Integron distribution and relationship to antimicrobial resistance in E. coli isolated from blood culture.

PURPOSE: The aim of this study was to evaluate the distribution of integrons in strains of E. coli isolated from blood culture and the relationship between integrons and antimicrobial resistance. METHODS: The study included 100 E. coli strains sent to the Medical Microbiology Laboratory from different clinics between September 2022 and June 2023. Antibiotic susceptibility was evaluated according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The presence of integrons was determined by the inhouse polymerase chain reaction (PCR). RESULTS: Integron positivity was detected in 45 (45%) of isolates, and class 1 integrons were found in 41 (41%), class 2 integrons in 2 (2%), and both class 1 integrons and class 2 integrons in 2 (2%). Class 3 integron positivity was not detected. In total, 63 cases of community origin and 37 cases of hospital origin were identified. When antibiotic resistance was evaluated, the highest sensitivity was noted for amikacin (1%), meropenem (5%), imipenem (6%), and the highest resistant antibiotics were ampicillin (82%), cepfuroxime sodium (65%), and amoxicillin/clavulanate (62%), respectively. Of the 16 antimicrobial substances evaluated, 10 had an antibiotic resistance rate of over 45%. In class 1 integron-positive samples, ampicillin resistance and trimethoprim/sulfamethoxazole resistance were higher than in negative samples (p = 0.02, p = 0.0001, respectively). Fifty-one (51%) samples were found to have multiple drug resistance (MDR). In total, 59.5% of hospital-acquired isolates and 46% of community-acquired isolates were considered to be MDR. The class 1 integron positivity in MDR samples was high (p = 0.038). CONCLUSION: The high MDR rates in both hospital-acquired and community-acquired isolates are alarming. In particular, class 1 integron monitoring is very important to prevent the spread of MDR isolates.

Is the Anal Component of the Anogenital HPV-Related Disease Overlooked During the Surveillance of Patients Treated for Cervical Intraepithelial Neoplasia?

AIM: To investigate the anal component of the anogenital Human Papillomavirus (HPV) related disease during surveillance of patients treated for cervical intraepithelial neoplasia (CIN). METHODS: Patients were analyzed within two groups according to the histopathological examination of the cervical biopsies: Low-Grade Squamous Intraepithelial Lesion (LSIL) and High-Grade Squamous Intraepithelial Lesion (HSIL) groups. Anal specimens were also collected in the first-year follow-up visit. RESULTS: All patients had cervical high-risk HPV (HR HPV) infection at admission. At the first-year follow-up, positive HR HPVs were found in 47% of cervical samples. Despite this clearance, the anal HPV infection rate after the first year was 42.5% and 39.6% in LSIL and HSIL groups. Amongst the HSIL group, anal HR HPV positivity was observed in 29.6% of cases without any cervical HPV infection. CONCLUSION: A group of women cured of high-grade lesions have ongoing anal HPV infection. It is reasonable to propose that detecting anal HPV could impact the patient's treatment process. Therefore, prospective studies are needed to investigate this group of women's clinical outcomes and define the clearance rate of cervical HPV infection when anal HPV persists.

A Decrease in Lactobacilli in the Vaginal Microbiota Is Independently Associated With HPV Persistence in Women With High-Risk HPV Infection.

Background Vaginal dysbiosis, an imbalance between species, can initiate some local changes in immune and metabolic signaling causing chronic inflammation. The mechanism of the clearance or progression of the HPV infection has not been uncovered yet. We hypothesized that vaginal dysbiosis may contribute to the persistence of the cervical HPV infection. Therefore we aimed to determine the association of lactobacillus dominancy index with cervical HR-HPV persistence. Methods A total of 100 women who were followed up because of high-risk HPV infection were defined as the target study group. The patients were evaluated in two groups; HPV positive (group with HPV persistence, n=43) and HPV negative (group with HPV clearance, n=57). Cervicovaginal swab samples and blood samples were evaluated for Nugent score, lactobacillus dominance, and white blood cell count. Statistical tests were performed by the IBM Statistical Product and Service Solutions (version 22, IBM SPSS Statistics for Windows, Armonk, NY) program. The continuous variables were presented using the mean±standard deviation (SD), and the categorical variables were presented as the number of cases and their percentage. A p value less than 0.05 (<0.05) was set as statistically significant. Results HPV persistence was observed in 43 (43%) patients. Univariate analysis revealed that age, menopausal status, and lactobacillus reduction were associated with HPV persistence (p<0.05). The median value of the Nugent score was similar among groups. After logistic regression analysis, lactobacillus reduction continued to be associated with HPV persistence, independent of age and menopause (OR: 2.668, 96% CI: 1.069-6.662, p<0.05) Conclusions A decrease in lactobacilli in the cervicovaginal microbiota is associated with the persistence of HPV, regardless of age and menopausal status in this study.

Evaluation of Viral and Bacterial Pathogens in the Central Nervous System Infections with Multiplex PCR.

OBJECTIVE: To evaluate the bacterial and viral causes of central nervous system (CNS) infection by multiplex PCR. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Medical Microbiology, Pamukkale University Faculty of Medicine, Turkey, from March 2016 to December 2021. METHODOLOGY: Cerebrospinal fluid (CSF) samples of patients prediagnosed with CNS infection were included in the study. Viral pathogens were detected with the Multiplex real-time PCR panel (FTD Neuro9, Fast Track Diagnostics, Luxembourg) and bacterial pathogens with the multiplex real-time PCR panel (FTD Bacterial Meningitis, Fast Track Diagnostics, Luxembourg). The identification of bacteria growing in samples was done by conventional methods and with the Phoenix™ (Becton Dickinson Diagnostics, USA) automated system. RESULTS: CSF samples of 440 patients were evaluated using multiplex PCR panel. The viral factors included adenovirus (14.2%), human herpes virus 7 (1.5%), varicella zoster virus (1.3%), herpes simplex virus 1 (1.3%), cytomegalovirus (1.3%), Epstein-Barr virus (0.8%), human herpes virus (0.8%), herpes simplex virus 2 (0.3%), varicella zoster virus (0.3%), and parvovirus B19 (0.3%); and bacterial factors included Streptococcus pneumoniae (7.0%) and Neisseria meningitidis (0.9%). The bacterial growth was detected in the CSF culture was 4.9%. Among the growing bacteria, there were six different types that were not found on the multiplex PCR panel. CONCLUSION: The use of a comprehensive bacterial multiplex PCR panel containing common pathogens will be more effective in pathogen detection. Care should be taken, especially when interpreting the viral Multiplex PCR. KEY WORDS: Viral multiplex PCR, Bacterial multiplex PCR, Bacteria culture.

Determination of COVID-19 PCR positivity and mutation status in coronaVac vaccinated individuals in Turkey.

BACKGROUND: CoronaVac is an inactivated virus-based COVID-19 vaccine used in Turkey and approved for emergency use by the World Health Organization (WHO). In this study, it was aimed to retrospectively evaluate the mutation status and clinical status in individuals who received two doses of CoronaVac vaccine and were infected with COVID-19 at least two weeks after the second dose. METHODS: 164 people were included in the study and COVID-19 diagnosis and mutation analyses were determined by RT-PCR using the Bioseepdy SARS CoV-2 Double Gene RT-qPCR Kit and the Biospeedy SARS-CoV-2 Variant Plus kit in accordance with the protocol determined by the manufacturer. RESULTS: 116 (70.7%) UK (Alpha, B.1.1.7) mutation and 3 (1.8%) South Africa (Beta, B.1.351), Brazil (Gamma, P.1) mutations were determined in 164 double doses CoronaVac vaccinated patients; 45 (27.5%) patients were mutation negative. Nine patients (5.5%) developed pneumonia. Eight patients (4.9%) had CT findings compatible with corona virus infection. Seven (4.3%) of the patients received treatment in the intensive care unit, and 5 (3%) of the patients were intubated. DISCUSSION: In conclusion, people who receive two doses of CoronaVac vaccine can be reinfected with mutant viruses, vaccine significantly reduces the need for hospitalization, CT findings and intensive care.

Rotavirus and adenovirus prevalence in patients with acute viral gastroenteritis in Denizli, Turkey, 2017-2021.

This study aims to determine retrospectively the prevalence of rotavirus and enteric adenovirus in patients with gastroenteritis symptoms and the distribution of pathogens by gender, age, seasons, and years. The stool samples sent to Pamukkale University Healthcare Research and Practice Hospital's Medical Microbiology laboratory between January 2017 and December 2021 were evaluated for rotavirus/adenovirus antigen positivity. Rotavirus and adenovirus antigens were studied with the Rotavirus-Adenovirus Combo Rapid Cassette Test (Acro Biotech) kit. Rotavirus was detected in 683 (8.2%) of the 8315 stool samples evaluated, and 180 (2.2%) samples were positive for adenovirus. Coinfection was detected in 21 (0.25%) samples. Rotavirus was found at the highest rate in 2019 (p = 0.001). The adenovirus was detected in 2020 at a lower rate than in other years (p = 0.0001). The rotavirus was observed at a higher rate in 0-<3, 3-<6, and 6-<13 age groups and adenovirus was detected at a higher rate in 3-<6 and 6-<13 age groups compared to other age groups (p = 0.001, p = 0.003, respectively). The highest rate of incidence of the rotavirus was found in spring and adenovirus in winter. In the etiology of gastroenteritis, especially in children, adenovirus and rotavirus should not be ignored in winter and spring. The prevalence of rotavirus was observed to have decreased in 2020 and onwards, and the prevalence of adenovirus decreased in 2020.

Monitoring SARS CoV-2 antibodies positivity in healthcare workers after inactivated CoronaVac® vaccine.

Introduction: In this study, we aimed to monitor anti-spike and anti-nucleocapsid antibodies positivity in healthcare workers (HCWs) vaccinated with two doses of inactivated CoronaVac® (Sinovac, China) vaccine. Methods: Overall, 242 volunteer HCWs were included. Of the participants, 193 were HCWs without history of prior documented COVID-19 (Group 1), while 49 had history of prior documented COVID-19 before vaccination (Group 2). The participants were followed up for SARS-CoV-2 antibodies positivity at four different blood sampling time points (immediately before the second vaccine dose and at the 1st, 3rd months and 141-150 days after the second dose). We investigated the serum IgG class antibodies against SARS-CoV-2 RBD region and IgG class antibodies against SARS-CoV-2 nucleocapsid antigen by chemiluminescent microparticle immunoassay (CMIA) method using commercial kits. Results: We found positive serum anti-RBD IgG antibody in 76.4% of the participants (71% in Group 1; 98% in Group 2) 28 days after the first dose. When the antibody levels of the groups were compared at the four blood sampling time points, Group 2 anti-RBD IgG levels were found to be significantly higher than those in Group 1 at all follow-up time points. Although anti-RBD IgG positivity persisted in 95.6% of all participants in the last blood sampling time point, a significant decrease was observed in antibody levels compared to the previous blood sampling time point. Anti-nucleocapsid IgG antibody was positive in 12 (6.2%) of participants in Group 1 and 32 (65.3%) in Group 2 at day 28 after the first dose. At the fourth blood sampling time point, anti-nucleocapsid antibodies were found to be positive in a total of 20 (9.7%) subjects, 10 (6.1%) in Group 1 and 10 (23.8%) in Group 2. Conclusions: In this study, it was determined that serum antibody levels decreased in both groups after the third month after the second dose in HCWs vaccinated with CoronaVac® vaccine.

Effects of xylitol impregnated toothbrushes on periodontal status and microbial flora in orthodontic patients.

OBJECTIVES: To investigate whether the use of xylitol-impregnated toothbrushes affects periodontal condition and microbial flora in orthodontic patients with poor oral hygiene. MATERIALS AND METHODS: Forty-four patients with baseline mean Turesky plaque index scores ≥1.5 were randomly divided into two groups. Half received xylitol-containing toothbrushes and the other half, xylitol-free toothbrushes. The periodontal measurements and saliva samples were taken at baseline (T0), 1 month later (T1), and 3 months after brushing (T2) to evaluate periodontal health and microflora changes. Periodontal status was assessed with plaque index (PI), gingival index (GI), and bleeding on probing (BOP) scores. Data were statistically analyzed with Mann-Whitney U and Friedman tests. RESULTS: All periodontal parameters significantly decreased from T0 to T1 and from T0 to T2 in both groups. The PI and GI scores reduced significantly in the control group, while BOP scores reduced in both groups between T1 and T2. Intergroup comparisons showed significant differences for BOP, PI, and GI at T0, T1, and T2 times, respectively. For microbial parameters, there were no statistically significant differences within groups from T0 to T1. Total bacterial counts significantly decreased in the xylitol group between T1 and T2. Decreases in Streptococcus mutans and total bacteria were significant in both groups from T0 to T2. No significant differences were found between the groups in microbial flora at any time. CONCLUSIONS: A 3-month use of xylitol-containing toothbrushes showed almost the same changes and provided no positive effects on periodontal and microbial parameters compared to the control group.

Predictors of Human papillomavirus (HPV) persistence after treatment of high grade cervical lesions; does cervical cytology have any prognostic value in primary HPV screening?

OBJECTIVE: This study aimed to determine the factors associated with Human Papillomavirus (HPV) persistence in women undergoing cervical excision for pre-invasive lesions, after they have been referred from a primary HPV screening program. METHODS: A retrospective study design involving patients who were treated at a Cervical Disease Screening and Treatment Unit, in a university hospital setting. After initial treatment, cervical HPV infection status was analyzed at the sixth month, first year and then subsequently after the second year. RESULTS: Totally, 395 patients who were diagnosed with pre-invasive cervical lesions and who subsequently undergone cervical excision were identified. In the first-year visit after cervical excision, HPV 18 was cleared in almost all (95.8%) cases, followed by HPV 16 (69.9%) and other hrHPV types (65.6%). Available data documented that 88.6% of women reached clearance after the two-year follow-up. Univariate analysis revealed a significantly higher proportion of HPV clearance among women who were younger (p = 0.019), premenopausal (p = 0.002), and who had been found to have a negative cytology result on their initial Pap test (p = 0.018). However, only cervical cytology result remained as the independent predictor of HPV persistence on a multivariate logistic regression (OR 0.43; 95% CI 0.21-0.87; p = 0.019). CONCLUSIONS: A low risk of HPV persistence was found among every HPV genotype in women undergoing cervical excision for pre-invasive cervical lesions. Initial cervical cytology result was the only independent predictor of HPV clearance during surveillance, which indicates the prognostic value of Pap test in primary HPV screening.

Risk factors for community-onset urinary tract infections caused by extended-spectrum ß-lactamase-producing Escherichia coli

Background/aim: Community-onset urinary tract infections(UTIs) caused by extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli have increased in many parts of the world. This study aimed to determine the prevalence and risk factors for community-onset UTI caused by ESBL-producing E. coli. Materials and methods: This prospective cohort study was conducted between January 2012 and March 2014 in cases of community-onset UTI caused by E. coli. Patients with UTI due to ESBL-producing E. coli and patients with UTI due to non-ESBL-producing E. coliwere compared to identify risk factors for ESBL-producing E. coli in the community. Results: A total of 305 patients (116 males [46.4%]; mean age: 57.76 ± 18.06 years) were included in the study. Among these patients, 154 (50.5%) were infected with ESBL-producing E. coli. In multivariate analysis, the healthcare-associated UTI (odds ratio [OR]: 1.80; 95% confidence interval [CI]: 1.02­3.18; P = 0.041), upper urinary tract infection (OR: 3.05; 95% CI: 1.76­5.29; P < 0.0001), use of antibiotics in the preceding 6 months (OR: 2.28; 95% CI: 1.21­4.30; P = 0.011), and having two or more risk factors (OR: 4.03; 95% CI: 1.73­9.35; P = 0.001) were the significant factors associated with increased risk of community-onset UTIs due to ESBL-producing E. coli. Conclusion: The increasing prevalence ofESBL-producing E. coli makes it difficult to decide the empirical therapy in UTIs, especially in patients with two or more of the risk factors. A better understanding of the epidemiology and risk factors associated with community-onset UTIs due to ESBL-producing E. coli may have significant implications in decision-making for empirical antimicrobial treatment.

[Prevalence and genotype distribution of human papillomavirus in patients attending to gynecology polyclinics]. / Jinekoloji poliklinigine basvuran hastalarda insan papillomavirüs prevalansi ve genotip dagilimi.

Human papillomavirus (HPV) infection is the most significant risk factor of the development of cervical cancer. The distribution of HPV prevalence and genotype varies widely between regions. In this study, it was aimed to investigate the prevalence and genotype distribution of HPV, retrospectively. One thousand one hundred and seventy patients who applied to the department of obstetrics and gynecology were included in this study. Samples were collected from patients for cervical HPV DNA and Pap smear. The Pap samples taken for Pap smear were fixed with alcohol and analyzed according to the modified Bethesda system. HPV identification and typing were performed using the "Linear Array HPV Genotyping Test (Roche Molecular System, USA)". Patients were divided into 5 groups due to their age. Total HPV ratio was most frequently found among the patients who were between 17-30 years old, while HR-HPV was most frequently found between 51-60 years. Nine hundred seventy-eight of 1170 (83.6%) patients had normal cytologic findings whereas 192 (16.4%) had abnormal cytologic findings. HPV was detected in 37.2% of the total patients. high-risk HPV (HR-HPV) rate was 21.2%, probable high risk (PR-HPV) rate was 6.4% and low risk HPV (LR-HPV) rate was 9.5%. When the relationship between cytologic findings and HPV was examined, normal cytology/HPV negative 67.8%; abnormal cytology/HPV negative 37.5%, normal cytology/HPV positive 32.2%, abnormal cytology/HPV positive 62.5% were detected. The highest prevalence of HPV was 8.9% with HPV 16, followed by 6, 53 and 52/53/35/58. A total of 354 patients were biopsied, 177 of whom were normal, 111 of whom were cervical intraepithelial neoplasia (CIN) 1, 66 of whom were CIN 2 and over. In the group with normal pathological findings, HR-HPV ratio was found as 15.8%, while in CIN 1 44.1% in CIN 2-3 63.6%. Sensitivity, specificity, positive predictive value, and negative predictive value of screening tests were examined in CIN 2 and more lesions. Sensitivity and specificity for HR-HPV were 63.6% and 73.3%, respectively, the same rates were 81.8% and 58.7% for HPV. The highest sensitivity was found in combination of HRHPV and Pap smear, the highest specificity in HPV. In conclusion, the HPV prevalence and genotype distribution in our study are similar to those reported in the world, but higher than previous studies in our country. These results may be due to our methodology and hospital based nature of our study group. We conclude that only smear or HR-HPV testing are not sufficient as a single pronged screening test, and that the participation of other genotypes of HPV in screening increases the sensitivity.

The Prevalence of Mixed Genotype Infections in Turkish Patients with Hepatitis C: a Multicentered Assessment.

BACKGROUND: HCV virus infections are one of the major health problems in the world that can cause cirrhosis and liver cancer at a higher rate than other hepatitis data. The aim of this study was to determine the prevalence of mixed infections with different HCV genotypes in Turkey and also to evaluate the current HCV genotype and sub-type distributions by a multicentered assessment. METHODS: The HCV genotype data of 17,578 hepatitis C patients collected from 23 centers from different geographic regions covering all Turkey were collected. The data included information about the HCV genotypes in the last 10 years (between 2007 and 2016), demographic properties of the patients and the methods/systems used to determine the genotypes. RESULTS: Two hundred twenty-eight of the patients (1.3%) had mixed genotype. The most common mixed genotype combination was 1b + 4 (0.83%) followed by 1a + 1b (0.26%). Genotype distribution varies according to geographical regions. However, genotype 1 (82.92%) was the most common genotype in all regions and all years. This was followed by genotype 3 (7.07%) and genotype 4 (5.43%). A variety of methods were used by the centers including sequencing, pyrosequencing, real-time PCR, in-house RFLP, reverse hybridization (LIPA), and hybridization. CONCLUSIONS: Infection with mixed HCV genotypes in Turkey is uncommon. Genotype distribution varies according to geographic regions; the most common genotype 1 is encountered all over the country, while genotypes 3 and 4 are only in some of the centers. Since there is limited information about mixed HCV infection, further investigations are needed to determine the clinical importance of mixed HCV infection.

[Evaluation of blood culture practices: Use of system (Epicenter) data]. / Kan kültürü uygulamalarinin degerlendirilmesi: Isletim sistemi (Epicenter) verilerinin kullanimi.

Sepsis is a serious clinical problem and estimated to be responsible for 18 million annual deaths worldwide. Therefore, the use and the rapid processing of blood cultures are important for the transition from empiric therapy to directed therapy. The aim of this study was to assess the best blood culture practices in Turkey. We have examined the collection practices and techniques at four different hospitals, and a total of 165.443 blood culture bottles were evaluated (2013-2015). At the preanalytical phase most of the data which were important and which could support hospital quality systems/practices were not entered into the HIS and EpiCenter system. At the analytical phase loading of the bottles and removal of positive bottles primarily occurred between 6:00 and 9:00 AM but the positivity rate of the bottles showed a homogeneous distribution throughout the day. In other words, there were significant delays at processing positive blood culture bottles related to laboratory workers. The effect of education regarding best practices, transition from single bottle to two bottle cultures was successful in all hospitals. Single bottle usage decreased below 10% in all hospitals. Significantly more positive cultures were detected at multiple cultures when compared with the single bottle collection practice. In retrospective patient records, it was seen that all the laboratories reported the results of Gram staining to the clinics. However, these data were not recorded to the EpiCenter. The contamination rates of Ankara Numune Hospital and Akdeniz University Faculty of Medicine Hospital are 6.2% and 5.4% respectively, contamination rates were not reported in other hospitals. The most common isolates detected in blood cultures were Escherichia coli, Klebsiella pneumoniae, Enterococcus faecium, Staphylococcus aureus, and Acinetobacter baumannii. The mean time for the detection of these organisms were less than 20 hours in the aerobic bottle and anaerobic bottles. A total of 79.6% of facultative anaerobic isolates were detected in both bottles; 9.8% were detected only in the aerobic bottles; 10.6% of the isolates were detected only in the anaerobic bottles. As a result, the educational efforts in Turkey have met with success for transition from collecting single bottle blood culture sets to two bottle blood cultures. However, further efforts are needed to increase the number of blood culture sets collected during a 24 hours' period. In addition, errors at the preanalytical, analytical and postanalytical periods (taking samples, loading bottles into the system and processing positive blood cultures) should be eliminated.

[Comparison of genotypic and phenotypic characteristics in biofilm production of Staphylococcus aureus isolates]. / Staphylococcus aureus izolatlarinda biyofilm üretiminin genotipik ve fenotipik karakterlerinin karsilastirilmasi.

Staphylococcus aureus which is an important pathogen, is known to have several virulence factors. The pathogenicity of S.aureus isolates are related with features like adherence, various toxins, enzymes, structural and extracellular factors. Slime factor and biofilm formation are also the pathogenicity factors of the bacteria. Biofilm formation is usually associated with chronic infections and has become a subject of interest in a wide area of research. Biofilm is an adherent structure formed by bacteria encased within a matrix produced on natural body surfaces or medical devices. As the biofilm producing S.aureus isolates are more resistant to antibiotics and and their biofilms prevent phagocytosis, the treatment of infections caused by biofilm positive S.aureus isolates become difficult. The icaADBC locus and some proteins have provento be responsible for the formation of S.aureus type biofilm. The aim of this study was to investigate the relationship between the genotypic and phenotypic detection methods of biofilm formation with icaA, icaD and bap genes and phenotype expressions which are responsible for the formation of biofilm in S.aureus isolates. One hundred seventy five S.aureus isolates from various clinical specimens were included in the study. For the phenotypic detection of biofilm producing isolates Congo red agar (CRA) medium and microplate method were used. The biofilm-producing strain Staphylococcus epidermidis ATCC 35984 and S. epidermidis ATCC 12228 were used as positive and negative controls, respectively. One hundred fifty two S.aureus biofilm producing isolates were detected at least by either Congo Red agar or microplate method and all isolates were screened for icaA, icaD and bap genes by PCR. The production of polysaccharide intracellular adhesins /poly-N-acetyl-beta-1-6-glucosamine (PIA/PNAG) was also investigated in S.aureus isolates. For the detection of PIA/PNAG chemiluminescence dot-blot method was used. According to the phenotypic evaluations based on colony morphologies in CRA medium, biofilm formation were found as negative in 101 of 175 isolates (57.7%), while biofilm formation were positive in 74 (42.3%) of the isolates. As a result of quantitative evaluation by microplate method based on absorbance measurement, biofilm production was determined as negative in 34 (19.4%) specimens, moderate in 112 (64.0%) specimens and strong in 29 (16.6%) specimens. Microplate and CRA methods were incompatible with each other when compared for their biofilm production determination (p< 0.001). Among the 152 clinical isolates used to determine the presence of icaA and icaD genes responsible for the formation of biofilms, the genes were detected in 136 (89.5%) of the isolates and in the S.epidermidis ATCC 35984 strain used as a positive control. icaA and icaD genes were not detected in sixteen of the 152 (10.5%) clinical isolates and in the S.epidermidis ATCC 12228 strain used as a negative control. A weak-moderate correlation was found between icaA and icaD genes and biofilm production determined in CRA medium. A good correlation was found between icaA and icaD genes and biofilm production detected in microplate method. bap gene was determined in only one of the 152 clinical S.aureus isolates studied and in S.aureus V329 strain used as positive control. For the detection of PIA/PNAG which was synthesized by icaADBC genes, 50 clinical S.aureus isolates were used. PNAG production was determined in 42 isolates with positive icaA and icaD genes by the chemiluminescence dot-blot method, and no PNAG production was determined among eight isolates with negative ica genes. As a result, it was found that the microplate method was more sensitive and reliable for the phenotypic determination of biofilm formation in S.aureus isolates, high level of biofilm formation among clinical S.aureus isolates (about 80%), the role of icaA and icaD genes and products (PIA/PNAG) in biofilm production was determined.

[Evaluation of virulence factors in enterococcus species]. / Enterokok türlerinin virulans faktörlerinin arastirilmasi.

Enterococci have recently become important due to their increased isolation rates in community-based and nosocomial infections and resistance to many antibiotics, including glycopeptides. The aim of this study was to evaluate the antimicrobial susceptible patterns and virulence factors of various clinical specimens; urine (n= 149), blood (n= 38), wound (n= 17), stool (n= 13), and other (n= 12) with a total of 229 enterococci including 138 E.faecalis and 91 E.faecium isolates. Aggregation factor (AF), enterococcus surface protein (esp), cytolysins and gelatinase encoding genes (asa1, esp, cylM, cylBcyl A, cylll, cylls, gelE, respectively) were investigated by molecular methods. Haemolysin production and gelatinase were studied phenotypically. A total of 30 isolates, 29 of E.faecium and one of E.faecalis isolates were resistant to vancomycin. High-level gentamicin and high-level streptomycin resistance in E.faecalis were 40.7% and 63.7% however, they were 47.1% and 55.8% in E.faecalis isolates. All strains were susceptible to linezolid. Ampicillin, penicillin and vancomycin resistance in E.faecium isolates were found to be higher than E.faecalis isolates (p= 0.001, p= 0.008 and p< 0.001). Asa1 (p< 0.001), cylll (p= 0.002) and cylls (p< 0.001) as well as gelatinase activity in isolates of E.faecalis were significantly higher than the isolates of E.faecium (p< 0.001). The most common virulence genes in our study were asa1 gene (45%), cyLs gene (33.2%) and esp gene (32.3%). Ciprofloxacin resistance in cylLL and cyLs gene positive isolates of E.faecalis were significantly higher compared to isolates that do not contain these genes (p= 0.035 and p= 0.047). Likewise, haemolysin producing E.faecium isolates were significantly more resistant to vancomycin compared to isolates that do not produce hemolysin (p< 0.001). When the virulence factors of vancomycin resistant and susceptible isolates were compared, the esp gene level in VRE E.faecium isolates was found to be 24.1%, while no esp gene was found in VRE E.faecalis isolates. The existence of asa1was negative in both VRE E.faecium and VRE E.faecalis isolates. The activity of hemolysin was found 42.3% for E.faecalis and 19.3% for E.faecium. In vancomycin-sensitive enterococcus (VSE) species, esp gene activity was 35.1% for E.faecalis, 29.4% for E.faecium, asa1 gene activity was 60.8% for E.faecalis and 47.1% for E.faecium, hemolysin activity was 52.8% for E.faecalis and 23.5% for E.faecium. In our study, it was found that VSE isolates have more virulence genes than VRE isolates. It should be kept in mind that VRE can causeinfections which are difficult-to-treat especially in hospitalized patients and VSE have significant virulence factors that can cause severe infections.

Fosfomycin Addition to Poly(D,L-Lactide) Coating Does Not Affect Prophylaxis Efficacy in Rat Implant-Related Infection Model, But That of Gentamicin Does.

Gentamicin is the preferred antimicrobial agent used in implant coating for the prevention of implant-related infections (IRI). However, the present heavy local and systemic administration of gentamicin can lead to increased resistance, which has made its future use uncertain, together with related preventive technologies. Fosfomycin is an alternative antimicrobial agent that lacks the cross-resistance presented by other classes of antibiotics. We evaluated the efficacy of prophylaxis of 10% fosfomycin-containing poly(D,L-lactide) (PDL) coated K-wires in a rat IRI model and compared it with uncoated (Control 1), PDL-coated (Control 2), and 10% gentamicin-containing PDL-coated groups with a single layer of coating. Stainless steel K-wires were implanted and methicillin-resistant Staphylococcus aureus (ATCC 43300) suspensions (103 CFU/10 µl) were injected into a cavity in the left tibiae. Thereafter, K-wires were removed and cultured in tryptic soy broth and then 5% sheep blood agar mediums. Sliced sections were removed from the tibiae, stained with hematoxylin-eosin, and semi-quantitatively evaluated with X-rays. The addition of fosfomycin into PDL did not affect the X-ray and histopathological evaluation scores; however, the addition of gentamicin lowered them. The addition of gentamicin showed a protective effect after the 28th day of X-ray evaluations. PDL-only coating provided no protection, while adding fosfomycin to PDL offered a 20% level protection and adding gentamicin offered 80%. Furthermore, there were 103 CFU level growths in the gentamicin-added group, while the other groups had 105. Thus, the addition of fosfomycin to PDL does not affect the efficacy of prophylaxis, but the addition of gentamicin does. We therefore do not advise the use of fosfomycin as a single antimicrobial agent in coating for IRI prophylaxis.

[Investigation of the effect of ibuprofen on wound healing in experimental Staphylococcus aureus soft tissue infections]. / Deneysel Staphylococcus aureus yumusak doku enfeksiyonlarinda ibuprofenin yara iyilesmesi üzerindeki etkisinin arastirilmasi.

Soft tissue infections (STIs) occur as a result of the colonization of pathogenic bacteria upon the destruction of normal skin microbial flora and the skin integrity. Streptococci and staphylococci are the most frequent causes of bacterial STIs. Non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen are often used in STIs because of their analgesic and antipyretic effects. However, evidence suggests that these drugs might delay both epithelization and angiogenesis in the early phases of wound healing because of an antiproliferative effect. The aim of this study was to investigate the effect of ibuprofen on the wound healing in STIs caused by Staphylococcus aureus in immunosuppressed mice. A total of 120 female Balb/c mice were used in the study and the mice were assigned to four test groups and two control groups. The test groups were defined as follows; B (Bacteria group, n= 23), BI (Bacteria + Ibuprofen group, n= 23), BA (Bacteria + Ampicillin group, n= 23), BIA (Bacteria + Ampicillin + Ibuprofen group, n= 21); and the control groups were defined as follows; S1B2 (only immunosuppressed controls, n= 15) and S2B2 (Sham group). Immunosupression was induced with cyclophosphamide and the experimental infection was generated by subcutaneous inoculation of bacterial suspension (2 x 10(8) cfu/ml) of methicillin-sensitive S.aureus ATCC 25923 to the right hind leg. Ibuprofen was given to the mice by gastric gavage (50 mg/kg/day), and ampicillin (100 mg/kg/day) by intramuscular injection. Wound sizes that appear in the animals were measured on a daily basis. Serum and tissue (epithelial tissue, connective tissue, sebaceous glands, sweat glands) samples were obtained on the first, third and seventh days. The tissue samples were examined histopathologically by hematoxylin-eosin (HE) staining method and IL-1, IL-6, TNF-α and VEGF (Vascular Endothelial Growth Factor) levels were determined in serum samples by ELISA method. The tissue cytokine reactions were also evaluated by immunohistochemical (immunoperoxidase staining) method in tissue samples. In our study, no significant change was detected in the wound sizes of B and BI groups from the second day to the end of study period (p> 0.05). On the other hand the wound dimensions of BA and BIA groups gradually decreased and remained superficial. The average serum levels of TNF-α and IL-1 was detected low in all groups. The mean value of serum IL-6 on the first day in group B was determined to be higher compared to other groups, and when this difference was compared to groups BI and BA, and the control group, it was found statistically significant (p< 0.05). In addition, the VEGF levels which were detected low in all groups in the third day of infection increased significantly at the seventh day. The results of histopathologic and immunohistochemical studies have supported the results of ELISA and yielded similar results with serum cytokine patterns. In conclusion, our data indicated that ibuprofen has no negative effect on the wound healing in soft-tissue infections caused by S.aureus.

[Investigation of macrolide, lincosamide and streptogramin B resistance in Staphylococcus aureus strains isolated from clinical samples by phenotypical and genotypical methods]. / Klinik örneklerden izole edilen Staphylococcus aureus suslarinda makrolid, linkozamid ve streptogramin B direncinin fenotipik ve genotipik yöntemlerle arastirilmasi

Staphylococcus aureus is one of the most common cause of both community and healthcare-associated infections. As staphylococci have developed resistance to various antibiotics, initially to penicillins then to methicillin and glycopeptides and have the ability to cause epidemics, they continue to be a major problem from past to present. Methicillin resistance gave rise to the use of alternative antibiotics such as macrolides, however worldwide development of macrolide resistance limited the use of these antibiotics. Macrolide resistance occurs either through target site modification (MLS(B) phenotype, encoded by erm genes), efflux pumps (MS phenotype, encoded by msrA/B genes) or decreased cell wall permeability. The aim of this study was to investigate the MLS(B) resistance of clinical S.aureus strains with phenotypic and genotypic methods. A total of 404 S.aureus strains isolated from different clinical samples (50% wound, 15% tracheal aspirate and 35% other samples) of inpatients (93.3%) and outpatients (6.7%) were included in the study. Double disc synergy test (D-test) was used for the phenotypical research and PCR was used for the genotypical research of MLS(B) resistance of isolates. One hundred fifty eight (39.1%) of the S.aureus isolates were methicillin-resistant (MRSA), and 246 (60.9%) were methicillin-susceptible (MSSA). By the use of D-test, constitutive (cMLS(B)) and inducible (iMLS(B)) clindamycin resistance were detected in 19 and 111 isolates, respectively, while five isolates were MS phenotype and 268 isolates were S phenotype (susceptible to erythromycin and clindamycin). The resistance genes of 136 isolates with MLS(B) resistance phenotype were determined genotypically and among 111 isolates showing iMLS(B) phenotype ermA gene was found in 81.9% (83 MRSA, 8 MSSA), ermC gene in 10.8% (7 MRSA, 5 MSSA), msrA gene in 10.8% (11 MRSA, 1 MSSA), msrB gene in 1.8% (2 MRSA) and ermB gene in 0.9% (1 MRSA). Among 19 strains with cMLS(B) phenotype, ermA was found in 57.9% (10 MRSA, 1 MSSA), ermC in 36.8% (6 MRSA, 1 MSSA) and ermB in 15.8% (3 MRSA). Among five strains with MS phenotype, ermA was found in 80% (2 MRSA, 2 MSSA), msrA in 75% (3 MSSA), msrB in 50% (2 MSSA) and ermC in 25% (1 MSSA) of the isolates. ErmA and ermC genes were detected together in 14 isolates, ermA, ermC and msrA genes in one isolate, ermA and msrA genes in 11 isolates, ermA, msrA and msrB genes in three isolates and ermA and ermB genes in three isolates, respectively. In this study, two MRSA isolates with MS phenotype and negative D-test had only ermA gene and among two MSSA strains, erm genes were also determined in addition to msr genes. In our study RAPD-PCR method was used to investigate the clonal similarity, however no dominance of one or a number of clonal type was observed among the isolates in which the resistance genes were identified. In conclusion, the detection of MLS(B) resistance in S.aureus isolates is likely to influence the selection of antibiotics in the treatment of the infections caused by this bacteria.

Seroprevalence of hepatitis e virus among primary school children.

OBJECTIVES: To investigate the seroprevalence of anti-hepatitis E virus antibody among primary school children in the two different areas of Denizli, Turkey. Methodology : Anti-HEV antibodies were investigated in 185 primary school children (91 from rural areas and 94 from urban areas of Denizli). The children were divided into two age groups as seven-year old group and fourteen-year old group. Samples were tested for anti-HEV Ab by an enzyme-linked immunoassay. Results : A total of 23 primary school children were anti-HEV Ab positive, giving a prevalence of 12.4%. The seroprevalence rate was 13.1% in rural areas and 11.7% in urban areas. The difference in the seropositive rates was not statistically significant (p>0.05). Among 185 primary school children, Anti-HEV antibodies were positive 17 (18.1%) in seven-year old group, and 6 (6.6%) in fourteen-year old group. The difference in the seropositive rates was statistically significant (p<0.05). Conclusions : There was no association between the anti-HEV Ab and gender, socioeconomic level, parental educational level, rural or urban areas. Anti-HEV Ab seroprevalence was higher in seven-year old children than fourteen-year old children.