Comparison of Virulence-Factor-Encoding Genes and Genotype Distribution amongst Clinical Pseudomonas aeruginosa Strains.

Pseudomonas aeruginosa is an opportunistic pathogen encoding several virulence factors in its genome, which is well-known for its ability to cause severe and life-threatening infections, particularly among cystic fibrosis patients. The organism is also a major cause of nosocomial infections, mainly affecting patients with immune deficiencies and burn wounds, ventilator-assisted patients, and patients affected by other malignancies. The extensively reported emergence of multidrug-resistant (MDR) P. aeruginosa strains poses additional challenges to the management of infections. The aim of this study was to compare the incidence rates of selected virulence-factor-encoding genes and the genotype distribution amongst clinical multidrug-sensitive (MDS) and MDR P. aeruginosa strains. The study involved 74 MDS and 57 MDR P. aeruginosa strains and the following virulence-factor-encoding genes: lasB, plC H, plC N, exoU, nan1, pilA, and pilB. The genotype distribution, with respect to the antimicrobial susceptibility profiles of the strains, was also analyzed. The lasB and plC N genes were present amongst several P. aeruginosa strains, including all the MDR P. aeruginosa, suggesting that their presence might be used as a marker for diagnostic purposes. A wide variety of genotype distributions were observed among the investigated isolates, with the MDS and MDR strains exhibiting, respectively, 18 and 9 distinct profiles. A higher prevalence of genes determining the virulence factors in the MDR strains was observed in this study, but more research is needed on the prevalence and expression levels of these genes in additional MDR strains.

Candida auris Infection in a Meningococcal Septicemia Survivor, Poland.

BACKGROUND: Candida auris is an emerging pathogen that constitutes a serious global health threat. It is difficult to identify without specific approaches, and it can be misidentified with standard laboratory methods, what may lead to inappropriate management. CASE PRESENTATION: We report, probably the first in Poland, C. auris isolation from blood cultures and wound swabs of a young male following meningococcal septicaemia, in February 2019. The patient had been previously hospitalized in the United Arab Emirates. The isolate was rapidly identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry and therefore clinicians were promptly informed on the alert pathogen isolation. The targeted antifungal treatment was successful and infection control measures seemed effective. ITS-based identification and subsequent whole genome sequencing showed that the C. auris isolate belongs to South Asian lineage (clade I). CONCLUSIONS: C. auris is able to cause outbreaks in healthcare settings. Therefore, it is important to quickly identify C. auris isolates in hospital settings so that healthcare facilities can take proper precautions to limit its spread.

Healthcare-associated infections in COVID-19 ICU patients - two-centre study.

OBJECTIVE: The aim of this retrospective study was to estimate the prevalence of healthcare-associated infections (HAI), microbiological data including resistance patterns and impact of HAI on patients' survival. METHODS: Two-centre study on 172 patients was performed. Medical records of patients hospitalized in the two COVID-19 intensive care units (ICU) localized in Bydgoszcz between 1 October 2020 and 30 March 2021 were analysed retrospectively. Data collection included demographics, microbiological, clinical variables, and patient outcome. All infections were defined according to the HAI-Net ICU protocol of the European Centre for Disease Prevention and Control (ECDC). Detailed data concerning bloodstream infection (BSI), pneumonia (PN) and urinary tract infection (UTI) were collected. RESULTS: In 97 patients (56.4%), 138 HAI cases were identified. Patients with HAI statistically more often had been administered antimicrobial therapy prior to the admission to ICU (59.8% vs. 34.7%, p < 0.05), and needed catecholamines during hospitalization (93.8% vs. 70.7%, p < 0.001). The risk of HAI increased by 50% if antimicrobial therapy had been applied before the admission to ICU, and was three times higher if during the hospitalization in ICU catecholamines infusion was needed. Mortality was higher in patients diagnosed with HAI (72.2% vs. 65.3%) but the difference was not statistically significant (p = 0.34). CONCLUSIONS: Further investigation of co-infections in critically ill patients with COVID-19 is required in order to identify HAI risk factors, define the role of empiric antimicrobial therapy and proper prevention strategies.

Bacteraemia Caused by Probiotic Strains of Lacticaseibacillus rhamnosus-Case Studies Highlighting the Need for Careful Thought before Using Microbes for Health Benefits.

Lactic acid bacteria belonging to Lactobacillus spp. and Lacticaseibacillus spp. are a natural part of fermented milk and other food products, probiotic supplements and human microbiota. They mainly belong to mucosal microflora, especially oral, vaginal and intestinal. Lacticaseibacillus spp. strains included in probiotics are generally characterised as safe microorganisms, and the species are concerned bacteria with very low pathogenic potential. However, infections caused by Lactobacillus spp. and Lacticaseibacillus spp., including bacteraemia and endocarditis, occur occasionally. The aim of the study was to present two cases of bacteraemia due to Lacticaseibacillus rhamnosus associated with the use of a probiotic product. It afflicted patients in intensive care units. The investigation was preliminarily based on clinical and microbiological recognition of the cases. The initial observation was laboratory confirmed with the application of pulsed-field gel electrophoresis (PFGE) results. Identical PFGE patterns were obtained for the evaluated strains and the strains derived from a commercially available probiotic that was administered to those patients. The increasing number of studies describing opportunistic infections due to probiotic strains of Lacticaseibacillus spp. should result in verifying the safety of probiotic formulations used in immunocompromised patients and forming detailed guidelines for the use of probiotics among patients from several risk groups.

Mortality Following Clostridioides difficile Infection in Europe: A Retrospective Multicenter Case-Control Study.

We aimed to describe the clinical presentation, treatment, outcome and report on factors associated with mortality over a 90-day period in Clostridioides difficile infection (CDI). Descriptive, univariate, and multivariate regression analyses were performed on data collected in a retrospective case-control study conducted in nine hospitals from seven European countries. A total of 624 patients were included, of which 415 were deceased (cases) and 209 were still alive 90 days after a CDI diagnosis (controls). The most common antibiotics used previously in both groups were ß-lactams; previous exposure to fluoroquinolones was significantly (p = 0.0004) greater in deceased patients. Multivariate logistic regression showed that the factors independently related with death during CDI were older age, inadequate CDI therapy, cachexia, malignancy, Charlson Index, long-term care, elevated white blood cell count (WBC), C-reactive protein (CRP), bacteraemia, complications, and cognitive impairment. In addition, older age, higher levels of WBC, neutrophil, CRP or creatinine, the presence of malignancy, cognitive impairment, and complications were strongly correlated with shortening the time from CDI diagnosis to death. CDI prevention should be primarily focused on hospitalised elderly people receiving antibiotics. WBC, neutrophil count, CRP, creatinine, albumin and lactate levels should be tested in every hospitalised patient treated for CDI to assess the risk of a fatal outcome.

Education and training programmes for infection prevention and control professionals: mapping the current opportunities and local needs in European countries.

BACKGROUND: Studies have repeatedly highlighted the need for homogenisation of training content and opportunities in infection prevention and control (IPC) across European countries. OBJECTIVES: To map current training opportunities for IPC professionals, define local needs and highlight differences, across 11 European countries (Cyprus, France, England, Germany, Greece, Italy, Netherlands, Poland, Romania, Spain, Switzerland). SOURCES: From July 2018 to February 2019, IPC experts directly involved in IPC training and education in their countries and/or internationally were invited to complete a prespecified set of questions in order to provide a detailed description of IPC training opportunities and needs in their country. CONCLUSIONS: IPC training among nurses and doctors varies greatly across countries, with differences in content and type of training (e.g., standardised curriculum, educational programme, clinical experience) duration, as well as in assessment and recognition/accreditation. The observed heterogeneity in IPC training between European countries can be eliminated through establishment of interdisciplinary region-wide training programmes, with common learning objectives, shared know-how and supported by national and international professional bodies.

Fulminant mucormycosis after a traffic accident: a case report.

Mucormycosis is a rare fungal infection in immunocompetent patients, whereas in immunocompromised, it may be systemic and disseminated infection associated with high mortality. Mucormycosis is one of the most rapidly progressing and fulminant forms of fungal infections; Mucor circinelloides is rarely isolated species, also from immunocompromised patients. The reported case of mucormycosis after a traffic accident indicates that it may be the result of a contamination of wound by M. circinelloides coming from the environment. The fungal strain was identified by phenotypic methods and confirmed by molecular methods. Etest method was used for susceptibility testing of the fungal strain. No mycotoxins were detected in the analyzed sample. The infection was successfully treated with amphotericin B, but amputation of the lower limb was necessary.

Prevalence of healthcare-associated infections, estimated incidence and composite antimicrobial resistance index in acute care hospitals and long-term care facilities: results from two European point prevalence surveys, 2016 to 2017.

Point prevalence surveys of healthcare-associated infections (HAI) and antimicrobial use in the European Union and European Economic Area (EU/EEA) from 2016 to 2017 included 310,755 patients from 1,209 acute care hospitals (ACH) in 28 countries and 117,138 residents from 2,221 long-term care facilities (LTCF) in 23 countries. After national validation, we estimated that 6.5% (cumulative 95% confidence interval (cCI): 5.4-7.8%) patients in ACH and 3.9% (95% cCI: 2.4-6.0%) residents in LTCF had at least one HAI (country-weighted prevalence). On any given day, 98,166 patients (95% cCI: 81,022-117,484) in ACH and 129,940 (95% cCI: 79,570-197,625) residents in LTCF had an HAI. HAI episodes per year were estimated at 8.9 million (95% cCI: 4.6-15.6 million), including 4.5 million (95% cCI: 2.6-7.6 million) in ACH and 4.4 million (95% cCI: 2.0-8.0 million) in LTCF; 3.8 million (95% cCI: 3.1-4.5 million) patients acquired an HAI each year in ACH. Antimicrobial resistance (AMR) to selected AMR markers was 31.6% in ACH and 28.0% in LTCF. Our study confirmed a high annual number of HAI in healthcare facilities in the EU/EEA and indicated that AMR in HAI in LTCF may have reached the same level as in ACH.

Surveillance of Antibiotic Prescribing in Intensive Care Units in Poland.

Antibiotic use and microbial resistance in health care-associated infections are increasing globally and causing health care problems. Intensive Care Units (ICUs) represent the heaviest antibiotic burden within hospitals, and sepsis is the second noncardiac cause of mortality in ICUs. Optimizing appropriate antibiotic treatment in the management of the critically ill in ICUs became a major challenge for intensivists. We performed a surveillance study on the antibiotic consumption in 108 Polish ICUs. We determined which classes of antibiotics were most commonly consumed and whether they affected the length of ICU stay and the size and category of the hospital. A total of 292.389 defined daily doses (DDD) and 192.167 patient-days (pd) were identified. Antibiotic consumption ranged from 620 to 3960 DDD/1000 pd. The main antibiotic classes accounted for 59.6% of the total antibiotic consumption and included carbapenems (17.8%), quinolones (14%), cephalosporins (13.7%), penicillins (11.9%), and macrolides (2.2%), respectively, whereas the other antibiotic classes accounted for the remainder (40.4%) and included antifungals (34%), imidazoles (20%), aminoglycosides (18%), glycopeptides (15%), and polymyxins (6%). The most consumed antibiotic classes in Polish ICUs were carbapenems, quinolones, and cephalosporins, respectively. There was no correlation between antibiotic consumption in DDD/1000 patient-days, mean length of ICU stay, size of the hospital, size of the ICU, or the total amount of patient-days. It is crucial that surveillance systems are in place to guide empiric antibiotic treatment and to estimate the burden of resistance. Appropriate use of antibiotics in the ICU should be an important public health care issue.

Use of ECDC PPS HAI&AU data to evaluate adherence to national guidelines for antimicrobial treatment of community-­acquired pneumonia

INTRODUCTION    Point prevalence surveys are widely described as a useful tool for evaluating antimicrobial policy and adherence to guidelines. OBJECTIVES    We aimed to investigate if data from the European Centre for Disease Prevention and Control (ECDC) point prevalence survey of healthcare­associated infections and antimicrobial use (PPS HAI&AU) canbe used to evaluate adherence to national guidelines for the treatment of community­acquired pneumonia (CAP) and to analyze the quality of treatment regimens. PATIENTS AND METHODS    Data for 72 698 patients were collected in Poland between the years 2012 and 2015 according to the ECDC Protocol v.4.2. CAP was an indication for antimicrobial treatment in 3608patients. Patients hospitalized longer than 48 hours were excluded. A total of 667 patients met the inclusion criteria, and 79 regimens were recorded and evaluated as concordant or discordant with the guidelines. Afterwards, 7 experts scored the regimens from 1 to 5. The averages were calculated, and the results below 3.0 were considered as not optimal and those of 3.0 or higher-as optimal. Coherence of the experts' scores was evaluated. RESULTS    Of all patients, 153 (22.8%) were treated exactly according to the guidelines. Nineteen regimens (24.0%) were optimal but discordant with the guidelines; they were administered to 346 patients (51.9%). The remaining 50 regimens (63.3%) were evaluated as discordant and not optimal and were used in 169 patients(25.3%). The correlation results of the experts' scores were significant. CONCLUSIONS    ECDC PPS HAI&AU data can be efficiently used to assess adherence to guidelines. Despite low adherence (22.8%), almost 75% of patients received optimal antimicrobial treatments. Actions promoting the guidelines and time­series studies analyzing improvement of adherence should be considered.

Healthcare associated bloodstream infections in Polish hospitals: prevalence, epidemiology and microbiology-summary data from the ECDC Point Prevalence Survey of Healthcare Associated Infections 2012-2015.

Aggregated data from the Polish Point Prevalence Survey of Healthcare Associated Infections and Antimicrobial Use (PPS HAI&AU) collected between 2012 and 2015 were used to describe the epidemiology of healthcare associated bloodstream infections (BSI) in Polish hospitals, in order to assess the rationale for introducing a BSI surveillance programme in our country and analyse selected risk factors. Data were collected according to the ECDC PPS HAI&AU protocol. Within four years, records for 71,039 patients were collected in 36 (2012), 32 (2013), 112 (2014), and 158 (2015) hospitals; representativeness was evaluated as good in 2012-2013, and excellent from 2014. HAI was found in 4,258 of these patients; laboratory confirmed BSI, including catheter related infections (CRI), and neonatal BSI accounted for 7.7% (329 cases). A representative control group was selected during a random selection process. Out of 329 cases of BSI, 48.9% were associated with vascular access, and 70.8% of them met the criteria of (CRI). The most frequently isolated microorganisms were Staphylococci with 150 isolates (45.6%). Most of them were coagulase-negative (64.4%) that usually caused CRI. Out of 53 S. aureus isolates 24.5% were methicillin-resistant. Enterobacteriaceae were responsible for 31.3% of BSI (n = 103), 50.0% of them were resistant to third generation cephalosporins and 6 (5.8%) to carbapenems. Since little is known about the epidemiology of BSI in Poland, introduction of a countrywide surveillance programme based on incidence is justified, in order to create national prevention initiatives based on local epidemiology, as well as bundle focusing on prevention of CRI.

The Prevalence of Exoenzyme S Gene in Multidrug-Sensitive and Multidrug-Resistant Pseudomonas aeruginosa Clinical Strains.

Pseudomonas aeruginosa rods are one of the most commonly isolated microorganisms from clinical specimens, usually responsible for nosocomial infections. Antibiotic-resistant P. aeruginosa strains may present reduced expression of virulence factors. This fact may be caused by appropriate genome management to adapt to changing conditions of the hospital environment. Virulence factors genes may be replaced by those crucial to survive, like antimicrobial resistance genes. The aim of this study was to evaluate, using PCR, the occurrence of exoenzyme S-coding gene (exoS) in two distinct groups of P. aeruginosa strains: 83 multidrug-sensitive (MDS) and 65 multidrug-resistant (MDR) isolates. ExoS gene was noted in 72 (48.7%) of the examined strains: 44 (53.0%) MDS and 28 (43.1%) MDR. The observed differences were not statistically significant (p = 0.1505). P. aeruginosa strains virulence is rather determined by the expression regulation of the possessed genes than the difference in genes frequency amongst strains with different antimicrobial susceptibility patterns.

Transdifferentiation of Bone Marrow Mesenchymal Stem Cells into the Islet-Like Cells: the Role of Extracellular Matrix Proteins.

Pancreatic islet implantation has been recently shown to be an efficient method of treatment for type 1 diabetes. However, limited availability of donor islets reduces its use. Bone morrow would provide potentially unlimited source of stem cells for generation of insulin-producing cells. This study was performed to evaluate the influence of extracellular matrix proteins like collagen, laminin, and vitronectin on bone marrow mesenchymal stem cells (BM-MSCs) transdifferentiation into islet-like cells (ILCs) in vitro. To our knowledge, this is the first report evaluating the importance of vitronectin in transdifferentiation of BM-MSCs into ILCs. Rat BM-MSCs were induced to ILCs using four-step protocol on plates coated with collagen type IV, laminin type I and vitronectin type I. Quantitative real-time PCR was performed to detect gene expression related to pancreatic ß cell development. The induced cells expressed islet-related genes including: neurogenin 3, neurogenic differentiation 1, paired box 4, NK homeobox factor 6.1, glucagon, insulin 1 and insulin 2. Laminin but not collagen type IV or vitronectin enhanced expression of insulin and promoted formation of islet-like structures in monolayer culture. Laminin triggered transdifferentiation of BM-MSCs into ILCs.

Impact of fructose diet and renal failure on the function of pancreatic islets.

OBJECTIVES: This study was designed to evaluate the impact of fructose-rich diet and chronic kidney disease (CKD) on the in vitro function of pancreatic islets. METHODS: Fifty-four rats were divided into 3 equal groups as follows: control, rats with CKD 1/2 that underwent surgical uninephrectomy, and rats with CKD 5/6 that underwent uninephrectomy and kidney cortex mass resection. Each group was further assigned to 3 diet protocols--regular diet, regular diet with 10% fructose (F10), and 60% fructose-rich diet (F60). After 8 weeks of insulin administration, C-peptide, glycated hemoglobin level, serum urea nitrogen, creatinine clearance, and homeostasis model assessment of insulin resistance were evaluated. Static glucose insulin stimulation test of isolated pancreatic islets and histologic analysis of pancreatic tissue were performed. RESULTS: The F10 diet increased the levels of insulin and C-peptide in all groups. Homeostasis model assessment of insulin resistance was increased in all animals fed with fructose. The elevated levels of creatinine and diminished creatinine clearance were detected in CKD 5/6 rats fed with 60% fructose-rich diet. The F10 diet resulted in high levels of serum insulin and C-peptide and glucose-stimulated insulin secretion. Fructose-rich diet increased the islet size and number, with irregular morphology and exocrine tissue fibrosis. CONCLUSIONS: The fructose-rich diet accelerates the progression of CKD and affects the pancreatic islet function.

Does Toxoplasma gondii infection affect cognitive function? A case control study.

According to studies, latent Toxoplasma gondii infection may affect several functions of the human brain. Here we search for the association between latent toxoplasmosis and cognitive performance. We tested 70 individuals for latent T. gondii infection. There were 26 Toxoplasma-infected subjects and 44 Toxoplasma-free subjects. Within these two groups we assessed cognitive performance using a set of standardized, widely recognized neuropsychological tests: Trail Making Test, Stroop Test, Verbal Fluency Test, Digit Span Test and N-back test. The relationship between chronic toxoplasmosis and cognitive performance was assessed, with adjustment for age and sex. Patients with latent toxoplasmosis performed worse on one neuropsychological test, N-back Test--percentage of correct answers (beta -8.08; 95% CI - 15.64 to -0.53; p < 0.05) compared to seronegative patients. However, after adjustment for age and sex, no statistically significant associations between latent toxoplasmosis and the scores on any cognitive tests were noticed. As statistically significant relationship was not observed, this study does not confirm that chronic latent T. gondii infection affects cognition.

Evaluation of different methods for detection of metallo-beta-lactamases in Pseudomonas aeruginosa clinical isolates.

Metallo-beta-lactamases (MBLs) produced by Pseudomnonas aeruginosa are a serious threat due to their ability to be transmitted between the same as well as different bacterial species. Different methods are applied in the clinical laboratory to detect MBLs. The aim of this study was to compare 4 phenotypic methods and a PCR assay for their ability to detect MBLs in clinical isolates of carbapenem-resistant P. aeruginosa strains. The study embraced a total of 70 carbapenem-resistant P. aeruginosa strains isolated in The Department of Microbiology of Dr. A. Jurasz University Hospital in Bydgoszcz. The highest percentage (42.9%) of the strains were isolated from Intensive Care Unit patients, mainly from urine samples (31.4%). Methods used in this study were: double-disc synergy tests in two combinations: using ceftazidime with 2-mercaptopropionic acid and imipenem with EDTA, differences in inhibition zone diameters between discs with imipenem/EDTA and imipenem, Etest MBL (AB Biodisk) and molecular amplification of bla(IMP) and bla(VIM) genes responsible for producing MBLs, using PCR assay. The lowest percentage (1.4%) of positive results in detection of MBLs was obtained using PCR assay, the highest (72.9%) by double-disc synergy tests with imipenem and EDTA, but the specificity of this method may be low.

Antimicrobial susceptibility of metallo-beta-lactamase positive and negative Klebsiella pneumoniae strains isolated from intensive care unit patients.

Carbapenem resistance in Gram-negative bacteria is a worldwide increasing and one of the most disturbing problems, given these antibiotics are drugs of choice in the treatment of infections caused by extended-spectrum-beta-lactamase producing strains. In this study the antibiotic susceptibility of metallo-beta-lactamase-positive and negative Klebsiella pneumoniae strains isolated from intensive care unit (ICU) patients was evaluated. The presence of genes encoding MBLs was determined with a commercial kit hyplex MBL ID (BAG HEALTH CARE). The MBL-producing isolates were the first K. pneumoniae isolates of this kind identified in Poland. It seems that methods for detecting MBLs in Enterobacteriaceae should be included in contemporary standards of microbiological diagnostics in the country.

Reduced expression of virulence factors in multidrug-resistant Pseudomonas aeruginosa strains.

MDR Pseudomonas aeruginosa strains are isolated from clinical specimens with increasing frequency. It seems that acquiring genes which determine antibiotic resistance usually comes at a biological cost of impaired bacterial physiology. There is no information on investigations comparing phenotypic differences in MDR and MDS P. aeruginosa strains in literature. The study included 150 clinical P. aeruginosa isolates (75 classified as MDS and 75 as MDR). PFGE analysis revealed five pairs of identical isolates in the group of MDR strains and the results obtained for these strains were not included in the statistical analyses. MDR strains adhered to polystyrene to a lesser extent than MDS strains. The growth rate in the liquid medium was significantly lower for MDR strains. Detectable amounts of alginate were present in the culture supernatants of seven MDS and six MDR strains. The MDR P. aeruginosa strains which were investigated produced significantly lower amounts of extracellular material binding Congo Red, lower lipolytic, elastase, LasA protease, phospholipase C activity and pyocyanin quantity in culture supernatants when compared with MDS strains. No significant differences were observed between MDR and MDS strains in proteolytic activity. In conclusion, the MDR P. aeruginosa strains have impaired virulence when compared to MDS strains.

[Relatedness of methicillin-resistant coagulase-negative staphylococci]. / Pokrewienstwo meticylinoopornych szczepów gronkowców koagulazo-ujemnych.

Many identification and relatedness studies methods had been commonly used for epidemiological studies in microbiological laboratories. Apart from phenotypic methods, genotypic are also often used. The aim of this study was to compare, obtained by PFGE chromosomal DNA patterns of methicillin-resistant S. epidermidis strains isolated from clinical material. 46 methicillin-resistant S. epidermidis strains were included in this study. Most of them were isolated from wound swabs (65.2%) and catheters (19.6%) from different surgical clinics (76.1%). To identify strains and receive biochemical profiles, ID 32 Staph tests and GPI cards of Vitek 1 system were used. Pulsed-field gel electrophoresis and Tenover et al. interpretation were used to compare chromosomal DNA patterns of examined strains. 44 and 42 PFGE patterns of chromosomal DNA were received, using visual interpretation classifying two pairs of strains as the same, two pairs as closely related and three pairs as probably related. Strains classified as identical and similar in visual evaluation were indistinguishable in Molecular Analyst DST interpretation, probably due to tolerance in bands location pattern. Strains probably related in visual interpretation represent at least 96% similarity in Molecular Analyst DST but different susceptibility and biochemical profiles obtained by ID 32 Staph and Vitek 1. PFGE analysis had foremost capacity to distinguish methicillin-resistant S. epidermidis strains using visual interpretation and Molecular Analyst DST (Bio-Rad) program and seems to be useful method in epidemiological studies. Strains with the same PFGE pattern, had different susceptibility and biochemical profiles.