Vol. 18, October, 2004

cost of viagra in quebec 1/18 AMBROXOL INDUCTION OF MULTIPLE DRUG RESISTANCE IN PSEUDOMONAS AERUGINOSA AND ENTEROBACTER CLOACAE

   M.A.M. Yassien

Department of Microbiology, Faculty of Pharmacy, King Abdulaziz University,

Jeddah, Saudi Arabia

The growth of comment avoir viagra sans ordonnance Pseudomonas aeruginosa and pilule style viagra Enterobacter cloacae in 30 ug/ml ambroxol showed development of multiple-drug resistance to ciprofloxacin, norfloxacin, ofloxacin, cephalothin, cefoxitin, ceftazidime, cefotaxime, and aztreonam, but, no change in the susceptibility of the tested strains to amikacin was observed . The MICs values of the used fluoroquinolones, β-lacatams, and monolactams against the tested strains were increased in the presence of ambroxol by 8-16 , 16-32, and 8-16 times, respectively, as compared to the control (without ambroxol). The developed resistance was transient and nonheritable. The presence of ambroxol showed a significant reduction (3- to 6-fold) in the norfloxacin uptake and inhibition in the synthesis of the 40KDa and 41KDa outer membrane proteins in prendre du viagra apres un infarctus Pseudomonas aeruginosa and cialis courbatures Enterobacter cloacae, respectively, was observed. On the other hand, the presence of ambroxol did not affect the microbial production of β-lactamase and the susceptibility of DNA gyrase, extracted from the tested strains, to norfloxacin. These results indicated that ambroxol induced a phenotypic multiple drug resistance in prix vente viagra pharmacie Pseudomonas aeruginosa and ou commander cialis en france Enterobacter cloacae through suppression the synthesis of 40KDa and 41KDa outer membrane proteins, respectively, which play a role in the antimicrobial permeation.

 

 

2/18 DEGRADATION OF AROMATIC COMPOUNDS BY IMMOBILIZED ACINETOBACTER JOHNSONII

FROM SAUDI-ARABIA

S.R. Salem and F.A. Al-Barakati*

Faculty of Education, Alexandria University, Egypt

*Faculty of Education, Ministry of Education, Makkah Al-Mukkarramah,

Saudi- Arabia

All the bacteria isolated from Makkah Al-Mukkarramah region could grow on solid culture of minimal medium containing 0.1g/l of different aromatic compounds such as phenol, m-cresol, p-cresol, naphthalene and anthracine and couldn’t grow on 0.5g/l except in case of phenol. durĂ©e de l'effet du cialis Acinetobacter johnsonii was the best strain to grow on phenolic compounds at different rates, it showed a high rate of degradation on phenol (41.67 mg/l/h), and lower rates on m-cresol (19.23 mg/l/h), p-cresol (17.86 mg/l/h), 4-chlorophenol (10.42 mg/l/h) and bromophenol (8.93 mg/l/h). Cells of viagra pour femme puissant A. johnsonii were immobilized by entrapment in agar- alginate gel and proved to be superior to free cells in phenol degradation by 2-fold while adsorbed cells on sponge showed 1.7- fold degradation rate of that of free cells. The rate of degradation by using an agar gel concentration of 2% was the best concentration where attained 1.5, 1.7 and 1.8 fold, respectively. A bead volume of 20 ml / 100 ml culture medium achieved a maximum level of degradation and least value of cell leakage. Reincubation of the entrapped cells increased the rate of phenol degradation by 1.5-fold.

kamagra aphrodisiaque naturel 3/18 GENERATION OF GENETICALLY ENGINEERED BANANA EXPRESSING b-GLUCURINIDASE VIA AGROBACTERIUM-MEDIATED GENE TRANSFER AND MICROPROJECTILE BOMBARDMENT

S.M. Khalil, M.E. Wagih* and E.E. Wagih**

Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt

*Biotechnology Centre, University of Technology, Lae, Papua Niugini, North of Australia

**Department of Plant Pathology, College of Agriculture, University of Alexandria, Alexandria, Egypt.

Genetically engineered banana plants expressing β-glucuronidase (GUS) were produced using particle gun-wounded apical meristem and Agrobacterium tumefaciens strain EHA105 harboring the pBI121 plasmid carrying NPT-II and GUS genes. Longitudinally bi-sectioned apical meristem was bombarded (at 650 psi) with gold particles (1 μm diameter) coated with plasmids containing the neomycin phosphotransferase (nptII) selectable marker gene under the control of the NOS promoter and the b-glucuronidase (uidA) reporter gene under the control of the CaMV-35S promoter. Alternatively, longitudinally bi-sectioned apical meristem was bombarded at the same psi with naked gold particles of the same diameter and co-cultivated for 30min with Agrobacterium. After three days from co-cultivation with the bacterium, the apical meristem was subjected to selection on a regeneration medium supplemented with 110 mg/dm3 kanamycin and 500 mg/dm3 carbincillin. Successful integration of genes was confirmed by PCR and southern blotting, while expression of uidA gene was hestochemically demonstrated in different parts including meristem, true leaves and roots of the transgenic banana; confirming both integration and high expression of uidA gene in the transgenic banana plantsobtained.

4/18 ISOLATION, PHYSICOCHEMICAL AND IMMUNOLOGICAL CHARACTERIZATION OF GALECTIN-1 FROM SHEEP BRAIN

M. Shahwan, T.M. Al-Qirim, M.S. Mansy*, S.M.K.R. Zaidi** and N. Banu**

Department of Biochemistry, Faculty of Pharmacy, Al-Zaytoonah University of Jordan, Amman, Jordan

*Department of Microbiology and Immunology, Faculty of Pharmacy, Al-Zaytoonah University of Jordan, Amman, Jordan

**Department of Biochemistry, Faculty of Life Sciences, A. M. University Aligarh-U.P. India

The affinity purified galectin moved essentially as a single polypeptide band of 14.1 kDa on SDS-PAGE. The gel filtration under reducing and non reducing conditions revealed its molecular weight as 14.3 and 28.5kDa respectively, suggesting that the sheep brain galectin is a homodimer. The most potent saccharide inhibitors tested were lactose, galactose. D-b-galactosamine, methyl-b-D-galactopyranoside. The exposure of sheep galectin to oxidizing agent destroyed its hemagglutination activity, and shifted its UV absorption maxima from 280 to 250 nm. Loss of galectin activity was observed after treatment with iodoacetate, iodoacetamide, pHMB and NEM. The galectin exhibited preference for trypsinized human type A erythrocyte rather than O, B and AB.The antibodies raised against the pure lectin showed immunological relationship between sheep brain galectin and other brain galectins of various vertebrate origins.


5/18 ISOLATION, MOLECULAR CLONING AND SEQUENCING OF COAT PROTEIN GENE OF CUCUMBER MOSAIC CUCUMOVIRUS FROM EGYPT

S.M. Khalil, M.E. Wagih* and E.E. Wagih**

Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt

*Biotechnology Centre, University of Technology, Lae, Papua Niugini, North of Australia

**Dept. of Plant Pathology, College of Agriculture, University of Alexandria, Alex. Egypt,

The coat protein gene of a cucumber mosaic cucumovirus (CMV-CP) isolate infecting banana and squash plants was isolated, cloned and sequenced. A reverse transcription-polymerase chain reaction (RT-PCR) technique was developed for the amplification of CP gene from viral RNA. The RT-PCR was conducted using two specific oligonucleotides to CMV-CP. The CP gene was confirmed by nested–PCR using internal primers and Southern blot hybridization using 32P-labelled probe. The RT-PCR product (681 bp) was labelled and used as a probe to detect CP gene in recombinant E. coli. JM109.   Complete nucleotide sequence and the deduced amino acid sequence of the CP gene revealed an open reading frame coding for 218 amino acids. Based on nucleotide and amino acid sequences, the CMV-CP of the Egyptian isolate was found to share 90 to 96% nucleotide sequence and 93 to 96 amino acid sequence with those of eight other isolates listed in the GenBank.

6/18 BIOCONVERSION OF HEMICELLULOSES, BLACK LIQUOR WHICH POLLUTED WATER INTO ORGANIC ACIDS

S.S. Abd-El-Salam

Botany Department, Faculty of Science Benha, Zagazig University

The studies on the conversion of waste products have shown a wide potential by microorganisms to produce beneficial compounds from waste products. The hemicelluloses of black liquor of rice hulls was used after separation of lignin and silica contents. Hemicelluloses constituted from hexoses and pentoses where the major part was hexoses while pentoses was the minor part, the hexose composition was mannose, rhamnose, galactose and glucose while the pentose content of the black liquor consisted of arabinose and xylose. By using several strains of bacteria which were isolated from acidified fermented black liquor basal medium with acetic acid at pH 6.5. These isolated fragments were identified as acetic acid bacteria by classical scheme and using finger print of DNA. These strains are able to grow and utilize carbohydrates of black liquor to convert it to organic acids via oxidation producing gluconic, citric, succinic, malic, fumaric and acetic acid which were identified by using HPLC system at 214 nm., these acids are useful products with an industrial importance.

7/18 EFFECT OF GAMMA IRRADIATION AND PHOSPHORUS ON THE COMPOSITION OF CHAMOMILE ESSENTIAL OIL PRODUCED IN VITRO AND IN VIVO

A.H. Nassar, M.F. Hashim*, N.S. Hassan** and H. Abo-Zaid*

Department of Botany, Faculty of Science, Ain Shams University, Cairo, Egypt.

* Nuclear Energy Authority, Inshas, Egypt.

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

The effect of different doses of gamma irradiation and phosphorus on the composition of chamomile essential oil produced in flower heads or callus cultures was studied. Seeds of chamomile were irradiated with 0, 2, 6 or 8 K-rad Gamma rays and grown in a soil provided with phosphorus in the form of calcium superphosphate (20 Kg/Fed.) to full flowering stage. Irradiated seeds were also used to establish callus culture line using Murashige and Skoog medium supplemented with2.0 mg/L kinetin, 0.5 mg/L 2,4-dichlorophenoxy acetic acid, and 300 mg/L phosphorus (as dihydrogen potassium phosphate). Essential oil was extracted from chamomile flower heads and callus using “Supercritical Fluid Extraction” method with CO2.Eight components in the flower head oil and seven components in the callus oil were identified. Bisabolol oxide A was found to be the major component in both in vivo and in vitro oil. Qualitative similarity was observed between both oils. However, quantitatively the amount of oil produced in callus was much less than that produced in the flower heads. Low dose of gamma irradiation (2 K-rad) enhanced the production of some components on the expense of others. Moreover, the percentage of matricine (chamazulene) production in callus tissues was higher than that of the flower head.

8/18 INCREASING THERMOTOLERANCE OF PISUM SATIVUM L. PLANTS THROUGH APPLICATION OF PUTRESCINE AND STIGMASTEROL

H.M.S. El-Bassiouny

Botany Department, National Research Centre, Cairo, Egypt.

     Two pot experiments were carried out during two successive seasons of 2002 & 2003 and sowing at two dates September 1st and November 1st to study the effect of early sowing date at high temperature and presoaking in putrescine (10-5, 10-4 & 10-3 M) and stigmasterol (10-8, 10-6 &10-4 M)   on growth, yield components,   endogenous phytohormone, photosynthetic pigments, proline accumulation and protein profile of Pisum sativum plant. The early date of sowing at high temperature ranged between (38 – 32°C) in general caused marked reduction in shoot length, number of branches and leaves per plant, fresh and dry weights of shoots, number of pods per plant, number of seeds per pod and weight of seeds per plant. The high temperature also showed marked decreases in endogenous promoters (i.e IAA, GA3 and cytokinins) and photosynthetic pigment contents concomitantly with increases in ABA and proline contents. Soaking of Pisum sativum seeds in putrescine or stigmasterol at the selected two dates significantly increased growth and yield components concomitantly with increases in the amounts of IAA, GA3, cytokinins, photosynthetic pigments and proline and decreases in the amount of ABA. The changes in protein pattern in Pisum sativum L shoots at suitable (26-23oC) and high temperatures (38—32oC) and treatment with putrescine and stigmasterol were investigated. High temperature induced the synthesis of new proteins these proteins are members of heat shock proteins. The induced heat shock proteins have molecular weights (104, 90, 70, 36, 29 and 23 K Da.) which were apparent in Pisum sativum L shoots. The putrescine application induced synthesis of certain responsive proteins (Mwt. 94 & 41 K Da.), while the stigmasterol application induced the synthesis of the responsive proteins (M wts 110 & 41 K Da). Putrescine and stigmasterol mostly increased the intensity of some HSPs (90, 70, 36, 29 & 23 KDa.). These proteins might interfere with increasing thermotolerance of Pisum sativum plants.

9/18 EFFICIENCY OF BIOFERTILIZERS IN IMPROVING DROUGHT STRESS TOLERANCE IN BARLEY

H.M. Emad El-Din, M.H. Rashad* and E.A. Ismail**

Botany Dept., Fac. of Science., *Plant Physiology Section, Botany Dept., Fac. of Agric., **Agronomy Dept., Fac. of Agric Cairo Univ. Giza, Egypt.

Two pot-experiments were conducted during 1998/99 and 1999/2000 seasons to examine the efficiency of biofertilizer as a substitute for chemical fertilizers as means of improving drought stress tolerance in barley. Biofertilizer-inoculated (with ½ NPK dose) and non-inoculated (with full NPK dose) treatments were applied to five barley genotypes (Giza 123, Giza 125, Rehan 97, Morocco and a wild genotype) under three moisture levels (100, 70 and 40%) of field capacity (FC) in five replications. Chemical constituents, vegetative and yield attributes as well as proline and chlorophyll contents were estimated. Significant differences were found between the three levels of moisture in both vegetative and yield attributes. There was no significant difference between biofertilizer-inoculated (with ½ NPK dose) and non-inoculated control (with full NPK dose) in both vegetative or yield attributes. This indicated that biofertilizers could be efficient in reducing chemical fertilizers under drought stress. Also, significant differences were found among the genotypes in most studied attributes, where Giza 123 and Rehan 97 had relatively high values for most vegetative and yield attributes as well as chemical constituents, which refers to differential response of barley genotypes under drought stress and biofertilizer inoculation.

10/18 CHARACTERIZATION OF TOBACCO MOSAIC VIRUS-EGYPTIAN STRAIN: CLONING AND SEQUENCING OF 3'-TERMINAL GENOME SEQUENCE

S.A. Shoman

Department of Microbiology, Faculty of Science, Ain Shams University,

This investigation deals with the molecular characterization of the TMV isolated from tomato (Lycoporsicon esculentum) in Egypt. Both TMV-RNA genome as well as its coat protein were extracted and analyzed from the purified virus preparation. The genomic sequences of the 3'-terminus encompassed the coding regions of coat protein and 3'-untranslated region (UTR) of approximately 689 nucleotides were selected and amplified with synthetic oligonucleotide primers of tobamovirus using reverse-transcription polymerase chain reaction (RT-PCR) to confirm the identity of the virus. The amplified PCR product was cloned into a pGEM-T easy vector subjecting to sequence in the both directions. These sequence data of TMV Egyptian strain was compared with the genomic sequence of other TMV-strains according to the data in GenBank. Sequence analysis showed 95% identity of nucleotide sequence, while the deduced amino acid sequence of the coat protein showed 86 % of similarity and marked amino acid changes at C-terminal region. These data confirmed that the virus under study is an Egyptian strain of TMV which is referred to herein as TMV-E.  

11/18 EFFECT OF APPLICATION OF SOME UREA DERIVATIVES ON GROWTH AND BIOCHEMICAL ACTIVITIES OF PLEUROTUS OSTREATUS

A.S.M. Mousa

Botany Department, Faculty of Science, Beni-Suef Branch, Cairo University

The effect of urea and urea derivatives (urea nitrate, urea sulphate, and urea phosphate) and their combinations as nitrogen sources were studied on Pleurotus ostreatus mushroom grown on MCM agar medium. The maximum growth was obtained with the mixture of urea nitrate +urea sulphate+ urea phosphate. Then, P. ostreatus was cultivated on rice straw treated with this urea mixture. There was an increase in the yield of fruiting bodies and a decrease in the culture cycle. The biochemical composition of mushroom showed an improvement in all estimated parameters (carbohydrate fractions, nitrogen fractions, amino acids, fatty acids and lipids, nucleic acids and mineral contents). These obtained results may suggest that, this treatment can be applied to obtain mushroom with a good value of protein and minerals for humans.

12/18 INHIBITORY EFFECT OF POLYGONUM SALICIFOLIUM L. EXTRACT ON SOME PLANT PATHOGENIC FUNGI

S.E.A. Khodary, A.S.M. Mousa, K.H. Shaker* and H.T.M.I. Sweelam

Botany Department, Faculty of Science, Beni-Suef Branch, Cairo University

*Department of Chemistry of Natural and Microbial Products, National Research Center, Cairo, Egypt.

Fusarium oxysporum was isolated from wilted tomato roots while Alternaria alternata and Penicillium digitatum were isolated from rotten tomato and orange fruits, respectively. The methanol extract and its fractions of the aerial parts of Polygonum salicifolium L. (Polygonaceae) were tested for their antifungal activities against the isolated pathogens (A. alternata, F. oxysporum and P. digitatum). Ether-ethyl acetate fraction of P. salicifoliumsuppressed spore germination, dry weight and mycelial growth of the tested fungi. The fraction was highly effective for control of Fusarium wilt of tomato seedlings and for control of Alternaria and Penicillium rots of tomato and orange fruits respectively. Three flavonoids were isolated from the active fraction of P. salicifolium using column chromatography and were identified as quercetin, quercetin 3-O-glucoside (isoquercitrin) and quercetin3-O-galactoside (hyperin). The MICs of these compounds were 950 µg/ml for quercetin and 700 µg/ml for the mixture of quercetin-3-O-glucoside and quercetin-3-O-galactoside.

13/18 SOIL ALGAE IN DIFFERENT HABITATS AT EL FAYOUM GOVERNORATE (EGYPT).

M.H.M. Abdel-Rahman, R.M.Ali and H.A.Said

Botany Department, Faculty of Science, Cairo University, El Fayoum, Egypt.

The investigated area extends about 25-30 Kilometers to the north of El Fayoum city (at about 100 km south west of Cairo, Egypt). Three sites (I, II and III) were chosen mainly on the basis of their differences in salinity (I < II < III) and soil texture. The algal biomass and distribution could be mainly correlated to the alterations in soil type. The pH values of the investigated soil samples were generally of the alkaline side during the study period. During the whole investigation, the maximum value of organic carbon was recorded at site III in spring and autumn while minimum value at site I in winter. The exchangeable cations in the investigated soil samples had the following order Ca+2 > Mg+2 > K+ > Na+. The three most abundant groups of soil algae at all the sites surveyed were Chlorophyta, Bacillariophyta and Cyanophyta. Euglenophyta were recorded only in the soil samples collected from site III during all the seasons. Twenty eight genera (42 species) of algae were isolated from the three various soil samples throughout the investigation period out of these: seven genera (10 species) belong to Chlorophyta, eleven genera (14 species) to Cyanophyta, eight genera (16 species) to Bacillariophyta and two genera (2 species) to Euglenophyta. Among the predominate genera of green algae throughout the investigation period were Chlorella vulgaris and Chlorococcum humicola.

14/18 PRODUCTION, CHARACTERIZATION AND IMMOBILIZATION OF LIPASE OF RHIZOPUS OLIGOSPORUS NRRL 2549 VIA SOLID STATE FERMENTATION USING MUSTARD SEEDS AS SUBSTRATE

M.E. Moharam and S.T. El-Sayed

Microbial Chemistry Department and Biochemistry Department, National Research Center, Dokki, Cairo, Egypt.

Rhizopus oligosporus NRRL 2549 was selected from a screening programmed for lipase production via solid state fermentation conditions (SSF). Lipase production by Rhizopus oligosporus NRRL 2549 using milled mustard seeds as a substrate gave maximum activity among all tested substrates. The optimized medium and cultural conditions consisted of milled mustard     seeds supplemented with 1% peptone and added moisture 30% (w/w) in the presence of 12:1 air :medium ration per conical flask after 3 days of incubation at 28 C under SSF conditions. Optimum temperature and pH for enzyme   activity were 37 C and pH 3 respectively. The enzyme showed hydrolytic activity against corn, olive and sun flower oils. R. oligosporus NRRL 2549 lipase was immobilized by simple adsorption on Tricalcium phosphate (TCP) gel and egg shell powder with specific activity of 18.3 unit /mg protein and 13. 1 unit /mg protein respectivily compared with   0.62 unit /mg prrotein for free enzyme.The optimum TCP concentration was 40:1 adsorbent: enzyme protein .Some properties of free and immobilized enzyme were studied. Optimum reaction temperature and pH of immobilized enzyme were 40 C and pH 3. Substrate specificity of immobilized enzyme revealed preferable specificity towards corn oil followed by olive and sun flower oils.

 

 

15/18 BIOCHEMICAL MEANS FOR INCREASING THE POTENCY OF BACILLUS SPHAERICUS MOSQUITOCIDAL ACTIVITY AGAINST CULEX PIPIENS

M.A. El-Bendary, M.E. Moharram and M.S. Foda
Microbial Chemistry Department, National Research Center, Cairo, Egypt
         Simple biochemical means were used to enhance the activity of Bacillus sphaericus (Bs) mosquitocidal toxins through optimization of the conditions present in the larval gut of Culex pipiens (target insect species). This goal was achieved by incorporation of some chemical compounds that could act synergistically with Bs toxins without exerting any harmfull effects on human and environment. Calcium chloride (0.07%), potassium carbonate (0.25%), and ammonium phosphate (0.25%) increased the activity of the Bs toxins in varying degrees (2.5, 1.3 and 4 folds respectively). Among the common and safe organic acids and their salts, succinic acid (0.1%), ammonium oxalate (0.5%) and acetamide (1%) showed a potentiation in Bs mosquitocidal activity against Culex pipiens leading to decrement of LC50 of Bs toxins about 2.4, 2.2 and 1.7 fold respectively. Some protein solubilizing agents, sodium thioglycolate (0.1%), dipotassium hydrogen phosphate (0.5%) and EDTA (0.025%) were also potentiative chemicals for the mosquitocidal toxins through lowering the LC50 of Bs toxins about 1.8, 3.2 and 3.3 folds respectively. On the other hand, lipid emulsifying agents tested showed additive or antagonistic effect for Bs toxin. However, vegetable oils and chemical insecticides had antagonistic effect for Bs toxin under study. It is of interest to note that the tested vegetable oils exhibited mosquitocidal activity when tested as such at low concentration. Neem oil is the most active one with LC50 at 0.01%, whereas 100% larval mortality was obtained at 0.02% final concentration. The possible application of this approach in the commercial production of bacterial insecticides through incorporation of these chemical additives at a post harvest stage, will be of great value and more economic.

16/18 RAPID DETERMINATION OF TOTAL LIPIDS FROM MILK AND MILK PRODUCTS BY SUPERCRITICAL CARBON DIOXIDE

S.A. El-Behairy

*Department of Food Evaluation and Food Science, National Organization for Drug Control and Research (NODCAR), Giza, Egypt

Total lipids of milk and milk products (0.9-36 % fat) were extracted by supercritical carbon dioxide (SC-CO2). The extraction was carried out at 60 °C over a pressure range of 300-500 bar with using ethanol as a modifier. Percent recovery of total lipids obtained by SC-CO2 was 99-105 % as compared with Rose-Gottlieb method. Adding ammonia (1 μL/10 mL) prior to extraction did not affect the recovery. At any pressure, increasing the extraction time up to 40 min conduced to a considerable increase in the recovery. Differences in recovery with Modifier ratio (1 and 5 %) were more obvious at higher pressures and lower temperatures.

17/18 DETECTION AND SOME CHARACTERISTICS OF BEET NECROTIC YELLOW VEIN BENYVIRUS FROM RHIZOMANIA-AFFECTED SUGAR BEET IN EGYPT

M.H. Abdel-Ghaffar and E.S.H. Farrag*

Department of Agricultural Microbiology (Virology Lab.), Faculty of Agriculture, Ain Shams University, Cairo, Egypt.

* Department of Plant Pathology, National Research Center, Dokki, Giza, Egypt.

In this study, the transmission experiments indicated that beet necrotic yellow vein virus (BNYVV) was mechanically transmitted to Chenopodium amaranticolor, Chenopodium quinoa and Beta macrocarpa inducing chlorotic local lesions, which spread into the veins.Virus also produced systemic infection on Sugar beet (Beta vulgaris cv. Pleno) and Spinacia oleracea. Indirect DAS-ELISA usingpolyclonal antibodies to the C-terminal 60 amino acids of the BNYVV coat protein was also done to confirm the diagnoses based on test-plant reactions The results of fungal transmission showed that the BNYVV was also transmitted through the cystosori of Polymyxa betae-inoculated soil. The results were serologically confirmed by the positive reactions on indicator host, Indirect DAS-ELISA and the light microscope revealed the presence of P. betae cystosori inside the cells of lateral root of B. macrocarpa plants. The virus was purified from BNYVV-infected B. macrocarpa tissues, and the obtained yield was about 1-1.2 mg/100 g infected tissue. The results of examination the purified virus preparations by transmission electron microscope indicated presence rigid, rod-shaped virus particles with a central core. The virus particles were 20 nm wide and ranged from 50-400 nm in length. BNYVV coat protein was migrated as single band with molecular weight of about 21 kDa, estimated in 12% SDS-PAGE. The BNYVVnucleic acid extracted from purified virus preparation and electrophoresed through 1% agarose gel under denaturing conditions was separated into four discrete bands with estimated sizes of about 6.7, 4.9, 1.8 and 1.4 kb. The obtained results confirmed the specificity of the synthesized primers to coat protein gene encoded by BNYVV RNA-2 with the expected products of about 550 bp which was amplified from RT-PCR reactions in presence template nucleic acids extracted from purified virus or infected tissue.

18/18 THE INTERACTIVE EFFECTS OF NACL AND ASCORBIC ACID OR KNO3 ON GROWTH AND SOME RELATED PHYSIOLOGICAL ACTIVITIES OF CHLORELLA VULGARIS AND CHLOROCOCCUM HUMICOLA.

M.H.M. Abdel-Rahman, R.M. Ali and H.A. Said

Botany Department, Faculty of Science, Cairo University, El Fayoum, Egypt.

The interactive effects of salinity (NaCl) and ascorbic acid on the growth and physiological activities of Chlorella vulgaris Beij. and Chlorococcum humicola (Näg.) Rab. were investigated in this work. Low and moderate salinizations stimulated the growth of Chlorella and Chlorococcum, in the latter alga growth was reduced by high salinity levels. The application of ascorbic acid or KNO3 led to a significant increase in the values of growth parameters and biosynthesis of pigments at low and moderate stressed algal cells and the adverse effects of high levels of salinity, in some cases, were partially alleviated when compared to the reference control. Ascorbic acid in most cases was more effective in alleviating the stress effects of salinization than KNO3. Considerable decrease in fractions and total contents of carbohydrates and proteins was induced by salinity stress in both tested organisms. Ascorbic acid or KNO3 treatments of salinized cells of both algae exhibited a general increase in their contents of carbohydrates and proteins (total and fractions). Increasing NaCl level forced the tested organisms to synthesis proline and other free amino acids. However, treatments of both algae with ascorbic acid or KNO3 considerably lowered the contents of proline and free amino acids, a response which was, generally, accompanied by the biosynthesis of proteins and the inhibition of their dissimilation. It was concluded that treatment with ascorbic acid or KNO3 partially or completely alleviate the adverse effects of salinity on growth and physiological activities of both studied algae.

19/18 BIODEGRADATION OF HIGH MOLECULAR WEIGHT AROMATIC HYDROCARBONS BY FUNGAL-BACTERIAL CONSORTIUM.

R.A. Bayoumi

Botany & Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, PN: 1884, Egypt.

This study investigated the biodegradation of anthracene and phenanthrene in liquid media by Penicillium chrysogenum strain APH2 and Pseudomonas aeruginosa strain APH2 those were isolated from heavy oil polluted soil samples. Both polycyclic aromatic hydrocarbons (PAHs) compounds were utilized by both most potent isolates as sole sources of carbon and energy. Culture of Penicillium chrysogenum APH2 strain was dosed with anthracene or phenanthrene, and after 21 days of incubation of the added anthracene and phenanthrene had degraded. The metabolites were extract, and identified by Gas Chromatography-Mass Spectrometry (GC-MS) and compared with to WILEY Mass Spectral Database (Searched library). The chloroform extracted from anthracene incubated with Penicillium chrysogenum for 14 days showed six metabolites, identified as Benzyl ester of benzoic acid, phenanthrene, 9H-Carbazole, 9,10-Anthracenedione, 9(10H)-Anthacenone, and Benzene, 1-Chloro-3-(Phenylethynyl). Phenanthrene ring fission products, isolated from chloroform extract after incubation at 30°C for 14 days with Penicillium chrysogenum APH2 strain were identified as 9H-Fluorene, Benzene, 1,1-(1,2-ethenediyl)bis, 9a,10-Dihydrobenz [a] azulene, Benzyl ester of benzoic acid, Dibenzothiophene, Anthracene, Anthraquinone, 9-Chloro-9,10-Dihydro-9-Boranthracene, and 1,2,4-Benzene tricarboxylic acid,4-dodecmm dimethyl ester. The chloroform extract from anthracene was incubated for 21 days with mixture of Penicillium chrysogenum–APH2, and Pseudomonas aeruginosa-APH2 and showed eight metabolites were identified as Benzene, 1,1-(1,2-ethenediyl)bis, dibenzothiophene phenanthrene, 9,10-Anthracenedione, Tricosane, Eicosane, Docosane, and Heneicosane. Phenanthrene ring fission products isolated from chloroform extracts after incubation with mixture of Penicillium chrysogenum–APH2, and Pseudomonas aeruginosa-APH2 at 30°C for 21 days showed five metabolites were identified as 2-bromoethyl benzene, anthracene, nonadecane, heptacosane, and tetradecane. Application of a consortium metabolized height molecular weight hydrocarbons anthracene or phenanthrene to long chain aliphatic hydrocarbons which are easy to be biodegraded by any microorganism.

20/18 CHANGES INDUCED BY IAA AND COUMARIN IN SEED GERMINATION AND IN SOME METABOLITES OF RADISH SEEDLINGS

W.A. Kasim, A.A. Mohsen, S.M. Ebrahim* and A.N. El-Sebaie

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

*Department of Pharmacognosy, Faculty of Pharmacy, Tanta University,

Tanta, Egypt.

Seeds of Raphanus sativus L. aegyptiaca were germinated either in distilled water, 10-5 M IAA, 10 –3 M coumarin or in IAA + coumarin for 6 days. The effects of these treatments on the growth criteria, photosynthetic pigments (Chl. a, Chl. b, carotenoids), nucleic acids, activity of the two transaminases GOT and GPT, carbohydrates and different nitrogenous components were recorded.The results seem to indicate that: (i) the radicle is more sensitive to the effects of IAA, while the shoot is more sensitive to the impact of coumarin, (ii) the suppressive effect of the different treatments on the fresh weight and succulence of the seedlings may well be taken as an indication oftheir inhibitory effect on water uptake, (iii) the activity of the photosynthetic pigments is not a function of their content in the cotyledonary leaves but rather of their efficiency, (iv) IAA and coumarin have clearly contradictory impacts on the carbohydrate contents of the seedlings and their separate organs, as IAA increased their total soluble carbohydrates and total carbohydrates but decreased their starch content while the opposite was the case with coumarin, (v) the remarkable increase of the soluble sugars as a result of IAA treatment, accompanied by reduction in the starch content could be ascribed to a stimulatory effect of the phytohormone on starch degradation and/or an inhibitory effect on starch biosynthesis, (vi) the decrease in GOT and GPT activities might be attributed to disturbances in RNA and DNA systems, and (vii) IAA and coumarin have contradictory effects on the insoluble nitrogen and the total nitrogen contents of the seedlings, but when in mixture they have a highly restorative impact on both of their soluble and insoluble nitrogen components.

21/18 STUDIES ON ACINETOBACTER BUMANNII ISOLATED FROM INTENSIVE CARE UNITS

H.K. Abd El-Latif and *H.K. Abd El-Latif

Department of Microbiology, Faculty of Pharmacy, Zagazig University

*Department of Anesthesiology, Faculty of Medicine, Zagazig University

Acinetobacter spp are opportunistic pathogens, commonly associated with hospital acquired infections. In these studies, Acinetobacter spp. were collected from intensive care units (ICUs) from Al-Riyadh and Makka cites, Saudia Arabia. Acinetobacter baumannii was the most frequently species among genus Acinetobacter. The antimicrobial susceptibility patterns were determined for the clinical isolates of A.baumannii. All clinical isolates of A.baumannii exhibited multiresistance profile to most of the tested antimicrobial agents including b-lactam drug or non b-lactam drug. In our study, all isolates of A.baumonnii were sensitive to imipenem. Also, reference organic A.calcoaceticus was multiresistant organism. The obtained results revealed that no difference in the MICs for penicillin G, gentamicin and ciprofloxacin were found with the presence or absence of reserpine suggesting that efflux mechanism was not present in the tested organism. Also, outer membrane protein (OMP) profile of the clinical isolates and reference organism detected that both of two types of organisms had nearly the same OMP profile, while the bands of A.calcoaceticus was more thiner and faint than bands of corresponding proteins in clinical isolates, these may indicated the overproduction of OMPs in clinical isolates which may be responsible for resistance. The plasmid analysis showed that most of clinical isolates of A.baumannii had the same plasmid pattern. The resistant isolates showed one or two plasmids DNA of size about 140, 12, 10 MDa. Some clinical isolates were free from any plasmid. The DNA plasmids of clinical isolates were transformed to E.coli competent cell C600 isolate number 103, 104, 105 has very low rate of transformation (60 CFU/20 mg DNA) no transformation occurs with other isolates these data revealed that resistance may be carried on transposons or integran as reported previously.  

22/18 SYSTEMATIC AND MICROMORPHOLOGICAL STUDIES ON THE GENUS AMARANTHUS L. IN EGYPT

M.M Zareh and N. El-Husseini*

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

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

A systematic revision for the species of Amaranthus L. was provided. Twelve species with 2 subspecies and four varieties are recorded. Leaf surface and seed testa were studied using light and scanning electron microscopy. Three leaf surface types and four seed microsculpturing patterns were recognized; description of each type and pattern, key to the studied species as well as SEM micrographs of leaf surface and seed microsculpture were provided.

23/18 BIOLOGICAL AND BIOCHEMICAL STUDIES ON SOME STREPTOMYCES SPECIES LOCALLY ISOLATED

1-      ISOLATION, TAXONOMICAL IDENTIFICATION AND ANTIFUNGAL POTENTIALITIES

M.R. Abu Shady*; F.M. Elbeih*; A.A. Elgammal; A.L. Kansoh and A.A. Keera

*Botany Dept. Fac. Sci. Ain Shams Univ. and Microbial Chem. Dept. National Research Centre (NRC), Cairo – Egypt.

Various Streptomyces isolates obtained from different soil regions of Egypt were taxonomically identified. This collection included forty two isolates belonging to the grey, rose and white series of streptomycetes. The isolates of each series of Streptomyces were differentiated according to the morphological characteristics into three sections: Section A with straight spore chains includes 9 isolates, section B with hooked spore chains includes 10 isolates and section C with spiral spore chains includes 23 isolates. all those isolates were identified according to the cultural, physiological and antagonistic characteristics. An organism from each section in the grey, rose and white series was chosen as a representative example in this work. The antifungal potentialities of the studied isolates were determined against different moulds and yeasts.

24/18 CHANGES IN GROWTH, PROTEIN PATTERNS, DNA FINGER PRINTS AND CHROMOSOMAL ABERRATIONS OF WATER STRESSED MAIZE SEEDLINGS TREATED WITH ABSCISIC ACID OR JASMONIC ACID

S.M. El-Khallal and Th.R. Mohamed

Botany Department, Girls College for Arts, Science and Education.

Ain Shams University, Cairo, Egypt.

Abscisic acid (ABA) and jasmonic acid (JA) have been implicated in responses to water stress. External application of ABA (50μM) and JA   (5μM) as seed treatment before subjected to water stress with polyethylene glycol 6000 (PEG, 0%, 20% and 40 %) for 48h can alleviate the harmful effect of water deficit and increase protection of plant cells against dehydration. Thus, ABA and JA- treated plants had a higher growth rate (length, fresh and dry weights of shoots and roots), nucleic acids and soluble protein levels, as compared with the un-treated ones. In an attempt to elucidate the possible roles of ABA and JA in the mechanism of drought resistance, changes in gene expression and protein profile in stressed maize seedlings were investigated. PCR – RAPD analysis of genomic DNA isolated from stressed seedlings revealed the occurence of 10 distinct DNA fragments. Three of them which having 9116, 1035 and 906 pbs could be considered as specific for DNA isolated from ABA and JA treated plants and exposed to 40% PEG. While, especial fragment with size 1963 bp was detected only in DNA isolated from 40% PEG stressed plants. These fragments may activate and induce the synthesis of particular nucleotides leading to novel stress proteins. Therefore, analysis of the scanned SDS – PAGE gel proteins showed the presence of several new protein bands having low molecular weights ranging between 83.5 and 15.8 KDa which are characteristic to ABA and JA treated plants under water stress. On the other hand, the cytological effects of ABA and JA on cell division and their capacity to induce chromosomal abnormalities in maize root tips were studied. They had a pronounced effect on mitotic index. (MI), frequency of mitotic phases, percentage and types of mitotic abnormalities. The abnormalities induced by the applied concentrations are stickiness, c- metaphase, laggards, disturbed metaphase, micronuclei and multinucleate cells. These results indicate that ABA and JA induce alteration in gene expression either by induction or repression of some new genes leading to changes in protein patterns, which   play a distinct role in the mechanism of drought resistance and then increase adaptation of maize seedlings to water stress.


25/18 PRODUCTION CONDITIONS OF AN EXOINULINASE BY STREPTOMYCES GRISEUS WTA-37

W.A. El-Shouny, T.E. El-Banna*, A.B. Mohamed and A.F.H. El-Said

Botany Department, Faculty of Science and Microbiology Department, Faculty of Pharmacy*, Tanta University, Egypt

Forty nine different bacterial isolates were tested for their potentialities of inulinase production. The best producer was identified as Streptomyces griseus WTA-37 and factors affecting   inulinase production were studied. Maximum production of the enzyme (1562.5 U/ml) was obtained after 2 days at 30OC and pH 7.0. The optimal medium for the enzyme production was that devised by Lim et al. (1998) containing 0.5% inulin, 0.5% sucrose, 0.3% NaNO3, 0.3% yeast extract, 0.5% CaCl2, in addition to the induction of 24 h growing culture of S. griseus WTA-37 by 0.2% inulin.

 

26/18 MORPHOLOGY STRUCTURE AND ENZYMATIC HYDROLYSIS OF RADIATION CROSSLINKED POLY (Ε- CAPROLACTONE)

H.A. Abd El-Rehim

National Center for Radiation Research and Technology, Atomic Energy Authority, P.O. Box 29 Nasr City, Cairo, Egypt.

Crosslinked poly (ε- caprolactone) (PCL) was prepared by electron beam irradiation in the presence of the polyfunctional monomer, A-TMPT. As the A-TMPT ratio and irradiation dose increase, the PCL gel fraction increases and consequently its heat resistant property significantly improves. The thermal parameters of crosslinked PCL such as the melting temperature (Tm), recrystallization temperature (Trc), and heat of recrystallization (DHrc), were slightly influenced. The morphology and structure of biodegraded crosslinked PCL were investigated by scanning electron microscope (SEM) and differential scanning calorimetry (DSC). The results showed that the erosion reaction temperature had a great effect on the morphology and structure of modified PCL and consequently influenced its biodegradation rate. The introduction of the crosslinking into PCLdiminished its ability to biodegrade.

27/18 BIOCHEMICAL AND MOLECULAR CHARACTERIZATION OF NEW EGYPTIAN ISOLATES OF BACILLUS THURINGIENSIS.

T.M.A. El-Kawokgy; W.K. Hegazyand H.H. Salem*

Microbial Genetics Dept., NRC, Dokki, Cairo, Egypt.

*Nucleic acid Dept., Mubarak City for Sci.Res. and Techn. Applic. Alexandria, Egypt              

Seventy-one local bacterial strains were isolated on NA medium from dead snails and evaluated for their ability to produce molluscicidal toxins using bioassays against Biomphalaria alexandrina snails. The Phenotypic characterization of these isolates revealed that only one isolate (no.66) among all tested strains produced parasporal crystal in addition to oval spores, which characterized the B. thuringiensis standard strains. These results indicated that the isolate symbolized as 66 might be identified as B. thuringiensis. Restriction Fragment Length Polymorphism (RFLP) analysis for the amplified 16S rRNA genes of the tested strains which digested with four restriction enzymes (HindIII, HinfI, AluI and PstI) showed that isolated strain 66 exhibited or shared the same restriction patterns of the reference strain B. thuringiensis (tt4) and displayed closely related patterns to the other reference strain (HD522) differing only by the Alu1 restriction profile. On the other hand, the 16S rDNA RFLP profiles of the strains 71, 38, 15 and 53 were clearly different from those obtained with both reference strains (HD 522 and tt4) and the isolated strain 66. Isolated strain 66 was selected for 16S rRNA gene sequencing, and for biochemical identification. The obtained results suggested that strain 66 could be genetically related to both B. thuringiensis serovars (israelensis and tenebrionis). For this reason, strain 66 was selected as biomolluscicide agent for snails.

28/18 GENETIC IMPROVEMENT OF ALKALINE PROTEASEPRODUCTION VIA BACILLUS PROTOPLAST FUSION

E.A.MSolaiman and M. E. Moharam*

Microbial Genetics Dept. and *Microbial Chemistry Dept., National Research Center, Cairo, Egypt

This study aimed to improve the production of alkaline protease by Bacillus strains through protoplast technique. Different Bacillus strains were screened for alkaline protease production. The best producers were B. alvei and B. licheniformis (local isolates) were fused together. Twelve fusants were selected on the basis of antibiotic resistance pattern. All fusants were characterized on the basis of enzyme activity, resistance to antibiotics and different chemical and physical factors. Plasmid profiles studies of these fusants and their parents showed the transfer of some plasmids from each parent to the fusants. All fusants showed higher efficiencies in enzyme productivity than their parents. Up to five times alkaline protease activity were found in fusants comparing with their parental strains production. The enzyme overproducing fusants strains are available to be used commercially in various economic important industries.


29/18 DNA SEQUANCING OF SCHISTOSOMA MANSONI

A.H. Abdel-Tawab, M. Abdou and S.A.Shahat

Department of Parasitology, Faculty of Medicine, Al-Azhar University.

35 Hamsters were infected by S. mansoni through subcutaneous injection with a dose of 350 cercariae each. Adult worms were collected from the liver of infected Hamsters by perfusion and DNA was prepared followed by PCR, sequencing and phylogenetic analysis using parsimony (PAUP). High molecular weight DNA was recovered from all isolates.The target of the PCR primers was a fragment of approximately 500-600 nucleotides in length corresponding to a region of the 18s rRNA gene. Sequences were compared over 111 base pair region that identified polymorphisms among all isolates. All Schistosomal isolates shared identical sequence. However Schistosomal isolates have multiple base substitutions additional in target regions.
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