Vol. 4, January, 2003

faux kamagra 1/4 IMMUNOBLOT AND REVERSED PASSIVE LATEX AGGLUTINATION FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXINS

A. Atia

Department of Microbiology and Immunology,

Faculty of Vet. Medicine, Zagazig University

The production of staphylococcal enterotoxins   (SE) , types (SEA, SEB, SEC,, SED) and toxic shock syndrome toxin (TSST) was studied in 31 and 26 identified strains from raw cow’s milk and chicken’s skin respectively. The characterized enterotoxigenic strains that isolated from skin and milk represented 43% and 13% of total isolates, respectively. The strains producing enterotoxins were biotyped as S. aureus, S. cohnii, S. epidermidis, S. hominis and S. gallinarum. The enterotoxigenic isolated staphylococcal strains showed higher tendency to produce DNase and coagulase. Out of 15 enterotoxigenic tested isolates, 11 (73.3%) were DNase – positive and 13 (86.6%) were coagulase – positive. Only SEC, SEB and TSST were produced and secreted to the culture medium, in addition to simultaneous production of the enterotoxins C and B by some isolates . The enterotoxigenicity was detected by reversed passive latex agglutination (RPLA), polyacrylamide gel electrophoresis stained with silver stain and western immunoblot techniques. The strains positive by RPLA were also positive silver stained polyacrylamide gel and western immunoblot.

achat priligy en france 2/4 CHARACTERIZATION OF PSEUDOMONAS AERUGINOSA AND ITS MULTIDRUG-RESISTANT MUTANTS IMPLICATED IN CHRONIC SUPPURATIVE OTITIS MEDIA

H.A. Abdel-Salam, A. Atia* and M.M. Aboulwafa**

FROM

Department of Microbiology and Immunology, Faculty of Pharmacy and

Veterinary medicine, University of Zagazig* and **Ain-Shams, Egypt

Development of prix du viagra au québec Pseudomonas aeruginosa mutants in chronic suppurative otitis media (CSOM) hinders the treatment and essentially requires coverage with more than one antibiotic. In addition to the original infective sortie generique viagra P. aeruginosa serogroup O:11 isolate, seven drug-resistant mutants against amikacin (AK), amoxycillin/clavulanic acid (AMC), cefoperazone (CFP), ofloxacin (OFX), piperacillin (PRL), rifampicin (RD), and tobramycin (TOB) from the exudates of CSOM were isolated and characterized. Plasmids analyses of these mutants showed significant difference in sizes and functions. The original isolate and AMC-mutant showed plasmid of about 7.5 kb, while AK, CFP and RD-mutants were harbored the largest plasmids of about 18 kb. OFX-mutant showed the smallest plasmid of about 5.5 kb and PRL-mutant has a plasmid of 8.5 kb. TOB-mutant contained 2 plasmids of sizes about 8.5 and 12 kb. Plasmids were cured by heat, ciprofloxacin and acridine orange. Heat was the most efficient plasmid curing agent followed by ciprofloxacin then acridine orange. All the mutants were resistant to RD even after plasmid curing, except TOB-mutant cells were sensitive to RD before and after plasmid curing. In contrast, the mutants were sensitive to PRL, except TOB-mutant cells were resistant to PRL either plasmid cured or uncured. Moreover, the mutants showed different and irregular patterns of susceptibility to the remaining tested antibiotics. Drug combinations revealed that both piperacillin and ofloxacin were synergistic against the detected original achat viagra en thailande P. aeruginosa isolate and its mutants. It is concluded that CSOM with achat cialis moins cher P. aeruginosa harbored different mutants that might be acquired different plasmids or DNA fragment under the infection conditions. Culture and antibiotics sensitivity tests were recommended and the implications of frequent mutants required suitable combined antibiotics therapy.

 

3/4   KINETIC OF STREPTOMYCIN PRODUCTION AND DEGRADATION DURING BATCH CULTIVATION OF STREPTOMYCES GRISEUSIN SHAKE FLASK AND BIOREACTOR

H.A. El-Enshasy; K.Y. El-Shahed* and H.M. Mohammed*  

Bioprocess Development Dept. Genetic Engineering and Biotechnology Research Institute, Mubarak City for Scientific Research, New Burg-Al-Arab, Alexandria, Egypt.

* Natural and Microbial Products Dept., National Research Center, Tahrir Street, Dokki, Cairo, Egypt.

In the present investigation, the kinetics of streptomycin production and degradation by quand le cialis generique Str. griseus were studied in details in both shake flask and bioreactor cultures. Both of volumetric and specific antibiotic production were higher in case of bioreactor culture by about 38% and 66%, respectively. On the other hand, the cell growth was higher in shake flask culture and reached 8.2 g/l (about 93% higher than the biomass in the bioreactor culture). After glucose limitation, degradation of antibiotic accompanied with a significant decrease in cell dry weight was observed in both cultures. The antibiotic degradation rate and the cell lysis rate were higher in bioreactor culture.

cialis périmé risques 4/4 PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF AN ALKALINE SERINE PROTEASE OF TWO HIGHLY MOSQUITOCIDAL STRAINS OF BACILLUS SPHAERICUS AND POSSIBLE APPLICATIONS.

M.E. Moharam, M.A. El-Bendary and M.S. Foda

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

Alkaline serine protease [EC 3.4.21] of two highly mosquitocidal strains of quelle dose de levitra prendre Bacillus sphaericus ( ou trouver du cialis en pharmacie B.s.) namely the international strain 2362 (IS 2362) and a local Egyptian strain NRC 69 was purified to homogeneity and characterized. The two enzymes were purified 58 and 126 folds with specific activity of 75 and 168 units/mg protein for strains IS 2362 and NRC 69 respectively. The purified enzymes of the two sources exhibited high degree of similarity showing maximal activity at pH 8.5 at 60°C reaction temperature. Michaelis constants (Km) values for azocasein substrate were 0.3 mg/ml for B.s. IS 2362 and 0.2 mg/ml for B.s. NRC 69. The enzymes were thermostable up to 50°C. Calcium chloride (2mM) stabilized the enzymes upon heating at 55°C for 60 minutes thus retaining between 73% and 80% of their activity as compared to less than 50% in the absence of the salt. Calcium, magnesium and cobalt ions exhibited little or no effect on enzyme activity. Ferrous, zinc and mercurous ions exerted clear inhibitory effects at 2 mM final concentration. The alkaline protease was stable in the presence of 500 mg chlorine/l and in the presence of laundry detergents. It was efficiently hydrolyzed the gelatin layer of used X-ray films within 4 and 7 min at 65 units protease /ml and 32.5 units protease/ml respectively for alkaline protease produced by both B.s. strains.


5/4 UTILIZATION OF SOYBEAN WASTE FOR PRODUCTION OF AMYLASE AND CELLULASE ENZYMES BY EDIBLE MUSHROOMS

H.M. Abdou

Biochemistry Department, National Research Center, Tahrir st., Dokki, Cairo, Egypt.

Soybean waste, produced in large amounts during soybean processing, was utilized by Lentinus edodes NRRL 22663 and Pleurotus ostreatus NRRL 0366 for amylase and cellulase production. The most favourable culture conditions affording the highest Pleurotus ostreatus amylase and cellulase activities were achieved by growing it on media containing 10% dry soybean waste and 0.2% asparagine. The best one for L. edodes amylase production were 10% dry soybean waste and 0.2% ammonium sulphate, while for L. edodes cellulase 0.2% asparagine and 5% dry soybean waste were the best. The harvest mycelia of P. ostreatus was 3.4 g dry weight/L, and for L. edodes was 3.3 g dry weight/L after 9 days incubation at 28°C and on static condition and air/medium ratio (4 : 1). Partial purification of the two enzymes was achieved by fractional precipitation with ammonium sulphate. The best specific activities of P. ostreatus and L. edodes amylase were obtained with 60% (NH4)2SO4 concentration. While 80% ammonium sulphate concentration was the best with P. ostreatus and L. edodes cellulase. Saccharification of some pretreated lignocellulosic wastes using crude P. ostreatus and L. edodes cellulase and amylase showed promising results.

6/4 TRANSMISSION OF HEPATITIS C VIRUS BETWEEN SPOUSES

Z. El Kholy*, T. Moghazi**, N. Abd El Hakeem* and A. Ghazal ***

Departments of Applied Medical Chemistry*, Clinical Pathology **, Microbiology***. Medical Research Institute, Alexandria University, Egypt.

This study was carried out to elucidate the role of sexual transmission of HCV. One hundred anti-HCV positive individual (Index cases) and their sexual partners were submitted to detailed questionnaire. Sexual partners of anti-HCV positive patients were examined for anti-HCV. HCV-RNA by RT nested PCR and liver enzymes (ALT and AST) were determined for both anti-HCV positive index cases and their anti-HCV positive partners. The present study revealed that the prevalence of anti-HCV among sexual partners (14%) was similar to that of the blood donors (14.78%) attending the blood bank of the Medical Research Institute, Alexandria, Egypt. HCV infection risk increased by 2 fold with the duration of marriage. HCV transmission was significantly more efficient from women to men 30.76% compared to 8.1% from males to females. Among our group studied, HCV viremia was not an increased risk for developing HCV infection. Ten out of the fourteen anti-HCV positive sexual partners had other potential HCV risk factors other than sexual transmission while four had no obvious risk for HCV infection, as their sexual partners were viremic and they may acquire the HCV infection through their partners. Sequence of nucleotides of HCV genome of both partners would be a strong evidence of sexual transmission.


7/4 THE BACTERIAL FLORA IN THE RHIZOSPHERE OF SOME HALOPHYTES EXIST IN EGYPT

M. Abedel Fattah

Department of Botany, Faculty of Science, Beni-Suef 62511, Egypt

The eubacterial flora in the rhizosphere of five halophytes were isolated and characterized in a salt-containing medium. The halophytes were collected from desert lands, cultivated reclaimed lands, cultivated salt-affected lands, reclaimed salt-affected lands and Qaroun lake shore in Fayoum area. The majority of the bacterial flora were tolerant to salinity up to 15% NaCl, but about 39 % and 26 % of the isolates persisted at 20 % and 25 % NaCl, respectively. Extremophiles (halophilic bacteria that are not able to grow at non-saline medium) were also isolated, they represented about 8 % of the bacterial isolates. The bacterial flora were mainly mesophilic, however, 28 % and 21% of the isolates were thermotolerant, being grew at 60oC and 65oC, respectively. The bacteria exhibited a wide pH range, they grew in the slightly acidic medium (pH 4 and pH 5) as well as in the highly alkaline medium (pH 9, pH 11 and pH 12). The bacteria were tentatively identified and classified based on phenotypic characteristics (e.g., cell morphology, antibiotic sensitivity and biochemical activities), and after numerical analysis. Spore-forming gram positive rods were assigned to five species of the genus Bacillus. The cocci were related to the genera Staphylococcus and Planococcus. The gram   negative rods represented about 26 % of the isolates, they ranged from a very small rods and coccoids to the elongated rods. The strict halophilic (small gram negative rods) bacteria were related to the genus Vibrio. Other halophilic gram negative bacteria were related to the genera Azotobacter, Rhizobium and Pseudomonas. It is concluded that the halophytes live in association with certain haloalkaliphilic bacteria of distinguished ecological characters.

 

8/4 ALLELOPATHIC POTENTIAL OF SUNFLOWER TOWARDS MAIZE RHIZOSPHERE MICROFLORA INFESTED WITH CEPHALOSPORIUM MAYDIS SAMRA

M.A. Rizk and T.A.A. Moussa

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

In this study, the authors investigate the allelopathic potential of sunflower residues on the emergence and infection percentages of maize plant when planted in infested soil with the late wilt fungal pathogen Cephalosporium maydis, also the effect of sunflower residues on rhizosphere and non-rhizosphere microflora. The emergence percentage of maize seedlings were slightly inhibited with sunflower treatments, but showed improvement in the emergence in presence of the pathogen. The addition of sunflower residues to the soil adversely affected the microflora of maize; but Fusarium verticillioides not affected by any treatment of sunflower residues. The study also revealed that the most common fungal species were Aspergillus flavus, A. terreus, Fusarium solani and F. verticillioides in addition to maize pathogen C. maydis.


9/4 INHIBITORY EFFECT OF CHITOSAN AND ITS DERIVATIVES ON SOME PHANT VIRUSES

S.Y.M. Mahmoud; K.A. El-Dougdoug*; M.H. Abdel-Ghaffar* and A.S. Aly**

Department of Agricultural Botany, Faculty of Agriculture, South Vally University, Sohag, Egypt.

*Department of Agricultural Microbiology (Virology Lab.), Faculty of Agriculture, Ain Shams University, P.O.Box 68 Hadayek Shubra 11241, Cairo, Egypt.

**Textile Research Division, National Research Centre, Dokki Cairo, Egypt

In this study, chitosan (poly 1-4- b, D- glucosamine) and its derivatives (carboxymethyl, glutamate and citrate chitosan) were used as anti-plant viruses substances. The inhibitory effect of chitosan was pronounced either using it by spraying or adding to the viral inoculum. However, the addition of chitosan to the viral inoculum was more effective than spraying it. The results indicated that the chitosan was generally more effective than its derivatives on tomato mosaic virus (ToMV) even at lower concentrations (0.0001%). Chitosan had ability to inhibit the virus infection even at high concentration of purifed virus. Electron micrographs of ToMV showed presence of non-rigid, thick and quasi-malformed of rod virus particles aggregates, when the purified preparations treated with 0.12% chitosan solution. Datura stramonium plants were exhibited systemic acquried resistance (SAR) to ToMV infection, when their lower leaves treated with 0.12% chitosan solution pre-mechanical inoculation with ToMV. Moreover, it was found that the systemic infection caused by ToMV, bean yellow mosaic virus (BYMV), cucumber mosaic virus (CMV) and potato virus Y (PVY) were reduced with various degrees when their systemic hosts treated with 0.12% chitosan solution pre-viruses mecanically inoculated. The banana bunchy top virus (BBTV) and banana-CMV were not detected using DAS-ELISA in any resulting parts, i.e., leaves, midribs, roots and corms, when their mother corms treated with 0.12% chitosan solution

 

10/4 PUTATIVE EXTRACELLULAR VIRULENT LIPOLYTIC ENZYMES SECRETED BY PATHOGENIC CANDIDA ALBICANS

M.M.Kh. Okasha

Department of Microbiology, Faculty of Pharmacy, Zagazig University, Egypt.

The role of extracellular lipase, phospholipase and esterase in the pathogenicity of Candida albicans was investigated by comparing 20 pathogenic isolates from different infected sites (cadidaemia, vaginitis, buccal moniliasis and tineasis) with equal number of commensal strains isolated from the same sites of healthy volunteers. All isolates were screened for lipolytic activity on egg-yolk agar as traditional screen with triglycerides substrate or Czapek-Dox tributyrin medium (CDTM) for lipase or with phospholipids substrate for phospholipase activity and analyzed spectrophotometrically for lipase and phospholipase concentration and on 0.7% yeast nitrogen base with 2.5% tween 80 for esterase and assayed colorimetrically. Pathogenic isolates produced significantly more extracellular phospholipase (from 2 to 7 folds by hydrolytic activity and 27 to 37 folds spectrophotometry); lipase and esterase activity than the commensal strains did. The exhibited activities of the produced enzymes among the groups were in the following rank: fungaemia, vaginitis, moniliasis, tineasis (100%) and finally few commensals (22.4%). The isolates were also characterized for their lipase, phospholipase and esrerase production and growth. These showed the secretion of lysophospholipase-transacylase in addition to phospholipase. Lipase was detected only when C. albicans was grown in media containing tween 80, other tweens or triglycerides as the sole carbon source and not in Sabouroud Dextrose broth or yeast/ peptone/dextrose media. Southern blotting analyses revealed that C. albicans may contain a lipase gene family. On the other hand, Western blot analysis of phospholipase in culture supernatants using anti-phospholipase polyclonal sera, detected phospholipase only in concentrated filtrates. Additionally, carbohydrates supplementation inhibited or at least did not induce lipase expression. Extracellular esterase activity was detected only in presence of tween 80 as a sole carbon source and maximized at p H 5.5 to below 6. With the exception of tween 80, tween 20, sod.taurocholate (NaTC) and taurodeoxycholic acid (TDCA) which potentiated esterase activity, various other tested reagents had no significant effect. Thin – layer chromatography revealed that this enzyme does not hydrolyze triolein and l-α lecithin suggesting that it is a monoester hydrolase. It was suggested that lipase, phospholipase and esterase may well contribute to the pathogenicity of C .albicans by abetting the fungus in damaging and traversing host cell membranes, processes which likely increase the rapidity of disseminated infection.

 

11/4 EVALUATION OF THE EFFECT OF DIFFERENT ANTIMICROBIAL AGENTS ON ENDOTOXIN RELEASE FROM ESCHERICHIA COLI AND PSEUDOMONAS AERUGINOSA

S.M. Kheira and M.I. Awad

Microbiology Dept., Faculty of Pharmacy, Mansoura University,

Mansoura, Egypt.

The effects of different antimicrobial agents including ceftazidime, imipenem, cephaloridine, gentamicin and ofloxacin on endotoxin release were evaluated in vitro on clinical isolates of Escherichia coli and Pseudomonas aeruginosa. The amount of endotoxin released was measured by chromogenic limulus amoebocyte lysate assay. The corresponding viable cell counts were also determined by pour plate method. Higher degree of reduction in endotoxin release was observed with imipenem followed by ceftazidime from the tested isolates. The degree of reduction in case of cephaloridine was concentration-dependent, while it was strain-dependent with gentamicin and ofloxacin. Although the obtained results revealed that imipenem and ceftazidime are suggested as antimicrobials of choice in treatment of septic infections due to Gram negative rods, further clinical investigations are necessary to verify these crucial findings.

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