Vol. 20, June, 2005.

ça fait quoi si une femme prend du viagra 1/20 SOME BIOCHEMICAL CHANGES CORRELATED WITH RESISTANCE IN PENICILLIUM DIGITATUM TO FUNGICIDES WITH DIFFERENT MODE OF ACTIONS

M.B. Mahmoud and H.M.S. Khalifa

Plant Protection Department, Faculty of Agriculture,

Al-Azhar University, Cairo.

Biochemical changes in spray viagra pour femme Penicillium digitatum correlated with fungicide resistance were investigated including accumulation of carbohydrates, proteins, lipids and proline. The fungicides used in this study were tetraconazole, thiabendazole and iprodione, which differe in their mode of actions. Resistant isolates were obtained by UV radiation and the fungicides resistant isolates were coded as TetR, ThR and IpR for tetraconazole, thiabendazole and iprodione, respectively. The radiated spore suspensions were plated on PDA medium containing concentrations of the fungicides, i.e. 10 times that inhibit the growth of the sensitive isolate, and percentage of growing colonies was obtained. Biochemical analyses indicated that fungicide-resistant isolates contained higher amounts of total carbohydrates, proteins and lipids than the sensitive isolate. TetR contained total carbohydrates, proteins and lipids than ThR and IpR isolates, when the resistant isolates were grown on fungicides – free Dox's liquid medium. Determination of proline in the mycelial matrix showed that this amino acid accumulated greatly in the mycelium of the resistant isolates, and ThR and IpR contained proline somewhat higher than TetR isolate. These components increased gradually with increasing the concentrations of the fungicides, but the fungicides decreased the detected components in the sensitive isolates. Thus it could be mentioned that accumulation of carbohydrates, proteins, lipids and proline may operate the resistance mechanism of cialis et ananas P. digitatum to the tested fungicides.  

2/20 UTILIZATION OF LEATHER WASTES BY CHEMICAL AND ENZYMATIC TREATMENTS

A.S. Ismail, R.Z. Attia*, S. Abd el-Rahman** and N.H. El-Sayed*

Chemistry of Natural and Microbial Products Department, National Research Centre, Dokki, Cairo, Egypt.

*Chemistry of Tanning Materials and Leather Technology Department, National Research Centre, Dokki, Cairo, Egypt.

**Chemistry Department, Zagazig University, Zagazig, Egypt.

           

Chemical treatment of the finished leather scraps (trimmings; FLS), the major solid waste generated at finished operations of leather processing, has the potential to generate value of added leather chemicals. In Egypt, burning of the finished leather scraps generates huge amounts of toxic gases (C, N & S oxides), beside the carcinogen Cr(VI). Novel two steps process was proposed for de-dying of FLS and yielding amino acids. The first step (de-dying) was performed by different treatments with sodium chlorite and sodium bisulphite in acidic medium under different operational conditions. The second step (complete acid hydrolysis) was undergone by 6M-HCl. The favorable conditions in two steps and the yielded amino acids were determined. The enzymatic treatments following different physical and chemical treatments of different leather scraps were conducted by either pigeon manure (PM), microbial protease, commercial pepsin or trypsin enzymes. PM as a cheap and available enzymes afforded the highest hydrolysis for different soaked leather samples.

3/20 FACTORS AFFECTING OPTIMIZATION OF INDOLE ALKALOIDS PRODUCTION PRODUCED

obtenir du viagra en pharmacie BY P. CITRINUM MU-534

A.A. Barakat, A.A. El-Shanawany and A.M. Abdel-Hadi

Department of Botany and Microbiology, Faculty of Sciences,

Al-Azhar University, Assuit, Egypt.

The production of indole alkaloids by combien de temps avant prendre levitra P.citrinum MU-534 under different cultural conditions was studied. The production of alkaloids started from the 4th day of growth, and reached maximal at the end of the stationary growth phase (14 days) with 0.5 g/l of KH2PO4 at pH 5.5 during submerged cultivation (shaker). The basal medium supplemented with casein (2 g/l); NaCl (1%); CuSO4 (10 mg/l); ZnSO4 (10-50 mg/l); FeSO4.7H2O (100 mg/l) and mannitol have been substituted by sucrose as carbon source for optimal alkaloids production.

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dérivé du cialis 4/20 IMPACT OF NUTRIENT MEDIUM COMPOSITION ON TANNASE PRODUCTION BY ASPERGILLUS NIGER (BCT 21).

A. Barakat

Botany & Microbiology Department, Al Azhar University, Assuit Branch.

Assuit, Egypt.

A total of 75 fungal isolates attributed to 9 species were isolated from different 20 tea samples collected from markets in Assiut City. The fungal isolates were screened for the ability to produce tannase enzyme. Twenty- seven (36%) fungal isolates proved to be positive in tannase synthesis. Only one isolate related to achat cialis officiel Aspergillus niger (BCT 21) proved high potential in enzyme production. Ten with moderate production whilst, 16 with weak production. A comparative study on the stimulations production of extra and intracellular tannase was made from the newly fungal isolate. Extra and intra tannase production was carried out using submerged (SmF) fermentation system with different tannic acid, NaCl, tea and glucose concentrations. Results showed a maximum tannase production occurred in the culture broth containing 1.5 and 1% (w/v) tannic acid and 1.0 and 0,5 % glucose concentration for both   extra and intra tannase activity, respectively. Also, extra and intracellular tannase showed a good value at 1% NaCl .and 3.0 and 1.5 % (w/v) of tea concentrations. The optimum condition for maximum mycelial dry weight was found to be 6 % in case of NaCl and tannic acid; 2 % glucose and 1.5 % tea concentration

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5/20 GERMINATION AND BIOCHEMICAL STUDIES ON SOME DESERT PLANTS

M. Zayed, H. El Rabey* and A. Radwan**

Faculty of Science, Minufiya University. Shibin El-Kom, Egypt.                  

*Genetic Engineering and Biotechnology Research Institute, Minufiya University, Egypt and **Desert Research Center, Matariyah, Egypt.

Seed germination and seed storage proteins of Balanites aegyptiaca, Capparis decidua and Delonix elata from the South-Eastern corner of Egypt were investigated. Germination was tested using normal tap water for four weeks. Germination experiments were conducted to study the effect of sulfuric acid, and constant and alternating temperature on the germination of the three desert plants.The germination percentage was generally very low in all studied species, but it was relatively improved in C. decidua and D. elata by soaking in concentrated sulfuric acid and by raising the temperature up to 25°C under alternating dark and light conditions, whereas seeds of B. aegyptiaca failed to germinate when treated with any concentration of sulfuric acid due to the damage effect of sulfuric acid on their embryos. Seed storage proteins of the three taxa were analyzed using SDS-polyacrylamide gel electrophoresis. The proteins were analyzed twice according to their solubility in water and buffer. Thirty two protein units were resulted from the poly acrylamide gel electrophoresis of water soluble proteins and 24 protein units were resulted from the electrophoresis of the buffer soluble fraction. The protein data were analyzed using the SPSS program to elucidate their genetic relationship B. aegyptiaca and C. decidua are genetically related to each other, whereas D. elata was genetically different and appeared distant from both of them.

 

6/20 A POSSIBLE ROLE FOR JASMONIC ACID HORMON AND MYCORRHIZAL FUNGI (Glomus spp.) IN ADAPTATION OF ZEA MAYS PLANTS TO HEAVY METALS STRESS

Th.R. Mohamed

Botany Department, Girls College for Arts, Sciences and Education,

Ain Shams University, Heliopolis, Cairo, Egypt.

The effect of the interaction between vesicular-arbascular mycorrhizal fungus (VAM) (Glomus spp.) and Jasmonic acid hormone on growth, productivity, biochemical and molecular aspects of maize plants grown in the presence of chromium or zinc with three concentrations (1,2and 4mM) were investigated. There was an increasing of plant hight, number of leaves, fresh and dry weights of shoots and roots, soluble proteins and nucleic acids (DNA&RNA) contents of the yield of maize plants. The electrophoretic banding pattern of proteins extracted from treated and untreated roots revealed the occurrence of quantitative and qualitative changes .The induction of novel proteins play an integral role in increasing plant protection against the harmful effect of heavy metals .Changes in gene expression in maize plants were also investigated. The primer ATAATTAGAA was used in the PCR-RAPD analysis of genomic DNA isolated from roots of the treated and untreated maize plants with mycorrhiza and /or JA grown under the effect of chromium or zinc 4mM concentration.

 

7/20 NIROMYCIN A: AN ANTIALGAL SUBSTANCE PRODUCED BY STREPTOMYCES ENDUS N40.

S.A. El-Sherbiny, M.F. Ghaly, Y.M. El-Ayoty and N.S. Fleafil

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

The antialgal activity of 107 actinomycete isolates were isolated from different localities in El-Sharkia, El-Dakahliya and El-Meniya governorates, Egypt. Isolate No. 40 (isolated from Ekhtab, Aga, Dakahliya) was selected and identified as Streptomyces endus N40. The optimum culture conditions for the production of the antialgal metabolite were studied. It has been found that the optimum incubation temperature for the production of the antialgal metabolite from Streptomyces endus N40 was 28oC after incubation period (8 days) at pH 6.5. The most favourable carbon, nitrogen, phosphorus, microelement and vitamin sources were maltose (6g/L), asparagine (1.486g/L), K2HPO4 (1g/L), FeSO4.5H2O (0.01g/L) and (inositol) (0.01mg/L) respectively. The maximum antialgal activity was obtained against Anabaena sp., Anabaena flos-aqueae, Nostoc sp. and Anacystis nidulans. The antialgal substance had extracted using xylene solvent. Formulation and identification of the antialgal substance that produced by Streptomyces endus N40 was carrying out using IR, Mass, NMR spectra and elementary analysis and these results were emphasized that the molecular weight equal 279.33K Da. with chemical formula (C15H21NO4) and identified as Niromycin A.

8/20 THE ROLE OF MOLECULAR EVIDENCES IN JUSTIFYING THE RESPONSE OF BARLEY (HORDEUM VULGARE L.) AND WHEAT (TRITICUM AESTIVUM L.) SEEDLINGS TO LEAD ION TREATMENT

H. El Rabey and M.A. Zayed*

Genetic Engineering and Biotechnology Research Institute, Minufiya University, PO Box 79 Sadat City, Egypt.

*Faculty of Science, Minufiya University, Shibin El-Kom, Postal Code 32511, Egypt.

Nuclear genome characterization of barley and wheat were investigated using AFLP (Amplified fragment length polymorphism) markers and seed storage protein electrophoreses to explain different responses of barley and wheat seedlings treated with different concentrations of lead nitrate for different periods. The responses of barley and wheat seedlings to lead nitrate were studied as functions of lead rates (concentrations and durations of treatment), seedlings age, and the presence of other salts. An increase in the lead nitrate concentration from 10-6 to10-2 M and treatment duration from one to seven days resulted in greater inhibition of plant growth. The shoots are more tolerant to lead than the roots. The toxicity of lead nitrate was greatly reduced by the presence of other salts. The plant species under study showed species-specific quantitative differences to lead ions with respect to plant age and similar responses concerning the exposure to lead ion. 175 AFLP markers were scored in the two species, 56 out of which were common to both species, 53 were found in barley and 66 were found in wheat. On the other hand, 34 protein units were scored as a result of seed protein electrophoresis using polyacrylamide gels under reducing conditions. 13 protein units were common to both barley and wheat, 10 units present in barley and 11 units present in wheat.

 

9/20 MOLECULAR FINGERPRINTING OF SOME AFRICAN VIGNA SPECIES BASED ON RAPD ANALYSIS

M.A. Elkholy

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

Genetic polymorphism in 14 African Vigna species belonging to 7 sections of genus Vigna were   investigated by RAPD-PCR analysis. Thirteen primers were tested, only seven generated   reproducible and informative   amplified products that   were used to distinguish the studied species. A total of   95 DNA bands were detected, of these 77 bands   were polymorphic, and three were unique. Data of RAPD profiles were pooled   to estimate similarity according to Dice equation. The similarity coefficient values were used to produce a dendogram that illustrates the genetic distance among the studied species. The grouping of the studied species based on RAPD data has been discussed in the light of their taxonomic and phylogenetic relationships.

 

10/20 DEVELOPMENT OF LENTINUS EDODES " SHIITAKE " MUSHROOM PRODUCTION IN EGYPT USING DIFFERENT TYPES OF WOOD AS LIGNOCELLULOSIC SUBSTRATES

K.A. Abdel-kawi and G.A.M.A. Darwish*

Soil, Water and Environment Research Institute, Agricultural Research Center,

Giza, Egypt

*Central Lab. for Food and Feed, Agricultural Research Center, Giza, Egypt

Three strains of Lentinus edodes were studied on small scale production to identify suitable strain for production of shiitake mushroom in Egypt using Casuarina, Cupressus and Eucalyptus sawdust as source of cellulose and lignin. Sawdust substrate supplemented with rice bran in three different levels of 3, 5 and 7% on dry weight basis were inoculated with spawn vegetative growth made from strains namely L. edodes R-26 USA, L. edodes 291 ATCC 28759 USA and L. edodes 504 Ifo 3110 Japan. Biological efficiency percent (BE%) and data of mushroom harvested from the inoculated sawdust substrate at periods of 75 and 124 days, were evaluated. The best strain in BE% was L. edodes 504 ifo grown on Cupressus and Casuarinas that gave 32.03 and 24.85% at 5% of sawdust substrate supplemented with rice bran additive, respectively, followed by L. edodes 291 ATCC 28759 on Cupressus and Casuarinas with BE % of 22.0 and 19.65%, respectively at 5% rice bran additive at the same environmental conditions. Also, the chemical analyses of fruiting bodies showed   that L. edodes 504 ifo recorded 27.0, 2.6, 51.5, 8.9 and 6.1% for crude protein, fat, carbohydrate, fiber and ash, respectively. Meanwhile, total essential amino acids reached 31.3, which represents 81% of the total amino acids approximately.

 

11/20 GENETIC VARIATION OF APHANIUS SPECIES (CYPRINODONTIDAE) USING RAPD-PCR, PROTEIN ELECTROPHORESIS, AND ISOZYMES.

O.E. Khafagy

Department of Fish Resources and Aquaculture, Faculty of Environmental Agricultural Sciences, Suez Canal University.

Family Cyprinodontidae is represented in Egypt water by two species, Aphanius dispar and Aphanius fasciatus. The Killifish Aphanius dispar distributed in brakish water as well as marine water. In view of this adaptive versatility, their chromosome complement was examined to study the type of chromosomes. The use of the electrophoresis heretic analysis of the muscle proteins patterns of two species and their hybrid indicated that each of both valid species and their hybrids have distinct protein patterns. High genotypic diversity was found within populations. Among two Aphanius species dispar, fasciatus and their hybrid 21 banding positions, were resolved. the electrophoresis showed a species-specific pattern and in some cases these patterns differ of the same species. There are few characters have been detected by morphological analysis. The general genetic polymorphisms and structure of variability among the two species cyprinodont fishes and to carry out genetic variation studies based on modern techniques. To study some biological and the genetic variation among Aphanius species using protein-banding patterns muscle proteins, esterase isozyme polymorphisms and RAPD-PCR DNA markers. Restriction endonucleases of the genomic DNA have been used to detect the genetic variability among and within fish populations. It is the most successful and accurate methods. The relative from, molecular weight (MW) and band frequency fingerprints generated by the 3 primers revealed unique for each Aphanius species in terms of number and position of RAPD bands.

 

12/20 GENETIC DISCRIMINATION OF TILAPIA SPECIES BY RANDOM AMPLIFIED POLYMORPHISM DNA (RAPD).

O.E. Khafagy

Department of Fish Resources and Aquaculture, Faculty of Environmental Agricultural Sciences, Suez Canal University.

Polymerase chain reaction (PCR) analysis has proved to be very powerful in studying genetic variations at interspecific level and has been applied to differentiate the Tilapia species; Oreochromis aureus (Steindacher,1844), O. niloticus (Labroidei), Tilapia galilaeus (Ardebi,1757), T. zillii (Garvais,1848) and O. mossambicus (Peters , 1858). Five species of Tilapiini tribe (Cichlidae Perciformes) are available in Egypt. The comparative study revealed that the patterns of loci differentiated the species. The analyses showing patterns of easy interpretation, were chosen to document the interspecific differences. The present results showed that Tilapia galilaeus was greatly different than the other species, Tilapia Zillii was different from the other species, but O. aureus and O. mossambicus were nearly similar. PCR has promising value in adapting new taxonomy of different fish species depending on variations in the DNA itself, defined by using methods such as sequencing, hybridization and the study of restriction fragment length polymorphisms using restriction enzymes.

 

13/20 EFFECT OF PLANT GROWTH REGULATOR ON CALLUS FORMATION OF CYCAS REVOLUTA PLANTS

O.H. EL-Shiaty

Ornamental. Dept., Fac. Agric., Cairo Univ. Giza, Egypt.

This study was carried out at the tissue culture laboratory of Shoura Technology, Tanta, Gharbia gov. during (2003 – 2004 ) the work was planned to study the effect of different auxins (2,4 –D, NAA and NOA) and combination between 2,4-D and different cytokinins (Kin, 2ip and BA) on callus production of Cycas revoluta explants. The obtained results showed that 2,4-D was the most suitable auxin for swelling, callus initiation callus formation percentage and fresh weight specially with the concentration of 5 and 2.5 mg \ L. Moreover, the best treatment jar callus initiation and formation of two experiments were recorded when used 2,4-D levels were combined with 2ip.

14/20 MICROBIAL FLORA ASSOCIATED WITH THE WINGS OF SOME BITING AND NON–BITING DIPTEROUS INSECTS AND CHARACTERIZATION OF MOST POTENT ACTIVE METABOLITE

H.M. Mahdy and M.I. Hassan*

Botany Department and *Zoology Department, Faculty of Science, Al-Azhar University, Madient Nasr, Cairo.

Nine species of bacteria, 2 species of yeasts and 1 species of actinomycetes were isolated from wings of naturally caught the sandfly, Phlebotomus papatasi; the mosqueto, Culex pipiens; the false stable fly, Muscina stabulans and the house fly, Musca domestica. Nutrient agar amended with yeast extract and tryptose blood agar media recorded the highest number of microorganisms. Bacillus circulans 88T was the most potent organism producing active metabolite. The physicochemical properties of the purified active metabolite indicated the aromatic nature of the compound. Emperical formula of the compound was C30H37N4SO9. The antimicrobial activity was also investigated.

15/20 MICROBIOLOGICAL QUALITY OF DRINKING WATER COOLING SYSTEM (WATER SUPPLIES) IN SOME TRAINS SOLID IN EGYPT

E.M. El-Safey

Botany and Microbiology Department, Faculty of Science, Al-Azhar University (Now, on Secondment to Community College, King Saud University,

Al-majma’ah 11952),

Numerous microorganisms, including bacteria, fungi and algae, are present in water, forming a complex ecosystem. Among these organisms, bacteria and fungi are responsible for most of the contamination in drinking water and dieses that are intrinsic to the health public. On the bases of morphological and biochemical and serological we identified different bacterial and fungal isolates. Sixty-four fungal isolates (13 genus) were detected in a total 100 drinking water cooling system samples in some trains in Egypt. Suggested names including, Acremonium strictum (4%), Aspergillus flavus (2%), Aspergillus niger (2%), Aureobasidium (2%), Cladosporium (14%), Pseudallescheria boydii (12%), Fusarium cacasicum (8%), Fusarium moniliform (4%), Fusarium oxysporum (4%), Penicillium chrysogenum (8%), Penicillium corylophilum (4%), Phialophora bubakii (2%) and Phoma exigua (2%). Twenty-four bacterial isolates represent six genus were identified to species level, this including the following suggested names: Staphylococcus epidermidis (3%), Bacillus subtilis (3%), Pseudomonas aeruginos (2%), Pseudomonas pseudomallei (1%), Pseudomonas, mallei (1%), E. coli O157:H7 (2%), E. coli (type strain, 4%) Salmonella sp. (6%), (1%), and Enterobacter coloacae (2%). In addition to that seven heavy metals, Zn++, Cu++, Co++, Fe++, Pb++, and Cd++ were detected for hundred water samples under investigation. Antimicrobial susceptibility of the bacterial isolates was examined. All bacterial isolates were sensitive to Polymxin B, Tetracycline, Chloramphenicol, Amoxycillin clavulanic acid (augmentin) 2:1, Gentamycin, Kanamycin, and Sulphamethoxazole/ trimethoprim 19:1 respectively. On the other hand, four isolates were resistant to Bacitracin, one isolates was resistance to Ampicillin, four isolates were resistance to Penicillin G and two isolates were resistance to Nitrofurantion. The most effective microbial growth inhibitors were sodium azide, ZnCl2, CdCl2, NiCl2, CuC12 and FeCl2 (at 500, 200, 100, 50 μ/ml) respectively. On the other hand, EDTA and Urea, exhibited no effect at all on growth of bacterial isolates. Genus Bacillus and pseudomonas has ability to produce all extra-cellular enzymes with high productivity, genus E. coli weakly produced lipases and celluloses and do not produces either proteases or amylases, Genus Salmonella very weakly produces proteases and lipases and no produces neither cellulose nor amylases, Enterobacter, exhibited weakly producing to proteases, lipases, amylases, and not produce cellulases and genus Staphylococcus has no ability to produce extra-cellular enzyme at all. Data emphasized the fact that the train water cooling system has many microbial environmental problems, which may lead to series health problems for traveling peoples.

 

16/20 PHYSIOLOGICAL AND MOLECULAR CHANGES ASSOCIATED WITH PETIOLE ELONGATION IN SQUASH (CUCURBITA PEPO L.) INDUCED BY EPIBRASSINOLIDE

H.Kh.I. Khattab, H.F.S. Ahmed and O.M. Saleh*

Botany Department, Faculty of Science, Ain Shams University

*National Product Research Radiation Technology Center

Cairo, Egypt.

Brassinosteroids are naturally occurring plant growth regulators, which when applied exogenously evoke cell elongation and affect a number of physiological processes. In this work, we investigated the effect of soaking squash seeds in epibrassinolide (EBL) solutions (0.25 and 5 ppm) for 24 hours on certain physiological aspects. Both treatments induced increase of growth criteria in terms of branching, and fresh & dry weights of squash plants, as well as the length and circumference of petiole. The enhanced growth of petiole was concomitant with decreased levels of total simple phenols, percentage of lignin, as well as cytosolic and cell wall calcium. This decline was inversely proportional to the applied concentration of EBL. Treatments also quantitatively enhanced total soluble proteins of petiole, whereby qualitatively a new protein (Mr: 59.2 kDa) was induced, in response to the high concentration of EBL and two (Mr: 39.8 and 28.4 kDa) in treatment with its low concentration. In addition, using the RAPD-PCR technique, genomic DNA fingerprinting in petioles was compared in cases of the untreated and the EBL-treated plants. Nine of the ten arbitrary decamer primers used showed good amplification coefficients in petioles of the untreated and treated plants. Some polymorphic qualitative fragments were shown with two of the used arbitrary primers. The molecular sizes (bp) of the amplified DNA fragments were also shown. Thus, we concluded that EBL-promoted elongation in squash petioles is assumed to act via modulation of gene expression and protein patterns and that would affect cell expansion and some related metabolites.

 

17/20 BIOLOGICAL AND MOLECULAR STUDIES ON AN EGYPTIAN ISOLATE OF BEET CURLY TOP GEMINIVIRUS

S.Y.M. Mahmoud; A.M. Galal* and M.H. Abdel-Ghaffar**

Agricultural Botany Departement (Microbiology), Faculty of Agriculture, South Valley University, Souhag, Egypt.

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

** Agricultural Microbiology Departement (Virology Lab.), Ain Shams University, Cairo, Egypt.

Beet curly top geminivirus (BCTV) is widely spread in the main growing areas of sugar beet (Beta vulgaris L.) in Egypt. Natural infection and spread of BCTV syndromes in beet plants in Kafr El-Sheikh Governorate were assessed visually for the presence of BCTV from December 2003 to May 2004. The numbers of plants showing symptom were increased by time. The virus isolate was identified as BCTV by symptomatology, mode of transmission, purification, electron microscopy, and by the polymerase chain reaction (PCR). The virus was successfully transmitted by syringe injection and by Empoasca decipiens leafhopper but not with Bemisia tabaci. This virus was transmitted to plants belonging to Chenopodiaceae, Leguminaceae and Solanaceae. Virus particles in monomers (20 nm) or dimers with dimension of about 20x30 nm were observed when the purified preparations from BCTV-Phaseolus vulgaris examined by transmission electron microscopy. Purified virus yield was about 1.3-1.5 mg/kg infected plant tissues. BCTV-DNA was detected in the purified virus preparation and infected sugar beet samples by PCR using specific primers to amplify BCTV-coat protein gene with a size length of approximately 800 bp. These data demonstrated the presence of BCTV in Egypt by using specific and highly efficient methods. The results indicated that there no serological relationship was observed between tomato yellow leaf curl virus (TYLCV) and BCTV. Greenhouse studies indicated that sugar beet cv. Pleno was more susceptible than cv. Gazell. Younger plants showed greater susceptibility to infection than older ones. Application of lemongrass oil vapor led to significant decrease in the percentage of BCTV transmission by leafhopper. These results suggested also the use of lemongrass oil vapors, with plants that are susceptible to BCTV infection. It is suggested be an effective alternative component for BCTV control programes.

18/20 EFFECT OF PROBIOTIC BACTERIA ON SOME PROPERTIES OF STIRRED YOGHURT LIKE PRODUCT

M.M. Ashour, M.Z. El-Abbassy, A.A. Abd El-Galeel and K.N. Eskandar

Food Science Department, Faculty of Agriculture, Zagazig University

Stirred yoghurt (sy) and probiotic stirred yoghurt like products (psylp) were manufactured from cows’ milk (3% Fat) by using yoghurt starter and probiotic starter (ABT-3 and ABT-5) cultures respectively. The starter cultures were added at a level of 0.5% to the milk. The (sy) and (psylp) samples were chemically, microbiologically and sensory analysed. The total bacterial counts of (sy) were higher than (psylp) when fresh and during the storage period up to 7 days. Yeasts & moulds, spoilage and pathogenic bacteria were not detected in all (sy) and (psylp) samples when fresh and during the storage period. No differences in the gross chemical composition between (sy) and (psylp) were observed. However, the acidity of (psylp) was higher than (sy). Total scores of the organoleptic properties of both (sy) and (psylp) were relativelly similar. But (psylp) containing ABT-3 showed slightly higher total scores than other samples. It could be concluded that using ABT-3 culture in the manufacture of stirred yoghurt is recomended.

 

 

19/20 BIOCHEMICAL STUDIES ON FREE RADICAL SCAVENGING AND PROTECTIVE ACTIVITY OF AMBROSIA MARITIMA, L., RUTA GRAVEOLENS, L. AND ZINGIBER OFFICINALE, ROSC.

A.M. Abdel-Samad;M.M. Farag; K.M. El-Deib*; M.M. Abdel-Miseh*

and M. M. Ahmed*

Biochemistry Dept., Fac. of Agric., Cairo Univ., egypt.

*Molecular drug Evaluation Dept., National Organization for Drug, Control And Research (NODCAR), Egypt.

Extracts of aerial parts of Ambrosia maritima, leaves of Ruta graveolens and rhizomes of Zingiber officinale were assessed for inhibitory effects on the DNA-sugar damage induced by Fe++ / ascorbate model system.The study was extended to evaluate the free radical scavenging activity of these extracts using Diphenylpicrylhydrazyl radical (DPPH). Our results showed that ethanolic and water extracts (Et1, W1) of these plants protected DNA against oxidative damage of its deoxyribose sugar moiety. They exerted also free radical scavenging activity against DPPH radical. Successive extracts of these plants by different solvents petroleum ether (PE), chloroform (Chl), ethyl acetate (EA), ethanol (Et2), and water (W2) were also tested. The protective effect of PE of Z. officinale by three concentrations had the highest activity. On fractionating PE extract of Z. officinale into acetone insoluble fraction and acetone soluble fraction, the later was proved to be the most active one.

 

20/20 EFFECT OF SOME BIOTECHNOLOGY DRUGS ON DNA, RNA AND PROTEINS IN RAT KIDNEY.

Z.A. Teleb, K.M. El-Dieb, N.Z. Ahmed and M.A. Mohamed

Molecular Biology Department National Organization for Drug Control and Research “NODCAR”

Two doses of each drug (100 & 250 U/kg.b.w for recombinant human erythropoietin   “EPO”, 0.005 & 0.015 U/kg .b.w for recombinant growth hormone “GH” and 0.05 & 0.125 U/kg.b.w for rh-insulin) were daily injected subcutanously in rats for one week. Then kidneys were isolated, protein and nucleic acids were determined. Genomic DNA was extracted and purified for loading on 1% agarose. RNA was extracted and purified on agarose 1.3%. In addition, proteins were identified by SDS / PAGE technique. The results indicated that genomic DNA from rat kidney after one-week injection of these biotechnology drugs showed unchanged effect on the molecular size of DNA molecules, which have a molecular weight more than 10 kbp by agarose 1%.   Protein patterns on SDS/PAGE revealed the same proteins with the same molecular weight between 14 – 66 kd compared to untreated control group. RNA showed no difference in the molecular size between all treated samples after a short-term period. RNA molecules have molecular weight less than 2.5 kbp.

 

 

21/20 ENHANCED PRODUCTION AND PROPERTIES OF A SURFACTANT BY A GAMMA RAY-INDUCED MUTANT OF BACILLUS SUBTILIS

A.S. Bashandy; H.M. Abu Shady*; N.H. Aziz and H.M.M. Ibrahim

Microbiology Department, NCRRT, AEA

*Microbiology Department, Faculty of Science, Ain Shams University, Cairo, Egypt

Surfactin production by Bacillus subtilis AB01335-1, AB02238-1 strain   was used to examine the effect of altering salt concentration, pH and temperature on surfactin activity. Experimental variables included NaCl (0-15% w/v), pH (3-10) and temperature (20-100oC). Each of these parameters as well as selected combinations were investigated. The surface-active compound retained its properties during exposure to elevate temperatures (100°C), high salinity (8 % NaCl) and a wide range of pH values. The production and properties of a biosurfactant synthesized by Bacillus subtilis AB01335-1, AB02238-1 strain, using molasses as substrate were investigated. The microorganism was able to grow and to produce surfactant on molasses waste. A gamma ray-induced mutant of Bacillus subtilis strain AB01335-1, AB02238-1, capable of hyperproduction of biosurfactant from hydrocarbons, was isolated and named as Bacillus subtilis AB01335-1M4 and Bacillus subtilis AB02238-1R2. The stability of the mutant strains was observed after 6 month of subculturing. The mutant strains Bacillus subtilis AB01335-1 M4 and AB02238-1 R2 showed 5 and 2 times more biosurfactant production as compared to the parent when grown on minimal medium, and increase in surfactant activity by the mutants was also observed as indicated by haemolysis and drop collapsing assay. Enhanced biosurfactant production was also investigated in the mutants compared by the parent when grown on molasses.

 

 

22/20 UTILIZATION OF WHEAT GERM IN THE PREPARATION OF BABY FOODS FORMULA

M.M. Ashour, M.Z. El-Abbassy, A.A. Abd-Elgaleal and A.E. Ahamed

Food Science Department, Faculty of Agriculture., Zagazig University

Mothers depend on baby foods either wholly or partially especially in the early stages in the baby’s life. This need for baby foods may be due to many factors, the insufficient quantity of milk secreted by the mother, which is not able to face the requirement of the new born baby. Also it may be due to the engagement of mothers in work. At the beginning of the 4th month of the baby’s life the mothers start to feed him on baby foods containing cereals, fruits and vegetables, up to the weaning age. In the present study, the chemical, microbiological, mycological and sensory evaluation was carried out on some baby foods formula available in the local market and prepared wheat germ formula. It was found that the prepared wheat germ formula containing milk powder and sugar had the following chemical composition 33.2 %   protein, 56 % carbohydrate, 16.4 % fiber, 12.10 % fat and 4.9 % ash. The samples of Nestle with wheat contained about 7.9 % protein, 84.6 % carbohydrate, 2.10 % fiber, 0.88 % fat, 1.71 % ash and 2.04 % moisture. The samples of Cerelac with fruits contained about 15.09 % protein, 69.4 % carbohydrate, 2.42 % fiber and 9.01 fat. While the samples of Cerelac with vegetables contained about 15.04 % protein, 67.6 % carbohydrate, 3.09 % fiber, 9 % fat, 2.38 % ash and 2.534 % moisture. The samples of Riri with vegetables contained about 15.03 % protein, 76.4 % carbohydrate and 1.51 % fat, while the samples of Riri with cereal milk contained about 16.05 % protein, 71.66 % carbohydrate, 5.09 % fat and 5.06 % moisture. The samples of Gerber with fruits contained about 19.46 % carbohydrate and 0.86 % fat, while the samples of Gerber with vegetables contained about 0.82 % protein and 8.81 % carbohydrate.

 


23/20 BIOLOGICAL CONTROL OF CERCOSPORALEAF SPOT (CLS) DISEASE OF SUGAR BEET AND ITS ASSOCIATED FUNGI

M. Hashem and E.S. Farrag*

Botany Department, Faculty of Science, Assiut University.

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

Cercospora leaf spot (CLS) has become one of the most common and sever diseases on sugar beet plants at Kafr El-Sheikh Governorate in recent years. The causal organism (Cercospora beticola) and other associated fungi were identified. In vitro, Epicoccum nigrum exudates inhibited conidiophores development and conidial formation of Cercospora beticola. In addition, antagonistic effect of E. nigrum against the most frequent species associated with this disease was proved via antibiosis. Spore germination of Alternaria alternata, Drecheslera spicifera and Fusarium avenaceum was significantly inhibited when treated with E. nigrum exudates. Under field conditions application of E. nigrum exudates as a spray successfully controlled the Cercospora leaf spot (CLS) on sugar beet cvs “Top” and “Ras poly”. Disease severity (%) of CLS on shoot system of both cultivars was significantly reduced. Top and root yields were greatly stimulated. Enhancement in sucrose content and its purity as well as total sugar yield in both cultivars were good indicators for the efficiency of treating of sugar beet leaves by E. nigrum culture filtrate.

 

24/20 QUALITY CONTROL OF SOME BIOTECHNOLOGY PRODUCTS, IDENTIFICATION AND MICROBIOLOGICAL TESTS

F. H. Ahmed and E. H. Ibrahim*

Microbiology Department, National Organization for Drug Control and Research (NODCAR) and *Research and Biotechnology Department, Center for the Control of Biological Products & Vaccine (NRA)-NODCAR, Cairo, Egypt.

Quality control of vaccines, blood products and biotechnology products is very important to monitor safety and quality of the products. In this study some bioengineered products (Interferon, Erythropoietin, Granulocyte macrophage colony stimulating factor, Insulin, and Growth hormone) were quality controlled as examples of biotech biological products. The products were subjected to potency test (Interferon and Erythropoietin), viral safety test (Erythropoietin), bacterial endotoxin, microbiological contamination, purity, and identity tests (Interferon, Erythropoietin, Insulin, Granulocyte macrophage colony stimulating factor, and Growth hormone). Results showed that all the products were sterile and free (or within specified limit) of endotoxin. Both Interferon and Erythropoietin were shown to be potent. Purity test revealed that all products were pure. Viral safety test revealed that Erythropoietin is free of viral contaminations. This study showed that quality control tests to monitor the safety and efficacy of biotech biological products should be performed.

 

 

25/20 HYGROMYCIN A PRODUCTION BY IMMOBILIZED CELLS OF STREPTOMYCES HYGROSCOPICUS NRRL 2388

E.E. Habib and R.A. El-Domany*

Microbiology Departments, Faculty of Pharmacy, El-Mansoura

and *El-Minia Universities, Egypt

Hygromycin A, an antibiotic produced by Streptomyces hygroscopicus NRRL 2388. It 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 cattle dysentery. Reported herein are the results of the production of hygromycin A by free and immobilized cells of Streptomyces hygroscopicus. The cells were immobilized with calcium alginate and sponge support materials. Low yield of antibiotic (520 mg / ml) was obtained when calcium alginate beads were used after 7 days of cultivation compared to the free or other immobilized cells. The maximum amount of hygromycin A (1100 mg / ml) was obtained after 8 days of cultivation at an initial pH value of 7.0, when the cells were adsorbed on the sponge compared to 700 mg / ml of hygromycin A obtained when the free cells were used after 6 days of cultivation. During repeated batch operation, sponge immobilized cells exhibited higher hygromycin A production than Calcium alginate immobilized cells under similar experimental condition.

26/20 MUTAGENIC EFFECTS OF WATER EXTRACT OF HIBISCUS SABDARIFFA L.ON VICIA FABA PLANT.

N.R. Abd-Elhamied

Genetic & Cytology Dept., National research center, Doki, Cairo, Egypt.

The present study was designed to investigate the cytogenetic and mutagenetic effects of water extract of the medicinal plant Hibiscus subdariffa. L, belonging to family Malvaceae on both mitotic and meiotic cells of Vicia faba plants. This study further includes the action of this plant extract on alterations in electrophortic profile of C2 seed storage protein of Vicia faba. The cytology of the C2 plants was also investigated. The applied extract induced a significant or highly significant reduction in mitotic and meiotic activity of Vicia faba plants and reported a variety of chromosomal aberrations represented by stickiness, laggards, fragments, bridges, unseparated and disturbed types. The mutagenic potentialities of the tested extract has been manifested by also alteration in electrophoretic seed protein banding profile.

27/20 BIODEGRADATION OF POLYCYCLIC AROMATIC HYDROCARBONS FROM TWO PETROLEUM CRUDE OILS

M.A. Abostate and Y.M. Moustafa*

National Center for Radiation Research and Technology, Cairo, Egypt.

*Egyptian Petroleum Research In Statute, Cairo, Egypt.

Two crude oils (light and heavy) were used to evaluate the microbial potential for degradation of 14 polycyclic aromatic hydrocarbons (PAHs) listed by the United States Environmental protection Agency (US-EPA) using high performance liquid chromatography (HPLC). Bacillus megaterium; Alcaligenes eutrophus; Rhodococcus rhodochrous; Pseudomonas oleovorans; five of their mixtures; Two isolated fungal strains, AspergillusSp. Strain MA-97 and Penicillium Sp. Strain MA-7 and their mixture were used in this study. All microbial treatment were capable of degrading naphthalene completely (100%) in both light and heavy oils. P. oleovorans was the most efficient bacterial strain in degrading fluorene of heavy crude oil after 15 days and phenanthrene and pyrene of light crude oil after 30 days. P. oleovorans degraded 60%and 74% of total PAHs of light crude oil after 15 and 30 days respectively. Meanwhile bacterial mixture III degraded 64% and 69% of the heavy crude oil at 15 and 30 days respectively. The highest percentage of degradation was recorded by the fungal mixture with light crude oil after 15 and 30 days (72% and 89%) respectively.

28/20 SEPARATION OF ERGOLINE ALKALOIDS FROM CELL SUSPENSION CULTURES OF IPOMOEA VIOLACEA L. (CONVOLVULACEAE) BY CAPILLARY ELECTROPHORESIS

Y. Hussein and E. Leistner*

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

** Institute of Pharmaceutical Biology, Bonn University, Germany.

Ergoline alkaloids (chanoclavine-I, lysergic acid amide (LSA), isolysergic acid amide, agroclavine, elymoclavine, lysergol, or ergometrine) were not detected by thin layer chromatography (TLC) or capillary electrophoresis (CE) in cell suspension cultures of Ipomoea violacea (morning glory) cv. heavenly blue (Convolvulaceae) grown in sterile BDS and LS media, while Ipomoea violacea seed was reported to produce ergoline alkaloids. Ipomoea violacea callus was initiated and maintained in BDS medium supplemented with Dicamba (1.0 mg/l) and kinetin (1.0 mg/l) and LS medium supplemented with 2,4-D (0.2 mg/l) and NAA (0.2 mg/l). Each medium containing 3% sucrose and 0.4% phytagel. The inoculated BDS and LS media were incubated at 25 oC under different light conditions (in the light, in the dark or in a 10 hr photoperiod). Results of CE also showed that no chanoclavine-I, LSA or ergometrine have been detected in cell suspension cultures of Ipomoea violacea grown in BDS medium supplemented with 100 mg/l L-tryptophan and 1.0 mg/l mevalonic acid (precursors of ergoline alkaloids). The limit of detection was 2.5 ng for chanoclavine-I, 25 ng for ergometrine and 50 ng for LSA. The variation in media composition, growth hormones and light conditions influenced the growth ofcalli and cell suspension cultures of Ipomoea violacea, but had no effect on the in vitro production of ergoline alkaloids.

29/20 ANTIRADICAL PROPERTIES OF OAT (AVENA SATIVA L.) FLOUR EXTRACTS

N.S.A.M. Khalil and G.A.M.A. Darwish

Central Laboratory for Food and Feed, Agricultural Research Center, Giza, Egypt

Oat (Avena sativa L.) is a source of many compounds that exhibit antioxidant activity. Vitamin E (tocols), phytic acid, phenolic compounds, and avenanthramides are the most abundant antioxidants in oat, and flavonoids and sterols are also present. To our knowledge, there is limited literature on the profile of antioxidant phytochemicals in the local oat. Hence, the objective of this study was to investigate the antioxidant property of oat flour extracts in vitro and also to identify the fractions responsible for the antioxidant activity. In the present study, we analyzed the antioxidative properties of various extracts (water, 60% methanol, 60% ethanol, and 60% t-butanol) of oat flour employing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) model system. Phenolics, antiradical and antioxidant activities were also examined in chromatographic sub-fractions of the soxhlet methanolic extract. Our results indicated that the various extracts exhibited significant antioxidant activity that did not correlate with the phenolic content. Further, two sub-fractions eluted with methanol and acetone/methanol were found to possess strong antioxidant activity in two assay systems. Our results suggest that a diet rich in oat may be useful in combating diseases in which free radical production plays a key role.

 

30/20 ENHANCEMENT OF BIOSYNTHESIS OF XYLITOL PRODUCTION BY LOCAL YEAST ISOLATES

A.A. Abou Zeid; T.M. El-Mongy*; Y.A. El-Zawahry*; M.Z. El-Foul*

and A.B. Abd El-Aziz*

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

*National Center for Radiation Research and Technology, Nasr city, Cairo, Egypt.

Six isolates out of 50 yeast colonies isolated from different fruits were found to have the ability of producing xylose reductase. They were found to be related to four genera and were identified as two of Debaryomyces hansenii, one from each of Candida shehatae, Candida tropicalis, Candida Peltata and Zygosaccharomyces rouxii. The six yeast strains were screened for production of xylitol from D-xylose. The highest three isolates (Deb. hansenii, Can. tropicalis and Can. shehatae) were selected for optimizing their fermentation conditions beside standard yeast Can. guilliermondii NRRLY-488 in comparison with the local isolates. The optimum incubation temperature for maximum xylitol production for Can. guilliermondii, Can. tropicalis and Deb. hansenii was 30oC, while their optimum initial pH value was 5.5 producing 38.3, 40.8 and 36.4 g/l xylitol from 60 g/l D-xylose after 72h respectively. Meanwhile, the optimum xylitol production for Can. shehatae was at 37oC and initial pH 4.5 after 72h recording 32.2 g/l. Ammonium chloride and urea were the most suitable inorganic and organic nitrogen sources for Can. guilliermondii and Can tropicalis for producing xylitol recording 40.9, 42.0 and 46.2, 44.0 respectively. Meanwhile, fermentol as organic nitrogen source gave best results for Can. shehatae (40.0 g/l) and Deb. hansenii (45.3g/l). Increasing the xylose concentration up to 250 g/l in the fermented medium using the best culture conditions for each isolate increased the xylitol yield for all of them and the local isolate Can. tropicalis gave the best results (160.6 g/l) over the other isolates even the standard strain Can. guilliermondii (152.0 g/l).

31/20 EFFICIENCY OF STABILISATION POND TECHNOLOGY IN BIOLOGICAL WASTEWATER TREATMENT FOR RE-USE IN AQUACULTURE AND IRRIGATION PURPOSES IN SUEZ, EGYPT.

S.A. Rabeh, S.Z. Sabae and E.A. Azab*

National Institute of Oceanography and Fisheries, Egypt

* Division of Microbiology, Botany Dep. Fac. Sci., Tanta

Raw sewage from the City of Suez were diverted partially and treated by waste stabilisation pond (WSP) system. The raw sewage passed through a series of ponds (anaerobic pond with theoretical retention time 12 hr., facultative pond with theoretical retention time 15 days, 2 maturation ponds with theoritical retention time 7 days each) before being used for fish stocking in 2 fish ponds. The final effluent was then re-used for irrigation. The pond system overall efficiency for Saprophitic (SB) , parasitic bacteria (PB) , total coliform (TC) , faecal coliform (FC), faecal streptococci (FS), Salmonella and Shigella as well as Pseudomonas aeruginosa removal values were 94.44 %, 94.64 %, 100 %, 100 %, 96.4 %, 99.99 % and 92.49 %, respectively. These results encourage the use of WSP as a cost-effective method for wastewater treatment in Egypt. Faecal coliform numbers in the final effluent were much less than 103­­/100 ml (ranged from 0.0-95/100 ml). Thus, according to WHO (1989) the bacteriological quality of the treated wastewater is suitable for both unrestricted irrigation and fish culture.

 

32/20 RAPD-PCR ANALYSIS OF SOME ASPERGILLUS FLAVUS STRAINS ISOLATED FROM DIFFERENT SOURCES

M.A. Swelim

Botany Department, Faculty of Science, Benha, Zagazig University, Egypt

The present investigation was conducted to study the differences between the six isolates of Aspergillus flavus to distinguish if they have monomorphic or polymorphic relationships. Two primers 3 and 6 were used in the RAPD-PCR analysis. The generated RAPD profile using primer 3 showed a considerable polymorphism among the studied isolates which three strains could by polyphylogenic and the other three isolates are monophylogenic. The data derived from primer 6 clarified clear discrimination among the six isolates which classified them into five groups, two groups includes two polygeneric isolates and the other two groups, each one comprise two monogeneric isolates. RAPD data were used to construct a dendrogram that estimates the relationships among the six isolates of A. flavus. Based on this dendrogram, the isolates were classified into two main groups, each one includes three isolates and the highest similarity (60%) was recorded between the two isolates collected from soil cultivated with tomato and nail using primer 3, while the six isolates could be clustered into five groups using primer 6. The highest similarity value (83.6%) was observed between the two Aspergillus flavus samples isolated from nail and coconut. On the other hand, the lowest similarity value (11.7%) was scored between the two strains of A. flavus isolated from Assiut soil and nail.

33/20 EFFICACY OF AQUEOUS EXTRACT OF THYMUS VULGARIS AN ANTIBACTERIAL AGENT AGAINST LISTERIA MONOCYTOGENES IN SOFT CHEESE

F.A. Fathy, A.B. Abd El-Khalek and S. Abd El Ghani

Dairy Science Department, National Research Centre, Dokki, Giza, Egypt

The aim of the investigation was to determine the efficacy of Thymus vulgaris aqueous extract against Listeria monocytogenes in soft cheese preservation as an additional barrier in the new concept of hurdle technology. Results showed that 13 varieties of market cheeses were hospitable for L. monocytogenes growth when artificially contaminate the opened package kept at 8-10°C or at abuse temperature of 30°C. Aqueous extract of the thyme used at 15% (v/v) inhibited the growth of the pathogen in tryptose soy broth (TSB) and milk at both storage temperatures used. The inhibition was more pronounced in TSB than in milk tubes both supplemented with the spice. In addition, results highly demonstrated the efficacy of 15% (v/v) of aqueous extract of thyme (traditionally named Zaatar) as an additional barrier in soft cheese to protect consumers especially the immuno-compromised, elderly and young persons.

34/20 EFFECT OF HOMOGENIZION AND REFRIGERATION STORAGE OF BUFFALO`S MILK ON THE QUALITY OF YOGHURT

M.A. El-Sayed, H.A. Fahmy* and A.M. Abdel-Salam

Dairy Science Department, National Research Centre, Cairo, Egypt.

*Department of Special Foods, Food Technology Research Institute, Agricultural Research Centre, Giza, Egypt.

In this study chromotographic techniques was used for the determination of   carbohydrates, organic acids, vitamin C, retinol , α-tocopherol   andviability of lactic acid bacteria in yoghurt   manufactured from homogenized and un-homogenized buffaloe`s milk. The samples used in the present study to explore the effect of homogenization and cooling storage on the components of manufactured yoghurt. Fresh yoghurt were kept under refrigeration at 6ºC ±2 for 7 and 11 days. The obtained pH results showed a significant decrease of   pH values and a slight decrease of total solids during storage in yoghurt manufactured form un-homogenized and homogenized buffalo’s milk. The obtained results revealed that there are a decrease in lactose levels and increase in glucose, galctose and organic acid levels. The data showed a slight decrease in retinol, α-tocopherol and the viability of lactic acid bacteria during fermentation and storage in yoghurt manufactured from homogenized and un-homogenized buffalo’s milk.

 

35/20 LEVELS OF SOME SELECTED ANTIOXIDANT ORGANIC ACIDS, ALDEHYDES, ALCOHOLS AND AMIDES IN OAT (AVENA SATIVA L.) GROATS AND HULLS

N.S.A.M. Khalil and G.A.M.A. Darwish

Central Laboratory for Food and Feed, Agricultural Research Center, Giza, Egypt

Research was initiated to measure antioxidant activity (AOA) of extracts from oat (Avena sativa L.) groats and hulls and the concentrations of some selected organic substances that may contribute to antioxidant activity. Antioxidant activity (AOA) of ethanolic extracts of oat fractions obtained from four egyption govrnorates namely, Behira, Giza, Ismailia and Dakahlia was evaluated by an in vitro assay that measures the inhibition of coupled autoxidation of linoleic acid and b-carotene. Total phenolic content was determined using Folin and Ciocalteau’s phenol reagent and was expressed as gallic acid (GA) equivalents. Different organic acids, aldehydes, and amides as well as one alcohol were separated by reversed-phase HPLC and detected at 290 nm. Peaks were identified by comparing retention times and spectra with known standards and verified with internal standards. Groats had significantly higher antioxidant activity than hulls. For two governorates, total phenolic content was similar in groats and hulls, whereas one governorate had higher and another lower total phenolic content in groats than hulls. Fourteen organic compounds were separated and identified in extracts; eight organic acids, two aldehydes, one alcohol (flavan-3-ol) and three amides (aventhramides). The concentrations of many of these compounds differed among governorates and between fractions. In general, caffeic acid and the avenanthramides were predominantly found in groats, whereas many of the other phenolics were present in greater concentrations in hulls.

36/20 PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OFTWO SOLUBLE RESPIRATORY C-TYPE CYTOCHROMES FROM DISSIMILATORY IRON REDUCER GEOBACTER METALLIREDUCENS

E. Afkar

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

Two soluble respiratory cytochromes c with markedly different molecular masses (55 and 80 kDa) from the bacterium Geobacter metallireducens, which uses anaerobic iron respiration as its sole energy source. The 55 kDa cytochrome c is purified to an electrophoretically homogenous state. Its molecular weight was estimated to be 55 kDa on the basis of amino acid composition and SDS-polyacrylamide gel electrophoresis analysis. Its mid-point redox potential at pH 7.0 was determined to be -189 mV.   The purified molecule binds 18-nmol heme c per mg of protein suggesting that the purified protein is monoheme cytochrome c. The 55 kDa-cytochrome c showed absorption peaks at 530, and 410.3 in the oxidized form and 551.5, 522.5, 420.1 in the reduced form. The 80 kDa cytochrome c was partially purified by two successive steps of anion exchange chromatography and ammomium sulfate fractionation up to 55% saturation followed by two steps of gel filtration. Although the 55 kDa cytochrome c showed no iron reducing activity, it can act as electron donor to the membrane bound iron reductase. No iron reductase activity could be detected when NADH used as electron donor.

37/20 LOCALIZATION OF THE DISSIMILATORY ARSENATE REDUCTASE (DASR) IN SULFUROSPIRILLUM BARNESII STRAIN SES-3

E. Afkar

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

Sulfurospirillum barnesii strain SeS-3 is one of the recently known bacterial isolates that can obtain energy to support growth by respiring the toxic oxyanions of arsenic and selenium under anaerobic conditions. The ultimate goal of this investigation is to localize the active sites for the reduction of arsenate/arsenite as well as selenate/selenite in S. barnesii strain SeS-3. The results of this study showed that the mobilization of the toxic oxyanions of arsenic and selenium by Sulfurospirillum barnesii strain SES-3 is linked to the membrane. The enzyme is specific for the reduction of arsenate, selenate, selenite, nitrite, thiosulfate and phosphate.   Whereas, no specificity was detected for arsenite nitrate, fumarate, when they served as the final electron acceptor. This study concluded that the mechanism of arsenate reduction by S.barnesii strain SeS-3 is connected into the membrane. The environmental significance of this bacterium and its impact to the bioremediation potential in the underground water and sedimentary environment is also discussed


 

38/20 Genetic Diversity and Molecular Characterization of Medicago polymorpha L. varieties

M.A. Karam and H.A.R. Deif*

Botany Department, Faculty of Sciences, Cairo University, El-Fayom Branch

*Botany Department, Faculty of Sciences, Alexandria University

Seeds of 26 Medicago polymorpha L. accessions representing 12 populations distributed allover a wide environmental range were used to assess the genetic diversity and relationships among M. polymorpha L. var. brevispina (Benth.) Heyn, M. polymorpha var. polymorpha L. and M. polymorpha var. vulgaris (Benth.) Shinners. Six isoenzyme systems and seedling proteins were shown using gel electrophoresis.A total of 15 isoenzyme loci were detected and their allele frequencies were calculated. Intra- and inter-variety genetic diversity were quantified by computing mean alleles per locus, percentage of polymorphic loci, expected average heterozygosity, genetic divergence and genetic distance using PopGen32 software. The data revealed that the populations of M. polymorpha var. brevispina collected from Australia and Chile in addition to those of M. polymorpha var. polymorpha collected from Turkey and Egypt had the highest values of genetic diversity measures indicating an interesting genetic potential for selection and may provide a valuable genetic resource for breeding programs. Based on proteins and isoenzymes data, the cluster analysis supports the traditional classification of the three varieties. Isoenzymes as molecular marker revealed variety specific loci at Est2 for M. polymorpha var. brevispina and at Amy3 for M. polymorpha var. polymorpha. SDS-PAGE of proteins distinguished M. polymorpha var. polymorpha and M. polymorpha var. vulgaris by the protein band with 70 KD and M. polymorpha var. vulgaris by the protein band with 18 KD.

39/20 CORRELATION OF ENTEROTOXINS PRODUCTION AND MULTIDRUG-RESISTANCE WITH PLASMIDS IN CLINICAL STRAINS OF STAPHYLOCOCCI

H.A. Abdel-Salam, E. El-Masry, G.H. Shaker and G. Enan*

Department of Microbiology and Immunology, Faculty of Pharmacy

and *Science, Zagazig University, Egypt.

Nosocomial infections are a significant cause of morbidity and mortality throughout the world. In developing countries it is difficult to carry out effective surveillance and control programs for this type of infection. The production of staphylococcal enterotoxins (SE) type SEA, SEB, SEC, and SED and toxic shock syndrome toxin (TSST) was studied in 26 and 31 identified clinical strains from oral and skin swabs of neonates, respectively. The characterized enterotoxigenic strains isolated from oral and skin of neonates represent 43% and 13% of total isolates, respectively. The strains produced enterotoxins were biotyped as S. aureus, S. cohnii, S. epidermidis, S. hominis and S. gallinarum. The enterotoxigenic isolated staphylococcal strains showed higher tendency to produce Dnase and coagulase. Out of 15 enterotoxigenic tested isolates, 11 (73%) were DNase-positive and 13 (87%) were coagulase-positive. Only SEC, SEB and TSST were produced and secreted to the culture medium, in addition to simultaneous production of two the enterotoxins C and B by some isolates. The enterotoxigenicity was detected by reverse passive latex agglutination (RPLA), silver-stained polyacrylamide gel electrophoresis and western immunoblot techniques. The strains positive by RPLA were also positive silver-stained polacrylamide gel and western immunoblot. Antimicrobial resistance patterns were heterogenous, since as many as 10 different patterns were characterized among 11 isolates from oral and skin swabs of neonates. The plasmids were characterized and each isolate has only one plasmid. The approximate molecular weight of plasmids of ES ranged from 9.5-32 kb, with predominance of 9.5, 12, 29 kb plasmids. The antimicrobial resistance patterns distinguished S. aureus more than did plasmid. The isolates produced the same enterotoxins showed different antimicrobial resistance patterns and insignificant plasmids. It could be concluded that there was no consistent relationship between production of enterotoxins, type of enterotoxin, antimicrobial resistance and plasmids patterns.

 

40/20 UTILIZATION OF SLENDID – TYPE ®200 AS A FAT REPLACER IN LOW FAT FATA CHEESE

H.H. Arafat*; A.M. El Fak; M.M. Nasr*; A.H. Gerguis and M.M. Ashour

Food Science Dept, faculty of Agric., Zagazig univ.

*Animal production institute Agricultural research center Dokki, Cairo

Low fat Feta cheese was manufactured from buffaloe's milk retentate and the fat replacer slendid ®200 was added in 0.2%. The addition of enhanced the quality of cheese when compared with control. The chemical composition was not affected by the addition of slendid. The Microbiological quality was affected in a better way. With respect to the rheological quality as elasticity and apparent viscosity was better in compare with control. In connection to microstructure the cheese showed a close compact protein matrix all, of the above mentioned results were observed while the cheese was fresh but after storing the cheese for 25 days, the chemical and microbiological composition were changed, and the rheological and microstructure of cheese became better. The obtained data revealed that the use of 0.2% slendid ®200 produced a better low fat feta cheese with good properties.
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