Vol. 41, May, 2015

viagra quelle dose 1/41 PHENOTYPING AND RAPD-PCR-BASED GENOTYPING OF NOSOCOMIAL ACINETOBACTER BAUMANNII ISOLATES FROM INTENSIVE CARE UNITS IN EGYPTIAN HOSPITALS.

Moselhy S. Mansy, Mahmoud M. Tawfick, Osama H. Ahmed* and Fathy M. Elkady*

Microbiology and Immunology Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.

*Department of Intensive Care Unit, Faculty of Medicine, Al-Azhar University, Cairo, Egypt.

viagra pour les jeunes Acinetobacter baumannii have become one of the most important opportunistic pathogens causing nosocomial infections worldwide; especially among immunocompromised and critically ill patients. acheter viagra en ligne sans ordonnance A. baumannii are widespread in nature thus typing is needed to differentiate strains in the epidemiological studies. In this study, a total of 77 cialis 5 mg est il efficace A. baumannii isolates were recovered from 432 diverse clinical specimens with an isolation rate of 17.8 %. The specimens included in this study were collected from patients admitted to the intensive care units (ICUs) in four Egyptian hospitals, El-Sheikh Zayed, Dar-El Fouad, EL-Demerdash and Sayed Galal hospitals, during the period from October 2013 to December 2014. avantage viagra A. baumannii isolates were identified based on their cultural characteristics, biochemical reactions and API 20NE system, in addition to cialis et troubles de la vision blaOXA-51-like gene-directed PCR. Investigation of the antimicrobial susceptibility patterns of all isolates was performed using Kirby Bauer disc diffusion method. A. baumannii isolates in this study were highly resistant to diverse classes of antimicrobial agents including fluoroquinolones, aminoglycosides and cephalosporins. kamagra sachets A. baumannii isolates were phenotyped using biotyping based on API 20NE system and antibiogram typing, in addition to RAPD-PCR based genotyping. The results of the API 20NE system revealed six different biochemical profiles (biotypes) depending on the differences in the ability of A. baumannii isolates to utilize different carbon sources. In addition, 11 antibiogram types were detected among the isolates investigated. PCR-based RAPD analysis revealed 43 different RAPD-PCR based fingerprints among the isolates. In conclusion, this study revealed that almost the ICUs patients included in this study were infected with multi-drug resistant (MDR) mode emploi viagra 50mg A. baumannii. Biotyping alone or in combination with antibiogram has been shown to be a valuable method for initial screening to determine strain relatedness. These two methods were distinguished between anwendung cialis 20mg A. baumannii isolates and they did not always correlate. By comparing of RAPD-PCR to biotyping and antibiogram typing methods, our results indicated that RAPD-PCR technique represents a rapid, sensitive, reproducible and relatively simple mean with high discriminatory power for typing of le viagra generique est il efficace A. baumannii isolates over other two typing method.

 

 

2/41 FLUOROQUINOLONES RESISTANCE AND PLASMID PROFILE OF BACTERIAL ISOLATES RECOVERED FROM PULMONARY INFECTIONS

Abdel-Nasser A. El-Moghazy, Moselhy S. Mansy, Sahar M. Ramzy* and

Kareem A. Ibrahim**

Microbiology & Immunology Department, Faculty of Pharmacy (Male), Al-Azhar University, Cairo, Egypt

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

**Microbiology & Immunology Department, Faculty of Pharmacy, Egyptian Russian University, Cairo, Egypt

Widespread uses of fluoroquinolones (FQs) have resulted in increasing incidences of resistance to these agents. The aim of this study is to assess the susceptibility of bacterial isolates recovered from patients with pulmonary infection to FQs and to analyze the plasmid profile of the highly resistant isolates. A total of 508 sputum samples were collected and identification of 497 isolates was done using conventional method and Microscan. Antimicrobial susceptibility testing of 455 bacterial isolates was performed by disk agar diffusion method and Microscan. The major bacterial isolates were Klebsiella pneumoniae (25.3%), Streptococcus pneumoniae (14.9%) and Escherichia coli (14.7%). The highest resistance observed was to Nalidixic acid (78%) and Norfloxacin (16%). The lowest resistance observed was to the third generation FQs (11% to each). Some Klebsiella pneumoniae and Escherichia coli isolates showed resistance to all eight FQs agents employed in this study and were subjected to plasmid profile analysis, which showed different bands in all of them ranging from 1.5 to >20 kb and more than half of them showed common bands at 3, 4 and 5 kb.

 

3/41 STUDIES ON RESISTANCE MECHANISMS IN MEROPENEM RESISTANT ACINETOBACTER SPP.

Sameh M.M. AbdelGhani, Walid Bakeer, Sayed Ali Galal and Magdy Ali Amin*

Microbiology & Immunology, Faculty of Pharmacy, Beni-Suef University, Egypt.

*Microbiology & Immunology, Faculty of Pharmacy, Cairo University, Egypt.

Meropenem is considered one of the mostly used carbapenems efficiently against resistant infections in Egypt. Acinetobacter constitutes one of the most serious infectious agents that developed resistance against a diversity of efficient antibiotics like carbapenems. Out of 637 clinical isolates obtained from different health care provenance, 104 pure isolates were meropenem resistant which were identified as following; 54 Acinetobacter spp., 32 Pseudomonas aeruginosa, 3 Pseudomonas fluorescence, 12 Klebesiella spp., 12 Enterobacter spp., 1 Vibrio fluvialis. The meropenem resistant Acinetobacter spp., were chosen for further studies. The modified Hodge test resulted in 39 (72%) isolates exhibited carbapenemase activity. EDTA disc-synergy test showed 13 (24%) isolates have metallo-β-lactamases. AmpC induction test showed only one (2%) isolate was positive. Two (4%) isolates only showed resistance to meropenem due to efflux mechanism this evidenced by screening using omeprazole and verapamil. Twenty isolates were harboring the blaoxa-51-like gene while 13 (65%) isolates of them were carrying the blaoxa-23-like gene. In this study, OXA carbapenamases in meropenem resistant Acinetobacter spp. were detected, blaOXA-51 (n = 20) and blaOXA-23 (n = 13), they coexisted in 13 isolates, while blaOXA-51 was found alone in 7 isolates. The coexistence of blaOXA-23 like, blaOXA-24 like, blaOXA-51 like and blaOXA-58 like genes has been reported, especially that of blaOXA-23 and blaOXA-51. From this study we found that many mechanisms were involved in meropenem resistance. Carbapenemases production were the main reason for the studied Acinetobacter spp. resistance towards meropenem.

 


4/41 DETECTION OF GYRA AND PARC MUTATIONS IN FLUOROQUINOLONES HIGHLY RESISTANT KLEBSIELLA PNEUMONIAE AND ESCHERICHIA COLI ISOLATES RECOVERED FROM PULMONARY INFECTIONS

Moselhy S. Mansy, Abd El-Nasser A. El-Moghazy, Sahar M. Ramzy* and Kareem A. Ibrahim**

Microbiology & Immunology Department, Faculty of Pharmacy (Male), Al-Azhar University, Cairo, Egypt

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

**Microbiology & Immunology Department, Faculty of Pharmacy, Egyptian Russian University, Cairo, Egypt

Widespread uses of fluoroquinolones (FQs) have resulted in increasing incidences of resistance to these agents which rings the alarming for FQs are becoming useless within few years. The aim of this study was to detect the presence of gyrA and parC mutations in Klebsiella pneumoniae and Escherichia coli isolates recovered from pulmonary infection in Abbassya Chest Hospital and National Cancer Institute. A total of 49 highly resistant isolates were recovered from more than 500 sputum samples and were identified using conventional method and Microscan technique. Antimicrobial susceptibility profiles were investigated using Microscan technique. These highly resistant isolates were resistant to at least one quinolone agent from each generation. They were tested against 9 agents: nalidixic acid, ofloxacin, norfloxacin, pefloxacin, ciprofloxacin, levofloxacin, moxifloxacin, gatifloxacin and gemifloxacin. Detection of gyrA and parC mutations was done using duplex polymerase chain reaction (dPCR) that showed parC mutation in 18 isolates (36.7%), gyrA mutations in only 6 isolates (12.2%), while isolates with both mutations were 9 isolates (18.4%) and isolates showed neither mutation were 16 isolates (32.7%). Mechanism of resistance in K. pneumoniae and E. coli isolates was mostly chromosomal mediated due to mutation in gyrA and parC subunits of DNA gyrase and topoisomerase 4 respectively. Presence of isolates with neither mutation suggests the resistance could be plasmid mediated and requires further investigation of the plasmid genes that could contribute to the high resistance to FQs.

 

5/41 ANTIMICROBIAL ACTIVITY OF SOME PROBIOTICS GROWN ON XYLOOLIGOSACCHARIDES

Mayada H. Aly, Salwa A. Megahed*, Mohamed A. Ramadan* and Mohamed M. El-Din Hussein**

Department of Crime Investigation Research, National Center for Social and Criminological Research, Cairo, Egypt

*Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Egypt

**Department of Natural and Microbial Products, National Research Center, Dokki, Cairo, Egypt

Xylooligosaccharides (XOS's) were produced from xylose-containing polysaccharides (XPS's) obtained from natural, xylan-rich, agro-industrial wastes, i.e., corncobs and sugarcane bagasse. XOS's were prepared by partial hydrolysis of XPS's either chemically by oxalic acid 0.25N or by fungal attack by Aspergillus niger and by Trichoderma reesei. This was followed by evaluation of the antimicrobial activities of some probiotics grown on the chosen XOS-mixtures. The studied probiotics included; Bifidobacterium bifidum, Lactobacillus acidophilus and Lactobacillus casei. Most of the culture filtrate of probiotics showed variable antimicrobial activities against isolated strains of Escherichia coli, Salmonella typhi and Staphylococcus aureus.

 

 

6/41 EARLY AND ACCURATE DIFFERENTIAL DIAGNOSTIC POTENTIAL OF TUBERCULOSIS USING ANTIGEN 85B AND ESAT-6 PROTEIN ANTIGENS

Yousef A. Soliman, Ramadan Al-Domany*, Nevine A. Abdelaziz***, Riham M. Shawky* and Shimaa. A. Hamied**

Central Laboratory for Evaluation of Veterinary Biologics, Abassia, Cairo, Egypt

*Department of Microbiology, Faculty of Pharmacy, Helwan University, Cairo, Egypt

**Vaccine and Sera Research Institute, Cairo, Egypt

***Department of Microbiology, Faculty of Pharmacy, University of Technology and Information, Cairo, Egypt

In the aim of developing new diagnostic reagents for tuberculosis, the culture filtrate, recombinant ESAT-6 and antigen 85B were prepared and their differential diagnostic potential was studied in the artificially infected guinea pigs with different mycobacterial strains. The results of both ELISA and cellular proliferation assays revealed that the culture filtrate can detect the mycobacterial infection but cannot differentiate between the M. tuberculosis infected animals from those sensitized with the atypical strains or vaccinated with the BCG vaccine. In contrast, the recombinant ESAT-6 Protein Antigen was able to detect only the truly infected animals with either M. tuberculosis or M. bovis where those infected with atypical strains or the vaccinated animals were negative. The current study proves the capability of using such antigen cocktail for accurate and differential diagnosis of tuberculosis.

 

 

7/41 EVALUATION OF INFECTION CONTROL MEASURES IN ACADEMIC DENTAL CLINICS

Mai A. Abdallah, Reham Wasfi, Sahar M. Radwan*, Mohamed S. Ashour**

Microbiology & Immunology Department, Faculty of Pharmacy, October University for Modern Sciences and Arts, Giza, Egypt.

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

**Microbiology & Immunology Department, Faculty of Pharmacy (Boys), Al-Azhar University, Cairo, Egypt.

Infection control is a primary mean of disease prevention in all healthcare settings, including the dental office. Dentistry is a profession that involves constant risk of exposure to various environmental and human infectious agents; contamination can affect both staff and patients. The infection control measures applied in the academic dental clinics in October University for Modern Sciences and Arts (MSA), Giza, Egypt, were evaluated. A total of 1300 samples were collected from personnel including dentists, nurses and patients, as well as environmental samples including air, surfaces and instruments. Bacterial isolates were identified and then tested for their antimicrobial susceptibility. Seven commonly used disinfectants in the dental clinics were tested for their efficiencies to kill the tested microorganisms via kill time suspension test in presence and absence of organic load, carrier test, sporicidal test and practical test. The tests included ethyl alcohol 70%, sodium hypochlorite 5% (Chlorox®), chloroxylenol (Dettol®), chlorhexidine 0.3%+cetrmide 3% (Setavlon®), povidone-iodine (Betadine®), hydrogen peroxide 3% and glutaraldehyde 2%. A total of 863 isolates were recovered from different samples. A total of 637 out of 1030 environmental samples (61.8%) showed positive growth of microorganisms while 83.7% (226/270) of personnel samples were positive for microbial growth. Gram positive bacteria were more predominant than Gram negative isolates. The most predominant Gram positive bacteria isolated in this study were coagulase negative Staphyloccus sp. (CoNS), followed by Bacillus species and the main Gram negative bacteria isolated in this study were identified as Klebsiella sp. followed by Pseudomonas aeruginosa and Enterobacter sp. Candida sp. represented 3.7% of total isolates. Gram negative and Gram positive isolates showed high resistance to erythromycin. Among recovered Staphylococcus aureus isolates about 46% were methicillin resistant. Twenty four bacterial and fungal isolates were included in the evaluation of disinfectant efficiency. The obtained results indicated that most of the tested disinfectant products were effective at the manufacturer recommended level within 30 min of contact time when tested in the absence of organic matter. However, when organic matter was present longer contact times were needed to demonstrate the effectiveness. Glutaraldehyde 2% had satisfactory antimicrobial activity even in the presence of organic matter, and high killing effect at contact time from 1 to 5 min with all tested isolates in addition to its effectiveness against bacterial spores.

 

8/41 INDUCTION OF MELANIN BIOSYNTHESIS IN KLUYVEROMYCES MARXIANUS

Sanaa Mohamed Ashour

Botany Department, Faculty of Women for Arts, Science and Education,

Ain Shams University, Cairo, Egypt

The objective of this study was to achieve more high production of melanin pigment by Kluyveromyces marxianus in the presence of L‑tyrosine and salt stress by standard methods, the production of melanin reached its maximum 14.3 g/l of the crude melanin when the cells grown in Yeast extract peptone xylose (YEPXyl) broth in the presence of 25 mM L‑tyrosine adding to 4% NaCl and 5 mM glycinebetain. The gene responsible for the production of melanin in K. marxianus may be chromosomally encoded.These resultsmake K. marxianus a promising candidate for a great biotechnological future in this field specially that melanin have immense application potentials in the field of agriculture, cosmetics and pharmaceutical industries.

 

9/41 ROLE OF RHIZOBIUM AND PHOSPHATE SOLUBILIZING BACTERIA TO ENHANCE THE GROWTH OF WHEAT AND REDUCE THE PHOSPHATIC FERTILIZERS

Amal A. Ali

Soils, Water and Environment Res. Inst., Agriculture Research Center, Giza, Egypt.

Rhizobium and phosphate solubilizing bacteria (PSB) are important to plant nutrition. These microbes also play a significant role as plant growth-promoting rhizobacteria (PGPR) in the biofertilization of crops and convert insoluble forms of phosphorus to an accessible form to increase phosphorus uptake and plant yields. In this study, the efficacy of Six effective bacterial strains of Rhizobium leguminosarum bv. Viceae strain 441, Rhizobium leguminosarum bv. trifolii (100), Bacillus megaterium, Bacillus polymyxa, Bacillus circulans and Pseudomonas fluorescens were tested for phosphate solubilization, IAA and ammonia production. Upon these tests Pseudomonas fluorescens, Bacillus megaterium and Rhizobium sp. Strain 100 were selected for further tests. Rhizobium combined with pseudomonas fluorescens or Bacillus megaterium were used in single or in combinationduring planting of wheat in saline soil. The results of field experiment revealed that dual inoculation of rhizobium with phosphate solubilizing bacteria significantly improved soil pH and total soluble salts (EC) which reflected on the crop growth and yield components where there was increasing in shoot weight, plant height and total chlorophyll in addition to grain yield, N, P contents, protein and carbohydrates.proline content was also studied.

 

10/41 THE IMPACT OF YEAST INOCULATION UNDER DIFFERENT RATES OF COMPOST AND BIOGAS MANURE ON WHEAT PLANTS GROWN UNDER SALT STRESS.

Hanaa A. Abo-Kora, Hager I. Tolba and El-Sayeda H.M. EL-Badawy.

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

A pot experiment was carried out during winter season 2014 to study the effect of Sacccharomyces cerevisiae combined with compost or biogas manure on the chlorophyll content, growth, antioxidant status and mineral uptake of wheat under soil salinity. Two sources of fertilizers for soil amendment: compost and biogas manure were used. Results indicated that, inoculation with Sacccharomyces combined with compost or biogas manure in saline soil alleviated the salt stress on the growth parameters, yield, total biomass and enzymatic activity. The inoculation with Sacccharomyces cerevisiae also induced a significant stimulatory effect on the chlorophyll pigments and carotinoids. Salinity tolerance measured as percentage chlorophyll stability index (CSI %) was higher under treatment inoculated with Sacccharomyces. Data also showed that treatment with Sacccharomyces combined with compost or biogas manure succeeded to increase contents of N, P, K of shoot of wheat plant compared to un-inoculated plants. Moreover, inoculation significantly reduced accumulation of Proline (amino acid) content in the shoots and roots, antioxidant enzymes (peroxidase and catalase) of the wheat plants under soil salinity. As a result, treatment with Sacccharomyces cerevisiae combined with compost or biogas manure could be alleviating the effect of potentially toxic ions. The un-inoculated plants were more sensitive to salinity than inoculated ones, as indicated by several measured parameters.

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