Vol. 12, July, 2002

1/12 MICROBIAL DECONTAMINATION OF BIO-ACTIVE GLASS-STUDIES ON THE BIO-ACTIVITY AFTER EXPOSURE TO GAMMA IRRADIATION

O.S. Desouky*;   H.A. Farrag   and   A. El-Ghannam**

Drug Radiation Research Department, Lab. of microbiology,   Physics Radiation Department*, National Center for Radiation Research and Technology, Nasr City, Cairo, Egypt; National Research Center**, Egypt

Microbial contamination to bioactive materials causing biomaterial-centered infection is a problem that limits the extensive use of many bioactive materials in clinical application. In an aqueous environment, ions are released from a bioglass and the pH rises in its vicinity which may influence both growth and colonization of microorganisms. The objective of this study is, to examine the effect of gamma radiation on the bioactivity of the bioactive glass (BG), in order to show if gamma radiation can be used in sterilization of BG without changing the surface properties and bioactivity of it. The bioactivity was examined by studying the formation of calcium phosphate rich layer on the surface of BG. This was carried out by using FTIR and SEM-EDAX. The analysis of the surface showed that the Ca/P rich layer was formed after immersion on the simulated body fluid. The bioactive glass showed no effect of gamma radiation up to 100 Gy on the bioactivity properties. For microbial decontamination of bioglass by gamma radiation, samples of the tested materials BG in the form of granules or discs were tested. The samples were collected from freshly prepared and opened container. In all, 45 microorganisms were isolated from about 5 g of the tested samples. They were belonging to 2 genera and identified as Staphylococcus epidermidis and   Bacillus species. The BG samples were exposed to gamma radiation dose levels from 10 to 130 Gy. The microbial counts of survivors were decreased with increasing the irradiation dose. At a dose level of 100 Gy, three strains were isolated from a total microbial population of around 45 bacterial isolates. An absorbed dose of 130 Gy eliminate all types of microorganisms that contaminate the surface of the tested samples. Antimicrobial activity and growth inhibition of BG on five clinical bacterial and yeast fungal isolates was studied using broth medium and agar plate test. The tested bioglass did not show any clear growth inhibition or antimicrobial action on the tested pathogenic clinical strains. The alkaline pH was considered as inhibitory pH for the isolated microorganisms and all clinical strains used in this study. An alkaline pH a long with gamma radiation dose level of 100 Gy proposed as being sufficient to eliminate all types of microorganisms that contaminate the BG with no effect on the bioactivity properties.

viagra 100 mg durée 2/12 SOME FACTORS AFFECTING ANTIMICROBIAL SUBSTANCE PRODUCED BY THE BLUE GREEN ALGA NOSTOC MUSCORUM.

F.T. Zaki and A.A. Fathi

Botany Department, Faculty of Science, Cairo University, Egypt and Botany Department, Faculty of Science, El-Minia University, Egypt.

The heterocystous nitrogen fixing Cyanobacterium prix viagra lyon Nostoc muscorum was isolated from the soil, purified and grown at 30C in the modified Watanabe medium under white flourscent light. les dosages du viagra Nostoc muscorum secretes antibiotic, which is biologically active against other strains of cyanobacteria and bacteria. The data obtained here showed that the antibiotic produced by kamagra ne marche pas Nostoc muscorum was found to be very active against other prokaryotic organisms either cyanobacteria or various strains of bacteria and thus is suggested to be a broad spectrum antibiotic. It also inhibited the growth of le viagra en tunisie Chroococcus sp. and reduced population density and chl-a content at different concentrations, being lethal at higher doses. The antibiotic treatment increased the protinease activity. The antibiotic produced by generique4u cialis Nostoc muscorum was maximized under the following conditions: BG-11 +N medium, 330 mol m-2s-1 light intensity, 1.5 gl-1 nitrate-N concentration, and phosphate concentration of 0.6 gl-1.

3/12 PURIFICATION AND CHARACTERIZATION OF MILK CLOTTING ENZYMES, FROM PHASEOLUS VULGARIS AND PISUM SATIVUM FRUITS

S.T. El-Sayed

Biochemistry Department, National Research Center,

Milk clotting enzymes (M1 and M2) were extracted from fresh fruits of Phaseolus vulgaris and Pisum sativum, respectively. They were highly purified by precipitation with ammonium sulfate (30-70% saturation) and acetone (30-50%) followed by gel filtration on Sephadex G-120 column and two successive Sephadex G-200 columns. The enzymes M1 and M2 were purified 15.06 and 20.67 times with yield recovery of 20 and 40.1 %, respectively as compared to the crude preparation. The homogeneity of the purified enzymes was proved by polyacrylamide gel electrophoresis patterns. A single band was observed for each sample. The molecular weights estimated by gel filtration on Sephadex G-200 were 66 and 60 KD for M1 and M2, respectively. The optimum enzyme activity of M1 and M2 was at pH 4 - 4.6 and 45ºC, with an activation energy of 25.38 and 31.56 Kcal/mole, respectively. Their activities were considerably stable up to 40ºC for 60 min with only 33% loss of its activity and marked loss upon heating at 70ºC. The two enzymes M1 and M2 are glycoprotein in nature. They gave ultraviolet spectrum characteristic of protein. Calcium chloride stimulate (accelerate) milk clotting activity up to 0.11%, whereas sodium chloride was inhibitor of the activity even at the low concentrations. The milk clotting activities of M1 and M2 enzymes were activated by treatment with 5.0 mM Fe+3, Mg+2 and EDTA by 12 to 49% of its activity. They are able to produce white soft curd with good quality. They had remarkably high milk clotting activities with fairly weak proteolytic activities. The enzyme M1 possessed higher milk clotting activity per proteolytic activity ratio than that of M2.

4/12 IMPROVEMENT OF L-LYSINE PRODUCTION BY GAMMA IRRADIATED REGULATORY MUTANTS OF CORYNEBACTERIUM GLUTAMICUM

A.I. El-Batal

National Center for Radiation Research and Technology (NCRRT),

Nasr City, Cairo, Egypt.

The production of L-lysine by fermentation was studied, using homoserine deficient (Hse-) and gamma irradiated S-(2-aminoethyl) L-cysteine-resistant (AECr) regulatory mutants of Corynebacterium glutamicum in shake flasks. A study was performed on the effects of different carbon sources, mineral nitrogen and some metabolic activators (biotin and some amino acids) as constituents of nutrient media on the production of L-lysine by a culture of C. glutamicum strain (Hse-). The most favorable components for the biosynthesis of the product were selected, and the optimal concentrations of these components were determined. Gamma irradiated regulatory mutants resistant to the L-lysine analog (AECr) have been isolated from homoserine auxotroph (Hse-) parental strain in order to obtain mutants desensitized to the feedback inhibition. Most of the (AECr) resistant mutants enhanced isolates have improved production of L-lysine. The superior enhanced mutant isolate No. γ7-AECr 34 is exhibiting the highest L-lysine production (30.5 gL-1) with relative increase (158.1%) in production yield than parental strain. Stimulations in L-lysine production were observed when low concentrations of permeability enhancers Tween (20, 40, 60 and 80) as well as dimethylsulfoxide (DMSO) were added during the course of fermentation, however Triton X-100 had a negative effect. Tween 60 (0.1%, v/v) addition gave the highest level of lysine yield (36.8 gL-1).

5/12 XYLITOL PRODUCTION FROM CORN COBS HEMICELLULOSIC HYDROLYSATE BY CANDIDA TROPICALIS IMMOBILIZED CELLS IN HYDROGEL COPOLYMER CARRIER

A.I. El-Batal and S.A. Khalaf*

National Center for Radiation Research and Technology (NCRRT),
P. O. Box 29, Nasr City, Cairo, Egypt.

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

         The ability of five yeast strains (locally isolated) to ferment xylose to xylitol were screened using a corn cobs hydrolysate. viagra pour femme c est quoi Candida tropicalis proved to be the best producer. The effects of culture conditions, namely initial pH, nitrogen source and yeast extract concentration, on xylitol production were evaluated. The conditions for batch production of xylitol, using kamagra oral jelly siparis C. tropicalis immobilized growing yeast cells in hydrogel copolymer carrier composed of polyvinyl alcohol and hydroxyethyl methacrylate (PVA/HEMA) produced by radiation polymerization, have been optimized. Strategies for enhancing bioconversion of hydrolysate by immobilized cells, including the influence of the most important operational variables (age of inoculum, initial concentrations of immobilized cells and oxygen limitation) were discussed. The effects of supplementation of fermentation medium with different concentrations of metabolic inhibitors (monofluoro- acetic acid or monochloroacetic acid) on xylitol production by immobilized cells were studied. The highest level of xylitol (41.0 gL-1) was obtained in fermentation medium supplemented with 10 mg % of monofluoroacetic acid. Different concentrations of methanol as an activity modifier supplem- ented to fermentation medium were screened for their capacity to improve xylitol production by immobilized cells. The maximum xylitol production (45.2 gL-1) was reached at 1.5 ml % of methanol addition. In repeated batch fermentation (4 days for each run), the immobilized cells retained their ability to produce the superior xylitol yield (48.5 gL-1) in presence of 10 mg % of monofluoroacetic acid and 1.5 ml % methanol supplemented together to fermentation medium. The results also demonstrated that during about 5 weeks of repeated batch mode under optimized conditions, the final xylitol concentrations decreased gradually in nine cosecutive runs.

6/12 PRODUCTION, CHARACTERIZATIONS AND APPLICATION OF GEOTRICHUM CANDIDUM LIPASE IMMOBILIZEDIN HYDROGEL COPOLYMER CARRIER

A.I. El-Batal and K.S. Atia*

National center for Radiation Research and Technology, P.O. Box 29, Nasr City, Cairo, Egypt

*Nuclear Research Center, Radioisotop Applications Division, Abou-Zaable,

Cairo, Egypt

Production of extracellular lipase by Geotrichum candidum was substantially enhanced when 1.0% (v/v) olive oil was supplemented to basal fermentation medium. Lipase activity as high as 48.7 Uml-1 of culture medium was obtained at optimum conditions after 4 days of incubation. A partially purified preparation of lipase shows specific activity of 20.1 Umg-1 protein. The prepared enzyme was immobilized by entrapment onto polyvinyl alcohol/ polyacrylamide (PVA/ PAAm) copolymer hydrogel carrier produced by gamma radiation polymerization. The specific activity of the immobilized lipase was significantly influenced by the amount of enzyme loaded into the copolymer hydrogel carrier. The optimal value of immobilized protein was 2 mg/ ml gel matrix blend. The effect of temperature, pH, operational stability and kinetic studies indicated that the immobilized lipase showed a higher stability than the free one. A conversion of 82 and 83% was obtained upon esterification of oleic or palmitic acid with methanol.

achat viagra en algerie 7/12 PRODUCTION OF PECTINASES BY GAMMA IRRADIATED INTERSPECIFIC HYBRIDS OF ASPERGILLUS SP. USING AGRO-INDUSTRIAL WASTES.

A.I. El-Batal and M.A. Khalaf

National Center for Radiation Research and Technology (NCRRT) P.O. Box 29, Nasr City, Cairo, Egypt.

Protoplast release in pectinolytic mutants strains of effect viagra et alcool Aspergillus niger UV5 Ade- and A. terreus UV2 Cys- was described. Also, interspecific protoplast fusion between the above mutant isolates was done to breed new strain producing pectinases from lemon peel. Further, upgrading of this production was enhanced through radiation treatment. Optimum yield of protoplast UV5 and UV2 was obtained after 4 h when crude lytic enzymes of Trichoderma harzianum were added in 0.05 M citrate-phosphate buffer pH 5.0 containing 0.7 M KCl. Isolated protoplasts in UV5 and UV2 were capable of a moderate regeneration frequency 54% and 36% when 0.7 M KCl was used as osmotic stabilizer. Polygalacturonase (PG) and pectinlyase (PL) production were not modified during the process of regeneration. Protoplast fusion between UV5 Ade- and UV2 Cys- by polyethylene glycol treatment produced prototrophic fusants with a fusion frequency of 2.5-4.4 x 10-4. The reversion frequency of the protoplasts was low (5.5-8.2 x 10-5). One prototrophic hydrid (HG) produced higher amount of PG (11.52 Um-1) and PL (6.80 Uml-1) when grown on medium containing 1% lemon peel for 3 days. A high yield of PG (38.4 Uml-1) and PL (22.5 Uml-1) production was recorded by HG gamma irradiated active isolate (HGg2 when grown on 3% lemon peel for 5 days.


8/12 PROPERTIES OF FIBRINASE FROM PENICILLI

H.M. El-Shora, and S.A. Khalaf*

Botany Department, Faculty of Science, Mansoura University, Mansoura

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

            The highest fibrinase production by P. digitatum; P. aculeatum and P. cyclopium was observed on the fourth day of growth. Fibrinase was purified with specific activities of 410, 370 and 188 U mg-1 protein, respectively. The optimal pH values were 8.0, 7.0 and 7.5, respectively. Maximum activity was observed after 60 min for the first organism and 80 min for both the second and the third ones. The optimal temperatures were 45, 40 and 50 0C, respectively. Zn2+, Co2+ and Cu2+ cations had an inhibitory effect. Monovalent cations K+1 and Na+1 had no significant effect on fibrinase activity from any tested organism. The chelating agents: ethyleneglycoltetraacetic acid (EGTA) and ethylelnediaminetetraacetic acid (EDTA) inhibited fibrinase activities. The SH-reagents 5,5`-dithiobis-(2-nitrobenzoic acid) (DTNB), phenylmercuricacetate (PMA) and mersalylinhibited fibrinase with PMAbeing the most potent inhibitor. Na azide, Na malonate and Na fluoride were good inhibitors. Phenylmethyl-sulphonyl fluoride (PMSF) inhibited fibrinase activities and revealed the necessity of serine in enzyme catalysis. The inhibitory effect was most pronounced on the P. aculeatum fibrinase. Gibberellic acid (GA3) increased fibrinase activities from the three penicilli when added to the culture medium.

9/12 THE ALLELOPATHIC EFFECT OF ZYGOPHYLLUM ALBUM ON MORPHOLOGY, ANATOMY AND PHOTOSYNTHETIC PIGMENTS OF VIGNA MEMBRANACEA

M.M. Migahid and M.M. Elkhazan

Biological and Geological Department, Faculty of Education, University of Alexandria, Alexandria, Egypt and Botany Department, Faculty of Education for Girl’s Jeddah, Kingdom of Saudi Arabia

The effect of Zygophyllum album as an aqueous fresh or dry extract and shoot residue (dry shoot powder) on Vigna membranacea was studied. The morphological and anatomical structures of V. membranacea hypocotyl were changed with the application of Z. album extract. Fresh or dry shoot extract inhibited germination and growth of V. membranacea seedlings. This inhibition was much more pronounced by using the extract of fresh plants. Dry powder of Z. album applied on surface or incorporated in the soil has reduced growth and chlorophyll content of V. membranacea, but increased osmotic pressure. The surface applied powder has more effect on V. membranacea than incorporated one, especially on the root growth. Root system of V. membranacea seemed to be more sensitive to the allelopathic effect of Z. album than shoot system. The environmental stress as salinity increased the allelopathic effects of Z. album.

10/12 RAPD-PCR ANALYSIS OF SOME FUNGI ISOLATED ON BLACK LIQUOR

M.M. Amer

Botany Department, Faculty of Scince Benha University

Genotypic differences and relationships among eleven fungal isolates (Aspergillus flavus, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Aspergillus clavatus, Pencillium notatum, Paecilomyces divaricata, Paecilomyces silvatica, Fusarium oxysporium, Fusarium moniliferum and Fusarium poae) isolated from province El-Qalubia were studied based on DNA polymorphism. Seven random 10-mer primers were used in the RAPD-PCR analysis, but only two primers (OPX-03 and OPV-19) were able to generate repeatable informative products among the DNA samples of the studied genotypes. A maximum of 42 DNA bands were detected as PCR products for the two primers, 36 of them were polymorphic. A total of 14 polymorphic bands were scored as unique ones. Combination of all data provides a considerable potentiality for discriminating each fungal isolate by one or more unique bands. RAPD data were combined together and used to construct a dendrogram that estimates the relationships among the studied Aspergillus isolates. Based on this dendrogram the studied isolates were separated into two main groups. The first group includes the four isolates Aspergillus flavus, Aspergillus terreus, Aspergillus fumigatus and Aspergillus clavatus while the second one comprises the isolate Aspergillus niger. The similarity coefficients among the three Fusarium isolates scored from polymorphism across RAPD analysis were used to identify their genetic relationships.

 

11/12 BIOREMEDIATION FOR THE DECONTAMINATION OF SEWAGE AND INDUSTRIAL WASTEWATER BY FERN AZOLLA CAROLINIANA.

M.A.Hemada and E.M. Mostafa, Botany Department

Faculty of Science, Alexandria University, Egypt.

The growth of Azolla caroliniana grown on different sources of sewage water was significantly reduced particularly on human and industrial sewage. This reduction was accompanied with a marked decrease in macronutrients accumulation and an increase of heavy metals concentration. Irrigation of broad bean plants with different Azolla-treated sewage water resulted in a significant increase in growth parameters, total carbohydrate and total protein contents comparing to Azolla-untreated sewage water. The increase of macronutrients concentration in the shoots and roots of broad bean plants grown in various sewage water was accompanied with a decrease in heavy metals concentration.  

12/12 PULLULAN PRODUCTION BY AUREOBASIDIUM PULLULANS USING BEET MOLASSES

S.A. Miligy and H. Abdel Kareem

National Center for Radiation Research and Technology, Department of Microbiology, Nasr city-Cairo- Egypt

The production of pullulan by Aureobasidium pullulans ATCC 42023 from beet molasses in shake flask culture was investigated. The effect of molasses concentration, inoculum level, pH, added nitrogen sources and gamma irradiation were studied to maximize pullulan production. At pH 6, 10% was the optimum molasses concentration and inoculum volume where pullulan production reached 14.02 gL-1. Pullulan production   increased reaching 16.82 gL-1 at 0.04% ammonium phosphate concentration. At dose level 0.25 kGy of gamma irradiation, the highest achieved amount of pullulan was   19.82 gL-1 comparing with 18.48 gL-1 of control one (basal medium).

13/12 EXPRESSION OF THE WILD TYPE P73 GENE IN LUNG CANCER TISSUES

M.A. Mohamed, N.M.A.Fahim, L.A. Helalah*and S.A. Affifi**

Departments of Microbiology and Immunology, Chest* and Internal Medicine**, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

P73, a new p53 homologue, has been identified.It is unclear whether this protein functions as a tumor suppressor gene.To clarify whether p73 might be involved in lung carcinogenesis,p73 mRNA expression was examined in lung cancer tissue biopsies in 17 patients with different types of lung cancer as well as in corresponding normal lung tissues using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), with β-actin as a control. P73 expression in lung cancer tissues, using P1 and P2 primers, was significantly higher than in normal lung tissues (2.11 ± 0.69 vs 0.023 ± 0.015, P < 0.001).Using another set of primers, P3 and P4, confirmed the results of PCR using P1 and P2. P73 expression using P3 and P4 primers was also significantly higher in lung tumors than that in normal lung tissues (0.39 ± 0.47 versus 0.023 ± 0.015, P< 0.01). This study suggests that p73 may play an important role in lung tumorigenesis,and suggests the need for further functional analysis of the role of p73 in lung carcinogenesis.

14/12 STUDIES ON SEED COAT AND POLLEN GRAINS MORPHOLOGY OF SOME TAXA OF ZYGOPHYLLUM (ZYGOPHYLLACEAE) AND THEIR TAXONOMIC IMPLICATIONS

A.A. El-Ghamery, M.M. Mansour, E.A. Abdel-Azeem and A.M. Kasem

Botany & Microbiology Dept., Faculty of Science, Al-Azhar Univ., Madinat Naser, Cairo, Egypt.

Seed morphological characters, seed coat sculpture and pollen grain features of nine taxa of Zygophyllum were estimated with light and scanning microscopy in order to assess their taxonomic significance. These taxa are Z. simplex, Z. decumbens var. decumbens, Z. decumbens var. megacarpum, Z. coccineum, Z. album var. album, Z. album var. amblyocarpum, Z. berenicense, Z. aegyptium and Z. propinquum. Seed shape and size as well as cell wall sculpture allow the recognition of two groups among the studied taxa. The variations in seed coat features are helpful in the sectional and intra-group delimitation. The result was in agreement with the previous taxonomic studies based on of the morphological characters. In contrast, the variation in pollen grains is of no significant value at infra-specific level.

15/12 PURIFICATION AND CHARACTERIZATION OF PECTINLYASES FROM ALTERNARIA ALTERNATA AND TRICHODERMA LONGIBRACHIATUM

A.A. Khalil

Department of Biological Science, Faculty of Education, Ain Shams University, Heliopolis, Roxy, Cairo, Egypt.

Two pectinlyases [EC. 4.2.2.3] were isolated from the cell free filtrates of Alternaria alternata (Fr.) Keisser (PLI) and Trichoderma longibrachiatum Rifai (PL II)- after solid state fermentation using orange peel- by ammonium sulfate precipitation, gel filtration through Sephadex G-100 and ion exchange chromatography on DEAE-Cellulose. The two enzymes were purified ~30 and 46-folds with total recovery of 56 and 60%, respectively. Maximal activities of the two enzymes were recorded at pH 4.8 and 45°C for PLI while pH 5.2 and 50°C were the optimal for PL II. The two enzymes were found to have a good thermostability. Km values were calculated to be 0.21 and 0.35 mM, respectively.

16/12 FINGERPRINT OF SOME BACTERIA ISOLATED FROM SOME WASTE PRODUCTS BASED ON DNA POLYMORPHISM

S.S. Abd El-Salam

Botany Department, Faculty of Scince Benha University

Genotypic differences and relationships among six Acetobacter and two Gluconobacter isolates were studied based on DNA polymorphism. The survey conducted on the randomly amplified polymorphic DNA based polymerase chain reaction (RAPD-PCR) analysis showed considerable differences among the studied isolates. Nine random 10-mer primers were screened. But only two primers (OPX-03 and OPV-19) were able to generate repeatable informative products among the DNA samples of the studied isolates. A total of 51 polymorphic bands were generated by the two primers. A total of 19 polymorphic bands were scored as unique ones. The primer OPX-03 was found to be the more potent in generating unique bands than OPV-19. The former primer generated 11 unique bands while the latter primer produced 8 unique bands. Combination of all data provided a considerable potentiality for discrimination betweenisolates either by one or more unique bands or a group of combined class pattern. Similarity coefficients were calculated based on all pooled data. A dendrogram that estimates the relationships among the studied Acetobacter genotypes was constructed. Based on this dendrogram the studied isolates were separated into 2 main groups. The first group comprises A. rancens, A. oxydans and A. roseum while the second one includes the three isolates A. aceti, A. xylinum and A. suboxydans. The highest similarity value (0.692) was recorded between the two isolates A. rancens and A. oxydans while, the lowest similarity (0.00) was recorded between the two isolates A. xylinum and A. oxydans.

 

17/12 EFFECT OF DIFFERENT TEMPERATURES AND PRESENCE OR ABSENCE OF SEED COAT ON GROWTH, METABOLISM AND PROTEIN BANDING PATTERNS OF LUPINUS TERMIS SEEDLINGS

A.A.A. El–Mashad, A.M.A. Mohammed and E.A. Kamel

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

       In the present study, the effect of different temperatures on growth, endogenous growth regulating substances, total carbohydrates, total protein, activities of certain related enzymes and protein pattern on coated and uncoated seeds of Lupinus termis cv. Giza (1) are presented. The results indicated that the radicle and plumule lengths, number of lateral roots, fresh and dry weights were increased in both coated and uncoated seeds of Lupinus termis during germination when the temperature was raised from 20 to 25oC. On the other hand, at 30 oC all those parameters were decreased. The magnitude of growth in uncoated seeds is much more pronounced than that in coated ones. Total carbohydrate contents, endogenous growth regulating substances, protease and catalase activities were increased from 20 to 25oC and decreased at 30oC, while total protein contents, growth inhibitors, peroxidase and IAA-oxidase activities recorded reverse patterns. The protein banding patterns of Lupinus termis seedlings extracted in Tris-HCl, showed the presence of at least 28 different bands, having approximate molecular weights ranging from 13.44 to 96.09 KD. Considerable variations in total number and M. Wts. of protein bands between coated and uncoated germinated seeds for one or six days were recorded.

 

 

18/12 BIOLOGICAL EFFECTS OF GAMMA IRRDIATION ON THE FLESH FLY, PARASARCOPHAGA SURCOUFI VILL.

(DIPTERA: SARCOPHAGIDAE)

G. M. Aly

National Center for Radiation Research and Technology (NCCRT) P.O. Box 29

Nasr City,Cairo - Egypt

Exposure of the full grown pupae (10-day-old) of the flesh fly Parasarcophaga surcoufi Vill to different dose levels of gamma irradiation had a depressing effect on the percentage of adult emergence, longevity and the reproductive potential (the mean number of deposited larvae per female). It was found that the lower dose levels of gamma radiation have a great effect on the reproductive potential. The two dose levels of 16 and 12 Gy were considered as the optimum sterilizing doses which induced complete sterility in the adult male and female respectively. That is probably due to the fact that the oviviviparous flesh flies are more radiosensitive to gamma radiation than the oviparous ones.   The sex ratio fluctuated at the lower doses, whereas at the higher doses the male ratio increased over female ratio. In addition the adult longevity decreased insignificantly towards the sterilizing dose, whereas the adult female longevity was not affected by irradiation. The pupal lethal dose was found to be 120 Gy.

19/12 COMBINATION OF GAMMA IRRADIATION AND CYPERMETHRIN AS AN INTEGRATED   CONTROL MEASURE FOR THE FLESH FLY PARASARCOPHAGA DUX THOMSON (DIPTERA: SARCOPHAGIDAE)

G.M. Aly   and F.E. Megahed

National Center for Radiation Research and Technology (NCCRT) P.O. Box 29,

Nasr City, Cairo-Egypt.

The determined LD10 of the synthetic pyrethroid cypermethrin against Parasarcophaga dux (2.80 µg/ larva) was topically applied on the tergum of the 3-day-old, third instar larvae. This dose had no significant effect on the adult emergence and sex ratio. However the reproductive potential as well as the male longevity were significantly reduced by applying the sterilizing dose level (13 Gy), the substerilizing dose levels (10, 11 and 12 Gy) either separately, or in combination with the cypermethrin insecticide. The gamma - irradiated males either separately or in combination with cypermethrin - treatment   were sexually competitive with the normal males at ratios of 15:1:1 and 20:1:1 (irradiated males : normal males : normal females respectively) with competitiveness values of 0.65. and 0.93 respectively. This was also held true for the radiosterilized males which can compete effectively with (cypermethrin-treated males + normal males, in   equal numbers )   at   a ratio of   10:1:1 (irradiated males : cypermethrin-treated males +   normal males: normal females respectively) with a competitiveness value of 0.83.

20/12 TAXONOMIC IMPORTANCE OF SEED CHARACTERISTICS OF SOME SPECIES OF CASSIA AND SENNA (SUBFAMILY: CAESALPINIOIDEAE – LEGUMINOSAE)

H.A. Hussein, S.F. Khalifa*, A. Ghareeb and N. Fawzi**

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

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

**Central Laboratories Unit, U.A.E. University, United Arab Emirates.

Seed characteristics of 12 species representing; Cassia and Senna, were examined with scanning electron microscope (SEM) in the light of their taxonomic importance. The characters proved to be taxonomically useful include: variations in shape, colour, areole, lens, hilum and micropyle. In addition, the surface scan peculiarities, as viewed under SEM, either at a low magnification up to 150x or at a high magnification between 1500-2500x, afford a valuable taxonomic evidence reliable for delimiting the species investigated at both the generic and specific levels.

21/12 PRODUCTION OF PROTEIN AND FIBER FROM BEET WASTES

M.A. Osman, A.M.S. Galal, A.G. Allam and H.E.M. Abu El- Eneen,

Dept. of Food SCI. and Tech., Fac. of Agric., Al-Azhar Univ.

Sugar beet leaves and pressed pulp were used in this study which consider a by-product of sugar beet industry. Fat extraction of the raw leaves and pulp by hexane (1:5w/v) gave defatted samples had high contents of protein, fiber, ash and carbohydrate except fat content was low compared with the raw samples. Extraction of different proteins from defatted leaves by different methods: Chloroplastic protein prepared by 0.1N NaOH method (1:20 W/V) at pH6, from the same solvent prepared of cytoplasmic protein at PH 4. While crystal protein prepared by 0.5% sodium metabisulfite method (1:20 W/V) at pH 4 and at 4 C°. Crystal protein had high values of protein (65. 62%), digestibility (33.5-62.0%), foam capacity (130), foam stability (125), emuslsion (52), water absorption (440) and oil absorption (418), but the values of fat (0.47%), fiber (3.5%), ash (4.0%), carbohydrate (26.41%), poly pHenols(0.12%), oxalate (o.24%),   pHytic acid (0.20%) and chloro pHyll (0.015%) were low followed by cytoplasmic and chloroplastic proteins respectinely. Fiber extraction from defatted pulp by soaking in NaOH solution at   pH 10-12 for 1-2 hours at 50-100 C°, washed, soaking in 1 NHCI solution, washed, dried and ground. The fiber extracted had high content of fiber (43.52%) and low contents of protein (4.21%), fat (0.96%) ash (6.27%) and carbohydrate (43.04%) compared with defatted pulp. Analysis of fortificated balady bread with different fiber levels chemically indicated that the contents of moisture, protein, fat and carbohydrate were decreased and the values of ash and fiber were increased by increasing of fiber levels. Also, analysis of frankfurter chemically appeared that the values of protein, ash and carbohydrate were increased, but the values of moisture and fat were decreased by increasing of added leaves proteins. Quality properties in the bread and frankfurter were organoleptically evaluated. Generally, it could be concluded that the balady bread containing 5% of fiber had acceptable quality. Also, frankfurter containing 3% of different leaves proteins had high acceptability and quality.

22/12 TAXONOMIC CRITERIA OF THE CHARACTERISTIC FEATURES IN SEEDS OF SOME SELECTED SPECIES OF CAESALPINIOIDEAE-LEGUMINOSAE

H.A. Hussein, S.F. Khalifa*, A. Ghareeb and N. Fawzi**

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

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

**Central Laboratories Unit, U.A.E. University, United Arab Emirates.

       Seeds of 17 species, representing 14 genera of Caesalpinioideae, were examined under the scanning electron microscope (SEM) to define the diversity of attributes of micropyle, hilum, lens and sculpturing patterns. The differences in the characters of the micropyle, hilum and lens seem to be distinctive for seeds of some of the species investigated. The testa surface exhibited six basic sculpturing patterns namely; levigate, rugulate, reticulate, foveolate, papillose-rugose and ill-defined. Furthermore, intergraded forms within the basic patterns were, also, encountered. The distribution of such patterns presented an indispensable criterion for delimitation, primarily, at the species level and, sometimes, at the generic rank. Additionally, the relevance of the testa sculpturing patterns with Bentham’s classification (1865) was discussed.

23/12 MOLECULAR CHARACTERIZATION OF SOME SELECTED LOCAL PEACH (Prunus persica L.) CULTIVARS IN DAKHALIA GOVERNORATE

H.Z. Hassan, N.M. Mansour* and E.A. Abd El-Hady**

Dept. Bot., Fac. Sci., Ain Shams University, Cairo, Egypt

*Hort. Res. Inst., Agric. Res. Cent., Mininstry of Agriculture, Cairo, Egypt

**Dept. Genetic and Cytology, National Research Center, Dokki, Giza, Egypt.

Eleven peach (Prunus persica L.) selections representing six local cultivars growing at different locations at Dakhalia Governorate were fingerprinted by SDS-protein and random amplified polymorphic DNA (RAPD) markers. The cultivars are Sultani, Hegazy, Shamy, Fark, Mawy and Neely. Low polymorphism level was scored in the protein profile of the studied peach selections. However, the resulted protein profile was considered as a general fingerprint of these peach selections. On the other hand, RAPD results revealed a high level of polymorphism among the studied genotypes. Of thirty-one 10-mer random primers screened, ten gave reproducible polymorphic DNA amplification patterns. The number of the generated bands was primer and genotype dependant. A total of 180 amplified DNA bands were generated across the studied genotypes with 147 bands being polymorphic. Of these bands, 30 unique ones were detected. All primers except OPB-07 gave a number of unique bands that were used for the discrimination of peach selections. No primer was able to generate a unique banding pattern for all of the eleven peach selections. However, combinations of all data derived from the SDS-protein banding pattern and the ten random primers produced unique fingerprint for each selection. Similarity coefficients among the studied selections were estimated after pooling all protein and RAPD data. The obtained dendrogram divided the studied genotypes into two subgroups and shows that peach selections that belong to a particular cultivar were grouped together in a single branch.

24/12 CLONING SECONDARY EMBRYOS FROM MICROSPORE-DERIVED EMBRYOS OF CANOLA: INDUCTION AND ELECTRON MICROSCOPY INVESTIGATIONS

H. Emara*, S. Abd Al-Tawab** and M. Zaki

Plant Biotechnology Lab., Institute for Efficient Productivity, University of Zagazig, Zagazig, Egypt

*Institute of Genetic Engineering and Biotechnology, University of Minufiya,                   Sadat City, Egypt

**Botany Department, Faculty for Girls, University of Ain Shams, Cairo, Egypt

Initiation of successful microspore culture was marked by an increase in microspore volume, which occurred during the first 12 hours in culture medium. Within 48 hours in culture, first cell division was observed in the microspore suspensions. The division was unsynchronized but symmetric and resulted in formation of two identical cells enclosed within the pollen exine. Continuous and rapid division of the formed cells resulted in formation of mature pollen-derived embryos in culture medium. When the formed bipolar cotyledonary embryos were isolated and cultured on medium supplemented with the cytokinin thidiazuron, secondary embryogenesis commenced. Embryos that cultured on medium devoid of thidiazuron showed little response to elements in culture medium. During early stage in culture, only a few secondary embryos were initiated and the number increased till the fourth week in culture, after which there was no further increase in number of secondary embryos. Secondary embryos appeared first as globular structures and became visible by the second week in culture. Secondary embryos are not initiated synchronously but various stages of development were observed emerging on pollen-derived embryos.   Addition of the cytokinin thidiazuron to the culture medium enhanced secondary embryogenesis. The higher level of the cytokinin thidiazuron was incorporated into culture medium, the more secondary embryos were initiated and developed. Application of thidiazuron at 50µM in culture medium resulted in stimulation of embryogenesis with maximum number of secondary embryos developing at the fourth week in culture. Monthly subculture of secondary embryos on fresh medium supplemented with thidiazuron resulted in maintenance of the secondary embryonic potential for long period of time. Ultrastructural survey indicated that cells of secondary embryos featured dramatic increase in amount of lipid material that found spread throughout the cytoplasm. On the other hand, cytoplasm of cells that form the parental tissues were devoid of lipid. Moreover, electron microscopy study confirmed the presence of large starch grains in plastids of parental tissues but not in cells of secondary embryos.   Differences between types of organic materials that present in the cytoplasm of cells forming haploid and secondary embryos have been observed.   The recovery of numerous numbers of secondary embryos on pollen-derived embryo is considered a unique and unequivocal system for large-scale production of synthetic seeds. Moreover, production of secondary embryos is a unique method for cloning numerous embryos from single genetically engineered microspore.

25/12 CONSERVATION AND REGENERATION OF THE ENDANGERED ACACIA NILOTICA SUBSP. TOMENTOSA (BENTH.) BRENAN (ACACIA ARABICA (LAM.) WILLD.) THROUGH TISSUE CULTURE TECHNIQUES

A.M. Shaheen and M.H. Omar

Aswan Faculty of Science, Botany Department, South Valley University

Tissue culture of Acacia nilotica subsp. tomentosa (A. arabica) was achieved using different parts of seedlings (shoot tip, cotyledonary leaves and hypocotyl) explants on Murrashige and Skoog (MS) medium supplemented with different hormones, namely 6-benzyladenine (BA) and combination of cytokinins and naphthalene acetic acid (NAA). The maximum survival rate of shoot tip explants were observed with BA (2 mg/l) or BA:NAA (0.5:2 mg/l); callusing was only observed when shoot tip explants were cultured on BA (4 mg/l) or BA:NAA (0.5:2 mg/l). The maximum shoot regeneration from shoot tip was obtained with BA (4 mg /l) with 10 shoots per explant. The highest survival rate of cotyledonary leaf explants were observed with BA (2 and 4 mg/l) and BA:NAA (0.5:2 and 1:0.5 mg/l). The maximum callusing rate from the hypocotyl (10%) was obtained with BA:NAA (0.5:2 mg/l). It was revealed that shoots can be regenerated from seedlings of Acacia nilotica subsp. tomentosa (A. arabica), either directly by culturing terminal buds on BA:NAA (0.5:2 mg/l) or indirectly from callus, which also developed from terminal bud, and from cotyledonary leaves (BA:NAA; 0.5:2 and 1:0.5 mg/l) and from callus developed from hypocotyl by culturing on BA:NAA (0.5:2 mg/l).

26/12 IMPROVEMENT OF OXYTETRACYCLINE PRODUCTION BY IMMOBILIZED STREPTOMYCES RIMOSUS IN k-CARRAGEENAN IN BATCH AND REPEATED BATCH CULTURES.

H.A. El-Enshasy

Bioprocess Development Dept., Genetic Engineering and Biotechnology Research Inst., Mubarak City for Scientific Research, Alexandria, Egypt.

Immobilization of Streptomyces rimosus was carried out by k-carrageenan and k-carrageenan modified gel beads for production of oxytetracycline. The maximal antibiotic production of 250 mg/l was obtained when using immobilized spores in a concentration of 1´107 spores/ml. This value was about 175 % higher compared to the corresponding free cell culture. To enhance the gel stability and minimize the cell escapement from the gel beads, the immobilized beads were treated with polyethyleneimine (PEI) as hardening agent. The treated beads showed a higher production capacity with minimal escapement compared to the untreated beads. Further improvement of the production process was achieved by pre-cultivation of immobilized beads in rich vegetative medium to achieve a higher cell density inside the immobilized beads prior cultivation in the production medium. Repeated batch production of oxytetracycline by immobilized cells in k-carrageenan gel was achieved for 8 repeated batches without a significant decrease in the antibiotic production. After the first batch phase of 96 h, the reduction of batch time from 96 h to only 48 h does not decrease oxytetracyline production by immobilized cells.

27/12 APPLICATION OF YEAST DEBAROMYCES HANSENII AS A BIOCONTROL AGENT AGAINST POSTHARVEST BLUE AND GREEN MOLD OF LEMON FRUITS CAUSED BY

PENICILLIUM SPP

A.A. Hussain

Botany Department, Women,s College, Ain Shams University, Cairo, Egypt.

The osmotolerant yeast strain D. hansenii ( Y-1268) was evaluated for its activity in reducing postharvest decay on lemon fruits caused by two species of Penicillium (blue and green mold) isolated from the surface of infected fruit . The tested yeast had a significant   reduction potential ( 72.3 and 80.5% ) against the radial growth of the two Penicillium, sp respectively on a synthetic medium at 285C compared with the other used media . Image Analysis indicated, no destruction of the fungal hyphae and no direct interaction between the yeast cells and pathogens, while chlamydospores were appeared as a resistance against the antagonistic stress. Also D. hansenii did not produce antibiotic substances in the culture. Furthermore, no reduction of fungal growth was exhibited by each of autoclaved yeast cells, cell free filtrate and the enriched media when grown at 285C on a synthetic medium, these in contrast with both a viable yeast cells and precultured medium. Efficacy of the antagonist was affected by the concentration of both the yeast cells and fungal spores, it was most effective at 109C FU / ml of the yeast cells. Because of the tested yeast was halotolerant , where it can tolerate NaCl at 8.0%, some inorganic salts such as Ca Cl2 , Ca CO3 , KCl and NaCl were tested to evaluate their effect on the yeast efficacy against Penicillium decay. Calcium chloride at 1.5% was significantly the most effective salt applied with yeast cells in wound treatment that enhance its efficacy against the molds. It reduced the infection rate into 1.3 and 1.7 % with green and blue mold respectively. Whereas, salt solutions without yeast cells applied either on wounds or in potato dextrose medium did not reduce the fungal growth . Moreover, the results indicated that competition for nutrients may play an important role in the biocontrol on lemon fruit and/or in synthetic medium, the biological control decreases when nutrients are amended. The percent infection on artificial wounds of fruits was reduced to 12.7 % when treated with washed viable yeast cells against green mold, whereas it increased to 89.3 % with blue mold when the wound site amended with nutrients. There is no significant difference between the two species of Penicillium under all conditions.

28/12 LOW FAT-HIGH NUTRITIVE BUTTER
USING CHICK PEA (CICER ARIETINUM)

Z.A.M. El-Atawy

Animal Production Research Institute, Agriculture Research Centre

Low fat butter was obtained from the conventional butter (control) by replacement ratios with chick pea paste (CPP) 10, 20, 30 and 40% (w/w). The product was stored at 5oC for 15 days. The chemical analysis, keeping quality and organoleptic properties of the product were carried out. The caloric values and cost of the ingredients were calculated. Results showed higher moisture, protein, ash and mineral contents, titratable acidity percentages, peroxide value (PV) and thiobarbituric acid, while lower fat percentages, caloric value and cost ingredients of the resultant butter were obtained with increasing the replacement ratio of CPP. Slight decreases were gradually noticed in organoleptic scores between treatments. Generally, an acceptable low fat butter could be successfully made by mixing CPP with butter at a level up to 40%.   The product had high spreadability, good body and texture that remained stable up to the end of storage period with favourable flavour, high nutritive value and lower cost.

29/12 EFFECT OF SOME RADIOMODIFIERS AND GAMMA IRRADIATION ON THE FERTILITY AND COMPETITIVENESS OF THE MOSQUITO CULEX PIPIENS COMPLEX L.

G.M. Aly

National Center for Radiation Research and Technology (NCRRT) P.O. Box 29, Nasr City, Cairo-Egypt

The effective dose (LC50/24h) of two radiomodifier solutions namely caffeine and cysteine, was determined by applying concentrations of 0.75% w/v and 0.90% w/v respectively to 0-h pupae of the mosquito Culex pipiens Complex L. The least rate of mortality was recorded after 15   and 10 minutes (optimum exposure time) for caffeine and cysteine respectively. Caffeine solution acted as a radiosensitizer where the adult emergence was reduced from 73.33% in case of applying gamma radiation alone to 58.33% , 69.66% and 51.31%   in case of pretreatment with caffeine solutions after latent periods of 30, 60 and 90 minutes respectively with highly significant difference at 30 and 90 minutes. On the other hand, cysteine solution acted as a radioprotector where some recovery of adult emergence was achieved as the adult emergence was increased at the same latent periods to reach 75.00%, 78.33% and 74.66% respectively with insignificant difference. The egg hatchability showed a minor recovery when the irradiated males were pretreated with the effective dose of cysteine solution as it increased from 0.00 to 3.19 % and 4..21% after latent periods of 30 and 60 minutes respectively. The competitiveness value of the irradiated males at the ratio of 15 : 1 : 1 irradiated males : normal males : normal females was slightly increased from 0.65 to 0.77 when these males were pretreated with the effective dose of the radioprotector cysteine solution. Whereas, The competitiveness value at the same ratio was decreased from 0.65 to 0.37 when the irradiated males were pretreated with the effective dose of the radiosensitizer caffeine solution.

30/12 THE INTERACTION OF SALINITY AND IAA ON GROWTH AND SYNTHESIS OF SOME IMPORTANT METABOLITES IN DUNALIELLA SALINA (TOED.)

M.M. Allam and M.A. Hemada

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

Growth and some metabolic activities of Dunaliella salina (Toed) grown in the presence of different grades of salinity with and without exogenously added IAA were monitored. The results revealed that IAA treatments induced a stimulatory effect for all growth parameters, chlorophylls, protein and glycerol contents of D. salina to a degree higher than the control itself. The degree of stimulation depended on the level of salinization and concentration of the exogenous additives of IAA.

31/12 MOLECULAR CHARACTERIZATION OF SALMONELLA PARATYPHI B GENE (RECA) WHICH CONFERS FLUOROQUINOLONE RESISTANCE IN ESCHERICHIA COLI DH5Α

M.A.M. Yassien, H. Ewis*, C. Lu* and A.T. Abdelal*

Department of Microbiology and Immunology, Faculty of Pharmacy, Ain-Shams University, Cairo, Egypt.

*Department of Biology, Faculty of Arts and Science, Georgia State University, Atlanta, Georgia , USA.

A genomic library from a strain of Salmonella paratyphi B that exhibits multidrug resistance was constructed in Escherichia coli DH5a and two recombinant plasmids were obtained. One of these plasmids, pNFXR5, conferred selective fluoroquinolones phenotypic resistance in E. coli. Sequence and subcloning analysis showed that it is the presence of RecA which confers resistance to fluoroquinolones. The results of protein analysis of crude extract and crude outer membrane proteins of the obtained fluoroquinolones resistant E. coli clones showed an overexpression of 38 KDa protein as compared to the wild type E. coli DH5a. Western blot analysis with anti-recA confirmed that the 38 KDa protein is RecA protein. The overexpression of RecA may increase the level of resistance of E. coli to fluoroquinolones through the coprotease activity of RecA to trigger the cascade of the SOS regulatory system. These results suggests that overexpression of RecA causing high level of selective fluoroquinolones resistance in E. coli.

32/12 NATURAL CONTAMINATION OF EGYPTIAN WHEAT WITH MOLDS AND FUMONISIN B1 TOXIN AND ITS METABOLIC TOXICITY DURING GRAIN GERMINATION .

M.A. Foaad And A.M.A. Mohamed

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

Natural contamination of Egyptian wheat with molds and fumonisin B1 toxin had been detected. Thirty-two samples from different goverenorates and six cultivars were subjected to isolation and identification of fungal mycoflora to the genus .The fungal species isolated were Alternaria spp, Aspergillus spp, Aspergillus flavus, Fusarium spp,Curvularia spp, Penicillium spp and Rhizopus spp. Thirteen different strains of Fusarium spp were isolated from surface and sixteen from non surface sterilized grains, the contamination of wheat grains with FB1 detected by TLC reconfirmed quantitatively using HPLC technique. The existence of fumonisin B1 in tested samples were within the range 8.14 to 54.30 µg/g wheat grains. To appraise the FB1 potential in wheat grains with special reference to the growth of producing Fusaria isolates; the most sensitive samples to FB1 producing by different isolates were inoculated with isolates and growth was determined by glucosamine mg/g inoculated grains minus the control uninoculated grains. The results are also showed that FB1 started to be detected after four days of inoculation and no toxin was detected before. The results also showed that FB1 is phytotoxic on auxins, gibberellins, metabolism of sugars, amylases, protease and protein which significantly caused some morphological changes in root and shoot system during the first six days of germination. The electrophoretic profile of different protein bands of germinated wheat grains treated with fumonisin B1 mycotoxin was also investigated which had drastically affected by fumoinisin B1.
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