الفهرس 
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Abstract
Water is one of the main sources of life for all living organisms. Water Pollution could be a hazard due to infect transmission. Therefore, in our study, 60 pathogenic isolates were obtained from different sources of water used in our lives and identified the causes of pollution and recommending to reduce this pollution.
Plant extracts was used to inhibit the growth of pathogenic bacteria and a trial to replace drug antibiotics or reduce concentration of antibiotics used by mixture it with natural plant extracts to reduce the side effects of drugs and to change the effectiveness of antibiotic against the resistance of the microbe.
Aim of the study:
Isolation of pathogenic bacteria from different sources of water and studying the effect of some plant extracts on the water borne pathogenic bacteria . A trial to study the synergistic effect of some medicinal plant extracts with antibiotic drugs against water borne microorganisms with the lowest concentration of antibiotic.
The results obtained in this study can be summarized as follows:
1- Sixty water borne pathogenic bacteria isolates were obtained from 9 water samples from a total of 24 samples collected from 6 different sources namely Tap water (5 samples), Storage water (5samples) , Bottled water (5 samples), Underground water (2 samples), Renal dialysis water (5 samples) and Nile water (2 samples) which were collected from four governorates ( Cairo, Giza, Sharqia and Qalyubia).The Nile water was recorded as the highest level of contaminate being 45.7% (Shubra El-Mazalat area in Qalyubia governorate and El-Warraq area in Giza governorate ) followed by two samples of Tap water being 28.6% (Embaba in Giza and Belbeis in Sharqia) then 2 samples for Storage water (2 hospitals in Giza and Cairo) and Underground water being 11.42% (2 houses of Ain Shams in Cairo and Belbeis in Sharqia) then one sample of Renal dialysis water being 2.86% (hospital in Cairo ) , whereas the Bottled water was not recorded for any bacterial count . The contaminated samples for Storage water , Renal dialysis water and Underground water (one sample from house of Ain Shams in Cairo)were free from Total coliforms, Fecal coliforms and Fecal streptococci. The twenty four samples were all free from Salmonella sp. , Shigella sp., Vibrio sp. and Staph. aureus . Therefore it could be stated that Giza governorate had the highest density of water borne pathogenic bacteria followed by Sharqia and Cairo being 41,33,22 and 4 % respectively.
2- Three chemical tests were carried and to determine the chemical characteristics of water (Turbidity, pH and Residual chlorine). Results show that all chemical tests were in the normal rate except one sample from renal dialysis water which recorded high pH reached 8.2 whereas the pH normal rate for renal dialysis water was 6.8 –7.4 .
3- Sixty isolates were identified using morphological specifications, biochemical tests and serological tests by antisera for Escherichia coli and Enterococcus groups. All the tested Escherichia coli (isolates) were not related either to E.coli O175 or E.coli O175: H 7) , Whereas antisera for Enterococcus groups revealed that they were Enterococcus group D (Enterococcus faecalis).
4- Among sixty bacterial isolates, thirty five isolates were identified based on the molecular weight of the ribosome proteins by MALDI-TOF-MS. Ninteen isolates were identify as Enterobacter spp. (14 isolates were Enterobacter cloacae, 3 isolates were Enterobacter asburiae, one isolate of Enterobacter kobei and one isolate of Enterobacter hormaechei), 5 isolates Escherichia coli, 4 isolates Enterococcus faecalis, 3 isolates Klebsiella pneumonia, 2 isolates Citrobacter freundii, 2 isolates Pseudomonas aeruginosa. Enterobacter cloacae for the rest of the thesis tests.
5- The isolate number 19 which isolated from renal dialysis water in specialized hospital in Cairo was identified as Enterobacter cloacae DSM 3264 BRB with high confidence score (score value) being 2.17 , whereas isolate No.37 which isolated from underground water (Sharqia) was identified as Enterobacter cloacae MB11506_1 CHB with score value 2.32 . Enterobacter cloacae DSM 3264 BRB will taken for further studies due to their high risky for renal failure patients.
6- The antibiotic sensitivity test was carried out at different concentrations for each antibiotic starting from the in optimum concentration until the least concentration 0.25 µg/100µl. The zone of inhibition by Gentamicin against Enterobacter cloacae DSM3264 BRB was ranged between 1.05 – 2.2 cm with average 1.517 cm, Ciprofloxacin 1.4 – 2.6 cm with average 2.070 cm, Meropenem 1.55-2.850 cm with average 2.217 cm and Cefepime (CPM) were 1.25 – 2.9 cm with average 2.056 cm. It’s interesting to note, that the recommended dose of the tested antibiotics were 10, 5, 10 and 30 µg/100µl respectively. Comparing the obtained data with standard zone of inhibition against Gram negative bacilli (Enterobacteriace) using standard concentrations of different antibiotics according to CLSI (2015). Data revealed that the tested strain was susceptible (S) to antibiotics Gentamicin, Ciprofloxacin, Meropenem and Cefepime within range 2-10, 1-5, 2-10 and 10-30 µgl/100µl respectively. Otherwise, it was resistant (R) to the previous antibiotics concentration within range 0.25-0.50 , 0.25, 0.25-0.50 , 0.25-1.00 µg/ 100µl respectively.
7- The results showed that the minimal inhibitory concentration (MIC) of the bacteria using antibiotics ranged from 0.25 to 5 µg/100µl. Gentamicin had the lowest for MIC at 0.25 µg/100µl, Ciprofloxcin at 1.0 µg/100µl and then Meropenem and Cefepime at 5 µg/100µl.
8- The tested strain was susceptible to Cinnamon aqueous and methanol as well as Clove methanolic extracts, but the highly effective was Clove oil extract being 2.6 cm as mean diameters of inhibition zone . All oil plant extracts revealed that the highest inhibition zones ranged from 1.3 to 2.6 cm except Black seed and Basil which didn’t gave any inhibiting activity . Aqueous water and methanolic extracts showed the lowest inhibition zone against E.cloacae DSM 3264 BRB expressed as less than 1.35 cm.
9- The minimal inhibitory concentration of Cinnamon oil, Cloves and Mint was recorded at 0.5, 5.0 and 5.0 μl / 100 µl respectively, so Cinnamon oil was more effective in bacteria.
10- Minimal bactericidal concentration (MBC) was determined for Cinnamon, Clove and Mint oil , it was found that the MBC values of the tested plants against Enterobacter cloacae DSM 3264 BRB were 0.5, 5.0, 5.0 (µl/100µl) respectively . The tested plant oils extracts considered as bactericidal affect against Enterobacter cloacae DSM 3264 BRB. Since , MBC/MIC= 1.0 .
11- Mixture design was used to determine the option concentration of plant extract using 20 treatment trials. The significant treatment was the fourth, which recorded Gentamicin 0.25% (w / v) + 5% of Cloves oil + 5% of Mint oil + 4.75% (v/v) of Cinnamon oil where it gave an inhibition of 2.7 cm diameter.
12- The results of the Gas chromatographic analysis were determined using the quantitative analysis of hydro distilled Essential oils of the three selected plant oils (Mint , Clove and Cinnamon) using GC mass. results that Mint oil contains is composed of 27 chemical compounds which it were 4 compounds with an effective effect against the bacteria representing 66.08% of the essential Mint oil (menthol is the largest components with ratio of 32.92% followed by menthone 27.75%, pulegone 4.06% and m-Mentha-1,8-diene (1.3528%)) , while the essential oil of Cinnamon contains 3 compounds antibacterial effect representing 84.93% (cinnamic acid, phenethyl ester is found to have the highest component of (41.97%) and followed by cinnamaldehyde (24.42%) and 3-Allyl-6-methoxyphenol(m-Eugenol) (18.55%). while the essential oil of Clove compounds antibacterial effect (3-Allyl-6-methoxyphenol(m-Eugenol)) representing 46.41%.
Recommendations :
• Using MALDI-TOF-MS in the identification of pathogenic bacteria .
• The results of this study show the possibility of reducing the concentration of antibiotic to 40 times with the use of a combination of medical oils using the program of mixing design used in the pharmacy to determine the best quantities of each element to reach the best inhibitory effect on the bacteria.
• We recommended to study their effect within the body of the living organism in addition to the side effects of the preparation and also the possibility of using plant extracts in the diets to protect them from rapid microbial corruption and prolong the life of the food and also increase the daily use in our lives (food, drinks and external use of the skin) for beneficial effect and it protects against bacteriological diseases and other diseases.
• The study also revealed that there are areas of pollution in drinking tap water due to the lack of maintenance and renovation of pipes and also in the storage water due to lack of periodic cleanliness and recommended not to use the underground water before analysis and also, the obligation to analyze the water of the dialysis after preparation and before it is used for patients and reduce the dumping of drainage water Nile to reduce the spread of pollution and diseases