Vol. 30, September, 2011.

cialis douleurs dorsales 1/30 OPTIMIZATION OF CULTURE PARAMETERS FOR PRODUCTION OF ALKALINE PROTEASE FROM ALKALIPHILIC-HALOTOLERANT STRAIN OF PENICILLIUM CHRYSOGENUM

Mohamed Ghareib and Medhat Abu Tahoon

Department of Biological and Geological Sciences, Faculty of Education,

Ain Shams University, Roxy, Cairo, Egypt

Alkaliphilic- halotolerant strain of viagra et troubles cardiaques Penicillium Chrysogenum was found suitable for production of detergent enzyme complex containing an alkaline protease. The fungus grew optimally at pH 9 in presence of 7.5% NaCl (w/v). Its enzyme complex contained alkaline activities of protease, a-amylase and cellulase with prevailing of the first enzyme. The culture parameters for optimum production of alkaline protease were followed in this work. Optimum protease expression occurred in presence of 0.9% sucrose and 0.7% casein after 7 days of static incubation at 30oC when the fermentation medium was adjusted to pH 9. The crude alkaline protease of prix du kamagra en gel P. chrysogenum was characterized in terms of pH, thermal activity and stability. Maximal activity was recorded at pH 9 and 40oC. The enzyme was found to be alkaline thermotolerant by retaining 85% and 59% of its original activity after exposure for one hour to pH 9 and 70oC, respectively.

cialis 20 mg pour femme 2/30 ASSESSMENT OF THE INHIBITORY EFFECT OF PROPOLIS ON HUMAN COLON CARCINOMA (HCT116) AND HEPATOCELLULAR CARCINOMA (HEPG2) CELL LINES AND ITS IMPACT ON APOPTOSIS

Abeer A. Bahnassy, Maha H. Elazizy*, Amany A. Tohamy**, Abdel-Rahman N. Zekri* and Waleed S. Mohamed*

Pathology,*Cancer Biology Departments, National Cancer Institute, Cairo University and **Zoology Department, Faculty of Science, Helwan University.

Several studies have reported that Propolis obtained from honey bee hives has anti-mutagenic and anti-carcinogenic properties with biological and therapeutic effects. The present study was performed to investigate the potential cytotoxic effect of propolis on human colon carcinoma cell line (HCT116) and hepato cellular carcinoma (HEPG2) cell line and its possible apoptotic effect. Egypt propolis was used at a concentration range from5 µg/ml to30 µg/ml, at different time intervals (24, 48, 72 and 96 h). The ratio expression of FAS and FAS-L genes was assessed using semi-quantitative reverse transcriptase-PCR technique and flowcytometric analysis to evaluate the apoptosis of cells treated with propolis. Our study showed that propolis had a strong cytotoxic effect on both HCT116 and HEPG2 cell lines, and triggered apoptosis at a dose and time-dependent manner. This finding may be important economically and allow development of a relatively inexpensive cancer treatment.

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acheter du viagra en pharmacie sans ordonnance forum 3/30 EVALUATION OF PHENOTYPIC AND GENOTYPIC METHODS FOR DETECTION OF METALLO-Β-LACTAMASES (MßLS) PRODUCTION AMONG CARBAPENEM-RESISTANT STRAINS IN MDR PSEUDOMONAS AERUGINOSA URINE ISOLATES.

Tharwat R. El-Khamissy, Ehsan Abdel Saboor*, Abdel Fatah I. Ahmed**,

Salwa Seif El-dein* and Bahaa Anwer

Microbiology & immunology Dep. (Faculty of Pharmacy, Al-Azhar University-­ Assiut); *Faulty of Medicine, Assiut University; **Urology Dep., Faculty of Medicine, Assiut University.

Carbapenem-resistant Pseudomonas aeruginosa resulting from metallo-ß-lactamases (MßLs) has been reported to be an important cause of nosocomial infection and is a critical therapeutic problem worldwide. The aim of the work was to determine the prevalence of carbapenem-resistant among multidrug-resistant (MDR) P. aeruginosa urine isolates, as well as to compare the different phenotypic and genotypic methods for detection of MßLs positive strains among carbapenem-resistant MDR P. aeruginosa isolates and to detect antimicrobial agents effective against to such isolates. The study was conducted over a period of 12 months from September 2008 to October 2009 on 1337 catheterized patients that admitted Urology Department at Assiut Teaching Hospital. 23 carbapenem -resistant P. aeruginosa strains were isolated from urine samples of catheterized patients admitted Urology Department and the isolated strains were tested for MßLs production by phenotypic and genotypic methods. One hundred and fifteen P. aeruginosa isolates (17.7%) were isolated from urine samples of catheterized patients. Twenty-three (20%) of P. aeruginosa isolates were carbapenem-resistant. Eight and thirty (33.04%) P. aeruginosa isolates were multidrug resistant (MDR), and 66.96 % of P. aeruginosa isolates were non-multidrug resistant (non-MDR). The prevalence of imipenem- and meropenem-resistant strains among MDR P. aeruginosa strains was 90% and 82.61% respectively. Positive phenotypic detection for MßLs production among carbapenem-resistant MDR strains ranged from 57.89-89.47%, and Polymerase Chain Reaction (PCR) for detection of bla IMP-1 and bla VIM-1 gene were positive in 68.43% of strains (of note, 26.32% of these strains were positive to bla IMP-1 gene and 42.11% were positive to bla VIM-1 gene). Carbapenem -resistant MDR P. aeruginosa strains showed high degree of resistance to most antipseudomonal agents. However, bothpolymyxin-B andcolistin demonstrated good activity 88.89% and 83.33% respectively, followed by tobramycin (44.44%), both levofloxacin and piperacillin/tazobactam combination, for each one (39.89%), and cefepime (33.33%). It conclude that the best method for screening of MßL production in P. aeruginosa strains was imipenem-EDTA combined disc test and imipenem-EDTA MßL E test both were equally effective for MßL detection, however, we believe that the cost-constrains, combined disc test could be used as a convenient screening method in the clinical laboratories.

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trace de viagra dans le sang 4/30 THE RELATION BETWEEN CHLAMYDIA PNEUMONIAE INFECTION AND OTHER RISK FACTORS IN PATIENTS WITH CORONARY HEART DISEASE

Abeer A Ghazal, Dalia El Sayed Metwally and Kamal Mahmoud*

Departments of Microbiology and *Cardiology and Angiology, Medical Research Institute, Alexandria University.

Coronary heart disease (CHD) is a serious public health problem. It is the third major cause of death in developed countries. sur quel site acheter du viagra Chlamydia pneumoniae ( qual e o melhor cialis ou levitra C. pneumoniae) is a cause of respiratory tract infection worldwide and recently it has been implicated as a possible cause of atherosclerosis and coronary artery disease. The current study was undertaken to investigate the possible association between infection with C. pneumoniae and coronary heart diseases (CHD) (as determined by the presence of C. pneumoniae specific serum IgG antibodies, chlamydial DNA in PBMNC by polymerase chain reaction), and to study the relation between C. pneumoniae and other risk factors for CHD. Our results showed a significant increase in IgG seropositivity against C. pneumoniae among CHD patients compared to the control group   (86%, 40%, respectively, p<0.05). The carriage of C. pneumonia DNA in PMNCs was significantly higher in coronary artery disease (CAD) patients than controls (44%, 0% respectively, p<0.05). Age and sex were not associated with the presence of C. pneumoniae DNA in peripheral blood mononuclear cells (PBMNCs) of CAD patients; also there was no significant association between diabetes mellitus, hypertension, and smoking and the presence of C. pneumoniae DNA in PBMNCs of CAD patients. The presence of diabetes mellitus, hypertension, and smoking were significant risk factors in CAD patients. Total cholesterol, triglycerides, LDL-C, fibrinogen and hs-CRP levels were significantly increased in CAD patients as compared to controls.

 

5/30 EMERGENCE OF EXTENDED SPECTRUM BETA LACTAMASES IN CLINICAL ISOLATES OF PROTEUS MIRABILIS IN ISMAILIA, EGYPT

Samira Zakeer, Nora Fahmy Mahmoud, Alaa El Din. M. S. Hosny

and Salah Abd Allah

Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University and Cairo University.

Extended spectrum Beta lactamases (ESBLs) producing gram negative bacteria is an increasing problem worldwide. A total of 30 Proteus mirabilis isolates were collected from the Ismailia Specialist Laboratory and the Educational Hospital of Suez Canal University. Those samples were analyzed for the production of ESBLs phenotypically. ESBL-production was detected in 26.6% of the isolates.


6/30 ANTIBIOGRAM, PLASMID PROFILE AND MOLECULAR CHARACTERIZATION OF UROPATHOGENIC ESCHERICHIA COLI ISOLATES

Amal E. Ali

Department of Microbiology and Immunology, Faculty of Pharmacy,

Cairo University

Uropathogenic Escherichia coli isolates (35) obtained from a private medical analysis laboratory, Cairo, Egypt, were characterized with regards to antibiotic resistance, plasmid profile and molecular pattern. Isolates were sensitive to imepenum and were resistant to ampicillin and most of them were multidrug- resistant. One to 4 plasmids were detected in most of the isolates. One common plasmid being identified in all plasmid –harbored E. coli strains. Seven plasmid profiles were obtained from 25 plasmid containing strains. The prevalence of the virulence factors was 42.8%, 8.5%, 8.5%, 60.0%, and 14.2% for pap (fimbriae adhesin), sfa (fimbriae adhesin), vat (vacuolating autotransporter toxin) hem (hemin importer) and hlt (heat labile toxin) respectively when assayed using specific primers in PCR reactions.

 

7/30 INFLUENCE OF SOME BIOLOGICAL OR PROLINE TREATMENTS ON SUGAR BEET UNDER NITROGEN LIMITATION IN SALINE-SODIC SOIL

Faten M. Mohamed, Soha S.M. Mostafa and Nabil A. Omar

Microbiol. Res. Dept., Soils, Water and Environ. Res. Inst., Agric. Res. Center (ARC), Giza, Egypt.

The effect of exogenous amino acid L- proline was compared with the effect of biological inoculation (cyanobacteria and/or N2-fixing bacteria) on sugar beet yield and yield quality in saline-sodic soil. A field experiment was conducted at Sahl El-Hussinia Res. station, El-Sharkia Governorate, Egypt under two levels of mineral nitrogen fertilization (50 and 75% of nitrogen recommended dose). Soil enzyme activities (dehydrogenase and nitrogenase), total bacterial counts and total cyanobacterial counts were significantly maximized by the combined inoculation of cyanobacteria and N2-fixing bacteria along with 75% N. While, the individual N2-fixing bacterial inoculation resulted in the significantly highest records of soil N2-fixing bacterial counts combined with 75% N compared to proline treatment and controls. The availability of P, K and N (in the form of NO3- -) was significantly maximized when applying the combined inoculation of cyanobacteria and N2-fixing bacteria combined with 75% N comparing with proline treatment control plots. On the other hand, the availability of Na in soil was significantly decreased by all the biological and proline treatments comparing with control plots. All treatments had no significant effect on soil pH values. While, the combined inoculation (cyanobacteria and N2-fixing bacteria) led to the most significant decrease of soil EC (1:5 soil water extract) values along with 75%N comparing with proline treatment and control plots. Proline and the biological inoculation showed a significant positive impact on some physiological properties of plants drown at 75% nitrogen level, such as chlorophyll in leaves, proline and phenolic compounds in roots. The highest responses for these traits were in proline-treated plots followed by the combined inoculation of cyanobacteria and N2-fixing bacteria. However, proline-treated and the combined-inoculated (cyanobacteria and N2-fixing bacteria) plots recorded the highest similar significant root yield productivity along with 75% nitrogen fertilizer. The combined inoculation increased positively N, P and K uptake and decreased the uptake of Na in sugarbeet roots. ِِAlso, the combined inoculation of cyanobacteria and N2-fixing bacteria along with 75% nitrogen led to a significant increase in shoot and root dry weight as well as root yield quality (sucrose and purity). Taken together, these results suggested that the biological treatments were an effective way to improve the salt tolerance of sugar beet as well as the exogenous application of proline. Moreover, the beneficial effect of the cyanobacteria and N2-fixing bacteria on sugarbeet growth, yield and yield quality was attributed to the biologically active substances produced by these microbial strains besides the nitrogen fixation of the diazotrophs, which compensate the reduction of the costly and the environmentally polluted mineral nitrogen fertilizers in the new reclaimed saline-sodic soil.

8/30 IMPACT OF LONG-TERM APPLICATION OF CHEMICAL, BIOLOGICAL AND ORGANIC FERTILIZERS ON SOME SOIL CHARACTERISTICS AND SUGAR BEET PRODUCTIVITY

Mohamed Nour El-Din, M. and Naseer I. Talha

Soil, Water and Environmental Research Institute, ARC, Egypt.

In concrete lysemeter pools of 70cm x70 cm, successive plant crops have been planted since 2001 till 2008 in order to study the accumulation influence of organic matter, compost, (OM); biological inoculation with N2-fixing and phosphate-dissolving bacteria, (BI), bio-organic-fertilization (BF) and organic agriculture (OA) applications compared with conventional chemical fertilization (C) on some soil chemical characteristics and fertility. The study showed the influence of long-term additions on improving soil characteristics and fertility. Sugar beet plants were grown through winter season of 2008/2009 without any additions, in order to know the reflection of the past historical applications on soil fertility. The afore-applied treatments achieved a clear improvement in soil fertility, demonstrated by the increase in the plant root production over that of conventional treatment which ranged from 5.18% for OM to 35% for BF. Root weight/plant, also, increased, ranging from 5% for OM to 34.9% for BI. Regarding NPK-percentages in plant root, the treatments did not exhibit clear consistent effect. On the other hand, the soil analysis demonstrated a remarkable improve in soil chemical characteristics, OM was increased along with increase in N and K percentages, but P% did not show a clear trend. In spite of the absence of significant variation in soil pH, a notable decline in levels of soluble cations and anions as will as EC were recorded due to the long-term application of these amendments in relation to conventional NPK-applied treatment. Therefore, we recommend the continuous application of bio-organic treatment which will lead to the gradual improvement of soil fertility, in addition to production of safe and secure foods away from the excessive use of chemical fertilizers and pesticides, which have harmful effect on man and the environment.

9/30 IMMUNOTHERAPY OF MALIGNANT TUMORS CHALLENGES AND PROSPECTS

Mona F. Mohamed, Ahmed H. Abdel-Aziz, Asmaa N. El-Din and El-sayed M. Aly*

Medical Microbiology and Immunology Department, Oncology Department*,

Sohag Faculty of Medicine, Sohag University.

Background: Immunological treatments are considered now one of the routine methods of cancer therapy including interferon alpha, interleukine-2, monoclonal antibodies and BCG also vaccines against hepatitis and human papilloma viruses prevent certain forms of cancers. Cancer immunotherapy is mostly focused on the generation of large number of activated anti-tumor effector cells by vaccination or adoptive T cell transfer. Aim of the work: this review will provide discussion about anti-tumor immune mechanisms including mechanisms of immune evasion by the tumor and will discuss also aspects of cancer immunotherapy including cancer vaccines and passive immunotherapy approaches. Conclusion: now, immunotherapy is considered as an important method of cancer therapy in combination with conventional methods of cancer therapy like radiotherapy, chemotherapy, and surgery as these methods will augment the results of immunotherapy.

10/30 INFLUENCE OF CYANOBACTERIA AND AZOLLA ON WHEAT CROP CULTIVATED IN A SALINE SOIL

Azza A. M. Abd El-All, Elham M. Aref and Reda M. El- Shahat

Soils, Water and Environ. Res. Inst., Agric. Res. Center, Giza, Egypt

Two field experiments using Azolla pinnata and a mixture of cyanobacteria (Nostoc muscorum and Anabaena oryzae) to evaluate their role on growth and yield of wheat grown in saline soil at South El- Hossinia Research Farm Station, El- Sharkia Governorate, Agric. Res. Center, (ARC). Egypt, during two successive winter seasons 2009/2010 and 2010/2011 were designed. Influence of Azolla (fresh and dry) and cyanobacteria suspension as seed soaking ,filtrate for spray treatments and urea (75kg-N) as nitrogen fertilizer on growth, yield and yield components of wheat (Triticum aestivum cv.sakha 69) as well as their effect on some chemical properties and biological activities was studied. Results revealed that high organic matter was due to the use 25kg-N-urea +25kgN dry Azolla+17L cyanobacteria (T11) was recovered. All the tested biofertilizer treatments decreased both EC and pH compared to the control treatment. Results also exhibited a favorite influence of foliar application for both Azolla and cyanobacteria (T11), since it gave significantly higher wheat yield, yield components and N P K contents of both grains and straw compared to the other treatments and control . The treatment (T11) of 25kg-urea+25kg N- Azolla+17L cyano. which applied as foliar spray gave also the high values of soil biological activity parameters. The obtained results revealed beneficial aim to use cyanobacteria and Azolla (dry or fresh) applied as foliar +25kgN-urea to wheat plants exhibited an economical view that it can save about 50% of the mineral nitrogen amounts and could be used as a good tool to promote wheat productivity and yield quality as it provides an eco-friendly biological approach under saline soil conditions.


11/30 BENEFICIAL EFFECTS OF AZOLLA AND CYANOBACTERIA ON RICE PRODUCTION IN SALINE SOIL

Elham, M. Aref, Reda M. El- Shahat and Manal A.H. EL-Gamal

Agricultural Microbiology Dept., Soils, Water & Environment Res. Inst. (SWERI), Agric. Res. Center(ARC), Giza, Egypt

Two field experiments were carried out at Sahl El-Hossinia Research Station during two successive growing seasons of (2008-2009) to evaluate the influence of Azolla and Cyanobacteria as biofertilizer in combination with urea (46.5 % N) as nitrogen fertilizer on rice productivity cultivated under saline soil conditions. Also to study their effect on some soil characteristics (chemical properties and biological activity). Where, Azolla was applied either fresh or dry as soil application or as extract for seed soaking and foliar spray. While, Cyanobacteria was used as extract for seed soaking and foliar spray only. Both of Azolla and Cyanobacteria were applied alone or in combination comparing with control treatment (full dose of NPK). Results revealed that, application of Azolla and Cyanobacteria spraying, increased significantly plant height, number of tillers/hill, plant dry weight, grains yield and weight of 1000 grain. On the other hand, straw yield decreased with application of Azolla and Cyanobacteria. Azolla and/or Cyanobacteria significantly increased nitrogen, phosphorus and potassium content of rice grains and straw as compared with control treatment. Inoculation of rice with Azolla and/or Cyanobacteria under saline soil condition with full dose of nitrogen, showed positive significant response on soil biological activity. It increased the count of bacteria, Azospirilla and Cyanobacteria. However, the effect of Azolla either applied alone or mixed with Cyanobacteria, resulted in an increase in CO2 evolution, DHA and N2-ase activity over control. Utilization of Azolla as a tool for increasing soil fertility, either alone or in combined with Cyanobacteria, increased the soil O.M. and nitrogen content. However, incorporation of Azolla fresh or dry decreased the soil pH and soil salinity as compared with control. Currently, it could be concluded that the conjunctive use of Azolla along with Cyanobacteria resulted not only in highest and sustainable rice crop yield but also enhanced soil status.

12/30 STUDIES ON BACTERIAL ENDOPHYTES COLONIZATION IN VITRO AND IN VIVO TO UNDERSTAND ITS ROLE IN PLANT–MICROBE INTERACTIONS

Eman, A. Tantawy

Agriculture Microbiology Department, Soils, Water and Environment Research Institute, Agriculture Research Center, Giza, Egypt

To study the interaction between plant and endophytic bacteria, an experiment in vitro was carried out through wheat plants inoculation with endophytic isolates (Klebsiella pneumoniae and Azospirillum brasilense) and cyanobacteria association (mixed strains of Nostoc muscorum, Nostoc calcicola, Anabaena oryzae and Clyndrospermum muscicola)for comparison. These isolates were inoculated in wheat plantsusing sterilized gnotobiotical system under a growth-chamber conditions to study the ability of endophytic bacteria to re-entry into the root tissues and their distinctive behavior as endophytes. The same isolates used to perform an experiment in vitro, conducted at the Experimental Farm of Agricultural Research Center, Ismailia Governorate, Egypt, to detect the response of wheat growth and productivity to these isolates. In vitro experiment, the anatomical studies exhibited that Klebsiella pneumoniae was showed with excess colonies intracellular of root sections over A. brasilense, which had exhibited less colonies. Meanwhile, cyanobacteria had no colonies shown intracellularly as well as the control. Results emphasized that the endophytic strains have the ability to entry the plant tissues with a large number showed preceding in most of the tested plant parameters studied (straw yield, grain yield and NPK uptake by straw and grains). That means the endophytic bacterial efficiency in plant entry resulted in its high ability to produce their acts from nitrogen fixation, phosphate and potassium solubilization, produce PGP substances like IAA, EPS and sidrophores until forming antifungal and bio-control activity.

13/30 LINCOMYCIN B BIOSYNTHESIS BY STREPTOMYCES RAMULOSUS, A-MM-24: FERMENTATION, PURIFICATION AND STRUCTURAL ELUCIDATION

Nagwa Mahmoud Sidkey, Maha Abd Al-Rahman Abo-Shadi*

and Abeer Mohammad Al-Mutrafy**

Botany & Microbiology Department, Faculty of Science (Girls), Al-Azhar University, Cairo, Egypt

*Microbiology and Immunology Department, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt

**Biology Department, Faculty of Science (Girls), Taibah University, KSA

Effort has been made in this study to improve antimicrobial agent production by the strain Streptomyces ramulosus, A-MM-24, isolated from Al-Madinah Al-Munawwarah's soil-KSA, through optimization of some nutritional and cultural conditions. A trial was also done to make purification and characterization of the antibiotic obtained from this local strain. Its biological activity to control two multi-resistant human pathogenic bacteria (E.coli and methicillin resistant Staphylococcus aureus, MRSA) was also studied. The optimum production of the antimicrobial agents by S. ramulosus, A-MM-24 was achieved in starch nitrate medium supplemented with starch & peptone as carbon and nitrogen sources, respectively, with further process parameters such as incubation temperature at 30˚C, initial pH 7 of the medium, inoculum size 33.75x105 spore/ml and incubation period of sixteen days under shaking. Production of the antibiotic was done using batch-scale fermentation under all the previously mentioned optimum conditions. The active metabolite was extracted using ethyl acetate (1:1, v/v) at pH 7.0. The separation of the active ingredient and its purification was performed using column chromatography technique. Elemental and spectroscopic analysis was used for the structure elucidation of the antibiotic produced. The physico-chemical characteristics of the purified antimicrobial agent have been also investigated. This antibiotic could be suggested of being belonged to Lincosamide group (Lincomycin B antibiotic) with a suggested empirical formula of C17 H32 O4 N2 S.

14/30 BIOCONTROL AGENTS AS A TOOL FOR REDUCING CHEMICAL NEMATICIDE USING FOR CONTROL MELOIDOGYNE JAVANICA INFECTED PEANUT PLANT

Eman A. Tantawy and Hanaa S. Zawam*

Soil water and Environ. Res. Inst., Agric. Res. Center, Giza (Egypt)

*Plant pathology Res. Inst., Agric. Res. Center, Giza (Egypt)

To examine the bio-control agents against Root-knot nematodes(Meloidogyne javanica) which infect peanut plants, two pre-experiments were carried out under Laboratory and greenhouse conditions, Bradyrhizobium sp., Serratia marcescens, Saccharomyces cerevisiae, and Glomus macrocarpum were used as biocontrol agents besides Bacillus subtilis as biopesticides product and furadan as a chemical nematicide. The successful treatments were chosen due to their effectiveness on percentage reduction in nematode populations, maximize plant production and caring safety environmental by reduction nematicides using. The chosen treatments applied in soil naturally infected with nematode in the experimental field in Nubaria, (Behira Governorate) north Egypt. Results exhibited significantly reduction in all nematode developmental stages in plant and soil, ex. number of egg masses, galls and nematodes in 250 cm3 soil with all bio-control agents consequence plant parameters and grain yield increase significantly and the priority was for activator yeasts with furadan 50 % and/or bradyrhizobia in grain yield. In decreasing of nematode parameters the priority recorded with cleanroot 100% and furadan100% respectively. It have been noticed that the biocontrol agents more effective when using combined with each other than when used solely.

 

15/30 EFFECT OF CYANO- AND ENDO-BACTERIA ON WHEAT PRODUCTIVITY UNDER NITROGEN LIMITATION OR IRRIGATION REGIME IN CALCAREOUS SOIL

Soha S.M. Mostafa, Eman A. Tantawy, Manal A.H. El Gamal and

Abd El-Hady Kh. Abd El-Halim

Soils, Water and Environ. Res. Inst., Agric. Res. Center (ARC), Giza, Egypt.

Two field experiments were conducted on calcareous soil of Nubaria Agric. Res. Station, Agric. Res. Centre (ARC)-Egypt, on wheat crop. The first experiment was carried out to study the effect of cyanobacteria and endophytic bacteria inoculations either solely or in mixture on wheat productivity with half dose of nitrogen fertilization. Results revealed that soil microbial activity and soil microbial density of wheat rhizosphere, after 75 days from wheat planting and after harvest were affected by the inoculation with cyanobacteria and endophytic bacteria strains either sole or in combinations. N2-fixing cyanobacterium Anabaena oryzae solely achieved the highest significant records of soil microbial activity and soil microbial density. On the other side, some strains couldn't individually improve soil microbial parameters, but their performance positively improved when combined in mixtures. Results emphasized on the collaborative relationship between cyanobacteria and endophytic bacteria. The inoculation increased all the biological activities in plant rhizosphere, dehydrogenase and nitrogenase enzymes as well as total bacterial, total cyanobacterial and N2-fixers bacterial counts. The inoculation increased consequence biological and grain yield in addition to the content of NPK in plant and grain in the same time with increased of N-NO3 and N-NH4 in soil after harvest. While the second experiment was carried out to detect the influence of the mixed inoculation of cyanobacteria and/or endophytic bacteria on wheat production under different irrigation levels (100% of ETc, 75% of ETc and 50% of ETc). Results indicated that the biological treatments helped to save about 25% of irrigated water in calcareous soil and did not affect the productivity of wheat as compared with the control. The interactive effect between cyanobacteria and endophtic bacteria achieved the best results on the productivity of wheat and decrease the applied amount of Irrigation water.

 

16/30 COMPARISON OF MRSASelect, ORSAB AND BBL CHROMagar MRSA IN THE SURVEILLANCE FOR METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS

Eman Ahmed El-Seidi and Mona Gamal El Din Nada

Medical Microbiology & Immunology Department, Faculty of Medicine,

Cairo University, Cairo, Egypt

Methicillin-resistant Staphylococcus aureus (MRSA) are responsible for nosocomial and community-acquired infections. Rapid laboratory diagnosis is critical for its treatment, management and prevention. Evaluation and comparison of the potential for MRSA detection using three commercially available chromogenic media, namely MRSASelect (Bio-Rad), ORSAB (Oxoid) and BBL CHROMagar MRSA (BD Diagnostics) was performed. The study was done on a total of 753 pre-enriched nasal swabs, cultured on chromogenic media and mannitol salt agar. MRSA was detected and confirmed by standard phenotypic and genotypic tests. The study revealed 138 MRSA strains that could be confirmedphenotypically and genotypically. Two confirmed MRSA strains failed to be detected by any of the three tested chromogenic media. The three chromogenic media showed sensitivities over 82% and specificities over 89%, however, MRSASelect gave marginal more accurate results for MRSA identification. Chromogenic media were cheap accurate screening tools for MRSA, particularly suitable for limited resource settings.

17/30 IMPACT OF HELICOBACTER PYLORI INFECTION IN PATIENTS UNDERGOING GIT ENDOSCOPY SUSPECTED FOR CANCER

Hanaa M. Alam El-Din, Abdel Gawad M. Hashem*, Yasser M. Ragab*, Abeer M. El Sayed** and El-Chaimaa B. Mohamed

Department of Cancer Biology, National Cancer Institute.

*Department of Microbiology and Immunology, Faculty of Pharmacy,

Cairo University.

** Department of Pathology, National Cancer Institute, Cairo University.

            Helicobacter pylori (H. pylori) is a gram negative flagellated microorganism affecting half of the world's population. It is considered as a major cause of chronic gastritis, peptic ulcer disease, gastric carcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. The aim of this study was to investigate the prevalence of H. pylori and its clinical impact in patients undergoing GIT endoscopy suspected for cancer and to evaluate and compare two different non invasive methods for diagnosis. This study included 47 patients undergoing GIT endoscopy (12 patients with gastritis, 8 with gastric adenocarcinoma, 12 with MALT lymphoma, 15 with other cancer types) as well as 3 patients who were disease free so added to the control group. The study also included a control group of 20 apparently healthy subjects. All patients and control group were subjected to serological testing for H. pylori IgG antibody by ELISA and H. pylori stool antigen by stool antigen test (HpSA). The total positive cases for H. pylori by stool Ag test were 54/67 (80.6%) and by serum IgG Ab test 63/67 (94%) with a sensitivity of 84.1%, specificity of 75% (p= 0.02). Thirty four patients out of 47 (72.3%) were positive by both the H. pylori stool Ag (HpSA) and serum Ab tests, while 2 (4.25%) were negative by both tests. However in the control group, 19 patients (82.6%) were positive while only one patient (4.34%) was negative by the two methods. Stool Ag (HpSA) test for H. pylori infection was positive in 93.3% among the different cancer types: it varied from 62.5%, 75%, 77.2%, 77.8% to 81.8% in MALT lymphoma patients, gastric adenocarcinoma patients, gastritis patients and other cancer type patients, respectively. H. pylori infection was higher in rural (81.8%) than in urban patients (77.2%) (p= 0.764), and in males (85.7%) than in females (72.2%) (p= 0.304). The old age group (61-80 years) was at higher risk of H. pylori infection (90.9%) than the young (20-40 years) and middle age groups (41-60 years) (87.5% and 72%, respectively) (p=0.323). We noticed a high incidence of H. pylori infection in cancer patients and it was associated with MALT lymphoma and adenocarcinoma diseases. Early detection and treatment of H. pylori could decrease and may prevent the development of gastric cancer and MALT lymphoma diseases. Serum antibody test was sensitive but not sufficiently specific, so it could not be used alone for the detection of H. pylori, but is better used for screening of infection followed by another confirmatory test.

    

18/30 CONVENTIONAL AND MOLECULAR METHODS FOR DIAGNOSIS OF BACTERIAL MENINGITIS IN EGYPTIAN AND SAUDIAN PEDIATRIC PATIENTS

Marwa A. Mansour,Omar M. Eldaly and Asif Jiman Fatani*

Medical Microbiology and Immunology Department, Faculty of Medicine, Zagazig University, Egypt.

*Medical Microbiology and Immunology Department, Faculty of Medicine, King Abd El-Aziz University, Saudi Arabia.

Diagnosis of bacterial meningitis (BM) remains a challenge to the clinician because of its rapid lethal course lacking the consistency to particular clinical signs and symptoms. Moreover, in many clinical settings use of short course antibiotic therapy prior to lumbar puncture reduces the chance of isolation of bacteria in CSF culture making the diagnosis difficult. The present study was done to compare between the conventional and molecular methods for identification of organisms causing BM.Cerebrospinal fluid (CSF) samples were collected from 110 suspected BM patients; 60 patients were admitted to Zagazig University Children's Hospital, Egypt and 50 patients were admitted to King Abd El-Aziz University Hospital, Saudi Arabia. Those patients developed signs or symptoms suggestive of BM. Their ages ranged from 12 days to 13 years (mean±S.D. 35.35±27.62 months). The samples were subjected to physical, chemical and cytological examinations. Conventional methods of bacterial identification were direct microscopic examination, culture and biochemical reactions. On the basis of findings of culture, Gram staining and WBC counts in CSF, patients were divided into 3 groups; infection, probable infection and no evidence of infection groups. Bacterial broad range- polymerase chain reaction (BBR-PCR)/ restriction fragment length polymorphism (RFLP) was the used molecular method.This study showed a significant difference (P< 0.001) in all parameters except the age in infected group followed by probable infection group in comparison with group of no evidence of infection.The sensitivity, specificity, positive predictive value and negative predictive value of BBR-PCR were 89.4 %, 87.3 %, 84.0 % and 91.7 % respectively with a statistically significant difference (P < 0.001) in comparison with culture. In Egypt, Escherichia coli (57.14 %), Haemophilus influenzae type b (52.63 %) and Streptococcus pneumoniae (100 %) were the bacteria with highest isolation rate in neonates, infants and young children and older children respectively while in Saudi Arabia, they were E.coli (50 %), S. pneumoniae (57.14 %) and S. pneumoniae (66.67 %) in the same age groups. On comparing the detected bacteria by BBR-PCR/RFLP in Egypt and Saudi Arabia, only H. influenzae type b show higher isolation rate (34.48 % in Egypt versus 9.52 % in Saudi Arabia) with a statistically significant difference (P=0.041).The results suggested that BBR-PCR/RFLP can be used as a valuable supplementary test in routine clinical practice for diagnosis of BM particularly in hospital setting.

19/30 MEASURING THE ANGIOGENIC MARKERS IN VIRAL HEPATOCELLULAR CARCINOMA AND CHRONIC VIRAL HEPATITIS PATIENTS

Abeer A. Ghazal, Mohamed A. Abdel Mohsen*, Neveen A.Hussein*

and Mohamed Farouk**

Departments of Microbiology, Applied Medical Chemistry, Medical Research Institute*, and Clinical Oncology and Nuclear Medicine Department**, Faculty of Medicine Alexandria University.

The occurrence of hepatic angiogenesis has been described in liver conditions such as viral hepatitis, cirrhosis, and hepatocellular carcinoma(HCC). Observations suggest that tumor angiogenesis is linked to a switch in the equilibrium between positive and negative regulators. Vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), angioststin (AS) and endostatin (ES) have been reported to be related with angiogenesis and can be detected in both tissue and blood. The aim of this study was to investigate the serum level of VEGF, Ang-2, AS and ES in Hepatitis C virus (HCV), Hepatitis B virus (HBV) infected and HCC patients to study the possibility of their use as prognostic non invasive markers. The study included 120 subjects divided into 6 groups. Group I, included 20 chronic HCV infected patients documented by the presence of HCV antibodies and HCV RNA in the serum. Group II, included 20 anti HCV positive patients but serum HCV RNA negative. Group III included 20 HBsAg positive HBVDNA positive patients. Group IV included 20 HBsAg positive HBVDNA negative patients. Group V included 20 clinically diagnosed HCC patients associated with HCV infection and group VI included 20 healthy individuals as control. Serum levels of VEGF, Ang-2, AS and ES were measured by ELISA technique. In HCV patients, the levels of sAng-2 and sAS were significantly elevated in group I and group II when compared to that of group VI, while the level of sVEGF in Group I and Group II did not differ from that of Group VI. In HBV patients, the levels of sVEGF, sAngiop-1, sES and sAS in group III were significantly higher than group VI. In group III the levels of all angiogenic markers except sAS were significantly higher than those of group I and group IV. In HCC patients, a significant increase in the serum levels of all angiogenic markers was noticed when compared to all groups. So, we can conclude that measurement of serum levels of both (proangiogenic) and (anti-angiogenic) factors could be useful as noninvasive markers of disease progression in HCV and HBV patients.

20/30 EFFECT OF OZONE TREATMENTS ON FUNGAL GROWTH AND DETOXIFICATION OF CORN

Mona M. Abd El-Magied, Salah H. Abou Raya, Mona. M. Abdelgalil*

and May M. Amer*

Food Sci. & Technol. Dept., Fac. Agric., Cairo Univ., Giza, Egypt.

*Food Toxicology & Contaminants Department, National Research Center, Dokki, Cairo, Egypt.

Mycotoxicoses are diseases caused by consumption of diets contaminated with mycotoxins, a special class of fungal secondary metabolites. Aflatoxins and fumonisin, the main toxins synthesized by toxigenic stocks of Fusarium spp. and Aspergillus spp., respectively, can coexist in corn grains. Samples of collected corn from 3 different Governorates in Egypt had different percentage of infection in the range of 4.9 to 6.3% and from 5.4 to 6.2% for white and yellow corn grains, respectively. Total fungal count (TFC) which isolated from white corn grain samples yielded 358 fungal isolates equal 50.78 %, while the (TFC) isolated from yellow corn grain samples resulted 347 fungal isolates equal 49.22 %. Five fungal genera were identified as a-contaminant of corn grain samples in this work. These are Aspergillus, Fusarium, Mucor, Penicillium and Rhizopus. Aspergillus genus was the most dominant of (TFC) followed by Fusarium genus, while Penicillium was less fungal frequency occurred. Ozone is an effective for mycotoxin destruction and microbial inactivation ,while has a minimal or no effect on corn grain quality. Application of ozone was found to decrease (TFC) in all the tested samples. The production of aflatoxin from A. flavus reached 1258.1 µg/kg and the production of fumonisin from F. moniiforme reached 213.25 mg/kg, in the contaminated corn. Using ozone 40 mg/L for 30, 60 or 180 min. led to gradual decrease in toxin production. A significant decrease in body weight of all the groups of rats fed with contaminated aflatoxin or fumonisin was recorded. Ozonation treatment affected positively on the contaminated white corn diets that increased the body weight for all the groups. The corn contaminated with those mycotoxins caused significant increases in liver enzymes (ALT and AST) as well as lipid parameters in rats besides deterioration for kidney function which was evident by the increase of kidney parameters such as (creatinine, urea, and uric acid).

21/30 MICROBIAL INOCULATION IMPACT ON YIELD PRODUCTIVITY OF CLOVER IN HEAVY TEXTURED SOIL

Heba Sh. Shehata, Elham M. Aref, Heba M.A. Khalil

Microbiology Dept. soils, water and Environment Inst. Agaric. Res. Center,

Giza, Egypt.

Two field trails were executed in winter seasons of 2008-2009and 2009-2010 to evaluate potentiality of some rhizobacterria to support nodulation, growth and productivity of clover barseem in clay soil. Results of 5 cuts indicated that inoculation with Rhizobiumleguminosarum together with Bacillus subtilis and B. polymyxa induced positive effects on dry yield (30.6 % increase over control) and fresh yield (27.33%). Single inoculation of clover seeds with rhizobium significantly increased number of tillers, dry weight of roots, leafiness % and some chemical constituents i.e crude protein, crude fiber, ash and nitrogen-free extract. When plants wereinoculated with rhizobium combined with either of B. subtilis or B. polymyxa or mixture of both induced positive effects on rhizobium nodulation on plant roots. In vitro examination revealed that all tested rhizobacteria exhibited positive impressionfor phosphatesolubilization, Indol Acetic Acid (IAA), Gibberellic Acid (GA) excration,beside siderophores and hydrogen cyanide production. Multifunctional PGPR conspicuously promoted nodulation and yield of clover in heavy textured soil. The improvement of nodulation pattern was strengthen in dual inoculation with rhizobium in combination with any of PGPR strains indicating promotive effect on nodules formation on the root system. This promotive effect may be occurred by providing the clover some synergistic substances such as auxinsan fluorides. Combinations of Bacillus subtilis, B. polymxya and Bradyrhizobium with half dose of chemical fertilizers showed a significant increase shoot weight, root length,nodules and root weight were graeter for the combination with half dose of chemical fertilizers compared to the control.

22/30 VIRAL LOAD AND OXIDATIVE STRESS IN EGYPTIAN CHRONIC HEPATITIS C PATIENTS

Bassel Mamdouh Hussein, Eman Ahmed El-Seidi* and Youssef Abdallah Abou-Hamed*

Central Labs, Ministry of Health, *Department of Medical Microbiology & Immunology, Faculty of Medicine, Cairo University

Hepatitis C virus (HCV) is major cause of viral hepatitis in Egypt. This viral infection frequently does not resolve and chronic HCV carriers may then progress to serious liver diseases.The overproduction of reactive species results in oxidative stress, causing an imbalance in the host. HCV infection is associated with severe alterations of the host redox status, thus playing important roles in the development of HCV-associated liver diseases. Determination of the presence of any association between chronic HCV infection and the oxidative stress, namely correlating between the HCV viral load and liver enzymes, malondialdehyde (MDA) and catalase (CAT) enzyme in chronic HCV patients was done. Evaluations were done on a total of 150 chronic HCV Egyptian patients, having variable viral loads, in addition to a control group of 50 healthy subjects. HCV antibodies were detected by ELISA and HCV viral load was determined by Real Time PCR. Chemical detection of serum alanine transaminase (ALT), aspartate transaminase (AST), malondialdehyde (MDA) and catalase enzyme was performed. The study revealed that a statistically significant correlation between the liver enzymes and oxidative stress markers has been detected. Moreover, a statistically significant positive correlation existed between MDA and the HCV viral load and a statistically significant negative correlation existed between the catalase enzyme and HCV viral load. Oxidative stress is a significant feature of chronic HCV infection. The higher the viral load, the more prominent the oxidative stress is.

23/30 THE PREVELANCE OF HBEAG AMONG HBSAG POSITIVE BLOOD DONORS IN MINIA GOVERNORATE.

Mohamed Abdel-Hamid Ahmed, Amgad Abo Ouf, Sahar Mohamed Abo El Euoon, Rasha Mohamed Mahmod and Nagwa Nabel Zedan*

Department of Microbiology and Immunology, Faculty of Medicine, Minia University and **The Regional Blood Transfusion Center, Minia, Eygpt.

Background: Hepatitis B virus is now recognized as one of the major health problems allover the world. Blood is the important vehicle for its transmission. It was reported that 15%-40% of HBV infected patients would develop cirrhosis, liver failure, or hepatocellular carcinoma. Objective: The study aims to determine the prevalence of HBeAg among HBsAg positive blood donors in blood Transfusion Services Center in Minia Governorate. The study aims also for the detection of viral DNA by PCR in HBeAg seropositive donors, as a marker for replication. Methods: The study was done on 15.500 volunteers blood donor attending the Regional Blood Transfusion Center in Minia and were subjected to blood screening for HBsAg by enzyme immunoassay (EIA) technique .Two hundreds blood samples were selected randomly from HBsAg seropositive and were subjected to detection of HBeAg by (EIA) technique and detection of viral DNA by PCR in HBeAg seropositive. Results: Among 15.500 blood samples, 307 (1.9%) were HBsAg positive. From the 200 selected samples, 21(10.5%) were HBeAg positive and all of them were HBV DNA positive.

24/30 DIVERSITY AND PHYLOGENETIC ANALYSIS OF LuxS and RsbA GENES IN BIOFILM FORMING PROTEUS MIRABILIS

Reham Wasfi; Amro S. Hanora*; Lamiaa E. Mansour**; Ola A. Abd El-Rahman***; Abd El-Gawad M. Hashem** and Mohamed S. Ashour***

Microbiology Dept. Fac. Pharmacy, October University for Modern Sciences and Arts, Giza, Egypt

* Microbiology Dept. Fac. Pharmacy, Suez Canal University, Ismailia, Egypt

** Microbiology Dept. Fac. Pharmacy, Cairo University, Cairo, Egypt

*** Microbiology Dept. Fac. Pharmacy, El-Azhar, Cairo, Egypt

Quorum sensing phenomenon is thought to be involved in organizing many of the mechanisms that enable the bacteria to survive under unfavourable conditions. One of these mechanisms is biofilm formation which is responsible for chronic infections and antimicrobial resistance. Using molecular tools for studying quorum sensing is more preferable than other methods due to specificity and accuracy. Primers were designed for ampilification of LuxS, autoinducer 2 synthetizer, and RsbA , part of the two component regulatory system, genes. PCR products were sequenced and then phylogenetically analyzed to give hypothesis on the possible function of these genes and its possible role in biofilm formation. The designed primers were successful in ampilifying the targeted gene. The phylogenetic analysis of the ampilified genes showed similarity to homologues genes in enterobacteriaceae, while the phylogenetic analysis of the 16S rRNA gene showed similarity of different isolates to Proteus mirabilis strains from different origins. The homology of the ampilified LuxS and RsbA genes from Proteus mirabilis isolates to homologues genes in some enterobacteriaceae indicated that these genes may have role in bioflm formation, swarming, toxin production and antibiotic production.

25/30 TUMOR NECROSIS FACTOR ALPHA AS A NOVEL PROGNOSTIC MARKER IN VIPERIDS ENVENOMATION AND THE POSSIBLE THERAPEUTIC ROLE OF ITS ANTIBODIES VERSUS ANTIVENOM

Nehad M. Sayed, Assem A. El Habashy*, Nahla H. Tolba*, Yasser F. Abd El Monem*, SohaK. Seif Elnasr* and Suzi S. Atallah**

Microbiology and Immunology Department,*Forensic Medicine andClinical Toxicology Department and **Histology Department, Faculty of Medicine,

Ain Shams University

Viper envenomation is one of the common toxicity accidents characterized by prominent local tissue damage that may lead to permanent tissue loss. TNFα at the site of the bite contribute in the pathogenesis of local injury, and antivenom is ineffective in suppressing the local reaction induced by viper venom. This study comprised clinical and experimental parts. The aim of the clinical part was to measure TNFα in the sera of patients bitten by viperids and correlating its level with severity of the cases.While the aim of the experimental part was to assess the effects of anti-TNFα antibodies and the antivenom alone and in combination on the local and the systemic effects of viper venom in experimental animals. The clinical part of this study included 20 symptomatic patients admitted to the Poison Control Center (PCC) Ain Shams University because of viper bites,in addition to ten healthy controls. Every included subject was subjected to thorough history taking and clinical examination. CBC, coagulation profile, renal profile, CPK level and serum TNFα level were measured twice for each patient, first on admission then after 12 hours. The experimental study was carried out on 100 adult albino rats (180-220 grams). They were divided into 5 groups, each group comprised 20 rats. Group I was the negative control group, group II was given viper venom where groups III, IV and V the effects of the venom, the antivenom and anti-TNFα antibodies were studied. The clinical part of this study showed that on admission tumor necrosis factor alpha (TNFα) level is increased in sera of patients envenomated by viper bites. This high level was correlated with decrease in RBCs count, hemoglobin concentration, platelets count, fibrinogen level and the increase in WBCs count, PT, PTT, FDPs level and CPK level. After 12 hours,the level of TNFα is decreased but is still higher than controls, a significant correlation was found between its level and the same previous parameters. The experimental part of the present study showed that administration anti-TNFα antibodies protected the tissues from the local devastating effects of the viper venom. Normalization of the systemic alterations were achieved with the administration of antivenom either alone or combined with anti-TNFα antibodies. TNFα level is increased in cases of viper envenomation and could be used as a good marker for prognosis in these cases. In addition, the experimental part of the present study showed that the best treatment for local and systemic alterations was obtained with the combined action of the antivenom and anti-TNFα antibodies.


26/30 DETECTION OF CHLAMYDIA TROCHOMATIS, UREAPLASMA UREALYTICUM AND NEISSERIA GONORRHOEAE ENDOCERVICAL COLONIZATION IN WOMEN WITH UNEXPLAINED INFERTILITY BY MULTIPLEX POLYMERASE CHAIN REACTION

Enas K. Abo Elmagd, Naglaa M. Moharam* and Wafaa A. M.hamed**

Microbiology, *Obstetrics & Gynecology and **Dermatology & Venereology Departments, Faculty of Medicine for Girls, Al- Azhar University, Cairo.

Genital infection represents one of the most important causes of infertility, affecting: fallopian tubes, endometrial mucosa and sperm parameters. This study was aimed to assess the incidence of Chlamydia trachomatis, Ureaplasma urealyticum and Neisseria gonorrhoeae endocervical colonization in women with unexplained infertility compared to fertile women by multiplex polymerase chain reaction (M.PCR). Endocervical swabs were taken from 50 women with unexplained infertility {(27 (54.0%) with primary infertility and 23 (46.0%) with secondary infertility} their ages ranged from 16 - 40 years and from 50 fertile women with age range from 19 - 40 years as a control group. C. trachomatis, U. urealyticum and N. gonorrhoeae were detected in the endocervical specimens by M.PCR. Chlamydia trachomatis was detected in 14/50 (28.0%) of the infertile group compared to 2/50 (4.0%)ofthecontrol group with highly significant difference (P<0.0001). Ureaplasma urealyticum was detected in 18/50 (36.0%) of the infertile group and in 5/50 (10%) of the control group with highly significant difference (P<0.0001). Six out of fifty (12%) of the infertile cases had concomitant colonization with both C. trachomatis and U. urealyticum. Only one infertile case was positive for N. gonorrhoeae while all fertile women were negative. C. trachomatis and U. urealyticum colonization rates were higher in secondary infertility rather than in primary infertility, {(34.8% and 47.8%) versus (22.2% and 25.9%)} respectively. In conclusion: A significantly high colonization rates of C. trachomatis and U. urealyticum were detected in infertile women much more often in secondary infertility cases. Screening for cervical colonization with these sexually transmitted organisms in a single test by M. PCR may be a promising method for detection of asymptomatic pelvic infection in patients with infertility.

27/30 A TWO-YEAR (2006-2008) SURVEILLANCE OF ANTIMICROBIAL RESISTANCE AMONG NOSOCOMIAL PATHOGENS ISOLATED FROM NATIONAL HEART INSTITUTE, CAIRO, EGYPT

Aza R. Mohamed, Noha G. Khalaf* and Ramadan A. Al-Domany**

National Heart Institute, Cairo, Egypt; *Department of Microbiology and Immunology, Faculty of Pharmacy, Modern Science and Arts University, Egypt; **Department of Microbiology and Immunology, Faculty of Pharmacy, Helwan University, Egypt

High resistance rates to extended spectrum cephalosporins (ESC) among multidrug resistant (MDR) Gram negative (GN) isolates as well as methicillin resistant S. aureus (MRSA) have been reported in Egypt. In this study, the prevalence of MDR GN and Gram positive (GP) isolates at the National Heart Institute (NHI), Cairo, Egypt during 2006-2008 was reported. During a 2-year period, 300 nosocomial isolates were obtained from hospitalized cardiac patients in ICU and non-ICU wards at the NHI, Cairo, Egypt. Isolate identification at the species level and MIC determination were performed using MicroScan panels (Dade-Behring, USA). Genotyping of K. pneumoniae (KP) isolates was carried out using Enterobacterial Repetitive Intergenic Consesus sequence PCR assay. Out of the 197 GN rods, KP were (40%), followed by Enterobacter spp. (9.3%), and E.coli (5%). Other GN isolates included P. aeruginosa and A. baumannii. Among the 103 GP cocci, S. aureus comprised 13.3% and 19.3% were coagulase negative staphylococci. KP showed a high resistance rate (77%) to ESC, and 44% were resistant to cefoxitin (FOX). Resistance to ciprofloxacin (CIP) was 75% and 84% were resistant to gentamicin (GEN). However, 96% were susceptible to imipenem (IPM). Cross-resistance to ESC and CIP was detected in 59 out of 80 KP isolates (73%). Among the 35 FOX resistant isolates, 30 were resistant to ESC. For SA, 72% were resistant to oxacillin (MRSA), 70% to GEN, and 50% to clindamycin. Susceptibility to CIP was 63%, but 100% remained susceptible to vancomycin (VAN). A common genotype of the K. pneumoniae isolates was detected among 7 patients and a hospital staff. The prevalence of MDR isolates was high at the institute. IPM was effective against KP isolates and VAN was the treatment of choice for MRSA isolates. The prevalence of KP isolates was presumably caused by a single strain circulating between SICU patients and hospital staff.

28/30 ASSESSMENT OF   THE FREQUENCY OF NATURALLY OCCURRING REGULATORY T CELLS IN SYSTEMIC LUPUS ERYTHMATOSUS PATIENTS

Kawther Ibrahim and Sahar Eladawy*

Microbiology and *Internal Medicine Departments, Faculty of Medicine, Ain Shams and *Al Azhar University

Systemic lupus erythmatosus (SLE) is characterized by the presence of autoantibodies associated with a dysregulated B-cell response to multiple antigenic specificities. The presence of elevated titers of autoantibodies is responsible for many clinical and pathologicmanifestations of the disease, since autoantibodies can bind to self antigens, remain entrapped in tissue, and promote localinflammation that can eventually lead to tissue damage and loss of organ function CD4+CD25highFoxp3+ regulatory T cells (Tregs) suppress the proliferation and release of cytokines in several subsets of immune cells. By doing so and by maintaining immune tolerance in peripheral tissues, Tregs contribute to avert autoimmunity. The aim of this work is to asses the frequency of CD4+CD25highFOXP3+ T cells (Tregs) in active SLE patients and correlates them with disease activity and remission of symptoms after treatment in comparison to healthy controls. Fifteen patients with SLE underwent, history taking, clinical examination, laboratory investigation as blood picture, liver function, kidney function, Anti DNA, C3, flowcytometric analysis to assess Tregs. Tregs are significantly decreased in active SLE patients mean0.4713±0.2393 versus 1.218±0.2703 in control. However a significant increase in these cells occurred after corticosteroids treatment mean 0.6580±0.2234 versus 1.218±0.2703 in control.

29/30 THE SIGNIFICANCE OF URINARY SOLUBLE VCAM-1 AND ICAM-1 AS MARKERS FOR DISEASE ACTIVITY IN SYSTEMIC LUPUS ERYTHEMATOSUS PATIENTS

Lamiaa A. Adel, Sherin M. Hosney*, Makram F. Attalah and Dahlia Abdel Mohsen*

Departments of Microbiology and Immunology, *Internal Medicine and Rheumatology, Faculty of Medicine, Ain Shams University

Background: intercellular adhesion molecules -1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are members of the immunoglobulin superfamily that are expressed in normal kidney. Their expression is up-regulated in the renal tissue of patients with lupus nephritis (LN). Objective: to determine the urinary level of   soluble VCAM-1 and ICAM-1 in SLE patients and correlate their level with disease activity & lupus nephritis. Subjects & Methods: The present studyincluded 30 patients with SLE who fulfilled at least four of the American College of Rheumatology (ACR) revised criteria for SLE representing group I, this group was further subdivided into two groups according to the presence of lupus nephritis.Group II included 15 healthy age and sex matched individuals as controls. 24 hours urine samples were collected for the determination of soluble VCAM-1 and ICAM-1 levels by enzyme linked immunosorbent assay (ELISA). Disease activity was assessed clinically by SLAM score. Results: the urinary soluble VCAM-1 and ICAM-1 levels were significantly elevated in SLE patients compared to controls (p<0.0001). Moreover, their levels were found to be higher in patients with lupus nephritis when compared with patients without lupus nephritis p= 0.004 and 0.007 respectively. VCAM-1 level in urine was positively correlated with disease activity assessed clinically by SLAM score. Higher level of urinary ICAM-1 was found among patients with severe activity p= 0.0018 when compared with patients with mild and moderate disease activity, but its level was higher in mild disease activity than in moderate disease activity. Conclusion: The significantly higher levels of adhesion molecules in patients with Lupus nephritis and its correlation with disease activity suggests that they can be used as a markers for lupus nephritis.  

30/30 PHENOTYPIC DETECTION OF METALLO- β-LACTAMASES PRODUCTION IN CLINICAL ISOLATES OF GRAM NEGATIVE BACILLI

Tahany Abdelhamid, Rasha A. Nasr, Mohamed S. Khater* and Amira Esmail

Medical Microbiology and Immunology, and *Geriatric Medicine Departments, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

In recent years, there has been an increase in the prevalence of carbapenem resistance among Gram negative bacilli which is acquired metallo-β-lactamases (MBLs), necessitating the rapid and accurate detection of these resistant strains in order to control their spread. This study aimed to compare imipenem-EDTA combined disc test (CDT), imipenem-EDTA double disc synergy test (DDST) and MBL E-test as convenient screening methods for detection of MBLs production in clinical isolates of Gram negative bacilli. A total of 115 nosocomial isolates of Gram negative bacilli from various clinical samples were subjected to susceptibility testing against various antibiotics using disk diffusion test. Minimal inhibitory concentration of imipenem by microbroth dilution method was also performed for all the isolates. Imipenem resistant isolates were further examined by imipenem-EDTA CDT, imipenem-EDTA DDST, and MBL E-test. Fifteen randomly selected imipenem susceptible isolates were as well tested for MBL production by CDT and DDST. Out of the 115 isolates of Gram negative bacilli, 31 (27%) showed resistance to imipenem; 16 isolates of P. aeruginosa, 12 A. baumannii, 2 E. coli and 1 Klebsiella spp.. The imipenem resistant isolates were isolated from wound infections (29%), followed by urine and lower respiratory tract samples equally (22.6%) then burn exudates (16.1%), blood samples (6.5%) and purulent pleural effusion sample (3.2%). All the 31 imipenem resistant isolates gave positive results with both MBL E-test and CDT (100% agreement), while DDST gave positive results in 14 (all were P. aeruginosa isolates) out of the 31 isolates (percentage agreement 45.2%). None of the 15 randomly selected imipenem susceptible isolates gave positive results by either DDST or CDT. In conclusion, CDT is superior to DDST in detection of MBLs and it is equally effective to MBL E-test for MBL detection in Gram negative bacilli, but given the cost constraints of the E-test, CDT can be used as a convenient screening method in microbiology laboratories.

31/30 CHARACTERIZATION OF PINK PIGMENTED FACULTATIVE METHYLOTROPHIC BACTERIA ISOLATED FROM PHYLLOSPHERE OF SOME PLANTS

Enas A. Hassan and Selim S. Ashoor

Agricultural Microbiology Department, Faculty of Agriculture, Ain Shams University, Shoubra El-Kheima, Cairo, Egypt.

One hundred eighteen pink-pigmented methylotrophic bacterial isolates were successfully isolated in pure form from the phyllosphere of eleven economical crops. The ability of these isolates to grow in presence of different methanol concentrations (0.5, 1.0, 1.5 & 2%) as a sole source of carbon was evaluated. They were also tested for their capability to accumulate and produce poly-β-hydroxy butyrate (PHB) and indole acetic acid (IAA). Results indicated that, among the tested isolates, only eighteen ones were able to grow well at all the tested methanol concentrations and have a good capability to accumulate and produce the PHB and IAA. Quantitative determinations of PHB, IAA and Cytokinin as well as urease activity of these cultures proved that the isolates namely CP3and M2gave the highest values. The morphological and physiological studies for the later isolates indicated that these strains are related to members of the genus Methylobacterium. A comparative 16S rRNA gene sequence-based phylogenetic analysis placed the strains in a clade with the species Methylobacterium extorquens and Methylobacterium zatmanii, with which it showed sequence similarities of 98% and 97 %, respectively.

32/30 MAGNITUDE OF GROUP B STREPTOCOCCUS CARRIAGE IN A SAMPLE OF EGYPTIAN PREGNANT WOMEN

Manal Darwish and Moustafa Ibrahim*

Microbiology and Immunology, * Obstetrics and Gynecology, Faculty of Medicine, Ain Shams University

To estimate the magnitude of Group B.streptoccus (GBS) colonization among a sample of pregnant women, and to evaluate the culture versus polymerase chain reaction (PCR). Diagnostic test accuracy study. Faculty of Medicine, Ain Shams University, Egypt.A total of 100 pregnant women at 35-37 weeks of gestation were attending Ain Shams University Maternity Hospital for antenatal care provided rectovaginal swab for testing. A total of 200 rectovaginal swabs were taken from 100 pregnant women. For each subject, the first swab was inoculated on chrom agar® strepto B (CA) with and without Lim broth enrichment, with subculture of the turbid bottles onto sheep blood agar. CA was examined for mauve colonies, Beta hemolytic (β. Hemolytic) and non hemolytic colonies were picked from sub cultured blood agar for further identification. The second swab was used for GBS detection by polymerase chain reaction using BD Gene Ohm® Strep B assay which is a qualitative in vitro diagnostic test for the rapid detection of GBS DNA in vaginal and rectal specimens. The primary outcome was GBS carriage rate among a sample of pregnant women in Egypt. The secondary outcomes were sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and overall accuracy of CA (with and without Lim broth enrichment) and PCR.The overall GBS colonization rate was 18%.Direct plating of the rectovaginal swab on CA resulted in detection of 17(17%) positive cases (carriers), whereas plating on a sheep blood agar after Lim broth enrichment (followed with full identification) yield 18(18%) positive cases. , PCR BD Gene Ohm tm strep B assay procedure was able to detect 19(19%) positive cases.The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the direct plating on CA media comparable to that of plating on a sheep blood agar after lim broth enrichment followed by full identification were 94%, 99%, 94% and 99%respectively. PCR BD Gene Ohm tm strep B assay procedure was able to detect 19% GBS cases. In addition that its sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in comparison to direct plating on CA media were 100%, 98%, 89% and 100% respectively. GBS carriage rate in the present study sample was 18% and combination of direct plating on CA with confirmation of CA positive isolates by means other tests is highly sensitive, specific and cost effective to detect GBS in pregnant women. Also, rapid PCR test should be targeted to women who presented in labor with an unknown GBS status.

33/30 PHENOTYPING AND GENOTYPING OF MRSA ISOLATES OBTAINED FROM HOSPITAL AND COMMUNITY ACQUIRED INFECTIONS IN A TERTIARY HOSPITAL IN QATAR DURING 2008-2010

Noha G. Khalaf, Mohamed El Ahmady* and J. Klena**

Department of Microbiology and Immunology, Faculty of Pharmacy, Modern Science and Arts (MSA) University, Egypt; ­*Department of Microbiology and Immunology, Faculty of Medicine, Zagazig University, Egypt; **CDC, Int. Emerg. Infect. Pgm., Beijing, China

Infections with S. aureus acquired in hospitals affect the patient health status, delay patients’ recovery and disturb the economic balance of healthcare institution. S. aureus infections acquired among community members usually cause serious skin infections and necrotizing pneumonia. We aim to identify S. aureus isolates that infect multinational patients admitted to different wards in a hospital in Qatar. A total of 750 S. aureus were obtained from inpatients and outpatients attending a tertiary care hospital in Doha, Qatar during 2008-2010. Bacterial identification was done manually; coagulase test was done using Staphaureux plus kits. MICs of different antibiotic classes were tested using VITEK 2 (bioMerieux, Inc. Durham, NC). Of the 750 isolates, MRSA isolates collected from skin infections were retained for this study. Methicillin resistance was confirmed with cefoxitin screening disc test and Latex agglutination test. Isolates resistant to erythromycin were tested for inducible clindamycin resistance using double disk test. Strain relatedness was investigated using macro- restriction analysis with PFGE. One hundred and ten MRSA isolates were identified, of which 86% were obtained from patients admitted to the emergency room. Thirty-two MRSA isolates were selected for further typing. Out of the 32 MRSA isolates, 40% were resistant to erythromycin. Cross resistance between erythromycin and clindamycin was seen. Resistance to mupirocin, tetracycline and sulfamethoxazole/trimethoprim was also observed. PFGE differentiated the 32 MRSA isolates into eleven pulsotypes, which was relevant to patients’ history and geographical origins. A rate of15% detection of MRSA was noted at the Qatar healthcare institute under study. These MRSA isolates were community and hospital acquired. A good correlation of PFGE data and clinical history of patients and to a lesser extent the antibiogram typing was observed. Common MRSA strains were spread among hospitalized patients and community members. Identical clones from clinically related patients points out to the mobility of these strains.

34/30 RAPID DETECTION OF METHICILLIN–RESISTANT STAPHYLOCOCCI FROM BLOOD CULTURE BOTTLES BY USING MULTIPLEX PCR ASSAY

Awany A.Gawish, Souzan H. Bekeer, Mai M. Helmy and Emad M. EL-Sayed

Department of Microbiology & Immunology, Faculty of Medicine,

Zagazig University

Methicillin resistant staphylococcal (MRS) infections represent a major nosocomial clinical problem that is associated with significant morbidity and mortality, especially in patients with bacteremia; so, rapid detection and accurate identification of MRS are critical for effective management of these infections. The purpose of this study: was to evaluate a method for rapid and direct detection of MRS, allowing clinicians to make prompt and effective decisions for the management of patients with staphylococcal bacteremia. The present work included: 100 patients with provisional diagnosis of bacteremia and twenty non bacteremia people as control group. For each person the following was done: full history taking, full clinical diagnosis, conventional blood culture for isolation of the causative organism and its identification by standard methods, multiplex polymerase chain reactions using 16S rRNA, nuc, mecA genes directly from positive blood culture bottles and from bacterial colonies. The result of this study showed that: 85 (85%) of those diagnosed as having or suspected to have bacteremia had given positive blood culture of which 85.8% were staphylococci and 41.2% were gram negative bacilli. Forty percent of staphylococci were found to be methicillin resistant by oxacillin disc diffusion and mecA by PCR. Oxacillin disc diffusion method had sensitivity, specificity, positive and negative predictive values of 95.2%, 87.5%, 97.5%, and 77.8%; respectively in relation to mecA detection by PCR. Significant agreement was found between direct multiplex PCR testing on blood culture bottles and PCR testing on bacterial isolates. IN conclusion: our results showed that rapid and direct detection of MRS is possible, allowing clinicians to make prompt and effective decisions for management of patients with staphylococcal bacteremia, and this is allowed by multiplex PCR.

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