الفهرس 
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Abstract
The present work aimed to investigate the efficacy of some natural essential oils on the microbial vaginitis especially bacterial and yeast vaginitis as a new promising alternative remedy. In this study fifty clinical microbial vaginitis were isolated from ten different groups each contains numbers of cases. These groups were arranged as the following; normal healthy, microbial vaginosis, pre and post-vaginal area, pregnant, nosocomial infection, unexplained infertility, hyperglycemia with obesity, child with poor hygiene, dysfunctions of the immune system and parasitic vaginal infection cases. The obtained data showed 4 bacterial isolates were isolated from 4 cases in the first group without fungal or parasitic pathogens, from the second groups; 11 bacterial and 2 fungal isolates were recognized with no parasites. The third group showed 2 bacterial isolates were isolated from 2 cases and no fungal and parasitic isolates. from 12 cases of the fourth group, 10 bacterial and 2 fungal isolates were isolated and no parasitic isolates. Six bacterial isolates were isolated from 6 cases of the fifth group. from 2 cases of the sixth group 2 bacterial were isolated and no fungal and no parasitical isolates. Four bacterial and three fungal isolates were isolated and no parasitic isolates from 7 cases in the seventh group. from the eighth group, one parasitical isolate was isolated from 1 case. The ninth group showed 2 bacterial isolates were isolated from 2 cases without fungal or parasitical pathogens. The tenth group which contains one case showed no isolated bacterial and fungal isolates but only 1 parasitic isolate was isolated. ”Amsel’s” diagnostic criteria were used for detection and differential diagnosis of the isolated microbial vaginitis. These criteria (diagnostic parameters) were pH, clue cells, whiff test and vaginal discharge nature. All the isolated microbial isolates were cultured using different selective media and were examined morphologically, microscopically using a magnifying lens observation for the shape of colony, margin, elevation, surface (texture), and chromogenesis. Also each of the isolated microbial vaginitis were identified using fully automated VITEK2 Compact system, 47 isolates were identified as following: 20 Klebsiella pneumoniae, 4 Staphylococcus aureus, 6 Escherichia coli, 2 Pseudomonas aeruginosa, 5 Lactobacillus plantarum, 1 Neisseria gonorrhoeae, 1 Gardnerella vaginalis, 1 Acinetobacter baumannii, 6 Candida albicans and 1 Candida lusitaniae. One bacterium isolate of Peptostreptococcus prevotii was identified using ID32A rapid kit identification test strip for anaerobic bacteria. 2 parasitical isolates were microscopically identified, 1 isolate of Enterobius vermicularis eggs, and 1 isolate of Trichomonas vaginalis trophozoites. The antibiotics sensitivity of the tested bacterial isolates showed different susceptibilities ranging from sensitive (S), intermediate (I) and resistant (R) reactions against sixteen tested antibiotics. The results showed that out of the thirty seven pathogenic bacterial isolates, nine bacterial isolates exhibited multidrug resistance against the used antibiotics, theses bacterial isolates were L. plantarum, A. baumannii, N. gonorrhoeae, G. vaginalis, E. coli, P. aeruginosa, K. pneumoniae, S. aureus and P. prevotii where the inhibition zones ranged between 6 to 10 millimeter (mm) in diameters. While the yeast isolates showed different susceptibilities reactions against the five tested antibiotics. Two isolates out of the seven isolates of yeasts, C. albicans and C. lusitaniae showed MDR against 4 and 3 of the used antibiotics respectively. Seventeen antimicrobial plant’s essential oils were tested against the nine MDR bacterial and the two MDR yeast. The results illustrated that turmeric, garlic, black seed and parsley oils exhibited the highest ntibacterial and antifungal activities against the MDR bacteria and yeast isolates. where the recorded inhibition zones were 38, 32, 21, 10 mm for the K. pneumoniae isolate no. 28, and 33, 27, 11, 10 mm for the C. albicans isolate no. 49, respectively. The identification of K. pneumoniae and C. albicans by VITEK2 compact system was confirmed by molecular identification of 16s rRNA gene sequence using PCR for the bacterial isolates. The results showed that the selected bacterial isolate exhibited a similarity of 98% to the 16s ribosomal RNA sequence of Klebsiella pneumoniae subsp pneumoniae at accession no. OP02045. And for identification of C. albicans by 18s rDNA the comparison gave 98% similarity to C. albicans at accession no. OP023836. The effect of combination of the selected resisted antibiotics with each of the most potent essential oils, turmeric, garlic, black seed and parsley oils against the selected pathogenic MDR K. pneumoniae and C. albicans showed that the combination of the antibiotic levofloxacin with each of the essential oils turmeric, garlic and black seed had slightly increasing in the inhibitory activity against the MDR K. pneumoniae followed by high inhibitory effect with using parsley oil. While the combination of the antibiotic nystatin with turmeric oil had the highest inhibitory effect against C. albicans where the diameter of inhibition zone increased from 33 to 40mm. This result exhibited that the effect of combination of nystatin antibiotic with turmeric oil recorded the highest inhibitory effect against MDR C. albicans that is might be breaking down its resistant to antibiotic nystatin. The biofilm formation by the selected K. pneumoniae and C. albicans were carried out and the results illustrated that the formation of biofilm by the two isolates were recorded. The effect of turmeric oil at different concentrations (100, 50, 25, 12.5, 6.25, 3.125, 1.562, 0.781, 0.390, 0.195 and 0.097) on the dry weight and biofilm formation for both K. pneumoniae and C. albicans was studied. The results showed that increasing in the turmeric oil concentrations caused a decrease in both dry weight and biofilm formation for each the selected MDR K. pneumoniae and C. albicans, compared with non-treated isolates which produce high amounts of biofilm in the absence of turmeric oil. These results indicated that the biofilm formation of both isolates were decreased with increasing turmeric oil concentrations, so this oil can be used as anti-virulence factor against the two isolates K. pneumoniae and C. albicans. The effect of turmeric oil at concentration 0.781 mg/ml on the biofilm formation by K. pneumoniae and C. albicans at intervals times (time course) was carried out to determine the time required for the highest inhibition of the biofilm formation of the two selected MDR isolates. The results showed that the time course required for maximum inhibition of biofilm formation by the selected MDR K. pneumoniae and C. albicans were recorded 2.2 and 3.5 μg/ml at 8 hours compared with untreated control 8 and 10 μg/ml respectively, this indicated that the treatment of MDR microbial vaginitis with turmeric oil caused great inhibition to the biofilm formation and break down the resistance of microbial vaginitis against antibiotics at 8 hours. Microbicidal and microbistatic effect of turmeric oil at concentration 0.781 mg/ml on the growth of K. pneumoniae and C. albicans in liquid media at different time intervals were studied to determine the time-kills kinetics assay. The results showed that, with increasing time from 0 to 12, 24, 36, 48, 60 then 72 hours, the optical densities (OD) of untreated K. pneumonia were increased gradually with increasing the time from 0.215 at zero time to 0.712 at 72 hours. As well as OD for untreated C. albicans was increased from 0.165 at zero time to 0.635 at 72 h. The effect of turmeric oil on the OD of K. pneumoniae was increased gradually from 0.215 at zero time to reach 0.308 at 72 h. While the effect of turmeric oil on OD of C. albicans were increased from 0.165 to 0.211 at 72h. On the other hand, the CFU for untreated K. pneumoniae control growth was increased from 0.170 to 0.730 at 72 h. As well as untreated C. albicans control CFU were increased gradually from 0.120 to 0.680. Oil treated K. pneumonia CFU were increased to 0.170, 0.190, 0.210, 0.220, 0.230 at 48h, then decreased to 0.00, and 0.00 at 60 and 70 hour respectively, while treated C. albicans CFU were gradually increased from 0.120 to 0.141at 48h., then decreased to 0.00 at 60 and 72 hour respectively. These reported values indicated that turmeric oil had a cidal effect on the tested microbes at 60 and 72 hours. Scanning Electron Microscopy was conducted to observe the morphological changes that occurred to K. pneumoniae and C. albicans after treatment with turmeric oil at 0.781 concentration for the two isolates. The scanned images showed severe physical damage to the tested microbes and lead to irregular cell shapes with the destroyed cell wall and shrinking of cells. Some of the cells were vacant, while other cells were flimsy. Additionally, most of them appeared to be melted and jammed together. K. pneumoniae cells had enormous appendages on their surface. While treated C. albicans showed structures resembling pseudo mycelium. The untreated control cells of K. pneumonia and C. albicans were whole and had smooth surfaces. GC-MS analysis was performed to detect the active antimicrobial materials of turmeric oil. GC-MS determined the percentages and structure of the major components, revealing Aromatic-turmerone as a major ingredient at 58.033%. These and other components in the analyzed oil were expected to be responsible for the previously recorded antimicrobial activity of turmeric oil against vaginal pathogens.