First Case of High-Level Azithromycin-Resistant Neisseria gonorrhoeae in North Carolina.

We report on the first high-level azithromycin-resistant Neisseria gonorrhoeae isolate (minimum inhibitory concentration, ≥256 µg/mL) in North Carolina isolated from a pharyngeal swab of a 33-year-old HIV-negative man who has sex with men. In addition, the isolate was found to be susceptible to cefixime, ceftriaxone, and penicillin and resistant to tetracycline. By whole-genome sequencing, the strain was assigned as MLST ST9363, NG-MAST ST5035, and a novel NG-STAR sequence type, ST1993.

Rapid Identification of Klebsiella pneumoniae by Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry and Detection of Meropenem Resistance by Flow Cytometric Assay.

BACKGROUND: The aim of this study was to develop a rapid detection method of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains both MALDI-TOF MS and flow cytometry (FCM). METHODS: A total of 174 K. pneumoniae strains were included in this study. Molecular characterization of carbapenemase gene was performed by PCR. Bacterial identification was performed by MALDI-TOF-MS. Meropenem susceptibility was tested at the concentrations of breakpoints described by the Clinical and Laboratory Standards Institute (CLSI) guide by FCM. RESULTS: Sixty-two CRKP were positive for at least one carbapenemase gene. A total of 174 K. pneumoniae isolates obtained from clinically relevant material were correctly identified by Bruker MALDI-TOF MS with log (score) >2.0. These results were 100% concordant with the Phoenix™ Automated Microbiology System (BD, MD) and conventional identification results. Based on the analysis of the receiver operating characteristic (ROC) curves, the best validity and sensitivity data were obtained with a cut-off value of 18.88% by FCM. The concordance, sensitivity, and specificity for FCM by the selected cut-off values were 99.4%, 98.9%, and 100%, respectively. CONCLUSIONS: We conclude that reliable results on bacterial identification and meropenem susceptibility test can be obtained within 2 hr combined by MALDI-TOF-MS and FCM.

[Investigation of carbapenemases in carbapenem-resistant Escherichia coli and Klebsiella pneumoniae strains isolated in 2014 in Turkey]. / Türkiye'de 2014 yili içinde izole edilen karbapeneme dirençli Escherichia coli ve Klebsiella pneumoniae izolatlarinda karbapenemaz varliginin arastirilmasi.

Carbapenems are the choice of treatment in infections caused by multidrug resistant Enterobacteriaceae. In recent years carbapenem-resistant Enterobacteriaceae isolates due to carbapenemases have been increasingly reported worldwide. Multicenter studies on carbapenemases are scarce in Turkey. The aim of this study was to determine the distribution of carbapenemases from different parts of Turkey as a part of the European Survey of Carbapenemase Producing Enterobacteriaceae (EuSCAPE) project. Beginning in November 2013, carbapenem-resistant isolates resistant to at least one of the agents, namely imipenem, meropenem, and ertapenem were sent to the coordinating center. Minimum inhibitory concentrations for these carbapenems were determined by microdilution tests following EUCAST guidelines. Production of carbapenemase was confirmed by combination disk synergy tests. Types of carbapenemases were investigated using specific primers for VIM, IMP; NDM, KPC and OXA-48 genes by multiplex polymerase chain reaction. In a six month period, 155 suspected carbapenemase-positive isolates were sent to the coordinating center of which 21 (13.5%) were E.coli and 134 (86.5%) were K.pneumoniae. Nineteen (90.5%) strains among E.coli and 124 (92.5%) strains among K.pneumoniae were shown to harbour at least one carbapenemase gene by molecular tests, with a total of 92.3% (143/155). Carbapenemases were determined as a single enzyme in 136 strains (OXA-48: 84.6%; NDM: 6.3%; VIM: 2.8%; IMP: 1.4%) and as a combination in seven isolates (OXA-48 + NDM: 2.1%; OXA-48 + VIM: 2.1%; VIM + NDM: 0.7%). KPC was not detected in any of the isolates. According to the microdilution test results, resistance to imipenem, meropenem and ertapenem in OXA-48 isolates were 59.5%, 52.9% and 100%, respectively. The combination disk synergy test was 100% compatible with the molecular test results. As most of the OXA-48 producing isolates were susceptible to meropenem but all were resistant to ertapenem, ertapenem seems to be the most sensitive agent in screening carbapenemases in areas where OXA-48 is prevalent and phenotypic combination tests can be useful in centers where molecular tests are not available.

Effect of antibiotic use on bacterial flora of tonsil core in patients with recurrent tonsillitis.

The aim of this study was to investigate the effects of commonly used antibiotics on bacterial flora of the tonsil core. Patients who underwent tonsillectomy for recurrent chronic tonsillitis were included in the study. Three groups were formed: group 1 was treated for 10 days preoperatively with amoxicillin/clavulanic acid; group 2 was treated for 10 days preoperatively with clarithromycin; and group 3 included patients who underwent tonsillectomy without preoperative antibiotic use. The removed palatine tonsils were sent to our microbiology department in sterile tubes for bacteriological analysis. Seventy-three patients (group 1 = 19, group 2 = 20, group 3 = 34 patients) aged 3-18 years (mean 7 years) were included in the study. At least one bacterium was isolated from all tonsils, except for two cases in group 1; the difference in single bacterial growth among groups was not significant (p = 0.06). On the other hand, the numbers of patients with pathogenic bacterial growth was significantly lower in group 2 (n = 2) compared with group 1 (n = 10) and group 3 (n = 27) (p < 0.001). The bacterium isolated most frequently from the tonsils was Streptococcus viridans. Pseudomonas aeruginosa was the only pathogenic bacterium that grew in all three groups. Clarithromycin was more effective than amoxicillin/clavulanic acid in eradicating pathogenic bacteria in the tonsil core. Pseudomonas aeruginosa might be responsible for resistant or recurrent tonsil infections. To prevent endocarditis, antibiotic prophylaxis toward S. viridians, which is the most prevalent bacterium in the tonsil core, should be kept in mind for patients with heart valve damage.

[Investigation of the presence of mecC and Panton-Valentine leukocidin genes in Staphylococcus aureus strains isolated from clinical specimens during seven years period]. / Yedi yillik sürede klinik örneklerden izole edilen Staphylococcus aureus suslarinda mecC ve Panton-Valentine lökosidin gen varliginin arastirilmasi.

Detection and identification of methicillin-resistant Staphylococcus aureus (MRSA) in clinical microbiology laboratories are important for the selection of appropriate treatment and obtaining epidemiological data. mecC gene, is a mecA homologue, showing almost 69% DNA similarity with the mecA gene and the encoded protein by this gene shows almost 63% similarity with the PBP2a/2' protein. Several studies indicated that mecC positive MRSA strains can be transmitted from the livestock to humans by cross contamination. Panton-Valentine leukocidin (PVL), a potent cytotoxin of S.aureus is also considered as an important virulence factor. The aim of this study was to determine the existence and prevalence of mecC and pvl genes among S.aureus strains isolated from clinical specimens. A total of 1700 S.aureus isolates including 1177 methicillin-susceptible S.aureus (MSSA) and 523 MRSA, isolated in our hospital between January 2007 to December 2014, were included in the study. The isolates were identified by both conventional methods and BD Phoenix automated system (BD Diagnostic Instrument Systems, USA). Antibiotic susceptibility testing was performed by Kirby-Bauer disk diffusion method with oxacillin (1 µg) and cefoxitin (30 µg) according to the CLSI standards. The presence of mecA gene was investigated by the use of real-time PCR, and the presence of pvl and mecC genes were detected by conventional PCR method. Among the patients, 44.6% (759/1700) were outpatients, 65.8% (1119/1700) were male and the mean age of of patients was 39.7 years. Of 1700 isolates evaluated in this study, 523 (30.7%) were positive for mecA gene, however all of them were negative for mecC gene. A total of 32 (1.8%) isolates were positive for pvl gene including 23 (1.9%) out of 1177 MSSA and nine (1.7%) out of 523 MRSA strains. Eighteen (56.2%) of the PVL-positive S.aureus strains were isolated from skin and soft tissue infections. The frequency of PVL detected in this study was similar to the data of previous studies in our country. To the best of our knowledge, this is the first study for the determination of the mecC in our country. Although the mecC gene positive S.aureus has not been detected in our study, it should be kept in mind that the regional epidemiological conditions can vary quickly. In conclusion, multicenter studies are needed for the screening of mecC gene including the animal isolates, in Turkey.

[Rapid detection of extended-spectrum beta-lactamases by flow cytometry method]. / Genislemis spektrumlu beta-laktamazlarin akim sitometrisi yöntemiyle hizli tespiti.

Extended-spectrum beta-lactamases (ESBL), produced by Enterobacteriaceae members are enzymes that especially cause a resistance to cephalosporin group antibiotics commonly used in clinics. Early and rapid detection of ESBL production is crucial for antimicrobial treatment and infection control; however the methods used for this purpose are time consuming (24 to 48 hours). The aim of this study was to determine a flow cytometry based-test which provides to detect ESBL producing bacteria in a short time. A total of 38 ESBL-producing (29 Escherichia coli, 9 Klebsiella pneumoniae) and 10 non-producing (5 E.coli, 5 K.pneumoniae) Enterobacteriaceae strains isolated between 2012 and 2013 were included in this study. The identification and antibiotic susceptibility tests of the isolates were performed by using Phoenix(TM) 100 automated system (Becton Dickinson, USA). The presence of bla(TEM), bla(SHV), bla(CTX-M1), bla(CTX-M2) and bla(CTX-M9) genes were investigated in ESBL positive isolates via polymerase chain reaction method. At least one of the ESBL genes were detected in 36 out of 38 isolates and no genes were detected in two E.coli isolates. In flow cytometric method, the percentages of death cells exposed to cephalosporin [(ceftazidime (CAZ) or cefotaxime (CTX)] and clavulanic acid (CLA) combination, were compared with death cells exposed only to cephalosporin (CAZ or CTX). CLA index values (CAZ-CLA and CTX-CLA indices) were obtained for CTX and CAZ. Index values which was higher than 1.5 just for one cephalosporin were accepted as GSBL positive. The mean index values for CTX-CLA in ESBL positive strains according to their genotypic characteristics were between 1.14 and 7.22, while those values for CAZ-CLA were between 0.85 and 5.6. When the two groups of 38 ESBL positive and 10 ESBL negative strains were evaluated, statistically significant difference was detected for both CAZ-CLA and CTX-CLA indices (p< 0.005). CTX-CLA indices (p= 0.001) shown a better determination of ESBL when CAZ-CLA and CTX-CLA indices were compared statistically. In conclusion, flow cytometry is a rapid and reliable method for the detection of ESBL in clinical microbiology laboratories when compared with the other methods.

FKS mutant Candida glabrata: risk factors and outcomes in patients with candidemia.

BACKGROUND: Echinocandins are recommended for Candia glabrata candidemia. Mutations in the FKS1 and FKS2 genes are associated with echinocandin resistance. Few studies have assessed risk factors for FKS mutant isolates and outcomes in patients receiving micafungin treatment. METHODS: Patients with C. glabrata bloodstream infection admitted to a large, tertiary care hospital between 2009 and 2012 were included in this study. For each isolate, FKS1 and FKS2 genes were sequenced to identify mutations. Risk factors for FKS mutations and treatment outcomes in patients receiving an echinocandin were assessed using multivariate logistic regression. RESULTS: Seventy-two patients were included in the study of which 13 (18%) had an FKS mutant isolate. The only significant predictor for FKS mutations was prior echinocandin exposure (odds ratio [OR], 19.9; 95% confidence interval [CI], 4.7-84.7; P ≤ .01). Treatment failure occurred in 17 (30%) of 57 patients who received an echinocandin and was more common in patients with FKS mutants (6 of 10; 60%) compared with non-FKS mutants (11 of 47; 23%). Underlying gastrointestinal disorder (OR, 4.7; 95% CI, 1.1-20.9; P = .04) and prior echinocandin exposure (OR, 8.3; 95% CI, 1.7-40.4; P ≤ .01) were independent predictors of echinocandin treatment failure. Treatment response and echinocandin minimum inhibitory concentrations varied among specific FKS mutations. CONCLUSIONS: FKS mutations were identified in 18% of 72 patients with C. glabrata candidemia. Common risk factors for FKS mutant isolates included previous echinocandin exposure, which also influenced response rates.

Effect of VEGF gene therapy and hyaluronic acid film sheath on peripheral nerve regeneration.

Functional nerve regeneration after reconstructive nerve surgery remains unsatisfying. In this study, vascular endothelial growth factor (VEGF) gene therapy combined with a hyaluronic acid (HA)-enriched microenvironment in nerve regeneration was investigated. Sciatic nerve was transected, and end-to-end neurorrhaphy was performed on 32 male Sprague-Dawley rats, which were randomly divided into four groups (n = 8 per group): nerve coaptation without treatment (group I); nerve coaptation covered with HA film sheath (group II); nerve coaptation with intramuscular VEGF gene in plasmid injection (group III); and nerve coaptation combined with HA film sheath and intramuscular VEGF gene in plasmid injection (group IV). Contralateral sciatic nerves were used as control. VEGF expression was verified from gluteal muscle biopsies surrounding the sciatic nerve by reverse transcriptase-PCR. Electrophysiological, histopathological, and electron microscopic evaluations were performed after 4 weeks. Mean peak amplitude of groups I-IV and nonoperated sciatic nerve were 4.5 ± 0.6 mV, 6.4 ± 0.4 mV, 6.7 ± 0.5 mV, 8.5 ± 0.4 mV, and 9.8 ± 0.5 mV, respectively. Mean myelinated axonal counts of groups I-IV and nonoperated sciatic nerve were 105 ± 24, 165 ± 19, 181 ± 22, 271 ± 23, and 344 ± 17, respectively. Treatment with HA film sheath coverage combined with intramuscular VEGF gene in plasmid injection yielded statistically significant higher peak amplitudes and myelinated axonal counts (P < 0.001). In addition, significantly less scar formation with HA administration (groups II and IV; P < 0.001) was found. Thus, it was found that VEGF might crucially regulate nerve regeneration processes and that HA can reduce the scar formation. This study showed that the combination of HA film sheath and VEGF gene may synergistically promote peripheral nerve regeneration.

[Identification of staphylococci directly from positive blood culture bottles by MALDI-TOF MS system]. / Stafilokoklarin pozitif kan kültür siselerinden MALDI-TOF MS sistemi ile direkt olarak tanimlanmasi

Bloodstream infections are substantial causes of morbidity and mortality worldwide. Staphylococcus species are the most commonly isolated microorganisms from blood cultures in clinical microbiology laboratories. MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time- of- Flight Mass Spectrometry) system allows the identification of microorganisms directly from positive blood culture bottles. The aim of this study was to evaluate the MALDI-TOF MS system for the identification of staphylococci directly from the positive blood culture bottles which revealed the presence of gram-positive cocci by staining. A total of 96 positive blood culture bottles that yielded gram-positive cocci by Gram stain were evaluated. These blood cultures were obtained from 69 patients between December 2013-February 2014. Conventional methods and BD Phoenix™ automated bacterial identification system (Becton Dickinson, USA) were used for routine identification. The strains were also identified by real-time Taqman PCR (qPCR) which was considered as the reference method. In MALDI-TOF MS method, MALDI Sepsityper™ Kit was used for the bacterial identification and all measurements were carried out by using Microflex LT instrument and FlexControl 3.0 software (Bruker Daltonics, USA). Of 96 culture bottles positive for gram-positive cocci, 90 were correctly identified as staphylococci at genus level with all the three study methods (qPCR, BD Phoenix, Bruker MALDI-TOF MS). The other six samples were identified as Enterococcus faecium (n= 4) and Streptococcus pyogenes (n= 2) by both Phoenix and the MALDI-TOF systems. Of the 90 samples, 87 were identified at the species level (15 S.aureus, 33 S.epidermidis, 29 S.hominis, 10 S.haemolyticus) and three at the genus level by the reference qPCR method. When comparing the results obtained by qPCR and Bruker MALDI-TOF MS, incompatibility was detected for three isolates. Those isolates were identified as S.hominis by qPCR, however two of them were identified as S.haemolyticus and one as S.epidermidis by MALDI-TOF MS. Compared with real-time Taqman PCR it was detected that Bruker MALDI TOF MS was identified 100% of S.aureus to the genus and species level and 100% and 96.6% of coagulase-negative staphylococci (CNS) to the genus and species level, respectively. In conclusion, it was thought that Bruker MALDI TOF MS system may allow rapid and reliable identification of S.aureus and CNS directly from positive blood culture bottles compared with the routine methods used in the clinical microbiology laboratory.

[Clostridium difficile infection: epidemiology, risk factors, pathogenesis, clinical features, diagnosis and therapy]. / Clostridium difficile enfeksiyonu: Epidemiyoloji, risk faktörleri, patogenez, klinik özellikler, tani ve tedavi.

Clostridium difficile is a gram-positive, spore-forming anaerobic bacterium. C.difficile is the leading cause of antibiotic associated diarrhea and colitis. The clinical spectrum of C.difficile infection (CDI) is highly variable, ranging from mild diarrhea to severe forms of intestinal illness including toxic megacolon, ileus, bowel perforation, and pseudomembranous colitis. Advanced age, long duration of hospitalization, and exposure to certain antimicrobial agents are the most common risk factors for CDI. The main virulence determinants of C.difficile are toxin A (enterotoxin) and toxin B (cytotoxin). Diagnosis of CDI is based on the identification of C.difficile toxin A or B in diarrheal stool. Various laboratory tests have been currently developed for the detection of C.difficile or its toxins in stool samples. The cell culture cytotoxicity assay and toxigenic culture have been regarded as the reference standard methods for the detection of C.difficile toxins. However, both of the reference methods are laborious, time consuming, and need expert personnel. Therefore, many microbiology laboratories use enzyme immunoassays, glutamate dehydrogenase antigen tests and real-time polymerase chain reaction (PCR) which are more rapid and practical. First-line antibiotics for CDI treatment are metronidazole and vancomycin. In this review, epidemiology, clinical spectrum, risk factors, pathogenesis, diagnostic methods and treatment of CDI have been summarized.

In vitro evaluation of the minimum bactericidal concentrations of different root-end filling materials.

AIM: The aim of this study was to evaluate the minimum bactericidal concentrations (MBC) of root-end filling materials ProRoot MTA, MTA Angelus and IRM. MATERIALS AND METHODS: Macrodilution broth method was used. Microorganisms used were: Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212) and Streptococcus mutans. Serial two-fold dilutions of root-end filling samples were prepared in macrodilution tubes with concentrations ranging from 1/2 to 1/512. The samples dilutions were incubated for 24 hours. After incubation, 0.1 ml of diluted culture was inoculated onto the surface of supplemented sheep blood agar (Merck, Germany) and all plates were incubated at 37°C in aerobic condition for 24 hours. The MBC was defined as the lowest concentration of root-end filling samples where no growth was recorded. RESULTS: MBC of both mineral trioxide aggregate (MTA) products against S. aureus were recorded as 15.62 mg/ml and for IRM 31.25 mg/ml MBC for both MTA groups against E. faecalis were recorded as 31.25 mg/ml and for IRM 62.5 mg/ml. MBC of all root-end filling samples against S. mutans were recorded as 62.5 mg/ml. CONCLUSION: All tested root-end filling materials showed acceptable MBC against S. aureus and E. faecalis. CLINICAL SIGNIFICANCE: All tested materials can be used safely for filling of a root-end cavity.

In Vitro Activity of Omadacycline and Comparator Antibiotics against Extended-Spectrum Beta-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae Urinary Isolates.

Limited oral antibiotic options exist for urinary tract infections (UTI) caused by ESBL-producing Enterobacterales. The aim of the study was to evaluate in vitro activity of omadacycline and comparator antibiotics against clinical ESBL-producing and non-ESBL-producing E. coli and K. pneumoniae urinary isolates. 102 isolates each of E. coli and K. pneumoniae were collected from clinical urine specimens in 2019. By design, an equal number of each species were included that tested positive and negative for ESBL production. Omadacycline MICs were determined using gradient test strips and compared to MICs of comparator antibiotics as determined by an automated broth microdilution system. Isolates were considered susceptible to omadacycline if the MIC was ≤4 µg/mL for each species. 54.9% of all ESBL-producing isolates were susceptible to omadacycline, but better susceptibility was observed for ESBL-producing E. coli (74.5%). Omadacycline MICs were 2-4 fold lower for E. coli and K. pneumoniae strains not producing ESBL. The omadacycline MIC 50 and 90 values were 4 and 16 µg/mL, respectively, for all isolates studied. 74.5% of all isolates were considered susceptible to omadacycline. MICs were generally lower for E. coli strains with MIC 50 and 90 values of 4 and 8 µg/mL, respectively (87.3% susceptible), compared with K. pneumoniae. Overall, the most active agents were omadacycline and nitrofurantoin, while other comparator antibiotics were less active. Omadacycline represents a promising oral antibiotic for treating UTI caused by ESBL-producing E. coli, particularly when resistance limits other oral options. Prospective, controlled clinical trials are needed to validate these in vitro results.

Clinical impact of a multiplex rapid diagnostic pneumonia panel in critically ill patients.

Objective: To evaluate the clinical impact of the BioFire FilmArray Pneumonia Panel (PNA panel) in critically ill patients. Design: Single-center, preintervention and postintervention retrospective cohort study. Setting: Tertiary-care academic medical center. Patients: Adult ICU patients. Methods: Patients with quantitative bacterial cultures obtained by bronchoalveolar lavage or tracheal aspirate either before (January-March 2021, preintervention period) or after (January-March 2022, postintervention period) implementation of the PNA panel were randomly screened until 25 patients per study month (75 in each cohort) who met the study criteria were included. Antibiotic use from the day of culture collection through day 5 was compared. Results: The primary outcome of median time to first antibiotic change based on microbiologic data was 50 hours before the intervention versus 21 hours after the intervention (P = .0006). Also, 56 postintervention regimens (75%) were eligible for change based on PNA panel results; actual change occurred in 30 regimens (54%). Median antibiotic days of therapy (DOTs) were 8 before the intervention versus 6 after the intervention (P = .07). For the patients with antibiotic changes made based on PNA panel results, the median time to first antibiotic change was 10 hours. For patients who were initially on inadequate therapy, time to adequate therapy was 67 hours before the intervention versus 37 hours after the intervention (P = .27). Conclusions: The PNA panel was associated with decreased time to first antibiotic change and fewer antibiotic DOTs. Its impact may have been larger if a higher percentage of potential antibiotic changes had been implemented. The PNA panel is a promising tool to enhance antibiotic stewardship.

[Investigation of various virulence factors among the hospital and community-acquired Staphylococcus aureus isolates by real-time PCR method]. / Hastane ve Toplum Kaynakli Staphylococcus aureus Izolatlarinda Çesitli Virülans Faktörlerinin Gerçek Zamanli PCR Yöntemiyle Arastirilmasi

Staphylococcus aureus is the most common cause of skin and soft tissue infections in the community and the most important cause of nosocomial infections. In this research, it was aimed to detect the presence of staphylococcal enterotoxin A to D (SEA, SEB, SEC and SED, respectively), toxic shock syndrome toxin-1 (TSST-1), Panton-Valentine leukocidin (PVL) and SCCmec phenotype in methicillin-resistant S.aureus (MRSA) isolates and to demonstrate the genotypic association between hospital acquired and community acquired isolates. In the study the virulence factors of 147 S.aureus strains isolated from various clinical samples at Gulhane Military Medical Academy Hospital Microbiology Laboratory between 2007 and 2010 were investigated by real-time polymerase chain reaction. MLVA (multiple locus variable number of tandem repeat analysis) method was used to demonstrate the genotypic association between hospital-and community-acquired isolates. Seventy-two (%48.9) of 147 S.aureus isolates were determined as community acquired and 75 (%51.1) as hospital acquired. Ninety-three (63.2%) isolates possesed at least one toxin (77 strains harboured one, and 16 strains harboured more than one). Of the isolates in which toxin was detected 59.1% (55/93) were hospital acquired, 40.9% (38/93) were community acquired. SEA was determined in 59 (40.1%), SEB in 8 (5.4%), SEC in 12 (8.1%), SED in 8 (5.4%), TSST- 1 in 17 (11.5%) and PVL in 6 (4.0%) of the isolates. Methicillin resistance was determined in 61.1% (44/72) of the hospital acquired isolates and 6.7% of the (5/75) community acquired isolates. In our study, SCCmec type III was detected in 90.9% (40/44) of hospital acquired MRSA isolates and SCCmec type IV in 40.0% (2/5) of community acquired MRSA isolates. Most of the strains (40/47; 85.1%) carrying SEA were hospital acquired, and they were determined as methicillin-resistant. According to MLVA, hospital and community acquired groups' clustering rates, number of clones, number of unique profile were determined as; 73.6% and 57.3%; 34% and 47%; 19% and 32%, respectively. It was concluded that high prevalence of SEA toxin in hospital acquired MRSA isolates indicated that there was a possible association between the presence of toxin and antimicrobial resistance.

Evaluation of the Potential Impact of a Multiplex Rapid Diagnostic Panel in Critically Ill Patients With Hospital-Acquired Pneumonia.

Background Rapid diagnostic tools have emerged as valuable assets assisting clinicians in decision-making regarding patient management in the hospital setting. Our study sought to identify the potential impact of the BioFire® FilmArray® Pneumonia Panel (FP Panel) (BioFire Diagnostics, Salt Lake City, UT, USA) in patients with hospital-acquired pneumonia (HAP). Methods Respiratory samples obtained by bronchoalveolar lavage (BAL) or tracheal aspiration (TA) from ICU patients with a diagnosis of HAP were tested by the FP panel in addition to routine bacterial cultures. In addition, the electronic health records of these patients were reviewed to determine what potential changes in antimicrobial therapy could have been implemented if the panel results were known to the treatment team in real-time. A cost analysis was also performed incorporating the cost of the pneumonia panel and the savings associated with the potential decrease of antibiotic use and avoidance of the rapid viral diagnostic panel.  Results Fifty-six patients met the study criteria. The FP panel results could have prompted a change in therapy in 36 (64.3%) patients, with an anticipated mean reduction in time to optimized therapy of approximately 51 hours. In addition, the panel identified three cases where antimicrobials should have been altered because patients were not receiving empiric therapy with activity against the causative pathogen and 34 opportunities for antibiotic de-escalation. The cost analysis calculated an additional cost of $10 per patient associated with using the FP panel.  Conclusions The FP panel could have prompted a change in therapy in about two-thirds of patients studied. Its potential benefits include a more rapid time to optimized therapy, reduced exposure to and cost of broad-spectrum antimicrobials, and reduced cost of other rapid diagnostic tests.

Evaluation and comparison of the effects of hyperbaric oxygen and ozonized oxygen as adjuvant treatments in an experimental osteomyelitis model.

BACKGROUND: We evaluated and compared the efficacy of ozone (O(3)) and hyperbaric oxygen (HBO) therapies in an experimental rat model of osteomyelitis. MATERIALS AND METHODS: Forty-eight male Sprague-Dawley rats were divided into sham, osteomyelitis (control), vancomycin (V), vancomycin + HBO (VHB), vancomycin + O(3) (VO), and vancomycin + HBO + O(3) (VOHB) groups. Osteomyelitis was induced by a bone injection of 10(8) CFU/mL methicillin-resistant Staphylococcus aureus. HBO was administered daily at 2.8-atm pressure for 90 min; O(3) therapy was provided as intraperitoneal injections of 0.7 mg/kg O(3)/O(2) gas mixture once daily. Treatments were continued from d 7 to 21 after induction of osteomyelitis. Bone tissues and blood samples were harvested for biochemical, histopathologic, and microbiologic analyses. RESULTS: Rats in the sham, VO, and VOHB groups gained weight but those in the control, V, and VHB groups did not. Levels of malondialdehyde, superoxide dismutase, and glutathione peroxidase were lower in the VHB, VO, and VOHB groups than in V and control groups. Levels of interleukin-10 and -1ß and tumor necrosis factor-α were decreased in the VHB, VO, and VOHB groups; transforming growth factor-ß was increased in these groups compared with V and control groups (P ≤ 0.001). Bacteria counts in VOHB were significantly lower than those in group of V (P = 0.012). Histopathologic scores in group VO were significantly lower than those in group V (P = 0.046). CONCLUSIONS: O(3) was as effective as HBO in decreasing oxidative parameters and inflammatory cytokines. Rats in the VO and VOHB groups gained more weight than did the other groups. Bacteria counts were significantly decreased in group VOHB compared with the other groups. Histopathologic scores in group VO were significantly decreased compared with the other groups.

Effect of a self-etching adhesive containing an antibacterial monomer on clinical periodontal parameters and subgingival microbiologic composition in orthodontic patients.

INTRODUCTION: The aims of this study were to evaluate the effect of a self-etching adhesive system containing an antibacterial monomer on periodontal health and subgingival microbiologic composition in orthodontic patients and to compare it with a conventional adhesive system. METHODS: A split-mouth design was chosen, and 15 patients were included in the study. Brackets in contralateral quadrants were bonded with either a conventional adhesive system (control) or a self-etching adhesive system that contained an antibacterial monomer. Clinical periodontal parameters including plaque index, gingival index, probing depths, and bleeding on probing were determined. Subgingival plaque samples were collected before bracket placement (T0) and at the 6-month follow-up (T1). The real-time TaqMan polymerase chain reaction assay was used to determine the subgingival counts of Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Campylobacter rectus. For clinical periodontal parameters, analysis of covariance (ANCOVA) and, for bacterial counts, Wilcoxon tests were used for statistical comparisons at the P <0.05 level. RESULTS: Clinical periodontal parameters were not changed, and they were not different between the groups from T0 to T1. T forsythensis and F nucleatum increased during the treatment period in both groups (P <0.05). The majority of the bacteria were T nucleatum at T0 and T1 in both groups. Changes in bacterial load from T0 to T1 were not different between groups except for T forsythensis and F nucleatum (P <0.05). CONCLUSIONS: The use of an antibacterial monomer did not have an additional positive effect on clinical periodontal parameters. When used in bonding orthodontic brackets, the antibacterial monomer failed to reduce periodontopathogenic bacteria when compared with the conventional adhesive system during a 6-month treatment period.

Evaluation for Helicobacter pylori in Meckel's diverticulum by using real-time PCR.

BACKGROUND: Although Helicobacter pylori (H. pylori) has been identified in heterotopic gastric mucosa of Meckel's diverticulum, controversial results are reported in the pertinent literature. AIMS: The aim of this study was to evaluate for the presence of H. pylori histologically using hematoxylin-eosin and Toluidine Blue in Meckel's diverticulum and by real-time TaqMan polymerase chain reaction (PCR) in those with heterotopic gastric mucosa. METHODS: The study included 21 consecutive patients who had undergone resection of Meckel's diverticulum at our hospital between 1995 and 2007. The paraffin-embedded tissues were retrieved and reviewed for the presence of histological abnormalities and H. pylori-like organisms and for the presence or absence of heterotopic mucosa. H. pylori was sought in those cases that contained heterotopic gastric mucosa using real-time TaqMan PCR to amplify a fragment of the 23S ribosomal RNA (rRNA) gene of H. pylori. RESULTS: Upon histological examination, heterotopic gastric mucosa was found to be present in 12 cases. H. pylori was not identified in any of the sections examined. A genomic PCR product was also not obtained in real-time PCR study. CONCLUSIONS: We have confirmed that colonization of H. pylori, if it occurs at all, is exceedingly rare in heterotopic gastric mucosa of Meckel's diverticulum.

Simultaneous detection and differentiation of pathogenic and nonpathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay.

Leptospirosis, caused by pathogenic Leptospira, is one of the most important zoonoses in the world. Several molecular techniques have been developed for detection and differentiation between pathogenic and saprophytic Leptospira spp. The aim of this study was to develop a rapid and simple assay for specific detection and differentiation of pathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay using primers and probes targeting Leptospira genus specific 16S ribosomal RNA gene, the pathogen specific lig A/B genes and nonpathogen Leptospira biflexa specific 23S ribosomal RNA gene. Sixteen reference strains of Leptospira spp. including pathogenic and nonpathogenic and ten other negative control bacterial strains were used in the study. While the 16S primers amplified target from both pathogenic and non-pathogenic leptospires, the ligA/B and the 23S primers amplified target DNA from pathogenic and non-pathogenic leptospires, respectively. The multiplex real-time PCR (TaqMan) assay detection limit, that is, the sensitivity was found approximately 1 x 10(2) cells/ml for ligA/B gene and 23S ribosomal RNA gene, and 10 cells/ml 16S ribosomal RNA. The reaction efficiencies were 83-105% with decision coefficients of more than 0.99 in all multiplex assays. The multiplex real-time PCR (TaqMan) assay yielded negative results with the ten other control bacteria. In conclusion, the developed multiplex real-time PCR (TaqMan) assay is highly useful for early diagnosis and differentiation between pathogenic and non-pathogenic leptospires in a reaction tube as having high sensitivity and specificity.