[Resistance of bacterial pathogens isolated from the lower respiratory tract and their clonality in intensive care patients in a post-COVID-19 period].

OBJECTIVES: The period of the COVID-19 pandemic had a significant impact on the healthcare system, including its effect on compliance with the established procedures of a rational antibiotic policy, especially in the context of nosocomial pneumonia, where it was very difficult to distinguish a possible bacterial superinfection from a severe inflammatory reaction caused by the SARS-CoV-2 virus. The aim of the present study was to analyze the antimicrobial resistance of bacterial pathogens isolated from the lower respiratory tract and their clonality in intensive care patients in 2022 and to compare it with the previous COVID-19 period. MATERIAL AND METHODS: Bacterial strains isolated from the lower respiratory tract (LRT) of patients hospitalized at the Department of Anaesthesiology, Resuscitation and Intensive Care, Olomouc University Hospital (DARIC) over a three-year period (January 1, 2020 - December 31, 2022) were included in the study. The susceptibility to antibiotics was determined by the standard microdilution method according to the EUCAST criteria, and selected isolates were compared using pulsed-field gel electrophoresis (PFGE). RESULTS: The resistance of the most common bacterial pathogens isolated from the LRT of patients hospitalized at DARIC did not change significantly during the COVID-19 (2020-2021) and post-COVID-19 (2022) periods, with the exception of Serratia marcescens and Enterococcus faecium species. These two showed an increase in the number of strains during the COVID-19 pandemic, as well as a significant increase in the proportion of resistant strains. In the case of Serratia marcescens, there was a subsequent decrease in the number of isolates and their resistance in 2022. For Enterococcus faecium, the total number of isolates also decreased significantly, but the frequency of vancomycin-resistant isolates (VRE) continued to increase. During the COVID-19 pandemic, increased VRE detection can be linked to proven clonal spread, but significant clonality was no longer confirmed in 2022. Comparison of similarity by PFGE in other bacterial species also did not reveal significant horizontal transmission between patients in the post-COVID-19 period, as most isolates (85%) showed a unique restriction profile. CONCLUSIONS: The results indicate that the frequency and antimicrobial resistance of the majority of the most common bacterial pathogens from the LRT of patients hospitalized at DARIC in the post-pandemic period remain comparable to the time before and during the COVID-19 pandemic outbreak. An exception is Enterococcus faecium, which showed an increase in vancomycin resistance in both the COVID-19 and the post-COVID-19 periods.

[Impact of the COVID-19 pandemic on antimicrobial resistance].

OBJECTIVES: The COVID-19 pandemic has had a major impact on the healthcare system, which has been forced to manage large numbers of patients, including those with respiratory insufficiency and in need of oxygen therapy. Due to concerns about bacterial co-infection, antibiotic therapy was administered to many patients. The aim of the present study was to compare antimicrobial resistance in intensive care patients in the pre-pandemic and pandemic periods. MATERIAL AND METHODS: Patients hospitalized at the Department of Anesthesiology, Resuscitation and Intensive Care Medicine of the University Hospital Olomouc in the pre-COVID-19 period (2018-2019) and during the pandemic (2020-2021) were enrolled in the study. Clinical samples from the lower respiratory tract were routinely collected twice a week, with one strain of a given species first isolated from each patient being included in the study. RESULTS: While several bacterial species (Escherichia coli, Proteus mirabilis and Haemophilus influenzae) were found to occur less frequently, an increased occurrence was documented for Enterococcus faecium, Serratia marcescens and Klebsiella variicola. Overall, however, it can be concluded that there was no major change in the frequency of bacterial pathogens isolated from the lower respiratory tract during the COVID-19 period. Similarly, with only a few exceptions, antimicrobial resistance did not change significantly. More significant increases in resistance to piperacillin/tazobactam, cefotaxime, ciprofloxacin and gentamicin have been demonstrated for Serratia marcescens. However, a decrease in the resistance of Pseudomonas aeruginosa and Burkholderia cepacia complex to meropenem was also observed. CONCLUSIONS: There was no significant change in the frequency of bacterial pathogens and their resistance to antibiotics during the COVID-19 pandemic. However, there was an increase or decrease in the percentage of some species and in their resistance.

[Enterococcal infections and their treatment options].

AIM: The study aimed to characterize enterococcal infections at the University Hospital Olomouc and to define antibiotic treatment options. MATERIAL AND METHODS: The data was obtained from the ENVIS LIMS laboratory information system. Between 1 January 2015 and 31 December 2019, clinically relevant enterococci in the hospital and their resistance to antibiotics were retrospectively evaluated. Until mid-2016, criteria defined by Facklam and Collins and biochemical properties determined with the Encoccus test were used for identification. Subsequently, all enterococci were identified using the MALDI-TOF MS system. The susceptibility to antibiotics was determined using a standard microdilution method according to the EUCAST criteria. RESULTS: A total of 8 239 clinically relevant enterococci were isolated over the 5-year period. The most frequently isolated species were Enterococcus faecalis and Enterococcus faecium, which accounted for more than 90% in the period 2017-2019. Enterococci were most frequently isolated from urine (35 %), surgical wounds (17 %) and urethral/vaginal swabs (17 %). Clinically relevant enterococci were most commonly isolated from patients with oncological diagnoses (22%), those with urinary and genital diseases (15%) and respiratory diseases (9%). Enterococcus faecalis strains showed very low resistance to the antibiotics tested. Enterococcus faecium was shown to have 24 % proportion of vancomycin-resistant strains (VRE). CONCLUSION: Primary antibiotics suitable for treating infections with the etiological role of Enterococcus faecalis include aminopenicillins, in case of severe infections in combination with aminoglycosides, in particular gentamicin. For Enterococcus faecium strains, glycopeptides must be chosen. To treat VRE, linezolid or tigecycline are indicated.

[Bacteremia pathogens in the University Hospital Olomouc].

OBJECTIVES: To provide an overview of the most common bacterial species isolated from blood cultures in the University Hospital Olomouc in the years 2015-2019 and their antibiotic resistance patterns. MATERIAL AND METHODS: The data were obtained from the laboratory information management system ENVIS LIMS. The results were analyzed retrospectively for the period from January 1, 2015 to December 31, 2019. Among positive blood cultures, the prevalence of bacterial species was assessed and the most frequent species were evaluated for resistance to selected antibiotics. Each sample was processed using standard microbiology methods with the MALDI-TOF MS system. Susceptibility to antibiotics was tested with the microdilution method according to the EUCAST recommendations. RESULTS: Over the study period, a total of 3 400 isolates from blood cultures were included. Coagulase-negative staphylococci were the most prevalent (37 %), followed by Escherichia coli (16 %), Klebsiella pneumoniae (9 %), Staphylococcus aureus (7 %), Streptococcus spp. (5 %), Pseudomonas aeruginosa (4 %), Enterobacter cloacae (2 %), Enterococcus faecalis (2 %) and Enterococcus faecium (2 %). Resistance of E. coli to 3rd generation cephalosporins, piperacillin/tazobactam, gentamicin and fluoroquinolones ranged from 7 % to 33 %. A high percentage of Klebsiella pneumoniae strains (33 %-65 %) was resistant to the above antibiotics. The prevalence of methicillin-resistant Staphylococcus aureus reached 3-7 %. Pseudomonas aeruginosa exhibited resistance to piperacillin/tazobactam, ceftazidime, cefepime, meropenem, ciprofloxacin and gentamicin ranging from 3 % to 46 %. CONCLUSION: Knowing the prevalence of bacterial species from blood cultures and their antimicrobial resistance patterns is important for empirical antibiotic therapy in case of an existing infection. It is advisable to conduct such epidemiological studies as part of antibiotic stewardship.

[Antibiotic therapy of hospital-acquired pneumonia and its pharmacoeconomics].

BACKGROUND: Important hospital-acquired infections include pneumonia, mainly because of the increasing resistance of bacterial pathogens to antimicrobials and the associated potential failure of antibiotic therapy. The present study aimed at determining the most frequent etiological agents of hospital-acquired pneumonia (HAP) and assessing the relationship between 30-day mortality and adequacy of antibiotic therapy. Based on the obtained information, optimal patterns of antibiotic therapy were to be defined, including a pharmacoeconomic perspective. METHODS: In patients with clinically confirmed HAP, bacterial etiological agents were identified, their susceptibility to antimicrobials was determined and statistical methods were used to assess the relationship between adequacy of antibiotic therapy and 30-day mortality. RESULTS: The study comprised 68 patients with clinically confirmed HAP. The most common etiological agents were strains of Pseudomonas aeruginosa (30.8 %), Klebsiella pneumoniae (23.1 %) and Burkholderia cepacia complex (15.4 %). Gram-negative bacteria accounted for 86.5 % of all bacterial pathogens. The overall mortality reached 42.5 %. In the subgroup of patients with inadequate antibiotic therapy, 30-day mortality was significantly higher (83.3 %) than in the subgroup with adequate therapy (30.0 %; p = 0.002). The risk for 30-day mortality was 2.78 times higher in case of inadequate antibiotic therapy (95%CI: 1.52-5.07). The proportion of Pseudomonas aeruginosa strains was significantly higher in the subgroup of patients with inadequate antibiotic therapy than in those with adequate therapy (67 % vs. 27 %; p = 0.032). CONCLUSION: Results of the present study suggest a significant relationship between mortality of patients with HAP and ineffective antibiotic therapy due to resistance of the bacterial pathogen. Thus, it is clear that initial antibiotic therapy must be based on qualified assumption of sufficient activity against the most common bacterial pathogens and results of surveillance of bacterial resistance in the relevant epidemiological unit. At the same time, however, it must be stressed that it is impossible to cover all potential variants of the etiological agents and their resistance phenotypes.

[Bacterial pathogens causing hospital-acquired pneumonia - a multicenter study in the Czech Republic]. / Bakteriální puvodci nozokomiálních pneumonií - multicentrická stidie v Ceské republice.

INTRODUCTION: The objectives were to assess the prevalence of etiologic agents of hospital-acquired pneumonia (HAP) in patients staying in four big hospitals in the Czech Republic and requiring artificial ventilation. The resistance of the isolated pathogens to antibiotics was determined and initial antibiotic therapy was discussed. METHODS: Included in the study were 155 patients with HAP staying from May 1, 2013 to January 31, 2014 in the Departments of Anesthesiology and Critical Care in the following four centers: Thomayer Hospital Prague, University Hospital Brno, University Hospital Hradec Králové and University Hospital Olomouc. From these patients, endotracheal secretion samples were processed using standard microbiology methods. In identified bacterial strains, susceptibility to antibiotics was tested with the microdilution method according to the EUCAST recommendations. Production of ESBL and AmpC beta-lactamases was detected by disk diffusion tests specific for the particular enzymes. ESBL- and AmpC-positive isolates were subjected to basic genetic analysis. RESULTS: Over the study period, a total of 266 isolates were obtained from 140 patients, with 15 patients having negative culture result. Late-onset pneumonia was present in 72 %. Gram-negative bacteria were most prevalent (81 %), namely Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli. Phenotypic tests for production of broad spectrum beta-lactamases were positive in 37 % of Enterobacteriaceae. Genes for CTX-M, SHV, TEM beta-lactamases or CIT and DHA types of AmpC enzymes were detected. No carbapenemase-producing bacteria, methicillin-resistant Staphylococcus aureus or vancomycin-resistant enterococci were detected. CONCLUSION: The study showed that HAP in the Czech Republic was mostly of Gram-negative etiology. Variable antibiotic susceptibility in the two most frequent etiological agents (Pseudomonas aeruginosa and Klebsiella pneumoniae) resulted in severe therapeutic difficulties. A total of 49 % of patients received inadequate therapy. This fact suggests the impact of antibiotic resistance on intensive care patients´ survival or death. Our study confirmed that one in three patients dies because of HAP.

Bacterial Community- and Hospital-Acquired Pneumonia in Patients with Critical COVID-19-A Prospective Monocentric Cohort Study.

The impact of bacterial pneumonia on patients with COVID-19 infection remains unclear. This prospective observational monocentric cohort study aims to determine the incidence of bacterial community- and hospital-acquired pneumonia (CAP and HAP) and its effect on mortality in critically ill COVID-19 patients admitted to the intensive care unit (ICU) at University Hospital Olomouc between 1 November 2020 and 31 December 2022. The secondary objectives of this study include identifying the bacterial etiology of CAP and HAP and exploring the capabilities of diagnostic tools, with a focus on inflammatory biomarkers. Data were collected from the electronic information hospital system, encompassing biomarkers, microbiological findings, and daily visit records, and subsequently evaluated by ICU physicians and clinical microbiologists. Out of 171 patients suffering from critical COVID-19, 46 (27%) had CAP, while 78 (46%) developed HAP. Critically ill COVID-19 patients who experienced bacterial CAP and HAP exhibited higher mortality compared to COVID-19 patients without any bacterial infection, with rates of 38% and 56% versus 11%, respectively. In CAP, the most frequent causative agents were chlamydophila and mycoplasma; Enterobacterales, which were multidrug-resistant in 71% of cases; Gram-negative non-fermenting rods; and Staphylococcus aureus. Notably, no strains of Streptococcus pneumoniae were detected, and only a single strain each of Haemophilus influenzae and Moraxella catarrhalis was isolated. The most frequent etiologic agents causing HAP were Enterobacterales and Gram-negative non-fermenting rods. Based on the presented results, commonly used biochemical markers demonstrated poor predictive and diagnostic accuracy. To confirm the diagnosis of bacterial CAP in our patient cohort, it was necessary to assess the initial values of inflammatory markers (particularly procalcitonin), consider clinical signs indicative of bacterial infection, and/or rely on positive microbiological findings. For HAP diagnostics, it was appropriate to conduct regular detailed clinical examinations (with a focus on evaluating respiratory functions) and closely monitor the dynamics of inflammatory markers (preferably Interleukin-6).

[Influence of carbapenem selection pressure on bacterial resistance]. / Vliv selekcního tlaku karbapenemu na bakteriální rezistenci.

As a result of high resistance of bacterial pathogens to broad-spectrum penicillins and cephalosporins, carbapenems have been increa-singly used recently. The presented study aimed at analyzing the association between carbapenem consumption and resistance of selected Gramnegative pathogens to meropenem. Using linear regression analysis, a statistically significant association was found between carbapenem consumption and resistance of Pseudomonas aeruginosa.

[Effect of colistin consumption and prevalence of colistin-resistant bacteria]. / Vliv spotreby kolistinu na vyskyt kolistin-rezistentnich bakterii.

OBJECTIVE: Recently, there has been a renaissance of the use of the antibiotic colistin resulting from increasing resistance of bacterial pathogens, particularly in intensive care patients. The study aimed at assessing the impact of colistin consumption on the prevalence of colistin-resistant bacteria in the University Hospital Olomouc (UHO). METHODS: A laboratory database was retrospectively searched to identify all clinically significant colistin-resistant bacterial strains isolated between 2007 and 2011. These data were compared with colistin consumption over the same period and the results were statistically processed. RESULTS: Over the study period, a total of 6 338 clinically significant colistin-resistant strains were detected in the UHO (Acinetobacter spp., Burkholderia cepacia complex, Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp., Morganella morganii, Proteus spp., Providencia spp., Pseudomonas spp., Serratia marcesces and Stenotrophomonas maltophilia). Over the same period, the consumption of colistin increased nearly 10-fold. With the increasing colistin consumption, the numbers of colistin-resistant strains of Pseudomonas aeruginosa and Acinetobacter spp. decreased over that period of time. By contrast, there was an increase in the rates of naturally Burkholderia cepacia complex strains naturally resistant to colistin. An alarming finding is that the prevalence of colistin-resistant strains of Klebsiella pneumoniae increased in the last years of the study period, especially in intensive care patients. CONCLUSIONS: In the UHO, higher consumption of colistin was accompanied by increased numbers of colistin-resistant strains. There was a marked increase of Burkholderia cepacia complex strains and, recently, a statistically insignificant but alarming increase in colistin-resistant Klebsiella pneumoniae strains.

Bacterial Resistance to Antibiotics and Clonal Spread in COVID-19-Positive Patients on a Tertiary Hospital Intensive Care Unit, Czech Republic.

This observational retrospective study aimed to analyze whether/how the spectrum of bacterial pathogens and their resistance to antibiotics changed during the worst part of the COVID-19 pandemic (1 November 2020 to 30 April 2021) among intensive care patients in University Hospital Olomouc, Czech Republic, as compared with the pre-pandemic period (1 November 2018 to 30 April 2019). A total of 789 clinically important bacterial isolates from 189 patients were cultured during the pre-COVID-19 period. The most frequent etiologic agents causing nosocomial infections were strains of Klebsiella pneumoniae (17%), Pseudomonas aeruginosa (11%), Escherichia coli (10%), coagulase-negative staphylococci (9%), Burkholderia multivorans (8%), Enterococcus faecium (6%), Enterococcus faecalis (5%), Proteus mirabilis (5%) and Staphylococcus aureus (5%). Over the comparable COVID-19 period, a total of 1500 bacterial isolates from 372 SARS-CoV-2-positive patients were assessed. While the percentage of etiological agents causing nosocomial infections increased in Enterococcus faecium (from 6% to 19%, p < 0.0001), Klebsiella variicola (from 1% to 6%, p = 0.0004) and Serratia marcescens (from 1% to 8%, p < 0.0001), there were significant decreases in Escherichia coli (from 10% to 3%, p < 0.0001), Proteus mirabilis (from 5% to 2%, p = 0.004) and Staphylococcus aureus (from 5% to 2%, p = 0.004). The study demonstrated that the changes in bacterial resistance to antibiotics are ambiguous. An increase in the frequency of ESBL-positive strains of some species (Serratia marcescens and Enterobacter cloacae) was confirmed; on the other hand, resistance decreased (Escherichia coli, Acinetobacter baumannii) or the proportion of resistant strains remained unchanged over both periods (Klebsiella pneumoniae, Enterococcus faecium). Changes in pathogen distribution and resistance were caused partly due to antibiotic selection pressure (cefotaxime consumption increased significantly in the COVID-19 period), but mainly due to clonal spread of identical bacterial isolates from patient to patient, which was confirmed by the pulse field gel electrophoresis methodology. In addition to the above shown results, the importance of infection prevention and control in healthcare facilities is discussed, not only for dealing with SARS-CoV-2 but also for limiting the spread of bacteria.

Phenotypic detection of broad-spectrum beta-lactamases in microbiological practice.

BACKGROUND: Enterobacteriaceae producing ESBL and AmpC enzymes can be associated with failure of antibiotic therapy and related morbidity and mortality. Their routine detection in microbiology laboratories is still a problem. The aim of this study was to compare the sensitivity of selected phenotypic methods. MATERIAL/METHODS: A total of 106 strains of the Enterobacteriaceae family were tested, in which molecular biology methods confirmed the presence of genes encoding ESBL or AmpC. In ESBL-positive strains, the sensitivity of the ESBL Etest (AB Biodisk) and a modified double-disk synergy test (DDST) were evaluated. AmpC strains were tested by a modified AmpC disk method using 3-aminophenylboronic acid. For simultaneous detection of ESBL and AmpC, the microdilution method with a modified set of antimicrobial agents was used. RESULTS: The sensitivity of the ESBL Etest was 95%; the modified DDST yielded 100% sensitivity for ESBL producers and the AmpC test correctly detected 95% of AmpC-positive strains. The sensitivity of the modified microdilution method was 87% and 95% for ESBL and AmpC beta lactamases, respectively. CONCLUSIONS: The detection of ESBL and AmpC beta lactamases should be based on specific phenotypic methods such as the modified DDST, ESBL Etest, AmpC disk test and the modified microdilution method.

Implementation of Antibiotic Stewardship in a University Hospital Setting.

The article describes activities of an antibiotic center at a university hospital in the Czech Republic and presents the results of antibiotic stewardship program implementation over a period of 10 years. It provides data on the development of resistance of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus to selected antibiotic agents as well as consumption data for various antibiotic classes. The genetic basis of resistance to beta-lactam antibiotics and its clonal spread were also assessed. The study showed significant correlations between aminoglycoside consumption and resistance of Escherichia coli and Klebsiella pneumoniae to gentamicin (r = 0.712, r = 0.869), fluoroquinolone consumption and resistance of Klebsiella pneumoniae to ciprofloxacin (r = 0.896), aminoglycoside consumption and resistance of Pseudomonas aeruginosa to amikacin (r = 0.716), as well as carbapenem consumption and resistance of Pseudomonas aeruginosa to meropenem (r = 0.855). Genotyping of ESBL- positive isolates of Klebsiella pneumoniae and Escherichia coli showed a predominance of CTX-M-type; in AmpC-positive strains, DHA, EBC and CIT enzymes prevailed. Of 19 meropenem-resistant strains of Klebsiella pneumoniae, two were identified as NDM-positive. Clonal spread of these strains was not detected. The results suggest that comprehensive antibiotic stewardship implementation in a healthcare facility may help to maintain the effectiveness of antibiotics against bacterial pathogens. Particularly beneficial is the work of clinical microbiologists who, among other things, approve administration of antibiotics to patients with bacterial infections and directly participate in their antibiotic therapy.

Identification of novel OXA-134-like ß-lactamases in Acinetobacter lwoffii and Acinetobacter schindleri isolated from chicken litter.

BACKGROUND: Various food-producing animals have been recognized in recent years as a potential reservoir for the spread of antibiotic resistant bacteria that may pose a risk to human health and therefore their dissemination in the food production chain needs to be assessed. METHODS: In this study, 450 boot swabs from chicken farms were analyzed for the presence of antimicrobial resistance with a focus on ß-lactams resistance in Acinetobacter species. RESULTS: Two ß-lactamase-encoding genes were first time identified in Acinetobacter lwoffii and Acinetobacter schindleri isolates. The deduced amino acid sequence of OXA-496 shared 93.8% identity with OXA-363. The second OXA-134-like enzyme, OXA-537, had the highest sequence identity (97.8%) with OXA-235 and OXA-237. CONCLUSIONS: The results of this study illustrate the occurrence of new OXA-134-like ß-lactamases, called OXA-496 and OXA-537, carrying strains of Acinetobacter lwoffii and Acinetobacter schindleri in chicken farm litter, and highlight the possible role of Acinetobacter as a reservoir of resistance genes.

Characteristics of Quinolone Resistance in Escherichia coli Isolates from Humans, Animals, and the Environment in the Czech Republic.

Escherichia coli is a common commensal bacterial species of humans and animals that may become a troublesome pathogen causing serious diseases. The aim of this study was to characterize the quinolone resistance phenotypes and genotypes in E. coli isolates of different origin from one area of the Czech Republic. E. coli isolates were obtained from hospitalized patients and outpatients, chicken farms, retailed turkeys, rooks wintering in the area, and wastewaters. Susceptibility of the isolates grown on the MacConkey agar with ciprofloxacin (0.05 mg/L) to 23 antimicrobial agents was determined. The presence of plasmid-mediated quinolone resistance (PMQR) and ESBL genes was tested by PCR and sequencing. Specific mutations in gyrA, gyrB, parC, and parE were also examined. Multilocus sequence typing and pulsed-field gel electrophoresis were performed to assess the clonal relationship. In total, 1050 E. coli isolates were obtained, including 303 isolates from humans, 156 from chickens, 105 from turkeys, 114 from the rooks, and 372 from wastewater samples. PMQR genes were detected in 262 (25%) isolates. The highest occurrence was observed in isolates from retailed turkey (49% of the isolates were positive) and inpatients (32%). The qnrS1 gene was the most common PMQR determinant identified in 146 (56%) followed by aac(6')-Ib-cr in 77 (29%), qnrB19 in 41 (16%), and qnrB1 in 9 (3%) isolates. All isolates with high level of ciprofloxacin resistance (>32 mg/L) carried double or triple mutations in gyrA combined with single or double mutations in parC. The most frequently identified substitutions were Ser(83)Leu; Asp(87)Asn in GyrA, together with Ser(80)Ile, or Glu(84)Val in ParC. Majority of these isolates showed resistance to beta-lactams and multiresistance phenotype was found in 95% isolates. Forty-eight different sequence types among 144 isolates analyzed were found, including five major clones ST131 (26), ST355 (19), ST48 (13), ST95 (10), and ST10 (5). No isolates sharing 100% relatedness and originating from different areas were identified. In conclusion, our study identified PMQR genes in E. coli isolates in all areas studied, including highly virulent multiresistant clones such as ST131 producing CTX-M-15 beta-lactamases.

Epidemiology of hospital-acquired pneumonia: Results of a Central European multicenter, prospective, observational study compared with data from the European region.

BACKGROUND: Hospital-acquired pneumonia (HAP) is associated with high mortality. In Central Europe, there is a dearth of information on the prevalence and treatment of HAP. This project was aimed at collecting multicenter epidemiological data on patients with HAP in the Czech Republic and comparing them with supraregional data. METHODS: This prospective, multicenter, observational study processed data from a database supported by a Czech Ministry of Health grant project. Included were all consecutive patients aged 18 and over who were admitted to participating intensive care units (ICUs) between 1 May 2013 and 31 December 2014 and met the inclusion criterion of having HAP. The primary endpoint was to analyze the relationships between 30-day mortality (during the stay in or after discharge from ICUs) and the microbiological etiological agent and adequacy of initial empirical antibiotic therapy in HAP patients. RESULTS: The group dataset contained data on 330 enrolled patients. The final validated dataset involved 214 patients, 168 males (78.5%) and 46 females (21.5%), from whom 278 valid lower airway samples were obtained. The mean patient age was 59.9 years. The mean APACHE II score at admission was 21. Community-acquired pneumonia was identified in 13 patients and HAP in 201 patients, of whom 26 (12.1%) had early-onset and 175 (81.8%) had late-onset HAP. Twenty-two bacterial species were identified as etiologic agents but only six of them exceeded a frequency of detection of 5% (Klebsiella pneumoniae 20.4%, Pseudomonas aeruginosa 20.0%, Escherichia coli 10.8%, Enterobacter spp. 8.1%, Staphylococcus aureus 6.2% and Burkholderia cepacia complex 5.8%). Patients infected with Staphylococcus aureus had significantly higher rates of early-onset HAP than those with other etiologic agents. The overall 30-day mortality rate for HAP was 29.9%, with 19.2% mortality for early-onset HAP and 31.4% mortality for late-onset HAP. Patients with late-onset HAP receiving adequate initial empirical antibiotic therapy had statistically significantly lower 30-day mortality than those receiving inadequate initial antibiotic therapy (23.8% vs 42.9%). Patients with ventilator-associated pneumonia (VAP) had significantly higher mortality than those who developed HAP with no association with mechanical ventilation (34.6% vs 12.7%). Patients having VAP treated with adequate initial antibiotic therapy had lower 30-day mortality than those receiving inadequate therapy (27.2% vs 44.8%). CONCLUSIONS: The present study was the first to collect multicenter data on the epidemiology of HAP in the Central European Region, with respect to the incidence of etiologic agents causing HAP. It was concerned with relationships between 30-day patient mortality and the type of HAP, etiologic agent and adequacy of initial empirical antibiotic therapy.

Incidence of fecal Enterobacteriaceae producing broad-spectrum beta-lactamases in patients with hematological malignancies.

AIM: Given the steadily increasing numbers of resistant bacteria, the frequency and severity of infections are on the rise. In patients with hematological malignancies, the treatment itself increases the risk of complicating bacterial infections. One important mechanisms of resistance is production of broad-spectrum beta-lactamases, increasingly detected not only in bacterial pathogens but also in bacteria contained in the normal microflora of the human body. The objectives of this study were determination and analysis of the prevalence of multiresistant ESBL- and AmpC-positive Enterobacteriaceae in the gastrointestinal tract (GIT) of patients with hematological malignancies. METHODS: For 3 months, rectal swabs were taken from patients with hematological malignancies and analyzed using chromogenic screening plates to isolate ESBL- and AmpC-producing Enterobacteriaceae. Beta-lactamase production was determined by phenotype tests and confirmed by detecting genes encoding ESBL and AmpC types. At the same time, ESBL- and AmpC-positive Enterobacteriaceae were isolated from clinical samples collected from patients with bacterial infection. RESULTS: Over the study period, fifteen patients (21%) of all patients treated at the Department of Hemato-Oncology were shown to have ESBL- or AmpC-positive Enterobacteriaceae in their GIT. Most frequently identified were ESBL-positive strains of Klebsiella pneumoniae and AmpC-positive strains of Citrobacter freundii. The ESBL enzymes were mainly of the CTX-M type. Isolates producing AmpC were found to contain genes for enzymes mainly from the CIT and DHA groups. CONCLUSION: The study identified patients diagnosed with urinary tract and bloodstream infections caused by ESBL-positive strain of Klebsiella pneumoniae and AmpC-positive strain of Enterobacter cloacae contained in the GIT microflora.

Carriage of ESBL- and AmpC-positive Enterobacteriaceae in the gastrointestinal tract of community subjects and hospitalized patients in the Czech Republic.

AIM: The study aimed at analyzing ESBL- and AmpC-positive Enterobacteriaceae in the gastrointestinal tracts of university hospital inpatients and persons from the Olomouc Region community, and comparing the results with data from 2007. METHODS: Bacteria were isolated from rectal swabs inoculated onto the ChromID(TM) ESBL selective medium (bioMérieux). Production of ESBL-type beta-lactamases was confirmed by the modified double-disk synergy test and AmpC enzyme production was detected by the AmpC disk test. ESBL- and AmpC-positive isolates were subjected to basic genetic analysis aimed at detecting the bla(TEM), bla(SHV), bla(CTX-M) and bla(AmpC) genes. RESULTS: Over the study period (1 March 2010 - 1 May 2010), a total of 1,279 rectal swabs (70.4% of community subjects) were analyzed on the above medium. The prevalence rates of ESBL-positive Enterobacteriaceae were 8.2% in hospitalized patients and 3.2% in community subjects. Production of the AmpC enzyme was detected in 1.1% of bacterial isolates from the community and in one (0.3%) hospital isolate. Among ESBL, the most frequent genes encoding enzymes were from the CTX-M-1-like genes. Detected AmpC beta-lactamases belonged to the CIT, DHA and EBC groups. CONCLUSION: When compared with the year 2007, the rates of carriers of ESBL-positive bacteria increased in both hospitalized patients (from 3% to 8%) and community subjects (from 1% to 3%) in 2010. Given the fact that production of extended-spectrum beta-lactamases is clinically significant, knowing the epidemiological situation is very important for selecting adequate antibiotic therapy.