الفهرس 
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Abstract
The present study aims to study the evaluation of classical and molecular techniques in characterization and enumeration of coliform bacteria in water, also to study characterization of pathogenic bacteria and its resistance to different antibacterial agents. River Nile as a main source of drinking water in Egypt has become a matter of interest and one of the most important national goals. Damietta branch has a great importance since it is one of River Nile branches and the main source of water in the Delta region. Unfortunately, it receives a variety of wastes coming from sewage, agricultural and industrial drainage that affect its water quality. On the light of the previously mentioned objective, Ten water samples along Omar Bek drain and from Damietta branch of Nile River, (1st January – 31 December 2016; from 8.00 a.m. to 10.00 a.m. - air temperature was 31oC), to study and show the influence of different pollution sources on the Nile River. These samples were subjected to physico-chemical and microbiological analysis. The results of the study can be summarized as follows: Eco-diversity of water of Omar bek drain included Physical and chemical factors as following: 1- Temperature values of water samples ranged from 23.4°C to 25.8°C with note the absence of any source of thermal pollution. 2- pH values for all collected water samples are slightly higher (7.62-9.23) than the permissible limits of law 48/1982. 3- EC values for water samples collected from Omar bek drain were recorded elevated, where they fluctuated between (0.43 – 2.25 dS/cm) 4- TDS concentrations for water samples collected from Omar bek drain are exceeds than the permissible limits of law 48/1982 (< 500 mg/l) except for the point from Damietta branch before the location of Omar Bek drain (468.3 mg/l ) and point from Damietta branch after Omar Bek drain mixing point with the branch (480.5 mg/l). 5- Ammonia concentrations for water samples collected from Omar bek drain not exceeded the permissible limits of the law 48/1982 (<0.5 mg/l), where it ranged from (1.02 mg/l to 1.94 mg/l,). 6- DO values for water samples collected from Rosetta branch were within the permissible limits of law 48/1982 (<5 mg/l) where they fluctuated between (0.2 – 5.56 mg/l). 7- BOD concentrations for water samples collected from Omar bek drain were higher than the permissible limits in the law 48/1982 (> 10 mg/l) and ranged between (2 – 50 mg/l). Except for the point from Damietta branch before the location of Omar Bek drain and point from Damietta branch after Omar Bek drain mixing point with the branch were recorded (3 & 1 mg/l, respectively) and within the permissible limits. 8- COD concentrations for water samples collected from Omar bek drain were variable compared with the permissible limits in the law 48/1982 (>15 mg/l) and ranged between (5 -73mg/l). Eco-diversity of Soil located in Omar bek drain area included Physical and chemical factors as following: 1- Organic matter percentage values of soil samples beside Omar Bek drain were ranged from (1.37% – 5.25%) with mean of 3.61%. 2- pH value of soil samples was ranged from (7.20 – 8.24) and slightly higher than the normal range (6.5 – 8.5). 3- The recorded values of Electrical Conductivity (EC) and Total Dissolved Solids (TDS) were within the range of the permissible values of law 48 / 1982 limits for drain water; Carbonates ions were not detected (0.00) in all sites 4- Saturation percentage for soil samples collected from Omar bek drain were ranged from (85-90 %) for all sites along the studied area. 5- The dominant anions mean can be arranged in the following descending order Cl-> SO4 -2> NO3 - > PO4 -3 respectively. On the other hand, Na+ represented the dominant cation followed by Ca2+ and Mg2+ whereas K+ means values were the lowest one. The quantitative biodiversity for microorganisms was done in according to the following methods: 1- Standard plate count (SPC) bacteria at 22°C and 37°C in all collected water and soil samples recorded high values and varied regionally. The highest value was recorded at 3 points (A meeting point between three other drains from Omar Bek drain; A point after sewage treatment plant from the drain and A drain that discharges into Omar Bek drain). 2- Bacterial indicators (total coliform, fecal coliform and fecal streptococci) in all collected water samples exceeded the international permissible limits. To testify our potential method, E. coli was used as the control bacteria for it. After RNA extraction, serial dilutions were made to check our ability to extract a standard curve and equation using these dilutions, 16s RNA as the reference gene, lacZ genetargeting primers and probes and qPCR for the quantification. Total coliform counts for the 10 sampling locations using membrane filtration technique and qPCR method were compared. from results, it is obvious that qRT-PCR method counts for total coliform was so near to the results obtained by MFT method by 102.4±14%. Identification of bacteria involved isolation of “455” isolates, out of which “375” and “80” isolates were identified into “19” different species. About 180 pathogenic isolates were identified from both soil and water samples. The biodiversity of isolated bacteria from soil and water using different antibiotics. The antibiotic sensitivity test was performed using 17 commercially prepared antibiotic discs. The identified bacterial isolates (19) either from soil or water of Omar bek drain showed multiple antibiotic resistances against the individual antibiotics used with different percentages. Resistance was 100% against, Ampicillin (AM10) and Oxacillin (OX1). The results of antibiotics resistance patterns for different pathogenic bacterial isolates showed that E. coli isolates were resistant to about (75%), C. freundii isolates (50%), S. enteric isolates (80%), S. liquefaciens isolates (60%), P. vulgaris isolates (75%), P. aeruginosa isolates (75%), E. faecalis isolates (35%), M. catarrhalis isolates (65%), K. pneumonia isolates (50%), Y. pestis isolates (97%), E. aerogenes isolates (45%), S. sonnei isolates (65%), E. cacticidia isolates (50%), P. stuartii isolates (45%), H. alveiisolates (65%), A. hydrophila isolates (35%), and S. aureus isolates (70%) of all tested antibiotics. The antibacterial activity of nature antibacterial solvents (organic solvents) and biosynthesized silver nano-particles were performed against both gram positive bacteria (S. aureus and E. faecalis) and gram negative bacteria (P. aeruginosa, S. liquefaciens, E. aerogenes, K. pneumonia, S. sonnei, L. pneumophila, Y. pestis, M. catarrhalis and H. alvei) by well diffusion method. The minimum inhibition of growth diameter obtained with the acetone extract of S. platensis against bacterial pathogens was comparatively very less when compared to the other solvent extracts used in this study. On the other hand, there was no inhibition of growth diameter was seen with DMSO solvent. The silver nano-particles synthesized from S. platensis solution showed an effective antibacterial activity against all of tested bacteria strains at volume of 200 μl/well. S. platensis has been studied because of its therapeutic properties and the presence of bioactive compounds. But nano material + water + DMSO were not effective enough against some of tested inhibition of growth diameters of the pathogenic bacteria as follow: S. aureus, S. liquefaciens, L. pneumophila, Y. pestis, M. catarrhalis and H. alvei. The lowest inhibition of growth diameter with nano-particle solution synthesized by S. platensis may due to the aggregation of silver nano-particles that synthesized by S. platensis as shown in results For example and as a result, the nature phytochemical solvents synthesized from S. platensis solution was able on inhibiting of growth diameter of the H. alvei 5X3 isolate more than nano material synthesized from the S. platensis. The nature antibacterial reagents and nano material were synthesized from S. platensis solution were able to on inhibiting of growth diameter of the pathogenic bacteria more than antibiotics used in this study.