In vitro activity of methylene blue and eosin methylene blue agar on colistin-resistant A. baumannii: an experimental study.

Introduction. Colistin is a last-resort antibiotic used against carbapenem-resistant Acinetobacter baumannii (AB); however, colistin resistance has been reported recently. Methylene blue (MB) is used in microbiology for staining, and in medicine as an antidote drug.Aim. We aimed to investigate the antimicrobial effects of MB and eosin methylene blue (EMB) agar against colistin-resistant AB strains.Methodology. Firstly, a standard strain and AB clinical isolate were included in the study. After determining MICs, two strains were transformed into colistin-resistant forms, using Li's method. At each step, new MICs were determined and subcultures were inoculated onto EMB and sheep blood agar (SBA). Colistin MICs of the subcultures were also determined using Mueller-Hinton agar (MHA) containing MB. Secondly, colistin-resistant clones from 31 multidrug-resistant AB clinical isolates were screened to investigate their susceptibilities to EMB agar.Results. In the first round, MICs of both strains had risen to 64 µg ml-1. Subpopulations with high colistin resistance were inhibited by MB and EMB agar, but could grow well on SBA. In MHA plates containing MB, the MICs decreased to 0.5 µg ml-1 for colistin-susceptible or moderately resistant clones. Additionally, clones with high colistin resistance showed atypical colony morphology on SBA. In the second round, 35 % of the clinical isolates, which had gained resistance to colistin, were inhibited by EMB agar.Conclusion. MB may have inhibitory effects against colistin-resistant AB. Secondly, using only EMB agar for subculturing may cause missing of colistin-resistant strains.

Antimicrobial susceptibility and resistance mechanisms of methicillin resistant Staphylococcus aureus isolated from 12 Hospitals in Turkey.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important nosocomial pathogens and is also emerging in Turkish hospitals. The aim of this study was to determine the antimicrobial susceptibility profiles of MRSA isolated from Turkish hospitals. MATERIALS AND METHODS: A total of 397 MRSA strains isolated from 12 hospitals in Turkey were included to present study. Antimicrobial susceptibilities were tested using agar dilution method. Presence of ermA, ermB, ermC, msrA, tetM, tetK, linA and aac-aph genes were studied by PCR. RESULTS: All strains were susceptible to vancomycin and linezolid. The susceptibility rates for fusidic acid, lincomycin, erythromycin, tetracyclin, gentamycin, kanamycin, and, ciprofloxacin were 91.9%, 41.1%, 27.2%, 11.8%, 8.5%, 8.3% and 6.8%, respectively. Lincomycin inactivation was positive for 3 isolates. Of 225 erythromycin resistant isolates 48 had ermA, 20 had ermC, and 128 had ermA-C. PCR was negative for 15 strains. Of 3 isolates with lincomycin inactivation one had linA and msrA. Of 358 gentamycin resistant isolates 334 had aac-aph and 24 were negatives. Among 350 tetracyclin resistant isolates 314 had tetM. Of 36 tetM negative isolates 10 had tetK. CONCLUSION: MRSA isolates from Turkish hospitals were multiresistant to antimicrobials. Quinolone and gentamycin resistance levels were high and macrolide and lincosamide resistance were relatively low. Susceptibility rates for fusidic asid were high. Linezolide and vancomycin resistance are not emerged. The most common resistance genes were ermA, tetM and aac-aph. Evolution of antimicrobial susceptibilities and resistance genes profiles of MRSA isolates should be surveyed at regional and national level for accurate treatment of patients and to control dissemination of resistance genes.

Conjunctivitis due to Neisseria sicca: a case report.

We report the first case, in Medline-based literature, of conjunctivitis caused by gram negative diplococcus, Neisseria sicca. Although it is not widely accepted as such, isolation from cultures of repeated eye swab samples suggests that N. sicca may be a pathogen in conjunctival infections. Positive culture for this organism should not be readily dismissed. Such conjunctivitis responded favorably to treatment with netilmicin eye drops.

[t030 is the most common spa type among methicillin-resistant Staphylococcus aureus strains isolated from Turkish hospitals]. / t030, Türkiye'deki Hastanelerden Izole Edilen Metisiline Dirençli Staphylococcus aureus Izolatlari Arasinda En Yaygin spa Tipidir.

Staphylococcus aureus is one of the most frequent agents causing hospital infections. S.aureus has a great ability to adapt itself to variety of conditions and successful clones can be epidemic and even pandemic by its ability spread from one continent to another. The aims of this study were to detect spa types of 397 methicillin-resistant S.aureus (MRSA) strains isolated from 12 centers in different geographical regions of Turkey from 2006 to 2008, and to investigate their clonality by PFGE and MLST typing. Additionally, 91 MRSA from four of those 12 centers isolated during 2011 were also studied for their spa types. PFGE profiles indicated the presence of a major pulsotype, namely pulsotype A with a rate of 91.4% (363/397), followed by pulsotype B (n= 18, 4.5%) and pulsotype C (n= 11, 2.8%). Among isolates tested 363 (91.4%) were SCCmec type III, 30 (7.6%) were SCCmec type IV. Sequence analysis of representative isolates revealed that ST239 (85.1%) was the most common MLST type followed by two MLST types ST737 (4%), and ST97 (2.8%), both SCCmec type IV. Two isolates were ST80 with SCCmec type IV. Of 397 isolates, 338 (85.1%) were t030, followed by t005 (2.5%) and t632 (2%). Among MRSA isolated during 2011, 64 (70.3%) of 91 were t030, 4 (4.4%) were t005, 2 (2.2%) were t015, and 2 (2.2%) were t1094. Among centers the t030 prevalence of 2006-2008 isolates ranged from 59-100%. The highest t030 prevalence was found in Ankara (100%) and lowest in Trabzon (59%) provinces which are located at central and northestern Anatolia, respectively. In Istanbul province, the prevalence of t030 was 94.5% among 2006-2008 isolates which decreased to 55.5% among 2011 isolates. Also a decrease in t030 rates was observed among samples from Konya and Trabzon but not from Aydin. Our results showed that the most common MRSA clone in Turkey is ST 239-SCCmec type III, t030 which persisted during the six years of the study period. Presence of PVL toxin gene was tested by PCR and 5 (3%) isolates found to be positive, of them two were SCCmec Type IV-ST80 and three were SCCmec Type III-ST239. This study is the largest epidemiological survey ever done in Turkey which showed presence of a hospital Turkish clone TR09 (ST239-SCCmecIII-t030) and a community clone TR10 (ST737-SCCmecIV-t005) largely disseminated in Turkey.

[Epidemiological evaluation of a rapidly-prevented tularemia outbreak in Canakkale province, Turkey]. / Çanakkale'de Hizla Önlenen Bir Tularemi Salgininin Epidemiyolojik Olarak Degerlendirilmesi.

Tularemia is a disease caused by Francisella tularensis and widely seen at northern hemisphere of the world. In Turkey, oropharyngeal infections caused by a less virulent serotype F.tularensis subsp. Holarctica are more prevalent. The aim of this study was to present the results of an epidemiological research performed after the detection of tularemia cases from Biga county of Canakkale province, Turkey, in December 2009. Following the report of two tularemia suspected cases from two villages (Baliklicesme and Sinekci) of Biga, an epidemiological investigation was undertaken to inspect the situation in this area. Water samples, clinical samples as throat swabs, wound swabs and serum samples were collected. Samples were cultured on heart agar supplemented with sheep blood, cysteine and antibiotics. Cultures were incubated at 37°C in 5% CO(2) and followed for 10 days. Suspected colonies were identified by slide agglutination test using F.tularensis antisera. F.tularensis antibodies were investigated by standard tube agglutination method. Positive results obtained with agglutination test were also checked for a probable crossreaction with Brucella antibodies by Rose-Bengal test. Water and wound samples were investigated using real-time polymerase chain reaction (RT Taqman PCR; Quantica, Techne Inc, UK) with probe and primers specific for ISFtu2 gene. All of the cultures yielded negative results, however eight of 16 water samples, one lymph node aspirate and one throat sample were found positive in F.tularensis TaqMan RT-PCR test. In tube agglutination test positive antibody titers between 1:20-1:1280 were detected in 36 of 115 serum samples. Two cases with antibody titers of 1:1280 and accompanying acute clinical findings, were diagnosed as tularemia and treated accordingly. Lymphatic drainage fluid samples obtained from one of these patients yielded positive result in PCR, however clinical sample could not be obtained from the other patient. The only epidemiological linkage between these acute cases (n= 2) and the other seropositive subjects (n= 34) was the use of local water supply system. It was learned that water obtained through reverse osmosis system had been used as drinking water at Baliklicesme village. Pre- and post-reverse osmosis system water samples from Baliklicesme village and samples from water supply of Sinekci village revealed positive results for F.tularensis by PCR. Since the only epidemiological relation between these two villages was using local water supply, tularemia cases encountered in this area were attributed to a waterborne epidemic and an automatic chlorination system was set up at each water reservoir in these villages. The establishment of these preventive measures curbed the growth of the epidemic. The cases presenting with throat sore, fever, lymphadenopathy (more than 2 cm), non-responsive to beta-lactam antibiotics, should be further investigated for tularemia. This work emphasizes that systematic setup and control of water disinfection systems are crucial to prevent tularemia outbreaks. Community and related authorities should be educated about the importance of water sanitation and chlorination.

Indoor and outdoor airborne bacteria in child day-care centers in Edirne City (Turkey), seasonal distribution and influence of meteorological factors.

This paper presents information about airborne mesophilic bacteria in the indoor and outdoor air of child day-care centers (CDCCs) in the city of Edirne, Turkey. Air samples were collected using the Petri plate gravitational settling method from the indoor and outdoor air of CDCCs. Counts of airborne bacteria were measured as colony forming units (CFU) collected by gravity onto Brain Heart Infusion Agar plates (with 5% sheep blood). Samples were taken monthly over a period of 12 months between January and December 2004. A total of 3,120 bacteria colonies were counted on 192 Petri plates. Four groups of culturable bacteria were identified: gram-positive cocci, gram-positive bacilli, endospore-forming gram-positive bacilli, and gram-negative bacteria. Airborne gram-positive bacteria were the most abundant at more than 95% of the measured population. While gram-positive cocci were more common in indoor environments, gram-positive bacilli were more dominant in outdoor air. Bacteria commonly isolated from CDCCs were identified at a genus level. Staphylococcus (39.16%), Bacillus (18.46%), Corynebacterium (16.25%), and Micrococcus (7.21%) were dominant among the genera identified in the present study. The dominant genera identified in the day-care centers were Staphylococcus, Micrococcus, and Corynebacterium for indoor air and Bacillus, Corynebacterium, and Staphylococcus for outdoor air. Staphylococcus, Streptococcus, Bacillus, and Corynebacterium genera were found in samples from every month. Bacterial colony counts were compared by sampling location (indoors and outdoors), seasons, and meteorological factors. We found negative correlations between the monthly total outdoor bacterial counts and the sampling day's average relative humidity and average rainfall, and the monthly average rainfall. Fluctuations in bacterial counts in different seasons were observed.

Tularemia re-emerging in European part of Turkey after 60 years.

The aim of this study was to investigate a tularemia outbreak in the Thrace region of Turkey. The outbreak occurred in Demirkoy village of Edirne, in 2005. Of 400 villagers, 266 were examined and their sera were taken. Throat swabs and lymph node aspirates were cultured. Specific antibodies in patients and domestic animals were screened by a microagglutination test. PCR assays and cultures of the samples of patients, animal tissues, and water sources were performed, along with active surveillance to identify risk factors. Seven out of 10 cases were diagnosed as oropharyngeal form; the remaining three patients were asymptomatic. The cultures for tularemia were negative; however, PCR assays were positive in one lymph node aspirate and in water from one spring. Some animals had the specific antibody at low levels. Increased rodent population in the vicinity, exposure to wild rabbits, and drinking from one of the springs were identified as risk factors with the risk ratios (and 95% confidence interval) of 10.5 (10.3-10.7), 6.5 (5.43-7.57), and 2.1 (1.1-2.5), respectively. Therapeutic and preventive measures were taken. When tularemia cases have been detected in a region even a few decades earlier, tularemia should be considered in the differential diagnosis of patients.

Nocardial brain abscess: review of clinical management.

Nocardiosis has become a significant opportunistic infection over the last two decades as the number of immunocompromised individuals has grown worldwide. We present two patients with nocardial brain abscess. The first patient was a 39-year-old woman with systemic lupus erythematosus. A left temporoparietal abscess was detected and aspirated through a burr-hole. Nocardia farcinica infection was diagnosed. The patient had an accompanying pulmonary infection and was thus treated with imipenem and amikacine for 3 weeks. She received oral minocycline for 1 year. The second patient was a 43-year-old man who was being treated with corticosteroids for glomerulonephritis. He was diagnosed with a ring-enhancing multiloculated abscess in the left cerebellar hemisphere, with an additional two small supratentorial lesions and triventricular hydrocephalus. Gross total excision of the cerebellar abscess was performed via a left suboccipital craniectomy. Culture revealed Nocardia asteroides, and the patient was successfully treated with intravenous ceftriaxone, then oral trimethoprime-sulfamethoxazole for 1 year. The clinical course, radiological findings, and management of nocardial brain abscess are discussed in light of the relevant literature, and current clinical management is reviewed through examination of the cases presented here.

First case report of empyema caused by Beauveria bassiana.

Infections with Beauveria bassiana are extremely rare in humans. A 51-year-old man was admitted to hospital with the diagnosis of lung adenocarcinoma. Thoracic wall resection with lobectomy was applied and empyema has developed after prolonged air leakage. B. bassiana was isolated in pleural fluid. The patient improved without antifungal therapy after thoracotomy with securing of air leakage.

Rubella seroprevalence among Turkish adolescent girls living in Edirne, Turkey.

This study was designed to estimate the rubella seroprevalence in unvaccinated Turkish adolescent girls in urban and rural areas of Edirne, and to create preventive strategies for congenital rubella syndrome (CRS). The sample, representing 12- to 17-year-old adolescent girls, consisted of 1,600 subjects selected from school lists by systematic and random sampling, which was matched by age and urban-rural residency strata proportional to the corresponding distributions in the Edirne population. For each participant, a questionnaire was completed and rubella-specific IgG antibodies were measured. After analysis of samples, seropositivity prevalence, equivocal and seronegative samples of adolescent girls in Edirne were determined as 93.1%, 0.6% and 6.3%, respectively. Data from the present study may indicate that 6.9% of adolescent girls have considerable risk for rubella infection during pregnancy. Eliminating rubella and CRS in Turkey will require national health service efforts, including vaccination of all adolescents and all susceptible women of childbearing age.

The antimicrobial susceptibility of Stenotrophomonas maltophilia isolates using three different methods and their genetic relatedness.

BACKGROUND: Stenotrophomonas maltophilia is inherently resistant to many antimicrobials. So far, antimicrobial susceptibility tests for S. maltophilia have not been fully standardized. The purpose of the study was to compare the susceptibility of S. maltophilia isolates against seven different antimicrobials using three different methods and to investigate their genetic relatedness. RESULTS: Although trimethoprim/sulfamethoxazole (SXT) and ciprofloxacin have the lowest MIC values, SXT (98.1%) and ticarcillin/clavulanate (TLc) (73.1%) were found to be the most effective antimicrobials by agar dilution method, which was in accordance with the breakpoints established by NCCLS. Disc diffusion and E-test was in agreement with agar dilution method for SXT. When the isolation dates, clinics, antibiotyping, and AP-PCR data were investigated, two small outbreaks consisting of five and three cases were determined. CONCLUSION: By using the NCCLS criteria, disc diffusion and E-test were unreliable alternative methods for S. maltophilia, except for SXT. However, the significance of these data should be confirmed by further experimental and clinical studies.

[Salmonella enterica serotypes and Salmonella infections: a multicenter study covering ten provinces in Turkey]. / Salmonella enterica serotipleri ve Salmonella enfeksiyonlari: Türkiye'de on ili kapsayan çok merkezli bir çalisma.

In order to find the distinctive features of Salmonellae and Salmonella infections in Turkey, 620 Salmonellae strains, isolated from various clinical samples (481 stool, 108 blood, 12 urine, 3 bone marrow, 3 cerebrospinal fluid, 9 pus, and one from each of the bile, pleural fluid, wound, catheter samples) in 13 clinical microbiology laboratories of 10 provinces in Turkey (Ankara, Antalya, Bursa, Edirne, Eskisehir, Istanbul, Izmir, Kayseri, Konya and Trabzon) between July 1, 2000 and June 30, 2002, were serotyped. Among the patients 43% were female, 57% were male, 63.2% were from outpatient clinics and 36.8% were hospitalized patients. Seventy eight percent of the patients had gastroenteritis, 10.7% had septicemia/local infection, 9.8% had typhoid/paratyphoid fever and 1.5% were carriers. Incidence of gastroenteritis was higher in 0-5 years age group (p<0.001). Of the 620 Salmonella enterica isolates, 47.7% were S. Enteritidis, 34.7% S. Typhimurium, 6% S. Paratyphi B, 2.9% S. Typhi, 0.2% S. paratyphi A, 6.1% serogroup C1, and 2.4% serogroup C2. S. Enteritidis was the most common serotype in all provinces except for Kayseri, where S. Typhimurium was found to be the most common serotype (68.2%). Overall, the most frequently isolated serotype was S. Enteritidis, also being the most common serotype in stool and blood cultures. During the surveillance period two outbreaks have occurred, the first one by S. Enteritidis strains in Edirne, and the second one by S. Typhimurium strains in Kayseri. As a result, Salmonella infections are still a common health problem in Turkey, and active surveillance of Salmonella infections has vital importance.

[Characteristics of malaria cases diagnosed in Edirne province between 1994-2002]. / Edirne'de 1994-2002 yillari arasinda saptanan sitma olgularinin özellikleri.

In this study, the epidemiological characteristics of malaria cases in Edirne province were investigated. Between the years of 1994-2002, a total of 317,087 blood samples were collected from soldiers in the province with selective active surveillance and from the resident population with active or passive surveillance methods, by the medical staff of Malaria Control Department and Health Centers, to search the presence of Plasmodium. In 281 of them Plasmodium spp. were detected, and the characteristics of malaria cases were investigated. Of the cases, 238 (84.7%) were detected in the first three years and mostly in September. While the indigenous cases were detected in the districts where rice planted intensely, the imported cases were detected in the districts heavily populated by military staff. Of the imported cases, 62% originated from Diyarbakir, Batman and Sanliurfa provinces (Southeast part of Turkey). P. vivax was detected as the causative agent in all blood samples except one P. ovale. This latter case has been the only one in Turkey so far and he was a student from Afghanistan. Attaching importance to fight off mosquitoes in intensely rice planted districts and strictly surveying the military staff, particularly from the region of Southern-East Anatolia, have led to successful control of the malaria cases in Edirne region.

The epidemiology of hepatitis a virus infection in children, in Edirne, Turkey.

Turkey is a middle endemic area with respect to Hepatitis A virus (HAV) infection. However, the frequency of this infection varies due to socio-economic differences in various regions. The aim of this study was to detect the most likely age of exposure to HAV and factors affecting infection rates among children living in Edirne. A sample of 645 children between the ages of 0-19 living in Edirne were tested for total anti-HAV levels using ELISA method. A questionnaire on socio-economic status (SES), possible risk factors, and place of residence was completed for each child. Anti-HAV seropositivity was found to be 4.4, 25, 37.3 and 43.2%, in 2-5, 6-10, 11-14 and 15-19 age groups, respectively. Seropositivity was found to be increasing with age (p < 0.05). According to logistic regression analysis results; mother's education, SES of family, history of hepatitis in primary family members and the number of brothers or sisters were determined as factors increasing the seropositivity of HAV. Furthermore, HAV infection risk was found to be a decreasing function of income that is higher the income less likely the infection. These results showed that HAV infection rate in Edirne is in middle endemicity and the most likely way of exposure is transmission from family members. Although it requires further cost-effectiveness studies, our results indicate that applying the HAV vaccination in early childhood would be beneficial to decrease the prevalence of the infection and prevent HAV epidemics.

Resistance to macrolides in Group A streptococci from the European section of Turkey: genetic and phenotypic characterization.

The aim of this study was to determine the susceptibilities to macrolides of Group A streptococcal isolates from the European section of Turkey. In the case of resistant isolates, the patterns and genetic mechanisms of erythromycin resistance were studied. Seven (2.7%) of the 260 isolates were resistant to erythromycin. Four of them showed the M phenotype and harboured mefA genes whereas three isolates showed the inducible macrolide, lincosamide and streptogramin B resistance phenotype and harboured ermTR genes. In the European section of Turkey, the current resistance rate of Group A streptococci to macrolides remains low.

Clinical importance of extended-spectrum beta-lactamase (PER-1-type)-producing Acinetobacter spp. and Pseudomonas aeruginosa strains.

Recently, an extended-spectrum beta-lactamase (PER-1) was found to be disseminated among Acinetobacter spp. and Pseudomonasaeruginosa isolates in Turkey. A population-based cohort study was conducted to elucidate predictive mortality factors in patients with nosocomial infections caused by Acinetobacter spp. and P. aeruginosa, with particular reference to PER-1-type extended-spectrum beta-lactamase (ESBL) production. The study group comprised 16 and 21 non-survivors and 82 and 126 survivors in cohorts infected with Acinetobacter and P. aeruginosa, respectively. In the Acinetobacter-infected cohort, nosocomial pneumonia, hypotension and infection with a PER-positive isolate were independent predictors of mortality. In the P. aeruginosa-infected cohort, impaired consciousness, a PER-positive isolate, male sex and (with a negative relative risk) urinary tract infection were independent predictors of death. This study demonstrated the relationship of PER-1-type ESBL-producing Acinetobacter spp. and P. aeruginosa with poor clinical outcome.