Vol. 29, May, 2011

acheter viagra medicament 1/29 ASSESSMENT OF HUMORAL IMMUNITY IN CHILDREN WITH AUTISM

Abeer A. El-Sayed and Marwa A. El-Missiry*

Departmentsof Medical Microbiology & Immunology and   Neuropsychiatry*, Faculty of Medicine, Ain Shams University, Cairo, Egypt

Autism is a behaviorally defined disorder, which is the endpoint of several organic etiologies .The number of children diagnosed as having   autism is increasing for various reasons. There is growing evidence that an abnormal immune response may exert a negative influence on neurodevelopment, potentially contributing to the etiology of some cases of autism. quel site pour acheter du viagra The current study was designed toassess humoral immunity in autistic patients by evaluating IgM and IgG serum levels compared to matched controls and to correlate antibody levels with autistic syptomatology. ou acheter du viagra a quebec Subjects & Methods: The study was enrolled on 48 children divided into 2 groups. Consent was taken from parents before starting. Cases (n=24): autistic children attended Institute of Childhood Postgraduate Studies, Ain Shams University, from October 2010 till March 2011.Controls (n=24 ): age and sex matched typically developed children. All children were subjected to a full history taking and rating scales e.g.{Gilliam Autism Rating Scale or Vineland Adaptive Behavior Scale}.Three ml of blood were collected by venepuncture under complete aseptic conditions. Serum level IgM and IgG were measured using quantitative Nephelometry. cialis de marque The result of the present study revealed that autism was more frequent in boys than girls as 24 autistic children were 20 males (83.3 %) and 4 females (16.7%). Children with autism had higher levels of IgM & IgG compared to controls with highly statistically significant difference (p viagra personne normal < 0.01).However, no statistically significant correlation was detected between either IgM or IgG (on one side) and   autistic symtomatology (on the other side). cialis et prostate Conclusion: Immune system was dysreguated in autism, as evidenced by elevation in serum levels of IgM and IgG in autistic children than normal controls. So, immune system may have a potential role in etiology of autism.      

dapoxetine effet 2/29 A MOLECULAR STUDY OF SULPHONAMIDES RESISTANCE AMONG ESCHERICHIA COLI ISOLATED FROM URINE

Abeer Ghazal, Eglal El-Sherbiny, Hamida Abou-Shleib*, Nourhan Fanaki* and Marwa Atef**

Microbiology Department, Medical Research Institute, Alexandria University  

*Pharmaceutical Microbiology Department, Faulty of Pharmacy,

Alexandria University

**Microbiology and Immunology Department, Faulty of Pharmacy,

Pharos University

The aim of this study was to describe the distribution of the sulfamethoxazole resistance genes and their association with class 1 integrons in uropathogenic risque viagra périmé Escherischia coli isolates. This study included 50 sulphonamide resistant acheter priligy sans ordonnance E. j'ai pris du kamagra coli isolates that were collected from 74 clinical specimens.Resistance to sulfamethoxazole was assessed by the disk diffusion method and broth dilution method. Polymerase chain reaction (PCR) with primers specific for sul1, sul2, sul3 and int1 was used to detect the three known sulphonamide resistance genes. The sul1 gene was found in 15 (30%) isolates, sul 2 gene in 16 (32%) isolates, and both genes in 8 (16 %) isolates. The sul3 gene was found in 1 (2%) isolate. The int1 gene was found in 2 out of 15 (13.3%) of the sul1 positive isolates.

3/29 IN VITRO ACTIVITY OF ERTAPENEM AGAINST BACTERIAL ISOLATES FROM DIABETIC FOOT INFECTION IN AIN SHAMS UNIVERSITY HOSPITAL

Sara R. Abd El-Halim, Lamiaa A. Adel, Abeer A. Elsayed and Mohamed E. Elserafy*

Medical Microbiology and Immunology,   *General Surgery Departments, Faculty of Medicine, Ain Shams University

Background; Foot infection occur relatively frequent in individuals with diabetes, almost always following trauma, and dramatically increase the risk of hospitalization and amputation. The diabetic individuals have many risk factor known to be associated with lower extremities complications (e.g. peripheral neuropathy, foot wounds, peripheral vascular disease, Charcot arthropathy or   peripheral foot surgical procedures). Ertapenem is a once daily parenteral β-lactam antimicrobial agent with excellent in vitro activity against gram-positive and gram-negative aerobic and anaerobic bacteria generally associated with community-acquired and mixed infections. Aim of the work was to study in vitro activity of Ertapenem against bacterial isolates from diabetic foot infection and comparing its effect with that of other antimicrobials used for treatment of these infections. Materials and methods; thirty diabetic patients with infected foot ulcer were enrolled in this study, they   were admitted to the department of surgery, Ain Shams University Hospital from November 2010 to April 2011. Swabs were collected from the infected ulcers, cultured areobically and anaerobically. The isolated microorganisms were identified by conventional microbiological methods. Antimicrobial susceptibility test were done on bacterial isolates using the disc diffusion method. Results ; Proteus vulgaris was the most common isolated organism from infected foot ulcers among studied cases as it was isolated from 15/30 (50%) of cases, Staphylococcus aureus was isolated from 12/30 (40%)of cases, E. coli and Enterobacter cloacae each was isolated from 6/30(20%) of cases then Pseudomonas aerugenosa and Klebsiella pneumoniae each was isolated from 3(10%) of cases. Only 3/30(10%) anaerobic microorganism were isolated (Peptostreptococcus). Mixed infection was detected in 17/30(56.7%) of cases. Ertapenem was effective against most isolated organisms (except Pseudomonas aeruginosa) with highly statistically significant difference between sensitivity and resistance.Conclution; Ertapenem has a good activity against most isolates from infected diabetic foot ulcer.

4/29 IMPACT OF MYCOPLASMA HOMINIS AND UREAPLASMA UREALYTICUM CERVICAL COLONIZATION ON ICSI OUTCOME.

Ibrahim Ghanem, Hassan Mansour Hegab,Nesrine Fathi Hanafy* Aly Youssef

Department of Obstetrics and Gynecology, *Medical Microbiology and Immunology, Faculty of Medicine, Alexandria University

Contamination of cell lines with mycoplasma is one of the major problems in cell culturing, including embryo cultures. It may lead to deterioration in media quality and extensive affection of cultured cells. Mycoplasma hominis and Ureaplasma urealyticum, as vaginal colonizer might contaminate oocytes being retrieved per vaginum and affect embryo yield in their culture, during ICSI procedure. In this study, we aim at evaluation of impact of cervico-vaginal colonization with Mycoplasma hominis (M.h) & Ureaplasma urealyticum (U.u) at time of oocyte retrieval on ICSI outcome. Cervico-vaginal swabs have been collected from a cohort of 40 infertile females undergoing ICSI, and from a control group of multiparus women. All samples have been subjected to detection via PCR amplification for Mycoplasma hominis and Ureaplasma urealyticum using specific primers. M.h was revealed in 9 (22.5 %) of the cases and one (5 %) of controls while U.u. was revealed in 14 (35%) of cases & 8 (40%) of controls. Our results revealed that Mycoplasma hominis colonization   was found to affect the oocyte yield and pregnancy in ICSI cycles. While no significant impact on ICSI outcome have been noted with Ureaplasma ureatlyticum colonization.

5/29 ACINETOBACTER BAUMANNII ISOLATES FROM DIFFERENT EGYPTIANINTENSIVE CARE UNITS: PHENOTYPIC METHODS FOR DETECTION OF POTENTIAL RESISTANCE MECHANISMS TO CEPHALOSPORINS AND CARBAPENEMS

Moslhey S. Mansy*, Hamido M. Hefni*, Wael M. Tawakol and

Mohamed F. El-Badawy

Department Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology, 6th October, Egypt.

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

Acinetobacter baumanii (A. baumanii) has emerged as important organism inside intensive care unit (ICU) settings. Metallo-beta-lactamases (MBLS), Amp C- beta-lactamases and carbapenemase mediated resistance is an emerging threat in hospital isolates of A.baumannii. The goal of this research is to investigate the potential resistance mechanism for cephalosporins and carbapenems. Thirty one isolates from sputum, endotracheal tube (ETT), central venous catheter CVC, blood, urinary catheter, pus, surgical drain and surgical wound were recovered from 31 different cases hospitalized in different Egyptian ICUs. All specimens were subjected to isolation and identification using selective media and different biochemical tests. A. baumannii isolates were confirmed and identified to species level using API 10S. Antibiogram was done for all isolates using different antibiotic discs by Kirby Bauer disc diffusion method. Meropenem resistant isolates were subjected to modified Hodge test (MHT), ethylene diamine tetra acetic acid (EDTA) disc synergy test (EDS) and Amp C disc test to detect the production of carbapenemase, MBL and Amp C respectively. This study revealed that 100% were MBL producers by EDS, 52% were Amp C producers by Amp C disc test and none of them were carbapenemase producers by MHT. This study demonstrated that EDS test seems to be a better method for detection of resistance to meropenem than MHT.

6/29 PREVALENCE OF MICROBIAL OCULAR INFECTIONS AMONG PATIENTS ATTENDING SOME EGYPTIAN HOSPITALS

Moselhy S. Mansy, Hamido M. Hefni and Mohamed A. Ghonim

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

       Two hundred ninety six patients with ocular infections were subjected to clinical and microbiological investigations. They were selected from patients attending the outpatient clinic of AL-Hussein University Hospital and Sayed Galal University Hospitals (Cairo governate) in addition to Tanta University Hospital for ophthalmology. Microbiological methods for isolation, identification and antimicrobial susceptibility testing were used. The most frequently clinical feature was conjunctivitis (46.3%). Our finding showed that the prevalence of bacterial infection in the ocular samples was (87.8%). Staphylococcus aureus was the most common causative agent, being responsible for (31.2%) of the all cases, it was followed by Staphylococcus epidermidis (24.2%) and Klebsiella pneumonia (20%). The frequency of bacterial ocular infection in children aged 2-<14 years was higher than other age groups. The flouroquinolones especially moxifloxacin and third generation cephalosporins were highly effective against all bacterial isolates. On other hand, most bacterial isolates were resistant to penicillin, ampecillin, tetracycline and chloramephenicol.

7/29 NOSOCOMIAL INFECTIONS: BCTERIAL PREVALENCEAND ANTIBIOTIC RESISTANCE PATTERN AMONG ISOLATES RECOVERD FROM DIFFERENT EGYPTIAN INTENSIVE CARE UNITS

Hamido M. Hefni*, Moslhey S. Mansy*, Wael M. Tawakol and

Mohamed F. El-Badawy

     Department Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology, 6th October, Egypt.

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

Nosocomial infections are one of the most frequent medical complications affecting patients   admitted to the intensive care unit (ICU). The flourishing of ICU with the prolonged stays and close proximity of many patients whose physical and immunologic defenses are compromised has created settings in which the emergence and spread of resistant bacteria have accelerated. The goal of this research is to investigate the prevalence and antimicrobial spectrum of Gram positive and Gram negative bacteria affecting ICU patients. In this study a total of two hundred and fifty-five specimens as sputum, endotracheal tube (ETT) , central venous catheter (CVC) ,central venous pressure (CVP), nasogastric tube (NGT),cerebrospinal fluid (CSF), eye ,nasal, urine , blood, pus , surgical drain , wound and bedsore which recovered from 198 different cases. All specimens were subjected to isolation identification using different media and biochemical tests. Antibiogram was done for all bacterial isolates using Kirby Bauer disc diffusion method. Microbiological identification revealed that 30.58% of isolates were Gram positive and 69.42% were Gram negative. Coagulase negative staphylococci were the most prevalent isolates (16.86%) among Gram positive isolates, while E.coli were the most prevalent isolate (15.69%) among Gram negative isolates. Antibiogram revealed that vancomycin (97.43%) was the most effective antibiotic against Gram positive followed by Amikacin 69.23%; on the other hand Meropenem 74.01% was the effective among Gram negative isolates followed by Amikacin 66.10%. The results of this study demonstrate a relative high resistance to antimicrobial among ICU bacterial isolatesdue to indiscriminate use of antibiotics inside these units.

8/29 BIOINFORMATICS/BIOTECHNOLOGICAL MODEL FOR THE PRODUCTION OF POLYHYDROXYBUTYRATE FROM MIXED BACILLUS CULTURE

Soheir R. Salem and Amro A. Amara*

Faculty of Education, Biological and Geological Science Department, Alexandria University, Alexandria, Egypt.

*Protein Research Department, Genetic Engineering and Biotechnology Research Institute for Scientific Research and Technological Applications, Borg el Arab, Egypt

Bacillus was the first species reported to produce polyhydroxybutyrate (PHB). The use of mixed culture for the production of biotechnological products has many advantages over using a single culture due to the ability of using heterogeneous substrates. In this study the PhaC synthases of four Bacillus species have been extensively screened using different genomic and proteomic database. The Bacillus species represent: Bacillus subtilis, Bacillus pumilus, Bacillus thuringienesis and Bacillus licheniformis. The aim of this study is to build a bioinformatics/biotechnological model for analysis the possibility of production of certain products (e.g. PHB) using mixed bacterial culture. One PhaC synthase represent B. thuringiensis has been found in protein Blast search and the other three sequences have been extracted from complete genomic sequences of the other three strains. For proving our hypothesis in vivo experiments have been conducted using homogeneous and heterogeneous media constituents. Equal inoculums have been used from each strain using simple method. The different produced PHB have been converted to crotonic acid and determined spectrophotometrically and a modified method has been used. Four media have been evaluated regarding to the ability of the four used Bacillus to produce PHB. Even using mixed culture of Bacillus; the production processes proved to be sensitive to changes in media compositions. Amounts of the PHB produced in this study were 1.9; 0.7; 0.6 and 2.7 g/l/24hr for each experiment and its medium respectively. The differences between the overall media constituents of the four experiments are highly significant p<0.001. Out of the eleven used media constituents, glucose and trace elements were highly significant p<0.001. Cottonseed oil, skim milk and FeSO4 were significant P<0.002, 0.004, 0.004 respectively. This simple method can be used as preliminary method for quick evaluation for different media compositions in case of using mixed culture. We recommended using of both bioinformatics and biotechnological tools to find suitable models for the different production processes in biotechnology.

9/29 VANCOMYCIN RESISTANT ENTEROCOCCI COLONIZING THE INTESTINAL TRACTS OF HOSPITALIZED PATIENTS

Hend Alsayed Sharaf

MedicalMicrobiology & Immunology Department, Faculty of Medicine,

Zagazig University

Enterococci are important not only because they are a leading cause of nosocomial infections, but also because they have a significant role in dissemination and persistence of antimicrobial resistance. Objectives: This study aimed to determine the distribution of vancomycin-resistant enterococci (VRE) in surgery and hematology Departments, Zagazig University Hospitals, study some factors that may increase the risk of VRE colonization and determine the phenotype/genotype relation of isolated VRE. Methods: This study was conducted from September 2006 to June 2007, at Medical Microbiology and Immunology Department, Zagazig University.Fecal samples were taken from 200 patients (100 from each hematology and surgery wards). Each sample was screened for the presence of VRE and for each isolate Etest to determine MIC and Multiplex PCR to detect vanA or vanB genes were done. A case-control study was performed to study some risk factors of VRE colonization. Results: Thirteen (6.5%) VRE were isolated out of 200 stool samples. Four VRE (4%) were isolated from the surgical ward patients and 9 (9%) VRE were isolated from the hematology ward patients. PCR genotyping results were consistent with the resistance phenotype for all except one VRE isolate; that had vanB phenotype and vanA genotype, as it was teicoplanin susceptible in spite of vanA gene. By studying risk factors of colonization by VRE, we found that patients stayed more than 30 days in the hospital were significantly associated with VRE carriage (P=0.004). Also previous receiving of glycopeptides was significantly associated with VRE colonization in both Departments (P=0.001). Conclusion:Antimicrobial resistance is an increasing problem and challengeworldwide. The emergence of enterococci resistant to glycopeptides poses the particular dilemma of controlling transmission of these organisms from patient to patient. Although the manner of colonization with VRE remains unclear, rapid identification of VRE-colonized patients by means of surveillance cultures and MIC determination are often required.

10/29 STUDY OF RESPIRATORY SYNCYTIAL VIRUS INFECTIONS AMONG CHILDREN WITH ACUTE RESPIRATORY SYMPTOMS

Hend Alsayed Sharaf, MahaKamal Gohar and Randa Saddek

MedicalMicrobiology & Immunology Department,Faculty of Medicine,

Zagazig University

Background and objectives: Respiratory syncytial virus (RSV) is a major cause of serious lower respiratory tract illness in infants and young children worldwide. This study aimed to determine the distribution of subgroup A and B of RSV in hospitalized infants and young children, study some factors that may increase the risk of RSV infections and to compare between tissue culture and multiplex Reverse Transcriptase Polymerase Chain Reaction (mRT-PCR) in diagnosis of RSV. Methods: This study was conducted from October 2009 to April 2010, at Medical Microbiology and Immunology Department, Faculty of Medicine, Zagazig University. A total of 100 nasopharyngeal swabs collected from infants and young children with acute lower respiratory tract infections during one epidemic period in Zagazig University Hospitals were subjected to viral isolation by tissue culture. RSV cytopathic effects (CPE) were confirmed as RSV by immunoflourescence. Also all samples were subjected to mRT-PCR to determine subgroup A and B of RSV. Results: Twenty three samples out of 100 (23%) were confirmed culture positive while 38 (38%) were positive using mRT-PCR. The present data confirm the agreement of viral culture and mRT-PCR for diagnosis of RSV. Strains of subgroup A accounted for 65.78% of cases, while type B strains were 34.21%. By studying different groups of patients, we found that age group from 0 to 6 months was more suspected to RSV A and B infections (P=0.001 and 0.01) respectively. Also being a male was statistically significant in both type A and B RSV infected patients (P= 0.02 and 0.049) respectively. Both types A and B, RSV positive cases were significantly high in those using artificial feeding (P=0.009 and 0.01 respectively). Both types of RSV were high in cold season and reached a peak in January. Conclusion: RSV was found as a major problem facing our community, especially during cold seasons. Future planning for vaccine development requires further understanding of the genetic composition of the RSV strains prevalent in the target population. Multiplex RT-PCR was a suitable method for diagnosis of RSV infections and an effective tool for characterization of RSV circulation patterns in communities.

11/29 IN VITRO ACTIVITY OF TIGECYCLINE AGAINST CLINICAL ISOLATES OF ESBLS PRODUCING GRAM NEGATIVE BACILLI

Sanaa M. Ibrahim, Omima Sayed, Lamiaa A. Adel, Hamed H. Abostate*

Medical Microbiology & Immunology Department, and *General surgery Department, Faculty of Medicine, Ain Shams University

Multi-drug resistant organisms (MDRO) represent a difficult challenge due to the limited therapeutic options. Extended- spectrum beta- lactamase (ESBL) producing Gram negative bacilli are important example for these MDRO that are resistant to most commonly used antimicrobials. The multidrug-resistant properties of the ESBLs limit therapeutic options, and there is an urgent need for new agents that are able to combat these pathogens. The aim of the study was to evaluate the in vitro activity of tigecycline against ESBL producing clinical isolates of Gram negative bacilli. Samples from different types of infections were collected in the period from Marchto October2010 and have been examined for isolation and identification of the causative microorganism by using conventional microbiological methods. The BBL crystal identification system was used to confirm identification of Gram negative bacilli. Thirty ESBL- strains were detected by disc diffusion test using ceftazidime, ceftazidime - clavulanate. Confirmation for ESBL production was done by E- test. These strains will be subjected to the tigecycline susceptibility testing using disc diffusion method and tigecycline MIC using the E-test strips the MIC breakpoint for susceptibility will be taken as ≤ 2 μg/ml for Gram- negative bacteria. The result of the study showed that, Frequency of ESBL production among each of the isolated gram negative bacilli by confirmatory methods was that K.pneumoniae was the species with the highest frequency (12/23, 52.2 %) followed by P.mirabilis (2/4, 50%), and then E.coli (7/21, 33.4 %). For P. aeruginosa (7/14, 50%) were ESBL positive.  For Acinetobacter baumannii (2/7, 28.6%) were ESBL positive. All the isolates of E.coli and K. pneumoniae were sensitive to tigecycline by disc diffusion method and E-test. However, only 14.3% of P.aeruginosa, 50% of Acintobacter spp, and 50% of P.mirabilis were sensitive to tigecycline.Conclusion. Tigecycline represents a significant step forward over the older semisynthetic tetracyclines, showing excellent in vitro activity against strains for which adequate therapy has been limited.

12/29 CHARACTERIZATION OF SOLUBLE ENZYME II (EII) MANNITOL OF PHOSPHOENOLPYRUVATE DEPENDENT PHOSPHOTRANSFERASE SYSTEM (PTS) OF ESCHERICHIA COLI

Mohammad M. Aboulwafa and Nayera A. Moneib*

Department of Microbiology & Immunology, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt

*Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, King Abdulaziz University, Jeddah, 21589, Kingdom of Saudi Arabia

In this study, it was found that the 2 h high speed supernatant (2 HSS) of crude cell lysate of E. coli exhibits enzyme II mannitol activity when assayed for both phosphoenolpyruvate-dependent (PEP) or transphosphorylation (TP) activities. Gel filtration of this 2 HSS resulted in the appearance of two PEP and only one TP activity peak. When 2 h HSS was separately treated with different concentrations of urea, n-butanol and Na deoxycholate it was observed that: (1) treatment with urea (4 M and 8 M) and Na deoxycholate (0.2 and 0.4%) has a little effect on the PEP activity with somewhat appreciable enhancing effect in case of Na deoxycholate treatment. The transphosphorylation (TP) activity exhibited an appreciable increase relative to control upon urea and Na deoxycholate treatment and the effect with Na deoxycholate was more than that with urea. In contrast, treatment with n-butanol (4, 5.3, 6.7 and 8%) reduced both PEP and TP activities and this effect was proportional to the concentration applied. Combined treatment with butanol and Na deoxycholate but not with butanol and urea sharply decreased both PEP and TP activities. Urea imparted a protective action for the adverse effect of butanol. While untreated 2 h HSS gives two activity peaks (1 & 2) for PEP reaction and one peak for TP reaction, the low speed supernatant of butanol treated 2 h HSS showed only one activity peak for PEP reaction (peak 2) and disappearance of peak 1 for both PEP and TP reaction. The results show clear evidence for: (i) the different sensitivities of both PEP and TP activities of enzyme II mannitol for urea, n butanol and Na deoxycholate, (ii) the presence of two physical forms of EII mannitol in the 2 HSS with different sensitivities for butanol.

13/29 EFFECTS OF GAMMA AND ELECTRON BEAM IRRADIATION ON VIABILITY AND AMPLIFIABLE DNA ELIMINATION OF STAPHYLOCOCCUS AUREUS

Mona M. K. Shehata, Fatma Alzahraa M.Gomaa* and Zeinab H.Helal*

Drug Radiation Research Department,National Center for Radiation Research and Technology, Atomic Energy Authority, P.O. Box 29, Nasr City, Cairo, Egypt.

*Microbiology and Immunology Department, Faculty of Pharmacy,

Al-Azhar University, Nasr City, Cairo, Egypt.

Owing to the ability of real-time PCR to detect DNA released from dead cells, a risk exists, that DNA from dead cells can result in false-positive PCR signals. In this study, gamma and electron beam irradiation are compared for their effects, on elimination of amplifiable DNA which is an issue of relevance to molecular PCR applications, as an effective decontamination method. The effects of both the methods on the viability of Staphylococcus aureus using quantitative cultures were evaluated. Quantitative real-time PCR based assay of 16S rRNA gene fragments amplified from Staphylococcus aureus were performed. Differences in radiation sensitivities of genomic DNA extracted from bacteria before subjecting to irradiation in comparison with DNA residing within viable bacterial cells at the time of irradiation were also evaluated. Viability was abrogated at 2.6kGy and 3kGy and the radiation dose required to produce 1–log10 or 90% reduction in viable microorganisms (D10- values) was 0.32 kGy& 1.73kGy for γ- and electron beam irradiation, respectively. This study showed that gamma irradiation could eliminate amplifiable DNA at the tested doses (12-25kGy) compared to accelerated electrons which has negligible reduction in quantity with increasing the dose. D10- values for amplifiable DNA were significantly higher for DNA extracted from viable bacterial cells irradiated with electron beam than that with gamma radiation (32.17 & 3.99kGy respectively; p=0). These attributes are important in clinical practice with increasing use of molecular amplification techniques in microbiological diagnostic applications.

14/29 PHENOTYPIC AND GENOTYPIC ANALYSIS OF BIOFILM FORMING STAPHYLOCOCCI

Gamal Fadl Mahmoud Gad, Hamada Mohamed Mohamed* and

Tharwat Ragheb El khamisy*

Microbiology Department, Faculty of Pharmacy, El-Minia University,

El-Minia, Egypt

*Microbiology Department, Faculty of Pharmacy, Al Azhar University, Assuit, Egypt

            Coagulase positive Staphylococci (COPS) and Coagulase negative Staphylococci (CONS) are responsible for many of biofilm-related infections. They are common causes of nosocomial infections. A total of 180 clinical strains of Staphylococcal spp were isolated from different specimens. Strains were screened for biofilm formation by Tube method (TM), Congo red agar (CRA) method, and by tissue culture plate (TCP) method. It was found that COPS and CONS isolated from urine specimens were more able to produce biofilms (85.7 and 79.3 % respectively) than those isolated from other specimens. Also, it was found that CONS had the ability to produce biofilm by the three methods (CRA, ST and TCP) more than COPS. Effects of physiological factors (glucose 1%, NaCl 4% and different pH values) were studied. Addition of glucose (1%) enhanced biofilm formation while addition of NaCl (4%) decreased the formation for both COPS and CONS. Different values of pHs highly affected the production of biofilm. The results revealed that pH 7.2 was an optimum for production of biofilm while higher values highlydecreased biofilm production. The presence of icaA and icaD genes responsible for biofilm formation was detected by PCR technique and the presence of bap (biofilm-associated protein) is detected by poly acryl amide gel electrophoresis. Of the 180 Staphylococcal strains,114 were coagulase positive and 66 were coagulase negative. Screening by Congo red agar and tube method revealed that 50 (43.8%) and 65 (57%) were biofilm producer for COPS respectively, while 39 (59%) and 27 (41%) were biofilm producers for CONS. Screening by TCP revealed that out of the 114 coagulase positive strains, 73 (64%) were biofilm producers and 45 (68.2%) out of the 66 coagulase negative strains were biofilm producers. Detection of icaA and icaD genes agreed with TCP results. As a conclusion, the TCP method was found to be sensitive, accurate and has the advantage of being a quantitative model to study the adherence of staphylococci on biomedical devices and agree with the results of PCR

15/29 cELLULASE-fREE xYLANASE pRODUCTION BY TWO RADIATED STRAINS OF THERMOMYCES LANUGINOSUS ISOLATED FROM EGYPT AND YEMEN SOILS USING SOLID STATE FERMENTATION

Loutfy A.A. Moussa*, Khairyia A.Yousef**,Usama F. Ali, Zeinab M.   Ibrahim and George S. Isaac

Department of Biological sciences, Faculty of Education, Ain Shams University.

* Department of Microbiology, Faculty of Science, Ain Shams University.

**Department of Microbiology, National Center of Radiation Research and Technology (NCRRT).

The potentiality of Thermomyces lanuginosus A72 and T. lanuginosus YMN72 were screened on five natural lignocellulosic substrates for cellulase-free xylanase production. Highest xylanase production showed on cane bagasse and corn cobs when added to growth medium in the ratio (1:3, w/v) and moisture content 77.14 and 79.52% for T. lanuginosus A72 and T. lanuginosus YMN72, respectively. Maximum xylanase activity reached at 6 and 5 old actively growing cultures (6.0%, v/v) for T. lanuginosus A72 and T. lanuginosus YMN72; respectively. Addition of glycerol (1.5%, v/v) and beef extract (0.1%, w/v) to the growth medium of T. lanuginosus A72 gave 1.26 fold increases in the xylanase activity compared with the control value. While, addition of tween 80 (2.0%, v/v) and malt extract (0.15%, w/v) to the growth medium of T. lanuginosus YMN72 gave 1.18 fold increase in the xylanase activity compared with the control value. The maximum xylanase production reached at dose 0.5 kGy of gamma radiation giving 39522 and 43882 U/g with increasing 102 and 105% as compared with the control value of T. lanuginosus A72 and T. lanuginosus YMN72, respectively.

16/29 ENCOURAGEMENT OF CHITINASE PRODUCTION FROM ASPERGILLUS NIGER AND TALAROMYCES THERMOPHILUS BY RADIATION USING CRAB SHELLFISH WASTE AS A SUBSTRATE IN SOLID STATE CULTIVATION

Loutfy A.A. Moussa*, Khairyia A.Yousef**, Usama F. Ali, Zeinab M. Ibrahim

and Omayma M. El-Mahdi*

Department of Biological sciences, Faculty of Education, Ain Shams University.

*Department of Microbiology, Faculty of Science, Ain Shams University.

**Department of Microbiology, National Center of Radiation Research and Technology (NCRRT).

In solid state fermentation (SSF) Aspergillus niger and Talaromyces thermophilus were screened on four substrates for chitinase production. A. niger gave the maximum production (15.564 mU/g) when beet molasses was used as the basal solution with crab shell chitin waste in the ratio of 1:2 (Moisture content; MC, 69.37%). T. thermophilus gave the maximum production (10.960 mU/g) when cane molasses was used as the basal solution with crab shell chitin waste in the ratio of 1:1 (Moisture content; MC, 55.45%). Maximum chitinase production was reached at 6 and 7 days for A. niger and T. thermophilus respectively. The maximum chitinase production was obtained after exposing the fungal slants of A. niger and T. thermophilus to dose level 0.5 KGy of gamma radiation giving 17.564 and 11.994 mU/g with increasing 163.158 and 109.943% as compared with the control value of A. niger and T. thermophilus respectively.


17/29 EFFECT OF VARIOUS ANTIBIOTICS ON PLANKTONIC AND BIOFILM PRODUCING STAPHYLOCOCCI

Gamal F.M. Gad and Hamada M. Mohamed*

Microbiology Department, Faculty of Pharmacy, El-Minia University,

El-Minia, Egypt

*Microbiology Department, Faculty of Pharmacy, Al-Azhar University, Assuit, Egypt

The Pathogenesis of Staphylococcal speciesis attributed to a combination of extra-cellularfactors and other properties such as adherence andbiofilm formation. The nature of biofilm structure and the physiological attributesof biofilm organisms confer an inherent resistance to antimicrobialagents and are in need for more studies. In this study, 180 strains of Staphylococcal spp were isolated from 415 clinical samples collected from patients attending Assiut University Hospitals. The strains were identified microscopically, by biochemical methods and API strips for Staph. Out of the 180 Staphylococcal spp, 114 were coagulase positive and 66 were coagulase negative. All strains were screened for biofilm production by tissue culture plate (TCP). Screening by TCP, for detection of biofilm, revealed that out of the 114 coagulase positive staphylococci (COPS), 73 (64%) were biofilm producers. For the 66 coagulase negative staphylococci (CONS), 45 (68.2%) were biofilm producers. The strains that formed biofilm (by tissue culture plate method) were screened by scanning electron microscope (SEM) to study their ability to produce biofilm and to study effect of certain antibiotics on biofilm formation. Results revealed that biofilm forming strains (by TCP) were positive for biofilm production through SEM. Also, scanning electron microscope showed that antibiotics reduced biofilm formation. Resistance of planktonic and biofilm forming cells to antibiotics were studied using disc diffusion method. Biofilm forming cells were more resistant to antibiotics than planktonic cells. MICs of antibiotics affecting planktonic and Biofilm cells were determined using microbroth dilution method and a new biotimer assay method. It was found that, MICs of biofilm cells were higher than that of Planktonic cells (either by microbroth dilution method or biotimer assay method).Also, it was found that MICs by microbroth dilution method agree with that of biotimer assay method. The MICs by biotimer assay method, as a new method, was found to be sensitive, accurate, reliable and agree with the results broth dilution method and SEM.

18/29 MICROBIOLOGICAL STUDY OF DIABETIC FOOT INFECTION

Essam Abou-El-yazed, Ibrahim M.Alhosiny* and Ihab A Alfakhrany*.

Department of Microbiology and Immunology, Faculty of Medicine, Al- Azhar University and *Vascular Surgery Department, Faculty of Medicine for Girls, Al- Azhar University, Cairo, Egypt

This study was conducted on 100 patients presented by diabetic foot infections(DFIS) admitted to Vascular Unit of Surgery at Al- Zhraa   University Hospital during the periods from January 2008-to November 2009, their age ranged from45-78years (59.2±5.1),45 Patients are males, and 55 patients are females, Pus aspirates from the abscesses and swabs were taken on transport media and inoculated on nutrient agar, blood agar, Mac Conkey’s agar, Robertson's cooked meat media and Sabouraud’s agar culture media under aerobic and anaerobic conditions. In addition, the specimens from patients with cellulitis and gangrene were inoculated into blood agar for anaerobic culture. After incubation for 24-48 hours at 370 C, any detected growths were examined by colonial morphology and the effect of organisms on different culture media. Gram stained film, and biochemical reactions were done for full identification of any growth. Escherichia coli was the most predominant organisms (40%), followed by Staphylococcus aureus (30%), Candida albicans (20%), Proteus vulgaries, Micrococcus (15%) each, Staphylococcus epidermidis (10%), Klebsiella pneumoniae   (5 %),   Bacteroides fragilis (4%), Pseudomonas aeruginosa (3%), Clostridium perfringens (2%) and Clostridium teteni (1%) respectively. The patients were treated with antibacterial agents according to culture and antibacterial susceptibility pattern. Dressings were done twice daily, surgical interventions were done wherever needed for cellulitis, and dressing and depridridment for extensive infections and amputation were done for patients with gangrene.

19/29 CHARACTERIZATION AND ANTIMICROBIAL SUSCEPTIBILITY OF STAPHYLOCOCCUS SPECIES ISOLATED FROM CANCER PATIENTS ON CHEMOTHERAPY IN EGYPT

El Domany R., S. Abdelatif* and N. A. Abdelaziz**

Department of Microbiology, Faculty of Pharmacy Helwan and Future** Universities, National Cancer Institute, Egypt*

Compromised immune system of cancer patients makes them an easy target for bacterial infection. Recently, several reports indicated an increasing incidence of Staphylococcus isolated from cancer patients. The presented study analyzed the frequency of Staphylococcus species isolated from different infection sites, full characterization of isolated microorganisms, quantitative and qualitative determination of antimicrobial susceptibility patterns of Staphylococcus species isolated from cancer patients, to many important antimicrobial agents using Microscan PID , Microscan WalkAway Systems and manual methods The results showed that The main isolated Staphylococcus from sputum , throat and pus swabs were S.aureus (38.6%) ,(46.7%) and (100%) respectively .The main isolated Staphylococcus from all specimens were(CNS 62.8%; followed by S.aureus 37.2%).S.aureus isolates were highly susceptible to Linezolid and Sufamethoxazole/trimethoprim followed by Synercid(quinupristin/dalfopristin) and Vancomycin .They were still susceptible to few other antimicrobial agents such as Gatifloxacin , Rifampin and Moxifloxacin(31.2% resistance).However, they exhibited higher resistance to most other antimicrobial tested. Similarly, CNS isolates exhibited 42.6% resistance to Linezolid and 38.9% resistance to Vancomycin. They were still susceptible to few other antimicrobial agents such as Moxifloxacin(38.9% resistance) and Rifampin(48.1% resistance).However, they exhibited higher resistance to most other antimicrobial tested. Almost all of staphylococcal isolates tested were resistant to oxacillin.


20/29 METALLO BETA LACTAMASE PRODUCTION AMONG CARBAPENEM NON-SUSCEPTIBLE PSEUDOMONAS AERUGINOSACLINICAL ISOLATES

Nadia H. Ouda and Iman E. Wali

Department of Medical Microbiology & Immunology,Faculty of Medicine,

Cairo University, Egypt

Metallo beta lactamase (MBL) enzymes play a critical role in carbapenem non-susceptible Pseudomonas aeruginosa (CNPA), with risk of hospital spread and fear of limited therapeutic options. There are no standardized guidelines for detection of these enzymes. Our objectives were to determine the frequency of MBL production phenotypically among clinical CNPA isolates and to compare the performance of the applied detection methods. In addition, we aimed to determine which carbapenem would be more reliable in preliminary screening for MBL producing CNPA and to evaluate available therapeutic options. A total of 228 non-duplicate clinical isolates of Pseudomonas aeruginosa were tested for their non-susceptibility to imipenem, meropenem or doripenem by disk diffusion method. Ninety CNPA isolates were tested for MBL production by the double-disk synergy test (DDST), combined-disk test (CDT) and MBL E-test methods, as well as for their susceptibility to different antibiotics other than the carbapenems. MBL E-test detected 66/90 (73.3%) MBL producing CNPA; whereas, the DDST and CDT detected MBL production in 63/90 (70%) and 57/90 (63.3%) respectively. Considering E-test as the gold standard, sensitivity and specificity of DDST and CDT were 95.5%, 100% and 86.4%, 100% respectively, which was statistically significant. MBL producers were highly resistant to commonly used antibiotics. However, they were highly susceptible to colistin (100%) and fosfomycin (81.8%). In conclusion, 73.3% of CNPA were MBL producers by E-test. Imipenem was the most reliable carbapenem for screening MBL producers. The use of DDST or CDT proved equally effective for the detection of MBL production. Colistin and fosfomycin may be useful for the treatment of CNPA infections.

21/29 SOLID STATE FERMENTATION FOR THE PRODUCTION OF PENICILLIN V BY PENICILLIUM CHRYSOGENUM NRRL 824

El-Sayed E. Habib, Mona I. Shaaban, Mohamed A. El-Sokkary

and Wael A. El-Naggar

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

Different substrates corn steep solid, rice bran, rice straw, sugar cane bagasse and wheat bran were tested for the production of penicillin V from Penicillium chrysogenum NRRL 824 by solid state fermentation (SSF). Wheat bran served as the best substrate for penicillin V production. The effect of initial moisture content, particle size, inoculum and addition of nutrients was also estimated for enhanced penicillin V yield from wheat bran substrate. The maximum amount of penicillin V (142.5 µg/gm of substrate) was obtained at the 3rd days of fermentation when the initial moisture content was 70% and average particle size was 1.0–2.0 mm. In addition, highest penicillin V yield was obtained upon inoculating with 1 ml spore suspension (1x108 spores/ml) rather than inoculating with mycelia. However, Addition of nutrients to wheat bran decreased penicillin V yield. Furthermore, mixing wheat bran with different concentrations of glucose or lactose was associated with higher penicillin V yield in SSF in contrast to submerged fermentation indicating the highest ability of solid substrate to tolerate catabolite repression. Utilization of glucose or lactose (4%) in SSF provided the highly penicillin V yield (233.5 and 354 µg/gm of substrate, respectively) at the 4th day of fermentation. Comparison between penicillin V produced by submerged and SSF revealed the significance of wheat barn as substrate for penicillin V production in SSF.

22/29 INDUCTION TEST PHENOTYPES AMONG CLINICAL ISOLATES OF ERYTHROMYCIN RESISTANT STAPHYLOCOCCUS AUREUS

Nadia H. Ouda

Department of Medical Microbiology & Immunology,Faculty of Medicine,

Cairo University, Egypt

Staphylococcus aureus (S. aureus) is an important cause of health care- and community-associated infections worldwide. Clindamycin (CLI) is one of the alternative agents used to treat S. aureus infections and accurate identification of CLI resistance is important to prevent therapeutic failure. Macrolide-lincosamide-streptograminB (MLSB) resistance can be constitutive (cMLSB) or inducible (iMLSB). Unfortunately, iMLSB resistance is not detected by standard susceptibility tests. This study aimed to determine the frequency of different phenotypes associated with MLSB resistance among erythromycin-resistant isolates of S. aureus (ERSA) by using an in vitro induction test (D-zone test) and to study therapeutic options for these resistant phenotypes. Non-duplicate clinical isolates of S. aureus were tested for their resistance to erythromycin (ERY), CLI and methicillin. ERSA isolates were then tested for MLSB resistance by the D-zone test as well as for their susceptibility to other antibiotics. Out of 181 isolates of staphylococci, 125 (69.1%) were identified as S. aureus. Eighty three S. aureus isolates (66.4%) were ERY- resistant, they included 78.3% MRSA and 21.7% MSSA. By disc diffusion method 49.4% were susceptible to CLI. Using the D-zone test, iMLSB resistance phenotypes were recognized in 26.5% (D,15.7% & D+,10.8%), while 50.6% had shown cMLSB resistance phenotypes (HD,4.8% & R,45.8%) and true CLI susceptibility (MSB phenotype) were found in 22.9% (D-Neg). Among MRSA, both iMLSB and cMLSB resistant isolates (30.8% and 56.9%) were significantly more prevalent when compared to MSB isolates (12.3%). All MLSB resistant phenotypes (100%) were susceptible to vancomycin and linezolid. In conclusion, the prevalence of iMLSB, cMLSB and MSB phenotypes of ERSA was 26.5%, 50.6% and 22.9% respectively and routine D-zone test of S. aureus isolates is strongly recommended to prevent treatment failure.


23/29 MICROBIOLOGICAL STUDIES ON RESISTANCE PATTERN OF ANTIMICROBIAL AGENTS AMONG MINERAL WATER, INFANTILE FOODS AND DAIRY PRODUCTS PATHOGENS

Mohamed.S.Eldin Ashour, Moselhy.S. Mansy*, Ola.A. Abdelrahman** and Mahmoud. A. Elfaky

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

Department of Microbiology and Immunology, Faculty of Pharmacy (Boys* and Gurls**), Al-Azhar University.

Antimicrobial susceptibility testing of bacteria isolated from mineral water, infantile foods and dairy products samples was done using 21 different antimicrobial agents commonly used in the Egyptian market. Staphylococcus aureus susceptibility testing revealed that cefoperazone and erythromycin showed 100% activity followed by oxytetracycline, doxycycline and vancomycin (88.89%), gentamycin and ciprofloxacin (77.78%), amikacin (66.67%), and chloramphinicol (55.56%). Staph. epidermidis sensitivity testing revealed that cephalexin, amoxicillin, cefoperazone, levofloxacin and ofloxacin showed the highest activity (100% and 90.9% the others), followed by ampicillin, ciprofloxacin and erythromycin (81.8% each), amoxicillin+clavulonic acid, ampicillin+sulbactam, gentamycin, cefotaxim and rifampicin (72.72% each), cephapirin, cepharidin, amikacin, gatifloxacin and vancomycin (63.6% each). E.coli susceptibility testing revealed that Ampicillin+Sulbactam and gatifloxacin showed 100% activity, while amoxycillin+clavulonic acid and ofloxacin showed 91.67%, ampicillin, cefoperazone, gentamycin, levofloxacin and ciprofloxacin showed 83.33%, oxytetracycline and doxycyclin showed 75%, amikacin showed 66.67%. Cefoperazone, levofloxacin, ofloxacin and ofloxacin showed the best activity against Salmonella spp. (100%) followed by oxytetracycline and ciprofloxacin (87.5% each), doxycycline, amikacin and chloramphinicol (75% each).The antibiotic susceptibility testing of Enterobacter Spp. revealed that cefotaxime and cefoperazone showed 90% activity each, while cefapirin and vancomycin showed 80% each, cepharidin 70%.

 

24/29 COMPARISON OF QUANTIFERON-TB GOLD IN-TUBE TEST AND TUBERCULIN SKIN TEST FOR DETECTING LATENT TUBERCULOSIS INFECTION IN IMMUNOCOMPROMISED PATIENTS

Somaia Abd El-Latif Eissa, Zeinab Abd El-Khalek Ibraheim, Eman Ahmed El- Seadi, Rafat Mohamed Abd El-Fatah* and Eman Abd Raboh**

Department of Medical Microbiology & Immunology, Faculty of Medicine, Cairo University, *National Cancer Institute, **Department of Internal Medicine, Faculty of Medicine, Al-Azhar university

Interferon-γ release assay (IGRA) may improve the current level of diagnostic accuracy for latent tuberculosis infection (LTBI). The QuantiFERON-TB Gold – In tube (QFT-IT) has been used in selected populations and shows higher specificity than the tuberculin skin test (TST). The present study compared the performance of TST with that of QFT-IT for the diagnosis of LTBI in immunocompromised patients. A total of 210 subjects (117 immunocompromised, and 93 immunocompetent) were prospectively enrolled in the present study. All participants underwent TST and QFT-IT testing. Comparison of the results of the two tests showed that significantly fewer immunocompromised than immunocompetent patients had positive TST results (10.3% vs. 28%, p 0.001), whereas the percentage of positive QFT-IT results was comparable for both groups (21.4% vs. 25.5%). However, indeterminate QFT-IT results were more frequent in immunocompromised than immunocompetent patients (21.4% vs. 8.6%, p 0.021). Agreement between the TST and QFT-IT was fair for the immunocompromised group (κ= 0.38),but moderate agreement was observed for the immunocompetent group (κ = 0.57). Indeterminate QFT-IT results were associated with anaemia, lymphocytopenia, and hypoproteinemia.In conclusion; In immunocompromised patients, the QFT-IT may be more sensitive than the TST for detection of LTBI, however, the assay showed a considerable proportion of indeterminate results.Therefore, care must   be taken   in the interpretation of the QFT-IT test results for these patients.

25/29 METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS NASAL CARRIAGE AMONG NEWLY ADMITTED PATIENTS AT SOHAG UNIVERSITY HOSPITAL

Mona F. Mohamed, Abeer SH. Mohamed and Laila M. Yousef*

Medical Microbiology and Immunology Department, Clinical Pathology Department*; Faculty of Medicine, Sohag University

This study was conducted to detect nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and its antibiotic susceptibility pattern among newly admitted cases at Sohag University Hospital, from March to June 2010. Nasal swabs were taken from 420 newly admitted patients. The isolates were identified as Staphylococcus aureus based on morphology, Gram stain, coagulase and catalase tests, and mannitol salt agar fermentation. Antibiotic susceptibility pattern of isolated S. aureus was evaluated by disc diffusion method. All methicillin-resistant isolates were examined for the existence of the mecA gene by PCR technique. In total, 93 patients (22.1%) were found to be nasal carriers of S. aureus and all these strains were MRSA. A statistically significant difference was only found for antibiotics usage between those with and without MRSA colonization. Antibiotic susceptibility pattern of isolated S. aureus showed high susceptibility to linezolid, complete resistance to penicillin, oxacillin and cefoxitin and variable degrees of resistance to vancomycin, teicoplanin, amikacin, ciprofloxacin and tetracycline. At conclusion, S. aureus nasal colonization as well as methicillin- resistance among these isolates is common finding in our community.

26/29 MICROBIOLOGICAL STUDIES ON MINERAL WATER, INFANTILE FOODS AND DAIRY PRODUCTS RETAILED ON GREAT CAIRO MARKET

Mohamed S. Eldin Ashour, Moselhy S. Mansy*, Ola A. Abdelrahman** and Mahmoud A. Elfaky

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

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

In principle, almost all enteric and opportunistic pathogens that are transmissible by the faecal-oral route can spread by water and food as main reservoirs. A total of 284 different samples including 96 mineral bottled water samples, 72 infantile food samples and 116 dairy products samples collected from different areas in Cairo and examined for their microbiological quality. About ten percent (10.4%) of bottled water samples representing three companies (Schweppes, Dasani and Aqua) were contaminated with Enterobacter spp. Only non pathogenic bacteria, Bacillus subtilis and Staphylococcus epidermidis, and non pathogenic fungi (Aspergillus niger, A. fumigatus, Cladosporium spp. and Mucor spp.) were isolated from infantile food samples. Samples of all dairy products were subjected to qualitative tests for the detection of pathogenic microorganisms (S. aureus, E. coli, Salmonella spp., Shigella spp., Brucella spp., M. tuberculosis and C. albicans). Negative results were obtained with the exception of two cases. Dalla milk showed to be contaminated with S. aureus, E. coli, Salmonella spp., as well as Candida albicans.

27/29 BIOCHEMICAL CHARACTERISTICS OF CARBAPENEMASES EXTRACTED FROM PSEUDOMONAS AERUGINOSA RESISTANT TO CARBAPENEMS

El-Naggar, W., Abdelaziz, A.*, Zaki, E. M.**, Abd El Galil, K. H.; Shokralla, S.***; Barwa, R.and Khalaf, E.

Department of microbiology, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.

*Department of microbiology, Faculty of Pharmacy, Tanta University, Egypt.

**Department of clinical pathology, Faculty of Medicine, Mansoura University, Egypt.

***Biodiversity Institute of Ontario, University of Guelph, Guelph, ON N1G2W1, Canada.

Background: Carbapenems have the broadest antipseudomonal spectra of all β-lactams and play a fundamental role in the treatment of Pseudomonas aeruginosa infections. Resistance to carbapenems is due to interplay among different mechanisms. The aim of this study is to study the role of carbapenem-hydrolyzing enzymes on carbapenem resistance in Ps. aeruginosa and to determine the biochemical characteristics of these enzymes. Methodology: Two hundred isolates of Ps. aeruginosa were collected from Mansoura university hospitals; fifty three (53) isolates were resistant to imipenem and meropenem. They were studied for antibiotic susceptibility pattern, minimum inhibitory concentrations of both imipenem and meropenem, carbapenemases and AmpC enzymes were detected microbiologically in the resistant isolates. Biochemical properties of some extracted carbapenem-hydrolyzing enzymes were studied. Results: all carbapenem resistant strains were resistant to cefotaxime, ceftriaxone. The MICs of all tested strains were greater for imipenem than meropenem except strains no. 27 and 45. Twenty four strains (45.28%) were serine carbapenemases producers, while Forty one strains (77.35 %) were metallo-β-lactamases (MBL) producers using both double disk synergy test and combined disk test. Furthermore, forty four (83.01%) strains were AmpC producers. The majority of MBL were potent enzymes which diluted from 1:1 to 1:15 to give 50% of imipenem inactivation. pI of some selected β-lactamases were determined using isoelectrofocusing gel electrophoresis. They were at 8.5 indicating AmpC and at 6.2 which were common in all tested enzymes. Regarding Vmaxvalues, it was found that imipenem was more readily hydrolyzed than meropenem by all tested enzymes except enzyme No. 37. All tested MBL were inhibited by EDTA, while not all serine carbapenemases were inhibited by clavulanic acid. Conclusion: MBL production is an important mechanism of carbapenem resistance among Ps. aeruginosa. Carbapenemases other than MBL may also be responsible for carbapenem resistance. AmpC β-lactamases is also a contributory factor for carbapenem resistance among the isolates under study. All metalloenzymes were inhibited by EDTA but not affected by clavulanic acid, while not all serine carbapenemases were inhibited by clavulanic acid. From spectrophotometric studies, imipenem is better hydrolyzed by MBLs rather than meropenem, this agrees with MICs values for meropenem which are less than those of imipenem.

28/29 CHARACTERIZATION OF EXOPOLYSACCHARIDES (EPS) PRODUCED BY PSEUDOMONAS AND ITS EFFECT ON THE IMMUNE RESPONSE

Talaat R.Z., Ramadan M.O., Abdel Khalek H.S., El-Zahaby, H. A.*

Microbiology Department, Faculty of Medicine, Botany Department, Faculty of Science*, Tanta University, Egypt

Eleven strains of gram negative bacteria (Pseudomonas) were screened for EPS production 5 human and 6 plant pathogens. Three of eleven isolates screened, (Pseudomonas syringae pv. tomato,. davson and pv. coriandricola) were considered as highly exopolysaccharide producers. The tested bacteria which showed highest EPS production were subjected to different factors to test the influence of these factors on their growth and EPS production. The highest EPS production was obtained from selected strains when grown on king's B media at 30C for 3 days. Sucrose as a carbon source and peptone as a nitrogen source gave the highest EPS production. Ferrous sulphate increase the EPS production while magnesium sulphate decrease EPS production. EPS has an immunomodulating role as it enhances the immune response in immunized rabbit with purified EPS. It increases total and absolute neutrophillic counts, TNF-Alpha.

29/29 MAGNETICALLY MODIFIED MICROBIAL CELLS FOR EFFICIENT REMOVAL OF WATER SOLUBLE TEXTILE DYES

Ahmed I. El-Batal, Abdel-Gwad M. Hashem*, Yasser M. Ragab* and Ahmed I. Helal

National Center for Radiation Research and Technology, Cairo. Egypt. (NCRRT).

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

Low cost, locally available biomaterials were tested for their ability to remove some reactive textile dyes from aqueous solution. In this study, Aspergillus tamarii and Saccharomyces cerevisiae biomasses were subjected to different physical and chemical pretreatments and their combinations to obtain the best modification that could give the highest adsorption capacity of the textile dyes from water. Aspergillus tamarii and Saccharomyces cerevisiae cells were magnetically modified by treatment with ferrofluid (magnetite iron nanoparticle) solution in order to prepare a new type of magnetically responsive biocomposite material. The best treatment was a combination of autoclaving, treatment with 5% formaldehyde and initial pH 2 of dye solution. Both the Freundlich and Langmuir isotherm models were used to describe the adsorption patterns of the modified biomass.

30/29 MORPHOLOGICAL AND MOLECULAR IDENTIFICATION OF EGYPTIAN TRICHINELLA SPIRALS ISOLATES

Khaled Abd El-Aziz Mohammad and Alaa Abd El-Aziz Mohammad*

Parasitology Department, Faculty of Medicine (Damietta)and Faculty of Science (Cairo)*, Al-Azhar University

A microscopically and molecular biological study to identify and characterize the Egyptian Trichinella spiralsisolates patterns through application specific stain technique and multiplex polymerase chain reaction (Multiplex-PCR) was described. The application of Giemsa technique to stain compressed diaphragm and muscle samples obtained from rats experimentally infected with Trichinella spirals is described. Diaphragm samples from rats heavily infected with 20 muscle larvae per gram of body weight (20 ML/gbw) were cut into several pieces and stained with Giemsa, besides, muscle samples were stained with hematoxylin-eosin. The ML was observed under the microscope as blue structures surrounded by non-infected muscle cells, which appeared with a pink coloration; similar contrast was observed in both diaphragm pieces and muscle samples. Muscle larvae of all experimentally infected rats were analyzed by a multiplex polymerase chain reaction (Multiplex-PCR) to examine the patterns of isolates of Trichinellaspirals. The muscle larvae of Trichinella isolates showed a 385 bp band patterns.

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