Vol. 48, February, 2015

1/48 ISOLATION AND IDENTIFICATION OF PRODUCING EXOPOLYSACCHARIDE EGYPTIAN ISOLATE (ENTEROBACTER COWANII, 26)

Bahgat M. Refaat, Yasser R. Abdel-Fattah*, Nadia A. Soliman*, Mohamed Helal El-Sayed and Mahmoud K. Abdel-Fattah.

Botany and Microbiology Department, Faculty of Science (Boys); Al-Azhar University, Nasr City, Cairo, Egypt.

Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications (SRTA City), Universities and Research Institutes Zone, New Borg El-Arab, Alexandria, Egypt.

Exopolysaccharides (EPS) are environment friendly natural polymers secreted by microorganisms in the surrounding medium. Due to the presence of unique structural composition, EPS shows diverse applications such as food formulations, pharmaceutical, and cement based construction industry, etc. This study aimed at the isolation of bacteria producing high yield of exopolysaccharide among the bacteria isolated from plant samples showing characteristics of infection with target bacteria. The most potent bacterial isolate has been identified through culture characteristics, biochemical and finally at the molecular level using 16S rDNA. The sequence alignment with data base at the National Center for Biotechnology Information NCBI revealed that this isolate has maximum similarity with Enterobacter cowaniiHQ407245 with an identity 99%. This isolate has been submitted to the GeneBank under accession number KP070767. After production, extraction and purification, exopolysaccharides have been characterized using physical, chemical and spectrophotometric analysis. The result indicated that; exopolysaccharide has aldehyde or keto groups and the monomeric composition contains glucose, rhamnose. The purified exopolysaccharide showed fine emulsification and viscosity properties that required further application including toxicity studies, pharmacological testing, and in-vivo application in animal model in order to fulfill the long term goal using of this polysaccharide at different industrial and pharmacological application.

2/48 OPTIMIZATION OF FERMENTATION CONDITIONS FOR BIOETHANOL PRODUCTION BY SACCHAROMYCES CEREVISIAE ON EGYPTIAN BEET MOLASSESUSING TAGUCHI ORTHOGONAL ARRAY DESIGN.

Mohsen A. Sayed, Tahany M. Abd-Elrahman, Islam M. Hamdy

Department of Microbiology -Faculty of Science- Cairo University
Bioethanol has attracting growing interest around the world in order to reduce greenhouse gas emission and reduce the dependence on petroleum fuels. The present work focused on the enhancement of ethanol productivity and yield in the fermentation system using sugar beet (Beta vulgaris) molasses as substrate for S. cerevisiae fermentation activity. Optimization of the cultural conditions for maximum production of ethanol was performed using the taguchi statistical design with investigating eight factors and 18 experimental media. After ANOVA analysis, it was found that inoculum size was the only significant factor for sugar consumption, while biotin, pH, and specific gravity were significant factors in bioethanol production. Ethanol production from beet molasses by S. cerevisiae under the optimized conditions revealed that medium 16 was the optimum for sugar consumption (100 %), fermentation efficiency (99.61), degree of attenuation (62%), bioethanol yield (Y p/s =0.508 g/g ), bioethanol productivity (Q p =1.966 g/500ml/h) and remaining sugar (0.00 g/500ml).

3/48 PHYTOCHEMICAL AND TAXONOMICAL STUDIES ON SOME UMBELLIFEROUS PLANTS (APIACEAE)

Mamdouh M. Zayed, Mohamed K.M. Hamza,El-sayed H. El-Kafafi,

Ahmed B.I. Khalil, El-Sayed A.E. Abd El-Aziz

Botany Department, Faculty of Agriculture, Azhar University.

The analysis of seed volatile oils in eight species of Apiaceae showed the presence of 46 different components in studied samples. The results showed also that there were specific components in each sample except Pimpinella anisum. These special compounds may be used to recognize these plant samples. Results of the analysis of seeds volatile oils were used to calculate the similarity coefficient between these plant samples. The largest similarity was between Apium garveolens and Petroselinum sativum, while the lowest similarity was between Anethum garveolens and Foeniculum vulgare.


4/48 MICROBIAL BIOSYNTHESIS OF POLYHYDROXYBUTERATE FROM ANAEROBIC DEGRADATION PRODUCTS.

Essam A.S. Raguab, Mohamed H.M. Abd el-Megid and Hussein M. El- Kabbany*

Natural Product Radiation Research Dept., *Health Radiation Research Dept.

National Center for Radiation Research and Technology, Atomic Energy Authority, Cairo, Egypt.

Three anearobic bacterial strains have been shown to accumulate glycogen during active growth; Rhodopseudomonus capsulata RC 15 (the wild type) and the two mutant strains of Rh capsulata (RC 15), (R 25 and C 5), were cultivated in glutamate-maleate (GM) medium and screened for production of polyhydroxybutyrate (PHB). The mutant strains R25 and C5 were found to accumulate PHB (53.9 and 42.0% of dry cell weight (DCW), respectively), 3.6 and 2.8 times higher than the wild type strain (19.5% of DCW), respectively. R. capsulataR15 were selected the effective of nutrient and environmental conditions on PHB accumulation. The optimal condition was 4 g/l acetate, 0.02 g/l (NH4)2SO4, C/N ratio of 6:1, 1.0 g/l K2HPO4, 1.0 g/l KH2PO4 and 3% Na Cl with initial pH at 7.0. Under this optimal condition, the maximum PHB accumulation increased from 53.9% to 88% of DCW and 9.11 ± 0.08 g/l biomass, 8.02 ± 0.10 g/l PHB concentration were achieved after 60 hrs cultivation at 37ºC. This study revealed that this effluent can be used as substrate for PHB (bioplastic) production. These can be degraded by microorganisms without any chemicals. So we can conclude that bio-plastic, an eco-friendly, a very valuable product and fungi isolates degrade the PHB (bio-plastic) much higher than that of bacteria.

5/48 ASSOCIATION OF PEGYLATED INTERFERON ALPHA / RIBAVIRIN-INDUCED PITUITARY AND ADRENAL HORMONES DYSFUNCTION IN PATIENTS WITH CHRONIC HEPATITIS C VIRUS INFECTION[S1]

Shaden Muawia,Amal A. Abdelaziz*, Ghada M. Nasr**, Amany Gamal*** and Sohir E. Shoman****

Molecular Biology Department, (GEBRI), Sadat City University, Sadat City, Egypt

*Molecular Biology Department, Dean of (GEBRI), Sadat City University, Sadat City, Egypt

**Molecular Diagnostics Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), Sadat City University, Sadat City, Egypt

***Biochemistry Department, National Hepatology and Tropical Medicine Research Institute, Cairo, Egypt

****Tropical Medicine Department, National Hepatology and Tropical Medicine Research Institute, Cairo, Egypt

Chronic hepatitis C virus (HCV) infection is one of the most widely spread diseases in the world and highly prevalent in Egypt. Chronic infection with HCV can result in both hepatic and extra hepatic diseases. Endocrine dysfunction represents an important class of HCV-related extra hepatic disease.To determine the association of pegylated interferon alpha (IFN)and ribavirin-induced pituitary and adrenal hormones dysfunction with severity of disease and response to treatment in patients of chronic hepatitis C infection. We have investigated pituitary hormone secretions in 70 patients with chronic hepatitis C virus infection, before and after12 weeksof antiviral therapy (pegylated interferon-α plus ribavirin). These included: Growth hormone (GH), cortisol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), thyroid-stimulating hormone (TSH), testosterone (free and total), sex hormones, and dehydroepiandrosterone sulfate (DHEAS). Growth hormone concentration was increased significantly during therapy. Basal PRL and TSH concentrations were normal before and after therapy. Cortisol, and testosterone concentrations were also normal. Investigated sex hormones and DHEAS levels in chronic viral hepatitis were not decreased. Neither the HCV RNA levels nor transaminases were correlated with hormone’s concentrations before or after therapy.

6/48 EFFECT OF BACTERIAL INDICATOR POLLUTION ON SURVIVAL GROWTH OF AQUATIC AND FREE LIVING AMOEBAE.

Hussein M. El- Kabbany

Health Radiation Research Dept., National Center for Radiation Research and Technology, Atomic Energy Authority.

The recently observed high levels of fecal indicators in beach sand can found beach water monitoring efforts. The high levels of fecal indicators may be caused by the loss or the reduced activities of common environmental stresses controlling die-off in the sand. Experiments were conducted to compare the effects of biotic stresses from sand bacteria, protozoa, and viruses on Escherichia coli and Staphylococcus faecalis in two tropical beach sands. The inhibition of protozoan activities by cycloheximide did not significantly affect the die-off of E. coli, indicating that protozoan predate on played a limited role in beach sand. The contribution from phage infection to E. coli die-off was also negligible. Consequently, bacteria were identified as the predominant biotic stress to the die-off of E. coli in beach sand. Subsequent experiments demonstrated that the beach sand had a very low protozoan concentrated and low protozoan growth potential when compared with various environmental samples, the study also found that E. faecalis exhibited a much longer survival in beach sand compared with E. coli.

7/48 INTEGRATED BIOTIC AND ABIOTIC AGENTS APPLICATION IS A NECESSITY TO IMPROVE BARELY PRODUCTION IN SALT-STRESSED ENVIRONMENTS                                   

Hassan Ismail, Manal Abd Allahand Heba Shehata

Soils, Water and Environ. Inst., ARC.

Salinity is one of the dangerous factors affecting the growth and production of plant. It makes unbalance in nutrient uptake, disorder in physiological operations, decrease in height of shoot and root; reduce in leaf area and many symptoms that indicate plant unhealthy. In the present study, the addition of ascorbic acid and/or extracts of either Spirulina platensis or bio-straw inoculated with Phanerochaete chrysosporium supported the plant ability to withstand the salinity, increased the grain yield and improved the metabolic activity. The mixture of ascorbic acid and extracts of both Spirulina platensis and bio-straw was the auspicious treatment that raised the grain yield to 2.53 ton/fed. comparing to 1.26 ton/fed in control. The mixture of abiotic and biotic agents increased the chlorophyll (a) content that reached to 22.4 mg/g plant compared to 12.9 mg/g plant in control. This points out the plant health, encourages excretion of proline that has an important role as osmo-protectant, enhances the performance of dehydrogenase activity that denotes the viability of microflora in rhizosphere region and increase the N P K uptake.

8/48 ISOLATION AND IDENTIFICATION OF STAPHYLOCOCCUS SPP. ISOLATED FROM ORAL SPECIMENS IN EGYPTIAN PATIENT

Saad A.M. Moghannem, Mohamed H. Elsayed,Bahgat M. Refaat,

Gamal M. Elsherbiny, Hussein Abo El-Fetouh Abd El-Fadeel and

Elsayed Abd El-Kreem Mahna*

Plant & Microbiology Department, Faculty of Science (Boy’s), Al-Azhar University, Nasr City, Cairo,

*Dentist at Faculty of Dentistry (Boy’s), Al-Azhar University, Nasr City, Cairo, Egypt.

This study reports the isolation and identification of Staphylococcus spp. by using BIOMÉRIEUX VITEK®2 SYSTEM in clinical specimens and Antibiotic Susceptibility Profile as well as determination of some virulence factors from different Egyptian clinics. 208 clinical specimens were collected from different patients including wound, pulpitis, plaque, pulpectomy, tartar, ulcer, damage molar, abscess, diabetic patients. The isolation was performed using specific culture media for selection and confirmed biochemically. Virulence factors are important in identification of bacterial invading tools which cause pathogenesis that can be used as new targets in drug development. A total of 52 Staphylococcus spp. were isolated. From these staphylococci, it was found that 9 isolates (17%) were highly antibiotic resistant, 22 (42.4%) coagulase positive (CoPS), 30 (57.6%) coagulase negative (CoNS), 8 (15.4%) have the ability to make Alpha (α) hemolysis on blood agar, 23 (44.2%) have Beta (β) hemolysis and 21 (40.4%) have Gamma (γ) hemolysis activity. From 22 (42.4%) coagulase positive (CoPS), 4 isolates (18.2%) exhibits resistance to Oxacillin while 6 isolates (27.2%) were intermediate and 12 (54.6%) were sensitive. In addition to, Vancomycin resistant strains were 2 (9.1%) isolates, while 5 (22.7%) were intermediate and 15 (68.2%) were sensitive.

9/48 EFFICIENCY OF SALICYLIC ACID AND GLYCINE ON SUGAR BEET PLANTS GROWN UNDER HEAVY METALS POLLUTION

Emad A. Ewais, Mohamed A. Ismail and Mohamed A. Amin and Ali A. Badawy

Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, Egypt.

A pot experiment was conducted to evaluate the beneficial effect of foliar application of salicylic acid (50 & 100 ppm) and glycine (50 & 100 ppm) on growth and some biochemical constituents in plants grown in soil polluted with cadmium (Cd) or lead (Pb). Heavy metals caused significant decreases in most of the growth, photosynthetic pigments and yield characteristic of sugar beet plants. Treatment with salicylic acid (SA) or glycine (Gly) improved plant growth and yield as well as some metabolic contents in plants. The changes in endogenous hormones, proteins and carbohydrates contents were interrelated. Cd or Pb addition to soil increased their contents in both shoot and root of sugar beet plants whereas; SA or Gly reduced the uptake and accumulation of heavy metals in the tested plants.

10/48 INCIDENCE OF ENTEROTOXIGENIC STAPHYLOCOCCI AND YEASTS IN RAS CHEESE LAYERS

Talat Ibrahim EL-Sayed

Botany Department, Faculty of Science, Benha University

Fourty-five samples of Ras cheese layers were collected from supermarkets, representing the surface, the middle and the core layers of Ras cheese. The mean counts values of Staphylococcus aureus from each layer were 4.544 ± 4.342, 2.230 ± 1.602 and 0respectively; only, two enterotoxigenic strains could produce enterotoxin type A, while, the mean values of yeasts were 4.462 ± 3.204, 2.255 ± 0.380 and 2.322 ± 0.919. The most common isolated yeast species were Candida, Rhodotorula, Torulopsis and Saccharomyces. These results indicate that the core layers of Ras cheese were more safe microbiologically than the surface layers. The chemical examination of Ras cheese showed that pH, acidity % and moisture % were 4.30 ± 0.100, 1.62 ±0.033 and 32.3 ±0.014, respectively. The public health of examined Ras cheese and the control measures to safeguard the consumers were discussed.

11/48 BIOGENESIS OF RADIOLYTIC SILVER NANOPARTICLES via Aspergillus oryzae And THEIR ANTIMICROBIAL IMPACT

Eman Ahmed Mohamed Helmy

The Regional Center for Mycology and Biotechnology (RCMB), Al-Azhar University, Cairo, Egypt.

Radiolytic nanoparticle manufacturing granted an economical and environmentally-safely attempts to make metallic nanoparticles in significant with slim dimension dispersity. Aqueous supernatant of Aspergillus oryzae was operated as a securely tool for the establishment of long-standing silver nanoparticles (AgNPs) by way of gamma irradiation at doses from 1 to 30 kGy. The volume and the amount of the produced silver nanoparticles (AgNPs) was gamma dose-dependent as it could be altered by managing the radiation dose. The AgNPs outcomes were verified using UV-Visible spectrophotometry, dynamic light scattering (DLS), Fourier Transform infra red (FT-IR) spectrophotometry, X-ray diffraction (XRD), Energy Dispersive X- ray (EDX) and transmission electron microscope (TEM) analysis which displayed the ultra-morphology of globular AgNPs with 10–17 nm in diameter. The synthesized AgNPs revealed antimicrobial activity against both Gram positive and Gram negative bacteria, as well as some pathogenic filamentous fungi and yeasts as evaluated by well diffusion assay technique. In the present study a nanofactory system to synthesis rapid, eco-friendly and narrow size range of AgNPs was introduced.

12/48 AMELIORATIVE EFFECT OF BIOFERTILIZERS ON THE OXIDATIVE STRESS AND IONIC COMPOSITION OF COWPEA UNDER SALINITY STRESS

Ibrahim M. Zeid, Safia M. Ghazi, Nermin A. El Semary and Amira M. Abd El-Sattar

Botany Department, Faculty of Science, Helwan University, Cairo, Egypt

The present study was carried out to investigate the response of cowpea plant grown under saline conditions to two types of microorganisms as biofertilizers e.g., Aphanizomenon sp. (cyanobacteria) and Mycorrhiza (fungi). Salinity stress stimulated the activity of oxidases and antioxidant enzymes e.g., (ascorbic acid oxidase, polyphenol oxidase, IAA oxidase, catalase and peroxidase) and non enzymatic antioxidant system e.g., (phenolic and flavnoids compounds). It also reduced the mineral ions content. The lipid-peroxidation and relative permeability of the root membranes increased as compared with control. Application of aqueous extract of the cyanobacterium Aphanizomenon sp. and/or the mycorrhiza treatment increased some ions content of leaves, as well as the K+/Na+ ratio. Lipid-peroxidation and relative permeability of root membranes were reduced in salt-stressed and unstressed plants due to the treatment with biofertilizers. So, the application of these microorganisms as biofertilizers enhanced the plant resistance to salinity stress may be through improving the water status and the cellular metabolism. The biofertiliers can be used as a cost-effective, eco-friendly farming approach to counteract the hazardous effect of salinity on plants.

13/48 BIODEGRADATION OF 4-CHLOROPHENOL BY SOME BACTERIAL ISOLATES FROM EGYPTIAN PETROLEUM OIL SPILLS

Mamdouh.S.El-Gamal, *Mervat .A.M.Abo-State, Essam.H.Abd El-Shakour and Abdallah.M. Abdo

Department of Botany and Microbiology, Faculty of Science, Al-Azhar University

* National Center for Radiation Research and Technology (NCRRT)

4-Chlorophenol (4-CP), a major type of chlorophenols, has been widely used to produce some herbicides and pharmaceuticals, yet due to its incomplete degradation and bioaccumulation characteristics, it is very toxic to living organisms. So, Microbial biodegradation is a promising technology to clean up environments contaminated with that recalcitrant compound.In this study, we investigate biodegradation of 4-Chlorophenol (4-CP) by five indigenous bacterial isolates from Egyptian petroleum oil wastes. These isolates were grown on five concentrations (500, 1000, 2000, 3000 & 4000μM) of 4-chlorophenol (4-CP) as a sole carbon and energy source .The ability of each isolate to grow and degrade 4-CP have been determined by recording the growth (O.D) (nm), Secretion of extracellular protein (µg/ml) and Cl- ions release (mg/L) at (initial) time and 1, 2, 3, 7, 14, 21 days incubation. Also the cell counts (CFU/ml) were determined initially (zero time) & after 21 days incubation period. Also The abilities of these chlorophenols degrading bacterial isolates to degrade 4-CP have been determined quantitatively after 21 days using High Performance Liquid Chromatography (HPLC).

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