VAAM-Jahrestagung 2012 18.–21. März in Tübingen

87measured by conventional enzymatic methods and micro immunefluorescence method. SPSS version 17 software was used for analyzing thedata.Results: Antibody test against Chlamydia pneumoniea was positive in 41patients (47.67%) and were diagnosed as seropositive. Confirming nosignificant difference between seropositive and seronegative patients inHDL (p=0.4), mean concentrations of Total cholesterol, LDL andTriglyceride were significantly higher in Chlamydia pneumoniaseropositive patients respectively. The mean concentrations of triglyceride,Total Cholesterol and LDL were P=0.003, P=0.001 and P=0.005. To seethe relationship among IHD, infection and lipid profile multivariateanalysis were done that, it was also following the univariate results.Conclusion: Thus, C. pneumoniae antibodies seem to correlate with analtered serum lipid profile considered to increase the risk ofatherosclerosis. This finding supports the proposal that C.pneumoniaeinfection may play a role in the pathogenesis of atherosclerosis (5). Insome cases, the infectious agents are found within the plaques and viableorganisms can be isolated suggesting a direct effect (4). Also, C.pneumoniae antibody positivity was independently associated withischemic stroke in elderly patients (6). Chronic Chlamydia pneumonieainfection was one of the risk factors for Ischemic heart disease, but toknow its exact role and mechanism, more studies are required.HMP004Role of Pseudomonas aeruginosa in nasocomial infections andapproach to its treatmentY. Hashemi Aghdam* 1 , S. Rahimi 2 , A. Moradi* 3 , F. Golchin* 41 Islamic Azad University, Medical Faculty, Young Researchers Club, TabrizBranch, Tabriz, Islamic Republic of Iran2 Islamic Azad University, Medical Faculty, Tabriz, Islamic Republic of Iran3 Tabriz University of Medical Sciences, Orthopedic Surgery Department,Tabriz, Islamic Republic of Iran4 Islamic Azad University, Medical Faculty, Tabriz, Islamic Republic of IranIntroduction: One of the most important concerns in hospitals isantimicrobial resistance in hospital pathogens that puts patients in risk ofmorbidity and mortality. It is caused by plasmid mediated resistanceagainst beta lactams by producing extended spectrum beta lactamasesenzyme (ESBLs). Pseudomonas aeruginosa is one of those pathogens thatis in common with most of these nasocomial infections. This study wasconducted to find the role of Pseudomonas aeruginosa in nasocomialinfections to find the best approach for its treatment.Methods: Different Samples from different parts of like tracheal aspirate,urine, blood, bronchial aspirate, sputum, CSF, wound discharge, bonemarrow and peritoneal fluid of ICU patients of 5 hospitals in Tabriz weretaken. They were tested for susceptibility by Disk agar diffusion methodand screening of ESBL-producing by Double disc approximation test,respectively, also combined test disc method and MIC determination by E-test were adopted for confirmation. Extracting Plasmid DNA by Kado andLiu technique, the presences of bla CTX-M1, bla CTX-M2 were studied byPolymerase Chain Reaction (PCR).Results:248 ICU patients, infected by Gram-negative bacilli were studied.Pseudomonas aeruginosa was the second agents in nosocomial infection,67 (27%). The susceptibility test showed 29%, 38%, 53.9%, 98%, 96%,70% 100% and 100% resistance against Piperacillin, ceftazidim,Ofloxacin, Sulphamethoazle, Cefotaxime, ceftriaxone, Tetracycline andcefuroxime. The Double Disk Test showed 96.7%, 100% and 96.6%resistance against Ceftriaxone, Cefotaxime, and Ceftazidime. Thecombined Test showed 63.4% negative result against Cefotaxime andCefotaxime / Clavulanic acid and 67.5% against Ceftazidime andCeftazidime / Clavulanic acid. By E-test ESBLs production was detectedin P.aeruginosa(78%). In plasmid extraction 64.5 % of isolates harbored asingle plasmid of 63kb.On the basis of PCR results, All of strains lackedeither CTX-M-1 or CTX-M-2 gene to confirm the rule for bla CTX-M.Conclusion:Pesudomonas aeruginsawas one of the most prevalent bacteria.Highest rate of resistance was showed against Cefuroxime, Tetracyclineand lowest rate was showed against Amikacin and Piperacillin. Our resultsshowed that DDT test was not as sensitive as CT and MIC methods and nostatistical significant difference was found between results of CT and MIC.Confirming no rules for suspicious genes by PCR, 78% of strains werefounded as ESBL producer. Since the genes encoding theses enzymes aremainly located on plasmids, so transmission of the plasmids coulddisseminate the resistance in future, unless the consumption ofcephalosporins are restricted and antibiotics such as imipenem substitudedfor the third generation cephalosporins, because these antibiotics,especially ceftazidim and ceftriaxone are strong inducers of ESBLs.HMP005Identification of D-tryptophan as immunologically activecompound excreted by probiotic bacteria using immunological invitro-test systemsI. Kepert 1,2 , J. Fonseca 3 , K. Hochwind* 4 , S. van Hemert 5 , M. Schmid 4 ,P. Schmitt-Kopplin 3 ,S. Krauss-Etschmann 1,2 ,A. Hartmann 41 Dr.von Haunersches Kinderspital ,Munich ,Germany2 Helmholtz Zentrum, Comprehensive Pneumology Center, Munich, Germany3 Helmholtz Zentrum, Research Unit BioGeoChemistry and Analytics, Munich,Germany4 Helmholtz Zentrum, Department Microbe-Plant Interactions, Munich, Germany5 Winclove, Amsterdam, NetherlandsThe interest of probiotic bacteria in health care is increasing in the lastyears. However there is still a lack of understanding the underlyingmechanisms. One reason for the detected health benefits might be thecrosstalk between bacteria and the host which is difficult to predict.Therefore, we aimed to identify soluble compounds produced by probioticbacteria using in vitro screening tools, high performance chemical analysisand metabolic profiling.Gram-positive probiotic bacteria were grown in defined minimal mediumand supernatants were taken at stationary phase. Culture supernatants,separated fractions and pure compounds were screened for their ability toprevent the lipopolysaccharide-induced maturation of human monocytederiveddendritic cells (DC). The activation markers CD83, CD86, CD80,and CD40 were measured with flow cytometry. Furthermore tests wereperformed indicating inhibition of the allergy-related thymus andactivation regulated chemokine (TARC) secretion of KM-H2 HodgkinLymphoma cells (ELISA). Immune-active supernatants were fractionatedby solid phase extraction and elution was performed with increasingmethanol concentration in water. After separation immune-active fractionsand compounds were analysed using FTICR-MS and NMR.Supernatants of several bacterial strains significantly down-regulated theexpressing of the activation markers CD83, CD86 and CD40. They alsosignificantly reduced TARC secretion of KM-H2 cells. After fractionationof two selected supernatants, the immune modulatory activity was found inthe 20%, 40% and 50% methanol fractions. We focused on the compoundin the 20% methanol fraction and were able to characterize it as D-tryptophan. In addition it was the only D-amino acid having this immunemodulatory effect when we tested the pure D-amino acids.The present work demonstrated that small extracellular molecules fromprobiotic strains have immune modulatory activity in the screeningsystems applied. Our work provides evidence that D-tryptophan is one ofseveral hitherto not yet described small molecules of probiotic bacteriawhich potentially interfere with human immune responses. This may givefurther basis for the application of these compounds as food additives tofinally provide anti-allergic effects.HMP006Metabolic characterization of dental biofilms produced byStreptococcus mutans under dietary carbohydrate exposureE.-M. Decker*, G. Maier, C. von OhleUniversity Centre of Dentistry, Oral Medicine and Maxillofacial Surgery,Department of Operative Dentistry and Periodontology, Tübingen, GermanyStrep. mutans is known to be a major pathogen for dental caries andhuman tooth decay. Especially, sucrose induces high cariogenicity byactivating microbial glycosyltransferases and thus production ofextracellular adhesive polysaccharides. The aim of the study was themetabolic characterization of Strep. mutans biofilm formation in thepresence of glucose, sucrose and the sugar alcohol xylitol. Biofilms ofStrep. mutans were produced metabolizing Schaedler-broth (0.58%glucose) (M1), Schaedler-broth with 5% sucrose as supplement (M2) andSchaedler-broth wih 1% xylitol in addition (M3) on human enamel slides.After 24h incubation time at 37°C the surface-associated biofilms werelabeled using three specific staining protocols: 1. live/dead celldifferentiation, 2. staining of total bacterial cells and extracellularpolysaccharides (EPS) by concanavalin A and 3. staining of total bacterialcells and microbial respiratory activity. The marked biofilms wereanalysed by confocal laser scanning microscopy and checked for microbialtotal cell counts and colony growth on Schaedler agar plates. Ten series ofeach approach were performed and analysed by means of one-way analysisof variance and Tukey Kramer statistical tests. The streptococcal biofilmthickness and volume reached its maximum under sucrose exposition inM2. Regarding the stratification of the biofilms the ConA-based EPSsignalsin all three media (M1-M3) showed higher acitivity in the internalregions of biofilms near to enamel compared with outer biofilm regions.The microbial respiratory activity tended to be lower in M2 in comparisonwith M1 and M3. In the presence of sucrose Strep. mutans biofilmsappeared as microcolonies associated with increased viability parameterslike biofilm depth, volume and higher vitality proportions compared to thecorresponding biofilms grown in media with glucose or xylitol. Within theBIOspektrum | Tagungsband 2012