Vol. 20, May, 2008

1/20 tumor-inhibitory l-asperaginase: production by different fungal isolates and partial purification and characterization from penicillium oxalicum

Sahar M. Ouda

Plant Research Department, Nuclear Research Center, Atomic Energy Authority, Cairo, Egypt.

Seven fungal isolates under the present study were able to produce L-aspraginase in different quantities in presence of L-asparagine. Optimal substrate concentration for enzyme production was 1% for P .oxalicum, T. harzianum and A. terreus after 5 days incubation period, while it was at 0.2% for A. niger, T. longibrachiatum, A. flavus and S. cerevisiae after 3 days incubation for A. niger and T. longibrachiatum while after 5 days for A. flavus & S. cerevisiae. P. oxalicum showed the highest extracellular L-asparaginase production followed by T. harzianum, A. terreus and A. niger (264.5, 257.3, 223.9 & 177.9 mg/L), respectively. 0.2% L-proline maximize the production for P. oxalicum, it reached 306.7 mg/L after 5 days incubation, while 0.2% glutamine increased the production of A. terreus after 3 days incubation (266.6 mg/L). Glucose was the best C-source comparing with sucrose and manitol. Ammonium sulphate at 40-60% saturation recorded (4.44 fold) specific activity, partial purification of precipitated enzyme on sephadex G-100 showed activity peak at fractions no. 7, 8 & 9 & specific activity was 41.85 mMole / mg protein / ml with purification fold 5.41. Characterization of partially purified asparginase was carried out, the results were found to be as follows: optimal incubation temperature was 50°C, thermal stability up to 50°C, incubation time 15 min., substrate concentration 20 uMole & pH of acetate buffer was 7.

dapoxetine en france 2/20 ANTIFUNGAL ACTIVITY OF SOME SOIL ALGAE AGAINST GROWTH AND PHYSIOLOGICAL ACTIVITIES OF SOIL FUNGI

Nemmat A. Hussein

Botany Department, Faculty of Science, Assiut University, Egypt

The inhibitory effect of different concentrations (5%, 10% and 15%) of extracts of three algal species ( priligy en france 2016 Chlorella fusca var. cialis et maladie cardiaque vaculata, Microsystis aeruginosa and forum meilleur site achat cialis Oscillatoria angustissima) isolated from soil have been investigated against soil fungi. The results of the primary screening indicated that generique tadalafil 20mg prix Microsystis aeruginosa inhibited the growth of most fungal species, and this inhibitory effect increase by increasing algal concentrations. On the other hand. The fungal total counts increased by addition of douleur testicule cialis Chlorella fusca var. temps effet kamagra vaculata and cialis rythme cardiaque Oscillatoria angustissima to soil, although the numbers of genera and species reduced compared with control. Many fungi ( il cialis altera le analisi del sangue Acremonium aegyptiacum, Acrophialophora fusispora, Aspergillus terreus var priligy en pharmacie en france aureus, A. versicolor, Cochliobolus lunatus, C. spicifer, Paecilomyces lilacinus, Penicillium duclauxii, P. pinophilum, Rhizopus stolonifer var . lyoccocus and Stachybotrys chartarum) were disappeared in the presence of algal suspension. Mucor racemosus is the only fungal species that was either slightly stimulated or unaffected by the three algal species tested. Oscillatoria angustissima showed the most inhibitory effect on fungal dry weight. The metabolic response (carbohydrates, amino acids and proteins content) and enzyme activity were determined in three fungal species (Aspergillus niger, Fusarium solani and Mucor racemosus). Carbohydrates and amino acids content of tested fungi were significantly reduced by the action of different concentrations of algal suspension added caused decrease in peroxidase activity of Aspergillus niger. Finally, Chlorella showed no inhibitory effect on fungal growth and metabolic activity, this may be due to the ability of fungi to secret antifungal toxins against Chlorella.

3/20 DECOLORIZATION OF DYES BY MICROALGAE

Hanan H. Omar

Botany Dept., Faculty of Science, Tanta University, Tanta, Egypt.

In the present study, 11 isolates of green algae, cyanobacteria and diatom were evaluated for their ability to decolorize wide range structurally different synthetic dyes. All isolates decolorized some dyes, and however some dyes were decolorized to some extent. The maximum decolorization was attained with the lower concentration of dyes. The most color loss was observed within the first 3 days after which, the rates of removal slowed up to 6 days. The percentage decolorization varied from 100% to only 13% after 6 days incubation. Alizarine color was completely disappeared within 24h, whereas the maximum decolorization was found to be 90 to 96 % with Methyl violet, Methylene Blue and Alizarine Red in algal cultures. The algal cultures exhibited the minimum dye removal with Eosine (31 to 77%) and Acid Green (13 to 46%). Among the tested algae, S. bijugatus (green algae) and N. muscourm (cyanobacteria) were most effective for dye removal. The color removal efficiency could be related to the molecular structure of individual dyes and the species of algae used. Equilibrium isotherms were well described by the Langmuir equation, giving maximum affinity binding of dyes with green algae > cyanobacteria > diatom. The IR spectra of algae were carried out before and after treatment with dyes to identify the functional groups of algae which adsorb the dye molecules. The results indicate that the algae can be used as an excellent low-cost adsorbent for dyes.

4/20 SUPPRESSION OF SOIL-BORNE PATHOGENS OF SUGARBEET PLANT BY CHITOSAN DERIVED FROM CARB-SHELL CHITIN IN VIVO AND IN VITRO

Nahid Z. Haikal, Dalia M.I. Ali and Mohamed Eweis

Department of Botany, Faculty of Science, University of Cairo, Giza Egypt.

The antifungal behavior of chitosan, derived from crab-shell chitin, was studied in in vivo and in vitro. Chitosan applied to soil at (100, 200, 300 and 400 µg g-1 soil) suppressed infection of two sugarbeet cultivars kawmera and Trivave by the fungal pathogens Rhizoctonia solani kühn (AG2-2) and Sclerotium rolfsii Sacc. and acting thus as a fungicide. Virulence was directly correlated with the activities of cell wall degrading enzymes produced by these pathogens in vivo. Reduced virulence of R. solani and S. rolfsii under chitosan stress was due to decrease inoculum potential of the two pathogens at all utilized concentrations of the fungicide in situ and to reduce production of cell wall degrading enzymes in vivo. In vitro studied, the purified chitosan had a significant inhibiting effect upon the growth activities of the tested sugarbeet pathgens R. solani and S. rolfsii in the polymer concentration range of (100, 200, 300 and 400) µg ml-1/ medium. The rates of cell wall degradation of kawmera and Trirave by the in vitro produced R. solani enzymes were nearly 2.5 and 5 times, respectively, faster than by S. rolfsii enzymes produced in the absence of chitosan. Under chitosan stress, the cell wall degrading activity of culture supernatants, of either pathogen decreased with increasing chitosan concentration. However, the activity of R. solani supernatants was consistently higher than those of S. rolfsii in all treatments. The pH of the cell wall amended culture media of the two cultivars shifted towards alkalinity in R. solani and acidity in S. rolfsii. The result was confirmed by the SDS-PAGE for cellular proteins of both R. solani and S. rolfsii, which showed a great variation in protein bands for the same fungus at different chitosan concentrations.

5/20 ENHANCING REMOVAL OF MERCURY FROM HYDROPONIC PLANT GROWTH BY MERCURY-RESISTANT PLASMID PNC7 FROM BACILLUS SUBTILIS NC

Mohamed E.A. Dawoud and Youssef A. Mawgoud

Botany Department, Faculty of Science, Cairo University, Giza Egypt.

The mercuric reductase of Bacillus subtilis NC was proved to be plasmid-borne genes. The plasmid pNC7 was purified and partially characterized with some antibiotics (gentamycin and sulphonamide) and some restriction enzymes sites (BamHI, EcoRI, PstI, and SAu3A). Vicia faba L. and Zea mays L. seedlings were grown in mercury-hydroponic solution, fortified with B. subtilis NC (harboring plasmid pNC7). Growth criteria of tested plants (% plant length, % Dry weight and % chlorophyll leaf content), and the binding of mercury to the growing bacterial cells, indicated sharply the capability of pNC7 plasmid to cope with mercury contamination in the rhizosphere niche area.

6/20 OPTIMIZATION OF PHYTASE ENZYME PRODUCTION FROM HALOTOLERANT ASPERGILLUS NIGER AND ITS USE IN PROMOTION OFWHEAT GROWTH UNDER SALT STRESS.

Mohamed A. Foaad

Department of Biological and Geological Sciences. Faculty of Education, Ain Shams University, Heliopolis, Roxy, Cairo, Egypt.

The reduction of phytate content in plant feeds is advisable for increasing their nutritional values. Phytase improves the bioavailability of phytate phosphorous in plant foods and reduces phosphorous pollution of animal wastes. In the present work halotolerant strain of Aspergillus niger was isolated from soil of Qarun lake coast and investigated to produce phytase enzyme. The isolate was used to produce extracellular phytase by using wheat bran as sole phosphorus source. Optimization of the culture conditions for maximum production of the enzyme was investigated .High activity of enzyme (162.6+1.23U/ml) occurred on PSM medium supplemented with wheat bran 10; glucose 40; ammonium nitrate 2.5 (g / l) at pH6 and 30oC. The partially purified enzyme has two optimum pH peaks at 5 and 7.5. and maximal activity at 55oC. The activity of the enzyme was reduced by heat treatment and 98.7 % of its activity was lost at 70 o C .The culture supernatant of the fungus at maximum phytase activity as well as the spores of the fungus have promoted wheat seedling growth under salt stress and increased its phosphorus content.  


7/20 APPLICATION OF PLACKETT-BURMAN EXPERIMENTAL DESIGN TO EVALUATE FACTORS AFFECTING ANTAGONISTIC EFFECT OF BACILLUS SP. AGAINST THE PLANT PATHOGEN FUSARIUM OXYSPORUM F.SP. LYCOPERSICI

Mona E.M. Mabrouk and *Ebaa Ebrahim El-Sharouny

Botany Department, Faculty of Science, Damanhour Branch, Alexandria University, Egypt

*Botany Department, Faculty of Science, Alexandria University, Egypt.

Twenty bacterial strains were tested for their potentiality to produce bioactive compounds that inhibit the growth of the plant pathogen F. oxysporum f.sp. lycopersici. Bacillus sp. produced a compound that showed highest activity against the tested organism. Temperature 35°C and pH 7 were found to be the best for maximum antifungal production. High level of glucose and yeast extract and low levels of K2HPO4 in the culture medium had the most signnificant effect on the production of the antagonistic compound as evaluated by the application of Plackett- Burman experimental design. Antagonistic activity of the compound produced from the optimized culture conditions against F. oxysporum f.sp. lycopersici, showed 20 % increase than that obtained from the basal medium. Cell-free culture ļ¬ltrates of Bacillus sp. inhibited conidial germination and hyphal growth of F. oxysporum f.sp. lycopersici. The inhibitory substance was found to be affected by protinase K and high temperature.

8/20 EFFECT OF SOME PLANT EXTRACTS AND VOLATILE OILS FOR CONTROLLING SUGARBEET ROOT-ROT DISEASE CAUSED BY SCLEROTIUM ROLFSII SACC. IN VIVO

AND IN VITRO

Nahid Z. Haikal, Ahmed M. Emam* and Dalia M.I. Ali

Botany Department, Faculty of Science, Cairo University, Giza, Egypt.

*Biochemistry Department, Faculty of Agriculture, Fayoum University,

Fayoum, Egypt

The goal of this study was designed to evaluate the possible effects of different plant extracts as well as commercial volatile purified oils for controlling sugarbeet root-rot disease caused by Sclerotium rolfsii Sacc. Ethanolic-acetone extracts (1:1 v/v) from two different plants represent seeds of: Ammi visnaga L. (pick tooth), Trigonella foenum-graecum L. (fenugreek). as well as two commercial purified volatile oils of Syzygium aromaticum L. (clove) and Cuminum cyminum L. (cumin), were screened for their antifungal activity in vitro and in vivo. In vitro, the effect of different concentrations (500, 1000, 1500 and 2000 µg m-1) of ethyl alcohol and acetone (1:1 v/v) plant seeds extracts as well as purified volatile oils were tested on the growth activities of Sclerotium rolfsii, the causal pathogen of sugarbeet root-rot disease, on Czapek's-Dox medium. Both of seeds and volatile oils had a significant inhibiting effect on the different stages of development of the fungus including the percent of germination of sclerotia, mycelial growth and the number of sclerotia produced. The inhibitory effect was proportional with the applied concentration as well as the nature of bioactive compounds that found in the plants. In vivo, growth parameters including emergence, plant length and dry weight of sugarbeet cultivar kawmera were significantly increased at all utilized seed extracts as well as purified volatile oil concentrations (500, 1000, 1500 and 2000 µg g-1 soil). The infection rate was significantly decreased with increasing concentrations, whereas the total reducing value (TRV) and the direct reducing value (DRV) were significantly increased in the tested sugarbeet cultivar kawmera.

9/20 EVALUATION OF SPICES AS ANTIMICROBIAL COMPOUNDS  

Mohsen Abou El-Ela, Tahany Abdel-Rahman and Nagwa Tharwat

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

The antimicrobial susceptibility of fifteen spice plants was evaluated against four fungal species: Aspergillus niger, A. flavus, Fusarium solani and Penicillium chrysogenum; and four bacterial species: two Gram positive (Bacillus subtilis and Staphylococcus aureus) and two Gram negative (Escherichia coli and Proteus vulgaris). Clove completely inhibited the growth of the four tested fungi and significantly inhibited bacterial growth. Significant inhibitory effect was also observed in case of cinnamon and garlic, meanwhile, mint, licorice, carob tree, fenugreek, henna, african doum and rose mallow moderately inhibited tested microorganisms. Other tested spice plants showed a low inhibitory effect on tested microbes. Mixing of ethanolic extracts of two spices alter their antimicrobial activity such that mixing of two spices with no antimicrobial activity may result in creating antimicrobial activity when used together. Aspergillus niger was the most susceptible fungus to the inhibitory effect of spices. Gram negative bacteria were more resistant than Gram positive bacteria to the inhibitory effect of the tested spices.

10/20 RESPONSE OF ACID PHOSPHATASE ACTIVITY IN THE HOUSE FLY MUSCA DOMESTICA (DIPTERA: MUSCIDAE) TO SOME INSECT GROWTH REGULATORS

Abdel-Hamed A. Abdel-Ghaffar, Mohamed A. Tanani and Karim S. Ghoneim

Faculty of Science, Al-Azhar University, Madenit Nasr, Cairo, Egypt

After feeding the early last instar larvae of Musca domestica on Lufenuron-treated diet (10.0, 1.0 and 0.1 ppm), an increasing trend of ACP activity was detected along the pupal stage with an exception of those pupae after larval treatment with the highest concentration of Lufenuron since the enzyme activity was depressed in the last pupal day (1.51±0.021 vs 1.69±0.007 U/ml of control congeners). However, Lufenuron failed to inhibit the ACP activity in statistically significant value at beginning of the pupal stage (at all concentrations). Irrespective of the given concentration level (0.1, 0.01 or 0.001 ppm) of Diofenolan through the larval diet, ACP activity increased during the first two pupal days and then decreased in the 2- and 3-day old pupae but unexpectedly resumed to increase at the end of the stage (1.58±0.011, 1.65±0.012 and 1.67±0.018 U/ml, at 0.1, 0.01 and 0.001 ppm, respectively, vs 1.69±0.007 U/L of control congeners). However, Diofenolan exhibited a considerable inhibitory effect on ACP activity in the pupae of all ages, at the higher two concentration levels. After topical application of Lufenuron and Diofenolan (0.1, 0.01 and 0.001 µg/larva of both IGRs) onto late last instar larvae, ACP activity during the pupal stage was gradually inhibited with age but appeared in an increment at the end. Also, the 1-day old pupae had promoted ACP activity by the action of Lufenuron. A significant reducing effect of Diofenolan was detected with no exception.

11/20 PRELIMINARY STUDIES CONCERNING THE MICROBIOLOGICAL QUALITY OF SOME EGYPTIAN POPULAR FOODS SOLD IN CENTRAL AREAS OF TANTA CITY.

Mohamed S. Salem, Mostafa Aboul-Fadl, Ashraf M. Sharaf

and Sameha M. El-Saied*

Food Sci. & Technol. Depart., Faculty of Agric., Al-Azhar Univ. Cairo.

*Food Sci. & Technol. Depart., Faculty of Home Econ., Al-Azhar Univ.Tanta.

A quantitative survey issued the basic information of microbial magnitude for 130 samples collected from 19 food items of read-to-eat foodstuffs which was commonly preferred by Egyptian population and categorized as low price popular foods were investigated for the detection of aerobic colony counts (ACC), coliforms and viable cells of molds and yeasts (M&y) which intensively organized as indicators gives baseline information about the extent of its validity for consumption, and thus, its safety. These samples were collected at a point of sale from multi-premises (small cooking, street-settled stands or mobile cars) prepared such popular foods across Tanta City. Regarding the guideline threshold of Egyptian and Public Health Laboratory Service (PHLS) standards, the results declined that 87% of tested samples did not compliant with standard limits and categorized as unsafe foods for consumption. The nuisance problems associated with the preparing of such foods is undertaken in this research concerning the type of food courses tested the level of target organisms and the temperatures of corresponding food at a point of sampling while they are hold for serving. Also, the other of improper, unhygienic and mishandling approaches inside the premises was observed and noted. This research, in addition, providing a broad discussion including the risk factors associated with the production of such popular foods and the essential issues to assurance its safety in attempts to improve their microbial quality by which could become more peaceable foods for human consumption.

12/20 PURIFICATION AND CHARACTERIZATION OF CANDIDA ALBICANS CHITINASE

Nagwa Tharwat, Mohsen Abou El-Ela and Tahany Abdel-Rahaman

Botany Department, Faculty of Science, Cairo University

Candida albicans secretes an extracellular chitinase when cultivated in medium containing chitin as a sole carbon source. The chitinase was purified to homogeneity by ammonium sulphate, DEAE-Sephadex and Sephadex G-200.   The   enzyme has molecular weight of 60 kDa. The Km and Vmax were 1.4 mg ml-1 and 73.4 μM μg-1 h-1 respectively. The optimum temperature was 40ºC and the optimum pH was 5.5. The enzyme was stable over a wide range of pH (4.5-8) and temperatures (20-50 °C). In assaying the effect of some metal ions, it was found that Mg2+ and Ca2+ stimulated chitinase activity whereas K+ insignificantly activated the enzyme. Cd2+, Ag+ and Hg2+ completely inhibited the enzyme activity. Other tested metal ions inhibited the enzyme activity at different rates.


13/20 ESCHERICHIA COLI O157 IN ACUTE CHILDHOOD DIARRHEA

Safaa M. Abdel-Rahman, Maha M. Fathy, Hanaa M. El-Awady* and

Mervat A. Mohamed

Departments of Microbiology & Immunology and Pediatrics*, Faculty of Medicine,

Ain Shams University

Background: Enterohaemorrhagic Escherichia coli (EHEC) are one of diarrheogenic E.coli that produce shiga toxins causing diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. E. coli O157 is the most common serogroup and the one that can be identified in the clinical specimens of the EHEC species. Aim of Work: The study aimed at evaluating E. coli O157 as a causative agent of acute diarrhea among some Egyptian children, which might help development of optimal management protocols. Methods: We obtained fresh fecal specimens from 60 children presented with acute diarrhea and from 30 healthy controls. Fecal specimens from all subjects were cultured on sorbitol MacConkey’s (sMac) and Cefixime-Tellurite sMac (C-T sMac) agar media. Fecal specimens from patients were cultured also onto selective media for isolation of Salmonella spp., Shigella spp., Yersinia entercolitica and Campylopacter coli/jujeni. Suspected isolates were subjected to identification with standard techniques. Non-sorbitol fermenting and sorbitol fermenting E.coli isolates were serotyped by E.coli O157 latex agglutination test. Results:E. coli O157 was detected in four fecal specimens of children with acute diarrhea (6.7%). The performance of sMac agar media, its sensitivity, specificity, positive predictive value, and negative predictive value were 75%, 96.8%, 75%, and 96.8%, respectively. Yersinia entercolitica was detected in one case (1.7%) of acute diarrhea. No E.coli O157 isolates were detected in the control group. Conclusion: Surveillance for E. coli O157 infection in acute childhood diarrhea is recommended particularly in those with bloody diarrhea. Culture on Cefixime-Tellurite sMac agar media followed by latex agglutination test seems to be a simple and reliable method for E. coli O157 detection.

14/20 MICROBIAL COMMUNITY STRUCTURE IN A WASTEWATER TREATMENT PLANT IN BENI-SUEF CITY

Wael N. Hozzein,Mohammed B. Ahmed* and Marzouka S. Abdel Tawab**

Botany Department and *Biochemistry Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt

**Beni-Suef Wastewater Treatment Plant, Beni-Suef, Egypt

The current study aimed to evaluate the efficiency of the treatment process and to investigate the quantities and diversity of microorganisms in Beni-Suef wastewater treatment plant. For this purpose, samples were collected from 3 different sites representing the main sites in the plant, namely; the influent, secondary sedimentation tanks and the effluent, and the physico-chemical parameters (TS, TSS, TDS, COD, BOD5, DO, O&G, NH3, NO3, NO2, P, Cu, Cr and Pb) in the three sites were examined. All the recorded values indicated good biological treatment in the plant.The viable microbial count in the three sites on three different media; nutrient agar, DOX agar and yeast extract-malt extract agar; was determined. The greatest total number of microorganisms was 3.85x106 CFU/ml in the influent site on nutrient agar, while the smallest total number of microorganisms was 1.34x103 CFU/ml in the effluent site on yeast extract-malt extract agar. It was clear from the enumeration results that nutrient agar was the best medium in revealing both the number and diversity of microorganisms in wastewater. The results showed also that the most active role in the biological treatment was played by representatives of bacteria and actinomycetes, followed by fungi. Among the colonies appeared on the plates, 30 bacterial isolates, 17 actinomycete isolates and 6 fungal isolates were isolated and identified by their cultural, morphological and biochemical characteristics.The bacterial population was dominated by Pseudomonas, Escherichia, Bacillus, Flavobacterium, Staphylococcus, Streptococcus and Xanthomonas. On the other hand, Streptomyces, Nocardia, Nocardiodis, Gordonia and Rhodococcus were the dominant actinomycete genera, while only two fungal genera, Penicillium and Aspergillus, were found.

15/20 BIOACCUMULATION OF HEAVY METALS AND REDUCING OF THEIR TOXICITY ON PLANT GROWTH

Tarek M. Abd El-Ghany

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

Cairo, Egypt.

Response of soil born fungus i.e Aspergillus parasiticus towards heavy metals viz : copper, nickel and cobalt was studied at four different concentrations 10,100,1000 and 1500 ppm. The addition of this metals to the growth medium of A. parasiticus resulted in reduction of relative growth rate and increased the lag phase. Aspergillus parasiticus failed to grow at highest concentrations (1500 ppm) of both copper and nickel while did not for cobalt and also showed maximum accumulation percentage in case of all of these metals at 100 ppm. Bioaccumulation efficiency of Aspergillus parasiticus decreased at high concentrations, 1000 ppm of these heavy metals .The unusual amino acids α-aminobutyric acid, β-aminobutyric acid and β-alanine in addition to histidine and citruline were detected only in the fungus mycelium cultivated in the medium free from heavy metals. On the other hand the amino acid proline was detected in the fungus cultivated in the medium supplemented with heavy metals. The addition of A. parasiticus spores inoculum to agriculture soil supplemented with different concentrations of these metals reduces the sensitivity of cultivated plant (Ficus ntida) to toxic effect of these metals ,and also enhanced plant growth compared with control (plant cultivated in soil supplemented with metals and without fungal inoculum).

16/20 COMPARATIVE SENSITIVITY TESTING OF ASPERGILLUS FLAVUS STRAINS TO MOULD INHIBITORS

Wa`al Tawakkol

Department of Microbiology, Faculty of Pharmacy, Cairo University.

In the present work, ten isolates of Aspergillus flavus were tested for their sensitivity to Formalin, Glutraldehyde, Benzoic, Propionic, Formic and Sorbic acids by determination of minimum inhibition concentration (MIC), colony fungal count and disc diffusion techniques. The average MIC values for all treated isolates were 0.71 for Formalin, and 0.62 for Glutraldehyde treatment. The statistical analysis indicated that the MIC values obtained by Glutradehyde were more uniform than that obtained from Formalin. The average of MIC values obtained from acids application showed that Formic, Propionic and Sorbic acid were in the same order of magnitude (0.68-0.69); while MIC values obtained from Benzoic acid application was the lowest (0.62). The results of colony count revealed that the number of colony after Glutraldehyde treatment was much reduced from the initial count than in case of Formalin treatment. The study indicates the importance of using more than one test and at least 10 isolates of the same fungus in order to get reliable evaluation of any mould inhibitor.

17/20 PULSED-FIELD GEL ELECTROPHORESIS GENOTYPING OF PSEUDOMONAS AERUGINOSA ISOLATED FROM HOSPITALIZED PATIENTS IN ISMAILIA, EGYPT COMPARED TO SUSCEPTIBILITY AND BIOCHEMICAL PROFILES

Sahar M. El-Alfay

Department of Botany, Faculty of Science, Suez Canal University, Egypt

Although DNA-based techniques are the “gold standard” for the epidemiological study of Pseudomonas aeruginosa, antibiotic profiles and biochemical results are used because they are easy to perform and to interpret and relatively inexpensive. Antibiotypes (susceptibility profiles) and biotypes (biochemical profiles) were compared to genotypes established by DNA restriction enzyme analysis by using Pulsed-Field Gel Electrophoresis (PFGE). Twenty clinical isolates of P. aeruginosa recovered from patients admitted to The General Hospital in Ismailia gvernorrate, Egypt, were analyzed in this study. Also P. aeruginosa ATCC (9027) strain was analyzed as a control. The susceptibility tests were performedto represent the different classes of antimicrobial agents commonly used and avilable for the treatment of P. aeruginosa we used aztreonam (ATM 30 μgm), ceftazidime (CAZ 30 μgm), ciprofloxacin (CIP 5 μgm) and imipenem (IPM 10 μgm) Isolates were classified as susceptible or resistant according to the the size of the clear zone. Biotyping was performed by observation of hemolysis after growth on Oxoid blood agar medium for 24 h under transmitted light. Hemolytic isolates were divided into two groups according to observation of α or ß hemolysis. Production of pigment was determined in Mueller-Hinton medium incubated for 24 h at 35ºC, and pigmented colonies were classified according to their color. PFGE. SpeI digestion of the 19 P. aeruginosa isolates from different patients and P. aeruginosa ATCC 9027 control strain were typed by PFGE (one isolate was failed during the analysis), 10 unique patterns (designated pattern types A,A1,B,B1, C, C1,D,D1,E and F) When the genetic relatedness (defined as ≥87% band identity) all strains were grouped in the following ten clusters: two strains in pattern A; two in pattern A1; two in pattern B, two in pattern B1, two in pattern C, patterns C1,D,D1,E each contained single isolate and the five remaining strains were closely related (genomic pattern F).Taking the pulsed-field gel electrophoresis profiles as standard, the same genotype B1 included 2 distinct antibiotype "b" and "d" was observed.Genotypes A,A1,B and C each contained 2 isolates and genotypes D1and E each contained 1 isolates also genotype F which included 5 isolates belonging to the same antibiotype "d". while genotypes C1and D each contained single isolate have their owne uniqueantibiotype "c" and"a" respectively. The genotype pattern of the P. aeruginosa ATCC 9027 control strain and isolate no. 11 was closely related and had two deferent antibiotypes "d" and" respectively. Epidemiological analysis included a variable number of patients and they were considered Epidemiologically related.

18/20 EFFECT OF GROWTH CONDITIONS ON THE PRODUCTION OF ANTIMICROBIAL COMPOUND PRODUCED BY LACTOBACILLUS PLANTARUM AA135, A STRAIN ISOLATED FROM EGYPTIAN DAIRY PRODUCTS

Aly E. Abo-Amer and Bahig A. El-Deep

Division of Microbiology, Department of Botany, Faculty of Science, Sohag University, Sohag, Egypt

Growth of Lactobacillus plantarum AA135 in MRS broth at 30oC yielded maximal plantaricin AA13 activity of 10000 AU/ml at pH 3.8 after 16 hours. Maximal activity (10000 AU/ml) of plantaricin AA13 was recorded in MRS broth at initial pH of 5 or 5.5. Optimal production (10000 AU/ml) was recorded in the presence of tryptone (20 g/l), a combination of tryptone plus meat extract (1:0.6) or tryptone with yeast extract (1:0.6) as sole nitrogen source. Growth of Lactobacillus plantarum AA135 in the presence of 20 g/l glucose yielded bacteriocin activity of 10000 AU/ml. Moreover, when 40 g/l glucose was added, the plantaricin AA13 levels doubled to 20000 AU/ml. Concentrations of 5, 10 and 20 g/l K2HPO4 lowered bacteriocin activity by 60%. Supplementing MRS with 1 g/l or more glycerol repressed bacteriocin production. Maximal activity of 20000 AU/ml was recorded in MRS supplemented with Vit. DL-6,8-thioctic acid.

19/20 EFFECT OF MICROBIAL INOCULANTS, MINERAL AND/OR ORGANIC FERTILIZERS ON PRODUCTIVITY, NUTRIENT STATUS AND SOIL MYCOBIOTA OF FLAME SEEDLESS GRAPEVINES

Nemmat A. Hussein and Yasser A.M.M. Omran*

Botany Department, Faculty of Science, Assiut University, Egypt;

*Viticulture Department, Horticulture Research Institute, Giza, Egypt;

A study was conducted to investigate the effects various amounts of three types of fertilizers {mineral (M), farmyard manure (FYM) and microbial inoculants (EM1)} on vegetative growth, productivity and soil mycobiota of cv. Flame Seedless grapevines in the 2004 and 2005 seasons. The results indicated that all treatments except FYM and / or EM1 increased vegetative growth and leaf chlorophyll content compared to the control (mineral fertilizer at the recommended rate). EM1 addition to FYM fertilizer enhanced elemental levels in leaves and addition to mineral fertilizer significantly increased elemental content. Furthermore, its addition to (100% M + 100% FYM) caused highly significant increases in the nitrogen, magnesium, iron, zinc and manganese contents of leaves. 50% M + 50% FYM + EM1 significantly increased yield weight while, farmyard manure and EM1, alone or together, significantly decreased yield weight by about 12%, 28% or 9.5%, respectively. The remaining treatments improved yield components and all treatments also led to the enhancement of fruit quality. The mycological diversity in the soil was investigated after addition of treatments 4 weeks after harvest. Fungi were enumerated and identified on Czapek Dox agar at 28ºC. Total fungal counts in all samples tested ranged from 24,400 – 138,400 colonies / g soil; the highest counts were found in treatments with 100% M + 100% FYM, mineral fertilizer, 50% M. + 50% FYM and EM1, while the lowest counts were found in treatments with 25% M. + 75% FYM + EM1, FYM, 25% M. + 75% FYM, 75% M. + 25% FYM, and 100% M. + 100% FYM + EM1. Aspergillus (A. niger, A. japonicus, A. terreus and A. flavus), Penicillium (P. oxalicum, P. pinophilum and P. purpurogenum) and Fusarium (F. scirpi and F. sambucinum) were the most prevalent fungi.

20/20 PLASMID INCIDENCE FLUCTUATIONS IN BACTERIAL POPULATION FROM AGRICULTURAL SOIL AT SOHAG PROVINCE, EGYPT

Bahig A. El-Deep, Aly E. Abo-Amer*, Khaled A. Ali and Amel K. Ahmed

Division of Microbiology, Department of Botany, Faculty of Science, Sohag University, Egypt

Forty soil samples of agriculture soil were collected from two different sites in Sohag province, Egypt, during hot and cold seasons. Twenty samples were from soil irrigated with canal water (site A) and twenty samples were from soil irrigated with wastewater (site B). This study aimed to compare the incidence of plasmids in bacteria isolated from soil and to investigate the occurrence of metal and antibiotic resistance bacteria, and consequently to select the potential application of these bacteria in bioremediation. The total bacterial count (CFU/gm) in site (B) was higher than that in site (A). Moreover, the CFU values in summer were higher than those values in winter at both sites. A total of 771 bacterial isolates were characterized as Bacillus, Micrococcus, Staphylococcus, Pseudomonas, Eschershia, Shigella, Xanthomonas, Acetobacter, Citrobacter, Enterobacter, Moraxella and Methylococcus. Minimum inhibitory concentrations (MICs) of Pb+2, Cu+2, Zn+2, Hg+2, Co+2, Cd+2, Cr+3 Te+2, As+2 and Ni+2 for plasmid-possessed bacteria were determined and the highest MICs were 1200 mg/ml for lead, 800 mg/ml for both Cobalt and Arsenate, 1200 mg/ml for Nickel, 1000 mg/ml for Copper and less than 600 mg/ml for other metals. Bacterial isolates from both sites A and B showed multiple heavy metal resistance. A total of 337 bacterial isolates contained plasmids and the incidence of plasmids was approximately 25-50% higher in bacteria isolated from site (B) than that from site (A). These isolates were resistance to different antibiotics. Approximately, 61% of the bacterial isolates were able to assimilate insecticide, carbaryl, as a sole source of carbon and energy. However, the Citrobacter AA101 showed the best growth on carbaryl.

21/20 SHRIMP SHELL AS AN INEXPENSIVE SUBSTRATE FOR PROTEASE PRODUCTION BY PSEUDOMONAS ALCALIGENES IN SUBMERGED FERMENTATION

Sherif M. Husseiny, Fatma A. Halamsh and Rasha M. Said

Botany Department, Women's College for Arts, Science, & Education,

Ain Shams University

Protease production under submerged fermentation (SmF) was investigated using Pseudomonas alcaligenes. This strain was isolated from fermented shrimp shell wastes for 15 days in open air. Protease was produced by growing the isolated strain on a medium containing shrimp shell powder as a carbon and nitrogen sources. Optimum enzyme production was achieved at 40ºC for 30 h and initial pH 7 with 0.6 g dry shrimp shell waste. Inoculum size, yeast extract concentration, nitrogen sources played a vital role in protease production

22/20 MOLECULAR AND BIOCHEMICAL STUDIES ON PATHOGENIC STREPTOCOCCI FOR ISOLATION AND CLONING OF STREPTOKINASE GENE

Mahmoud Abdel-Mongy, Nasser H. Abbas, Haddad A. El Rabey,

Hassen El Berry*, Bakry Haroun* and Khalil A. El Halafawy

Genetic Engineering and Biotechnology Institute, Minoufiya University, Sadat City, Egypt.

*Faculty of Science, Al Azhar University, Cairo Egypt.

Molecular genetic techniques have been applied on several streptococcus isolates from different habitats in order to investigate the presence of streptokinase enzyme that has medical importance in dissolving blood clots. Several isolates prove their production of active streptokinase. The Plasminogen zymography show the expected molecular weight of 45 kDa of the streptokinase enzyme. The zymgraphy demonstrated also the correlation between the enzyme and the plasmogene substrate of its host. The total cell protein profile of the isolated strains was determined. specific primers was designed and used to isolate complete gene sequence from the defined isolate. The isolated gene was recombined into cloning vector and transformed into E. coli.

23/20 DETECTION OF EPSTEIN BARR VIRUS IN CASES OF HEMODIALYSIS AND RENAL TRANSPLANT RECIPIENTS AND ITS POSSIBLE IMPACT AFTER ONE YEAR FOLLOW UP

Hoda Kholeif* and Sahar Shaweky

*Clinical Pathology Department, Faculty of Medicine for Girls, Al-Azhar University

Nephrology Department, Faculty of Medicine, Ain Shams University

Epstein Barr virus (EBV) causes a number of conditions including a variety of human malignancies. Post transplant lymphoproliferative disorders are nearly always associated with EBV and the use of immunosuppressive drugs. The aim of this study is to determine the incidence of EBV-DNA in a group of hemodialysis patients and renal allograft recipients and its possible impact on patients in one year follow up period. This study was conducted on 60 subjects who were divided into three groups, 25 hemodialysis patients with different causes of renal failure, 20 renal allograft recipients attending outpatient clinic for follow up and 15 blood donors attending the blood bank of Ain Shams university hospitals. Three milliliters of EDTA anti-coagulated whole blood were obtained from each person included in this study. Detection of human EBV-DNA was carried on peripheral blood mononuclear cells (PBMC) by polymerase chain reaction ( PCR ): EBV-DNA was detected in 14 out of 25 hemodialysis patients (56%), 9 out of 20 renal allograft recipients (45%) and in none of the blood donor group (0%). There was no relation found between age, sex, and being positive for EBV-DNA. After clinical follow up for one year, one of the EBV-DNA positive renal transplant recipient developed complication in the form of Hodgkin’s lymphoma. In conclusion EBV is one of the most important risk factors for the onset of post transplant lymphoproliferative disorders (PTLD), also subclinical asymptomatic EBV viremia is a risk factor for graft injury and loss in renal transplant recipients.

24/20 RESPONSE OF DIFFERENT ANTIBIOTIC RESISTANT GROUP OF STREPTOCOCCUS PYOGENES TO ENVIRONMENTAL STRESSES

Nasser H. Abbas, Mahmoud Abdel-Mongy, Mohamed El-Shahat, Hassan El Berry* Bakry Haroun* and Khalil A. El Halafawy

Genetic Engineering and Biotechnology Institute, Minoufiya University,

Sadat City, Egypt.

*Faculty of Science, Al Azhar University, Cairo Egypt.

Streptococcus species is considered as an important pathogen for human and animals. The antibiotic resistance mechanism in this species is continuously increased. On the other side, the environmental stresses play an effective role in the severity of many streptococcal causative disease. In this study we assayed survey on the causative agents of pharyngitis and tonsillitis patients. The predominant causative strain was Streptococcus pyogenes with 93% isolating ratio frequency. The other pathogenic species were S. agalactia 32%, S. pneumonia 17%. According to the antibiotic resistant test the S. pyogenes isolates were classified into 6 different groups. A selected strain from each antibiotic resistant group was tested for tolerance of a restrictive environmental factors.

25/20 ANTIFUNGAL ACTIVITY OF SOME MEDICINAL PLANT EXTRACTS ON DIFFERENT PHYTOPATHOGENIC FUSARIUM SPP. WITH SPECIAL REFERENCE TO CONTROLLING WILT DISEASE OF TOMATO PLANTS

Omyma A. Awadalla

Botany Department, Faculty of Sciences, Tanta University

The antifungal activity of air dried leaves of five Egyptian medicinal plants was investigated; these plants were eucalyptus (Eucalyptus globulus), garlic (Allium sativum), lemon grass (Cymbopogon citratus), marjoram (Marjorana hortensis) and peppermint (Mentha piperita). Aqueous extracts of these plants were tested against six phytopathogenic Fusarium spp. (F.oxysporum f.sp.lycopersici, F.solani. F.oxysporum f.sp. vasinfectum, F.culmorum, F.moniliforme and F.graminearum. All aqueous extracts of medicinal plants exhibited a broad fungitoxic spectrum by inhibiting the mycelial growth of all the tested Fusarium spp. in solid and liquid media. The degree of inhibition was directly proportional with the concentrations of plant extracts. The linear mycelial growth of F.oxysporum f.sp.lycopersici, F.moniliforme, F.solani and F.culmorum was completely inhibited at 10% concentration of all the used extracts. Using plant extracts in liquid media were more effective in reducing the mycelial dry weight of all tested Fusarium spp. comparing to the solid media, where all plant extracts at 5% and 10 % concentrations completely inhibited the mycelial growth of all tested Fusarium spp. It was observed that F.oxysporum f.sp.lycopersici was the most affected by the used plant extracts compared to the other used Fusarium spp.. Accordingly it was chosen for in vivo study to control Fusarium wilt disease of tomato using the tested plant extracts. In this work, the results revealed that extracts of eucalyptus, garlic and lemon grass were the most effective in controlling the disease at all used concentrations, where the wilt disease symptoms were completely disappeared. Leaf extracts of peppermint and marjoram were also effective in controlling wilt disease of tomato; they could completely control the disease at the two highest concentrations.

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