Tularemia seroprevalence in humans in the region of the Hittite-Arzawa War (Inner Aegean Region), where the first biological weapon was used 3300 years ago.

BACKGROUND: : According to Egyptian records, tularemia emerged in the Canaan region, where it was first identified and spread to Anatolia over the Euphrates. It was used as an active biological weapon for the first time in the Hittite-Arzawa War in 1320-1318 BC. This study aimed to investigate the seroprevalence of tularemia in the Inner Aegean Region, which is thought to be the region where this war was fought 3300 years ago. METHODS: Tularemia seropositivity in humans was investigated in 27 villages/neighborhoods in 3 districts in each of Manisa, Kütahya, and Usak provinces. Before the study, the participants were informed about the disease via posters, and their blood samples were taken following filling out the questionnaire. Microagglutination tests were performed using in-house tularemia antigen and V plate for serological experiments. Rose-Bengal test was also performed on seropositive sera. RESULTS: Of the total of 410 people, 226 (55.12%) were male. The mean age of the volunteers was 43.72 years. The highest participation was from Kütahya Province. According to the results of the tularemia microagglutination test, seropositivity was detected in 6 cases. It was determined that all of the seropositive volunteers were in Kütahya. When the tularemia antibody titers were examined, seropositivity was determined at 1/20-1/160 titers. No positivity was detected in the Rose-Bengal test for cross-reaction. DISCUSSION: Kütahya has been identified as a risky region in terms of tularemia in the Inner Aegean Region. In order to use the resources in the country economically, first of all, the risk areas in terms of tularemia should be determined by serological studies in all regions. In order to increase awareness about the disease, physicians and filiation teams should be trained in risky areas. Surveillance studies should be conducted to identify and monitor possible sources in areas identified as risky.

Frequency of azole resistance in clinical and environmental strains of Aspergillus fumigatus in Turkey: a multicentre study.

OBJECTIVES: Aspergillus fumigatus causes several diseases in humans and azole resistance in A. fumigatus strains is an important issue. The aim of this multicentre epidemiological study was to investigate the prevalence of azole resistance in clinical and environmental A. fumigatus isolates in Turkey. METHODS: Twenty-one centres participated in this study from 1 May 2018 to 1 October 2019. One participant from each centre was asked to collect environmental and clinical A. fumigatus isolates. Azole resistance was screened for using EUCAST agar screening methodology (EUCAST E.DEF 10.1) and was confirmed by the EUCAST E.DEF 9.3 reference microdilution method. Isolates with a phenotypic resistance pattern were sequenced for the cyp51A gene and microsatellite genotyping was used to determine the genetic relationships between the resistant strains. RESULTS: In total, resistance was found in 1.3% of the strains that were isolated from environmental samples and 3.3% of the strains that were isolated from clinical samples. Mutations in the cyp51A gene were detected in 9 (47.4%) of the 19 azole-resistant isolates, all of which were found to be TR34/L98H mutations. Microsatellite genotyping clearly differentiated the strains with the TR34/L98H mutation in the cyp51A gene from the strains with no mutation in this gene. CONCLUSIONS: The rate of observed azole resistance of A. fumigatus isolates was low in this study, but the fact that more than half of the examined strains had the wild-type cyp51A gene supports the idea that other mechanisms of resistance are gradually increasing.

Recent Increase in the Prevalence of Fluconazole-Non-susceptible Candida tropicalis Blood Isolates in Turkey: Clinical Implication of Azole-Non-susceptible and Fluconazole Tolerant Phenotypes and Genotyping.

Candida tropicalis is the fourth leading cause of candidemia in Turkey. Although C. tropicalis isolates from 1997 to 2017 were characterized as fully susceptible to antifungals, the increasing global prevalence of azole-non-susceptible (ANS) C. tropicalis and the association between high fluconazole tolerance (HFT) and fluconazole therapeutic failure (FTF) prompted us to re-evaluate azole susceptibility of C. tropicalis in Turkey. In this study, 161 C. tropicalis blood isolates from seven clinical centers were identified by ITS rDNA sequencing, genotyped by multilocus microsatellite typing, and tested for susceptibility to five azoles, two echinocandins, and amphotericin B (AMB); antifungal resistance mechanisms were assessed by sequencing of ERG11 and FKS1 genes. The results indicated that C. tropicalis isolates, which belonged to 125 genotypes grouped into 11 clusters, were fully susceptible to echinocandins and AMB; however, 18.6% of them had the ANS phenotype but only two carried the ANS-conferring mutation (Y132F). HFT was recorded in 52 isolates, 10 of which were also ANS. Large proportions of patients infected with ANS and HFT isolates (89 and 40.7%, respectively) showed FTF. Patients infected with azole-susceptible or ANS isolates did not differ in mortality, which, however, was significantly lower for those infected with HFT isolates (P = 0.007). There were significant differences in mortality (P = 0.02), ANS (P = 0.012), and HFT (P = 0.007) among genotype clusters. The alarming increase in the prevalence of C. tropicalis blood isolates with ANS and HFT in Turkey and the notable FTF rate should be a matter of public health concern.

Combined Effectiveness of Honey and Immunonutrition on Bacterial Translocation Secondary to Obstructive Jaundice in Rats: Experimental Study.

BACKGROUND Obstructive jaundice is a serious, life-threatening condition that can lead to death as a result of sepsis and multiorgan failure due to bacterial translocation. Treatment should be started as soon as possible after diagnosis. MATERIAL AND METHODS Forty 24-week-old male Sprague Dawley rats, with an average weight of 250 g to 300 g, were included in this study. The rats were randomly placed into five groups, each group consisted of eight rats. The sham group underwent only common bile duct (CBD) dissection and no ligation was performed. CBD ligation was applied to the other groups. After the operation, one CBD group was fed with rat chow only, the others were fed with rat chow supplemented with honey, or immunonutrients, or honey plus immunonutrients. After 10 to 12 days, all rats were sacrificed; blood and tissue samples were collected for biochemical, microbiological, and histopathological evaluation. RESULTS In the groups that were fed with honey and immunonutrients, alanine aminotransferase (ALT) levels were decreased significantly compared to the other groups. Statistically significant differences were detected in terms of bacterial translocation (BT) rates among liver and spleen samples, and laboratory values of serum, except for MLNs of the BDL+HI group, when compared to other groups. We found mean mucosal thickness of ileum samples have been improved notably in the BDL+HI group compared to the other groups, especially compared to the C/BDL group. CONCLUSIONS Immunonutrition applied with honey had immunostimulant effects, decreased BT due to an additive effect, and had positive effects on intestinal mucosa.

[Results of a multicenter study investigating plasmid mediated colistin resistance genes (mcr-1 and mcr-2) in clinical Enterobacteriaceae isolates from Turkey]. / Ülkemizde klinik Enterobacteriaceae izolatlarinda plazmit aracili kolistin direnç genlerini (mcr-1 ve mcr-2) arastiran çok merkezli çalismaya ait sonuçlar.

Colistin is a polymyxin antibiotic which is considered as one of the last line agents against infections due to multidrug resistant or carbapenem resistant gram-negative pathogens. Colistin resistance is associated with chromosomal alterations which can usually cause mutations in genes coding specific two component regulator systems. The first plasmid-mediated colistin resistance gene, mcr-1 was described in Escherichia coli and Klebsiella pneumoniae isolates in December 2015 and followed by another plasmid-mediated colistin resistance gene mcr-2 in 2016. The rapid and interspecies dissemination of plasmid-mediated resistance mechanisms through horizontal gene transfer, have made these genes considerably threatening. After the first reports, although mcr-1/mcr-2 producing Enterobacteriaceae isolates have been reported from many countries, there have been no reports from Turkey. Thus, the aim of this study was to investigate the presence of mcr-1/mcr-2 in clinical Enterobacteriaceae isolates from different parts of our country. A total of 329 Enterobacteriaceae isolates from 22 laboratories were collected which were isolated between March, 2015 and February, 2016. mcr-1/mcr-2 were investigated by polymerase chain reaction during February-March, 2016. Two hundred and seventeen of Klebsiella pneumoniae (66%), 75 of Salmonella spp. (22.8%), 31 of Esherichia coli (9.4%), 3 of Enterobacter cloacae (0.9%), 2 of Klebsiella oxytoca (0.6%) and 1 of Enterobacter aerogenes (0.3%) isolates were included to the study. Agarose gel electrophoresis results of PCR studies have shown expected band sizes for positive control isolates as 309 bp for mcr-1 and 567 bp for mcr-2. However, the presence of mcr-1/mcr-2 genes was not detected among the tested study isolates of Enterobacteriaceae. Although mcr-1/mcr-2 were not detected in our study isolates, it is highly important to understand the mechanism of resistance dissemination and determine the resistant isolates by considering that colistin is a last-line antibiotic against infections of multidrug or carbapenem resistant gram-negative bacteria. Thus, it is suggested that these mechanisms should be followed-up in both clinical and non-clinical (e.g. isolates from food animals, raw meats and environment) isolates of special populations.

[A rarely isolated bacterium in microbiology laboratories: Streptococcus uberis]. / Mikrobiyoloji laboratuvarlarinda nadir izole edilen bir etken: Streptococcus uberis.

Streptococcus uberis is a gram-positive bacterium that is mostly responsible for mastitis in cattle. The bacterium rarely has been associated with human infections. Conventional phenotyphic methods can be inadequate for the identification of S.uberis; and in microbiology laboratories S.uberis is confused with the other streptococci and enterococci isolates. Recently, molecular methods are recommended for the accurate identification of S.uberis isolates. The aim of this report is to present a lower respiratory tract infection case caused by S.uberis and the microbiological methods for identification of this bacterium. A 66-year-old male patient with squamous cell lung cancer who received radiotherapy was admitted in our hospital for the control. According to the chest X-Ray, patient was hospitalized with the prediagnosis of ''cavitary tumor, pulmonary abscess''. In the first day of the hospitalization, blood and sputum cultures were drawn. Blood culture was negative, however, Candida albicans was isolated in the sputum culture and it was estimated to be due to oral lesions. After two weeks from the hospitalization, sputum sample was taken from the patient since he had abnormal respiratory sounds and cough complaint. In the Gram stained smear of the sputum there were abundant leucocytes and gram-positive cocci, and S.uberis was isolated in both 5% sheep blood and chocolate agar media. Bacterial identification and antibiotic susceptibility tests were performed by VITEK 2 (Biomerieux, France) and also, the bacterium was identified by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) based VITEK MS system as S.uberis. The isolate was determined susceptible to ampicillin, erythromycin, clindamycin, levofloxacin, linezolid, penicillin, cefotaxime, ceftriaxone, tetracycline and vancomycin. 16S, 23S ribosomal RNA and 16S-23S intergenic spacer gene regions were amplified with specific primers and partial DNA sequence analysis of 16S rRNA polymerase chain reaction (PCR) products were performed by 3500xL Genetic Analyzer (Applied Biosystems, USA). According to the partial 16S rRNA gene sequencing results, bacterium was confirmed as S.uberis. This report makes a significant contribution to the number of case reports of human infections caused by S.uberis as the identification was performed by current microbiological methods in our case. In conclusion, S.uberis should be evaluated as an opportunistic pathogen among the immunosuppressed patients and in addition to phenotypic bacteriological methods, the other recent microbiological methods should also be utilized for the identification.

[Microbiological approach to a possible infective endocarditis case caused by Aggregatibacter actinomycetemcomitans]. / Aggregatibacter actinomycetemcomitans kaynakli olasi enfektif endokardit olgusuna yaklasim.

Aggregatibacter (Actinobacillus) actinomycetemcomitans, a small, gram-negative coccobacillus that grows slow and fastidious, is generally colonized in the oral cavity. It is a rarely seen bacterium because of the difficulty of isolation but it can be a causative agent for dental infections and infective endocarditis (IE) particularly in the persons having prosthetic heart valves. In this report, a possible IE case caused by A.actinomycetemcomitans in a patient with aortic valve replacement has been presented. A 36-year-old man has admitted to Trakya University Hospital, Health Center for Medical Research and Practice, with the complaints of chills, malaise, intermittent fever, severe arthralgia and weight loss (20 kg). During his follow-up period, the blood cultures that were obtained three week intervals yielded the identical gram-negative coccobacilli morphology. The patient was then diagnosed as possible IE on the basis of having one major (growth of the typical microorganisms that may cause IE in two different blood cultures) and two minor (presence of prosthetic valve and high fever) criterias. The isolate could not be identified with conventional methods, while it was identified as Francisella tularensis with VITEK 2 (bioMerieux, France) system. Hence this identification was not confirmed by real-time Taqman polymerase chain reaction, so MALDI-TOF mass spectrometry was used to identify this bacteria. In the first run of the study, the isolate was named as Shigella dysenteriae initially, however when it was retested the next day it was identified as A.actinomycetemcomitans. In order to enlighten these conflicting results, 16S and 23S ribosomal DNA sequence analysis was performed, and consequently the bacterium was identified as A.actinomycetemcomitans. Doxycycline (2 x 100 mg po, 20 days) and streptomycin (2 x 10 mg/kg im, 10 days) therapy were initiated, considering the initial suspicious identification (F.tularensis), and on the fifth day of therapy the blood culture was negative with the regression of patient's complaints. Therapy continued with the addition of rifampicin to doxycycline from the 21(st) day and the patient discharged with cure. As a result, the identification of an exceptional bacterium like A.actinomycetemcomitans may be difficult and time-consuming in certain laboratory facilities. So, the use of different identification methods in addition to classical methods are needed to overcome such a problem, especially for uncommon isolates and clinically discordant cases. This case was presented because A.actinomycetemcomitans is a rare etiological agent for IE patients and it could be a good example to draw attention to the problem when identifying the organism using automatized identification systems.

Epidemiology of tularemia.

Tularemia is considered to have existed in Anatolia for several thousand years. There are suspicions regarding its use in biological warfare in the Neshite-Arzawan conflict. The causative agent of tularemia may have first been used as a biological weapon in 1320-1318 BC. The disease has recently become a significant re-emerging disease globally as well as in Turkey. In the period of 2001-2010, Kosovo had the highest annual incidence in Europe at a rate of 5.2 per 100,000. Sweden, Finland, Slovakia, Czech Republic, Norway, Serbia-Montenegro, Hungary, Bulgaria, and Croatia follow with rates of 2.80, 1.19, 1.0, 0.81, 0.42, 0.4, 0.36, 0.21, and 0.15 per 100,000 people, respectively. Tularemia in Turkey was first reported in the soldiers living in the region very close to the Kaynarca Stream of Thrace in 1936. It has started to gain more and more importance, especially in recent decades in Turkey, due to a very high number of cases and its spread throughout the country. A total of 431 tularemia cases were recorded in Turkey in 2005, but a significant reduction was observed in the number of the cases in the next three years; the number of patients decreased to 71 in 2008. The number of cases increased again in 2009 and continued in subsequent years. The number of cases reached 428, 1531, 2151, and 607 in 2009, 2010, 2011, and 2012, respectively. The number of cases peaked in 2011 in Turkey, and was in fact higher than the total number of cases in all European Union countries. The number of cases is higher in females than males in Turkey. In Turkey, 52% of cases of tularemia diagnoses occur from December to March and the most common clinical presentation is the oropharyngeal form caused by contaminated water. Rodents are the most likely sources of tularemia outbreaks in Turkey as well as in Kosovo. Organisms such as ticks, flies and mosquitoes are vectors of tularemia transmission to mammals. Because ticks can carry the bacteria by both transovarial and transstadial transmission, they play a role in the life cycle of tularemia as both reservoir and vector.

[Investigation of the presence of Francisella tularensis by culture, serology and molecular methods in mice of Thrace Region, Turkey]. / Trakya Bölgesi'nde farelerde kültür, seroloji ve moleküler yöntemlerle Francisella tularensis varliginin aranmasi

Tularemia is a disease that has been reported in Turkey since 1936. Although mice are considered to have a role in the transmission of Francisella tularensis to man, this has not been exactly confirmed yet. The aim of this study was to investigate the presence of F. tularensis in mice by using culture, serology and molecular methods. For this purpose, four villages (Edirne-Demirkoy, Kirklareli-Kaynarca, Tekirdag-Muzruplu, Tekirdag-Sinanli) were selected in Thrace Region of Turkey where tularemia cases had been reported previously. A total of 126 live-catch mouse traps were established in warehouses, barns, areas near wells, water tanks and creeks in the villages in December 2012. Traps were kept overnight and the next day the animals collected were identified at species-level. The live-captured mice were anesthetized and their heart blood samples were obtained. Subsequently, liver and spleen tissues were removed from every mouse under aseptic conditions in the class-2 safety cabinet. These tissues were cultivated in Francis medium containing 5% sheep blood, 0.1% cystein, 1% glucose and incubated for seven days in both normal atmosphere and 5% carbondioxide incubator at 37°C. Tularemia microagglutination test was performed by using the sera which were obtained from live-captured mice. Finally, DNAs were isolated from both liver and spleen tissues of mice, and real-time polymerase chain reaction (Tularemia RT-PCR; Public Health Agency of Turkey, Ankara) were performed. In our study, a total of 19 mice were captured and of these 11 were alive. Ten mice were identified as Apodemus flavicollis, seven were Mus macedonicus and two were Mus musculus. There were no Francisella tularensis isolation in the cultures of mice liver and spleen tissues. Serological tests yielded negative results for 10 mice whose serum samples could be obtained. In RT-PCR, positivity were detected in spleen tissues of two mice which were captured from Kaynarca where first tularemia cases in Turkey in 1936 were reported but has no report from then on. One of them was a live female Mus macedonicus, and the other was a dead male Apodemus flavicollis. In quantitative evaluation, number of microorganism per organ were calculated as 4 x 103 cfu/spleen in Mus macedonicus and 4 x 104 cfu/spleen in Apodemus flavicollis. This is the first study in Turkey indicating that the mice in natural environment harbored F.tularensis. In conclusion, the results of this study indicated that the agent of tularemia has been retained since 1936 in Kaynarca region and this persistence might present a potential risk for tularemia epidemics.

Combined effects of tauroursodeoxycholic Acid and glutamine on bacterial translocation in obstructive jaundiced rats:.

BACKGROUND: Bacterial Translocation is believed to be an important factor on mortality and morbidity in Obstructive Jaundiced. AIMS: We investigated the probable or estimated positive effects of tauroursodeoxycholic acid, which has antibacterial and regulatory effects on intestinal flora, together with glutamine on BT in an experimental obstructive jaundiced rat model. STUDY DESIGN: Animal experimentation. METHODS: Forty adult, male, Sprague Dawley rats were used in this study. Animals were randomised and divided into five groups of eight each: sham (Sh); control (common bile duct ligation, CBDL); and supplementation groups administered tauroursodeoxycholic acid (CBDL+T), glutamine (CBDL+G), or tauroursodeoxycholic acid plus glutamine (CBDL+TG). Blood and liver, spleen, MLN, and ileal samples were taken via laparotomy under sterile conditions for investigation of bacterial translocation and intestinal mucosal integrity and hepatic function tests on the tenth postoperative day. RESULTS: There were statistically significant differences in BT rates in all samples except the spleen of the CBDL+TG group compared with the CBDL group (p=0.041, p=0.026, and p=0.041, respectively). CONCLUSION: It is essential to protect hepatic functions besides maintaining intestinal mucosal integrity in the active struggle against BT occurring in obstructive jaundice. The positive effect on intestinal mucosal integrity can be increased if glutamine is used with tauroursodeoxycholic acid, which also has hepatoprotective and immunomodulatory features.

A Case Report on Aspergillus lentulus Pneumonia.

BACKGROUND: Aspergillus lentulus was described as a new species in 2005 but it was isolated from Turkey for the first time. CASE REPORT: A. lentulus was isolated as the cause of pneumonia from a patient who had renal transplantation 4 months ago. The patient received immunosuppressive treatment after transplantation. A. lentulus was isolated from his sputum as an agent in pneumonia developed 4 months after the transplantation. Leukocytes, blastospores, and hyphae were seen in both Gram- and Giemsastained smears of the sputum. The isolate was identified by using the Maren A. Klich algorithm and molecular methods and confirmed by the reference laboratory of the CBS Fungal Biodiversity Centre (The Netherlands). In the susceptibility tests of the isolate, minimal inhibitory concentrations for amphotericin B, voriconazole, posaconazole, and caspofungin were found to be 0.5 µg/mL, 0.25 µg/mL, 0.125 µg/mL, and 0.25 µg/mL, respectively. The patient recovered with voriconazole treatment (2×200 mg/day). CONCLUSION: The use of the molecular tests is important for identification of A. lentulus strains because they are very easily confused with A.fumigatus strains according to phenotypic characteristics.

[Antifungal susceptibilities of Aspergillus spp. strains isolated from invasive aspergillosis cases]. / Invazif aspergillozlu olgulardan izole edilen Aspergillus spp. suslarinin antifungal duyarliliklari.

Aspergillus species found abundantly in the outer environment and hospital setting may lead to serious morbidity and mortality particularly in patients with suppressed immunity. This retrospective study was aimed to investigate the antifungal susceptibilities of Aspergillus spp. isolated from aspergillosis cases being hospitalized. Aspergillus spp. isolated from samples of the patients with suspected fungal infections between January of 2002 and October of 2007, were investigated. A total of 678 samples (420 lower respiratory tract, 202 sterile body fluids, and 56 biopsy/tissue specimens) from 569 patients were included in the study. The samples were incubated in 25 degrees C and 35 degrees C on brain-heart-infusion agar supplemented with blood and on Sabouraud dextrose agar. Gram and Giemsa stained samples were also examined by microscopy. Mold type of fungi were identified by conventional techniques. "Invasive aspergillosis" was described according to criteria of Invasive Fungal Infections Cooperative Group of the European Organization for Research and Treatment of Cancer. A. fumigatus (n = 8), A. flavus (n = 2) and A. niger (n = 2) were isolated from 12 patients' samples (2.1%), 9 of them were lower respiratory tract and one of each was ascid, brain biopsy and pleural fluid specimens. All of those patients have had an underlying diseases such as malignancy. The susceptibility of the isolates to caspofungin, voriconazole, itraconazole and amphotericin B was tested by broth microdilution susceptibility testing and to posaconazole by E-test (AB Biodisk, Sweden). The lowest minimum inhibitory concentration (MIC) (< or = 0.125 microg/ml) values were detected for caspofungin and posaconazole for Aspergillus spp., however, the highest MIC values were detected for amphotericin B (> 1 microg/ml). MIC values of the all strains except one, were detected as < or = 0.5 microg/ml for voriconazole and itraconazole. In one A. niger strain itraconazole MIC value was 2 microg/ml. Since the number of other species was low, MIC50 value was determined only for A. fumigatus strains and it was found that the highest MIC50 value was for amphotericin B (2 microg/ml) and the lowest MIC50 values were for posaconazole (0.064 microg/ml), caspofungin (0.064 microg/ml), itraconazol (0.25 microg/ml) and voriconazol (0.25 microg/ml). Since caspofungin and posaconazole revealed the lowest MIC values, they should be taken into consideration in choice of therapy of aspergillosis cases in our hospital.

[Cutaneous infection caused by Alternaria alternata in an immunocompetent host]. / Immün sistemi saglam bir konakta Alternaria alternata ile olusan deri enfeksiyonu.

Human infections caused by Alternaria alternata are more frequently reported in immunosupressive hosts. In this report, a rarely seen cutaneous infection, caused by A. alternata in an immunocompetent patient was presented. The patient (71-years-old, male) was admitted to the dermatology unit with complaints of an erythematous and squamatous lesion of 5 cm diameter on left malleolar region. The case was evaluated as immunocompetent based on the normal serum total immunoglobulin and complement levels, anti-HIV negativity, and no known underlying disease. A number of Alternaria spp. conidia and hypha were seen in the microscopical examination of KOH treated cutaneous scrapings of the lesion obtained in two different days. Fungal cultures of the skin scrapings yielded the growth of a fungus identified as A. alternata. Although fungal elements were not detected in haematoxylene-eosin stained smears of the skin biopsy, A. alternata was again isolated in the culture of the biopsy specimen. The identification of the fungus was confirmed by a reference center (Mycology Section of Scientific Institute of Public Health, Belgium) and it was integrated to BCCM/IHEM collection under accession number IHEM 22598. Antifungal susceptibility test efforts failed due to a problem in the preparation of fungal suspension. Oral itraconazole (200 mg/day) and bifonazole cream was used for the treatment and the lesion regressed after the 19th day of the therapy. The treatment was continued with oral and local terbinafine for two weeks and the patient fully recovered. Since A. alternata was demonstrated both in the skin scrapings and tissue biopsy through microscopic examination and culture, it was evaluated as the causative agent of skin infection rather than colonization. This was the first A. alternata infection in an immunocompetent patient in the light of the current literature.

Characteristics of the Turkish isolates of Francisella tularensis.

In this study, cultures of patients with tularemia were evaluated, and antimicrobial susceptibilities of two Francisella tularensis strains were tested by disk diffusion and E-test methods. A high-resolution multiple-locus variable-number tandem repeat analysis (MLVA) comprising six variable-number tandem repeat loci was applied to elucidate the genetic relatedness among Turkish and Bulgarian isolates which were isolated in a recent outbreak. The patients were diagnosed in two outbreaks in two cities of Turkey in 2005 and 2006. A total of 16 samples from 12 patients were cultured, and PCR tests were carried out on 15 samples that were positive in five lymph node aspirates and two soft tissue aspirates. F. tularensis was isolated from the lymph nodes of two patients. Aminoglycosides, quinolones, chloramphenicole, tetracyclines, nitrofurantoin, and rifampicin inhibited growth of the isolates. The Turkish isolates appeared to share a common MLVA pattern with one of the four Bulgarian outbreak genotypes.

[Investigation of the agents and risk factors of dermatophytosis: a hospital-based study]. / Dermatofitozlarda etkenlerin ve risk faktörlerinin arastirilmasi: hastane bazli bir Calisma.

The aims of this study were the detection of distribution of dermatophyte species isolated from the clinical samples of patients with dermatophytosis and the evaluation of risk factors for the development of dermatophytosis. A total of 441 skin, nail and scalp/hair specimens obtained from 301 patients (151 were male; age range 2 months-80 years, median 42 years) and 884 foot and hand skin and nail specimens obtained from 221 control subjects (110 were male; age range 5-75 years, median 36 years) were included to the study between the period of January to December 2005. All the samples have been evaluated by direct microscopic (DM) examination and by culture. A total of 121 (40.2%) patients yielded positivity for dermatophytes, of them 63 were positive by both DM and culture methods, seven were only culture positive, and 51 were only DM positive. Nine (9.8%) of 92 culture positive samples from 70 patients were found negative in DM, while 85 (50.6%) of 168 DM positive samples from 114 patients were negative in culture. 23.5% (12/51) of DM positive but culture negative patients were given antifungal therapy previously. The most prominent species isolated from the cultures were Trichophyton rubrum with a rate of 68.4% (63/92), followed by T. mentagrophytes (18.4%); T. violaceum (3.3%); T. verrucosum, T. tonsurans and Epidermophyton floccosum (2.2% for each); T. schoenleini, Microsporum canis and Trichophyton sp. (1.1% for each). Of the patient samples whose cultures were positive, 45% were from the foot skin. The presence rate of dermatophytes in controls was found as 3.2% (7/221); T. rubrum was isolated from the foot skin of five and T. mentagrophytes was isolated in toenail of two control subjects. About 42% of the samples belonged to the patients who admitted to hospital between December to February period. The evaluation of the risk factors revealed that presence of trauma, pet contact, ritual cleansing and diabetes mellitus had no effect on the development of dermatophytoses, however the presence of fungal infection in the family, male gender, some professions (being farmer, worker and retired), and the use of immunosupressive drugs have been found to increase the risk of dermatophytosis. The number of cases with dermatophytoses started to increase beginning from the age of 20 and peaked in the ages between 40-59 years old. As a result T. rubrum was determined as the most frequently isolated dermatophyte and tinea pedis was the most frequently observed clinical form in our hospital, emphasizing the importance of early diagnosis and effective treatment in superficial fungal infections which have high morbidity.

[Investigation of tularemia seroprevalence in the rural area of Thrace region in Turkey]. / Trakya bölgesinin köylerinde tularemi seroprevalansinin arastirilmasi.

The first published tularemia epidemic in Turkey had been reported in 1936 from Luleburgaz (located in European part-Thrace region-of Turkey), and the second was in 1945 again in the same province. Following a long period of time without any tularemia report from Thrace region, in 2005 another epidemic occurred in a village of Edirne, another province located in the same region. Since there is presumptive evidence of circulation of the infectious agent, Francisella tularensis in Thrace region of Turkey, a large scale seroepidemiological study is needed. In this study, the presence of antibodies against F. tularensis in 1782 subjects, choosen by "thirty cluster" method, inhabiting in 90 different villages of Edirne, Kirklareli, and Tekirdag provinces in Thrace Region, were investigated. The subjects were included to the study on the basis of volunteering (74.3% were male; mean age: 46 years; age range: 6-92 years) and demographical characteristics and their possible risky behaviours were recorded in a questionnaire form. Antibodies specific for F. tularensis were screened by microagglutination test, and were found positive in five (0.3%) of the subjects between the titers of 1/20- 1/160. All of the seropositive subjects were adult males (ages between 22-74 years); three were living in the two villages of Kirklareli, while the others were from the villages of Tekirdag and Edirne. Rose Bengal test was also found positive in three of the seropositive subjects, and with the thought of a probable cross reaction they were taken into an advanced investigation for brucellosis. The risk evaluation revealed that male gender, being together with livestock and exposure to ticks were the major risk factors. Since the data of this study indicated that F. tularensis is in circulation in Thrace Region, the educational programmes for both the healthcare workers and inhabitants of this region should be attempted for the prevention of a possible epidemic.

[Aspergillus spp. isolations from respiratory tract samples in Trakya University Hospital]. / Trakya Universitesi Hastanesi'nde solunum yolu örneklerinde Aspergillus spp. izolasyonlari.

The characteristics of cases diagnosed as aspergillosis and Aspergillus spp. strains isolated from the respiratory tract samples in Mycology Laboratory of Trakya University Hospital between January 2002 and May 2006 were investigated. In this period, 137 bronchoalveolar lavages, 95 sputum, nine tracheal aspirates, three lung biopsies and one bronchial biopsy of 85 patients were processed. The samples were incubated in 25 degrees C and 35 degrees C media by culturing on brain heart infusion agar with blood and Sabouraud dextrose agar. Presence of leucocytes and fungal structures were searched in the smear stained by Gram and Giemsa. The patient was defined as probable aspergillosis case, if he/she patient had clinical findings, lung infiltration or fungus ball radiologically, at least one risk factor predisposing to aspergillosis and isolation of Aspergillus spp. in lower respiratory tract samples without finding of other nonmycotic infection. Of 22 patients isolated Aspergillus spp., 13, six, two, one were internalized in chest diseases, haemotology, neurosurgery and oncology clinics, respectively. Seven positive cultures were considered as findings of aspergillosis. Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger were isolated in three, two, and two patients, respectively. Fungal structures were detected in only one sample in the direct microscopical examination. Ages of seven patients, five were males and two were females, were between 15 and 60. Predisposing risk factors were acute leukemia in six patients and lung cancer in one patient. Five patients were neutropenic and one was neutrophylic. Fungus ball was detected in radiological imaging of one patient, had a pulmonary cavitary lesion. Conventional amphotericine B was used in their therapies. Antifungal agents were switched to caspofungin and itraconazole in two and one patients, respectively. Three patients died in four weeks after isolation of Aspergillus spp. Aspergillosis cases were not high in our hospital because of absence of transplantation center for bone marrow or solid organ.

The effects of methylene blue on renal scarring due to pyelonephritis in rats.

The aim of this study was to evaluate the efficiency of methylene blue (MB) in preventing renal scar formation after the induction of pyelonephritis (PNP) in a rat model with delayed antimicrobial therapy. An inoculum of the K-12 strain of Escherichia coli was injected into both kidneys. Control groups received isotonic saline instead of bacterial solution. Four equal groups were then formed: the PNP group was untreated and the PNP ciprofloxacin (CIP) treated group was treated only with CIP intraperitoneally (i.p.) starting on the third day following bacterial inoculation. In the PNP (MB)-treated group, MB was given i.p., and in the PNP MB + CIP-treated group, MB + CIP were administered i.p.. In the sixth week following bacterial inoculation, all rats were sacrificed, and both kidneys of the rats in all groups were examined biochemically and histopathologically for renal scarring. Renal scar was significant in the groups treated with MB alone or MB + CIP combination compared with untreated or antibiotic only groups. Delayed treatment with antibiotics had no effect on scarring. These results suggest that the addition of MB to the delayed antibiotic therapy might be beneficial in preventing PNP-induced oxidative renal tissue damage.