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1.
BMC Microbiol ; 22(1): 30, 2022 01 19.
Article in English | MEDLINE | ID: mdl-35045829

ABSTRACT

BACKGROUND: Recently, a dramatic increase of Klebsiella pneumoniae positive for OXA-48 ß-lactamases was observed first in the hospital setting and later in the long-term care facilities (LTCFs) and community in the Zagreb County, particularly, in urinary isolates. The aim of the study was to analyse the epidemiology and the mechanisms of antibiotic resistance of OXA-48 carbapenemase producing K. pneumoniae strains isolated from urine of non-hospitalized elderly patients. RESULTS: The isolates were classified into two groups: one originated from the LTCFs and the other from the community. Extended-spectrum ß-lactamases (ESBLs) were detected by double disk-synergy (DDST) and combined disk tests in 55% of the isolates (51/92). The ESBL-positive isolates exhibited resistance to expanded-spectrum cephalosporins (ESC) and in majority of cases to gentamicin. LTCFs isolates showed a significantly lower rate of additional ESBLs and consequential resistance to ESC and a lower gentamicin resistance rate compared to the community isolates, similarly to hospital isolates in Zagreb, pointing out to the possible transmission from hospitals.ESBL production was associated with group 1 of CTX-M or SHV-12 ß-lactamases. Ertapenem resistance was transferable from only 12 isolates. blaOXA-48 genes were carried by IncL plasmid in 42 isolates. In addition IncFII and IncFIB were identified in 18 and 2 isolates, respectively. Two new sequence types were reported: ST4870 and ST4781. CONCLUSIONS: This study showed eruptive and extensive diffusion of OXA-48 carbapenemase to LTCFs and community population in Zagreb County, particularly affecting patients with UTIs and urinary catheters. On the basis of susceptibility testing, ß-lactamase production, conjugation experiments, MLST and plasmid characterization it can be concluded that there was horizontal gene transfer between unrelated isolates, responsible for epidemic spread of OXA-48 carbapenemase in the LTCFs and the community The rapid spread of OXA-48 producing K. pneumoniae points out to the shortcomings in the infection control measures.


Subject(s)
Klebsiella Infections/epidemiology , Klebsiella Infections/urine , Klebsiella pneumoniae/enzymology , beta-Lactamases/metabolism , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , Croatia/epidemiology , Drug Resistance, Multiple, Bacterial , Female , Hospitalization , Humans , Klebsiella pneumoniae/drug effects , Male , Microbial Sensitivity Tests , Middle Aged , Multilocus Sequence Typing , beta-Lactamases/genetics
2.
Infection ; 48(6): 853-860, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32462287

ABSTRACT

PURPOSE: Microbial dysbiosis has been found preceding necrotizing enterocolitis (NEC) in preterm infants; thus, we aimed to investigate whether there is evidence that neonates with extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) positive stool cultures are at higher risk for NEC at the NICU. METHODS: We included very preterm inborn infants of ≤ 32 weeks of gestational age being fecal carriers of ESBL-E and compared them with 1:1 matched (gestational age, birth weight, gender and year) controls tested negative for ESBL-E in the stool between 2005 and 2016. An association with NEC was defined as the first detection of ESBL-E before or at the time of definite diagnosis of NEC. RESULTS: During the study period, we diagnosed 217 infants with a total of 270 ESBL-E. We identified ten different species with ESBL-producing Klebsiella oxytoca being the most common one (46%) followed by Klebsiella pneumoniae (19%), and Citrobacter freundii (17%). Ten out of 217 infants had any kind of NEC in the case group compared to two of the controls (p < 0.01), but only four cases with predefined criteria were associated with NEC ≥ stage IIa (1.8 vs. 0.5%, p = 0.089, OR 4.1, CI95% 0.45-36.6). NEC mortality rate was 2/8 (25%). CONCLUSIONS: We observed a threefold increase of ESBL-E in stool surveillance cultures during study time and germs were dominated by ESBL-producing Klebsiella spp. There was no evidence that preterm infants colonized with ESBL-E in the stool were at higher risk for definite NEC.


Subject(s)
Enterobacteriaceae Infections/epidemiology , Enterobacteriaceae/isolation & purification , Enterocolitis, Necrotizing/epidemiology , Feces/microbiology , beta-Lactamases/analysis , Austria/epidemiology , Case-Control Studies , Enterobacteriaceae/metabolism , Enterobacteriaceae Infections/microbiology , Enterocolitis, Necrotizing/microbiology , Female , Humans , Incidence , Infant, Newborn , Infant, Premature , Intensive Care Units, Neonatal , Male , Retrospective Studies , Risk Factors
3.
Chemotherapy ; 64(4): 167-172, 2019.
Article in English | MEDLINE | ID: mdl-31707391

ABSTRACT

INTRODUCTION: Carbapenem resistance in Acinetobacter baumannii can be mediated by carbapenemases of class A, class B metallo-ß-lactamases (MBLs), and class D carbapenem-hydrolyzing oxacillinases (CHDL). The aim of the study was to investigate the antimicrobial susceptibility and ß-lactamase production of carbapenem-resistant A. baumannii isolates (CRAB) from the Children's Hospital Zagreb, Croatia. METHODS: A total of 12 A. baumannii isolates collected between August 2016 and March 2018 were analyzed. Antibiotic susceptibility was determined by the broth microdilution method. The presence of MBLs was explored by combined disk test with EDTA. The presence of carbapenemases of class A, B, and D was explored by PCR. The occurrence of the ISAba1 upstream of the blaOXA-51-like or blaOXA-23-like was determined by PCR mapping. Epidemiological typing was performed by determination of sequence groups (SG). Genotyping was performed by SG determination, rep-PCR, and MLST. RESULTS: All CRAB were resistant to piperacillin/tazobactam, ceftazidime, cefotaxime, ceftriaxone, cefepime, imipenem, meropenem, gentamicin, and ciprofloxacin. Moderate resistance rates were observed for ampicillin/sulbactam (67%) and tigecycline (42%). The isolates were uniformly susceptible to colistin. PCR revealed the presence of genes encoding OXA-24-like CHDL in nine and OXA-23-like CHDL in three isolates. blaOXA-51 genes were preceded by ISAba1. PCR for the common MBLs in Acinetobacter was negative. All isolates belonged to SG 1 corresponding to ICL (International Clonal Lineage) II. Rep-PCR identified four major clones. CONCLUSIONS: The study found OXA-24-like ß-lactamase to be the dominant CHDL among children'sCRAB. The predominant spread of OXA-24-like is in contrast with the recent global dissemination of OXA-23 reported all over the world. In contrast to the previous studies in which emergency of OXA-24-like positive isolates was monoclonal, we found considerable genetic diversity of the isolates.


Subject(s)
Acinetobacter Infections/diagnosis , Acinetobacter baumannii/enzymology , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Acinetobacter Infections/microbiology , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Carbapenems/metabolism , Child , Croatia , Drug Resistance, Multiple, Bacterial/drug effects , Drug Resistance, Multiple, Bacterial/genetics , Genotype , Humans , Hydrolysis , Microbial Sensitivity Tests , Multilocus Sequence Typing , beta-Lactamases/genetics
4.
Curr Microbiol ; 72(5): 606-11, 2016 May.
Article in English | MEDLINE | ID: mdl-26821237

ABSTRACT

Cystic fibrosis (CF) is the most common hereditary lung disease in the Caucasian population, characterized by viscous bronchial secretion, consecutive defective mucociliary clearance, and unavoidable colonization with microorganisms. Besides Pseudomonas aeruginosa, Staphylococcus aureus is the most common bacterial species colonizing the CF respiratory tract. Under antibiotic pressure S. aureus is able to switch to small colony variants (SCV). These small colony variants can invade epithelial cells, overcome antibiotic therapy inside the cells and can be the starting point for extracellular recolonization. The aim of the present study was the isolation and characterization of S. aureus small colony variants from Austrian cystic fibrosis patients. Samples collected from 147 patients were screened for the presence of S. aureus wild-type and small colony variants. Antibiotic susceptibility testing and determination of the small colony variants causing auxotrophism were performed. Wild-type isolates were assigned to corresponding small colony variants with spa typing. In total, 17 different small colony variant isolates and 12 corresponding wild-type isolates were obtained. 13 isolates were determined thymidine auxotroph, 2 isolates were auxotroph for hemin, and none of the tested isolates was auxotroph for both, respectively. The presence of SCVs is directly related to a poor clinical outcome, therefore a monitoring of SCV prevalence is recommended. This study revealed rather low SCV ratios in CF patients compared to other countries.


Subject(s)
Cystic Fibrosis/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification , Adolescent , Adult , Anti-Bacterial Agents/pharmacology , Austria , Child , Child, Preschool , Female , Humans , Infant , Male , Microbial Sensitivity Tests , Middle Aged , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Staphylococcus aureus/growth & development , Young Adult
5.
Anaerobe ; 42: 162-165, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27789247
6.
Antimicrob Agents Chemother ; 59(1): 714-6, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25348541

ABSTRACT

We investigated sinks as possible sources of a prolonged Klebsiella pneumonia carbapenemase (KPC)-producing Klebsiella oxytoca outbreak. Seven carbapenem-resistant K. oxytoca isolates were identified in sink drains in 4 patient rooms and in the medication room. Investigations for resistance genes and genetic relatedness of patient and environmental isolates revealed that all the isolates harbored the blaKPC-2 and blaTEM-1 genes and were genetically indistinguishable. We describe here a clonal outbreak caused by KPC-2-producing K. oxytoca, and handwashing sinks were a possible reservoir.


Subject(s)
Cross Infection/etiology , Equipment Contamination , Klebsiella Infections/etiology , Klebsiella oxytoca , Cross Infection/epidemiology , Cross Infection/microbiology , Disease Outbreaks , Hand Disinfection , Hematology , Hospital Departments , Humans , Klebsiella Infections/epidemiology , Klebsiella Infections/microbiology , Klebsiella oxytoca/enzymology , beta-Lactamases
7.
Int J Med Microbiol ; 305(8): 918-24, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26507866

ABSTRACT

OBJECTIVES: An increase of severe infections caused by Streptococcus dysgalactiae subsp. equisimilis (SDSE) similar to infections caused by Streptococcus pyogenes has been reported over the last years. Little is known about infections with SDSE in Austria. Therefore, we investigated a collection of 113 SDSE invasive and non-invasive isolates from different infection sites and type of infections as well as patients' characteristics. METHODS: The isolates were phenotypically identified and emm typed using the enlarged emm database from the Centers for Disease Control and Prevention. Additionally, 13 antimicrobial agents were tested using EUCAST guidelines and virulence genes were investigated. RESULTS: Severe SDSE infections were most common in elderly men with underlying diseases especially diabetes mellitus. With VitekMS identification of SDSE isolates was successful to the species level only. Emm typing revealed 24 different emm types, one new type and one new subtype. StG485, stG6, stC74a, stG643, and stG480 were the predominant types in this study, stC74a and stG652 in invasive infections and stG643, stC74a and stG485 in non-invasive infections. Resistance was observed to tetracycline (62%), macrolides (13%) with one M phenotype, and clindamycin (12%) presenting 6 constitutive MLS(B) phenotypes and 8 inducible MLS(B) phenotypes. Levofloxacin resistance was detected only in one isolate. All isolates tested for virulence genes were positive for scpA, ska, saga and slo. Superantigenic genes were negative except speG(dys) (positive 17/34; 50%). CONCLUSION: This paper presents the first report of SDSE infections in Austria. Severe SDSE infections were found mainly in elderly men with underlying diseases. SDSE isolates demonstrated substantial emm type diversity without association with infections site or invasiveness. Analysis of virulence genes showed no significant difference between invasive and non-invasive infections.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antigens, Bacterial/analysis , Bacterial Outer Membrane Proteins/analysis , Carrier Proteins/analysis , Genotype , Serogroup , Streptococcal Infections/epidemiology , Streptococcal Infections/microbiology , Streptococcus/classification , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Austria/epidemiology , Child , Child, Preschool , Female , Genetic Variation , Humans , Infant , Male , Microbial Sensitivity Tests , Middle Aged , Prevalence , Risk Factors , Streptococcus/isolation & purification , Young Adult
8.
Eur J Clin Invest ; 45(8): 824-32, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26058473

ABSTRACT

BACKGROUND: Catheter-related bloodstream infections (CRBSIs) are currently detected in patients with clinically suspicion. The aim of our study was to evaluate whether CRBSIs could be anticipated and detected in a subclinical stage by peptide nucleic acid fluorescence in situ hybridization (PNA FISH) using universal hybridization probes or acridine orange leucocyte cytospin (AOLC) tests in haematooncological patients with central venous catheters (CVCs) in situ. MATERIALS AND METHODS: Peptide nucleic acid fluorescence in situ hybridization and AOLC tests using blood samples from one CVC lumen/port chamber in haematooncological patients were continuously performed. These results were compared to those obtained from routinely performed CRBSI diagnostic tests. RESULTS: One hundred and eighty-two patients with 342 catheter periods were investigated. Seventeen CRBSI cases were detected in 6466 CVC days by routine measures resulting in a CRBSI rate of 2.6/1000 catheter days. Two of 17 showed positive PNA FISH tests, and five positive AOLC test results before the diagnosis were established with routine measures. The screening revealed further seven patients with positive universal PNA FISH tests and 10 positive AOLC tests without symptoms indicative for infection and were therefore considered not to have CRBSI. CONCLUSIONS: Sampling of only one CVC lumen/port chamber screening for CRBSI in haematooncological patients seems not to be a useful tool for anticipative diagnosis of CRBSI. Reasons for false-negative results might include origin of CRBSIs from the other CVC lumina not sampled for screening, and false-positive results might origin from catheter colonization without subsequent spread of micro-organisms into the peripheral bloodstream.


Subject(s)
Bacteremia/diagnosis , Catheter-Related Infections/diagnosis , Central Venous Catheters , Fungemia/diagnosis , Hematologic Neoplasms/complications , Acridine Orange , Adult , Aged , Bacteremia/complications , Candidiasis/complications , Candidiasis/diagnosis , Catheter-Related Infections/complications , Cohort Studies , Enterobacteriaceae Infections/complications , Enterobacteriaceae Infections/diagnosis , Escherichia coli Infections/complications , Escherichia coli Infections/diagnosis , False Negative Reactions , Female , Fluorescent Dyes , Fungemia/complications , Gram-Negative Bacterial Infections/complications , Gram-Negative Bacterial Infections/diagnosis , Gram-Positive Bacterial Infections/complications , Gram-Positive Bacterial Infections/diagnosis , Humans , In Situ Hybridization, Fluorescence , Klebsiella Infections/complications , Klebsiella Infections/diagnosis , Male , Middle Aged , Peptide Nucleic Acids , Pseudomonas Infections/complications , Pseudomonas Infections/diagnosis , Staphylococcal Infections/complications , Staphylococcal Infections/diagnosis , Stenotrophomonas maltophilia
9.
Respiration ; 89(5): 390-5, 2015.
Article in English | MEDLINE | ID: mdl-25825042

ABSTRACT

BACKGROUND: Cystic fibrosis (CF) is the most common life-limiting inherited disease in Caucasian populations. While pathological changes can be seen in various organs, morbidity and mortality are mainly related to the respiratory tract, with patients suffering from chronic bronchopulmonary infections with characteristic pathogens including Staphylococcus aureus. OBJECTIVES: To date, there is only very limited data on the genetic and phenotypic characteristics of S. aureus in CF patients. Therefore, in our study, we characterized 58 S. aureus isolates collected from CF patients in Austria by spa typing, DNA microarray profiling, as well as antimicrobial susceptibility testing in order to determine common genomic and antimicrobial resistance features. The tested strain collection exhibited high genomic diversity. RESULTS: The 58 isolates were assigned to 16 clonal complexes and 48 spa types and differed greatly regarding their virulence and resistance gene profiles. The predominant clonal complexes were MLST CC30 (22%), CC15 (16%), CC45 (14%), and CC5 (12%), complexes that are highly prevalent worldwide among S. aureus strains isolated from humans colonized or infected with S. aureus. DNA microarray profiles showed a wide variety of genes encoding antimicrobial resistance and virulence factors such as various leukocidins, haemolysins, enterotoxins, exfoliative toxins, toxic shock syndrome toxin, as well as genes involved in adhesion and immune evasion. CONCLUSIONS: While a large number of strains exhibited resistance to one or several antimicrobial agents, methicillin-resistant S. aureus was found at a low prevalence of 3% (n = 2) only. The two methicillin-resistant S. aureus isolates were assigned to CC152/t355 (SCCmecV) and CC5/t001 (SCCmecI). This is the first study to genetically characterize S. aureus isolates in CF patients in Austria.


Subject(s)
Cystic Fibrosis/microbiology , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity , Bacterial Toxins/genetics , Bacterial Typing Techniques , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Enterotoxins/genetics , Genotype , Humans , Oligonucleotide Array Sequence Analysis , Phenotype , Virulence
10.
Crit Care ; 18(2): R47, 2014 Mar 24.
Article in English | MEDLINE | ID: mdl-24661739

ABSTRACT

INTRODUCTION: Vitamin D plays a key role in immune function. Deficiency may aggravate the incidence and outcome of infectious complications in critically ill patients. We aimed to evaluate the prevalence of vitamin D deficiency and the correlation between serum 25-hydroxyvitamin D (25(OH) D) and hospital mortality, sepsis mortality and blood culture positivity. METHODS: In a single-center retrospective observational study at a tertiary care center in Graz, Austria, 655 surgical and nonsurgical critically ill patients with available 25(OH) D levels hospitalized between September 2008 and May 2010 were included. Cox regression analysis adjusted for age, gender, severity of illness, renal function and inflammatory status was performed. Vitamin D levels were categorized by month-specific tertiles (high, intermediate, low) to reflect seasonal variation of serum 25(OH) D levels. RESULTS: Overall, the majority of patients were vitamin D deficient (<20 ng/ml; 60.2%) or insufficient (≥20 and <30 ng/dl; 26.3%), with normal 25(OH) D levels (>30 ng/ml) present in only 13.6%. The prevalence of vitamin D deficiency and mean 25(OH) D levels was significantly different in winter compared to summer months (P <0.001). Hospital mortality was 20.6% (135 of 655 patients). Adjusted hospital mortality was significantly higher in patients in the low (hazard ratio (HR) 2.05, 95% confidence interval (CI) 1.31 to 3.22) and intermediate (HR 1.92, 95% CI 1.21 to 3.06) compared to the high tertile. Sepsis was identified as cause of death in 20 of 135 deceased patients (14.8%). There was no significant association between 25(OH) D and C-reactive protein (CRP), leukocyte count or procalcitonin levels. In a subgroup analysis (n = 244), blood culture positivity rates did not differ between tertiles (23.1% versus 28.2% versus 17.1%, P = 0.361). CONCLUSIONS: Low 25(OH) D status is significantly associated with mortality in the critically ill. Intervention studies are needed to investigate the effect of vitamin D substitution on mortality and sepsis rates in this population.


Subject(s)
Critical Illness , Hospital Mortality/trends , Seasons , Sepsis/blood , Vitamin D Deficiency/blood , Vitamin D/analogs & derivatives , Adult , Aged , Aged, 80 and over , Biomarkers/blood , Critical Illness/mortality , Female , Humans , Male , Middle Aged , Retrospective Studies , Sepsis/diagnosis , Sepsis/mortality , Vitamin D/blood , Vitamin D Deficiency/diagnosis , Vitamin D Deficiency/mortality
11.
Food Microbiol ; 44: 41-6, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25084643

ABSTRACT

INTRODUCTION: Resistant bacteria are a well-known public health problem. This study was conducted to investigate the prevalence and genetic characteristics of extended spectrum ß-lactamase (ESBL) producing enterobacteria, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) in mixed minced meat from pork and beef. METHODS: One hundred samples of mixed minced meat were collected from supermarkets (n = 70) and local butcher shops (n = 30) in the city of Graz (Austria). After enrichment and inoculation on selective media, bacteria were identified with MALDI-TOF MS or Vitek2 systems, tested for antibiotic resistance and further characterized with PCR and sequencing. RESULTS: In 20 of the 100 meat samples 24 ESBL positive Escherichia coli isolates were found. The most common ESBL among the isolates was CTX-M-1. Other detected bla genes contained CTX-M-14, CTX-M-32, SHV-12 and TEM-52 types. Nine samples were tested positive for MRSA and spa-typed. Detected spa-types were hospital-acquired t3928, as well as livestock-associated t011, t034 and t2241. No VRE were found. CONCLUSION: A contamination of meat with ESBL-producing E. coli and MRSA was confirmed in this study. The large diversity of ESBL producing E. coli could indicate a growing dissemination of ESBL genes in E. coli found in meat products from porcine and bovine origin.


Subject(s)
Bacteria/drug effects , Bacteria/isolation & purification , Drug Resistance, Multiple, Bacterial , Food Contamination/analysis , Meat/microbiology , Animals , Anti-Bacterial Agents/pharmacology , Austria , Bacteria/classification , Bacteria/genetics , Cattle , Poultry , Swine
12.
Pathogens ; 13(5)2024 May 15.
Article in English | MEDLINE | ID: mdl-38787264

ABSTRACT

Multidrug-resistant (MDR) bacteria of the utmost importance are extended-spectrum ß-lactamase (ESBL) and carbapenemase-producing Enterobacterales (CRE), carbapenem-resistant Acinetobacter baumannii (CRAB), carbapenem-resistant Pseudomonas aeruginosa (CRPA), methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus spp. (VRE). In this study, an evaluation of MDR bacteria in surgical intensive care units in a tertiary referral hospital was conducted. The study aimed to characterize ß-lactamases and other resistance traits of Gram-negative bacteria isolated in surgical intensive care units (ICUs). Disk diffusion and the broth dilution method were used for antibiotic susceptibility testing, whereas ESBL screening was performed through a double disk synergy test and an inhibitor-based test with clavulanic acid. A total of 119 MDR bacterial isolates were analysed. ESBL production was observed in half of the Proteus mirabilis, 90% of the Klebsiella pneumoniae and all of the Enterobacter cloacae and Escherichia coli isolates. OXA-48 carbapenemase, carried by the L plasmid, was detected in 34 K. pneumoniae and one E. coli and Enterobacter cloacae complex isolates, whereas NDM occurred sporadically and was identified in three K. pneumoniae isolates. OXA-48 positive isolates coharboured ESBLs belonging to the CTX-M family in all but one isolate. OXA-23 carbapenemase was confirmed in all A. baumannii isolates. The findings of this study provide valuable insight of resistance determinants of Enterobacterales and A. baumannii which will enhance surveillance and intervention strategies that are necessary to curb the ever-growing carbapenem resistance rates.

14.
J Clin Microbiol ; 51(12): 3944-9, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24025914

ABSTRACT

Successful multidrug-resistant clones are increasing in prevalence globally, which makes the ability to identify these clones urgent. However, adequate, easy-to-perform, and reproducible typing methods are lacking. We investigated whether DiversiLab (DL), an automated repetitive-sequence-based PCR bacterial typing system (bioMérieux), is suitable for comparing isolates analyzed at different geographic centers. A total of 39 Escherichia coli and 39 Klebsiella species isolates previously typed by the coordinating center were analyzed. Pulsed-field gel electrophoresis (PFGE) confirmed the presence of one cluster of 6 isolates, three clusters of 3 isolates, and three clusters of 2 isolates for each set of isolates. DL analysis was performed in 11 centers in six different countries using the same protocol. The DL profiles of 425 E. coli and 422 Klebsiella spp. were obtained. The DL system showed a lower discriminatory power for E. coli than did PFGE. The local DL data showed a low concordance, as indicated by the adjusted Rand and Wallace coefficients (0.132 to 0.740 and 0.070 to 1.0 [E. coli] and 0.091 to 0.864 and 0.056 to 1.0 [Klebsiella spp.], respectively). The central analysis showed a significantly improved concordance (0.473 to 1.0 and 0.290 to 1.0 [E. coli] and 0.513 to 0.965 and 0.425 to 1.0 [Klebsiella spp.], respectively). The misclassifications of profiles for individual isolates were mainly due to inconsistent amplification, which was most likely due to variations in the quality and amounts of the isolated DNA used for amplification. Despite local variations, the DL system has the potential to indicate the occurrence of clonal outbreaks in an international setting, provided there is strict adherence to standardized, reproducible DNA isolation methods and analysis protocols, all supported by a central database for profile comparisons.


Subject(s)
Escherichia coli/classification , Klebsiella/classification , Molecular Typing/methods , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Escherichia coli/genetics , Humans , International Cooperation , Klebsiella/genetics , Polymerase Chain Reaction/methods , Reproducibility of Results
15.
Eur J Clin Invest ; 43(9): 964-9, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23848936

ABSTRACT

BACKGROUND: Catheter-related bloodstream infections (CRBSIs) are currently detected with a reactive diagnostic policy, that is, application of tests to patients with clinically suspected CRBSI. The aim of our study was to evaluate whether CRBSIs could be anticipated in an earlier stage by microbiological screening using peptide nucleic acid fluorescence in situ hybridization (PNA FISH) with universal hybridization probes or acridine-orange leucocyte cytospin (AOLC) tests in haemodialysis and haematological patients with CVCs in situ compared with routine test. MATERIALS AND METHODS: Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) and AOLC tests using blood samples from both CVC lines in patients undergoing haemodialysis were performed three times a week and from one CVC line in haematological patients were performed daily. Results were compared with those obtained from routinely performed CRBSI diagnostic tests. RESULTS: One hundred fifteen patients with 139 catheter periods were investigated. The mean observation time per catheter period was 25 days (IQR 13.5-43.5), resulting in 5615 CVC days with a total of 4839 tested blood samples. Five CRBSI cases were detected by routine measures resulting in a CRBSI rate of 0.9/1000 catheter days. Four of five CRBSIs could be anticipated by positive PNA FISH and AOLC tests 2-8 days before the diagnosis was established with routine measures. CONCLUSIONS: The proactive anticipative strategy using microscopic examination of CVC blood samples to anticipate CRBSI in an earlier stage compared with routine measures is a new diagnostic approach in patients with CVCs and a high risk of developing CRBSI.


Subject(s)
Catheter-Related Infections/diagnosis , Central Venous Catheters , Acridine Orange , Adult , Aged , Early Diagnosis , Fluorescent Dyes , Hematologic Neoplasms/therapy , Hematopoietic Stem Cell Transplantation , Humans , In Situ Hybridization, Fluorescence/methods , Microbiological Techniques , Middle Aged , Peptide Nucleic Acids/metabolism , Prospective Studies , Renal Dialysis , Young Adult
16.
Pediatr Crit Care Med ; 14(2): 157-63, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23254982

ABSTRACT

OBJECTIVE: Fecal carriage of extended-spectrum ß-lactamase-producing enterobacteriaceae may contribute to the spread of extended-spectrum ß-lactamase-producing enterobacteriaceae into the community. The objective of this study was to assess the duration of fecal carriage after discharge and the occurrence of intrafamilial transmission. DESIGN: Case series. SETTING: Quaternary care children's hospital. PATIENTS: Patients colonized with extended-spectrum ß-lactamase-producing enterobacteriaceae at the neonatal ICU and the respective household members. INTERVENTIONS: Screening for intestinal extended-spectrum ß-lactamase-producing enterobacteriaceae colonization was done at 1, 2, 4, 6, 9, and 12 months after discharge. Genetic relatedness of isolated extended-spectrum ß-lactamase-producing enterobacteriaceae strains was determined using automated rep-PCR. RESULTS: Twenty-five neonates (case-patients) colonized with extended-spectrum ß-lactamase-producing enterobacteriaceae (one extended-spectrum ß-lactamase-Escherichia coli; six extended-spectrum ß-lactamase-Klebsiella pneumoniae; 11 extended-spectrum ß-lactamase-Klebsiella oxytoca; and seven extended-spectrum ß-lactamase-Serratia marcescens) were included. Duration of fecal carriage was longer (up to 1 yr) in case-patients colonized with Klebsiella species than in case-patients colonized with Serratia marcescens (<4 months). During follow-up, strains and species of extended-spectrum ß-lactamase-producing enterobacteriaceae different from the primary strain were found in four and three case-patients, respectively. In nine of 49 (18.4%) included household members, extended-spectrum ß-lactamase-producing enterobacteriaceae were found during the follow-up period. In two of nine colonized household members, the isolated extended-spectrum ß-lactamase-producing enterobacteriaceae was identical to the primary strains of the respective case-patients. CONCLUSIONS: After intestinal colonization with extended-spectrum ß-lactamase-producing enterobacteriaceae at the neonatal ICU, infants potentially remain carriers during the first year after discharge. Intrafamilial spread has been proven.


Subject(s)
Carrier State/microbiology , Enterobacteriaceae Infections/transmission , Enterobacteriaceae/metabolism , Feces/microbiology , beta-Lactamases/biosynthesis , Cross Infection/microbiology , Enterobacteriaceae/genetics , Enterobacteriaceae Infections/microbiology , Escherichia coli/genetics , Escherichia coli/isolation & purification , Family , Female , Humans , Infant , Infant, Newborn , Intensive Care Units, Neonatal , Klebsiella oxytoca/genetics , Klebsiella oxytoca/isolation & purification , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Male , Serratia marcescens/genetics , Serratia marcescens/isolation & purification , Time Factors
17.
J Infect Chemother ; 19(6): 1214-7, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23744026

ABSTRACT

We report the emergence of OXA-48 carbapenemase-producing Escherichia coli in Austria causing ventilator-associated pneumonia in a traveler returning from Egypt. Depending on resistance testing, quinolones may remain a therapeutic option for infections caused by these multiple resistant pathogens, as this class of drugs has a favorable safety and tolerability profile when compared to the alternatives. In this patient, however, the clinical course was dramatically complicated by the development of ciprofloxacin-associated rhabdomyolysis.


Subject(s)
Ciprofloxacin/adverse effects , Escherichia coli Infections/etiology , Escherichia coli/isolation & purification , Pneumonia, Ventilator-Associated/microbiology , Rhabdomyolysis/chemically induced , Rhabdomyolysis/microbiology , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/therapeutic use , Ciprofloxacin/therapeutic use , Escherichia coli Infections/microbiology , Escherichia coli Proteins , Humans , Male , Middle Aged , beta-Lactamases
18.
Pathogens ; 12(11)2023 Nov 02.
Article in English | MEDLINE | ID: mdl-38003773

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) is a universal threat. Once being well established in the healthcare setting, MRSA has undergone various epidemiological changes. This includes the emergence of more aggressive community-acquired MRSA (CA-MRSA) and the occurrence of MRSA which have their origin in animal breeding, called livestock-associated MRSA (LA-MRSA). Emergence of new clones as well as changes in the occurrence of some clonal lineages also describes the fluctuating dynamic within the MRSA family. There is paucity of data describing the possible impact of the COVID-19 pandemic on the MRSA dynamics. The aim of the study was the analysis of MRSA isolates in a three-year time period, including the pre-COVID-19 years 2018 and 2019 and the first year of the pandemic 2020. The analysis includes prevalence determination, antibiotic susceptibility testing, spa typing, and detection of genes encoding the PVL toxin. The MRSA rate remained constant throughout the study period. In terms of a dynamic within the MRSA family, only a few significant changes could be observed, but all except one occurred before the start of the COVID-19 pandemic. In summary, there was no significant impact of the COVID-19 pandemic on MRSA in Austria.

19.
Anat Sci Int ; 98(1): 151-154, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36471134

ABSTRACT

Post-mortem specimens used for anatomy teaching are commonly embalmed using compositions of chemicals, with the objective to maintain tissue quality and to avoid putrefaction. Monitoring for bacterial or fungal contamination is becoming increasingly important especially when measures are taken to minimize exposure by chemicals such as formaldehyde. In this case, random swabs were taken from six corpses embalmed with ethanol-glycerin and Thiel embalming. Cultures and MALDI-TOF analyses yielded four cases of Clostridium perfringens contamination. C. perfringens is of special interest as a human pathogen. A potential source was identified in the containers filled with the moistening solution. Cross contamination with Clostridium species has likely occurred between corpses sharing the moistening solution and soaking the cover linen directly within the containers. To minimize any risk for those exposed, the moistening solutions were discarded and all equipment thoroughly disinfected. The specimens had to be cremated as they formed a potential source of Clostridium spores. Deviating from previous routines it was formalized that the cover linen must not be submerged in the moistening contains rather than moistening the specimens directly with dedicated vessels. Follow-up analyses yielded no further contamination with C. perfringens.


Subject(s)
Clostridium perfringens , Incidental Findings , Humans , Formaldehyde , Ethanol , Cadaver
20.
Pathogens ; 12(1)2023 Jan 10.
Article in English | MEDLINE | ID: mdl-36678465

ABSTRACT

During November to December 2020, a high rate of COVID-19-associated pneumonia with bacterial superinfections due to multidrug-resistant (MDR) pathogens was recorded in a COVID-19 hospital in Zagreb. This study analyzed the causative agents of bacterial superinfections among patients with serious forms of COVID-19. In total, 118 patients were hospitalized in the intensive care unit (ICU) of the COVID-19 hospital. Forty-six out of 118 patients (39%) developed serious bacterial infection (VAP or BSI or both) during their stay in ICU. The total mortality rate was 83/118 (70%). The mortality rate due to bacterial infection or a combination of ARDS with bacterial superinfection was 33% (40/118). Six patients had MDR organisms and 34 had XDR (extensively drug-resistant). The dominant species was Acinetobacter baumannii with all isolates (34) being carbapenem-resistant (CRAB) and positive for carbapenem-hydrolyzing oxacillinases (CHDL). One Escherichia coli causing pneumonia harboured the blaCTX-M-15 gene. It appears that the dominant resistance determinants of causative agents depend on the local epidemiology in the particular COVID center. Acinetobacter baumannii seems to easily spread in overcrowded ICUs. Croatia belongs to the 15 countries in the world with the highest mortality rate among COVID-19 patients, which could be in part attributable to the high prevalence of bacterial infections in local ICUs.

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