Vol. 29, June, 2008.

cialis diabetes effets secondaires 1/29 ANTIMICROBIAL ACTIVITY OF BIOCHEMICAL cialis pas cher espagne CONTENTS OF SPIRULINA PLATENSIS

Awatef E. Ghoniemy, Howayda H. Abd El-Hady and Said M. Daboor

Hydrobiology Laboratory, National Institute of Oceanography and Fisheries "NIOF", Cairo, Egypt.

The biochemical contents of cialis pas cher en pharmacie lyon Spirulina platensis was analyzed. The results indicated that commande cialis par internet S. platensis is a remarkably rich with of proteins, pigments, vitamins and growth factor, it contain high concentrations of protein (60%), moderate contents of carbohydrate and lipid (14 and 3.6 % respectively), β-carotene (5.68 g/kg dry weight "DW") and chlorophyll “a” was 0.601 mg/g DW. Water and ethanol extracts of levitra ou cialis effet S. platensis were tested on some microorganisms, two Gram positive, two Gram negative bacteria, two fungi and one yeast. Whole powder was effective against all the tested microorganisms at concentration 15% (w/v), while water extract was not, on the other hand the ethanolic one was active against all the tested microorganisms. The minimum concentration of the extract varied with the organism where nafil sildenafil plus Staphylococcus aureus NIOF001 was the most susceptible isolate.

durĂ©e traitement levitra  

viagra en ligne au quebec 2/29 INFLUENCE OF SALINITY ON BIOMASS PRODUCTION, PHOTOSYNTHETIC PIGMENTS AND PROLINE CONTENT OF TWO FODDER BEET CULTIVARS

Samya M. Sohsah

Botany Department, Faculty of Science, Al-Azhar University (Girls)

The present work intends to evaluate the influence of salinity treatment (Zero, 4000, 8000 and 12000 ppmNaCl: Ca Cl2; 2: 1 w/w) on growth and metabolic compounds of two fodder beet ( medicament similaire au viagra Beta vulgaris L.) varieties (Magnum and Monored) during various growth stages. Salinity at any applied level induced significant decrease in all tested aspects of plant growth; including plant height, fresh weight of shoot and root, dry weight of shoot and root, number of leaves / plant, leaf area and root width. The magnititude of such decrement was much higher in Monored variety than in Magnum variety. Meanwhile, a pronounced increase in root length of the two fodder beet varieties under different salinity levels was detected throughout the growth period. All growth aspects also increased with the advancement of plant age, while salinity induced a depressive effect on chlorophyll "a" and "b", but showed an enhansive effect on carotinoid and proline content in leaves of the two tested varieties of fodder beet.

finasteride contre la chute des cheveux 3/29 Transposition of Both Trigonella cylindracea DesV. and T. polyceratia (l.) trautv. to genus Medicago as inferred by RAPD and RFLP analyses

Ream I. Marzouk and Ranya El-Bakatoushi*

Department of Botany, Faculty of Science, Alexandria University

*Department of Biological and Geological Sciences, Faculty of Education,

Alexandria University

The taxonomic relocation of both Trigonella cylindracea Desv. and T. polyceratia (L.) Trautv. either to Trigonella or Medicago is assessed on the basis of two molecular markers, RAPD and RFLP. Through RAPD analysis, 109 bands are generated by ten random primers with an average 11 products/primer.   The maximum number of RAPDs (76 bands) associated with highest percentage of polymorphism (43.3%) are recorded with T. polyceratia. While, the minimum records combined with T. foenum-graecum, 63 bands and 32.3%. RFLP analysis of 18S rDNA is performed by using five restriction endonucleases (EcoRI, HindIII, BamHI, HinfI and TagI).   The restriction endonuclease HinfI and TagI produce two and three fragments respectively and no variation is observed in these sites among the studied species. However, another three restriction endonuclease (EcoRI, HindIII& BamHI) are undigested. The present study suggested the reallocate of both T. cylindraceae and T. polyceratia to Medicago and certified the value of RAPD analysis for the taxonomic intention.

4/29 EVALUATION OF THE NUTRITIONAL COMPOSITION OF SEVEN SEAWEEDS FROM EGYPT

Islam M. El-Manawy

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

The nutritional composition of dried samples of two red seaweeds (Gracilaria disticha and Hypnea valentiae, one brown (Dictyota dichotoma) and four green (Caulerpa lentillifera, Enteromorpha flexuosa, Ulva fasciata and Ulva lactuca) from the Egyptian coasts were determined. Total dietary fiber was the most abundant component in these algae (33.31 ± 0.18 to 49.06 ± 0.82 g.100g-1), followed by ash content (22.49 ± 0.08 to 27.16 ± 0.28 g.100g-1) and crude protein (12.3 ± 0.73 to 32.71 ± 0.68 g.100g-1). The ratio of soluble fiber fraction to the total fiber was between 49-64 %. Available carbohydrates were 7.07 ± 0.62 to 14.77 ± 1.14 g.100g-1 and total lipids were (2.13 ± 0.05 to 6.23 ± 0.41 g.100g-1). Gross energy was estimated between 138 ± 16 and 186 ± 15 Kcal g.100g-1. Moisture contents were from 12.00 ± 0.74 to 16.33 ± 0.52 g.100g-1. Results show that the red species characterized by higher proteins with lower lipids, carbohydrates, and fibers; whereas the contrary was found for green and brown species.

 

5/29 A PROMISING 2–DEOXYSTREPTAMINE AMINOCYCLITOL AMINOGLYCOSIDE ANTIBIOTIC FROM A STREPTOMYCES GLOBOSUS ISOLATE WITH ANTIBACTERIAL ANTIFUNGAL ACTIVITIES

Walid F. El-Khatib, Mohammad M. Aboulwafa, Mahmoud A. Yassien

and Nadia A. Hassouna

Department of Microbiology and Immunology, Faculty of Pharmacy,

Ain Shams University

A screening program applied on 730 Streptomyces soil isolates for their antimicrobial activities resulted in the recovery of the isolate S460 that exhibited the highest antimicrobial activities against the tested Gram-positive and Gram-negative bacteria as well as fungi relative to the other Streptomyces isolates. The growth supernatant of this isolate yielded one and two spots with antimicrobial activities on paper chromatography using 60% v/v acetic acid and distilled water as mobile phases, respectively. Also, the growth supernatant of this isolate showed the lowest percentage of mammalian cell lines destruction compared to other tested isolates and some known antibiotics by cytotoxicity assays. The isolate S460 was identified as Streptomyces globosus and selected for antibiotic production. The antibiotic in the growth supernatant of this isolate was extracted, purified by TLC chromatography and then crystallized. When the crystallized antibiotic was re-chromatographed on cellulose paper and the developed chromatogram was bioautographed, the results revealed the existence of a single antibiotic which is active against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa and Candida albicans ATCC 60193. The thermal stability, physical and chemical properties as well as spectroscopic analyses revealed that the antibiotic produced by Streptomyces globosus S460 can be considered as a new aminoglycoside antibiotic with 2-deoxystreptamineaminocyclitol basic structure.

 

6/29 OPTIMIZATION OF MEDIUM COMPOSITION FOR THERMOSTABLE XYLANASE PRODUCTION BY ASPERGILLUS NIGER INSOLID STATE FERMENTATION

Rania M.A. Abedin

Botany Department (Microbiology), Faculty of Science, Alexandria University, Alexandria, Egypt.

The enzyme named xylanolytic deconstructs plant structural material by breaking down hemicellulose, a major component of the plant cell wall. Many micro-organisms produced xylanase. Aspergillus niger was examined for the production of xylanase using solid state fermentation technique on corn cobs and many other solid substrates. Optimization of the SSC medium for the production of thermostable xylanase by Aspergillus niger in SSC medium at pH 6.5 with corn cobs as substrate after 5 days of incubation yielded 6.36 U/ml. Also, the effect of some reaction conditions on the activity of crude xylanase were studied resulted in 1.37-fold increased level of thermostable xylanase (8.71 U/ml) production compared to initial level (6.36 U/ml).The optimum substrate concentration of the crude xylanase was found to be (0.2 g/100ml). The optimum temperature of the activity was found to be 55oC at an optimum pH 5.6. The enzyme remained stable up to 60oC, yet lost about 48% of its original activity. The strain under test have a good potential for use as a source of important industrial enzyme of high thermal stability.

7/29 PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR LIPASE FROM FUSARIUM MONILIFORME

Amal A. Khalil and Medhat A. Ali

Department of Biological Science, Faculty of Education,

Ain Shams University, Heliopolis, Roxy, Cairo, Egypt.

An extracellular lipase from Fusarium moniliforme was obtained when grown on a medium contained triolein as a sole carbon source. The enzyme was purified 5.65 fold with total recovery of 14.52% through purification procedure of ammonium sulfate precipitation, gel filtration through Sephadex G-100 and ion exchange chromatography on DEAE- Cellulose. Maximum enzyme activity was recorded at pH 7 and 35ºC. The enzyme has a good stability at pH values ranged between 6 and 7. A high degrees of stability was recorded at temperatures less than 40ºC. Km value of the purified enzyme was calculated to be 0.22 mM. Enzyme activity was enhanced in presence of Na+, Ca+2 and Mg+2 at the time that Hg+2, Fe+2, Fe+3 and Zn+2 caused strongly inactivation, on the other hand 1 or 10 mM of EDTA and 1 mM of 2,4, dinitrophenol did not affect the enzyme activity. The apparently purified enzyme showed the highest activity on triolein. Sharing of carbohydrate moiety of the enzyme protein has been confirmed that it is a conjugated protein (glycoprotein). The enzyme was stable for 25 days when kept at 4ºC and -15ºC. T1/2 of enzyme activity was found to be 49 days at the two stated temperatures.

 

 

8/29 PRODUCTION OF EXTRACELLULAR ALKALINE PROTEASE BY EXTREMOPHILE BACTERIAL STRAINS ISOLATED FROM EGYPTIAN COASTAL ALKALINE SALTY SANDY SOILS

Ola A. Abd-Elrahman, Abeer K. Abdulaal and Tarek Kahil*

Department of Microbiology & Immunology, Faculty of Pharmacy, Azhar University and *Microbial Chemistry Department, Genetic Engineering & Biotechnology Research Division, National Research Center, Cairo, Egypt

The present study is focused on isolation of extracellular alkaline protease (AP) producing extremophilic bacteria, halophiles or alkalophilesorhaloalkalophiles,screening of these isolates for the highest AP-producers, and studying some production affecting factors. Thus, five samples of local coastal alkaline sandy soils from Suez Canal Coast (szc) - Red Sea & Western North Coast (wnc) - Mediterranean Sea were incubated in 4 isolation media of the same constituents but differing at pH and salinity; i.e., with 2 alternative pH values (7 or 10) & 2 alternative concentrations of NaCl (0 or 3%). Twelve protease-producing bacterial strains were isolated from colonies developed on these media. They were categorized into four groups: (i) Four Neutra-nonhalophilic (N.nh) strains; all isolated from Western North Coast and identified as Streptomyces albidoflavus (strains wnc1/N.nh & wnc3/N.nh), Lactococcus lactis supsp. lactis (formerly Streptococcus lactis) wnc3/N.nh and Arthrobacter agilis (formerly Micrococcus agilis) wnc2/N.nh; (ii) Three Alkalo-nonhalophiles (A.nh) isolated from Suez Canal Coast (Bacillus acidocaldarius szc1/A.nh) and Western North Coast (B. azotoformans wnc1/A.nh & Micrococcus roseus wnc2/A.nh); (iii) Three Neutra-Halophiles (N.H) isolated from Suez Canal Coast (B. azotoformans szc1/N.H & Methylococcus capsulatus szc2/N.H) and Northern Western Coast (M. capsulatus wnc1/N.H) and (iv) Two Alkalo-Halophiles (A.H) isolated only from Suez Canal Coast (Pseudomonas putida szc1/A.H & Methylococcus capsulatus szc2/A.H). The highest AP activities exhibited by different strains were, in descending order, as follows: Actinomycetes [Streptomyces albidoflavus wnc3/N.nh (96U on 7th day) & S. albidoflavus wnc1/N.nh], Methylococcus capsulatus szc2/N.H, B. azotoformans szc1/N.H, Lactococcus lactis, Arthrobacter agilis, Methylococcus capsulatus wnc1/N.H, Pseudomonas putida, Methylococcus capsulatus szc2/A.H, B. acidocaldarius, Micrococcus roseus and B. azotoformans wnc1/A.nh. There was no relation between maximum productions of alkaline protease and either alkalinity or salinity of isolation & production media. Genera, species, & strains of isolated organisms have affected greatly AP production & time at which the yield was maximum. On the other hand, Gram reaction had no impact on AP production. Relation between culture final-pH & AP production was positive in 7 strains and negative in 4 strains. Whereas, there was no relation in case of Pseudomonas putida. Thus, the effect of genera, species, and strains on this relation was obvious. On the contrary, productions of AP by all strains investigated were positively correlated with growth (as biomass). No relation was found between the maximum AP attained and experimental-culture age.

 

9/29 Purification and Biochemical Characterization of Alkaline Protease from Biopesticide-Producing Strain of Bacillus sphaericus

Abd El-Moneim M.R. Afify, Mohamed S. Foda*, Murad A.M. Aboul-Soud, Mahmoud A.M. Mohamed, Tarek Kahil* and Amira R. Asar*

Biochemistry Department, Faculty of Agriculture, Cairo University, Giza, Egypt

*Microbial Chemistry Department, National Research Center, Giza, Egypt

The present work was advocated to purify and elucidate the biochemical properties of the purified alkaline protease (AP) enzyme from biopesticides-producing strain of Bacillus sphaericus no (5) isolated from Egyptian soil. This study is needed to evaluate its potential as AP enzyme producer as an added value to the capacity of producing useful biopesticide in the same fermentation process. The crude alkaline protease of B. sphaericus was purified 50 folds using purification scheme based upon ammonium sulfate fractionation followed by fractional precipitation with acetone and gel filtration chromatography on Sephadex G-100 column. The highest enzyme activities were obtained in ammonium sulfate precipitated fraction between 30-60% saturation levels. This step was followed by acetone precipitation, with highest enzyme activity obtained in acetone-precipitated fractionation in the range between 50-75% v/v. The active fraction was applied to Sephadex G-100 column with glycine-sodium hydroxide buffer (0.1M, pH 10) was used for enzyme elution from the column. Forty-eight fractions (5ml each) were collected. Most of AP activities were eluted within seven fractions numbering between 13 & 19 with total eluate volume of 35 ml containing the purified AP activity. The AP activity was thus purified 49.5 folds as compared to the original crude enzyme with yield recovery 27%. Biochemical characterization of purified AP revealed that the maximum activity of the enzyme was at pH 10 with slight decrease in activity up to pH 12 of the same buffer. On the other hand, the AP was thermolabile to temperatures higher than 40ºC, in the absence of casein as a substrate. Under these conditions, the optimum incubation temperature for maximal AP activity ranged between 55-60ºC, indicating the substrate (casein) protection effect on enzyme activity. The rate of the enzyme-catalyzed proteolysis was linear with lower enzyme concentrations. The purified AP was slightly activated in the presence of low concentrations (1mM) of cations namely calcium, manganese, cobalt, nickel and ammonium, in the form of chloride or sulfate salts. On the other hand, clear inhibitions of the AP activity were recorded in the presence of ferrous or mercury salts and the inhibitions were progressively increased by increasing their concentrations and to a lesser extent with zinc in the same forms of salts & concentrations. Concerning the effects of different anions (in the form of sodium salts), formate, carbonate and monobasic phosphate anions, at the same level (1mM), exerted notable stimulation of the purified AP activity at low concentrations, with little or no effect for chloride, acetate and citrate anions. Maximum enzyme activity was recorded upon using casein as a substrate with Vmax value about 230μg tyrosine/ml/min and Michaelis constant Km value of 0.05%, w/v, of casein substrate. Different levels of activities were obtained for other substrates including skim milk, gelatin, bovine serum albumin, and azocasein. The results were discussed in the light of the biochemical properties of other bacterial alkaline proteases reported in the literature.

 

10/29 EVALUATION OF ANTIBACTERIAL, ANTIFUNGAL AND ANTIVIRAL ACTIVITIES OF TEN MARINE MACROALGAE FROM RED SEA, EGYPT

Neveen Abdel-Raouf, Ibraheem B.M. Ibraheem, Mohamed S. Abdel-Hameed and Khaled N.M.E. El-Yamany

Botany Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt.

Diethyl ether, acetone and ethanol extracts of ten marine macroalgae; two belonging to Chlorophyceae (Ulva lactuca and Caulerpa racemosa), two belonging to Rhodophyceae (Acanthophora spicifera and Galaxaura elongata) and six taxa belonging to Phaeophyceae (Liagora farinosa, Cystoseira compressa, C. myrica, Hydroclathrus clathratus, Turbinaria ornata and Padina pavonia) which isolated from the inter tidal zone along Qusier Marsa-Alam seashore (Red Sea), Egypt, were evaluated for their antibacterial, antifungal and antiviral activities against 3 Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus and Sarcina maxima), 3 Gram-negative bacteria (Pseudomonas aeruginosa, E. coli and Klebsiella pneumonia), one unicellular fungus Candida albicans and two filamentous fungi (Aspergillus flavus and Fusarium oxysporum) and against the New Castled sense Virus (NDV)-(Paramyxoviridoe) which is responsible for acute respiratory distress in chicken. Data showed that some extracts recorded strong inhibitory activities rather than reference antibiotics. Others with moderate and/or week inhibitory activities. However, many without any inhibitory effects. The cytotoxicity effect of the tested algal extracts on chicken embryo showed that both diethyl ether and acetone extracts had toxic effects, but the ethanol extract had no toxic effect. So that ethanol extract was considered to be the most suitable for further studies. The antiviral activities of the ethanol algal extract against NDV (New Castle Disease Virus) showed that seven of the ten tested algal extracts have strong activities against NDV.

 

11/29 GROWTH AND DEVELOPING OF DATE PALM PLANTLET FOR PRE- ACCLIMATIZATION STAGE: THE EFFECT OF GROWTH RETARDANTS AND VESSEL TYPES.

Ezz G. Gadalla; Rasmia S. Darwish; Rehab A. Sidky and Zeinab E. Zaid.

Central Laboratory for Date Palm Research and Development, Agricultural Research Center Giza, Egypt.

This study has been carried out at Central Laboratory for Research and Development of Date Palm, Agriculture Research Center (ARC) Giza, during 2007-2008 on Phoenix dactylifera cv. Malakaby (as a dry cultivar) to investigate the effect of some growth retardants (Abscisic acid (ABA), Paclobutrazol (PBZ) and Ancymidol (ANC) ) with different at concentrations each (0. 0, 0. 5, 1. 0, 1. 5 and 2. 0 mg/l) and type of containers culture jar (350 ml) and flask (250 ml) on vegetative and rooting growth of plantlets, in order to improve the trunk thickness of plantlets, accelerate root formation and increase root branching (secondary roots) for successful transplanting. This study showed that, using flask (250 ml) covering by aluminum foil gave the best results in all parameters compared with using jars 1.0 or .05 mg/l ABA recorded higher value in number of roots than other treatments. Ancymidol at .05 mg/l increased significantly the root length, while paclobtrazol reduce the root length. 2.0 mg/l Ancymidol increased significantly the plantlet thickness. ABA at 0.5 mg/l and paclobtrazol at 1.0 mg/l increased significantly the content of phenols and indols. Rooted plants have been successfully acclimatized with a success rate of around 93%.

12/29 DETECTING THE EXPRESSION OF A MICROCYSTIN BIOSYNTHETIC GENE (MCYB) FROM MICROCYSTIS STRAINS AND THE EFFECT OF TEMPERATURE ON THE EXPRESSION USING REAL-TIME PCR

Nermin A. El Semary

Department of Botany and Microbiology, Faculty of Science, University of Helwan, Ain Helwan Campus, Egypt.

Microcystins, toxins produced by some cyanobacterial taxa, including Microcystis, pose a real problem in freshwater reservoirs used for drinking, fish farms and irrigation. The toxic effect can critically influence the public health where there is a direct link between these toxins and tumor induction and/or organs damage. Classical methods are not useful in identifying toxic strains and the use of advanced techniques is required. The study here used an efficient method for detecting the potential of toxin production and its expression from both cultures and environmental samples of Microcystis through the amplification of one of the toxin biosynthetic genes (mcyB) and the reverse transcription of this gene. A protocol for the quantification of the expression through the use of real-time PCR was developed. This molecular approach verified the toxin production by some Microcystis strains and proved to be very efficient in distinguishing toxic from non toxic Microcystis strains. Moreover, the detection of expression of the toxin biosynthetic genes was used in defining the effect of temperature on toxin production. The toxin production increased significantly as the temperature rose from 18 to 25ºC. Therefore, the quantification of toxin biosynthetic gene expression is quite useful for drawing a policy for toxic bloom control.

 

 

13/29 ANTIFUNGAL ACTIVITY OF ETHNO MEDICINAL PLANT EXTRACTS FROM AL-BAHA FORESTS ON SOME MYCOTOXINS PRODUCING FUNGI

Mohamed A. Foaad

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

Continuing field interviews brought the total species used for disease treatment by Bedouin of Al-Baha region in king of Saudi Arabia. Literature searches provided support for the ethnomedical claims for a number of these species, and provided criteria for the most and safer ones which could use in fungal infections. Crude oil exudates from Psiadia punctulata and stem & leaves extracts of Teucrium polium were chosen and   examined for antifungal activity against mycotoxigenic fungi. Aspergillus flavus (Aflatoxins producer), Penicillium expansum (patulin producer) and Fusarium moniliforme (Fumonisin B1 producer) were underwent. Different fractions were prepared from the leaves & stems of Teucrium polium and were tested for MIC-100 for the tested fungi. The concentrations that lead to complete inhibition of fungal growth (MIC-100) were measured by sensitive XTT assay. MIC-100 of crude oil was 10µl/100µl DMSO for both Aspergillus flavus and Penicillium expansum and 15 µl/100µl DMSO for Fusarium moniliforme. MIC-100 of the fractions showed variable values depending on the solvent used in the fractionation. Methanol, chloroform, ethanol and ethylacetate fractions showed higher activity with lower MIC-100 than petroleum ether and water fractions which showed lower activity. The antifungal activity of plant materials fractions and crude oil were fungus dependent. The inhibitory activity of the extracts on each fungus was monitored by measuring the percentage of spore germination. The inhibitory activity of the plant materials were inoculums dependent. As the inoculums concentration increased the MIC-100 of the plant materials increased. No inhibitory activity was documented at the high concentrations of Fusarium moniliforme spores within the concentration range of tested materials. Both crude oil of Psiadia punctulata and different fractions of Teucrium polium at MIC-100 were subjected to toxicity test on seed vitality by lettuce seedling germination assay .Fortunately, all fractions and crude oil at their highest values of MIC-100 were non toxic for seed germination which showed no significant difference between the control and treated experiments.

 

 

14/29 OPTIMUM CONDITIONS FOR URICASE ENZYME PRODUCTION BY GLIOMASTIX GUEG

Atalla, M.M., Farag, M. M*, Eman R. Hamed, Abd-El-Lataif, M.S.* and Nehad E. Ahmed

Department of Microbial Chemistry Products, National Research Center (NRC), Dokki and *Depatment of Biochemistry, Faculty of Agriculture, Cairo University, Giza, Egypt

Nineteen strains of microorganisms were screened for uricase production. Gliomastix gueg was recognized to produce high levels of the enzyme. The optimum fermentation conditions for uricase production by Gliomastix gueg was examined. Results showed that uric acid medium was the most favorable one, the optimum temperature was at 30ºC, incubation period required for maximum production was 8days with aeration level at 150 rpm and at pH 8.0. Sucrose proved to be the best carbon source, uric acid was found to be the best nitrogen source. Both, di potassium hydrogen phosphate and ferrous chloride as well as some vitamins gave the highest amount of uricase by Gliomastix gueg.

 

15/29 ELICITATION OF TRANS-RESVERATROL BIOSYNTHESIS IN EGYPTIAN GRAPE BERRIES BY THE   ELICTOR FROM CULTURE FILTRATE OF ASPERGILLUS CARBONARIUS

Mohamed A. Foaad and Ahmed F. Afifi

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

The mycological analysis of fifteen seedless grape berries collected from Egyptian market and fields showed that Aspergillus carbonarius is the most strain contaminate the grape berries and producing ochratoxin A. The relationship between the contamination of grape berries by this fungus and elicitation of Trans-resveratrol biosynthesis were studied. Three day old culture filtrate of the fungus stimulated the grape berries to synthesis Trans-resveratrol In vitro. The culture filterate was subjected to purification by means of gel filtration chromatography using Sephadex G-50, 75 and 100 in order to detect the presence of elicitor. The fractions of the purified elicitor were investigated to elicitation of Trans-resveratrol biosynthesis in grape berries. The elicitor fractions purified by Sephadex G-100 were the most stimulatory activity for Trans-resveratrol biosynthesis. The production of Trans-resveratrol under the effect of the purified elicitor was monitord by phenylalanine ammonia lyase as a biomarker for Trans-resveratrol biosynthesis Different factors were investigated to study the nature and stability of the purified elicitor. The electrophoretic profile of the elicitor revealed that is a protein of forty bands. The conclusion is that the present study needs further genetic study in the future to detect the molecular regulation of elicitor synthesis by the fungus.

 

 

16/29 IMPROVEMENT OF CRUDE PETROLEUM-OIL BIODEGRADATION BY PSEUDOMONAS AERUGINOSA JW-1 USING RHAMNOLIPID

Haytham M. M. Ibrahim

Department of Microbiology, National Center for Radiation Research and Technology, Atomic Energy Authority, Nasr city, Cairo, Egypt.

The aim of the present study was to find a method to enhance crude petroleum-oil biodegradation by Pseudomonas aeruginosa JW-1. This strain was reported to produces an abundant amount of rhamnolipid, when provided by vegetable oils, but it failed to utilize crude oil directly as a sole carbon source i.e., neither cell growth nor rhamnolipid production was observed. The results showed that the bacterial culture slightly emulsified crude oil, produced little amount of rhamnolipid (0.33 g/l) and achieved 37.5 % crude oil degradation, after preservation in crude oil containing medium for 40 days at room temperature, followed by switching to fresh medium containing 2% crude oil (v/v). When the culture medium was supplemented with waste frying oil (WFO) the microbial cells became highly activated, as revealed from rhamnolipid accumulation (6.8 g/l), enhanced emulsification and facilitated biodegradation of calcitrant crude oil (59 %). P. aeruginosa JW-1 was investigated to achieve the highest biodegradation percentage (77%) at initial crude oil concentration 1% (v/v). These results were confirmed by GC analysis, the profile demonstrated noticeable difference between the control and treated samples of crude oil. It could be concluded that P. aeruginosa JW-1 was able to consume most of crude oil in presence of rhamnolipid. More over, rhamnolipid production from waste product (WFO) considered economically feasible in bioremediation of hydrocarbon wastes.

 

17/29 PHYSIOLOGICAL AND ULTRASTRUCTURAL STUDY ON DICHOTOMOUS SPECIES OF CODIUM FROM THE MEDITERRANEAN SEA OF ALEXANDRIA, EGYPT (INVASION PROCESS).

Magda A. Shafik and Hala M. Taha*

Department of Biological and Geological Sciences, Faculty of Education,

Alexandria University.

*Botany Department, Faculty of Science, Alexandria University.

A group of dichotomously branched Codium species (Chlorophyta, Caulerpales, Codiaceae) were collected from Alexandria coasts. Twelve species from them were identified, classified due to invasive process to: native species represented only by one species, C. dichotomum; native in Egypt but invasion elsewhere, C. vermilara, C. tomentosum, C.taylorii, C. elongatum, C. dwarkens, C. fragile ssp. tomentosoides and C. decortiticatum; native elsewhere but invasive in Egypt, C. cylindricum and C. cuneatum and the fourth category was the alien invasive species with unknown origin (may be produced as a hybrid), C. intricatum and C. acuminatum.   Four new records were detected; two of them were alien invasive species (C. intricatum and C. acuminatum). Codium intricatum invaded Egypt at the same time of its invasion to Japan and Pillippines, then, it immigrated to Korea and Pacific. Codium acuminatum was detected in South Africa, Mozambique and Madagascar at the same time of its detection in Egypt. C. dwarkens originate in Egypt and introduced to the Pacific, Korea, Tanzania, Kenya, Pakistan, India and Australia. C. cylindricum originate in Vietnam and migrates to Egypt, Australia and other African countries. Invasion process for Codium species from Alexandria proceeded mainly from the Egyptian algal flora to other localities world-wide, little were introduced to the Egyptian marine environment. When comparing the fatty acids content of the twelve species, we find high similarity existed between C. fragile ssp. tomentosoides and C. decortiticatum, followed by C. cylindricum and C. acuminatum and between C. fragile ssp. tomentosoides and C. acuminatum as well as between C. acuminatum and C. decortiticatum. The lowest similarity was observed between C. decortiticatum and C. cuneatum. When applying the IR technique for the studied species, no coincidence occurred even between C. fragile ssp. tomentosoides and C. decortiticatum, this revealed that they are twelve different taxa. One of the most striking observations in this investigation was the presence of asexual reproductive elements (zoospores) for the two native studied algae, no sexual reproduction was detected in them; their ultrastructure also indicated that they differed from other Codium species world-wide, due to the biogeographic distribution.

 

 

18/29 SCREENING OF A HALOPHILIC ACTINOMYCETE ISOLATE FOR PRODUCTION OF SECONDARY METABOLITES BY HPLC-DAD

Wael N. Hozzein

Botany Department, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt,

Intensive programs seeking for novel natural products are running worldwide, mainly involving the screening of soil microorganisms from different habitats, which provide a rich source of natural products. In a screening program, a new halophilic actinomycete strain was isolated from a soil sample obtained from the shore of Lake Qaroun in Fayoum, Egypt to be examined for the production of secondary metabolites. The strain showed good growth in medium containing 10 to 15% (w/v) NaCl. The halophilic strain, designated H361, was identified by morphological characteristics and phylogenetic analysis and found to belong to the genus Actinopolyspora. The culture filtrate was extracted with ethyl acetate while mycelia were extracted with methanol/acetone, and then the extracts were examined by HPLC-DAD analysis for the production of secondary metabolites. The results showed that the isolated halophilic strain is biologically active, mainly producing griseolutein, saphenic acid and other unknown metabolite.

 

 

19/29 THE FIRST RECORD FOR THE INVASION OF THE RED ALGA GRATELOUPIA TO THE EGYPTIAN COASTS IN ALEXANDRIA

Magda A. Shafik and Hala M. Taha*

Department of Biological and Geological Sciences, Faculty of Education,

Alexandria University.

*Botany Department, Faculty of Science, Alexandria University

Grateloupia genus (Halymeniales, Rhodophyta) is an invasive red alga, introduced to the Mediterranean Sea in Alexandria through the last two decades. Three species were collected for the first time by the authors; G. doryphora, (Montagne), G. cutleria f. maxima (Gardiner) and G. acuminata (Holmes). Some physiological and morphological criteria were chosen as markers to differentiate between these species. Metal analysis indicated that low metal contents occurred in the three studied species; it indicated also that G. acuminata was introduced to the Mediterranean after G. doryphora and G. cutleria. The enzyme responsible for urea-degradation in G. cutleria and G. acuminata was urease, it was constitutive in G. cutleria   while it was inducible enzyme in G. acuminata. Grateloupia doryphora contained a constitutive urea-amidolyase enzyme. Active transport was the mechanism by which 14C-urea was taken up by G. doryphora and G. acuminata, on the contrary, G. cutleria showed passive mechanism. The total amino acids content and total protein amino acids of G. acuminata exceeded that of G. doryphora and G. cutleria, while the total free amino acids of G. cutleria exceeded the other two species. The scanning electron micrographs of G. doryphora and G. acuminata indicated the presence of morphological differences which may be used as a key for their identification

 

20/29 PHYSIOLOGICAL RESPONSES OF CHLORELLA SALINA

AND DUNALIELLA PARVA TO TOXICITY

OF SOME HEAVY METAL IONS

Hala M.Taha and Maysa M. Hatata

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

This work aimed to study the metabolic responses of Chlorella salina and Dunaliella parva cells to different concentrations of Zn+2, Cu+2 and Pb+2. The results cleared that best values of growth parameters of the two tested algae cultured in the basal medium (control) reached at the 8th day of culturing. Treatment of both algal species with low concentration of Zn+2 (5 ppm) stimulated growth, pigments, carbohydrates and protein contents. Higher concentrations of Zn+2 (10 and 15 ppm) inhibited the synthesis of these metabolites. On the contrary, at concentrations 5, 10 and 15 ppm of Cu+2 and Pb+2 significantly inhibited the growth and synthesis of the measured cellular metabolites in both organisms. Dunaliella was found to be more sensitive to toxicity of these heavy metals than Chlorella.

 

 

21/29 PROTEIN PROFILE OF CHROCOCCUS DISPERSUS, MICROCYSTIS FLOS-AQUAE AND MICROCOLEUS STEENSTRUQII IN RESPONSE TO CADMIUM TREATMENTS.

Ahmed D. El-Gamal

Botany and Microbiol. Dept., Fac. Sci, Al-Azhar Univ., Cairo, Egypt.

Now at College of Teachers, Holy Mecca, Biology Dept., P.O.Box 2064,

Ministry of Higher Education -Saudi Arabia

The electrophoretic studies were carried out on three selected algal species, namely Chrococcus dispersus, Microcystis flos-aquae and Microcoleus steenstruqii exposed to two different concentrations (low = 0.5 and high = 2 ppm) of cadmium. Protein profile analyzed by SDS PAGE gel electrophoresis showed differential expression of several proteins. Considerable changes in the number and the percentages of protein fractions are found for each alga. In addition, each organism reacts differently and individually in response to heavy metal exposure.

22/29 OCCURRENCE AND ULTRASTRUCTURE OF

ANAEROBIC FUNGI

Abd Al-Razak Abou Seda; Ahmed M. Ragab*; Magda A. El-Meleigy*

and Mona S. Ashour**

Botany and Microbiology Department, Faculty of science and Director of Regional Center for Mycology and Biotechnology at Al-Azhar University; *Botany and Microbiology Department, Faculty of science Al-Azhar University (Girls branch) and **Regional Center for Mycology and Biotechnology at Al-Azhar University.

The anaerobic fungi Neocallimastix frontalis and Orpinomyces joyonii were isolated from fasces of cattle and sheep. Aspergillus flavus, Aspergillus terreus and Absidia corymbifera were isolated from deep soil samples, they tolerate anaerobic stress. Morphology and ultrastructure of anaerobic and facultative anaerobic fungal isolates were carried out using scanning and transmission electron microscopy. Morphology and ultrastructure of facultative anaerobic fungal isolates were metamorphosed under anaerobic stress.

 

23/29 ENHANCEMENT OF HYGROMYCIN A PRODUCTION BY STREPTOMYCES HYGROSCOPICUS NRRL 2388 USING MUTAGENESIS AND A MIXTURE OF INOCULA

El-Sayed E. Habib and Mohamed A. El-Mowafy

Microbiology Department, Faculty of Pharmacy, Mansoura University, Egypt

Hygromycin A, an antibiotic produced by Streptomyces hygroscopicus NRRL 2388, is an inhibitor of bacterial ribosomal peptidyl transferase. The antibiotic is effective against multiple resistant pathogens, especially against Serpulina (treponema) hyodysenteriae, the causative agent of animal dysentery. Reported herein are the results of trials to enhance the production of hygromycin A using mutation and a mixture of inocula. A maximum production of hygromycin A (1860 mg/ml) was observed when UV and 5-Flurouracil (5 FU) mutant strain was used after 5 days incubation, compared with UV mutant strain (1630 mg/ml) after 6 days incubation, and its wild strain (1450 mg/ml) after 6 days incubation. The most effective mixing ratio of inocula for hygromycin A production (1990 mg/ml) in shake flasks was 3:1 volume ratio of early- and late-stage inocula, at initial pH 7.0, and 30oC after 5 days of incubation and agitation at 150 rpm. In the fermenter studies using 2 liters capacity Braun fermenter, the production of hygromycin A using the highly producing mutant strain (UV & 5 FU mutant strain and 3:1 volume ratio of early and late-stage inocula) was decreased by 25% compared to the amount produced by shake flask using the same mutant strain.


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