Vol. 17, June, 2004.

1/17 BIOREMEDIATION POST-PHOTOOXIDATION AND COAGULATION FOR BLACK LIQUOR EFFLUENT TREATMENT

S.M. Helmy*, S. El-Rafie and M.Y. Ghaly

*Department of Microbial Chemistry and Department of Chemical Engineering

National Research Center, Dokki, Cairo, Egypt

       In an attempt to achieve water suitable for recycling from paper-mill effluent, photo-Fenton`s advanced oxidation and chemical coagulation were investigated as options to remove both non-biodegradable COD (chemical oxygen demand) and color from pre-bioremediated black liquor effluent. It was found that at the first bioremediation COD removal ranged from 35-98% and TSS (total suspended solids) removal ranged from 12-89%. Almost 20% of the heavy metals have been removed. Photo-Fenton`s/UV advanced oxidation and lime coagulation combination treatment achieved complete removal of COD, TSS and color. This was able to remove 50-100% COD, 47-100 %TSS and 30-100 % color. The removal of heavy metals was enhanced to reach>80% removal. Two potential microbial species out of the eight tested strains were the most dominant species in the three media. Diplodia oryzae and Phanerochaete chrysosporium NRRL6364 enhanced the post-physicochemical treatment to reach optimum clean up after 12 days of incubation at 32ºC. They produced a medium -grade recycled water. Operating costs are outlined.

 

 

2/17 OPTIMIZATION, PURIFICATION AND CHARACTERIZATION OF CERTAIN EXTRACELLULAR ACTINOMYCETAL LIPASE.

A.A. M. Abou Zeid; E.Y. Tohamy; N.M. Awny; Y.A.M. El Zawahry

and F.M. Reda.

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

Seven lipolytic active actinomycetes were isolated from cultivated soil samples in Dakahlia Governorate. The most potent isolate was identified as Streptomyces lipmanii. Maximum biomass, lipase activity and specific enzyme activity of St. lipmanii were obtained after six days incubation at 32oC in production medium adjusted at pH 7 under shaking condition. Optimum growth and lipase activity occurred in presence of 0.2% olive oil; 0.2% Arabic gum; 0.1% K2HPO4; 0.05% MgSO4.7H2O and 0.05% KCl in the production broth medium. Irradiation using low doses of gamma rays (0.25 – 0.75 kGy) had temporary initiating effect on viability and lipase production by St. lipmanii, after which gradual decreases in viable count and lipase activity occurred until the lethal dose (3kGy). Immobilized spores of St. lipmanii on artificial sponge cubes showed high microbial growth expressed as 7.4x106 cfu/ml and maximum enzyme activity (8.7U/ml) after 5 days incubation under shaking condition. Upon renewing the production medium every five days the immobilized lipase enzyme attained about 90%, 83%, 79% and 75% of its activity after third, fourth, fifth and sixth reaction cycles, respectively. Whereas fresh reaction mixture was renewed every five days (reaction cycle). Ammonium sulphate (70%) precipitation of cell free filtrate of St. lipmanii increased the specific enzyme activity (SEA) by 1.9 folds. Further purification by gel filtration using Sephadex G 50 and G100 increased the SEA by 5.5 and 104.05 folds, respectively. Analysis of purified enzyme using capillary electrophoresis revealed an apparent molecular weight of 45000 daltons after 3.39 minutes. Characterization of purified lipase enzyme showed that when 1ml pure enzyme was added to reaction mixture containing 12.5% emulsified olive oil adjusted at pH7 and incubated for 24 hours at 32oC, maximum lipase activity was obtained. Substituting olive oil in production medium by different concentrations of cotton seed litters waste revealed the capability of St. lipmanii to degrade lipids present in 8 gm waste/50 ml fermentation medium after 6 days incubation at 32oC under shaking condition.    

 

 

3/17 EVALUATION OF THE GENETIC BASE OF NINE SUGAR CANE VARIETIES USING PCR, PROTEIN PROFILE AND FLOWCYTOMETRY.

M.Z. Attallah; M. Osman*; O.M. Badawy** and H.A. Heikal*

Department of Chemistry and Physiology, Agricultural Research Station, Sabahia, Sugar Crops Research Institute, ARC, Alexandria, Egypt.

*Genetic Engineering and Biotechnology Institute, Minoufiya University.

**Department of Breeding and Genetics, Agricultural Research Station, Sabahia, Sugar Crops Research Institute, ARC, Alexandria, Egypt.

Comparative studies of nine sugar cane varieties were conducted using PCR, Protein profile and flowcytometry techniques. The sugar cane varieties were grouped on the basis of flowering as follows: the first group contained Co 464, Co 798 and Co 419 varieties not reached to full flowering. The second group included Co 534, Co 617 and Co. 453 not flowering varieties. The third group included Co 1155, Co 214 and Co 312 full flowering varieties. The results of PCR – RFLP- banding patterns revealed that there were relationship and similarity between the first group and the third group of sugar cane varieties. Regarding protein profile banding patterns, the results strengthened those obtained from PCR – RFLP DNA banding patterns. Comparing the data obtained from flowcytometry, it can be arranged according to the mitotic activity in the following rank group 3 >group 1 > group 2. However, this conclusion represents that the three groups, at the level of this analysis, displayed differential developmental stages.

 

 

4/17 PRODUCTION, PURIFICATION, MODIFICATION AND IMMOBILIZATION OF ALKALINE PROTEASE FROM BACILLUS STEAROTHERMOPHILUS

S.A. Ahmed; W.Gh. Shousha*; E.M.E. Mahdy*; H. Refaat* and M.A. Abdel-Naby

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

*Chemistry Department, Faculty of Science, Helwan University, Cairo, Egypt.

Six bacterial strains were tested for their abilities to produce active and thermostable alkaline protease. B. stearothermophilus was found to be the most potent alkaline protease producer (14.9 U/ml) and therefore it was used throughout this study. The effect of some cultural conditions on the productivity of alkaline protease by B. stearothermophilus was investigated. The strain used in its optimum medium afforded 88.36 U/ml of caseinase activity. The crude enzyme was optimally active at pH 9.5 and 50°C. Partial purification of alkaline protease was achieved by fractional precipitation of the crude enzyme by ethanol, ammonium sulfate and tannic acid. The ethanol fraction precipitated at 75% contributed 77.8% of the total recovered activity. This fraction was used for the succeeding part of the work (modification and immobilization of the enzyme). Some properties of the partially purified enzyme were investigated. Chemical modification of enzyme by covalent coupling of soluble polysaccharides has been reported as a common technique for improving its properties especially thermal stability. This method has been applied to alkaline protease using sodium periodate activated polysaccharides (pectin, amylopectin and dextran of different molecular weights). The enzyme coupled to activated pectin showed the highest thermal stability and retained the highest specific activity (917.1 U /mg protein). In general, the modification process of the enzyme protected it against heat inactivation. The enzyme was immobilized on insoluble supports by covalent binding. Of all tested carriers, the immobilized enzyme on nylon showed the highest immobilization yield (88.9%) and the highest immobilized activity (3112.2 U/gm). The properties of the free and immobilized enzyme were compared.

 

 

5/17 EVALUATION OF THE ANTIFUNGAL ACTIVITIES OF THE NOVEL SYNTHESIZED CHELATING CO-POLYMER EMULSION LATTICES AND THEIR SILVER COMPLEXES

A.L. Kansoh*, E.A.M. Youssef and M.A. Abd-El-Ghaffar

*Microbial Chemistry Dept. and Polymers & Pigments Dept.,

National Research Centre, Dokki, Cairo-Egypt

The novel binary chelating copolymers of butyl acrylate with itaconic and maleic acids were prepared by emulsion polymerization process. The chelating co-polymers of butyl acrylate-co-itaconic acid (BuA/IA) and butyl acrylate-co-maleic acid (BuA/MA) and their silver complexes were characterized and identified using IR and DSC measurements. The biological activities of these compounds were studied against various types of fungal strains. The dose and the rate of leached silver ions were controlled by the type of the co-polymers used and the solubility in the medium. The results provided laboratory support for the concept that the polymers containing chemically bound biocides were useful for controlling microbial growth. The silver uptake by fungal strains was studied to determine their difference in behavior to the antifungal activities of these compounds. The uptake strategy was examined by transmition electron microscope (TEM).

 

6/17 UTLIZATION OF BAKER’S YEAST TO PRODUCE YEAST AUTOLYSATES AND YEAST CELL WALL BIOPOLYMERS

(GLUCAN AND CHITIN)

A.M. Saad, H.M. Hassan and *M.A. Gadalla

Dept. of Microbial Chemistry and *Dept. of Microbial Biotechnology

National Research Centre, Dokki, Cairo, Egypt.

Baker’s yeast was grown on synthetic medium supplemented with sugar cane molasses (pH 5.5) in a 3 L New Brunswick bioreactor at 28ºC. The growth rate of yeast increased by the increasing of oxygen consumption until reached to maximum after 14 h (oxygen consumed, 76.7 %). Treatment of intact yeast cells with different types of buffers with stirring at 50 ºC for 20 h markedly increased the release of proteins (48 %). This procedure has been used for the commercial production of yeast autolysate. The produced yeast autolysate contained considerable amounts of the B – vitamins in addition to niacin and trace mineral elements and phosphorous (7.65 – 8.49 %). The proximate amino acid composition of produced autolysed yeast powders was determined and amino acids includes 17 amino acid. A more useful index of more flavour potential is indicated by the free amino acid contents in the extracts. Another yeast component with growing importance is glucan which was separated by the treatment of yeast cell wall by 10 % NaOH. The alkali insoluble extracted residues obtained amounted to 15 % of the original cell wall material which contained a small amount of chitin. Chitin was isolated from chitin – glucan complex by treatment with cold conc. HCl. The lyophilized product was 0.1 % of the isolated compound. The residue obtained after the solubilization of chitin by acid treatment was characterized as being mostly glucan. The isolated biopolymers were identified by IR spectrophotometry and it were correspondent with those obtained from Sigma Co.        

7/17 MICROSCALE TESTING OF SOME PESTICIDES IN WATER BY MICROBIAL DEGRADATION

M.M. Hazaa

Batony Department, Faculty of Science, Zagazig University, Benha

A relatively simple, aerobic and anaerobic biodegradation test method for pesticides was proposed. The biodegradation procedure steps include:   inoculation (embding) substance (pesticide) under investigation to a substrate of non-acclimated microbes such as activated sludge, field soil and river sediments in stirred flasks. This method was applied to evaluate the biodegradability of twelve pesticides. Obtained biodegradation rate constants showed a parabolic correlation with the 1-octanol-water partition coefficients of the pesticides and a linear one with their alkaline hydrolysis rate constant. Therefore, the biodegradability of pesticides could be used predicted as an indicator for the partitioning solvent from the aqueous medium to the membrane of organismes by these two parameters.

8/17 NON-INVASIVE IMMUNOASSAY FOR THE DETECTION OF HELICOBACTER PYLORI INFECTION IN EGYPTIAN PATIENTS

Y.A. El-Zawahry, M.F. Ghaly, G.G. Ibrahim*, A. Abdel-Mageed* and G. Balah*

Botany Department, Faculty of Science, Zagazig University

*Biotechnology Research Center, New Damietta, Egypt

       A bacterial isolate identified as Helicobacter pylori (H. pylori) was isolated from a gastric biopsy of an Egyptian patients with symptomatic and signs suggesting gastritis. The total protein content of the bacterial lysate of H. pylori was 2mg/ml. The protein pattern of isolated H. pylori was resolved using 12% Sodium Dodecyl Sulphate –Polyacrylamide Gel Electrophorsis: (SDS-PAGE) showed 18 bands. The molecular weights of protein bands ranged between 180 to 20 KDa in descending order. Sera of 120 individuals (82 males and 38 females) showing dyspeptic symptoms and 32 asymptomatic healthy volunteers (18 males and 14 females) were screened for IgG antibodies using ELISA test. The antigenic properties of blood sera of 28 individuals; 16 showing positive H. pylori infection and 12 non-infected, were investigated using indirect ELISA against H. pylori previously prepared from bacterial lysate.   Incidence of positive ELISA test was 91.7 % (110 cases) in the symptomatic individuals and 25 % (8 cases) in asymptomatic individuals.. No relation has been shown between H. pylori infection and age. The prevalence of H. pylori infection is the same among males and females. It could be concluded that the use of serological test and ELISA will be useful as a screening test for H. pylori infection without the need of endoscopy.

 

9/17 HIGH – LEVEL XYLANASE PRODUCTION BY RADIO- RESISTANT, THERMOPHILIC BACILLUS MEGATERIUM AND ITS MUTANTS IN SOLID – STATE FERMENTATION.

M.A.M. Abo-State

Department of Microbiology, National Center for Radiation Research and Technology, Nasr City, Cairo, Egypt.

Bacterial xyalanse (EC. 3.2.1.8) can be produced from cheap agro – industerial waste wheat bran (WB) by radioresistant, thermophilic isolate of B.megaterium in solid state frementation (SSF). The highest xylanase activity was found at mositure 1: 2.5 (Solid: dis.water) ratio, pH 6.0 after 48 hours incubation at 50°C, inoculated with 15 ml bacterial culture,   when the enzyme was extracted by sodium acetate buffer (pH 5.5). Exposure of B. megaterium to 15 kGy gamma irradiation, reduced, the viability by 6.3 log cycles. Forty six mutants had been isolated. Mutant No 36 (M10K) exposed to 10 KGy produced the highest extracellular protein and xylanase activity (700 mg/ml and 9993 U/g). This mutant (M10K) when exposed to UV – irradiation, the viability was reduced by 3.9 log cycles after 15 min. The hyperproducing xylanase were mutants exposed to 4 and 15 min. (M10K-4 and M10K-15). Under the optimized conditions, the highest xylanase activity was 10,350 U/g produced by mutant M10K-4.

 

 

10/17 INORGANIC PHOSPHATE SOLUBILIZATION BY FREE OR IMMOBILIZED TRICHODERMA HARZIANUM CELLS IN COMPARISON WITH SOME OTHER SOIL FUNGI

M.S. El-Katatny

Botany Department, Faculty of Science, El-Minia University 61519, Egypt

In this study the ability of T. harzianum Rifai (T24) to solubilize and release phosphate from tricalcium phosphate in comparison with some other soil fungi was tested. The different tested fungi varied in their P-solubilization efficiency (E) on agar plates. Free mycelia of A. niger, T. harzianum, P. italicum and P. duclauxi were further tested for their abilities to solubilize tricalcium phosphate in Czapeks liquid medium. Addition of tricalcium phosphate resulted in increase of fungal growth. Although T. harzianum had no solubilization activity in plate assay, it showed a relatively good P-solubilization activity in liquid culture. Immobilization has generally improved the fungal ability to solubilize P in the medium as compared with fungal free mycelia. Beads entrapping fungal spores were successfully used for P-solubilization for 7 repeated times in presence of fresh medium supplemented with tricalcium phosphate. In conclusion, encapsulated formulations of the biocontrol agent T. harazianum are suitable mean for phosphate solubilization which could add to improvement of plant growth under conditions of phosphate insolubility.

 

11/17 ANATOMICAL STUDIES ON SOME CROTALARIA L. (FABACEAE) COMPARATIVE STUDY

A.A. Khafagy; A.H. Mohamed; M.A. El-Kholy and A.T. Salama

Botany Department, Faculty of Science, Al-Azhar University    

The anatomical attributes of stems and leaves were studied to evaluate their significance in differentiation between five species of Crotalaria (Fabaceae). The anatomical features appeared to be of significant importance in differentiation between studied taxa. The rnorphology ofboth the lowerand upper leaf surfaces in the Crotalaria species has been investigated using scanning electron microscope (SEM). Artificial key based on such characters was designed.

 

 

12/17 A CONTRIBUTION TO THE TAXONOMY OF THREE LEGUMINOUS TREES. [EVIDENCE FROM RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) ANALYSIS]

A.A. Shehata, M. Osman* and M.H.A. Loutfy**

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

*Genetic Engineering & Biotechnology Research Institute (GEBRI), Sadat City, Minufiya University, Egypt.

**Biological Science and Geology Department Faculty of Education, Ain Shams University, Cairo, Egypt.

Random amplified polymorphic DNA (RAPD) analysis was used to study the relations between Caesalpinia gilliesii, C. pulcherrima and Delonix regia (Caesalpinoideae- Leguminosae) using three deca-mer primers. Compiled data from the literature on macromorphology, SEM of seed coat, embryology and palynology were used in addition to the present results of RAPD analysis and analyzed by the NT Sys-PC program for UPGMA analysis.   The obtained results showed a close similarity between Caesalpinia pulcherrima and Delonix regia.   The primer OPA-12 was shown to produce the highest number of the species specific bands and so can be recommended for differentiating the studied taxa.

 

 

13/17 ECO-PHYSIOLOGICAL AND PHYTOCHEMICAL EVALUATION OF THREE MESEMBRYANTHEMUM SPECIES

H. E1-Tantawy, F. E1-Shayeb and A. E1-Kholy*

Botany Department, Faculty of Science, Menoufiya University,

Shebin E1-Koum, Egypt.

* Desert Research Center, Mataria, Cairo.

Three species of Mesembryanthemum viz M. crystallinum, M. nodiflorum and M. forsskalei were evaluated for their ecophysiological behavior and phytochemical characteristics. Succulent, pigment(s) content, ash, mineral and secondary product contents were determined. The three species of Mesembryanthemum were characterized by the highest value of succulent and limited fluctuation in their pigment(s), mineral ions and ash. The phytochemical screening showed that the M. crystallinum L. (edible wild plant) are rich in   lipids (8.512%) and moderate content of protein (4.90%), while M. forsskalei Hochst. (Seeds are edible) are poor in lipid (1.849%) with moderate content of protein (3.2%). The three investigated species are very poor in carbohydrate contents.Free and combined sugars, rhamnose, arabinose, ribose, mannose and fructose were detected in the three species of Mesembryanthemum. The highest percentage of total protein amino acids was found in M. crystal/num (14.53) followed by M. nodiflorum (3.90) and M. forsskalei (3.01). Most of the essential amino acids were detected in the three species. The fatty acids namely dodecyclic, tridecyclic, pentadecyclic, palmetic and stearic were detected in all species. several fatty acids ( palmetic, stearic, oleic and linoleic) were   detected in the investigated species. Two flavonoids were isolated and identified, as apigenin 7 glucoside and vitexin.

 

14/17 DETECTION AND CYTOPATHIC EFFECT OF POTATO SPINDLE TUBER VIROID INFECTING POTATO PLANTS

Kh.A.; El-Dougdoug, M.H. Abdel-Ghaffar; R.M. Taha* and M.M. Hazaa**

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

* Department of Botany, Faculty of Science, Cairo University

(El-Fayoum Branch), El-Fayoum, Egypt.

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

(Benha Branch), Benha, Egypt.

Potato spindle tuber viroid (PSTVd) was detected in potato tubers exhibited PSTVd-like symptoms using biological and molecular techniques. The results of mechanically inoculated tomato (Lycopersicon esculentum cv. Castle Rock) and Scapolia sinensis plants with clarified extracts of tuber sprouts exhibited PSTVd-like symptoms showed PSTVd-characteristic symptoms. A RNA with structural properties typical for PSTVd-RNA was detected in total nucleic acids using return-polyacrylamide gel electrophoresis (R-PAGE). Using reverse transcription polymerase chain reaction (RT-PCR) and specific primers for PSTVd, a major of cDNA product of approximately 360 bp (full length) was detected in total nucleic acids extracted from PSTVd-infected tissues. The products of RT-PCR were confirmed using Southern blot hybridization after hybridized with non-radioactive, digoxigenin (DIG)-labeled PSTVd-cRNA probe and visualized by chemiluminescent detection. No hybridization was obtained between the probe and RT-PCR products from healthy tissues. Ultrastructure of the cells of PSTVd-infected potato leaves showed two important variations from cells of healthy leaves. First, paramural bodies known as plasmalemmsomes and watchspring-like multilayered membranes with different sizes and shapes were showed in cytoplasm of infected cells. Second, aberrations of the thylakoid membrane system of the chloroplasts, and the grana were not well developed in PSTVd-infected cells.

 

 

15/17 EFFECTS OF GREEN, RED AND BLACK TEA BLEND ON LEVELS OF EXPRESSION OF GP-X AND THE CONCOMITANT MODULATORY EFFECTS ON CYP11B2 EXPRESSION IN MICE LIVER CELLS

H.A.S. Oraby

Cell Biology Department, National Research Centre, Egypt

Tea (Camellia sinensis) is one of the most widely consumed beverage in the world. The role of tea on the transcriptional regulation levels of GP-x mRNA and its related modulator effects on CYP11B2 expression in mice liver cells were investigated, in the present work, using the RT-PCR assay. Glutathione peroxidase (GPx) is one of the principle antioxidant and detoxifying enzymes in the cell. The aldosterone synthase gene (CYP11B2) is considered as a strong stimulator of fibrinogenesis and mitogenesis. Drinking a 1% mixture of green, red and black tea for two weeks, as a sole source of drinking, significantly up regulated the expression of GP-x in mice injected with a single dose of cyclophosphamide, as a mutagen, 24 hours before being sacrificed. A concomitant down regulation of CYP11B2 was also recorded in association with the treatment. The ability of the green, red and black tea mixture to modulate the expression of GP-x and CYP11B2 may be adopted as the underlying strategy to prevent mutagenicity and the subsequent carcinogenicity associated with the treatment with cyclophosphamide.

16/17 MORPHOLOGICAL DIVERSITY OF DATE PALM (PHOENIX DACTYLIFERA L.) IN EGYPT

II-SEMI-DRY DATE CULTIVARS

R.M. Rizk and S.S.H. El Sharabasy*

National Gene Bank (NGB), *The Central Laboratory of Date Palm Research & Development, Agricultural Research Center, Giza, Egypt.

Taxonomical relationships of ten semi-dry date cultivars growing in Egypt were addressed based on one hundred morphological attributes of trunk, crown, leaves, fruits and seeds. The most important characters are arranged according to their taxonomic significance. This is followed by a key to the semi-dry date cultivars in Egypt. Fruit and seed attributes are still the most important criteria to distinguish among date palm cultivars. The study concluded that there is a window of opportunity to conserve semi-dry date cultivars. Positive conservation action may be necessary at the infraspecific level if diversity of date palm cultivars are to be maintained. The active cultivation is vital to survival and a cultivar is soon lost for ever if it is not regularly propagated.

 

 

17/17 USE OF PLANT GENOTOXICITY BIOASSAY FOR THE EVALUATION OF EFFICIENCY OF ALGAL BIOFILTERS IN BIOREMEDIATION OF TOXIC INDUSTRIAL EFFLUENT

H.M. Abdel Migid*, Y.A. Azab and W.M. Ibrahim

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

The toxicity and efficacy of an algal-based bioremediation technology were assessed through bioassays for ecological risk of contaminated industrial effluents. The algal bioremoval of heavy metals was evaluated using an in vitro approach.   Phytogenotoxicity tests were conducted with Allium cepa and Vicia faba plants to evaluate the genotoxicity of the industrial effluents before and after treatment with different kinds of algal biofilters. Root cells were exposed for 24h to different dilutions of both raw and treated effluent of a chemical fertilizers factory. Three cytogenetic endpoints were used to assess the mutagenic potencies of the industrial effluent: mitotic inhibition, mitotic chromosome aberrations, and nuclear irregularities in interphase cells.   Before algal treatment, the industrial effluent caused strong genotoxic effects represented by severe inhibition in mitotic activity of meristematic cells and high frequency of both chromosome and nucleus abnormalities. After algal treatment, the cytotoxic effect of 30% and 60% concentrations of the treated effluent was comparable to that of 5% and 10% concentrations before treatment, respectively and the frequency of both chromosome and nuclear abnormalities declined up to approximately 50%. Statistical analysis of the results indicates a significant reduction in genotoxicity after bioremediation that pointed out by a remarkable reduction in heavy metal concentrations. The Allium and Vicia genotoxicity approach was effective in monitoring bioremediated effluent for toxicity.

 

 

18/17 ENHANCEMENT OF YEAST SACCHAROMYCES CEREVISIAE UNSATURATED FATTY ACID CONTENT VIA PROTOPLAST FUSION BETWEEN THE SAME MATING TYPE

Sh. Sharaf El-Deen and Y.M. Ahmed

Microbial. Genetics Dept. and Microbial. Biotech. Dept. National Research Centre, Cairo, Egypt

Preliminary results for the production of unsaturated fatty acid (USFAs) were obtained. The comparison between two mating types (α and a) of S. cerevisiae with the indicated that the production capacity is correlated by (a) mating type. In order to improve USFAs content specially ω-3. Diploid construction is difficult or impossible in natural way between the same mating type. Protoplast fusion only is the technique that could be used for such illegitimate copulation. It is an important to collect integrate desirable genes from different haploid strains with the same mating type into diploid one. Complementation between strains on selective medium was used to detect and construct the diploids phase. Phloxin-B stain can differentiate between haploids, diploids and dead colonies. Auxotrophic mutants of the same mating type are visualized by different copper resistant as visual pigment with their requiring of amino acids and bases The fatty acid methyl esters (FAMEs) were analyzed by GC/MS . The productivity of the two fusants are varied greatly in USFAs ( ω-3). The first fusant (GT/IL) produced 134.15% as its parental strain (IL-126-7A) and, LBC/IL the second fusant produced 638.38 % as its the same parent. The second fusant produced 475.85 % as its first fusant (GT/IL).The results obtained confirmed the importance of protoplast fusion followed by selection as an important breeding tool in yeast for improvement of unsaturated fatty acid production. The highest productive of fusant (LBC/IL)was less asi constructed 79.27% as its first fusant (GT/IL) and containing long chain fatty acids within scan area (3858) in sheet GC/MS.

19/17 STUDY OF THE INFLUENCE OF ELECTRIC FIELD EXPOSURE ON SOME SOIL MICROBIAL ACTIVITIES

W.A. Hassanein and A.A. Ali

Department of Botany, Faculty of Science, Zagazig University

In this study, the focus on response of soil microbes to electric field (EF) of different exposure periods was detected. Exposure of different soil samples to electric field strength of 6KV/m for 1,2 and 3 months slighty decreased the total viable counts of general bacteria, as well as Azotobacter and Azospirellum, however, the total viable counts of fungi reflected noticeable decrease compared to bacteria. Data of unexposed soil showed a significant increase in CTMB (total microbial biomass C), CAMB (active microbial biomass C), qR (metabolic quotients), BR (basal respiration), while significant decrease in qCO2 at the end of incubation period was observed. Also significant decrease in CTMB, CAMB and BR of soil under the effect of three months exposure to electric field was indicated. Soil enzyme activities decreased by increasing exposure period up to 3 months. Soil nitrogenase was more sensitive to EF followed by; acidic phosphatase, urease, dehydrogenase and alkaline phosphatase, they decreased by 30, 23.6, 20, 13 and 7.7% respectively compared to the unexposed samples after the same exposure period. Growth rate of the selected Azotobacter isolate indicated significant decrease in the viable cell counts when the bacteria were grown in liquid medium and exposed to the electric field for 1,2,3,4,5 and 6 days compared to unexposed ones. Regarding to the electrophoresis analysis of extracted protein, each of exposed and unexposed Azotobacter cultures incubated for 3 and 6 days had 14 protein bands. Two bands (Mw83.4 and 15.9KDa) were disappeared, and two others bands (Mw 86.3 and 16.2 KDa) were appeared after exposure for 3 and 6 days. The average optical density of bands decreased at exposure period 6 days compared to the unexposed ones. This study concluded that the action of EF causes an intensified slow down of either growth and activities of soil microorganisms.

20/17 SUPPRESSION OF MITOTIC PROCESS WAS ASSOCIATED WITH METAPHASE ARREST AND CHANGES IN ULTRASTUCTURAL PATTERN OF ALLIUM CEPA L. ROOTS:

I. ROSMARINUS OFFICINALIS L.WATER EXTRACT SUPPRESSION OF MITOTIC PROCESS WITH METAPHASE ARREST

S.A.F. Tawab, Z.M. Adam and Sh. Selmi

Botany Department,Faculty of Girls, Ain Shams University

Experiments were carried out to investigate the potential for using extract from Rosmarinus officinalis L. as a mitostatic and metaphase arrest agent. Allium cepa L. meristem was used as a biological system in this investigation. Different concentrations were made to establish the optimum dose response. The drug was concentration- dependent. Metaphase arrest with complete inhibition in spindle assembly was common feature. The metaphase to anaphase transition was blocked leading to discontinuous mitotic activity. Meanwhile, the drastic inhibition in protein synthesis in spite of accumulation of protein synthesis devices investigated by electron microscopy, gesticulate the mode of action of the drug which has been discussed.

 

21/17 THE MUTAGENIC EFFECTS OF THE BIOPESTICIDE AGRINE ON VICIA FABA PLANT

N.M. George, E. Abd El-Haliem and A.A. Bolbol

Botany Department, Faculty of Science, Zagazig University

The effects of biopesticide Agrine on mitosis, meiosis, yield and seed protein electrophoresis of Vicia faba plant were studied. A decrease in the mitotic index in the root tip meristems of Vicia faba was observed after seed soaked in half recommended (1/2RD), recommended (1RD) and double recommended (2RD) doses of Agrine for 12, 24 and 48 hours. Agrine also induce a range of chromosomal irregularities, their frequencies were dose dependent. The dominant abnormalities induced in dividing cells were chromosomal stickiness and disturbed phases. Agrine has the ability to exert an action on the different stages of meiotic divisions I and II. The chromosomal abnormalities of meiotic division were nearly similar to the corresponding in mitotic division but their frequency was higher. The M1 progeny showed a marked percentage of abnormal cells and reduction in the productivity. These abnormalities were similar to those of parent plants. These results indicate the toxicity of Agrine and its mutagenic potential. The mutagenic effects of Agrine on banding patterns of protein showed many changes such as changes in band intensity, changes in band mobility, appearance of new bands and disappearance of some others.

 

22/17 A STUDY ON BIODESULFRIZATION OF AN EGYPTIAN CRUDE OIL

I- EVALUATION OF BIODESULFRIZATION CAPACITY OF RHODOCOCCUS RHODOCHROUS ATCC 53968

M.A. Salama, M.K. Ibrahim* and M.R. El-Shahawy

Petrochemicals Department, Petroleum Research Institute, Nasr City, Cairo, Egypt

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

The capacity of the mutant strain Rhodococcus rhodochrousATCC 53968 to desulfurize an Egyptian crude oil (Balaeem Barry) was evaluated. The optimized parameters affecting the biodesulfurization process were incubation for 24 hours at 30°C with pH adjusted to 7.2 and the best oil concentration was 1.0 g /100 mLmedium. Meanwhile, glucose (10.0 g / L medium) and ammonium chloride (3.0 g / L medium) were best favored as carbon and nitrogen sources respectively. γ-Irradiation of the bacterial strain seemed to inhibit the sulfur biodegradation enzyme system. The biodesulfurized crude oil indicated a pronounced drop in sulfur content in asphaltenes while no difference was monitored for saturates, aromatics and resins. Microbial desulfurization of Balaeem crude oil was associated with a decrease in paraffins and isoparaffins bearing 13-18 carbon atoms and an increase in those having 21-30 carbon atoms. Slight degradation of the biomarker phytane was observed.

 

23/17 A STUDY ON BIODESULFURIZATION OF AN EGYPTIAN CRUDE OIL

II- EVALUATION OF BIODESULFURIZATION CAPACITY OF SOME LOCAL BACTERIAL ISOLATES

Mohamed K. Ibrahim, Mohamed A. Salama*, Thanaa Abdel-Moghny** and Mohamed R. El-Shahawy*

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

*Petrochemicals Department, Egyptian Petroleum Research Institute,

Nasr City, Cairo, Egypt

**Petrochemicals Applications Department, Egyptian Petroleum Research Institute,

Nasr City, Cairo, Egypt

Out of six local bacterial isolates, the isolate caused the maximum desulfurization of an Egyptian crude oil (Balaeem Barry) was an isolate identified as Bacillus subtilis. It was found that the optimum conditions for desulfurization were 30ºC incubation temperature for 24hrs, pH 8.0 and oil concentration 1 g/100 mL medium. Also, glucose (10 g/L medium) and ammonium chloride (3 g/L medium) were the best carbon and nitrogen sources, respectively. g-Irradiation seemed to inhibit the biodegradation enzyme system. A comparative study regarding reduction in sulfur content and alterations in hydrocarbons constitution of crude oil asheld using this local isolate and that took place using the mutant strain Rhodococcus rhodochrous ATCC 53968 studied in a previous work. B. subtilis has a distinct biodesulfurization capacity when taking in consideration its ability to lower the aromaticity of crude oil and increase paraffinic and isoparaffinic contents.


24/17 Studies in the Leaf Anatomy of some Species of the Arecaceae.

I- Subfamily Coryphoideae

E.A.K. Karakish and S.A. Hassan

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

The leaf anatomy of 17 species belonging to subfamily Choryphoideae (Arecaceae) is investigated. The present work refers to a group of characters (seven characters) which are shared by all the studied species. A second group of other seven characters are also shared by 11-14 species. A third group of characters are found to be exclusive for Chamaerops humilis L., Latania lontaroides, Licuala grandis, Livistona chinensis, Washingtonia filifera and Washingtonia robusta. On the bases of investigated characters, the species are categorized into two groups, one of them contains two subgroups.

 

 

25/17 OPTIMIZATION OF INDUSTRIAL PRODUCTION OF RIFAMYCIN B BY AMYCOLATOPSIS MEDITERRANEI

IV. PRODUCTION IN THE FERMENTOR

O.M. El-Tayeb; M.M.M. Hussein*; A.A. Salama* and H.F. El-Sedawy

Microbial Biotechnology Center and the *Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt.

Optimization of the physical and physiological parameters of the fermentation process using the gene amplified variant of Amycolatopsis mediterranei (NCH) was carried out using the fermentor. Optimization of the physical parameters by controlling the pH at 6.5 for 3 days then at 7 thereafter and by adjustment of aeration at 1 vvm for 3 days then controlling the dissolved oxygen (DO) at 30% saturation   increased the yield from 9.77 to 11.96 (22%) and 13.39 g/l (37%), respectively. Replacing 12% glucose in the fermentation medium (F2m1) with 5% glucose syrup (F2m3 medium) resulted in a drop of the yield from 9.77 to 7.5 g/l, while further addition of another 5% glucose syrup at day 4 increased the yield from 7.5 to 13.81 g/l (84 %); with a further increase in the yield to 14.25 g/l (90%) upon controlling DO. Whereas, the combined addition of 0.1% yeast extract at day 2 to F2m3 medium along with the addition of 5% glucose syrup at day 4 increased the yield from 7.5 to 15.35 g/l (105%); with a further increase in the yield to 16.3 g/l (117%) upon controlling DO. The fed-batch addition of both 3% soytone at day 3 and 5% of glucose syrup at day 4 to F2m3 medium increased the yield from 7.5 to 16.2 g/l (116%) and by extending the fermentation period to 10 days the yield reached 17.9 g/l (139%). Upon applying all optimum physical and physiological conditions in the fermentor the yield increased from 7.5 to 17.43 g/l in 8 days (132%) and by extending the fermentation period to 10 days the yield reached 19.4 g/l (159%). Further process optimization by examination and analysis of the kinetics of the process would most certainly further increase the yield and quantitatively define the process to a level that could be tested on a pilot scale.

 


26/17 GENETIC TOXICOLOGY OF NABU-S HERBICIDE ON CHROMOSOMAL BEHAVIOUR, CHIASMA FREQUENCY, POLLEN FERTILITY AND SEED STORAGE PROTEIN

OF VICIA FABA

E. Abd El-Haliem

Botany Department, Faculty of Science, Zagazig University

The genotoxic effect of Nabu-S herbicide in Vicia faba plant was investigated using chromosomal behaviour in somatic and gametic cells, chiasma frequency, pollen sterility and the alterations in the M1 seed storage protein banding. The used herbicide doses had a mitodepressive effect and induced a highly significant percentage of mitotic and meiotic anomalies. For meiotic studies, three treatments were used. These were the treated dry seeds as pre-emergence, treated seedlings and flower buds as post-emergence. Mutagenic potential of the herbicide resulting in an action on chromosomes such as the most dominate chromosomal stickness and chromosomal fragmentation in addition to a disruptive action on spindle apparatus which were C-metaphase, C-anaphase, chromosomal disturbance, lagging chromosomes, tripolar and multipolar ana-telophase. The karyotoxic activity of Nabu-S in meiosis showed a high efficiency in the reduction of chiasma frequency per PMC. Moreover, herbicide treatments induced pollen sterility significantly. The degree of which increase with increasing herbicide dose. At the protein level, Nabu-S herbicide showed mutagenic effects in the patterns of the M1 seed storage protein banding of Vicia faba. In general, these chromosomal and genetic damages indicates a genotoxic and mutagenic potential of a new Nabu-S herbicide in Vicia faba.

 

27/17 PURIFICATION AND CHARACTERIZATION OF TWO POLYGALACTURONASES PRODUCED BY

ASPERGILLUS FUMIGATUS

A.A. Salama, M. A. Ramadan, A. M. Hashem and S.M. Mohammady

Department of Microbiology and Immunology, Faculty of Pharmacy,

Cairo University

       A total of 100 bacterial and fungal isolates having pectinolytic activity were recovered from 150 samples of soil and rotten vegetables and fruits. Fungal isolates were found to be much higher producers than bacterial isolates. They produced different enzymes Polygalacturonase, Polygalacturonate lyase, Polymethyl-galacturonase, Polymethylgalacturonate lyase, and Polymethylgalacturonate esterase. Different physical and physiological factors affecting the production of Polygalacturonase produced by the five most active fungal isolates were studied including temperature, time of incubation, shaking, carbon and nitrogen sources. Two exopolygalacturonases (exoPG-I and exoPG-II) were produced by Aspergillus fumigatus. The two enzymes were purified by gel filtration on sephadex and ion exchange diethylaminoethyl cellulose. The exoPG-I and exoPG-II were purified 19.3 and 21.5 fold, respectively. The two enzymes have the same molecular weight of 78.75 KD. They have similar bimodal pH profiles with two pH optima at 5.6 and 8.3. Their optimum temperatures were 45oC and 55oC, respectively. Complete inactivation of the two enzymes was observed with NH4+, Zn2+, Hg2+, Cu2+, Cr2+, Ca2+ and Al3+. Variable degrees of inactivation were observed with other metallic ions: Ni2+, Ba2+, Mg2+, Fe3+, Na+, Li+, K+ and Mn2+. On the other hand these enzymes were stimulated by Fe2+. PG-I and PG-II hydrolyzed polygalacturonic acid and to a lesser extent citrus pectin. The mode of action of the two enzymes was revealed by viscosimetric and reductometric methods and confirmed by thin layer chromatography of substrate hydrolysates.

 

28/17 PRODUCTION, PHYSICO-CHEMICAL PROPERTIES AND BIOLOGICAL ACTIVITIES OF AN ANTIBACTERIAL AGENT PRODUCED BY STREPTOMYCES FRADIAE

H.M. Atta; M.E. Zain and A.T. Abul-Hamd

Botany and Microbiology Dept., Faculty of Science, Al-Azhar University,

Cairo, Egypt.

An actinomycete culture was isolated from a soil sample collected from El-Menoufiya governorate, Egypt. This isolate (AZ-M-54) was found to be active against gram- positive and gram-negative bacteria. From the taxonomic features, the isolate matches with Streptomyces fradiae in the morphological, physiological and biochemical characters. Thus, it was given the suggested name Streptomyces fradiae. The active metabolites were extracted by chloroform. The separation of the active ingredient and its purification was performed using thin layer chromatography (TLC), column chromatography and bioautography techniques. The physico-chemical characteristics of the purified compound including color, solubility, melting point, elemental analysis, spectroscopic characteristics and chemical reactions have been investigated. The biological activities (MICs), and effect on protein and nucleic acid of the antibiotic were determined. The collected data emphasized the fact that the antibacterial agent, Clarithromycin, was suggestive of being related to Macrolide antibiotics produced by Streptomyces fradiae.

 

 

29/17 PRODUCTION, PROPERTIES AND IN VITRO APPLICATION OF NOVEL FIBRINOLYTIC ENZYME BY

BACILLUS MACERANS 3185

A.S. Ismial, S.A.S. Emam* and N.M.A. El-Shayeb

Department of Chemistry of Natural and Microbial Products & *Department of Microbial Biotechnology, National Research Centre, Dokki, Cairo, Egypt

       The two local isolates Bacillus macerans 314 and 3185, were studied in different media for the production of highly active fibrinolytic enzymes. Bacillus macerans 3185 in 2 days shaken culture composed of alkali-extracted soybean (as N source) and 0.2% (w/v) starch afforded the most highest activity of fibrinolytic enzyme. The crude lyophilized enzyme showed its optimum activity at 37 oC after 30min reaction and pH 6.8. The suitable substrate and enzyme concentrations were 16 µg and 2.4 mg per ml, respectively. Applying the specified optimum conditions, the crude lyophilized enzyme showed high efficacy for human blood clot resolution in vitro, wherethe complete clot resolution was recorded after 45 min.

 


30/17 TREATMENT OF OILY WASTE WATER USING COAGULANTS

G. El Diwani, S. Hawash, Sh. El-Rafie and N. Attia

Department of Chemical Engineering, National Research Center, Cairo Egypt

The aim of the study is the comparison between using polyelectrolytes and bitterns in treating oily sludge wastewater. The polyethanol amine (PEA) and   Hydrophobically associated cationic polymer (HACP) prepared through this work and imported (zetag -92) were used in the treatment. Polyelectrolytes have been used to enhance removal of suspended solids by sedimentation. The addition of some inorganic coagulants like ferric chloride is needed to improve the treatment process. Using FeCl3 combined with PEA or HACP, at optimum conditions, reduced turbidity, suspended solids and COD in range of 96.3 – 97.8 %, 96 – 100 % and 70 -100 % respectively. When PEA was combined with FeCl3 the settling rate was slightly improved and similar results were obtained using FeCl3 or LB singly after 5 minutes settling time. This investigation was conducted on samples from car wash   plant`s effluent.

 

 

31/17 CIRCULATION OF QUARON LAKE WASTES

II-GROWTH OF SCENEDESMUS SP. UNDER MG RESIDENCES

A.B. El-Sayed

Plant Nutrition Department, National Research Centre

A residence from an Egyptian salt production company rich in Mg content was used as supplementary nutrient to the growth medium of the isolated green alga Scenedesmus sp. Besides the higher Mg+2 content, sulfate, carbonate and bi-carbonate represented the accompanied cations by the equivalent amount of Mg+2. The first series of the experimental trial including 2, 4, 6, 8 and 10ml.l-1 of Mg solution, while they extended within the second series by 10 m.l-1 over each previously mentioned treatment. Third series including 10, 20, 30, 40 and 50 m.l-1. Data showed that with the advantage age, cell dry weight was proportionally increased as a result of residence increments during the first series (0-10 ml.l-1 Mg residences). During the second series (0-20 ml.l-1 Mg residences), more enhancements were observed. During the third series, slight decline was observed with super Mg residences including the volumes of 30, 40 and 50 ml.l-1 Mg residences. Chlorophyll accumulation showed a similar trend of dry weight except with those obtained with super Mg residences. A slight decline on chlorophyll accumulation was observed with 40 and 50 ml.l-1Mg residences. Thus, Mg residences could be successfully utilized as a commercial growth medium for-semi saline algae on the expense of such high priced solutions used for mass production.

 

 

32/17 PHYSICAL AND CHEMICAL STUDIES ON SOME RAPESEED VARIETIES GROWN UNDR DIFFERENT LEVELS OF NITROGEN FERTILIZATION.

M.H. El-Kholy; M. M. El-Zeky, Sh.Z. Saleh and S.G. Metwaly

Soils, Water and Environment Research Institute, Arc, Giza, Egypt.

The present work was carried out at El-Serw Agric. Res. Station during the two successive seasons 2002/2003 and 2003/2004, to study the effect of four levels of nitrogen fertilization (0, 30, 60 and 90 kg/N/Fed) on three rapeseed varieties, namely, Pactol, Serw 4 and Serw 6. The study included seed yield and some yield components, some physical and chemical properties of the extracted oils and total glucosinolates of the seed. The obtained results could be summarized as follow: 1- Serw 6 var. gave the highest seed yield followed by Serw 4 and Pactol, which gave the lowest yield. 2- the optimum N dose for realizing the highest seed yield for Serw 4 and Serw 6 was 90 kg N/Fed while, for Pactol was 60 kg N/Fed. 3- Pactol Var. contained the highest content of oil compared with the other two varieties, but it gave the lowest value of seed yield and subsequently oil yield. Increasing N rate up to 90 kg/Fed decreased seed oil content, but increased oil yield/Fed and protein content and protein yield in all varieties. 4- Negligible differences in acid, peroxide and saponification values were occured. While, refractive index and iodine values of oils of Serw 4 and Serw 6 were similar and exceeded those of Pactol. N fertilization had no effect on acid, peroxide and saponification values, while N rates up to 90 kg/Fed caused a slight decrease in iodine value and refractive index. 5- Total saturated fatty acids were similar in the three varieties and ranged between 6.2 and 7.8%. Palmitic was the most predominant. 6- Oleic was the predominant unsaturated fatty acid in the three varieties. 7- The highest amount of linoleic acid was found in Serw 4 followed by Serw 6 and Pactol. So, Serw 4 characterized with high content of essential fatty acid linoleic indicating high nutritive value. 8- Erucic acid content of the three rapeseed oils was very low to be, less than 2% of fat. Therefore, there is no hazard in consumption of safe rapeseed oil extracted from the varieties under investigation. 9- Nitrogen rates up to 90 kg/Fed caused little differences in fatty acid composition. For instance, oleic acid content increased at the expense of linoleic and linolenic acids. 10- All rapeseed varieties under study characterized by having low contents of glucosinolates (less than 30 μmol/g meal), N fertilization had insignificant effect on total glucosinolates in meals of rapeseed varieties. 11- Finally, increasing N rates increased NPK contents in seeds of the three rapeseed varieties.      

 

 

33/17 STIMULATION OF CELL DIVISION AND GENE EXPRESSION IN VICIA FABA L. USING LEAF POWDER OF AZADIRACHTE INDICA A. JUSS

A.S. Shehab; S.A.F. Tawab and M.M. Morci

Department of Botany College for Girls, Ain Shams University, Cairo, Egypt.

Experiments were carried out to scrutinize   stimulation effect in the mitotic process of Vicia faba L.when treated with A. indica leaf powder. Remarkably, there was a progressive increase in mitotic indices following treatments. Different applications of neem powder were made on seeds before germination and found to induce an increase in gene expression measured as total soluble protein. Nucleic acids in root tips were also found to increase by increasing the concentrations. Proline content was noticed to increase by increasing concentrations and time of exposure. Biomass productivity was increased following all treatments. Types of abnormalities revealed the induction of spindle disturbance, stickiness, laggards, fragments and macronuclei which lead to the loss of genetic materials. Protein profile revealed the disappearance of some protein bands indicating the genotoxicity of A. indica leaf powder on the biological system under test.

 


34/17 APPLICATION OF INTERSPECIFIC PROTOPLAST FUSION FOR CONSTRUCTION OF NEW HYBRID PROBIOTIC BACTERIA, WHICH INHIBIT SOME OF MYCOTOXIGENIC FUNGI GROWTH IN YOGHURT

A.A. Khattab; K.M. Soliman*; Y.H. Abu Sree*and A.M. Abdel-Salam**

Genetic and Cytology Department, National Research Centre, Giza, Egypt.

*Food Toxins and contaminants Department, National Research Centre, Giza, Egypt.

**Dairy Science Department, National Research Centre, Giza, Egypt.

       The technique of protoplast fusion is potentially an important tool for the genetic manipulation of lactic acid bacteria and hence strains improvement. The inter-specific protoplast fusion between Lactobacillus casei subsp. rhamnosus C34 mutant (P1) and Lactobacillus reuteri (P2) to construct new fusants that could grow in the presence of mycotoxigenic fungi in specific medium and yoghurt were done. Results showed that parents (P1 & P2) and fusants (F1 & F3) inhibited the growth of Aspergillus flavus, A. ochraceus and Fusarium moniliforme. The results indicated that F1 and F3 were highly effective; the complete inhibition (100%) of F. moniliformi was detected in the presence of F3. For A. flavus, P1, P2, F1 and F3 were inhibited the growth by 65%, 73%, 96% and 93.1%, respectively. Also, A. ochraceus was inhibited by 75.9%, 76.0%, 95.8% and 93.1% respectively. Approximately, 81.5%, 81.1%, 96.4% and 97.7% of the total aflatoxins (AFs) were eliminated in by 1012 cfu of P1, P2, F1 and F3, respectively. The most efficient removal of AFs was achieved using fusants. The removal of AFs was rapid process (48h) and binding was dependent on bacterial concentration. The results can be exploited in developing a new approach for detoxification.

 

 

35/17 SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM LEAF CULTURE OF CHRYSANTHEMUM

E.A. Ewais and M.M. Mansour

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

Nasr City, Cairo, Egypt.

High frequency callus induction was obtained from young leaf explants of chrysanthemums (Dendranthema grandiflora) on Murashige and Skoog (MS) medium supplemented with 2,4-D (1.0mg/liter) and kinetin (2.0mg/liter). The callus was unorganized in the beginning, but becamehighly organized and nodular within two months. On MS medium with both auxin (indoleacetic acid, IAA; naphthalenacetic acid, NAA or 2,4-dichlorophenoxyacetic acid, 2,4-D) and cytokinin (kinetin or benzyladenine, BA), plantlets occurred at high frequency through somatic embryogenesis. Mass spectrum was consulted to confirm the metabolites production in both shoot developing in vitro and shoot developing in vivo.   Growth regulators influence to the frequency of callus induction, embryonic callus production and regeneration of plantlets were observed.

 

 


36/17 VARIATION IN STRUCTURE AND METABOLISM OF REGENERATED AND NON-REGENERATED CALLUS OF TOMATO (LYCOPERSICON ESCULENTUM L.)

E.A. Ewais

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

Nasr City, Cairo, Egypt

Initiation of callus was induced from leaf, stem and root of tomato (Lycopersicom esculentum L.) explants on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin. Calluses so formed were subcultured on regenerated media to study morphogenetic capacity of tissues. Meristemoid cells appeared beginning around 2nd week of subculturing. These meristemoids became distinguishable shoot primordial around the 5th week of subculturing. The morphogenesis efficiency was combined with the increases in fresh and dry weights of tissues. Protein, RNA and amino acids contents were stimulated in shoot-forming tissue compared with non-shoot-forming tissue prior and during organogenesis.

 

37/17 PRODUCTION AND CHARACTERIZATION OF A BIOSURFACTANT FROM BACTERIAL SPECIES ISOLATED FROM SOIL

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

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

Structural and biochemical characterization of a biosurfactant produced by 14 bacterial species isolated from soil was performed. The isolates were selected by measuring the oil-film collapsing activity. Preliminary structural determination of extracts by TLC showed that the produced biosurfactant is identical to surfactin. Also, 1H nuclear magnetic resonance and FTIR analysis confirmed that the produced biosurfactant is identical to surfactin. The surfactant yielded a minimal aqueous surface tension values from 27.3 - 36.2 mN/m. The critical micelle dilution of the surfactant were between 16 - 26 fold. The results indicated that most of the isolates have high emulsification activity and are capable of emulsifying hydrocarbons effectively.   The surfactants were measured in culture supernatants by a quantitative method based on its ability to cause blood hemolysis. A high yield of surfactin was obtained from a culture of 3 isolates ranging between 2000 - 6900 mg / l.

 

 

38/17 BIODEGRADATION OF MORPHOLINE AND HETEROCYCLIC ANALOGOUS BY PSEUDOMONAS PUTIDA GR12.2.

M.M. Aly

Biology Department, Faculty of Education, Kafr El-Sheikh, Tanta University, Egypt.

 

Eight different bacteria were screened for morpholine degradation in two mineral media. Pseudomanas putida and Mycobacterium were the most active in morpholine degradation. Pseudomonas putida was selected for more studies. Maximum degradation was achieved at 25 ºC, pH 7.0, shaking at 200 rpm for ten days. Addition of plant growth regulators or different vitamins enhanced the bacterial growth but decreased morpholine degradation. As morpholine concentration decreased, ammonia accumulated and this led to the decrease in the rate of degradation. The growth of Pseudomonas putida increased by increasing morpholine concentration up to 5 g/l, then the growth decreased and no degradation was observed after 17 days using 7g/l. At this concentration, the respiration of Pseudomonas putida decreased and % of cell viability decreased to about 50% (toxic effect). Immobilizatin on activated carbon C40/4 enhanced morpholine degradation. Pseudomonas putida degraded the intermediate compounds   (glycolic , glyceric and glyoxylic acids) through the glycolate route in the presence of 0.1 g/l morpholine as inducer. Ps. putida used also some heterocyclic analogous compounds, which contained only one nitogen atom like piperidine and pyrrolidine in addition to thiomorpholine.

 

39/17 INTERACTIVE EFFECT OF HEAVY METALS AND GIBBERELIC ACID ON MITOTIC ACTIVITY AND SOME METABOLIC CHANGES OF VICIA FABA L. PLANTS

M.M. Mansour

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

The influence of gibberellic acid, two heavy metals (cadmium and lead) and their interactions on mitotic activity and biochemical constituents of Vicia faba yielded seeds was studied. The results showed that different concentrations of cadmium (Cd) and lead (Pb) induced marked inhibition in the contents of soluble carbohydrate, proteins and ribonucleic acid. Heavy metals also induced marked increases of the total percentage of abnormalities. Moreover, different types of chromosomal aberrations were observed in response to heavy metals. On the contrary, gibberellic acid (GA3) stimulated mitotic activity and biochemical constituents of the yielded seeds of Vicia faba. Gibberellic acid could be also counteracted the adverse effects of heavy metals on mitotic index and some metabolic mechanisms of broad bean plants. In addition, electrophoretic profiles of broad bean seeds proteins showed quantitative differences between major bands obtained in control and treated plants.

 

 

40/17 ARCHITECTURE AND BIOACTIVITY OF DEMINERALIZED FREEZE-DRIED BOVINE ALLOGRAFT (DFDBA)

G.T. El-Bassyouni and W.I. Abdel-Fattah

Ceramic Department, National Research Center (NRC), Cairo, Egypt

The present study is concerned with the preparation and structural characterization of demineralized freeze-dried bovine allograft (DFDBA) having two particle sizes [Medium (250-500mm) and coarse (600-700 mm)] compared to underminralized bone fine particle sized allograft (75-120 mm) as a control BA(F). Chemical, spectral and thermal analyses were used for characterizing the three prepared samples. The texture was assessed using SEM/EDXA techniques. The bioactivity of various solids was followed up through measuring the levels of calcium and phosphorus in simulated body fluid (SBF) pre-and post-immersed samples up to 72 hours. Hence, the deposition of newly formed apatitic layer was discussed especially on comparing FT-IR data of pre-and post-immersed solid samples. Results proved that the particle size tremendously affects the efficiency of demineralization process. The coarser the particles, the lower surface area and consequently lower reaction rates. The ionic activity products of (Ca2+) and (Pi) and their corresponding Ca2+/Pi ionic ratio in SBF proved that the particles of medium size exposed its surface for carbonated phosphate deposition more than the coarser ones. However, it is recommended to use DFDBA of optimal size which would have a major role in its osteoinductive capability since, it is more effective than finer and coarser ones.

 

41/17 BIOLOGICAL STUDIES ON POLYMORPHIC CHARACTERS OF CASTOR BEANS “RICINUS COMMUNIS L.”

M.S.A. Soliman and S.F. Badr*

Botany Dept., Fac. of Science, Helwan University, Cairo, Egypt and

*Botany Dept., Fac. of Science, Tanta University, Tanta, Egypt.

Castor bean “Ricinus communis L” (Euphorbiaceae) has a wide ecological and geographical amplitudes. It has different ecological races or ecotypes, which are adapted widely to different environments. Nine fruit samples representing the different populations growing in Egypt were collected in addition to one from both Saudi Arabia and Sudan. Seeds from each sample were subjected to SDS-PAGE protein and RAPD-DNA Gel-Electrophoresis analysis. In the same time, germinated seeds were subjected for chromosome number determination. Results indicated that all the recognised ecotypes of R. communis L. have 2n=20 chromosomes, polymorphic characters on morphological characters (seed weight and volume) as well as molecular bases of genetic origin. The dendrogram of protien banding analysis showed five population groups for R. communis L.

 

42/17 ASCORBIC ACID STABILIZES OXYGEN EVOLVING MACHINERY AND ENHANCES ANTHOCYANINS ACCUMULATION IN TWO CARROT CULTIVARS UNDER SALINITY STRESS

D.Z. Abdel-Kader and A.A.H. Saleh

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

Yellow and red carrot seeds were presoaked over night in 0.0, 0.05 and 0.1M ascorbic acid (ASA). 0.1, 0.2M shock and gradual treatment with NaCl were begun when the seedlings reached 10-cm in length (4-weeks). The different salinity treatments induced significant decreases in O2 evolving rate of the isolated chloroplast. They also significantly decreased total carbohydrates, ALA, Chl.a&b and carotenoids contents. Moreover, salinity treatments induced significant decreases in ascorbic acid content and ASPX activity as antioxidants. It was also found that salinity treatments significantly increased proline content, anthocyanins concentration and electrolytes leakage in both carrot cultivars. Pretreatment with the two ascorbic acid concentrations (0.05 or 0.1M) ameliorate the adverse effect of salinity treatments by stabilizing the oxygen evolving machinery of the isolated chloroplast, increasing proline concentrations and decreasing electrolytes leakage. It also triggers the antioxidant machinery by increasing ascorbic acid content and anthocyanin concentration as well as ASPX activity. Moreover, it was found that yellow and red carrots almost have similar responses to salinity.

 

 


43/17 ANATOMICAL AND PHYSIOLOGICAL RESPONSES OF AVICENNIA MARINA PLANTS TO CRUDE OIL POLLUTION

M.A. Naim

Botany Department, Faculty of Girls, Ain Shams University

Crude petroleum oil has harmful effects on the biological activity of any ecosystem. Five sites of mangrove ecosystems at Al-Qusseir region were surveyed for detecting the presence of petroleum oil pollution. It is found that the five sites are suffering from oil pollution especially the southern end of each site. Crude petroleum oil forms a solid continuous layer which districts the spatial distribution of Avicennia marina propagules as it prevents the penetration of roots. The unsolid petroleum crude oil mixed with the soil didn’t show significant effect on the anatomical and physiological features of Avicennia marina seedlings. It has a remarkable effect on the established trees. The crude petroleum oil slightly alters the structure of leaves and stems but markedly increased the number of lenticels/cm2. Also the pollution caused a disturbance in the activity of the phellogen of the cork layers of the subterranean pneumatophores root parts and vascular cambium of the vascular cylinder of the cable roots. The pollution leads to leaf cytoplasm disruption. Chlorophyll content, soluble carbohydrates, proline content and mineral ion content increased in plant leaves and pneumatophores as affected by crude petroleum oil pollution. Crude oil pollution altered the protein pattern of the leaves. Avicennia marina plants can survive under crude oil pollution but the pollution reduces the plant dispersal.

 

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