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Introduction: The assessment of the clonal spread of methicillin resistant Staphylococcus aureus (MRSA) in nosocomial and communityacquired infections through characterization of the isolates is critical for tracking the evolution of the epidemics, implementing effective control measures, and preventing future outbreaks of MRSA. In this context, it is aimed with this study to determine the clonal relationships between the S. aureus isolates obtained from the patients receiving treatment in the intensive-care units of a state hospital in Turkey. Methodology: A total of 80 MRSA isolates obtained from the patients receiving treatment in three different intensive-care units were analyzed for their antibiotic susceptibilities, pulsed-field gel electrophoresis profiles, and multilocus sequence types. Results: The dendrogram of the pulsed-field gel electrophoresis profiles revealed two major pulsed-field gel electrophoresis pulsotypes: A and B, which were further divided into two (A1 and A2) and four (B1-B4) subgroups, respectively. Multilocus sequence typeanalysis indicated that all isolates belonged to a single MRSA clone, sequence type 239. No significant difference was found between the antibiotic sensitivity profiles of strains isolated from different intensive-care units. All of the strains were sensitive to linezolid and vancomycin. Conclusions: It was concluded that the MRSA strains isolated from the patients receiving treatment at the intensive-care units of the hospital constituted two major pulse-field types and belonged to the ST239 lineage, one of the most extensively distributed MRSA lineages throughout the world. [ABSTRACT FROM AUTHOR]

Salmonella spp. is the most frequently isolated foodborne pathogens causing human and animal diseases. The aim of this study was to investigate antimicrobial susceptibility profiles and the molecular typing of 200 Salmonella Enteritidis strains isolated from the patients' stools between 2016 and 2019 in Turkey. The isolates were examined for antibiotic susceptibility patterns (21 antimicrobial agents) by Kirby-Bauer disc diffusion method or molecular typing by Pulsed-field gel electrophoresis (PFGE) and plasmid profiling. Although all isolates were susceptible to four antibiotics (suphamethoxazole/trimethoprim, chloramphenicol, streptomycin and trimethoprim), all were resistant to 15 different antibiotics. In the PFGE study performed with XbaI enzyme, all isolates were found to be related to each other according to similarity rates of 85% and above. There were two major clones, clone A and B. Clone A was divided into 6 pulsotypes (A1-A2-A3-A4-A5-A6) and clone B was divided into 3 pulsotypes (B1-B2-B3). Clone A had 87% similarity and Clone B had 90% similarity. The clustering rate was 86% (172/200). All isolates harboured 1-4 plasmid ranging in size from 2.5 to 57 kb and showed 6 plasmid profiles (P1-P6). All isolates carried the 57 kb plasmid individually or in combination with other plasmids. Most of the isolates 136 (68%) had P2 profile. Our findings indicate that the majority of all isolates were clonally related and had cross contamination problems. In this study, the importance of molecular typing methods in order to take more effective protection and control measures against Salmonella has been demonstrated and proposed to use such methods. [ABSTRACT FROM AUTHOR]

A novel coronavirus (SARS-CoV-2) outbreak, responsible for a pneumonia-associated respiratory disorder (COVID-19), has started in early December 2019 in Wuhan, China, and has rapidly spread around the world. Rapid and accurate diagnostic testing plays a crucial role in tackling the COVID-19 pandemic. In this study, it was aimed to compare 5 primer sets designed to amplify different regions for the detection of SARS-CoV-2 and to perform sequence analysis. Conventional RT-PCR was carried out using primers targeting different regions of the virus genome including ORF1ab, Envelope (E), RNA-dependent RNA polymerase (RdRp), Spike (S) and Nucleocapsid (N) genes for the diagnosis of COVID-19. DNA sequence of ORF1ab gene from each sample were compared with the DNA sequence data of SARS-CoV-2 stored in the GenBank and ORF1ab phylogenetic tree was constructed. The amplicon sizes of ORF1ab, S, E, N and RdRp genes were 588 bp, 440 bp, 145 bp, 323 bp and 196 bp, respectively. The SARS-CoV-2 RNA was detected from 74% of total samples from RdRp gene, 87% for N gene, 74% for S gene, 61% for E gene and 82% for ORF1ab region. The ORF1ab sequences of SARS-CoV-2 from 82 patients were had 100% identity to the sequence of Wuhan isolate and among themselves. The phylogenetic analysis revealed that all isolates formed a cluster. The results of this study suggest that the N region is the best for SARS-CoV-2 identification. [ABSTRACT FROM AUTHOR]

The purpose of this study was to determine the phenotypic antibiotic susceptibility patterns, extended-spectrum β- lactamase (ESBL) genes and genotypic profiles of ESBL-positive Escherichia coli strains isolated from urine samples obtained from outpatients with urinary tract infection in Turkey. A total of 120 E. coli strains during 2017, 2018, and 2019 (40 patients per year) were examined for antibiotic susceptibility patterns by disc diffusion method, for ESBL genes using PCR and sequencing and for molecular typing by pulsed-field gel electrophoresis (PFGE) method. The isolates were evaluated for their sensitivity to 21 different antibiotics. Four different antimicrobial resistance patterns were determined according to antibiotic susceptibility status of the isolates. The β- lactamase genes detected in the isolates were CTX-M-15 + OXA-1 (n= 14), CTX-M-15 (n= 24), TEM-1 + CTX-M-15 (n= 52), TEM- 1 + SHV-12 (n=6), SHV-12 1 (n= 6), TEM-1 + CTX-M-1 (n= 6), TEM-1 + CTX-M-16 (n= 6) and TEM-1 + CTX-M-9 (n= 6). The CTX-M-15 was the most prevalent ESBL enzyme in the isolates. As a result of PFGE analysis performed by XbaI enzyme restriction process, one major PFGE profile and three main groups (Group I-II-III) were observed. While antibiotic resistance profiles of the strains showed four groups (RI-RII-RIII-RIV), PFGE band profiles showed a major group (90% similarity ratio). High ESBL production and decreased susceptibility to broad-spectrum cephalosporins were observed in E. coli strains. In addition, PFGE analysis showed high clonal similarity among E. coli isolates. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the prevalence and clonal typing of extended-spectrum beta-lactamase (ESBL)- producing Escherichia coli in healthy broilers in Turkey. Three hundred broiler cloacal samples were collected from various broiler slaughterhouses and inoculated on Levine agar plates supplemented with 2 μg/mL cefotaxime. Suspected strains were identified using a BBL Crystal Enteric/Nonfermenter ID Kit (Becton Dickinson, USA) and ESBL production was confirmed using an ESBL phenotypic confirmatory test. ESBL types were analyzed using PCR and sequencing. Pulsed field gel electrophoresis (PFGE) was performed with XbaI for the clonal typing of ESBL-producing E. coli isolates. While 33 phenotypic ESBL-producing E. coli isolates were identified, eight of them had only the blaTEM-1. Twenty-five ESBL-producing isolates were detected. This research is the first on the investigation and detection of ESBL-producing E. coli isolates from broilers in Turkey. In this study, 8.3% ESBL-producing E. coli were isolated from the cloacal samples of broilers collected from slaughterhouses in Turkey. CTX-M-15 (80%) was the most frequently isolated ESBL type. Using PFGE analysis, it was determined that these isolates had clonal similarity. [ABSTRACT FROM AUTHOR]

Background/aim: The purpose of the study was to evaluate the performance of the VITEK mass spectrometry (MS) (bioMérieux, France) system for the identification of Klebsiella spp. isolated from different sources. Moreover, while assessing the ability of the VITEK 2 automated expert system (AES) to recognize antimicrobial resistance patterns, the researchers have extended the study to compare VITEK 2 with the routine antimicrobial susceptibility testing method. Materials and methods: This study tested 51 Klebsiella spp. isolates that were isolated from environmental examples and clinical examples. Results of conventional methods and the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS were compared. Then, any differing results were compared against a reference 16S rRNA gene sequence, and when indicated, a recA sequencing analysis was done. Results: VITEK MS correctly identified 100% of the Klebsiella spp. isolates. There were two K. oxytoca isolates incorrectly identified to the species level with conventional methods according to the 16S rRNA gene sequencing analysis. In addition, a VITEK 2 AST-N261 card was used for the detection of extended spectrum beta-lactamases (ESBL). Using the VITEK 2 AES, ESBL positivity was found at the rate of 16.3% whereas this rate was 4.08% using the disk diffusion method. Conclusion: MALDI-TOF MS is a rapid and accurate method for the identification of Klebsiella spp. Moreover, the bioMérieux AES provides a useful laboratory tool for the interpretation of susceptibility results. [ABSTRACT FROM AUTHOR]

The species within the genus Bartonella are intracellular bacteria causing long-lasting bacteremia in humans and animals. In this study, Bartonella spp. in 173 small mammals, which were Apodemus flavicollis, A. witherbyi, A. uralensis, A. mystacinus, Myodes glareolus, Crocidura suaveolens, Rattus rattus and Rattus norvegicus species captured from Western Black Sea Region of Turkey, were investigated by blood culture and molecular methods. The positivity of Bartonella was 63.6% (110/173) by blood culture of small mammalian. The gltA gene regions for the isolated strains were identified by DNA sequencing analysis. Isolates were identified as Bartonella taylorii, B. birtlesii, B. coopersplainsensis and a zoonotic B. grahamii. [ABSTRACT FROM AUTHOR]

Bacillus cereus is an aerobic and facultatively anaerobic, spore-forming bacterium, and it is found naturally in soil and poses a risk factor for the contamination of food and foodstuffs including cereals, vegetables, spices, ready-to-eat (RTE) foods, meats, milk, and dairy products. This study determined the prevalence of B. cereus in raw poultry meat, raw cow's milk, cheese, spices, and RTE foods in Hatay province. The study also analysed the psychrotrophic properties, toxigenic characteristics, and pulsed-field gel electrophoresis (PFGE) profiles of the isolates. The levels of contamination with B. cereus determined for cheese, raw milk, RTE foods, spices, and raw poultry meat were 16.6%, 34.2%, 42.8%, 49%, and 55.5%, respectively. B. cereus was isolated from 84 (42%) of the 200 samples analysed and the 84 isolates were verified by PCR analysis targeting the haemolysin gene specific for B. cereus. Of the total isolates, 64 (76.1%) were psychrotrophic. The toxin gene profiling of B. cereus isolates was determined by amplifying the four genes nhe, hbl, cytK, and ces. The nhe and cytK genes were most frequently detected in the isolates, while the hbl and ces genes were not found. In addition, a high genetic relationship between the isolates was detected at a 92% similarity level by PFGE analysis. In conclusion, the occurrence of both psychrotrophic and toxigenic B. cereus strains in this study indicated a potential risk for food spoilage and food poisoning. [ABSTRACT FROM AUTHOR]

Francisella tularensis DNA extractions and isolates from the environment and humans were genetically characterized to elucidate environmental sources that cause human tularemia in Turkey. Extensive genetic diversity consistent with genotypes from human outbreaks was identified in environmental samples and confirmed water as a source of human tularemia in Turkey. [ABSTRACT FROM AUTHOR]

Q fever is a widespread zoonotic disease that is caused by obligate intracellular bacteria, Coxiella burnetii. This study was planned to determine the prevalence of C. burnetii in cows', goats', and ewes' bulk milk (BM) samples using PCR and to confirm positive results by DNA sequencing. A total of 150 BM samples (50 samples of each cows', goats', and ewes' milk) collected from 15 randomly selected dairy farms in Hatay province were analyzed. The BM samples were taken between January 2012 and July 2013. Bacterial DNA was extracted directly from milk samples. Nine of the BM samples (6 %) were PCR positive; five from cows' BM, two from ewes' BM, and two from goats' BM. Bacterial DNA was detected in 3 of 15 (20 %) dairy farms. Positive results were confirmed by DNA sequencing. The sequencing results of Coxiella DNA extracted from cows', goats', and ewes' milk samples were consistent with the reference strain with 100 %, 99.7 %, 99.8 % homology, respectively. BM samples were found to be contaminated with C. burnetii; therefore, taking hygienic measures is necessary for food safety and public health. [ABSTRACT FROM AUTHOR]

Mortality due to K. pneumoniae bacteremia is on rise, particularly in regions with high rates of carbapenem and colistin resistance. We aimed to define risk factors for colistin resistance and its impact on mortality. Patients diagnosed with "carbapenem-resistant K. pneumoniae (CRKp)" bacteremia between 2014 and 2018 were divided into two groups as "colistin susceptible (ColS)" and "colistin resistant (ColR)" based on broth microdilution method. Retrospective case-control study was conducted to compare characteristics and outcomes. Multiple logistic regression model was used to define independent risk factors for acquired colistin resistance and Cox proportional hazard model for 28-day mortality. A total of 82 patients (39 ColS and 43 ColR) were included. Mean age was 61.5 years, and 50 (61%) were male. Colistin resistance was significantly increased with duration of hospital stay (p = 0.007) and prior colistin use (p = 0.007). Overall, the 28-day mortality rate was 66%. Age (p = 0.014) and colistin resistance significantly increased 28-day (p = 0.009) mortality. Microbiological response to treatment within 7 days favors survival. PFGE analysis revealed an outbreak with K. pneumoniae ST78 and ST45 clones. Patients treated with combined antimicrobials had significantly lower 28-day mortality (p = 0.045) in comparison to monotherapy. However, types of combinations did not show significant superiority on each other. Colistin resistance increases 28-day mortality in CRKp bacteremia. Although combined regimens are more effective than monotherapy, existing antibacterial combinations have no apparent superiority to each other. New treatment options are pivotal. [ABSTRACT FROM AUTHOR]

Introduction: Stenotrophomonas maltophilia, which is able to form a biofilm, has mostly been related to catheters when it is the agent in hospital infections; these infections generally present as bacteremia and pneumonia, which may progress with complications and result in death. Methodology: The study included 153 S. maltophilia strains isolated from clinical samples sent to our hospital laboratory between 1 January 2014 and 30 June 2018. The bacteria were identified and their antibiotic sensitivity was determined using the VITEK-2 automated system. PFGE (Pulsed Field Gel Electrophoresis): The strains isolated from 34 patient clinical samples and from 1 patient bedcover were taken for PFGE examination. Results: The TMP/SXT and levofloxacin sensitivity of 153 S. maltophilia strains was examined. TMP/SXT resistance was determined to be 39% and levofloxacin resistance at 5%. Among 35 S. maltophilia strains, seven genotypes were identified using the PFGE method. While three strains showed a specific genotype profile, the other 32 were determined to consist of four clusters. The cluster rate was therefore 91.4% (32/35). Conclusions: There was a clonal relationship between the vast majority of the 35 S. maltophilia isolates, which suggests that there was a crosscontamination problem in the hospital. One strain (#4) was identified by dendrogram analysis showed a high rate of similarity to the other strains and was determined to be the common source of the cross-contamination. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to investigate the prevalence of the rotavirus and adenovirus related to the viral etiology of gastroenteritis as well as to determine the frequency of common rotavirus genotypes. Material and Method: Fecal samples were obtained from 492 children under 6 years of age who had not been vaccinated with the rotavirus vaccine and who applied to the pediatric outpatient clinic of Afyon Kocatepe University Hospital with the complaint of diarrhea. All of the fecal samples were evaluated in order to detennine the presence of bacterial and parasitic agents using conventional identification methods, all results were negative. Rotavirus and adenovirus antigens were examined in stool samples using the immunochromatographic method. Rotavirus G and P genotypes were determined by reverse transcription PCR using consensus primers detecting the VP7 and VP4 genes, followed by semi-nested type-specific multiplex PCR. Results: It was found that 3.3%, and 20.3% of 492 children with acute gastroenteritis were positive for adenovirus and rotavirus, respectively. A total of six different combinations of G and P types were found, including combinations of the Gl, G2, G4, G9, P[4], and P[8] genotypes. The most common rotavirus genotypes were G9P[8] (48.7%), followed by G9P[4] (17.5%). Other strains were G1P[8] (16.2%), G2P[8] (11.2%), G1P[4] (3.7%), and G4P[8] (2.5%). Sixty-six percent of the regional rotavirus genotypes were G9P[8] and G9P[4], Conclusion: Regarding the high frequency of rotavirus infection, continuous monitoring is needed in gastroenteritis prevention programs for treatment and to provide information about the occurrence of new rotavirus strains. [ABSTRACT FROM AUTHOR]

We report a rare case of necrotizing fasciitis of the abdominal wall caused by Hungatella effluvii and Streptococcus constellatus. Necrotizing fasciitis has high mortality, so early diagnosis and aggressive treatment are essential for good clinical outcome. Identification of the microbial contribution to these infections is crucial for targeted antibiotic treatment. • Necrotizing fasciitis needs early diagnosis and treatment due to its high mortality. • We report a rare case of necrotizing fasciitis due to Hungatella effluvii and Streptococcus constellatus. • Hungatella effluvii is an unusual pathogen isolated in human infections. • The isolates identified as Hungatella hathewayi by MALDI TOF MS, even with high score, may be Hungatella effluvii. [ABSTRACT FROM AUTHOR]

The aim of this study is to investigate the antitumor activity of Plantago major L. extract in Ehrlich ascites tumor (EAT) bearing Balb/C mice in vivo. Thirty male Balb/C mice were divided into 5 groups: 3 treatment groups and 2 control groups (6 per group). Treatment groups and the negative control group were injected with EAT (1 × 106 cells) intraperitoneally to develop ascites tumor. P. major L. extract (1%, 2% and 3% concentration extracts, 0.1 ml/day/mouse) were given p.o. for 10 alternate days. The control group was treated with 0.9% NaCl solution (0.1 ml/day/mouse). The changes of body weight in animals were recorded. On the 11th day, all of the mice were sacrified and their tissues were stained with haematoxylen and eosin for pathological studies. Body weights of in 3 treatment groups and the negative control group were elevated because of tumor burden. The maximal weight gain was recorded in the negative control group and the minimal weight gain was recorded in Group I. Pathological studies showed that P. major L. extract (especially 1% concentration) has inhibitive effect on EAT. P. major has an inhibitory effect on EAT in a dose dependent manner. [ABSTRACT FROM AUTHOR]