Vol. 28, January, 2011.

1/28 QUORUM SENSING INHIBITORY ACTIVITY OF SPONGE DERIVED FUNGI FROM RED SEA, EGYPT

Amro Hanoraa,b

aMicrobiology and Immunology Department, Faculty of Pharmacy, Suez Canal University, Ismailia 41522, Egypt; bBiology Department, College of Arts and Science, Georgia State University, Atlanta, USA

Filamentous fungi produce a battery of secondary metabolites, some of which are already in clinical use as antimicrobial drugs. Fungi coexist with bacteria but lack active immune systems, so instead rely on chemical defense mechanisms. It was speculated that some of these secondary metabolites could interfere with bacterial QS communication. Ten marine derived fungi were chosen for this study. The crude extracts of Red Sea derived fungi shows QSI activity against QSIS2 system and Chromobacterium violaceum ATCC 12472 which response to (C4 and C12) and C6 homeserine lactones (AHLs) respectively. Crude extract of two fungal isolates 25 and 28 have strong QSI activity when tested with Chromobacterium violaceum ATCC 12472 compared to cinnamon and garlic. Co-culture of fungal isolate 25 with signal molecules (C4 and C12) enhances the QSI activity. Fractionation guided by QSI shows three fractions of crude extract of Aspergillus sp. having QSI activity. The study has important ecological and biotechnological implications in terms of microbial communication in natural environments and enhancement of secondary metabolite production.

 

 

2/28 ANTIMICROBIAL AND ANTIOXIDANT ACTIVITIES OF THE ESSENTIAL OIL AND VARIOUS EXTRACTS OF ROSEMARY, SAGE, THYME AND NIGELLA, AGAINST SALLMONELLA TYPHIMURIUM & LISTERIA MONOCYTOGENES AND THEIR ROLES ON LIPID STABILITY

Ahmed F. Abdel-Salam, Wael H. M El-Reffaei, Shahenda M. Elaby, Fatma A.A. Mostafa and Khaled M. Elsawy

Regional Center for Food and Feed (RCFF), Agric. Res. Center, Giza, Egypt.

Evaluation effect of some plants extract and their essential oils on microbial and oxidation activities in lipid source. In the current study the antimicrobial effect of different concentrations (1.25, 2.5, 5.0%) of essential oils and ethanoleic plant extracts rosemary, thyme, sage, and nigella on growth and survival of Salmonella typhimurium and Listeria monocytogenes in vitro and in deboned chicken meat breasts were evaluated. The different concentrations of essential oils and ethanoleic plant extracts of rosemary and thyme posses a weak antimicrobial activity against Salmonella typhimurium and Listeria monocytogenes in vitro, whereas both sage and nigella have been shown a stronger antimicrobial activity against Salmonella typhimurium and Listeria monocytogenes in deboned chicken breasts. The concentration 5.0% of essential oils and their ethanoleic plant extracts represented the optimum concentration for decreasing Salmonella typhimurium and Listeria monocytogenes density in liquid media and deboned chicken breasts. In addition, the results indicated that, the essential oils were preciously higher affect than ethanoleic plant extracts on growth and survival of Salmonella typhimurium and Listeria monocytogenes in liquid and deboned chicken breasts. The antioxidant activity have been performed by using technique of rancimat to elucidate Antioxidative Activity Index (AAI) and Induction period (IP) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging methods are more suitable posse’s methods to assess the antioxidants activity. The essential oils shown from nigella and follows by sage have been increase the AAI and IP at level 2.5 % EOs, except at level 5% the nigella EO showed higher antioxidant activity than sage. Their antioxidants activity was ranked in order to the activity against DPPH: rosemary>thyme> sage> nigella seed, as 89.21, 83.00, 80.61 and 52.50 %, respectively. Obviously that, the DPPH scavenging method for both of rosemary and thyme ethanoleic extract shown a higher antioxidative activity against DPPH than for nigella and sage ethanoleic extracts. The obtained results suggested that nigella, thyme, Sage and nigella seeds could be used as antimicrobial and antioxidants agents against food - borne bacteria and lipid oxidation, especially on their essential oils.

 

3/28 STUDY ON THE FLORA OF ASWAN BOTANICAL GARDEN

Ahmed M. Fawzy, Abd El-Halim A. Mohamed, Hafeez R. Habeeb

and Safwat A. Azer

Flora & Phyto-Taxonomy Researches Department, Horticultural Research

Institute, Agricultural Research Centre, Dokki, Eygpt.

The study on the flora of Aswan Botanical Garden recorded the presence of 65 species belong to 27 Angiospermae families. The percentages of represented families were: Gramineae by 18.5%, Leguminosae 15.5%, Compositae 9.3%, Euphorbiaceae 7.8%, and Convolvulaceae 6.3%, each of Chenopodiaceae, Cyperaceae, Malvaceae, Palmae, Salicaceae and Solanaceae were represented by 3.1%. While the percentage was 1.5% in each of Amaranthaceae, Boraginaceae, Ceratophyllaceae, Cruciferae, Mollunginaceae, Oxalidaceae, Plantagoniaceae, Polygonaceae, Portulacaceae, Primulaceae, Salvadoraceae, Tamaricaceae, Tiliaceae, Umbelliferae, Verbenaceae and Zygophyllaceae. The percentages of the annuals and perennials species in the studied area were 52.31 and 29.2 %; respectively, while the tress and shrubs were represented by 10.7 and 7.69 %; respectively. One annual parasite species was recorded: Cuscuta campestris Yunck. Two wild species: Salvadora persica L. and Cordia sinensis Lam. were cultivated in the studied area.

 

 

4/28 IMPACT OF INOCULATION WITH AZOSPIRILLUM BRASILENSE AND ZINC-ENRICHED AZOLLA AND OR MINERAL ZINC ON GROWTH AND PRODUCTIVITY OF RICE (ORYZA SATIVA L.)

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

Agricultural Microbiology Res. Department, Soils, Water and Environment Res. Inst., Giza, Egypt.

A pot experiment was carried out to evaluate microbial inoculation with Azospirillumbrasilense and two forms of zinc (Zinc-enriched azolla and ZnSO4.7 H2O) on yield and nutrients uptake by rice plants grown in clay soil. Inoculation with Azospirillumbrasilense at rats of 5 and 10 ml attained the highest growth parameters of rice plants. Inoculation with Azospirillum increased the uptake of N, P and Zn. Nitrogenase activity and total count of Azospirillum were also estimated. Zine-enriched azolla gave higher values of plant height, number of productive tillers, grains and straw yields than zinc sulphate. Zinc-enriched azolla (0.04 g/ pot) gave higher values of N, P and Zn uptake by rice plant than the other tested treatments.

 

5/28 CHARACTERIZATION OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS STRAINS ISOLATED FROM FOOD AND CLINICAL SAMPLES IN BENI-SUEF CITY, EGYPT.

Shaaban A. Hashem, Magdy A. Amin*, Aml E. Saafan** and Ahmed, Farag**.

Department of Microbiology and Immunology, Faculty of Pharmacy, Assiut University, Cairo University*, Beni-Suef University**, Egypt.

Between June 2007 to May 2008, a total of 342 Staphylococcus aureus isolate were recovered from 534 different food and clinical samples, collected from Beni-Suef city, Egypt. By the standard methods for detection of S. aureus, 201 (58.8%) were typical S.aureus and 141 (41.2 %) were found to bebiochemically atypicalS.aureus. Typical S.aureus isolates were nearly equal from milk and milk- products (7.8%) and from fast foods (7.0%). For clinical samples, the highest incidence for typical S. aureus was noticed from pus samples (16.0%), while the least incidence was in urine samples (4.3%). Three hundred and forty two S. aureus were examined for methicillin resistance by disk diffusion method and also subcultured on oxacillin resistance screening agar base )ORSAB( medium. The results of disk diffusion method revealed the presence of 109 (31.9%) methicillin-resistant S. aureus (MRSA) isolate but subculturing of the recovered isolates on ORSAB revealed that 98 (28.6%) resistant isolate can grow on it. Genotypic detection of MRSA isolates by detecting the presence of mecA gene using polymerase chain reaction (PCR) technique with phenotypic MRSA. It was found that there are 91 (83.5%) mecA gene carrying isolate and 18 (16.5%) isolate not carry this gene. Out of the 91 positive mecA gene, 14.2% of isolates were from food samples, 5.4% and 8.7% isolated from milk and milk products and from fast foods respectively. For clinical samples, 78 (85.7%) were MRSA, the highest frequency were from pus samples (27.4%) and at the age between 20 and 40 years (50%) and the least was among urine isolates (7.6%), and no isolate recovered above 60 years age.Antibiotic sensitivity testing was done with the most commonly used 13 antibiotics using disk diffusion method on the 91 MRSA. The highest percentage of resistance was to penicillin G (61.5%), then norfloxacin (36.2%), lomefloxacin (29.6%), Amoxicillin/clavulinic acid, clindamycin and azithromycin showed the lowest frequency of resistance (16.4%).There was no isolate resistant to vancomycin or amikacin (0%).

 

6/28 BACTERIOLOGICAL AND MOLECULAR STUDY ON MULTI-DRUG RESISTANT MYCOBACTERIUM TUBERCULOSIS IN EGYPT

Dalia Adel Salem, Zeinab Abd El-Khalek Ibraheim* and Mohamed Abbas Shemis**

Departments of Medical Microbiology, Biochemistry**, Theodor Bilharz Research Institute and Department of Medical Microbiology & Immunology, Faculty of Medicine, Cairo University*, Cairo, Egypt.

Rifampin (RIF) resistance in Mycobacterium tuberculosis (M. tuberculosis) is mostely associated with mutations in rpoB gene, which encodes the RNA polymerase–subunit. RIF resistance serves as a surrogate marker for the detection of multidrug-resistant M. tuberculosis (MDR-TB), as more than 90% of RIF resistant M. tuberculosis isolates are also isoniazid resistant. The INNO-LiPA Rif.TB assay for simultaneous identification of M. tuberculosis complex strains and the detection of RIF resistance has been evaluated with 15 MDR-TB isolates obtained from Egyptian tuberculous patients. The sensitivity and specificity for the detection of RIF resistance were and 93.3% and 100.0%, respectively when compared to conventional drug susceptibility methods. The codons most frequently involved in ropB gene mutations among MDR-TB were codon 531 (66.6%) and codon 526 (13.3%). The INNO-LiPA Rif.TB assay has the potential to provide rapid information that is essential for the effective management of MDR-TB.

 

 

7/28 VIRAL CO-INFECTIONS IN CHRONIC HEPATITIS C VIRUS PATIENTS

Ashgan Mohammed Zayed, Zeinab Abd Elkhalek Ibraheim* and

Ehsan Hasan Abdel Bari**

Departements of Medical Microbiology, Pathology**, National Hepatology and Tropical Medicine Research Institute. Departement of Medical Microbiology & immunology, Faculty of Medicine, Cairo University*, Cairo, Egypt.

The present work reports the prevalence of HCV/HBV and HCV/HCMV coinfections from Egypt -the country with the highest rate of HCV infection worldwide- where coinfection with these viruses is not yet documented.A total of 60 chronic HCV patients (positives to anti-HCV and serum HCV RNA) were included together with 20 anti-HCV-negative individuals as control group. Anti-HCV, HBsAg, HBcAb, Anti-HCMV (IgM & IgG) were assessed in the sera of patients and control group using commercially available ELISA kits. HCV RNA was detected using a commercially available semiautomated, PCR-based, qualitative assay. Liver biopsies were examined for histological lesions. HCV/HBV and HCV/HCMV coinfections were detected in 8.7% and 80% of HCV infected patients respectively. HCV positive patients investigated herein do not have an increased risk of exposure to HBV or HCMV infection as compared to HCV-negative individuals.Highly significant values of necroinflammation, fibrosis, and steatosis scores were detected in HCV/HBV and HCV/HCMV coinfected patients compared to HCV monoinfected patients. The risk factors of HCV/HBV and HCV/HCMV coinfection were found to be similar to those of HBV, HCMV or HCV monoinfection. HCV/HBV and HCV/HCMV coinfections are of clinical relevance, they increased the severity of histological lesions. More in-vitro studies are required to understand the viral interference in dually infected patients.

 

 

8/28 EVALUATION OF TEA TREE OIL AS AN ADJUNCTIVE TREATMENT OF CHRONIC PERIODONTITIS: A CLINICAL, BACTERIOLOGICAL AND IMMUNOLOGICAL STUDY

Eman A. Labah, Amal A. Al-Hessy*, Fatma A. El-Nomany* and Nehal M. Kadoom*

Microbiology & Immunology Department, Faculty of Medicine and Oral Medicine, Periodontology, Oral Diagnosis & Oral Radiology Department, Faculty of Dentistry*,Tanta University, Egypt.

Periodontal disease is recognized as a major public health problem throughout the world and is the most common causeof tooth loss in adults. This study evaluated tea tree oil containing gel releasing system in treatment of periodontitis in comparison with the mechanical and doxycycline hyclate gel therapies being utilized. The study detected the changes of clinical parameters (plaque index, plaque pocket depth and bleeding on probing), bacterial load indicated by BANA test and the level of interleukin-I b , during the course of treatments in all studied groups. The results of the present work showed that Tea tree oil containing gel delivery system had long term potent anti-inflammatory and antibacterial effects that improve the clinical parameters in patients with periodontitis more than so do doxycycline hyclate gel. It would be a unique, non toxic agents that would be useful addition to the current range of chemotherapeutic periodontal treatment options.

 

9/28 PREVALENCE AND ANTIBIOTIC RESISTANCE PATTERN OF AMPC AND EXTENDED SPECTRUM BETA LACTAMASES AMONG NOSOCOMIAL E. COLI IN TANTA UNIVERSITY HOSPITAL

Eman A. Labah and Ibtesam K. Afifi

Microbiology & Immunology Department, Faculty of Medicine,

Tanta University, Egypt

Antibiotic resistant mechanisms are the most worrisome problem in treating bacterial infections.Unreasonable use of the third generation has led to worldwide emergence of ESBL and AmpC beta lactamases that are responsible for treatment failure and outbreaks of nosocomial infections. This study aimed to detect the frequency of both resistant mechanisms in nosocomial E. coli and to test the efficiency of new discs for their detection. Isolated strains were tested for antibiotic sensitivity and those resistant to one or more of third generation cephalosporins were tested for ESBL by modified disc synergy test (MDDS) and for AmpC by modified three dimensional test. Further testing of the strains by Amp C and ESBL detection discs was done. Out of the 88 total E.coli isolates tested, 62 (70.5%) were resistant to one or more of third generation cephalosporines, 53 (60.2%) out of them were ESBL positive by MDDS test and 13(14.77%) were AmpC positive by modified three dimensional test. By AmpC and ESBL detection discs, 55 out of the strains were ESBL positive, 12 were AmpC positive and both enzymes was detected in 6 cases (6.8% of total). Antibiotic resistance pattern of most strains showed multidrug resistance to three or more antimicrobial classes while all of them were sensitive to imipenem. In conclusion, E.coli isolated from our hospital is mostly mutidrug resistant, produce ESBL in large percentage and AmpC is another emerging mechanism of resistance with coexistence of both mechanisms in some cases. ESBL and AmpC discs provide a cost-effective alternative approach for clinical microbiology laboratories for routine susceptibility testing with simultaneous detection of both resistance mechanisms. Thus continued monitoring of drug resistance during susceptibility testing of each organism and measures to restrict the indiscriminative use of cephalosporins through proper antibiotic policy is largely recommended to minimize the emergence of this multiple beta-lactamase producing organisms.

 

 

10/28 TRANSFORMING GROWTH FACTOR-BETA-1 (TGF-ß1) CODON 25 POLYMORPHISM IN HEPATITIS C VIRAL INFECTION

Wafaa K. Mahdi, Mona A. Esmail, Mohamed S. Mohamad, Hala I. Mohamad* and Soheir A. Latif**

Microbiology and Immunology Department, Faculty of Medicine, El-Minia University.

*Tropical Medicine Department, Faculty of Medicine, El-Minia University

**Clinical Pathology Department, National Cancer Institute, Cairo University

 

Hepatitis C virus (HCV) is a major cause of chronic liver disease. Cytokines secreted in response to cell injury have a central role in the pathogenesis of liver fibrosis. TGF b1 is a prosclerotic cytokine implicated in fibrotic processes. There are two genetic polymorphisms in the DNA sequence encoding the leader sequence of the TGFb1 protein, located at codon 10 (either leucine or proline) and at codon 25 (either arginine or proline).The TGF-ß1 genotype at codon 25 was determined in 60 patients {40 patients with chronic hepatitis, 20 patients with hepatocellular carcinoma (HCC)} and in 20 healthy control subjects. DNA extraction of TGF-ß1 was performed and amplified by real time PCR. There is a significant difference of gene polymorphism both in chronic hepatitis C patients and HCC with heterogonous alleles in 57.5% of chronic hepatitis patients and 65% of HCC patients. Significant differences were detected between the TGF-ß 1 polymorphism at codon 25 and the stage of hepatic fibrosis. Our findings suggest that TGF-ß1 codon 25 polymorphism could be a genetic factor associated with HCV infection and our results indicate that the heterozygous ArgPro genotype of codon 25 predicts significantly with increase degree of liver fibrosis of chronic hepatitis C than the homozygous ArgArg genotype.

 

 

11/28 A PILOT STUDY ON SOME DISINFECTANTS CONTROLLING WATER BORNE BACTERIAL CONTAMINATION AND BIOFILM FORMATION IN DENTAL UNIT WATER LINES

Ibtesam K. Afifi, Nagwa A. Ghoname*, Amina M.El-Hosary* and

Mohammed H. El Bayoumy**

Departments of Microbiology & Immunolgy, Faculty of Medicine, Tanta University.

Pedodontics* and Oral Health & Preventive Dentistry**, Faculty of Dentistry,                  Tanta University

Bacterial contamination and biofilm formation are the most important infection control challenges in dental unit waterlines (DUWLs). Detachment of microorganisms from dental unit biofilm flushed into the oral cavity could theoretically infect the patient. Splashes and aerosols from dental procedures may possibly infect health care personnel as well. The aim of this study was to assess the level of DUWLs bacterial contamination in Pedodontic clinic (assessment stage) and to compare between the efficacies of three disinfectants used to control it (pilot stage). Water samples were obtained from tap water, high speed handpieces (HSHP) and air water syringes (AWS) waterlines of Pedodontic clinic and cultured for evaluation of bacterial contamination. Then the same dental units were divided into four groups (untreated control, Calbenuim, IGN-Calbenium and A-dec ICX groups), and after application of disinfectant regimens, water samples were collected aseptically at mid working time daily, cultured and bacterial colony forming units (CFUs) were counted. Specimens from an evaluated dental unit and from each one of the four groups were examined by Scanning Electron Microscope (SEM). Water samples from HSHP and AWS waterlines of the dental units in the assessment stage showed high level of contamination (915665.63 and 627664.38 CFU/ml respectively). In the pilot stage, in contrast to control group, there was significant decrease in bacterial counts in disinfectants groups from the first day onwards. These bacterial findings were confirmed by SEM examination which revealed biofilm in both assessment and control groups with no evidence of its formation in the disinfectants groups. Conclusions: The water system of the dental units in Pedodontic clinic is highly contaminated when compared to American Dental Association goal (ADA), so, routine disinfection of this water is highly recommended. The tested disinfectants are effective in improving the quality of dental unit water, but further wide scale investigation should be conducted to evaluate the best, cost-effective disinfectant that maintains its effect on extended use.

 

 

12/28 PROGRESSIVE MACULAR HYPOMELANOSIS PATHOGENESIS AND TREATMENT: A RANDOMIZED CLINICAL TRIAL

Azza M. Hassan, Khaled A. Moustafa*, Fatma A. Abd Rabbou**

and Mohamed M. Gamei**

Departments of Microbiology & Immunology, Histology*, Dermatology & Venereology**, Faculty of Medicine, Tanta University

Progressive macular hypomelanosis (PMH) is a cosmetically disturbing skin disorder that is poorly understood, often misdiagnosed with unknown prevalence and indefinite treatment. The aim of the present study was to clinically outline PMH and study its pathogenesis. Based upon the literature suggesting improvement of PMH with antibiotic therapy, different treatment modalities were tried to reach the best treatment option. This study carried out from June 2009 to March 2010 and included 12 newly diagnosed PMH selected patients who attended Tanta University Hospital outpatient clinic of Dermatology and Venerology. Patient's lesions were subjected to wood's lamp and potassium hydroxide (KOH) examination as well as biopsies used in histological, immunohistochemical, bacteriological and electron microscopic examination. Additionally, a randomized clinical trial with different treatment modalities was applied with a follow up period of 6 months. The patient's mean age was 25±10.8 with female predominance (66.7%). Although this study showed the presence of mature melanocytes with no apparent difference in the number of S-100 protein-positive cells between the hypopigmented and normal skin, reduction of epidermal melanosomes, melanin granules concentration and tyrosine activity together with difference in distribution of S100 proteins in hypopigmented than normal skin was demonstrated. Additionally, abnormal distribution of tonofilaments was also noticed inside keratinocytes with presence of disintegrated melanosomes and increased apoptosis. Of notice, P. acnes were detected in hair follicles of 83.3% hypopigmented lesions. Regarding treatment strategy, administration of local and systemic antimicrobial treatment with narrow band ultraviolet B (NBUVB) phototherapy for 3 months was the best treatment modality without any manifestation of recurrence within 6 months follow up period. The present study suggested that the etiology of PMH is multifactorial where genetic predispotion, the presence of P. acnes and hormonal imbalance play the main role. Additionally, administration of local and systemic antimicrobial treatment with NBUVB phototherapy for 3 months is an effective treatment regimen for PMH.

 

 

13/28 SERUM PROCALCITONIN ESTIMATION AS A DIAGNOSTIC BIOMARKER FOR NEONATAL SEPSIS

Mohammad Saleh, Mona F. Salama*, Enas Hammad* and Basma Shouman**

Department of Medical Microbiology and Immunology, Damietta Faculty of Medicine, Al-Azhar University, Departments of Medical Microbiology and Immunology* and Mansoura Children Hospital**, Faculty of Medicine,

Mansoura University.

Neonatologists have increasingly faced troubles of accurately identifying neonatal sepsis. The clinical signs are unclear, non specific and there is no single reliable marker of infection existing. Procalcitonin (PCT) is a promising marker because of its sensitivity and in particular its specificity. It differentiates between infection and inflammation while the acute phase proteins such as C-reactive protein (CRP) cannot do. The chief objective of this study was to appraise the role of PCT as a diagnostic marker for neonatal sepsis comparing its results with CRP.In this study, 94 neonates with clinically suspected sepsis admitted to neonatal intensive care unit (NICU) in Mansoura University Children Hospital were included. Based on blood culture positive results, they were divided into 2 groups: proven sepsis and clinical sepsis. Comparing the 2 groups, a threshold value, sensitivity and specificity for PCT and CRP were established using receiver operating characteristic (ROC) curves. Of 94 neonates with clinically suspected sepsis, 36 (38.3%) were proven sepsis group and 58 (61.7%) were clinical sepsis group. Cutoff values with optimum diagnostic efficiency were 2.5 ng/ml for PCT (sensitivity 97.2%, specificity 98.3%) and 3.5 mg/dl. for CRP (sensitivity 97.2%, specificity 86.2%). In conclusion this study confirms significance of PCT assay as valuable tool for diagnosis of neonatal sepsis.

 

 

14/28 ADENOTONSILLAR TISSUES AS A RESERVOIR OF HELICOBACTER PYLORI INFECTION IN EGYPTIAN CHILDREN

Soheir S. Maklad, Enas K. Abo Elmagd, Hala Alsafi*, Fatma M. Abdel Gaber** and Tawfik A. Al-Kholy**

Microbiology, *Pathology and **ENT Departments, Faculty of Medicine for Girls, Al-Azhar University.

This study was planned to determine (i) possible Helicobacter pylori (H. pylori)colonization on tonsils and adenoids of children underwent adenotonsillectomy (ii) seroprevalence of H. pylori infection in those children. Biopsy specimens from 36 children with chronic tonsillitis and/or adenoiditis undergoing adenotonsillectomy were examined; 19 tonsil (group I) and 17 adenoid specimens (group II). The presence of H. pylori antigen in tissues was detected by immunohistochemical (IHC) staining and correlated with serological detection of H. pylori specific IgG by Enzyme Linked Immunosorbent Assay technique (ELISA). Ten normal control serum samples were also included (group III). Histological and IHC study showed that (52.63 %) and (52.94 %) out of the 19 tonsillar and 17 adenoid specimens respectively had manifestations of both infection and follicular hyperplasia or hypertrophy and were also positive for H. pylori antigen with intensity degree ranging from + to +++. There was insignificant difference (P>0.05) in H. pylori antigen positivity between tonsillar and adenoidal groups. Specific IgG to H. pylori was detected in 89.47 %, 64.71% and 30 % of group I, group II and normal controls respectively. The mean concentrations of specific IgG (ng/ml) in patient groups and controls were (76.72±36.88), (66.81±46.04) and (28.26±21.90) respectively. A highly significant differences (P<0.001) were observed in the frequency of patients with positive IgG and in the mean concentrations of specific H. pylori IgG between patient groups and controls. Similarity was found between IHC and serologic results, since (52.63 %) and (10.53%) of tonsils were positive and negative respectively by both techniques. Additionally, (41.18%) and (23.53%) of adenoid tissue gave positive and negative results respectively by both techniques. Conclusions: The results of this study support the notion that tonsils and adenoids may be a possible reservoir for H. pylori in children.

 

15/28 A STUDY ON PRESERVATIVES USED IN RECOMBINANT HUMAN INSULIN PREPARATIONS

Mostafa A. Baraka and Mohamed A. Ramadan

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

The multidose sterile recombinant human insulin market products require the addition of effective compatible preservation system to re-establish sterility after each use. However, the bacterial populations in the environment are frequently exposed to stresses, so those cell. Develop survival strategies to adapt with these conditions. In this study, the preservatives systems in-use with recombinant human insulin market products, are tested for their antimicrobial effectiveness towards mixed environmental strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa isolated from the manufacturing area using classical challenge test and also the antimicrobial effectiveness towards the induction and the viability of viable but non culturable (VBNC) cells of Enterococcus faecalis bacteria by measuring electron transport system (ETS) activity.

 

 

16/28 DEVICE – RELATED, BIOFILM PRODUCING STAPHYLOCOCCUS EPIDERMIDIS IN NEONATAL INTENSIVE CARE UNIT

Mohammad Saleh and Lotfi Abd-Elfatah*

Departments of Medical Microbiology and Immunology and Pediatrics*, Damietta Faculty of Medicine, Al-Azhar University

Biofilm producing Staphylococcus epidermidis (S. epidermidis) can cause severe infections in neonates with indwelling medical devices. Biofilm formation is considered a major virulence factor for S. epidermidis and renders the isolates highly resistant to antibiotic treatment. We evaluated the role of S. epidermidis in device associated infection in neonatal intensive care unit (NICU). Biofilm detection was achieved by polymerase chain reaction (PCR) which used for detection of icaA and icaD genes as a gold standard method comparing it with 3 phenotypic assays; tissue culture plate (TCP), Congo red agar (CRA) and haemagglutination (HA) assays. In this study, S. epidermidis was responsible for 31.3% of catheter related infections in NICU; it was isolated from 37.12%, 18.5% and 13% of peripheral venous catheters (PVC), central venous catheters (CVC) and urinary catheters respectively. IcaA and D genes were detected in 71.9% of isolates. Phenotypic detection of biofilm formation revealed that HA assay was the most sensitive (100%) method followed by CRA (85.3%) and lastly TCP assay (68.3%). Prolonged hospital stay and Prolonged catheterization were markedly associated with biofilm producing S. epidermidis device associated infection. More resistance to antibiotic was noticed in biofilm producing S. epidermidis isolates than non-producing isolates. In conclusion, S. epidermidis was responsible for one third of catheter related infections in NICU. The risk of infection with biofilm producing S. epidermidis increased with prolonged hospitalization and catheterization rendering strains more resistant to antibiotics. HA assay might be a reliable method for detection of biofilm formation as compared to CRA and TCP assays.

 

 


17/28 PREVLENCE OF EXTRAHEPATIC MANIFESTATIONS IN CHRONIC HEPATITIS C: AN EGYPTIAN CROSS-SECTIONAL STUDY

Eman I. Elhawary, Gamal F. Mahmoud, Mai El-Daly*,Fatma A. Mekky**,Gamal Esmat*** andMohamed Abdel-hamid*

Microbiology Department, Faculty of Pharmacy, Minia University; *Viral Hepatitis Research Laboratory; ** Community, Environmental and Occupational Medicine, Department, Faculty of Medicine, Ain Shams University and *** Tropical Diseases, Faculty of Medicine, Cairo University.

The aims of the present study were to assess the prevalence of clinical and biologic extrahepatic manifestations (EH) and to identify associations between these manifestations among Egyptian patients. Several extrahepatic manifestations have been reported in the natural history of hepatitis C virus infection (HCV). The prevalence of clinically significant EH manifestations is relatively low, but can be associated with significant morbidity and even mortality. An awareness and recognition of these manifestations is of paramount importance in facilitating early diagnosis and management of these complications. A cross sectional study was conducted. Two hundred and eighty-nine patients having chronic HCV infection were interviewed in this research. Laboratory investigations for liver function, cryoglobulin profile, blood sugar, rheumatoid factor, antinuclear antibody (ANA) and HCV-PCR were performed. Association between cryoglobulin-positivity, diabetes positivity and other manifestations were identified using χ2 and Fisher’s exact test. Risk factors for the future development of extrahepatic manifestations were assessed by logistic regression analysis. The prevalence of dermatologic, rheumatologic, diabetes, autoantibodies, and cryoglobulins were assessed. Demographic, biochemical, immunologic, virologic, and liver histologic factors associated with the presence of EH manifestations as well as the prelevance of cryoglobulinemia and diabetes to the reported manifestations were identified. One hundred eighty three (63.3%) HCV cases showed at least one EH manifestation. Cryoglobulinemia was found positive in 22 patients (7.6%). Rheumatological manifestations were diagnosed in 18.3% of the study group. The major symptom was xerostomia (9.3%) and chronic fatigue syndrome (8.7%). Regarding the dermatological consultation, pruritis (3.8%) followed by lichen planus (2.1%) and psoriasis were the most common dermatological diseases diagnosed by dermatologic consultants in the studied cases. Diabetes was diagnosed in 40 (13.8%) patients. In Chronic HCV genotype 4 infection, the prevalence of extrahepatic manifestations was 63.3%. The most frequent manifestations were rheumatoid factor (31.5%), rheumatological manifestations (18.3%), diabetes (13.8%), dermatological manifestations (9.7%), cryoglobulinemia (7.6%) and antinuclear antibodies (4.2%).There was a significant difference between EH cases and normal HCV cases regarding age, gender, clinical and sonar findings, education level and bilirubin levels in univariate analysis. Advanced age and residence location were risk factors for developing diabetes; gender and elevated Bilirubin levels were predictors of rheumatological EH manifestations; arthralgia and level of education were predictors of dermatological EH manifestations. It is therefore recommended that patients with any of these conditions be tested for hepatitis C and, conversely, that signs and symptoms of these conditions be sought in patients with known chronic hepatitis C infection.

 

 


18/28 DETECTION OF HUMAN PAPILLOMAVIRUS (HPV) TYPE 16 AND IMMUNOHISTOCHEMICAL EXPRESSION OF FAS AND FASL AND THEIR CORRELATION WITH CLINICOPATHOLOGICAL PARAMETERS IN BREAST CARCINOMA  

Mona A. Khattab, Nermeen S. Youssef*, Abeer A. El-Sayed, Alaa A. Aliand Taghreed H. El-Khashaab

Departmentsof Medical Microbiology & Immunologyand Pathology*, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

Background and objective: Breast cancer is the most frequently diagnosed cancer worldwide. Many risk factors have been associated with the pathogenesis of it including family history, hormones, and cigarette smoking. Some types of viruses especially human papillomavirus (HPV), might be involved in its pathogenesis.Tumor infiltrating lymphocytes (TILs) are capable of infiltrating tumors but, there are several mechanisms obscuring their cytotoxic function e.g. apoptosis of TILs, which is mediated by Fas Ligand (FasL; CD95L)/ Fas(CD 95). The current study was designed to detect HPV16 E6 gene, assess Fas and FasL expressionand evaluate their correlation with clinicopathological parameters and prognostic value in breast tumors. Materials and Methods: This study was conducted on archived cases of breast lesions: 30 breast carcinomas and 15 benign breast lesions. Formalin fixed paraffin embedded sections from all cases were examined by PCR to detect the presence of HPV16 E6 gene and immunostained for Fas and FasL using the Avidin-Biotin immunoperoxidase complex technique. Results: HPV-16 DNA was detected in 30% of breast carcinomas and in none of benign breast lesions. Statistically, there was a significant relationship between detection of HPV-16 and breast carcinoma with advanced stage (p=0.012), positive L.N metastases (p=0.021), negative estrogen receptor (ER), progesterone receptors (PR) status (p<0.001) and positive HER/2neu status (p=0.012). Fas expression was detected in 93% of benign breast lesions and 60 % of breast carcinomas with a statistical significant difference (p=0.048). However, FasL expression was detected in 13% of benign breast lesions and 56.6% of breast carcinomas with a statistical significant difference (p=0.014). A statistical significant relationship was detected between Fas expression and breast carcinomas with small size (p=0.006), low grade (p=0.028), early stage (p<0.001), negative lymph node metastases (p=0.025) and positive (ER) status (p=0.048). There was a statistical significant relationship between FasL expression and breast carcinomas with high grade (p=0.049), advanced stage (p< 0.001), positive lymph node metastases (p < 0.001) and negative ER status (p=0.0257). There was a significant inverse linear correlation between Fas and FasL expression in breast carcinoma (p<0.001). There was a statistical significant inverse relationship between detection of HPV-16 and Fas expression (p=0.018). Conclusion:HPV detection was correlated with poor prognostic factors in breast carcinoma. Down-regulation of Fas expression together with up-regulation of FasL could be associated with   poor prognosis of breast cancer.

 

 


19/28 ERADICATION OF ENTEROCOCCUS FAECALIS AND CANDIDA ALBICANS REFRACTORY TO ENDODONTIC TREATMENT BY MECHANICAL INSTRUMENTATION AND IRRIGATION WITH ANTIMICROBIAL SOLUTIONS

Hamido M. Hefni, Abd El-Naser A. Almoghazy, Mohamed S. Ali, Mervat I. Fawzy*and Maha A. El-Hosiny*

Department of Microbiology and Immunology, Faculty of Pharmacy (Boys),

Al-Azhar University, Cairo, Egypt.

*Department of Endodontics, Faculty of Dental Medicine (Girls), Al-Azhar University, Cairo, Egypt

Using hand and rotary instrumentation techniques, two different irrigation solutions, namely 2.5% Naocl + EDTA. with and without minocycline, citric acid and polysorbate 80 (MTD) were investigated for their capabilities to significantly decrease the CFUs of two of the most resistant microorganisms ,namely Enterococcus fecalis and Candida albicans. One hundred and thirty recently extracted human upper central incisors were used in this study. After crown sectioning and standardizing root canals to 15 mm followed by autoclaving ,root canals were seeded with standard inoclum, one of the two tested microorganisms. Six roots were assigned for each irrigation technique for a specific microorganism with 10 roots were used as control sterile and 10 roots used as control septic i.e. irrigated with 0.9 Nacl saline only. The number of CFUs of each microorganism at zero time and after each irrigation step as well as   at the end of the experiment were statistically analyzed. Statistical analysis showed the highest persistent significant reduction in microbial CFU were in groups tested with final MTD treatmen., Neither significant difference was observed due to instrumentation types technique nor due to 2.5% Naocl +EDTA as a final irrigation step.

 

 

20/28 POTENTIALITY OF SOME ALKALIPHILIC FUNGI ISOLATED FROM EGYPTIAN SOIL AS PRODUCERS OF ALKALINE DETERGENT ENZYMES

Mohamed Ghareib, Amal A. Khalil, Hossam S. Hamdy and Medhat Abu Tahon

Department of Biological and Geological Sciences, Faculty of Education,

Ain Shams University, Roxy, Cairo, Egypt.

Thirty four species of fungi were examined for their ability to grow under alkaline conditions. Nine species to the genera Alternaria, Aspergillus, Fusarium, Peacilomyces and Penicillium exhibited growth profile of facultative alkaliphilic fungi. These fungi had the potentiality to produce different alkaline detergent enzymes. Aspergillus flavus, Fusarium oxysaporum, Paecilomyes variotii and Penicillium chrysogenum were recorded as the best producers for the alkaline a-amylase, cellulase, lipase and protease, respectively. Growth and production of alkaline detergent enzymes by these four fungi were favorably affected by addition of sodium chloride to the fermentation media. The selected four fungi were categorized as alkaliphilic-halotolerant organisms. Washing properties of the enzyme mixtures of the four fungi were also investigated. The detergent enzyme complex of P. chrysogenum retained its stability in presence of the commercial detergents and gave the best results of washing test.

 

 


21/28 RELATIONSHIP BETWEEN HUMAN PRAY AND MICROORGANISMS

Nemmat A. Hussein1,3 and Abeer A. Abu-Zaid 2,3

1Botany Dept., Fac. of Science, Assiut University, Assiut, Egypt,

2Food Technology Rese Inst., Agric Research Cent., Cairo, Egypt

3Biology Dept., Fac. of Science, Taif University, Saudi Arabia

The biodiversity of fungi and bacteria was studied in samples taken from hand, oral cavity, nasal cavity, ear and foot of 10 non prayers and others prayer people, before and after their Wudu. Fungi were isolated on potato dextrose agar (PDA) and sabouraud dextrose agar (SDA), while bacteria were detected on nutrient agar (NA) medium. In general, it was detected that bacterial organisms were dominant in tested samples than fungal organisms. The highest bacterial counts were registered at swabs from foot and hands (respectively) of non prayers. While the lowest bacterial counts were detected at samples from oral and nasal cavities of prayers after wudu. On the other hand, the higher fungal contaminations were detected at swabs from foot (on SDA) followed by oral cavity (on SDA and PDA), while the lower fungal contaminations were registered at foot swabs from prayers after wudu. The most common fungal species were Candida albicans, other yeasts, Aspergillus flavus, A. niger and Fusarium poae in most tested samples.

 

 

22/28 BK VIRUS AND CD 4+ CD 25+ T REGULATORY CELLS IN CANCER PATIENTS.

Mona O. Abbas, Manal M. Yassin*, Abeer A. El-Sayed*, Walaa A.Sadek* and Hany M. Abd El Azziz**

Microbiology & Immunology Department, Faculty of Medicine, King Abd El Aziz University, Gedda, Kingdom Saudia Arabia.

Departmentsof Medical Microbiology & Immunology* and Radiation Oncology & Nuclear Medicine**, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

Background: K virus (BKV) is a member of the polyomavirus family. After primary infection, BKV establishes a lifelong latent infection in many organs. An association of BKV DNA with a variety of human tumors was reported.   CD4+CD25+ T regulatory (Treg) cells maintain immunologic self-toleranceand suppress immune responses to tumors, transplants, andinfectious agents. Aim: the aim of this work was to detect BK virus and measuring CD4+ CD25+ T regulatory cells in cancer patients and study the relation between them and cancer. Patients and methods: This study was conducted on 45 patients (subdivided into three groups (n=15 for each): urinary tract, breast and hematological cancer patients) and twenty controls. They were admitted to Ain Shams University Hospital from May 2007 to March 2008. BK virus DNA was detected by conventional polymerase chain reaction (PCR) and real time PCR. CD4+CD25+ Treg cells percentage was detected by flowcytometry. Results: BK virus was detected by conventional PCR and real time PCR in six and eleven patients (respectively), in urinary tract cancer group only, which revealed statistically significant and highly significant difference (P value = 0.003, P value < 0.001 respectively) between the three cancer groups. Also, the difference between urinary tract cancer patients and controls for detection of BK virus   by conventional PCR and real time PCR was statistically significant and highly significant difference (P value = 0.003, P value<0.001 respectively). Sensitivity and specificity of Real time PCR in detection of BKV DNAwere higher statistically significant when compared to conventional PCR (P < 0.001). Patients of the three groups of cancer showed highly statistical significant elevation in their mean percentage of CD4+CD25+Treg cells compared to healthy controls(p<0.001). Conclusion: Association of BKV DNA with urinary tract cancer suggests an important oncogenic potential in the pathogenesis of cancer. The significant elevated CD4+CD25 +Treg cell percentage in cancer patients might enable the cancer cells to evade the host immune response. So targeting Treg cells provides a novel targeted immune interventions and prevention to treat and prevent cancer.

 

 

23/28 EVALUATION OF A CYTOMEGALOVIRUS (CMV) IGG AVIDITY TESTING TO DISCRIMINATE BETWEEN ACTIVE AND LATENT CMV INFECTION

Mohamed M. El Sawy, Reem A. Harfoush*, Nesrine F. Hanafi* and

Sara M. Abdel Hameed**

Department of Clinical Pathology, Faculty of Medicine, Alexandria University, Egypt

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

**Department of Medical Microbiology and Immunology, Alexandria University, Students' Hospital, Egypt.

Differentiation between active & latent CMV is not easy to be achieved inspite of being of high concern to the clinicians especially in critical cases such as pregnant females and transplanted patients. It is proposed that testing IgG avidity could be a reliable cost effective method to solve this problem. The aim of this study was to evaluate the performance of a fully automated chemiluminescent immunoassay for determination of anti-CMV immunoglobulin G (IgG) avidity in comparison with real-time PCR for detection of CMV DNA in blood. Our study comprised samples from 80 patients (23 women suffering habitual abortions, 43 pregnant female performing routine investigation in the first trimester and 14 kidney transplant recipient patients). The IgG positive sera were tested for IgG avidity using Liaison CMV IgG avidity assay, while real time PCR was performed on the whole blood. It was found that low avidity indices (AIs <20%) were detected in 17.5% of patients, intermediate AIs (21%-30%) in 7.5% of patients, and 75% had high AIs (>30%). So, the ability of the avidity assay to exclude a recent CMV infection was 82.5% (AI > 20%). Results of PCRcorrelated well with those of avidity, since all patients that had low AIs were PCR positive, and PCR was negative in the remaining patients with intermediate and high AIs. Consequently, the avidity assay showed 100% sensitivity and an overall specificity of 90% when compared with PCR. So, the combination of these two methods based on different approaches appears to be a simple system that can be used to improve serological diagnosis.

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