A-61 IN VITRO AND IN VIVIO EFFECTS OF SALVADORA PERSICA (MISWAK- SIWAK) ON HERPES SIMPLEX VIRUS INFECTION
MAHMOUD Y.M. TAHA, PH.D
Department of Dental Basic Sciences, College of Dentistry, Mosul University, Mosul –Iraq
Email: tahadent04@yahoo.com
Aim: The present study was conducted to investigate the effect of Salvadora Persica extract on HSV-1 infection both in vitro and in vivo in the mouse model system.
Materials and Methods: Ethanolic extract of Salvadora Persica was used at different concentrations. BHK cells were grown in Eagles medium were used for virus isolation and titration using PFU/ml. The effects of different concentrations of Salvadora Persica on viral growth in BHK cells as well as cytolytic activity of HSV-1 were evaluated at different time post infection. The therapeutic efficacy of Salvadora Persica in vivo was studied in mice. Lesions were scored and viral isolation from infected skin and ganglia was titrated on BHK cells.
Results: Salvadora Persica inhibited the replication of HSV-1 in BHK cells as well as the cytolytic activity of cell free virus. Topical application of Salvadora Persica on the skin of mice infected with HSV-1 reduced the development of cutaneous lesions and the viral titers in the skin and ganglia were also reduced.
Conclusion: The results of this work may be beneficial for the treatment of recurrent oral herpes infections.
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A-62 Pulmonary aspergillosis in immunocompromised patients
Hoda Abd el.Monem Naguib, M.D.
Department of Microbiology &Immunology Faculty of medicine, Mansoura university, Egypt
Email: Dr.Merit@gmail.com
Objectives: This work was designed to study the magnitude of invasive pulmonary asperigellous problem, determine the risk factors, the underlying conditions and proper methods for its diagnosis.
Materials and Methods: The work was included a group of 37 immunocompromised patients admitted to various departments in Mansoura University Hospitals (Chest, Hematology, Radiotherapy, Renal Dialysis and Urology &Nephrology Center).Samples were collected and managed according to the standard laboratory procedures; sputum samples,BAL,FNA,EDTA –anticoagulant blood and serum samples.
Results: Pulmonary aspergillosis was identified in (23.3%) of clinically suspected cases, patients with neutropenia more than 12 weeks had a risk for PA in (73.7 %) of cases, anticancer chemotherapy in (29.7 %). Corticosteroids treatment in 37% of cases, thrombocytopenia in 35.7% of cases. The commonest aspergillus species isolated was A.fumigatus. Aspergillus terreus. A. versicolor and A.Flavus were also isolated fromIPA.Aspergillus niger was also detected in colonization cases. Diagnostic tests included direct microscopic examination of sputum, BAL and FNA their sensetivities were (33.3%, 77.7 and 100% respectively), sensitivity for culture of theses samples was (50%, 100% and 100% respectively) specificity was found (91%, 83.3% and 100% respictevely) Blood PCR has shown excellent sensitivity and specificity (100% and 94.7%) has shown 15% false positive result. BAL PCR has excellent sensitivity and moderate specificity (100% and 66.6%) also has false positive result is 38.8%)
Sensitivity and specificity of Elisa and PCR did not differ according to certainty level or clinical aspergillosis type.
Conclusions: Invasive pulmonary asperigillosis can be prevented in immunocompromised patients setting by proper identification, correction of risk factors and establishment of a policy for proper prophylactic anti-asperigillus antifungal. Although culture and direct KOH preparation has moderate sensitivity it has high specificity and it is cheap and easy. Although antigen detection is expensive, it is simple, easy; the result can be forwarded to the clinician within one day and has excellent sensitivity and specificity. DNA detection methods expensive, tedious difficult to be standardized, it can be adopted due to its excellent sensitivity and specificity. Pleural fluid, CT guided FNA and BAL are representative samples showing excellent sensitivity and specificity for KOH preparation, culture and PCR, yet not all patients especially of critical condition could obtain these invasive samples. Although each individual technique has its drawbacks, using them all results in an earlier and more definite diagnosis of IPA (a condition having non specific symptoms and should be included in the differential diagnosis of pneumonia, lung abscess, tuberculosis and bronchiectasis).
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A-63 Candida albicans; a novel approach to overcome its adherence property
Dr. Shama M.J. Saadaldin, Ph.D.
Senior lecture in Medical Microbiology College of Medicine Al-Mustansiriya University P.O. Box 14132 Baghdad, Iraq E mail: shama171063@yahoo.com
Objectives: The aim of this study is to introduce a procedure mimic the adherence of microorganisms to the plastic tube and to assess the suitable solution to irrigate these microorganisms.
Materials and Methods: Candida albicans broth was inoculated into 30 cm plastic tube for certain period of time to allow the adherence of microorganism to the inner surface of tube. Then the tubes were irrigated with distilled water, or tap water or different concentrations of sodium chloride (0.1-0.9%). The optic densities of the Candida albicans broth as well as the recovered irrigated solutions were spectrophotometrically monitored at 620 nm.
Results: The results showed that the percentage of Candida albicans which adhered to the inner surface of tube ranged between 0.27% and 30% of the initial inoculum. Tap water was superior than distilled water in removing the adherent Candida. Sodium chloride in concentrations 0.3% and 0.9% completely removed the attached Candida while the other concentrations showed inconsistent results. The ability of irrigated solution to detach the Candida albicans was not related to the initial inoculum concentration.
Conclusions: It concludes that irrigation with water or certain sodium chloride concentrations, which interfere with the fluidity of Candida albicans, is the simple and best method to eliminate the adherent microorganisms.
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A-64 Prevalence and antimicrobial resistance of ESBL producing bacteria in
Al-Assad University hospital- Lattakia
Mazen krawi –Nada kherbeck
Tishrin University- faculty of medicine- department of laboratory medicine- Lattakia-Syria
Email: mazenkrawi@hotmail.com
Objective: To servile the prevalence rate of ESBL strains from gram- bacilli (enterobacteriaceae and non fermenter) and to determine its antibiotic resistant.
Material and methods: A total of 350 isolates from different site tested: 189 E.coli, 105 Klebsiella, 21 Pseudomonas, 35 proteus. ESBL screening was done by double disc synergy test. Disk diffusion susceptibility test conducted to determent the sensitivity to antibiotic and statistic method (p value) to determine the significance of combination.
Results: 40.8% (143) of isolates were ESBL positive, of these 52%(74) were E.coli, 42%(60) klebsiela,4%(6) pseudomonas and 2%(3) were proteus. 57% of isolated klebsiella were ESBL positive, 39% of E.coli, 28% of pseudomonas and 8.5% of proteus. all isolated strains were susceptible to amikacine and imipenem, 26.5% of the ESBL positive isolates were resistant to gentamycin and 61% resistant to ciprofloxacin. Resistant to gentamycin and to ciprofloxacine was significantly higher in ESBL+ isolates (p≤0.001).
Conclusion: The results shoe high prevalence rate of ESBL+, klebsiella strains has the highest rat, after come E.coli, Pseudomonas and Proteus. a surveillance study for antibiotic use conducted in the same hospital shoes extensive use of cephalosporins third generation. ESBL+ strains have higher resistance to gentamycin and ciprofloxacin. All ESBL+ isolates were sensitive to amikacin and imipenem.
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A-65 Study of Microbiological Flora and Role of Primary Bacterial Cultures in Management of Open Fractures of Long Bones
Shiraz M Bhatty, Rajesh Paul, Harjit Kaur
Department of Orthopaedics, Christian Medical College and Hospital LUDHIANA – 141008. PUNJAB (INDIA) Email: shirazbhatty@gmail.com
Background: Microbiology of open fracture wounds is constantly changing. A clear understanding of the bacterial flora that could be expected is needed in order to administer a rational and effective antibiotic treatment for open fractures. The role and efficacy of primary bacterial cultures in management of open fractures is also debatable and needs further evaluation.
Methods: One hundred patients with one hundred and seven open fractures of long bones were studied prospectively, from March 1st 2001 till February 28th 2002. Wound swabs were obtained at pre-debridement, intra-operative, post debridement, 1st dressing/after 24hrs intervals, subsequently every week and sent for cultures. The infecting organism, its antibiotic susceptibility and its correlation between cultures at different stages was noted.
Results: An infection rate of 43.9% was noted. Most of the initial wound cultures, showed growth of Gram-negative organisms (76%), commonest being Pseudomonas (36%) and Acinitobacter(20.7%). However, majority of infections after 2nd week were caused by Gram-positive organisms. Staphylococcus aureus (93.5%) was the predominant Gram-positive organism.
None of the organisms grown on admission and pre-debridemet cultures eventually caused infection; however, 28% of cases with negative cultures eventually got infected. Post debridement cultures were positive in none. Among the cultures obtained at 1st dressing 40% of organisms grown eventually caused infection whereas 60% showed growth with different organism.
Conclusions: A shift in the bacterial flora occurs in compound fracture wounds from Gram-negative to Gram-positive organisms after the 2nd week. Cultures obtained at admission, predebridment, postdebridment and at 1st dressing or after 24hrs are not reliable indicators of subsequent wound infection.
Keywords: Open fractures, bacterial flora, wound cultures.
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A-66 Resistance Patterns and Risk Factors of Streptococcus pneumoniae Carriage among Healthy Jordanian Children
Rawaa Al-Kayali1, Hala Khyami-Horani1*, Adnan Al-Lahham2
German Jordanian University, School of Applied Medical Sciences, Biomedical Engineering, Amman, Jordan
Email: adnan.lahham@gju.edu.jo
Objectives: Isolation of Streptococcus pneumoniae from healthy children in Amman, Jordan.
Methods: Swabs from Nasal cavity were taken from 1002 healthy children. Isolates were analysed for antimicrobial susceptibility, serotyping, macrolide resistant genotypes and phenotypes.
Results: The overall carriage rate was 19.5% (n= 195 isolates). The percentage of resistance was as follows: Penicillin (91.8%), cefotaxime (29.2%), trimethoprim-sulfamethoxazole (66.7%), erythromycin (46.7%), clindamycin (19.5%), tetracycline (32.3%), chloramphenicol (6.2%) and ciprofloxacin (6.2%). 67 isolates (34.4%) were multi-resistant. (MIC50, MIC90) were as follows (μg/ml): Penicillin (1, 4), cefotaxime (1, 4), erythromycin (0.5, 32), and clindamycin (0.125, 32). PCR-based serotyping of the serotypes included in the 7v-PCV showed the following percentages: 19F (11.8%), 23F (9.7%), 9V (7.6%), 6B (6.7%), 14 (5.6%), 18C (4.6%) and 4 (2.6%). Among macrolide resistant isolates (n= 91), erm(B) was detected in 20.9% (n= 19), mef(A) in 53.8% (n= 49), both erm(B) and mef(A) 16.5% (n= 15) and 8.8% (n= 8) were negative for both determinants. Among the macrolide resistant isolates MLSB resistance and M- phenotypes were 41.8% and 58.2%, respectively.
Conclusions: The overall carriage of pneumococci in Jordan is low, but the resistance is high, nevertheless the spread of pneumococcal multi-drug resistance is worrisome.
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A-67 Antibiotic susceptibility in clinical isolates of Staphylococcus aureus strains isolated in Casablanca (Morocco),
Mohamed Elazhari1,2 * , Khalid Zerouali3 , Driss Elhabchi4 , Noureddine Dersi1 , Mohammed Hassar1,5 , Mohammed Timinouni5, Rachid Saile2 .
1: Laboratoire de Bactériologie Médicale, Centre de Biologie Médicale, Institut Pasteur du Maroc, 1 place Louis
Pasteur, Casablanca-20360, Maroc.
2: Département de Biologie, Faculté des Sciences Ben M'Sik,Université Hassan II, UFR Biologie et Santé,
Casablanca, Maroc.
Email: mohamed.elazhari@pasteur.ma
Objective: The aims of the present study was to make an update on the susceptibility of Staphylococcus aureus in Casablanca, and therefore to define the prevalence and the characterization methicillin-resistant strain.
Materials and methods: Clinical S. aureus isolates were collected from patients at the Institute Pasteur of Morocco and from 14 private laboratories located at Casablanca, from January 2007 to December 2008. Susceptibility testing was performed by the disk diffusion method on Mueller-Hinton agar plates. Strains expressed phenotypic resistance to cefoxitin were confirmed by polymerase chain reaction detection of the mecA gene, typing of the accessory gene regulator “agr” and the Staphylococcal Cassette Chromosome “SCCmec” has been made for mecA positive strains.
Results: In total, we have noted 31 different resistance pattern. On the other hand, among twelve strains (7,6%) witch have a low susceptibility to cefoxitin, five (3,2%) isolates were Borderline Resistant S.aureus “BORSA” or Modified Penicillin-Binding S. aureus “ MODSA”, and three (1,9% ) had the mecA gene with an agr type 1. Two of theme with type III SCCmec mercury negative and one with type I SCCmec.
Conclusions: The community strains of S. aureus circulating in Casablanca had a diversity of their resistance pattern. Only 1.9% of strains were resistant to methicillin by the presence of mecA gene.
Keywords: Casablanca; Community; Methicillin; Resistance; Staphylococcus aureus
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A-68 Prospective evaluation of the relevance of the antibiotic combinations in an internal medicine department
Thibaut Caruba, Brigitte Sabatier, Matta Matta*, Jean-Benoît Arlet, Brigitte Ranque, Loïc Capron, Jean-Luc Mainardi, Jacques Pouchot.
Lebanese American University, Byblos, Lebanon.
Email: drm_matta@yahoo.com
Objectives: The purpose of this study was to assess the relevance of antibiotic combinations according to available recommendations, and the impact of a clinical microbiology team.
Material and Methods: Prospective monocentric study of 12 month duration in an internal medicine department. All antibiotic combinations were assessed taking into account the infection type, the involved bacteria, the antibiotic nature, dosage and duration of treatment. A scientific committee classified each combination antibiotic therapy as: 1) “relevant to the existing recommendations”, 2) “adapted to the antibiogram in the absence of available recommendations”, 3) “irrelevant but with no adverse clinical outcome for the patient”, 4) “irrelevant with potential adverse clinical outcome for the patient”.
Results: Amongst the 87 antibiotic combinations prescribed, 67(77%) were relevant according to all available recommendations. Irrelevant combination therapy with and without potential adverse clinical outcome for the patients were 7% (6 cases) and 16% (14 cases), respectively. Reasons for non conformity included: prescription of a combination therapy while a monotherapy could have been sufficient or antibiotic therapy was unjustified (13 cases), prescription of antibiotics not adapted to the antibiogram (3 cases), prescription of antibiotics ineffective on the main bacteria encountered in the treatment of empirical infection (3 cases), and inadequately prolonged duration of antibiotic therapy (1 case). The advice of the mobile microbiology team has been done in 52% of the prescriptions and the appropriateness rate of prescription was then of 96% vs 57% (p<0.001) when the team was not solicited.
Conclusion: In this prospective study, 77% of antibiotic combinations were appropriate and relevant to the available recommendations. The advice of a microbiology team markedly improves this rate.
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A-69 Detection of mycobacterium tuberculosis in the saliva of patients having pulmonary tuberculosis
Gassan Yassen
E-mail: ygassan@hotmail.com
Tuberculosis is a serious disease caused by bacteria called mycobacterium tuberculosis. The disease is readily detected by demonstration of M.tuberculosis bacteria in clinical specimen.
The purpose of this study was to determine the efficacy of saliva as a sample for diagnosis the pulmonary tuberculosis by looking up for acid fast bacilli in direct smear and compare it with sputum and parotid saliva.
Pre-therapy group included 25 participant who diagnosed having pulmonary tuberculosis .Their ages range from (17-65) years.
Unstimulated saliva and parotid saliva was collected for direct smear for acid fast bacilli by Ziehl-Nelson acid fast stain .Five samples were inoculated on Lowenstein Jensen media and stonebrink media.
Concerning the M.tuberculosis, about 60% of unstimulated mixed saliva revealed positive acid fast bacilli, while all samples of parotid saliva showed negative acid fast bacilli. There was no significant relationship between the duration of sign and symptoms of disease and the detection of M.tuberculosis in collected specimens. The five samples of saliva which were inoculated on Lowenstein Jensen media and stonebrink media showed positive culture.
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A-70 Identification of Tn402 transposons, class 1 integrons and ISCR elements among endemic multi-drug resistant Klebsiella pneumoniae from Benghazi, Libya
El Salabi A, Toleman MA, Weeks J, Walsh TR.
Addresses: Department of Medical Microbiology, School of Medicine, Cardiff University, University Hospital of Wales, Cardiff CF14 4XN
E-mail: el_salabie@cardiff.ac.uk
Objectives: Klebsiella pneumoniae represents the most common Gram-negative pathogen in Libya in ICU wards and is becoming increasingly difficult to eradicate. As little is known of the antibiotic resistance in Libya, a program investigating antibiotic genes in K. pneumoniae was undertaken.
Materials and Methods: A total of 52 clinical and non-clinical (from hospital wards and operating theatres) were collected from Benghazi in 2008. Extended spectrum ��-lactamases (ESBLs) were detected using cefepime ESBL Etest. Colony blotting was first used to determine the presence of class 1 integrons, Tn402 transposons and ISCR elements. PCR experiments were performed to amplify integrons and transposons, and the amplicons sequenced and analysed. S1 and I-Ceu1 genomic digests of the whole DNA followed by probing was used to determine the genetic locations of resistance genes. PFGE was used to type the strains.
Results: 41/61 demonstrated resistance to cefotaxime, ceftazidime, aztreonam and also cefoxitin and cefepime. Of these 41 isolates, 26 were resistant to ciprofloxacin, 23 resistant to sulphamethoxazole /trimethoprim (SXT) and 35 resistant to gentamicin. Of the MDR strains 3/61 are fully resistant apart from carbapenems and 22/61 sensitive only to amikacin. 36/61 ESBL positive isolates possessed a class 1 integron correlating with the SXT resistance. 32/61 possessed Tn402 with the gene arrangement Int-dhfr-qac-TniC-orf8-TniB-TniA. The class 1 integrons contained aacA4. 5/61possesses ISCR elements and were not associated with any particular type of resistance. PFGE shows that the MDR isolates are the same strain type and were found in ICUs and local lakes. The Tn402 transposons were found on the chromosome and
on 6 plasmid sizes; 10, 15, 75, 200, 250 and 300kb.
Conclusions: These data shows an endemic K. pneumoniae MDR resistant strain in Benghazi ICUs and local lakes. All MDR strains carry a Tn402 transposon which is both plasmid and chromosomally encoded. As carbapenems are not always available in Libyan ICUs, these data shows a worrying problem.
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