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Vol. 11, May, 2005

1/11 NESTED-PCR TECHNIQUE FOR THE DETECTION OF THE HERBICIDE-RESISTANCE (BAR) GENE IN THE GENOME OF SOME STREPTOMYCES SPECIES

H.I. Abdel-Fattah

Department of Agricultural Microbiology, Faculty of Agriculture, Zagazig University, Zagazig, Egypt

The herbicide resistance (bar) gene encodes a phosphinothricin-acetyltransferase (PAT) is widely used in transgenic plant production to induce herbicide resistant plants. Therefore, in this work, the presence of such gene was detected in the DNA genome of eight local Streptomyces strains. To reach such goal, nested-polymerase chain reaction (PCR) technology using two internal primers flanking a PCR product with a size of 450 bp was used. Prior to that the ability of the applied Streptomyces strains to grow on the recommended doses (RD) or 2 or 3 folds of Sencor and BASTA herbicides were determined. The growth of Streptomyces strains withstanding the toxicity of the herbicide was determined and recorded as no growth (-), weak (+), moderate (++) and abundant (+++). The experimental results showed that Sencor at 3RD inhibited the growth of S. lincolnensis ST55, S. venezuelae ST71 and S. umbrinus ST86, while the remainder 5 strains showed weak growth (S. rectiviolaceus and S. roseolus) to moderate growth (S. albosporeus subsp labilomycaticus, S. herbaricolor and S. aureomonopodiales). No abundant growth at 2 either or 3 RD of Sencor was found.In case of BASTA, results showed that the tested streptomycete strains were varied in their possibility to grow on 4 and 6 g/L BASTA when added to the growth medium (starch nitrate agar medium). Again, S. albosporeus subsp labilomycaticus, S. herbaricolor and S. aureomonopodiales appeared abunant growth in case of all concentrations of BASTA, while S. lincolnensis STd55 appeared moderate growth and both of S. venezuelae ST71 and S. umbrinus ST86 appeared weak growth when BASTA herbicide was added with a concentration of 3 folds of the RD. As an interesting result, the amplified PCR products confirmed this physiological result. Whereas, the Nested-PCR products were amplified only from the DNA of the genome of S. albosporeus subsp labilomycaticus, S. herbaricolor, S. aureomonopodiales, S. lincolnensis ST55, and S. venezuelae ST71. On the other hand, no PCR products were amplified from S. rectiviolaceus, S. roseolus and S. umbrinus ST86 strains as well as the negative control.

 

 

2/11 MINERALIZATION OF FORMULATED ALACHLOR LASSO® AS CARBON AND NITROGEN SOURCES BY CUNNIGHAMELLA ELEGANS LENDNER ADAPTED TO ALACHLOR

M.F. Abdel Latteff, M.B. Mahmoud, H.M.S. Khalifa and S.I. Ibrahim

Plant Protection Department, Faculty of Agriculture (Cairo), Al-Azhar University.

The aim of this study was herbicide to use Cunnighamella elegans Lendner to biodegrade the formulated herbicide alachlor (Lasso®). Two isolates were used, the parent isolate (PI) and adapted isolate (AI). The adapted one which obtained by several exposure of the parent isolates to formulated alachlor. The results indicated that the PI isolate grow on complete medium amended with different concentrations of alachlor up to 300 ug/ml being 81% reduction of the colony growth, however this isolate did not grow on medium containing 400ųg/ml. Interestingly, adaptation process, about 10 exposures, produced an isolate (AI) could grow on medium containing 700 ųg/ml alachlor. The results also indicated that both isolates grow on carbon-or nitrogen – free medium amended with alachlor, however, the AI grew more effectively than the PI, especially on nitrogen free medium. Additionally, total carbohydrates and proteins determination in mycelial matrix of both isolates confirmed this investigation, whereas total carbohydrates and proteins were higher in AI than PI after culturing on liquid media amended with alachlor. Using a bioassay technique, the EC50 values, concentration of alachlor as ųg/g peat moss which inhibit the growth shoot and root systems of millet by 50 % , were determined . These values were determined after 0.0, 10, 20 and 30 days of stored peat moss supplemented with alachlor alone or mixed with the fungal isolates. Based on these values both the fungal isolates reduced the herbicidal effect of alachlor to millet growth and the AI of the fungus reduced this effect more effectively and more rapidly than PI C. elegans. It could be mentioned that the AI isolate of C. elegans could be used for disposing the excess stock of formulated alachlor.

 

 

3/11 THE RHIZOSPHERE MICROORGANISMS AS ANTIFUNGAL PRODUCERS: A COMPARATIVE STUDY

A.A. El-Mehalawy

Microbiology Department, Faculty Of science, Ain Shams University, Cairo, Egypt.

This investigation was aiming to compare between the different groups of rhizosphere microorganisms (actinomycetes, mold fungi, yeast fungi and bacteria) according to their effect on the plant pathogenic fungi through the production of antifungal compounds. It was reported that all the tested species have the antagonistic activity against the tested pathogenic fungus Rhizoctonia solani. The yeast and mold fungi are characterized by their antagonistic activity which is higher than that of actinomycetes and bacteria. It was indicated that each of Gliocladium virens and Pseudomonas flourescens produce 6 active compounds of different Rf values followed by Trichoderma viride, Streptomyces lydicus, Saccharomyces unispora, Candida incommunis and Arthrobacter ramosus each produce 5 active compounds, while S. graminofaciens produce only 4 active compounds. It was noticed also that the active compounds of Rf values; 0.13, 0.3and 0.5 were common in the culture filtrates of one or two species of three groups of the four tested ones.

4/11 POTENTIALITY OF ACTINOMYCETE ISOLATES AGAINST DERMATOPHYTIC FUNGI

1- BIOLOGICAL, TAXONOMICAL STUDIES AND ENVIRONMENTAL FACTORS AFFECTING THE ANTIFUNGAL PRODUCTION OF

STREPTOMYCES KANAMYCETICUS EHE-68

M.H. Yassin; M.F. Ghaly*; S.M. Shash and E.H. El-Doraidy

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

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

Three hundred actinomycetes isolates were isolated from different soil samples collected from cultivated soil at Qaluobia, Egypt. Out of 300 actinomycetes isolates, 13 ones were able to exhibit the antifungal activity against the tested dermatophyte fungi, namely Tricophytone rubrum, Tricophytone mentagrophytes and Micosporium canis. The isolate No. 68 was found to be the best one of them, so it was identified up to species level, as follows: Streptomyces kanamyceticus EHE-68. Streptomyces kanamyceticus EHE-68 was grown on starch nitrate broth either at static or shaked conditions (180 rpm). It was found that the maximum values of antifungal activity were at shaked condition. The maximum antifungal activity of S. kanamyceticus. of extracellular protein and growth rate after 10 days of incubation. The experimental organism was able to tolerate a wide range of temperatures (20-50°C). The optimum temperature for maximum antifungal activity, extracellular protein and growth rate was 30°C. S. kanamyceticus. was able to grow on a pH range of 4-10. The optimum pH value for antifungal activity, extracellular protein and growth rate was 8.0.

 

5/11 PRODUCTION, PURIFICATION AND CHARACTERIZATION OF PROTEASE PRODUCED BY BACILLUS MYCOIDES ON ARTICHOKE WASTE

M.E. Osman, N.M. Sidkey*, H.H. El-Hendawy and M.M. Attwa

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

*Botany and Microbiology Department, Faculty of Science (Girls), Al-Azhar University, Egypt.

Wastes of three plants (peas, artichoke and spinach) and water hyacinth were collected from Montana factory, Kalyobeya Governorate and River Nile, Sharkeya Governorate, Egypt. These wastes were air dried, ground and used as a natural medium for bacterial growth. Thirty two bacterial isolates were isolated from the collected plant wastes and screened for production of cellulases, pectinases and proteases, the highest level of these enzymes were produced by three isolates symbolized as : Pe4 (from peas); Ar10 and Ar8 (from artichoke), respectively. These isolates were identified as Bacillus polymyxa (Pe4), B. mycoides (Ar8) and B. subtilis (Ar10). Studying the optimum conditions for enzyme production in media containing the plant wastes revealed that B. mycoides Ar 8 was the most potent isolate in protease production. The enzyme was purified by (NH4)2SO4 precipitation, ion exchange and gel filtration column chromatographies. The purified protease was found to be homogenous by SDS-PAGE with molecular weight of 55 kDa. The enzyme was optimally active at pH 7.4 and temperature of 35oC.

6/11 ANTIFUNGAL EFFICACY OF – IRRADIATED AND NON-IRRADIATED ESSENTIAL OILS AGAINST SOME PATHOGENIC AND NON-PATHOGENIC SOIL FUNGI

S.A. El-Aasar and E.M.M. Atia

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

The information concerning with antimicrobial activity and the action mechanism of plant extracts have led to study the ability of essential oils as potential antimicrobial agents. Essential oils of chamomile, eucalyptus, fennel, geranium, lemon, peppermint and neem plants greatly reduced the total count of fungi and bacteria inhabiting soil and stored seed belonging to family solanaceae (tomato, aubergine, okra, pepper and potato tubers). Geranium, fennel and eucalyptus oils showed more markedly inhibitory effect than other tested oils either by direct contact or fumigation. The combined effect of oils concentration and gamma-irradiation was found to be a limiting factor on bacterial and fungal count .Where the gamma- irradiation dose was increased from 10 to 40 KGy and coupled by increasing the oil concentration from 500 to 4000 ppm, the inhibitory effect of total microbial count increased. Moreover, the antimicrobial activities against the tested microorganisms was also studied, Alternaria alternata growth was completely inhibited when treated with eucalyptus oil at 1000 ppm irradiated with 40 KGy, limon with 4000 ppm (non-irradiated), fennel with 1000 ppm, chamomile with 4000 ppm and geranium with 2000 ppm. While Alternaria lycopersici growth was completely inhibited when treated with fennel oil at 2000 ppm. However the same fungal species showed more resistance to oils of eucalyptus, chamomile and geranium, but a complete inhibition of the fungal growth was recorded when treated with 4000 ppm. Aspergillus flavus, A.niger and Trichoderma viride were more sensitive and completely inhibited with low concentration of eucalyptus, lemon, chamomile and geranium oils. The capability of Fusarium oxysporium for inducing wilt disease in tomato plant was also reduced to 55 – 65 % inhibition, when the infested tomato seeds were soaked with water extract of fennel seeds for 2 or 5 hours, respectively. While 3000 ppm essential oil of fennel gamma-irradiated at 30 KGy caused 85 and 90 % inhibition after 2 and 5 hours of soaking, respectivily. Essential oils of eucalyptus and geranium at 3000 ppm irradiated at 30 KGy with gamma rays had 70 and 85 % inhibition effect on disease occurring of seeds infested with F. oxysporium when compared with control.

 

7/11 DNA FINGERPRINT OF DENTOPHILIC CANDIDA ALBICANS

M.H. Eid* and N.E. Yousef

*Pedodontic Department, Faculty of Oral and Dental Medicine, Cairo University

Department of Microbiology, Faculty of Pharmacy,Zagazig University

A group of 25 Candida albicans isolates from patients with oral candidosis, were tested for chondroitin sulphatase and protease activity. Of the 25 tested isolates, C19 was protease non-producer and chondroitin sulphatase producer (Pr - and Ch+) and C21 was protease producer and chondroitin sulphatase non-producer (Pr and Ch-) and the others were chondroitin sulphatase and protease producers (Pr + and Ch+). The most virulent Candida albicans isolates from patients with candidosis were chondroitin sulphatase and protease producers (Pr+ and Ch+). Using scanning electron microscope, it was found that isolated Candida albicans strains did not maintain their yeast morphology but gave rise to blastospores, hyphal filaments and germ tubes covered the root canal walls after 5 days of incubation. After 10 and 15 days of incubation, budding yeast cells were attached tightly to dentinal walls and hyphae penetrated to dentinal tubules. DNA fingerprinting of Candida albicans strains of different isolated strains and different morphology (blastospores, hyphae, budding or germ tubes) proved to have the same DNA bands pattern of fingerprint. The results suggest that protease and chondroitin sulphatase production, hyphae and germ tube formation play an important role in adherence of Candida albicans to dentin and virulence of Candida albicans.


8/11 IMPACT OF SOME PHENOLIC COMPOUNDS (AS POLLUTANTS) ON GROWTH AND SOME METABOLIC ACTIVITIES OF ALGAL SPECIES INHABITING DIFFERENT WATER HABITATS AT EL-FAYOUM GOVERNORATE, EGYPT.

M.H.M. Abdel-Rahman, M.S. Abdel-Kareem*, S.H. Shaalan*, A. Khaleafa*

and M.Y.A. Gad

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

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

The effects of three well-known phenolic compounds on growth and some metabolic activities of three dominant algae inhabiting different water habitats in El-Fayoum Governorate (Egypt) were evaluated. The results showed that phenolic compounds inhibited algal growth. Meanwhile, the sensitivity of the tested algae towards phenolic compounds differed according to the algal species, the type and concentration of the phenolic compounds, and the duration of culturing. Also, the results emphasized that low concentrations of some phenolic compounds promoted the growth of some organisms, while the toxicity of the tested phenolic compounds at high concentrations increased, and caused reduction in growth of all the tested algae. Concerning the metabolic activities, the results revealed that, with some exceptions, the biosynthesis of pigments, proteins, polysaccharides and total carbohydrates by the three algal species were inhibited, while the biosynthesis of soluble sugars and amylases activities were enhanced under phenolic stress. Also, the results emphasized that the higher the toxicity and concentration of the phenolic compounds, the more inhibition of the lipase activity and the more accumulation of lipids. Different patterns of the synthesis of amino acid fractions in the three studied species were observed in accordance to the effect of phenolic compounds.

 

9/11 NUTRITIVE VALUE IMPROVMENT OF SUGAR BEET PULP THROUGH BIOLOGICAL TREATMENT.

1-SELECTION AND OPIMIZATION OF CULTURE CONDITION FOR SELECTED FUNGAL STRAIN PERFORMANCE

M. Fadel, S.M. Allam; T. M. Al-Bedawi, H.H. El-Amari and S.H. Mohamed

Microbial Chemistry Department, National Research center, Giza , Egypt.

Animal production Department, Faculty of Agriculture, Cairo University.

Animal production Research institute, Agricultural Research Center, Ministry of Agriculture, Egypt.

Four fungal strains namely Trichoderma viride F-.416, T.harzanum F.415, T.harzianium F.115 and Aspergillus niger F- 909 were evaluated to select the most efficient one for subsequent biological treatment of sugar beet pulp (SBP) under solid state fermentation. The obtained results revealed that T. viride F.416 was more efficient than other tested strains for increasing crude protein (CP) content as well as decreasing hemicellulose cellulose and pectin Both IVDMD and IVOMD reached its maximum in SBP treated by T .viride F-416. Cultural condition including initial pH value, temperature, moisture content and incubation period as well as mixed culture with yeast were studied for optimizing. Promising results were obtained at initial pH value 5.5, temperature 33oC,.solid: moisture ratio 1:2 ratio and Saccharomyces cerevisiae F-25 addition at 0.1%(w\w). Under above conditions protein content in SBP increased from 7.9 to 19.4%. crude fiber was decreased from26.98 to 15.07% Reduction Cellulose, hemicelluloses and pectin were recorded. IVDMD reached its maximum to 66.72% at 0.1% level instead of.50.80% in untreated substrate.

10/11 NODULATION AND NITROGEN FIXATION BY LUPINUS VARIUS L. A WILD TYPE OF LUPINE GROWN UNDER DIFFERENT WATER REGIMES IN SOIL FROM SOUTHERN EASTERN DESERT OF EGYPT.

M. Younis

Botany Department, Faculty of Science, South Valley University, Aswan, Egypt

The effect of water stress on the growth and symbiotic relations of Lupinus varius L. a wild type of Lupine, grown in Wadi Eigat in the southern part of the eastern desert of Egypt was evaluated in pot experiments. Four treatments of water regimes, each was applied to Lupinus varius L. inoculated with mixed inoculum of Bradyrhizobium sp. strains (TAL 1303 and TAL 1407), 20 Days after planting (DAP),for growth and symbiotic relations studies. Wadi Eigat soil embodied a higher proportion of fine and course sand (67.36% and 17.13%, respectively) and accordingly is classified as loamy sand soil. Significant decrease in dry weight of shoots and roots, nodule population and mass, total protein and Leghaemoglobin content of nodule cytosol, specific   nitrogenase activity at low water regimes was recorded. Despite this significant decrease, it appears, however, that plants subjected to severe drought (W3 and W4) exhibit remarkable tolerance indicating that Lupinus varius L. is tolerant to severe water stress in desert soil. This study presents Lupinus varius L. as a good candidate which is water-stress tolerant and of medicinal and economical values and are able to fix appreciable amount of N2 for propagation under arid conditions such as those prevailing in the southern desert of Egypt.

11/11 PRODUCTION AND PARTIAL PURIFICATION OF A LIPASE FROM ASPERGILLUS WENTII

H.A. El-Minofi

Natural & Microbial Products Chemistry Department, National Research Centre, Dokki, Cario, Egypt.

Among 10 fungal strains which fermented at different incubation periods (48, 72 and 120h) on using cottonseed (C.S.) as a cheap nutrient containing lipid residues, Aspergillus wentii was the most potent microorganism to produce lipase (228.6 u/ml) after 72h on using C.S. cake (contains 0.5g% lipid). On studying some parameters in order to obtain maximal lipolytic activity using A. wentii, the most suitable conditions were as follows: The production medium contains (g%): C.S. 8; diamonium hydrogen citrate, 3; KH2PO4, 0.2; KCl, 0.05; MgSO4 7H2O, 0.05 and olive oil, 0.2; the pH was adjusted initially to 6, the medium inoculated by 5% (2.6 x 107 spores of A. wentii) and incubated at 30°C on a shaker incubator (200 r.p.m.) for 3 days (lipolytic activity 363.58 u/ml). Partial purification of this enzyme was achieved by 60% ammonium sulphate saturation, therefore the specific enzyme activity and the purification fold were increased (48.13u/mg protein and 2.08 fold respectively).


12/11 NUTRITIONAL REQUIREMENTS OF A NEW CYCLOSPORIN A PRODUCING ASPERGILLUS TERREUS CULTURE

L.A.R. Sallam, A.H. El-Refai, H.A. El-Minofi, A.A. Hamdy and I.S. Abdel-Salam.

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

A series of experiments were conducted to outline the nutritional requirements of a new cyclosporin A (Cy A) producing fungus; namely Aspergillus terreus. The studies included the role of carbon, nitrogen sources, inorganic salts as well as the amino acid supplements. The promising fungus gave maximal amounts of Cy A on the production medium composed of g/l: glucose, 5 (repeated addition of after 7 days incubation) ; peptone, 1 ; KH2PO4, 0.15 ; KCl, 0.25 ; L-leucine + L-valine, 0.2% of each, pH 6.3. The sterilized medium should be inoculated with 2% of fungus inoculum of 48 hr old, shaked at 200 r.p.m. for 10 days.

13/11 PREVALENCE OF STRONGYLOIDES STERCORALIS AND   OTHER INTESTINAL PARASITES AMONG SCHOOL CHILDREN IN DIFFERENT LOCALITIES IN SHARKIA GOVERNORATE, EGYPT

K.A. Mohamed* and M.A. Abd El-Rehim**

Parasitology Department, Faculty of Medicine (*Damietta and **Assiut Branch),

Al–Azhar University

Strongyloides stercoralis is a parasitic nematode with a worldwide distribution but especially common in the tropical and subtropical areas. In Egypt, recent infection rate is not known as little attention was directed towards this parasite. So, the aim of this work is to identify the prevalence of Strongyloides stercoralis infection among school children in some villages in Sharkia Governorate in relation to clinical presentation and environmental factors. Seven hundred and ninety seven primary and preparatory school children from Sharkia Govemorate were included in this study. There ages ranged between 6-14 years old of both sexes. This study comprised 271 students from El-Khers, 159 from El-Aziziia, 142 from Kafr Shalshalamon, 123 from Meet yazeed and 102 from Menia El-Kamh city. All stool samples were subjected to direct smear examination, formol-ether concentration technique, Petri dish and Harada Mori culture methods. The results showed that the prevalence of S. stercoralis was 0.5% [4 cases out of 797 students examined]. The first one was from Menia El-Kamh [1/102 (0.9%)] and the other three cases were from El-Khers [3/271 (1.1%)]. One case was detected by direct smear while two cases were detected by Harada Mori culture method. The four cases were detected by formol-ether concentration and Petri dish culture methods. Mixed infection of Strongylides, Giardia lamblia, Hymenoleps nana,Entameba histolytica, Enterobius vermicularis, Hymenoleps diminuta, Taenia species and Trichurus trichurus were found in seven cases (0.9%). Three cases in El-Aziziia, two cases in Kafr Shalshalamon, one case in Menia El-Kamh and another one case in Meet Yazeea. It was concluded that the poor environmental factors and bad sanitary conditions in certain areas [e.g.: Kafr Shalshalamon] contributed to the high prevalence of intestinal parasitic infections. This may be attributed to the difficulty in obtaining a clean water source, lack of proper sewage and garbage disposal, together with poor health education. Also, it was concluded that the formol-ether concentration method is superior to the direct smear method in detection of various intestinal parasites [237 (29.7%) versus 198 (24.8%)].

14/11 EVALUATION OF NUCLEIC ACID AMPLIFICATION PROTOCOLS IN DIAGNOSIS OF PULMONARY MYCOBACTERIUM TUBERCULOSIS

M.A. El Demellawy, A. Abdel Wahab, E.M. Emad*, K.M. Kandeel* and

M.K. El Awady**

Medical Biotechnology Department, Genetic Engineering & Biotechnology Research Institute (GEBRI), Mubarak City for Scientific Research & Technology Applications (MuCSAT), New Borg El Arab, Alexandria, Egypt

*Department of Biochemistry, Faculty of Science, Alexandria University, Alexandria, Egypt

**Department of Biomedical Technology, National Research Center (NRC),

Cairo, Egypt

Early detection of Mycobacterium tuberculosis (TB) in clinical samples has become more important in the control of tuberculosis. This study was designed to compare the routine TB diagnosis based on microscopy examination, and culture with 3 nested PCR based assays using as templates either, IS6110, mtp40 or 85B-RNA in the diagnosis and treatment follow up of pulmonary tuberculosis. Sputum specimens from 250 patients clinically diagnosed to have pulmonary tuberculosis were utilized. Group I: 120 patients with suspected TB infection; group II: 70 patients relapsed after treatment, group III: 30 patients not responding to treatment and group IV: 30 patients subjected to follow up every two months during the treatment. In the present study, PCR and RT-PCR were able to detect TB DNA and RNA in sputum from patients with negative smear test as well as samples with negative culture. False positives were observed in DNA PCR at the end of successful treatment. On the contrary, RT-PCR of 85B RNA is more specific and sensitive method for detecting only viable mycobacteria. Our data altogether indicate that RT-PCR is fast, sensitive and specific method for diagnosis and follow-up of TB infection.

15/11 SOME PROPERTIES OF KERATINASE OF BACILLUS PUMILUS FH9 STABILIZED BY COVALENT COUPLING TO SOLUBLE POLYSACCHARIDE

H.A. El-Refaie, M.A. Abd El Naby, M.H.I. El-Araby and A.F. Abdel Fattah

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

Keratinase from Bacillus pumilus FH9 was stabilized (modified) with periodate oxidized pectin. The modified (conjugated) enzyme retained 71.75% of the original specific activity exhibited by the native keratinase. Compared to the native enzyme, the conjugated form exhibited higher optimum pH, optimum reaction temperature, energy of activation, higher Km (Michaelis constant) and improved thermal stability but lower Vmax maximal reaction rate). The thermal deactivation of the native and conjugated enzyme obeyed the first order kinetics. The calculated half-life values of heat of inactivation at 50, 60, 70 and 80° C for the conjugated keartinase were about (623, 452, 188, 143 min), respectively, whereas at these temperatures the native enzyme was less stable (half life of 102, 74, 30, 8 min respectively). The deactivation rate constant at 80°C for the conjugated keratinase was about 2.1 x 10-3 which is lower than that of native enzyme (39x 10-3). The activation energy for denaturation of the native enzyme was about 5.07 Kcal/ mol. The effect of some metal ions on the activity of the conjugated keratinase has also been investigated.

16/11 BACTERIAL CLEAVAGE OF SOME ORGANOPHOSPHORUS PESTICIDES AND MOLECULAR CHARACTERIZATION FOR SOME PHOSPHOESTERASE ISOMERS

Y.A. Mawgoud and F.A. Bughdadi*

Botany Department, Faculty of Science, Cairo University, Giza 12613, Egypt.

*Science Department, Teachers College, Makkah, P.B.2064, Saudi Arabia.

Five bacterial isolates able to use some organophosphorus pesticides (OPs) as sole phosphorus sources were obtained from 12 soils collected from cultivated farms at Kaha, Egypt and Makkah, Saudi Arabia. Non of the studied OPs could serve as a sole carbon source. Isolates were derived from slightly acidic soils (pH 6.7). All isolates utilized diethylphosphate oxon (paraoxon) and diethylphosphate thions (diazinon and parathion). Only two isolates were most versatile to use, in addition, dimethyl thion (methyl parathion). No isolate was capable to biodegrade phosmet (dimethyl phosphate methylenoxy thion). The two versatile isolates were identified as Pseudomonas sp. F1 and Pseudomonas sp. F2. Agarose gel analysis revealed that their biodegradation gene is a chromosomal gene. The native polyacrylamide gel electrophoretogram of their phosphoesterase enzyme revealed 2 isomers for Pseudomonas sp. F1 (M. wt 36.4 & 37.7 K Da) and 4 isomers for Pseudomonas sp. F2 (M. wt 35.6, 36.2, 37.4 & 37.9 K Da).

 

17/11 EFFECT OF PLANTARICIN UGI ON SURVIVAL OF BACILLUS CEREUS ATCC 14579ENDOSPORES IN- VITRO AND IN SOME EGYPTIAN FOODS.

G. Enan and A.H. Saad*

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

*Department of Food Hygiene, Faculty of Veterinary Medicine, Suez Canal University, Ismalia, Egypt.

The effect of the bacteriocin plantaricin UGI on survival of B. cereus ATCC 14579 endospores was studied in brain heart infusion broth (BHI) broth and in some Egyptian food viz. cooked rice; minced meat; tomato paste; pasteurized milk; Kareesh cheese. The spore germination percentages were calculated in both samples and controls. After 48 hrs of storage, the percentage values of spore germination reached their maximum values in controls. They were ranged from 28% to 45% in controls; but reached only 6-9 % in all treated food samples. The percentage values of survived spores were calculated as ranging from 15.4% to 77.7% in all the treated foods samples tested. Consequently the percentage values of in activated spores ranged from 84.6% to 22.3%.By further storage of samples up to 96 hrs, foods samples treated with plantaricin UGI showed marked decline of survived spores. The percentage values of inactivated spores ranged from 78.8% to 100% and the percentage values of survived spores were ranged only from 0% to 21.2%. Almost, the inactivation power of plantaricin UGI against B. cereus endospores was apparently more in liquid foods than in solid foods.


18/11 MOLECULAR AND BIOCHEMICAL STUDIES ON THE ADAPTABILITY OF A SALINE SOIL ISOLATE OF STAPHYLOCOCCUS AUREUS FOR HIGH SALT CONDITIONS

N.H. Abbas, H. El Rabey and M. Osman

Genetic Engineering and Biotechnology Research Institute, Minufiya University, Sadat City, Egypt

A salty soil isolated bacterial strain was identified as Staphylococcus aureus. This isolate has a high molecular weight plasmid and showed resistance to many antibiotics such as streptomycin, penicillin and chloramphinicol. The bacteria can grow at salt concentration up to 3 M. The SDS-PAGE shows the presence of a 220 KDa protein band at the low salt growth conditions. This band disappears when the cell was turned to high salt concentration. The salt tolerance mechanism of the isolated bacteria includes the accumulation of proline amino acid. However, the use of degenerative primer for the conserved ectoine region indicated that the bacteria have no conserved ectoine operon sequence in its genome.  

 

19/11 STIMULATORY EFFECT OF PROPIONIBACTERIUM SHERMANII ON THE GROWTH OF BIFIDOBACTERIUM BIFIDUM

F.A. Fathi

Dairy Department, National Research Centre, Dokki, Cairo, Egypt

The ability of metabolites from Propionibacterium shermanii to simulate the growth of Bifidobacterium bifidum was investigated. Growth of B. bifidum in sterilized skim milk was stimulated by the addition of different concentrations of sterilized metabolites of P. shermanii. B. bifidum attained high cell populations growing in sterilized skim milk supplemented with 2% sterilized metabolites of P. shermanii. The efficiency of P. shermanii metabolites to stimulate the growth of B. bifidum in soft cheese (Domiati cheese) over long storage period (15 days) at 7°C was evaluated. Addition of 2% sterilized metabolites produced by P. shermanii caused stimulation for B. bifidum growth, whereas it caused complete inhibition of yeast and mould. Based on these results it was concluded that stimulation of B. bifidum was due to production of bifidogenic growth factor by P. shermanii. It may be play an important role in the regulation of bifidobacterial populations in dairy products.

 

20/11 BIODEGRADATION OF POLY -Β- HYDROXYBUTYRATE PRODUCED BY AZOTOBACTER CHROOCOCCUM USING STREPTOMYCES VIOLACEUS

S.M.M. El- Sabbagh

Botany Department, Faculty of Science, Minoyfiya University

Production of poly -β- hydroxybutyrate by Azotobacter chroococcum was enhanced up to 80% of the cell dry weight after 48h, at 120 rpm and 30°C. P(3HB)was purified and identified by comparing the data with published one. Selection P(3HB) degrading actinomycetes was done by clear zone method, the wider clear zone organism was selected. Streptomyces violaceus was the best organism for degradation P(3HB) after 12 day. Optimization of culture condition for P(3HP) degradation are discussed. Lipase produced by Streptomyces albidoflavus cannot degraded P (3HB).

21/11 EVALUATION OF GEN-PROBE AND PCR FOR DETECTION OF CHLAMYDIA TRACHOMATIS IN URINARY TRACT INFECTIONS, PROSPECTIVE STUDY.

*M. El-Sayed,**W. Badawy, A. Bakr

*Clinical pathology, **Microbiology, Urology Centre, Faculty of Medicine,Mansoura University, Egypt

A total of 50 patients (20 males and 30 females) were enrolled in this study. The patients' complaints were upper or lower urinary tract infections with sterile pyuria. Routine bacterial culture were done to evaluate absence of gonococci and other bacteria. Samples were 15 ml of urine. Well mixed urine samples were divided   into 4 aliquots. One aliquot was used for chlamydia culture. Other sample was used for Gen-probe. Two aliquots were tested for chlamydia antigen by ELISA and Chlamydia DNA by PCR. In the studied patients, chlamydia culture was positive in 25 patients (50%).In comparison of chlamydia antigen detection by ELISA versus culture, the sensitivity of ELISA was 40% , specificity was 100%, accuracy was 70%, positive predictive value was 100% and negative predictive value was 37.5%. In study of Gen-probe versus culture, the sensitivity of Gen-probe was 100%, specificity was 40%, accuracy was 70%, positive predictive value was 62.5% and negative predictive value was 100%. In study of PCR versus culture, the sensitivity of PCR was 60%, specificity was 100%, accuracy was 80%, positive predictive value was 100% and negative predictive value was 71.4%. From this study it could be concluded that C. trachomatis should be considered as a possible etiological agent in upper urinary tract infections as in lower urinary tract infections. Diagnostic strategies should be targeted for its possible presence. Molecular biological techniques either non amplification (Gen-Probe) or with amplification (PCR) are valuable tools in laboratory diagnosis of C .trachomatis. ELISA technique with its marked reduced sensitivity could not be used as a   screening tool.

22/11 FERMENTATIVE PRODUCTION OF GIBBERELLINS BY ASPERGILLUS NIGER

A.S. Gad

Chemistry of Natural and Microbial Products Lab., NRC, Dokki, Giza, Egypt.

The production of gibberellic acid by A.niger was found to be influenced by some factors. The studied nutritional factors were concentration of both glucose and nitrogen, effect of exogenous addition of sodium acetate or MgSo4.5H2o.,the culture pH of submerged or surface liquid culture. The yield of gibberellic acid was improved by judicious selection of these parameters, thus the increase in gibberellic acid yield in the molasses medium (340 mg/L) was acheived by using 3g/L sodium acetate as a direct precursor to gibberellic acid production. pH 4-5 seemed to be favourable on using 160 g/L molasses for 9 days at 30ْ C.

 


23/11 INFLUENCE OF POLYACRYLIC ACID POLYMERS (CARBOPOL 940) ON THE ANTIPSEUDOMONAL ACTIVITY OF GENTAMICIN AND CIPROFLOXACIN

M.A. Fawzy and E. Aboulmagd

Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.

The bacteriostatic and bactericidal activity of gentamicin and ciprofloxacin was determined in the absence and presence of 0.1% carbopol 940 (polyacrylic acid polymer), using the broth macro dilution technique against six Pseudomonas aeruginosa clinical isolates. The MIC (minimum inhibitory concentration) and the MBC (minimum bactericidal concentration) values of gentamicin and ciprofloxacin were remarkably increased in the presence of 0.1% Carbopol 940 which reflect the inactivating action of carbopol on the antibacterial activity of the tested antimicrobial agents. In addition, 0.1 % carbopol antagonized the bactericidal activity of gentamicin and ciprofloxacin (½, 1 and 4xMIC) studied by the time–killing method. The bactericidal activity of gentamicin (4xMIC) was completely removed in presence of 0.1% carbopol while that of ciprofloxacin was partially inhibited. In conclusion, The effect of carbopol 940 must be taken in consideration not only when formulating fluoroquinolones and aminoglycosides in presence of carbopol but also in the coadministartion of these antibiotics with other medications containing carbopol (e.g. eye drops).

 

24/11 USE OF PROPOLIS AS A PRESERVATIVE TO FETA CHEESE MANUFACTURED BY ULTRAFILTRATION TECHNIQUE

M.M. Ashour, A.A. Abd El-Baky, A.H. Guirguis and E.M. Abd El-Wahed

Food Science Department, Faculty of Agriculture, Zagazig University

In studying the effect of propolis on the properties of Feta cheese, eight levels from the 10% aqueous solution of propolis i.e. (2.5, 5, 7.5, 10, 12.5, 15, 17.5, and 20 ml) per liter of the milk retentate. This was compared with Feta cheese without propolis. The extract was added to the milk retentate, then the retentate transferred to Feta cheese and stored at 5±2ºC. The sensory evaluation, microbiological and chemical analysis was carried out on the following periods: fresh, 7 days, 14 days, 21 days and 28 days. The moisture, salt, acidity were higher than the control. Fat percentage was lower than the control. Nitrogen fractions in some treatments were higher than that of the control, but TVFA was lower than the control. It was clear that the propolis was highly effective on the yeast and mold count specially at the higher concentrations during storage. It was observed that the molds were vanished completely in the treated cheese. The propolis was helped in prolonging the shelf life of Feta cheese affected as a preservative. The propolis was able to destroy most of the undesirable microbes which have a bad effect on the human health.


25/11 SERUM LEVELS OF TUMOR NECROSIS FACTOR-ALPHA AND TRANSFORMING GROWTH FACTOR-BETA IN HEPATITIS C VIRUS INFECTION: CORRELATION WITH VIRAL LOAD AND LIVER ENZYMES

A A..Ghazal and A. E. Abd El-Wahab*

Departments of Microbiology, Medical Research Institute, Alexandria University, *Medical Biotechnology, Genetic Engineering & Biotechnology Research Institute (GEBRI), Mubarak City for Scientific Research & Technology Applications, Alexandria.

Tumor necrosis Factor alpha TNF-α, is an immunoregulatory cytokine with a wide range of biological activities. Transforming growth factor betaTGF-β1 is a pluripotent cytokine that promote hepatic fibrogenesis. The present study measures serum levels of both TNF-α and TGF-β1 in patients with HCV and correlates the level of these cytokines with both viral load and liver enzymes. Forty-five anti-HCV positive patients together with ten controls were enrolled in this study. Anti-HCV positive patients were tested for the presence of HCV-RNA by RT-PCR. Quantitation of HCV-RNA was performed for HCV-RNA positive subjects using real time quantitative RT-PCR. Serum levels of TNF-α and TGFβ1 were measured by enzyme linked immunosorbent assay and biochemical determinations of liver enzymes were done for all cases. The 45 anti-HCV patients were further divided into 15 HCV-RNA negative patients and 30 HCV-RNA positive patients. Liver enzymes were more elevated in HCV-RNA positive group than in HCV-RNA negative group and this difference was statistically significant (For ALT, P<0.01, GGT P<0.001, AST P<0.01). Aminotransferases did not correlate with viraemia, while GGT correlated with viraemia (P<0.001). Serum TNF-α levels were significantly elevated in HCV patients (P<0.01) and significantly correlated with the presence of viraemia P<0.001. Serum TGF-β1 levels were not different in relation to viraemia. Both TNF-α and TGF-β1 did not correlate with aminotransferases, while TNF-α correlated with GGT. This data indicate that TNF-α and GGT could reflect liver injury and may be affected by viral activity.

 

26/11 NUTRITIONAL UPGRADING OF SOME AGRICULTURAL RESIDUES BY PHANEROCHAETE CHRYSOSPORIUM

A.S. Hamza, G.A.M.A. Darwish and T.F. Mohammedi

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

White rot fungi (Phanerochaete chrysosporium) was grown on tomato pomace, lentil straw and olive branches or leaves individually or in combination mixture (1:1) using solid state fermintation technique. Agricultural residues containing lignocellulose can be up-graded by white rot fungi using solid-state fermentation. Lignocellulosic materials were treated with 12% of cellulolytic and ligninlolytic fungal inoculum and incubated at 35oC for 30 days. In natural decomposition of lignocellulosic matter, white rot fungi paly an important role for degrading holocellulose and lignin to lower-molecular weight products. Phanerochaete chrysosporium succeeded to increase crude protein of lentil straw from 6.90% to 10.80% and tomato pomace from 10.01% to 11.65%. In addition, the tested fungus increased crude protein of combination mixture (1:1) of tomato pomace and olive branches or leaves from 9.70% to 14.43%. Protein of combination mixture of olive branches or leaves and lentil straw (1:1) increased from 7.20% to 10.82%, while, tomato pomace and lentil straw (ratio 1:1) increased from 9.10% to 11.40%. A decrease ranged from 25 to 30 of legnin content was determined.

 

 

27/11 UTILIZATION OF SPAWN PREPARED ON DIFFERENT FOOD INDUSTRIAL WASTES FOR PRODUCTION OF SOME SPECIES OF PLEUROTUS SPP. WASTES.

G.A.M.A. Darwish and K.A. Abdelkawi*

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

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

Giza, Egypt

Huge amounts of agricultural crop residues are generated in Egypt every year, mainly rice straw, maize stalks and cotton stalks, as well as another huge amount of food industrial wastes, most of these residues are composed of three major groups of polymers, cellulose, hemicellulose and lignin. Four food industrial wastes namely, orange peels, mandarine peels, pea straw and potato jacket were used as carrier for spawn preparation. The best spawn were prepared on combination mixture between mandarine peels and rice straw at ratio 1:1 and orange peels and rice straw at the same ratio. Two strains of Pleurotus were cultivated on rice straw, maize stalks and cotton stalks at spawn ratio 5% also, the above mentioned three agricultural wastes were supplemented with rice bran in three different ratios 0, 3, 5 and 7% w/w. The obtained results indicated that there are no significant differences between 5 and 7% rice bran additives. The biological efficiency (B. E%) at 5% rice bran additives were 62.5, 49 and 30% for P. flabellatus grown on rice straw, maize stalks and cotton. While B. E% for control were 46.5, 37 and 29%, respectively.

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