Carbapenem-resistant Klebsiella pneumoniae outbreak with monoclonal spread: Evaluation of resistance genes and ceftazidime-avibactam susceptibility.

PURPOSE: The aim of this study was to investigate ceftazidime-avibactam (CAZ-AVI) susceptibility, carbapenemase genes, and clonal relationship in carbapenem-resistant Klebsiella pneumoniae (CrKp) isolates. METHODS: A total of 28 non-repetitive CrKp isolates with positive carbapenemase production determined by the modified carbapenem inactivation method (mCIM), were included in the study. Identification of the isolates was performed with MALDI-TOF MS (VITEK-MS, bioMerieux, France). The automated system (VITEK-2, bioMerieux) and gradient diffusion test (Etest, bioMerieux) were used to determine antibiotic susceptibility. The mCIM was performed according to CLSI (2021) recommendations. CAZ-AVI susceptibility was carried out using the standard disc diffusion method. Results were evaluated according to EUCAST 2022 criteria. The blaOXA-48, blaNDM, blaKPC, blaIMP and blaVIM genes were investigated by multiplex PCR. The clonal relationship between isolates was determined by both AP-PCR and PFGE methods. RESULTS: Of the total 28 isolates, 89.3% were susceptible to CAZ-AVI. blaOXA-48 gene was found in 85.7% of the isolates, blaOXA-48+blaNDM gene in 10.7%, and blaNDM gene in 3.6%. blaKPC, blaIMP and blaVIM genes were not detected. Three clusters with three different genotypes were determined by the PFGE method. The largest cluster was Genotype A (n:24), followed by Genotype B (n:3), and Genotype C (n:1). AP-PCR was highly compatible with PFGE. The isolates of Genotype A, mostly from the intensive care unit (ICU), were evaluated as outbreak strains with monoclonal dissemination. CONCLUSIONS: OXA-48 remains the most frequently detected enzyme in CrKp strains in our country. The ceftazidime-avibactam susceptibility rate of 89.3% indicates that this antibiotic is still effective against CrKp isolates. The unnoticed outbreak detected in our study revealed the severity of intra-hospital cross-contamination affecting different wards, including the ICU. Therefore, in order to limit the spread of CrKp isolates, it is of great importance to implement strict infection control measures, and molecular surveillance programs, especially in the ICU.

[A Case of Mycobacterium abscessus Identified by Suspicious Gram Staining in the Blood Culture of a Patient with Chronic Renal Failure]. / Kronik Böbrek Yetmezlikli Hastanin Kan Kültüründe Süpheli Gram Boyamasi ile Tanimlanan Mycobacterium abscessus Olgusu.

Mycobacterium abscessus (M.abscessus), which is from the group of non-tuberculosis mycobacteria and is widely found in the natural environment, has been reported with increasing frequency as the causative agent of various infections; especially in the lower respiratory tract and in immuncompromised people. In this report, a case of M.abscessus, which developed tubular adenoma, pancytopenia and sepsis on the basis of chronic renal failure (CRF) was diagnosed by suspecting the causative agent in the Gram stain examination prepared from blood culture, was presented. A 49-year-old patient with CRF, who had complaints of weight loss, weakness, and loss of appetite for the last six months, admitted to the emergency department with a 7-8-day history of severe diarrhea and fever. Besides other tests, as the white blood cell count was 1.6 x 103/µl, neutrophil count was 80.6%, hemoglobin was 9.3 g/ dl and the platelet value was 36 x 103/µl in the blood samples, the patient was first taken into internal medicine service and then to the intensive care unit with a preliminary diagnosis of hypotension and sepsis. Meropenem and teicoplanin were started with the preliminary diagnosis of peritonitis in the internal medicine service. In addition to other tests, on the fifth day of antibiotic treatment, two consecutive sets of blood cultures were taken and sent to the microbiology laboratory. A positive signal was obtained from two aerobic blood culture samples at 42 and 45 hours of incubation in the BacT/Alert device. No bacteria were observed in the Gram staining of these samples and Erhlich Ziehl Neelsen (EZN) staining was performed because the structures considered as dye residues were noted as a result of the examination. Acid-fast bacteria were observed in the EZN-stained slide examination, and a panic report was given to the clinician. The patient died shortly after the notification was made in the evening hours. On culture plates inoculated after a positive signal, at the end of two days of aerobic incubation at 37 °C, small smooth S colonies grew on chocolate and sheep blood agar. Growing bacteria were detected as positive by EZN staining and identified as M.abscessus with 99.9% confidence by MALDI-TOF MS. After the bacterium was named as M.abscessus, the isolates were sent to the tuberculosis central laboratory of Süreyyapasa Chest Diseases and Thoracic Surgery Hospital for molecular typing. After DNA extraction from the growing colonies and polymerase chain reaction (PCR), they were typed using the GenoType NTM-DR (Hain Lifescience GmbH, Germany) kit and identified as M.abscessus, consistent with the MALDITOF MS result. After the species level identification, the erm, rrl (clarithromycin, azithromycin), and rrs (kanamycin, amikacin, and gentamicin) genes were investigated in the isolate, and it was determined that the bacteria were resistant to macrolides and sensitive to aminoglycosides. In the clinic, it should be noted that, non-tuberculous mycobacteria may play a role as an agent in immunocompromised people. On the other hand, it should be considered that non-tuberculosis bacteria may be the causative agent, with gram-positive bacilli appearing as stain residues or pale staining in Gram stains made from samples of such patients. As in this case, if the agent is seen as dye residue in blood culture Gram staining samples, it may be life-saving to suspect the agent and to report the result to the clinician accurately and quickly after EZN staining.

A new risk scoring system for early prediction of surgical need in patients with adhesive small bowel obstruction: a single-center retrospective clinical study.

Purpose: Cases of adhesive small bowel obstruction are a nuisance to surgeons. There have been years of ongoing discussions, and various guidelines have been published for the management of this disease. Both surgical and conservative approaches can have their own complications. It is often difficult to decide which treatment to apply to which patient. We aimed to create a multiparametric scoring system for the optimal management of adhesive small bowel obstruction patients. Methods: The retrospective laboratory, clinical and radiological records of 100 patients who were hospitalized and followed-up/treated for adhesive small bowel obstruction secondary to surgery in the General Surgery Clinic of Haydarpasa Numune Education and Research Hospital (Istanbul) between 2011 and 2021 were reviewed and statistically analyzed. Results: Admittance CRP and the largest diameter of the small intestine in the horizontal section of the admittance CT scans were significantly higher (P = 0.006 and P = 0.007), and the admittance albumin and sodium values were significantly lower (P < 0.001 and P = 0.031) in patients operated on for adhesive small bowel obstruction than in patients not operated on. Free intraperitoneal fluid in CT scans was detected at a higher rate in the operated group. An adhesive small bowel obstruction surgery score above 3.5 points out of 7 was found to be significant (P < 0.001). Conclusion: With this easy and applicable scoring system, complications of existing disease may be avoided by considering earlier surgical intervention in patients with a score of 4 and above.

[Evaluation of In vitro Efficacy of Meropenem/Colistin and Meropenem/Fosfomycin Combinations on Multidrug Resistant Gram-Negative Bacilli]. / Çok Ilaca Dirençli Gram-Negatif Basillerde Meropenem/Kolistin ve Meropenem/Fosfomisin Kombinasyonlarinin In vitro Etkinliginin Degerlendirilmesi.

The rate of extensively drug-resistant and pan-resistant gram-negative rods isolated as infectious agents is increasing around the world and in Türkiye. One of the important options in the treatment of these infections is the combined use of antibiotics. Therefore, the aim of this study was to investigate the in vitro effect of meropenem/colistin and meropenem/fosfomycin combinations on carbapenem-resistant gram-negative bacilli isolated as infectious agents. Escherichia coli (n= 6), Klebsiella pneumoniae (n= 10), Pseudomonas aeruginosa (n= 5), and Acinetobacter baumannii (n= 6) isolates were recovered from blood and tracheal aspirate samples of patients hospitalized in our hospital's intensive care unit were included in the study. In the first stage of the combination study, minimal inhibitory concentrations (MIC) were investigated by broth microdilution for meropenem and colistin, and agar dilution methods for fosfomycin. In the second stage of the study, synergy, partial synergy, indifference, and antagonistic effects were investigated with the checkerboard method for the meropenem/colistin combination and the agar dilution method for the meropenem/fosfomycin combination. The checkerboard results were interpreted as follows: fractional inhibitory concentration index (FICI) values ≤ 0.5 synergy, < 0.5-≤ 1 partial synergy, > 1-≤ 4 indifference and FIC values of > 4 antagonism. MIC values obtained in the study were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Of the 27 isolates studied with the broth microdilution method, 63% were found to be colistin-resistant and 37% susceptible. The MIC values of fosfomycin against Enterobacterales group bacteria were found to be in the range of 2-2048 mg/L. Two of the six E.coli isolates and nine of the 10 K.pneumoniae isolates were found to be resistant to fosfomycin (IV). The MIC values of ≥ 128 mg/L were found in all 11 non-fermentative gram-negative rods with intrinsic resistance to fosfomycin. In the combination of meropenem/ colistin, synergy and partial synergy were observed in 11 (40.7%) of 27 isolates, an indifference effect was observed in 13 (48.2%), and antagonistic effects were observed in three (11.1%) of the isolates. The synergy and partial synergy effects of this combination were 37.5% for Enterobacterales group bacteria, 50% for E.coli, and 30% for K.pneumoniae. Regarding the 11 non-fermentative gram-negative rods included in the study, 83.3% synergy and partial synergy was found in A.baumannii for the meropenem/colistin combination, while no synergy and partial synergistic effect was found in P.aeruginosa. Meropenem/fosfomycin synergy and partial synergy effects were 83.3% (5/6) for E.coli, 100% (8/8) for K.pneumoniae, 100% (6/6) for A.baumannii, and 25% (1/4) for P.aeruginosa. In all of the isolates studied, meropenem/fosfomycin combination was found to be more effective than the meropenem/colistin combination. It would be meaningful to support these data obtained in vitro with clinical efficacy results to be obtained as a result of the application of antibiotics in vivo, taking into account the pharmacokinetic and pharmacodynamic properties of the antibiotics used in this study.

The Effect of Total Laboratory Automation on Urine Culture Result Times in a Consolidated Laboratory.

BACKGROUND: The Provincial Health Directorate of Istanbul province (Turkey) established a consolidated laboratory network consisting of four regional central laboratories to reduce general laboratory costs and increase laboratory efficiency and quality in all affiliated hospitals. As part of the consolidation project, the Total Laboratory Automation (TLA) system was installed in the microbiology department of the ISLAB-2 central laboratory. In this study, the turnaround time (TAT) of urine samples of a satellite laboratory where the system was not installed and the ISLAB-2 central laboratory were compared to evaluate the effect of consolidation and the TLA. METHODS: The TAT values of all urine samples processed between March 2021, when the TLA was installed, and October 2021, were retrospectively examined in the laboratory information system. While the TLA was used for the processing and evaluation of samples in the ISLAB-2 central laboratory, manual methods were employed in the satellite laboratory. Both laboratories used MALDI-TOF MS (bioMérieux, France) for bacterial identification and VITEK 2 Compact (bioMérieux, France) for antibiotic sensitivity testing. Kruskal-Wallis test was used to compare TAT between the two laboratories. p < 0.05 was taken as the level of statistical significance. RESULTS: A total of 78,592 urine cultures (71,906 in the central laboratory and 6,686 in the satellite laboratory) were included in the study. Negative samples were reported in 23.5 hours in the central laboratory and 37.1 hours in the satellite laboratory and positive samples in 55 hours and 61.7 hours in the same laboratories, respectively. The mean TAT of both positive and negative urine cultures were found statistically significantly lower in the central laboratory than in the satellite laboratory (p < 0.0001). While 82% of negative urine cultures were completed within the first 24 hours in the central laboratory, only 17% were processed in the satellite laboratory. While 61% of positive samples were processed within the first 48 hours in the central laboratory, 38% were completed in the satellite laboratory. CONCLUSIONS: We assume that TLA has a positive effect on the diagnosis and treatment of patients, thanks to its contribution to standardization, efficiency, increased quality, and earlier reporting.

Investigation of in vitro efficacy of meropenem/polymyxin B and meropenem/fosfomycin combinations against carbapenem resistant Klebsiella pneumoniae strains.

The incidence of infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) is increasing worldwide, and very limited number of effective antibiotics are available for therapy. In our study, the in vitro efficacy of meropenem/polymyxin B and meropenem/fosfomycin combinations against CRKP strains was investigated. The efficiency of meropenem/polymyxin B and meropenem/fosfomycin combinations was tested by checkerboard microdilution and checkerboard agar dilution methods, respectively, against 21 CRKP strains containing major carbapenem resistant genes (7 blaKPC, 7 blaOXA-48 gene, and 7 blaOXA-48+ blaNDM), and seven additional CRKP strains without carbapenemase genes.Among the 28 CRKP strains, the meropenem/polymyxin B combination was synergistic in ten (35.7%), partially synergistic in 12 (42.8%), and indifferent in six (21.4%) isolates. The meropenem/fosfomycin combination was found to be synergistic in three isolates (10.7%), partially synergistic in 20 (71.4%), and indifferent in five (17.8%). In 21 strains containing carbapenem resistance genes, meropenem/polymyxin B and meropenem/fosfomycin combinations exhibited synergistic/partial synergistic effects in 15 (71.4%) and 16 (76.2%) strains, respectively, compared to 100% synergistic/partial synergistic efficiency in both combinations in seven strains free of carbapenemase genes. No antagonistic effect was detected in either combination.Regardless of presence or absence of carbapenem resistance genes, meropenem/polymyxin B and meropenem/fosfomycin combinations both demonstrated high synergistic and partial synergistic activity against 78.4% and 82.1% of CRKP strains, respectively. Also, they have no antagonistic effects and can be used successfully to prevent therapeutic failure with monotherapy, according to our in vitro studies.

The Prevalence of Positive Donor Corneoscleral Rim Culture and its Association with Ocular Infection After Transplantation.

Objectives: The aim of the study was to determine the prevalence of positive corneoscleral donor rim cultures and to report keratitis and endophthalmitis after keratoplasty. Methods: Eye bank records and medical records of patients who underwent keratoplasty between September 1, 2015, and December 31, 2019, were retrospectively reviewed. Patients who had routine donor-rim culture taken during surgery and followed up for at least 1 year in the post-operative period were included in the study. Results: A total of 826 keratoplasty procedures were performed. A total of 120 (14.5%) cases had a positive donor corneoscleral rim culture. Positive bacterial cultures were obtained from 108 (13.7%) of the donors. Bacterial keratitis was observed in one patient (0.83% of recipients) who had a positive bacterial culture. Positive fungal cultures were obtained from 12 (1.45%) donors, of whom one (8.33% of recipients) developed fungal keratitis. Endophthalmitis was observed in one patient whose culture result was negative. Both bacterial and fungal culture results were similar in penetrating and lamellar surgical procedures. Conclusion: Although the donor corneoscleral rims have a high positive culture result, the rate of bacterial keratitis and endophthalmitis is low, the risk of infection is high in patients with a fungal positive donor rim. Closer follow-up of patients with fungal positive donor corneo-scleral rim result and initiation of aggressive antifungal treatment when infection occurs will be beneficial.

Evaluation of Broth Disk Elution Method to Determine Colistin Resistance in Klebsiella pneumoniae and Escherichia coli Strains.

BACKGROUND: The reference broth microdilution (rBMD) method for the determination of colistin resistance is very laborious and time consuming, and many manual errors can occur. There are also limitations in detection of colistin heteroresistance. Therefore, alternative methods with satisfactory performance are required for routine laboratory work. In our study, the colistin broth disk elution (CBDE) method recommended by the Clinical and Laboratory Standards Institute (CLSI) for the detection of colistin resistance in routine applications was compared with rBMD. The compatibility and error rates of the method were evaluated and its usability in routine laboratory studies was examined. METHODS: Eighty-nine multidrug resistant Klebsiella pneumoniae and five Echerichia coli strains isolated from various clinical specimens were included in the study. Identification of strains and antibiotic susceptibility tests were performed with MALDI-TOF MS (bioMerieux, France) and Vitek-2 (bioMerieux) system. Minimum inhibitory concentration (MIC) was studied in 0.125 - 128 mg/L dilution range by using polystyrene microplate and colistin sulfate salt according to ISO-standard (20776-1) recommendations for the reference BMD test. The CBDE method was performed according to the CLSI recommendations. Isolates with MIC ≤ 2 mg/L were considered susceptible, while isolates with MIC > 2 mg/L were considered resistant according to EUCAST recommendations. The performance of the CBDE method was evaluated according to ISO criteria (Categorical agreement > 90%; major error and very major error rates < 3%). RESULTS: Categorical agreement for all 58 and 36 isolates found to be resistant and susceptible, respectively, by rBMD was found to be 100% with CBDE test. Since < 1 and > 4 µg/mL values could not be determined with the CBDE method, essential agreement (EA) could not be calculated. No major or very major errors were detected. CONCLUSIONS: Our results showed that the performance of the CBDE test is good when compared to the rBMD method. According to our data, we believe that the CBDE method can be used in routine laboratories for the detection of colistin resistance.

Comparison of performance of LIAISON SARS-CoV-2 antigen assay with RT-PCR during the Omicron wave.

Due to the newly emerging Omicron variant, there is a need to re-evaluate the performance of automated antigen tests. Our study aim was to evaluate the performance of the automated Liaison SARS-CoV-2 antigen assay against reverse transcriptase polymerase chain reaction (RT-PCR) in samples with Omicron variant.A prospective study was performed on 373 combined oro-nasopharyngeal samples (NPS) randomly collected from symptomatic patients. NPS were tested with Liaison SARS-CoV-2 Ag test (DiaSorin, Italy) and DS Coronex COVID-19 Multiplex RT-PCR Diagnosis Kit (DS BioTechnology, Ankara, Turkey).Of 373 samples, 124 (33.2%) were found to be RT-PCR positive and 249 (66.8%) RT-PCR negative. Taking RT-PCR as a reference, the sensitivity and specificity of the Liaison SARS-CoV-2 Ag assay were found as 84.6% (95%CI 77.3%-90%) and 100% (95%CI 98.5%-100%), respectively. For samples with a cycle threshold (Ct) value <25 (high viral load), the sensitivity increased to 100%. When antigen concentration and Ct values were compared, a strong negative correlation between antigen and Ct values was determined (P < 0.001).The Liaison antigen test met the performance criteria recommended by the WHO for samples with the Omicron variant. In addition, it showed excellent sensitivity and specificity in patients with high viral load. Therefore, Liaison antigen test can be a reliable and useful alternative in the diagnosis of SARS-CoV-2 infection, particularly in resource-constrained laboratories.

Risk factors for carbapenem-resistant Klebsiella pneumoniae infections in Intensive Care Units: a multicentre case-control study with a competing-risks analysis.

Aim: This study investigated the risk factors for the development of carbapenem-resistant Klebsiella pneumoniae (CRKP) infections in adult patients in Intensive Care Units (ICUs). Methods: A multicentre case-control study was conducted in ICUs in three tertiary hospitals in Turkey. The cases were patients culture-confirmed CRKP and a condition associated with healthcare-associated infections. Two controls were randomly selected for each case from among all other patients with an ICU stay at least as long as that of the corresponding case-patient. A proportional semiparametric subdistribution hazards regression model was used to assess risk factors for CRKP infection. ICU discharge and non-CRKP-related deaths were treated as competing risks. Results: A total of 120 patients, 44 cases and 76 controls were included in the analysis. Of the controls, 32 were discharged from the ICU and 44 died without acquiring CRKP infection. Endotracheal intubation (hazard ratio [HR]: 1.96, 95% confidence interval [CI]: 1.00-3.868) and type 2 diabetes mellitus (HR: 1.57, 95% CI: 0.888-2.806) were associated with an increased risk of CRKP infection, whereas carbapenem exposure (HR: 0.47, 95% CI: 0.190-1.1175) and the presence of a nasogastric tube (HR: 0.49, 95% CI: 0.277-0.884) were associated with a decreased risk of CRKP infection. Conclusions: Enteral nutrition support via a nasogastric tube may be associated with a reduced risk of CRKP-resistant infections in ICU patients. This hypothesis should be tested with a well-designed study.

Distribution of respiratory viruses: Evaluation of multiplex PCR results of 3074 patients.

OBJECTIVE: Early and accurate diagnosis of acute respiratory infections is important because these diseases negatively affect public health and can lead to loss of workforce and an increase in health expenditures. In this study, we aimed to determine the respiration panel multiplex polymerase chain reaction (PCR) test results and seasonal distribution in our region. METHODS: Three thousand and seventy-four patients samples multiplex PCR (Anatolia, Bosphore® Respiratory Pathogens Panel Kit v1) test results, which were sent to our laboratory, from 13 hospitals in our region between January 2018 and December 2018, were evaluated retrospectively. RESULTS: A total of 3074 patients samples, 1465 (48%) were positive and 1609 (52%) were negative test results. The most common factors were rhinovirus 30.2%, influenza A 23.1%, and respiratory syncytial virus (RSV) A/B 19.1%, respectively. When the distribution of these three most common viruses by months is examined, the most frequent months were determined as June for rhinovirus, November for influenza A, and February for RSV A/B. In the period between October and February, there was a significant increase in the positivity level of viral factors. CONCLUSION: The use of molecular methods in the diagnosis of respiratory infections will prevent unnecessary use of antibiotics and ensure correct and rapid treatment.

Cardiac surgery with cardiopulmonary bypass markedly lowers SARS-COV-2 antibody titer.

Background: This study aims to investigate the effect of cardiopulmonary bypass on antibody titers in patients vaccinated against the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) undergoing cardiac surgery with cardiopulmonary bypass. Methods: Between October 2021 and October 2022, a total of 70 patients (44 males, 26 females; mean age 59.9±10.3; range, 26 to 79 years) who completed their recommended COVID-19 vaccinations and underwent elective cardiac surgery with cardiopulmonary bypass were prospectively included. Serum samples for antibody titer measurements were taken at anesthesia induction and the end of cardiopulmonary bypass after decannulation. The SARS-CoV-2 total immunoglobulin antibodies against N-protein were measured. The antibody titer measurements at anesthesia induction and at the end of cardiopulmonary bypass were compared in all patients. Results: The median levels after cardiopulmonary bypass were lower than the preoperative levels (1,739.0 vs. 857.0, respectively; p<0.001). There was a drop of 40.0% (21.2%-62.6%) in the antibody titers among all patients. The decrease in antibody titers was consistent regardless of the number of vaccine doses or whether the last dose was received within the last three months. Among the studied factors, no parameter was significantly associated with a lesser or higher decrease in antibody titers. Conclusion: Cardiac surgery with cardiopulmonary bypass causes a decrease in SARS-CoV-2 antibody titers at the end of cardiopulmonary bypass. Revaccination after cardiac operations may be considered in this patient group that is highly vulnerable due to their comorbidities and lowered antibody levels.

[One-Year Candida Data of the Central Mycology Laboratory: Which Sample, Which Species, How Resistant?] / Merkez Mikoloji Laboratuvarinin Bir Yillik Candida Verileri: Hangi Örnek, Hangi Tür, Ne Kadar Dirençli?

The incidence of fungal infections particularly Candida species, is increasing gradually as a result of the increased life expectancy associated with the advances in the diagnosis and treatment of diseases, and increased number of patients in the risk group over the years. In addition, the incidence of fungal infection types that are resistant to antifungal drugs has been increasing, and rare fungal species have been reported to be isolated more frequently. For this reason, it is indicated that identification to the species level will contribute to the early initiation of an accurate and effective treatment. In this study, it was aimed to define the Candida species isolated from various clinical specimens and to document the performance of antifungal sensitivity tests. The Candida isolates sent to the central mycology laboratory in 2019 for identification and antifungal susceptibility tests were included in the study. The definition of the fungi to the species level was carried out using matrix-assisted laser desorption ionization-time of fl ight mass spectrometry (MALDI-TOF MS) and conventional methods. In vitro antifungal drug susceptibilities were analyzed using the The Clinical and Laboratory Standarts Institute (CLSI, M27-A3) reference broth microdilution method. The minimum inhibitory concentration (MIC) results were interpreted in accordance with the species-specific clinical breakpoints (CBPs) cited in the CLSI-M60 guidelines, and according to the epidemiological cut-off value (ECV) when no CBP was mentioned. The distribution of the species of the total 813 Candida isolates included in the study were as follows: Candida albicans (n= 312 ), Candida parapsilosis (n= 202), Candida tropicalis (n= 92), Candida glabrata (n= 71), Candida dubliniensis (n=28), Candida lusitaniae (n= 26), Candida kefyr (n= 22), Candida utilis (n= 17), Candida krusei (n= 14), Candida orthopsilosis (n= 7), Candida inconspicua (n= 7), Candida guilliermondii (n= 5), Candida metapsilosis (n= 4), Candida norvegensis (n= 4), Candida lambica (n= 1) and Candida lipolytica (n= 1). The evaluation of the results of the antifungal susceptibility tests according to the CBPs revealed that one C.albicans isolate and 60 C.parapsilosis (29.7%) isolates were resistant, and seven C.parapsilosis (3.5%) isolates were dose-dependent susceptible to fluconazole; 32 C.parapsilosis (15.8%) isolates were intermediately susceptible to voriconazole; one C.parapsilosis (0.5%) was resistant and one C.krusei (7.1%) was intermediately susceptible to anidulafungin; and one C.parapsilosis (0.5%) was resistant and one C.krusei (7.1%) isolate was intermediately susceptible to micafungin. In terms of ECVs, one C.lusitaniae isolate for fluconazole and one of each C.lusitaniae and C.kefyr isolates were evaluated as a non-wild type. In the present study, 61 of 813 isolates were found to be resistant to fluconazole and seven were dose dependently susceptible, 32 were intermediately susceptible to voriconazole, one was resistant to anidulafungin, one was intermediately susceptible, and one was resistant to micafungin and one was intermediately susceptible to micafungin. In conclusion, the increased number of non- albicans Candida species and increased levels of resistance to antifungal drugs further establish the importance of early diagnosis at a species level alongside antifungal susceptibility tests.

Performance of the Rapidec®Carba NP assay for the detection of different carbapenemases in Klebsiella pneumoniae and Escherichia coli strains.

PURPOSE: The spread of infections caused by Enterobacterales strains resistant to carbapenems is a global public health problem, and early detection of carbapenemases is very important to prevent their spread. The rapid detection of carbapenemase production with the new commercial assay Rapidec® Carba NP test is based on the biochemical detection of imipenem hydrolysis. Our study aims to evaluate the performance of the Rapidec® Carba NP test in OXA-48 positive isolates highly prevalent in our country and also in isolates with more than one carbapenemase gene that have an increased prevalence and to examine whether it can be used for confirmation of carbapenemase positivity in the routine laboratory. METHODS: A total of 97 strains of 94 carbapenem-resistant Klebsiella pneumoniae and three carbapenem-resistant Escherichia coli isolated from various clinical specimens were included in the study. The results of the Rapidec® Carba NP assay were compared with those obtained by the multiplex PCR test. RESULTS: The sensitivity of the Rapidec® Carba NP test was 97.8% for all carbapenemase-positive isolates. Of 90 PCR positive isolates, one OXA-48 and one OXA-48 â€‹+ â€‹NDM positive isolates were negative with Rapidec® Carba NP test. CONCLUSIONS: The positive results detected by the Rapidec® Carba NP test make an important contribution to the early detection of carbapenemase production and infection control practices. Since two carbapenemase positive isolates were found to be negative with the Rapidec® Carba NP test in our study, it was concluded that negative results of carbapenem-resistant isolates obtained with this assay should be confirmed with an additional carbapenemase detection method to exclude false-negative results.

Microorganisms isolated from the bile of the patients who have undergone cholecystectomy and their antibiotic resistance pattern: multicenter prospective study.

BACKGROUND: Gallbladder and biliary tract infections are diseases with high mortality rates if they are not treated properly. Microbiological evaluation of perioperatively collected samples both ensures proper treatment of patients and guides empirical treatment due to the determination of microorganism susceptibility. AIMS: This study aimed to isolate the microorganisms in bile cultures from patients who underwent cholecystectomy and to determine sensitivity results of these microorganisms. METHODS: This study was a multi-center and prospective design, included 360 patients, and was performed between 2019 and 2020. Culture results of bile taken during cholecystectomy were evaluated. RESULTS: Bacterial growth was found in the bile cultures of 84 out of 360 (23.3%) patients. Patients were divided into two groups according to whether they had risk factors for resistant microorganisms or not. While Escherichia coli (n = 11, 13%), Enterococcus spp. (n = 8, 9.5%), and Enterobacter spp. (n = 4, 4.7%) were detected most frequently in patients without risk. Staphylococcus spp. (n = 17, 20.2%), Enterococcus spp. (n = 16, 19%), and E. coli (n = 8, 9.5%) were the most frequently found microorganism at-risk patients. In multivariate analysis, bile culture positivity was found higher in patients who had history of biliary disease (p = 0.004), operation performed concurrently with a cholecystectomy (p = 0.035), and high rate of polymorphonuclear leukocytes (PNL) in total leukocyte count (p = 0.001). CONCLUSIONS: Our study shows that when starting empirical antibiotic treatment for bile ducts, whether patients are at risk for the development of resistant bacterial infection should be evaluated after which antibiotic selection should be made accordingly.

Phenotypic detection of carbapenemase production in carbapenem-resistant isolates with the rapid carbapenemase detection method (rCDM).

INTRODUCTION: Carbapenem antibiotics are widely used for the treatment of infections caused by multidrug-resistant bacteria. As a result of this, resistance to carbapenems is gradually increasing. Identification of carbapenemase production, one of the reasons for resistance, through molecular methods is expensive and time-consuming. In the present study, it was aimed to investigate the sensitivity of the newly developed rapid carbapenemase detection method (rCDM) as compared to the gold standard molecular method and to evaluate its consistency with another phenotypical method, the modified carbapenem inactivation method (mCIM). MATERIAL AND METHODS: In our study, a total of 152 Gram-negative bacteria (Klebsiella pneumoniae, Escherichia coli) isolated from various clinical samples of which 50 were controls were included. Strain identification was done by using VITEK®MS (bioMérieux, Marcy-I'Étoile, France), carbapenem sensitivity was tested by using VITEK®2 (bioMérieux). For carbapenem-resistant isolates, carbapenem resistance genes were detected with multiplex PCR [Carbapenem and Colistin Resistance qPCR kit (Bioeksen, Istanbul, Turkey)] kit by a molecular method. All included isolates were evaluated by the rCDM and mCIM tests in order to detect carbapenemase phenotypically. The molecular method was accepted to be the gold standard and the sensitivity of rCDM was calculated. The McNemar test was applied to analyze the difference between two phenotypic tests (rCDM and mCIM) and Cohen's Kappa analysis was applied to determine consistency. RESULTS: Out of 102 carbapenem-resistant isolates, at least one of the resistance genes in the multiplex PCR panel (blaKPC, blaNDM, blaVIM, blaIMP, blaOXA-51, blaOXA23/58, blaOXA-48) was detected in 92 and blaOXA-48 was the most common (90.2%). The sensitivity of the rapid carbapenemase detection method was found to be 100%. When the results of the two phenotypic methods were compared, no statistically significant difference could be found (PMcNemar:1, Kappa coefficient:1.00). CONCLUSION: The rapid carbapenemase detection method was found to be suitable to use in routine laboratory analysis as its sensitivity was found to be high, exhibited a good performance for detection of frequent carbapenemase types in our country (Turkey), a high consistency with mCIM, and also it is an easily applied and rapid method.

Hepatitis E Infection in Solid Organ Transplant Recipients in Turkey.

BACKGROUND: Hepatitis E virus is a re-emerging pathogen with an increase in human cases that can lead to chronic infection in immunosuppressed patients. Turkey is located between Asia and Europe, 2 regions with distinct epidemiological and clinical features of hepatitis E virus infection. This multicenter cross-sectional study aimed to investigate the prevalence of hepatitis E virus infection in liver and kidney transplant recipients in Turkey and to determine the role of possible transmission factors. METHODS: A total of 485 plasma samples of solid organ recipients were collected from 7 transplantation centers in Turkey. Samples were tested for anti-hepatitis E virus immunoglobin M, immunoglobin G, and hepatitis E virus ribonucleic acid. Water- and food-related risk factors were evaluated by a questionnaire. RESULTS: Samples of 300 kidney and 185 liver recipients were collected. Hepatitis E virus ribonucleic acid was tested in 472 samples and none were positive. Anti-hepatitis E virus immunoglobin G and immunoglobin M were detected in 84 (17.3%) and 3 (0.6%) patients, respectively. Seropositivity was associated with older age, male gender, being a liver recipient, and being infected with hepatitis B virus and/or hepatitis C virus. None of the patients under the age of 30 were seropositive. Hepatitis E virus immunoglobin G prevalence was higher in the Central East and Southeast Anatolia. Eating raw meat was the only independent variable associated with hepatitis E virus seropositivity. CONCLUSION: This is the first prevalence study of hepatitis E virus infection in solid organ recipients in Turkey. Anti-hepatitis E virus immunoglobin G prevalence was 17.3% which was higher than the previously reported rate in blood donors. Seropositivity was significantly higher in liver recipients. Despite the high antibody prevalence, none of the patients were viremic.

Comparison of syphilis seropositivity between non-immigrant and immigrant populations in the Anatolian side of Istanbul, Turkiye: Results of five-years retrospective study.

OBJECTIVE: Our study aimed to evaluate the seropositivity for syphilis in non- immigrant and immigrant populations and compare the results regarding demographic data. METHODS: In accordance with the reverse algorithm, syphilis tests were performed between May 2014 and December 2018 in hospitals in our service zone for syphilis screening or symptomatic disease. RESULTS: A total of 135.328 non- immigrant and 6.641 immigrant were screened for syphilis. Seropositivity rates were 1.3% in the non- immigrant and 3.8% in immigrant groups (p=0.0001). There was a statistically significant difference in terms of seropositivity rates between the various age groups in the local group and immigrant groups (except 18 25 age group) (p<0.05). Syphilis seropositivity rates were found to be lower in indigenous population than immigrant groups according to the years tested (p=0.0001). The seropositivity rates were 2.4% and 3.2% among the males (p=0.025) and 0.6% and 4.0% among females (p=0.0001) in non-immigrantand immigrant groups, respectively. Whereas, 0.6% of pregnant women in the local group and 3.7% of pregnant women in immigrant groups were seropositive for syphilis (p=0.0001). Among the HIV positive group, syphilis seropositivity was only observed in the non-immigrant group with a rate of 23.0% (p=0.0001). CONCLUSION: The antibodies against syphilis were found more frequently in immigrants than non-immigrant. Among the HIV positive individuals syphilis seropositivity was only observed in the non-immigrant group.

Bartonella henselae IgM seropositivity in both adult and pediatric patients with diverse clinical conditions in Turkey.

Bartonella henselae is the causative agent of cat scratch disease (CSD). In this study, we aimed to investigate the clinical data of patients with suspicion of CSD and delineate current epidemiological features.A total of 785 patients with suspected CSD were included in the study. B. henselae IgM antibody was determined by indirect fluorescent antibody (IFA) test using a commercial kit (Euroimmun, Germany). Sex, age, clinical pre-diagnosis and animal contact information of the patients were obtained from hospital electronic database records.Seventy-eight (9.9%) of 785 samples were seropositive. Out of 78 patients, 46 with animal contact data were further analyzed. Of these patients, 56% were male, and 41% were under 18 years of age. Seropositivity was more commonly observed in fall and winter. The most common finding was lymphadenitis (63%). Thirty-five patients (76%) had a previous history of animal contact (cat/dog). Of the 46 seropositive patients, 78.3, 15.2, 4.4, and 2.1% had titers of 1:80, 1:160, 1:320, and 1:640, respectively.Our study confirms that CSD is not rare in Turkey. Thus, it should always be considered in the differential diagnosis of patients presenting with lymphadenopathy in all age groups, particularly children. Questioning of cat exposure should never be neglected, especially in areas with intense population of stray cats, such as Istanbul.

Detection of colistin resistance among multidrug-resistant Klebsiella pneumoniae and Escherichia coli clinical isolates in Turkey.

In this study investigation of plasmid-mediated mcr 1-5 resistance genes was performed among multidrug-resistant (MDR) colistin sensitive and resistant Klebsiella pneumoniae and Escherichia coli strains isolated in our laboratory. We aimed to evaluate automated system (Vitek-2), broth microdilution (BMD) reference method and chromogenic media performance. Totally 94 MDR K. pneumoniae and six E. coli isolates were included in the study. CHROMID® Colistin R agar (COLR) (bioMerieux, France) was used to determine the colistin resistance by chromogenic method. Standard PCR amplification was performed using specific primers to screen the plasmid-mediated mcr 1-5 genes. Sixty-one isolates were resistant to colistin and 39 were susceptible with reference BMD. The essential and categorical agreement of Vitek-2 was determined as 100 and 99%. The sensitivity of COLR medium was 100%, the specificity was 97.5%. In our study mcr-1 was detected in eight isolates, while other mcr genes were not detected. Due to the high sensitivity and specificity of the COLR medium, it can be used in routine diagnostics for the detection of colistin resistance. In our study we detected 8% prevalence of mcr-1 among MDR strains however, two mcr-1 positive isolates were found sensitive to colistin by BMD.