Detection of VIM, NDM and OXA-48 producing carbapenem resistant Enterobacterales among clinical isolates in Southern Hungary.

Infections caused by carbapenem-resistant Enterobacterales (CRE) present an important therapeutic problem, as there are limited number of effective therapeutic alternatives available. In this study, phenotypic and genotypic methods were used to characterize carbapenemase-production and other resistance-determinants (AmpC and ESBL-production, efflux pump-overexpression) in 50 isolates (Klebsiella spp. n = 35, Escherichia coli n = 12 and Enterobacter cloacae complex n = 3) collected at the Albert Szent-Györgyi Clinical Center (University of Szeged) between 2014 and 2017. Minimum inhibitory concentrations of meropenem, sulfamethoxazole/trimethoprim, tigecycline, amikacin, moxifloxacin, colistin and fosfomycin were also determined. 24% of isolates were AmpC-producers, while 30% carried blaCTX-M ESBL-genes. Carbapenemase-genes were detected in 18 (36%) of the tested isolates: in 2 isolates blaNDM, in 6 isolates blaOXA-48-like and in 12 isolates, blaVIM was detected by PCR. The species-distribution for isolates positive for carbapenemase-genes was the following: Klebsiella pneumoniae n = 11, Klebsiella oxytoca n = 1, E. coli n = 5, E. cloacae complex n = 1. Efflux pump-overexpression based on the PAßN-screening agar was shown in n = 3 of the tested strains. In nine isolates (18%), carbapenemase and ESBL-genes were detected simultaneously. Highest levels of resistance were noted for fosfomycin (74%) and moxifloxacin (70%), while all isolates were susceptible to colistin. Among applied phenotypic tests in this study the modified carbapenem inactivation method (mCIM) proved to be the most accurate one compared to that of PCR results.

Anaerobic blood culture positivity at a University Hospital in Hungary: A 5-year comparative retrospective study.

Anaerobic bacteremia (AB) is usually detected in about 0.5-13% of positive blood cultures. The aim of this study was to determine prevalence of anaerobic bacteremia over a 5-year period (2013-2017), to identify current trends at our University Hospital and to compare the results to those in a similar study (2005-2009) in the same region. During the study period, an average of 23,274 ±â€¯2,756 blood cultures were received per year. Out of the positive blood cultures, 3.3-3.6% (n = 423) yielded anaerobic bacteria, representing 3.5-3.8 anaerobic isolates/1000 blood culture bottles (including both aerobic and anaerobic bottles) per year for hospitalized patients. Mean age of affected patients was 70-73 years (range: 18-102 years) with a male-to-female ratio: 0.60. Most isolated anaerobes were Cutibacterium spp. (54.0 ±â€¯8.5%; n = 247), while among anaerobes other than Cutibacterium spp., Bacteroides and Parabacteroides and Clostridium spp. were the most prevalent. Blood culture time-to-positivity (TTP) for clinically relevant bacteria was 31.4 ±â€¯23.4 h, while for Cutibacterium spp., TTP values were 112.9 ±â€¯37.2 h (p < 0.0001). In conclusion, the prevalence of anaerobic bacteremia should be determined on institutional basis.

Comparative Epidemiology and Resistance Trends of Common Urinary Pathogens in a Tertiary-Care Hospital: A 10-Year Surveillance Study.

Background and Objective: Urinary tract infections (UTIs) are common in human medicine, affecting large patient populations worldwide. The principal cause of UTIs is uropathogenic Escherichia coli (UPEC) and Klebsiella, both in community and nosocomial settings. The assessment of local data on prevalence and resistance is essential to evaluate trends over time and to reflect on the national situation, compared to international data, using the methods of analytical epidemiology. Materials and Methods: The aim of this study was to assess resistance trends and epidemiology of UTIs caused by E. coli and Klebsiella species in inpatients and outpatients at a tertiary-care hospital in Hungary, using microbiological data. To evaluate resistance trends, several antibiotics were chosen as indicator drugs, based on local utilization data. Results: E. coli was the most prevalent isolate, representing 56.75 ± 4.86% for outpatients and 42.29 ± 2.94% for inpatients. For E. coli, the ratio of resistant strains for several antibiotics was significantly higher in the inpatient group, while in Klebsiella, similar trends were only observed for gentamicin. Extended-spectrum ß-lactamase (ESBL)-producing isolates were detected in 4.33-9.15% and 23.22-34.22% from outpatient, 8.85-38.97% and 10.89-36.06% from inpatient samples for E. coli and Klebsiella, respectively. Conclusions: Resistance developments in common UTI pathogens (especially to fosfomycin, sulfamethoxazole-trimethoprim, fluoroquinolones, and 3rd generation cephalosporins), seriously curb therapeutic options, especially in outpatient settings.

Recurrent Scedosporium apiospermum mycetoma successfully treated by surgical excision and terbinafine treatment: a case report and review of the literature.

BACKGROUND: Scedosporium apiospermum is an emerging opportunistic filamentous fungus, which is notorious for its high levels of antifungal-resistance. It is able to cause localized cutaneous or subcutaneous infections in both immunocompromised and immunocompetent persons, pulmonary infections in patients with predisposing pulmonary diseases and invasive mycoses in immunocompromised patients. Subcutaneous infections caused by this fungus frequently show chronic mycetomatous manifestation. CASE REPORT: We report the case of a 70-year-old immunocompromised man, who developed a fungal mycetomatous infection on his right leg. There was no history of trauma; the aetiological agent was identified by microscopic examination and ITS sequencing. This is the second reported case of S. apiospermum subcutaneous infections in Hungary, which was successfully treated by surgical excision and terbinafine treatment. After 7 months, the patient remained asymptomatic. Considering the antifungal susceptibility and increasing incidence of the fungus, Scedosporium related subcutaneous infections reported in the past quarter of century in European countries were also reviewed. CONCLUSIONS: Corticosteroid treatment represents a serious risk factor of S. apiospermum infections, especially if the patient get in touch with manure-enriched or polluted soil or water. Such infections have emerged several times in European countries in the past decades. The presented data suggest that besides the commonly applied voriconazole, terbinafine may be an alternative for the therapy of mycetomatous Scedosporium infections.

[Special application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry in clinical microbiological diagnostics]. / Mátrix-asszisztált lézer deszorpciós, ionizációs, repülési ido mérésén alapuló tömegspektrometria speciális alkalmazása a klinikai mikrobiológiai diagnosztika területén.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry as a new possibility for rapid identification of bacteria and fungi revolutionized the clinical microbiological diagnostics. It has an extreme importance in the routine microbiological laboratories, as identification of the pathogenic species rapidly will influence antibiotic selection before the final determination of antibiotic resistance of the isolate. The classical methods for identification of bacteria or fungi, based on biochemical tests, are influenced by many environmental factors. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a rapid method which is able to identify a great variety of the isolated bacteria and fungi based on the composition of conserved ribosomal proteins. Recently several other applications of the method have also been investigated such as direct identification of pathogens from the positive blood cultures. There are possibilities to identify bacteria from the urine samples in urinary tract infection or from other sterile body fluids. Using selective enrichment broth Salmonella sp from the stool samples can be identified more rapidly, too. The extended spectrum beta-lactamase or carbapenemase production of the isolated bacteria can be also detected by this method helping the antibiotic selection in some cases. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry based methods are suitable to investigate changes in deoxyribonucleic acid or ribonucleic acid, to carry out rapid antibiotic resistance determination or other proteomic analysis. The aim of this paper is to give an overview about present possibilities of using this technique in the clinical microbiological routine procedures.

Urinary Tract Infections in Elderly Patients: A 10-Year Study on Their Epidemiology and Antibiotic Resistance Based on the WHO Access, Watch, Reserve (AWaRe) Classification.

The ageing of the population-especially in developed countries-has brought on many societal challenges and has significantly contributed to the burden on healthcare infrastructures worldwide. Elderly persons (aged ≥ 65 years) are at higher risk for developing UTIs, due to a range of intrinsic and extrinsic risk factors, and they often delay seeking treatment. A retrospective observational study was performed regarding the epidemiology and resistance of UTIs in elderly patients. Identification of the isolates was carried out using VITEK 2 ID/AST and MALDI-TOF mass spectrometry. Antibiotic resistance in these isolates was assessed based on EUCAST guidelines, and were grouped into the WHO AWaRe (Access, Watch, Reserve) classification of antimicrobials. During the 10-year study period, n = 4214 (421.4 ± 118.7/year) and n = 4952 (495.2 ± 274.6) laboratory-confirmed UTIs were recorded in inpatients and outpatients, respectively. The causative agents showed differentiation among outpatients and inpatients: Escherichia coli (48.14% vs. 25.65%; p = 0.001), Enterococcus spp. (20.15% vs. 21.52%; p > 0.05), Klebsiella spp. (16.28% vs. 16.26%; p > 0.05), Pseudomonas spp. (4.40%vs. 13.36%; p = 0.001); Proteus-Providencia-Morganella group (4.56% vs. 10.96%; p = 0.001); Candida spp. (0.53% vs. 5.98%; p = 0.001); Citrobacter-Enterobacter-Serratia group (1.90% vs. 2.71%; p < 0.05). Significantly higher resistance rates were observed in inpatient isolates for many Access and Watch antibiotics compared to isolates of outpatient origin; in addition, resistance rates were higher in these uropathogens compared to the previously recorded rates in the region. More care should be taken for the diagnosis and treatment of UTIs affecting elderly patients, as they represent a particularly vulnerable patient population.

Beta-Haemolytic Group A, C and G Streptococcal Infections in Southern Hungary: A 10-Year Population-Based Retrospective Survey (2008-2017) and a Review of the Literature.

INTRODUCTION: Pyogenic ß-hemolytic streptococci (including Group A, C and G Streptococcus) are some of the most important Gram-positive bacterial pathogens in human medicine. Although effective therapy is available, invasive streptococcal infections are associated with a significant disease burden. METHODS: In this retrospective study, the epidemiological characteristics of invasive Group A (iGAS) and Group C and G (iGCGS) streptococci, along with tonsillo-pharyngitis-causing pGAS and pGCGS infections, were assessed in Southern Hungary. A total of 1554 cases of streptococcal tonsillo-pharyngitis infections (26.5-44.1/100,000 persons, pGAS: 95.5%; n=1484) and 1104 cases of invasive streptococcal infections were detected (12.5-31.4/100,000 persons, iGAS: 77.9%; n=861). RESULTS: The average age of the affected patients in the various groups were the following: pGAS: 13.2±13.1 years, pGCGS: 21.0±15.0 years (p=0.039), iGAS: 49.1±12.8 years, iGCGS: 58.7±18.5 years (p>0.05). iGAS isolates originated from abscesses (47.1%), blood culture samples (24.1%), surgical samples (16.7%), biopsies (4.6%), pleural fluid (3.5%), pus (2.0%), synovial fluid (1.3%) and cerebrospinal fluid samples (0.7%). In contrast, iGCGS isolates mainly originated from blood culture samples (53.8%), abscesses (22.9%), surgical samples (12.3%), synovial fluid (5.1%), pleural fluid (3.7%), pus (1.8%) and cerebrospinal fluid samples (0.4%). All respective isolates were susceptible to benzyl-penicillin; overall resistance levels for erythromycin (10.5% for GAS, 21.4% for GCGS) and clindamycin (9.2% for GAS, 17.2% for GCGS) were significantly higher in GCGS isolates, while resistance levels for norfloxacin were higher in GAS isolates (13.5% for GAS, 6.9% for GCGS). CONCLUSION: The rates of resistance to macrolides and clindamycin are a cause for concern (especially among GCGS isolates); however, resistance levels are still relatively low, compared to Southern European countries.

Differential epidemiology and antibiotic resistance of lactose-fermenting and non-fermenting Escherichia coli: Is it just a matter of taste?

Urinary tract infections (UTIs) are some of the most common infections affecting humans worldwide. Occurrence of atypical, lactose non-fermenting, biochemically "inactive" strains of E. coli in clinical material has been described in the literature, which may cause a significant diagnostic challenge. The present retrospective microbiological study was carried out using isolates and data collected between January 1, 2013, and December 31, 2017, at the Institute of Clinical Microbiology. n = 24,285 positive urine samples were noted during the study period, out of which, samples positive for either lac + and lac- E. coli were included in the analysis. E. coli represented n = 7075 (55.8% ± 4.6%) of outpatient and n = 4916 (42.4% ± 3.6%) of inpatient isolates. n = 401 (3.3%; 80.2 ± 14.6/year) lac- E. coli isolates were identified from urinary tract infections. The ratio of lac- E. coli isolates was significantly higher in outpatient samples (262 vs. 139). Resistance levels of lac- isolates for antibiotics commonly used for treating UTIs were significantly higher for both inpatient and outpatient isolates: norfloxacin, ciprofloxacin, fosfomycin and nitrofurantoin. It is essential to pay attention to the presence of lac- strains, and their omission from clinical material during diagnostic procedures may have significant consequences for epidemiological studies and therapy.

Increasing relevance of Gram-positive cocci in urinary tract infections: a 10-year analysis of their prevalence and resistance trends.

Urinary tract infections (UTIs) are the third most common types of infection in human medicine worldwide. There is increasing appreciation for the pathogenic role of Gram-positive cocci (GPC) in UTIs, as they have a plethora of virulence factors, maintaining their pathogenicity and high affinity for the epithelial cells of the urinary tract. The study was carried out using microbiological data collected corresponding to the period between 2008 and 2017. Antimicrobial susceptibility testing was performed using the disk diffusion method and E-tests. The age range of patients affected from the outpatient and inpatient groups differed significantly (43 [range 0.7-99] vs. 68 [range 0.4-99] years; p = 0.008). 3962 GPCs were obtained from inpatient and 4358 from outpatient samples, corresponding to 20.5 ± 2.8% (range 17.5-26.8%) and 20.6 ± 2.6% (range 17.8-26.0%) of all positive urine samples (p > 0.05); in both groups, Enterococcus spp. were the most prevalent (outpatients: 79.6%; inpatients: 88.5%). High-level aminoglycoside resistance in enterococci was noted in 31.0-46.6% of cases. A pronounced increase in the number of MRSA was seen in the second half of the study period (0.6-1.9% vs. 9.8-11.6%; p = 0.038). The ratio of VRE isolates was 0.16%, no VISA/VRSA isolates were detected.

Leclercia adecarboxylata as an emerging pathogen in human infections: a 13-year retrospective analysis in Southern Hungary.

INTRODUCTION: The clinical role of Leclercia adecarboxylata as an opportunistic pathogen in the context of human infections have been highlighted by multiple published case reports, describing these bacteria as novel or emerging pathogens. METHODOLOGY: The study included L. adecarboxylata isolates and laboratory data collected, corresponding to a 13-year time period (between 1 January 2005 and 31 December 2017). Presumptively identified L. adecarboxylata isolates were re-identified using VITEK 2 Compact ID/AST and MALDI-TOF MS analysis. RESULTS: n = 34 isolates were verified by VITEK 2 system and MALDI-TOF. The fosfomycin-agar and CPS Elite agar were effective in the phenotypic differentiation of the isolates. N = 18 (52.9%) of L. adecarboxylata was considered as clinically significant pathogens (based on the clinical signs and symptoms), while n = 16 (47.1%) were considered as contaminants. These pathogens were isolated from wound/abscess samples (n = 9), urine samples (n = 6) and blood cultures (n = 3). 31 out of 34 isolates (91.2%) were pan-sensitive (i.e. wild type) to the tested antibiotics. The median age of affected patients was 57 years (range: 12-80 years), 11 out of 18 patients (61.1%) presented with underlying immunosuppression at the time of isolation. CONCLUSIONS: Based on the finding of this study, the actual (published) frequency of L. adecarboxylata infections needs to be re-evaluated as the risk of misidentification (and reporting the isolate as a pan-sensitive Escherichia coli) is high. Additional reporting of cases, both from a microbiological and clinical standpoint, could help clinicians develop a better understanding of the potential of this organism as a pathogen.

Characterization of Resistance in Gram-Negative Urinary Isolates Using Existing and Novel Indicators of Clinical Relevance: A 10-Year Data Analysis.

Classical resistance classifications (multidrug resistance [MDR], extensive drug resistance [XDR], pan-drug resistance [PDR]) are very useful for epidemiological purposes, however, they may not correlate well with clinical outcomes, therefore, several novel classification criteria (e.g., usual drug resistance [UDR], difficult-to-treat resistance [DTR]) were introduced for Gram-negative bacteria in recent years. Microbiological and resistance data was collected for urinary tract infections (UTIs) retrospectively, corresponding to the 2008.01.01-2017.12.31. period. Isolates were classified into various resistance categories (wild type/susceptible, UDR, MDR, XDR, DTR and PDR), in addition, two new indicators (modified DTR; mDTR and mcDTR) and a predictive composite score (pMAR) were introduced. Results: n = 16,240 (76.8%) outpatient and n = 13,386 (69.3%) inpatient UTI isolates were relevant to our analysis. Citrobacter-Enterobacter-Serratia had the highest level of UDR isolates (88.9%), the Proteus-Providencia-Morganella group had the highest mDTR levels. MDR levels were highest in Acinetobacter spp. (9.7%) and Proteus-Providencia-Morganella (9.1%). XDR- and DTR-levels were higher in non-fermenters (XDR: 1.7%-4.7%. DTR: 7.3%-7.9%) than in Enterobacterales isolates (XDR: 0%-0.1%. DTR: 0.02%-1.5%). Conclusions: The introduction of DTR (and its' modifications detailed in this study) to the bedside and in clinical practice will definitely lead to substantial benefits in the assessment of the significance of bacterial resistance in human therapeutics.

Epidemiology of candiduria and Candida urinary tract infections in inpatients and outpatients: results from a 10-year retrospective survey.

INTRODUCTION: The presence of Candida species in urine (candiduria) is a common clinical finding, which may frequently represent colonization or contamination of specimens, however, they may be etiological agents in urinary tract infections (UTIs) or be indicators of underlying pathology in the genitourinary system or disseminated candidaemia. C. albicans is the most frequently isolated species of the genus, however, an increase in the occurrence of non-albicans Candida species (NACS) has been reported, which may be attributable to frequent exposure to fluconazole. MATERIAL AND METHODS: The aim of this study was to retrospectively assess and compare the prevalence of candiduria and UTIs caused by Candida spp. among inpatients and outpatients at a major clinical center in Hungary, during a 10-year period (2008-2017). RESULTS: Candiduria was detected in 0.11-0.75% of positive samples from outpatients, while this number was significantly higher for inpatients, ranging between 3.49-10.63% (p <0.001). Overall, C. albicans was the most frequently isolated species (65.22% in outpatients and 59.64% in inpatients), however, the presence of C. glabrata as a relevant etiologic agent (~20-30%) is also noteworthy, because there are corresponding therapeutic consequences. CONCLUSIONS: A pronounced female dominance (1.7-2.15-fold), advanced age (~70 years) and hospitalization of affected patients during candiduria is in line with the findings in literature.

Evaluation of the Results of Group B Streptococcus Screening by MALDI-TOF MS among Pregnant Women in a Hungarian Hospital.

Pregnant women colonized by Streptococcus agalactiae, or group B streptococcus (GBS), are at an increased risk of premature delivery and stillbirth, and their neonates can be endangered by the development of an invasive GBS disease. In this study, the results of the GBS screening among pregnant women performed between 2012 and 2018 (n = 19267) are presented. For the GBS positive samples, the antibiotic susceptibility of the isolated strains was also tested (n = 3554). During the examined period, the colonization rate varied between 17.4% and 19.8%. The overall rate of erythromycin and clindamycin resistance in the GBS positive samples was 34.9% and 34.6%, respectively. The frequency of the erythromycin and clindamycin resistant strains showed an increasing tendency. An analysis of the MALDI-TOF MS spectra of 260 GBS isolates revealed that 46.5% of them belonged to either the ST-1 or the ST-17 sequence types, indicating a high prevalence of these potentially invasive GBS strains in our region. More than half of the strains identified as ST-1 (52.1%) proved to be resistant to erythromycin and clindamycin.

Combination of MALDI-TOF MS and PBP2' latex agglutination assay for rapid MRSA detection.

Sample preparation was optimized for MALDI-TOF MS directly from the selective enrichment broth to detect Staphylococcus aureus. A combination of MALDI-TOF MS and the PBP2' latex agglutination assay was applied for MRSA screening and evaluated on 255 clinical samples. MRSA colonisation can be reported already 18-24h after sample collection.

Combination of selective enrichment and MALDI-TOF MS for rapid detection of Streptococcus agalactiae colonisation of pregnant women.

Sample preparation was optimized for MALDI-TOF MS directly from selective enrichment broth to detect Streptococcus agalactiae. The method was tested on 100 vaginal samples of pregnant women; positive and negative predictive values were 100 and 91%, respectively. If it indicates positivity, colonisation can be reported 18-24h after sample collection.

Sporadic norovirus infections among hospitalized and non-hospitalized 0-3-year-old infants.

Little is known of the frequency of norovirus infections in sporadic gastroenteritis cases. In a 4-y study, we examined 1152 rota-, adeno- and astrovirus negative stool samples for human noroviruses; 187 (16.2%) proved to be positive. Stool samples were from paediatric patients with acute gastroenteritis. All isolates were verified as sporadic cases: there was no accumulation.