteria) were equal to (1) in the data analysis and calculation. This means that the EFB-1 gene was down-regulated in the bee venom-treated groups and was independent of the dose.
Bee venom may be an effective complementary antimicrobial agent for use against oral pathogenic bacteria. Moreover, Bee venom showed a significantly more reduced effect on antibiotic-resistant bacteria in biofilms than on plankton or stalkless bacteria. Mellitin and phospholipase A2 contents of bee venom are affected by many factors like the season of extraction, time of collection, method of extraction, and the pollen grains that the bee eats. Phospholipase A2 has no antibacterial effect but, mellitin alone has an antibacterial effect more than bee venom as a whole. The presence of C. albicans in initial in vitro oral biofilms changes the community composition significantly. The streptococcus collagen-like protein-1 (Scl-1) and elongation factor beta (EFB1) genes in different groups were studied; showed a decrease in expression in the bee venom-treated groups (with 50 µl and 150 µl of bee venom). This means that the SCl-1 and EFB1 genes were down-regulated in the bee venom-treated groups and were dose-independent.