. Vol. 37, January, 2014

viagra effet positif 1/37 COMPARISON OF SOME VIRULENCE FACTORS AND ANTIMICROBIAL RESISTANCE ASSOCIATED GENES OF BIOFILM AND NON-BIOFILM PRODUCING PSEUDOMONAS AERUGINOSA

Ramadan Hassan, Rasha Barwa and Heba Adel*

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

*Talkha Central Hospital, Mansoura, Egypt.

viagra femme pas cher Pseudomonas aeruginosa produces multiple virulence factors that have been engaged in pathogenesis. The aim of this study was to compare between some virulence factors and antimicrobial resistance associated genes of biofilm and non-biofilm producing acheter cialis original 10mg Pseudomonas aeruginosaisolatedfrom Mansoura Hospitals. In this study, a total of 50 strains of viagra qui ne marche pas Pseudomonas aeruginosa were isolated from 150 clinical specimens collected from Mansoura Hospitals. The phenotypic detection of biofilm revealed that thirty two isolates (64%) were considered as positive biofilm producers. The largest numbers of positive samples were from urine samples. The production of exopolysaccharides, such as cialis duree efficacite PelA was required for prix du cialis 5mg en pharmacie france Ps. aeruginosa bioļ¬lm formation. Detection of achat generique viagra france pelA gene revealed that it was harbored by chromosomal DNA of all biofilm producing isolates. Associations were assessed between biofilm production and virulence associated genes including Exotoxin A, cialis original le moins cher lasB elastase and type III secretion system ( l'effet du kamagra exoSand comparatif cialis generique exoY). Investigation of virulence factors associated genes revealed that 81.25%, 69.23%, 69.7 % and 91.6% of the isolates harboring toxA, lasB, exoSand exoY genes respectively were biofilm producers. Regarding resistance associated genes, 67.44% of AmpC producing isolates were biofilm producers. While investigation of the tripartite efflux system MexAB–OprM reveled that it was present only in eight Ps. aeruginosa isolates and all of them were biofilm producers. The present study confirmed that antimicrobial resistance and virulence associated genes were more prominent in biofilm-producing Ps. aeruginosa than in non-biofilm-producers.

2/37 EVALUATION OF ANTIBACTERIAL ACTIVITIES OF SOME LACTIC ACID BACTERIA AGAINST DIFFERENT ENVIRONMENTAL SALMONELLA ISOLATES

Ahmed M. Ismail, Wael M. Tawakkol* andAbdel-Gawad M. Hashem*

Department of Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology, Cairo, Egypt

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

In the present study six lactic acid bacteria were recovered from dairy products were collected from different factories and identified by Analytical Profile Index (API) 50CH system. The antibacterial activities of both cell free supernatants and protein precipitates from these six isolates against 21 environmental Salmonella isolates were evaluated. Extraction of protein from lactic acid cell-free-extract was done using ammonium sulphate precipitation method. Lactobacillus paracasei and Lactobacillus rhamnosus had the strongest activities against the tested isolated Salmonella. It is concluded that the antimicrobial inhibitory effect was the most probably related to the supernatant of cell free extract.


3/37 DIFFERENT DIAGNOSTIC METHODS IN CASES OF BRUCELLOSIS

Ayman Asaad, Lamiaa A. Adel and Enas H. Allam*

Medical Microbiology & Immunology Department, Tropical Medicine Department*, Faulty of Medicine, Ain Shams University

Brucellosis is a worldwide zoonosis transmitted to human from infected animals with a high degree of morbidity in human. As the clinical finding is non-specific, diagnosis is difficult and may be easily missed; specific diagnosis requires isolation of the causative organism, demonstration of high level of specific antibodies or seroconversion. Evaluation of different diagnostic tests; blood culture, ELISA, slide and tube agglutination and polymerase chain reaction in diagnosis of brucellosis and find the agreement between these diagnostic methods. Forty patients were included in this study; they were presented with fever of unknown origin but clinical data suggestive of brucellosis. They were divided into two groups; twenty patients with negative slide agglutination test for brucellosis (group I) and twenty patients with positive slide agglutination test (group II). The blood culture was positive in 30% of patients with positive slide and tube agglutination tests and none of patients with negative slide and tube agglutination test. The results of slide agglutination test were the same as the tube agglutination test. There was no statistically significant difference between the result of ELISA and slide agglutination test or tube agglutination tests. The ELISA and PCR tests were positive in 90% and 60% of patients with positive agglutination tests respectively and were negative in all of patients with negative agglutination tests. There was highly statistically significant difference between the result of ELISA and both blood culture and PCR results.The slide agglutination test remains an easy, fast, reproducible test for the laboratory diagnosis of brucellosis. The role of blood culture is still limited and time consuming.

4/37 ROTAVIRUS PREVALENCE AMONG DIARRHEIC CHILDREN UNDER 5 YEARS OLD IN EL-BEIDA CITY, EL-JABAL AL-AKHDAR, EASTERN NORTH OF LIBYA

Salha Farag Ben-Gweirif, Salwa Ibrahiem El –Tawaty* and Maha Eid Omran**

Botany Department, Faculty of Science, Benghazi University, Benghazi, Libya.

*Department of Microbiology & Immunology, Faculty of Pharmacy, Omar Al-Mukhtar, University, El-Bieda, Libya.

**Department of Microbiology & Immunology, Faculty of Pharmacy (Girls),

Al-Azhar University, Egypt.

Rotavirus gastroenteritis (RVGE) is the most common cause of severe childhood diarrhoea worldwide. Previous studies in different areas of Libya showed that rotavirus is the leading cause of infectious diarrhoea. This is the first systematic study for assessment of the prevalence of rotavirus in diarrheic children under 5 years old, in El-Beida city, El-Jabal al-Akhdar, the eastern north district of Libya. A total of 200 stool samples from children who were attending the Paediatric Department in El-Thoura Hospital in El-Beida city were tested for the presence of Rotavirus Antigen by ELISA and examined for enteric pathogenic bacteria by the conventional methods. Antimicrobial susceptibility was done by disk diffusion method and Microscan automated system. Rotavirus was the most prevalent agent 58.3%, followed by diarrhoeagenic E. coli 25% and Salmonella spp.16.7%. The rotavirus detection rate was higher in males 67.9% than in females 32.1% meanwhile 82.1% of rotavirus positive cases were aged 12 months or less. Among the examined children only 7.1% were on breast feed, while 57.1% on bottle feed and 35.8% on mixed feed. In this study all E. coli and Salmonella spp. isolated were sensitive (100%) to Augmentin, Amikacin and Cephalexin, while they showed high resistance rate to Ampicillin and Trimethoprim-Sulphamethoxazole. Both E. coli and Salmonella isolates showed comparable resistance to Tetracycline. As conclusion, detection of rotavirus antigen should be taken into consideration in routine diarrhoea investigations in El-Beida city, besides application of vaccination programme against rotavirus for infants may lessen the RVGE burden in El-Beida city and an education programme for mothers about the importance of breastfeeding as a parameter for protection against enteropathogens.

5/37 Review Article

IMPACT OF ACQUIRED RESISTANCE ON UROPATHOGENICESCHERICHIA COLI. PATHOGENICITY

Miran Yousri El Sayed El Far, Ossama Mohammad El Tayeb*and Mohammad Mabrouk Aboulwafa**

Department of Microbiology and Immunology, Faculty of Pharmacy, Ahram Canadian University, 6th October, Egypt

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

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

Antimicrobial resistance is one of the most important challenges,whichmay face health professionals and the public. The rate and the extent of antimicrobial resistance spread vary widely with different pathogen-drug combinations.These depend on the appearance of resistant variants and the intensity of selection imposed by the antimicrobial treatment. The ability of a microbial pathogen to cause infection is dependent on its fitness and virulence towards the host. The microorganism capacity for gaining novel resistance genes plays an important role in bacterial adaptive evolution. Unfortunately, compensatory evolution for the lost fitness and reduced virulence if incurred, may lead to clonal spread of the fittest resistant microbial pathogens.Widespread resistance in enteric bacteria is a particular problem in less-developed areas of the world. Heavy and indiscriminate use of antibiotics may combine with a high prevalence of drug-resistant bacteria in the fecal flora, poor standards of sanitation, and a high incidence of diarrheal disease. These will in turn, encourage the rapid emergence and spread of multi-resistant strains of enteric bacteria. In a recent study conducted in Egypt, it was found that a biological cost due to antimicrobial resistance may be exerted and affect virulence determinants and bacterial fitness. There was no relationship between the level of the biological cost exerted by mutants and their MIC values. Mutants showed lowered adherence, lower abilities of in vitro biofilm formation and lowered abilities to secrete cell free hemolysins and cytotoxicity.

6/37 PHENOTYPIC AND MOLECULAR DETECTION OF EXTENDED-SPECTRUM Β-LACTAMASE PRODUCING ACINETOBACTER BAUMANNII AMONG IMMUNOCOMPROMISED PATIENTS.

Fatma Al-Zahraa M. Gomaa, Wael M. Tawakol and Fatma I. Abo El-Azm

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

Department of Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology, Cairo, Egypt.

A. baumannii has emerged in the healthcare settings as one of the most troublesome pathogens both globally and locally. Extended-spectrum β-lactamases (ESBLs) is one of the important groups of β-lactamases.The aim of this work is to investigate the prevalence of Extended Spectrum β-lactamases in clinical Acinetobacter baumannii isolates and to evaluate the sensitivity of manual, semi automated Microscan System and Polymerase chain reaction in detecting ESBLs production. Also, to determine the most common source of this resistant bacteria. A total of 60 Acinetobacter baumannii isolates were included in the study. Isolates were identified conventionally and using Microscan semi-automated identification system. Antimicrobial susceptibility was determined for all isolates by agar plate disc diffusionmethod and using Microscan semi-automated identification system. Potential ESBLs production was investigated among 45 multidrug-resistant (MDR) Acinetobacter baumanniiisolates using double disk synergy test (DDST), Microscan and conventional PCR. Acinetobacter baumannii isolates were more frequently implicated in blood stream and respiratory tract infections. MDR A. baumannii represent 75% of isolates. All tested MDRAB were producing ESBLs as determined by Microscan system and PCR whereas only 20% of ESBL producing isolates were determined by using double disk synergy test (DDST). Multidrug resistant A. baumannii became a problematic organism in immunosuppressed patients. The identification of ESBLs by conventional phenotypic methods remains sometimes difficult in practice. PCR could be considered as a sensitive and specific reference method for detection of ESBL producing A. baumannii. The Microscan proved to be a valuable tool with high sensitivity for ESBL detection. Blood and sputum could be a source for emergence and spread of ESBL producing A. baumannii among immunocompromised patients.

7/37 CHEMOKINE RECEPTOR4 AS A POSSIBLE MARKER OF SEVERITY IN BRONCHIAL ASTHMA

Nehad M. Sayed, Alaa A. Aly and Nehad M. Osman*

Microbiology and Immunology Department and Pulmonary Medicine Department*

Faculty of Medicine, Ain Shams University

Asthma is a serious global health problem affecting all age groups. The CC-chemokine receptor 4(CCR4), and its ligands may play key roles in T cell chemotaxis into the bronchial tissues in asthma. So blocking CCR4-ligand interaction may provide a novel therapeutic approach in allergic disease. The present work aimed to study the clinical relevance of CCR4+CD4+T cells as a possible diagnostic and prognostic marker of severity in bronchial asthma to open new therapeutic approaches. This casecontrol study enrolled a total number of 80 non–smoking subjects divided in two groups. Sixty asthmatic patients of variable severity and twenty age and sex matched healthy control subjects. Diagnosis of asthma and assessment of the severity and treatment were based on clinical symptoms and the criteria of Global Initiative for Asthma (GINA 2012) guidelines.The patients were categorized according to severity into 3 groups, mild asthma (15 patients), moderate asthma (15patients) and severe asthma (30 patients). The response to treatment was assed according to clinical and functional parameters. All individuals were subjected to the followings: full history, thoroughly clinical examination, pulmonary functions tests, and analysis of CCR4 expression on peripheral blood CD4 T cells by Flow cytometry for each subject during acute asthma attack on day 1and after receiving treatment on day 7.The present study showed that all the studied patients were responding to treatment except three cases from patients with severe asthma. Comparison between the studied groups as regards CCR4+CD4+T cells showed that patients with severe bronchial asthma had the highest baseline values of blood CCR4+ T cells (53% ± 1.5), followed by those with moderate asthma (46% ± 1.8%),and mild asthma ( 40.2% ± 1), while control subjects had the least values (28.5% ± 2.5%), and the difference was statistically highly significant. The percentages of CCR4+ T cells decreased significantly after treatment in the responders of the three asthmatic groups. The non-responders showed higher values of blood CCR4+ T cells before and after treatment (54.3± 1.9 and 58.6 ±1.5)   compared to responders (47.7±5.6 was41.9±4.3). In this study the CCR4 expression was perfect and reliable in predicting asthma severity. The best cut off value was 36.6% for mild, 38.4% for moderate and 42.4%for severe cases with 100%, sensitivity, specificity 100%, and diagnostic accuracy of 100%. High CCR4 expression on T cells is an important biomarker of severity in bronchial asthma and could be readily used as an indicator of bad response to treatment. Further clinical trial are required to validate the concept that targeting CCR4+T cells would have a significant effect on airway inflammation in asthma, and also to help selecting the patient subpopulation in which a CCR4 antagonist is likely to work best.

 

 

8/37 SACCHAROMYCES CEREVISIAE LIPASE: CHARACTERIZATION AND NEMATICIDAL ACTIVITY AGAINST MELOIDOGYNE JAVANICA

Mohsen A. Sayed

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

Saccharomyces cerevisiae was isolated from oil waste samples on a selective medium containing tributyrine as a carbon source. The isolates were cultivated for lipase production in shake flasks containing 1% Tween 80 as sole carbon source at 37oC. The enzyme was partially purified by (NH4)2SO4 precipitation and dialysis. Seven yeast isolates were screened from oil waste samples. Four of the isolated yeasts exhibited lipolytic activity. The most potent isolate was identified as Saccharomyces cerevisiae and was selected for optimization of lipolytic activity. Maximum activity (9.1 U ml-1) was obtained at temperature 40°C and pH 7. Ca2+ (1 mM) enhanced lipase activity followed by Mg2+ and Zn2+, at the same concentration. The enzyme was completely inhibited by both Hg2+ and Cd2+. S. cerevisiae lipase showed low specificity as it could hydrolyze olive oil, tributyrine, tween 80, tween 40 and tween 20. Nematicidal activity assay revealed that the enzyme caused 53% inhibition in mobility of adult Meloidogyne javanica and decreased egg hatching by 74%.

9/37 APPLICATION OF PLACKETT–BURMAN SCREENING DESIGN TO MODEL THE CULTURE MEDIUM USED FOR PENICILLIN G ACYLASE PRODUCTION BY ARTHROBACTER VISCOSUS NRRL B-1973.

Magdy M. M. Elnashar1,4,5, Marwa I. Wahba2,4, Magdy A. Amin3, and Ahmed I. Eldiwany2

1Polymers Department, National Research Center, El-Behooth St., Dokki, Cairo, Egypt.

2 Department of Natural and Microbial Chemistry, National Research Center, El-Behooth St., Dokki, Cairo, Egypt.

3Microbiology and immunology Department-Faculty of Pharmacy–Cairo University.

4Centre of Scientific Excellence-Group of Biopolymers and Nanobiotechnology, Cairo, Egypt.

5College of Medicine, Taif Univ., Hawiya, Taif, KSA

In the present study, the Plackett-Burman design (PBD) was used to screen the effect of 19 factors on the production of penicillin G acylase (PGA) from Arthrobacter viscosus NRRL B-1973. The results of the PBD showed a 4.9 fold variation in the amount PGA produced from 5.89 to 28.9 U/ml, reflecting the importance of the optimizing process. Among the nineteen tested factors, only six were proven to be significant. These were; tryptone, arginine, maltose, and dextrin, which exerted a significant positive effect on the PGA production, and MgSO4 and NaCl, that offered significant negative effects.

10/37 CORRELATION BETWEEN MICRODILUTION METHOD AND THREE COMMERCIALLY AVAILABLEMETHODS FOR ANTIFUNGAL SUSCEPTIBILITY TESTING OF CANDIDA SPECIES ISOLATED FROM IMMUNOCOMPROMISED EGYPTIAN PATIENTS

Shereen Atef Ammar, Wafaa Nabil El-Tayeb and Abd El-Gawad M. Hashem*

Microbiology Department, Faculty of Pharmacy, Misr International University

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

The correlation between broth micro dilution, E-test, disc diffusion, and fungifastAFG was determined for fluconazole, and amphotericin B. The minimal inhibitory concentration of those antifungal agents was established for fifty Candida spp. isolates collected from immunocompromised patients admitted to the National Cancer Institute, Cairo, Egypt. The identified species were: C. albicans, C. glabrata, C. tropicalis and C. krusei. Non-Candida albicans showed higher minimal inhibitory concentration (MIC) when compared with C. albicans especially with fluconazole. The overall concordance (based on the MIC value obtained within two dilutions) between broth microdilution and E-test was 83.3% and 70% for fluconazole, and amphotericin B respectively. While, the overall concordance between broth microdilution and fungifastAFG was 62.5% and 100%for fluconazole, and amphotericin B respectively. Considering the categorical agreement between the disc diffusion and broth microdilution method showed the highest rates of agreement for amphotericin B (96%), followed by fluconazole (60.78%), the decreased level of agreement seen with fluconazole was largely due to false resistant results. Regarding categorical agreement between broth microdilutionand fungifastAFG was 70.8% and 100% forfluconazole and amphotericin B respectively. Commercially available testing techniques correlates well with broth microdilution, yet further research are required to standardize these methods to include all antifungal agents.

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