Antibacterial Therapy by Ag+ Ions Complexed with Titan Yellow/Congo Red and Albumin during Anticancer Therapy of Urinary Bladder Cancer.

According to the World Health Organization report, the increasing antibiotic resistance of microorganisms is one of the biggest global health problems. The percentage of bacterial strains showing multidrug resistance (MDR) to commonly used antibiotics is growing rapidly. Therefore, the search for alternative solutions to antibiotic therapy has become critical to combat this phenomenon. It is especially important as frequent and recurring infections can cause cancer. One example of this phenomenon is urinary tract infections that can contribute to the development of human urinary bladder carcinoma. This tumor is one of the most common malignant neoplasms in humans. It occurs almost three times more often in men than in women, and in terms of the number of cases, it is the fifth malignant neoplasm after prostate, lung, colon, and stomach cancer. The risk of developing the disease increases with age. Despite the improvement of its treatment methods, the current outcome in the advanced stages of this tumor is not satisfactory. Hence, there is an urgent need to introduce innovative solutions that will prove effective even in the advanced stage of the disease. In our study, a nanosystem based on ionic silver (Ag+) bound to a carrier-Titan yellow (TY) was analyzed. The possibility of binding the thus formed TY-Ag system to Congo red (CR) and albumin (BSA) was determined. TY-Ag binding to CR provides for better nanosystem solubility and enables its targeted intracellular transport and binding to immune complexes. The binding of TY-Ag or CR-TY-Ag to albumin also protects the system against the uncontrolled release of silver ions. It will also allow the delivery of silver in a targeted manner directly to the desired site in the case of intravenous administration of such a system. In this study, the MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) values of the TY-Ag or BSA-TY-Ag systems were determined in two reference strains (Escherichia coli and Staphylococcus aureus). The paper presents nanosystems with a size of about 40-50 nm, with an intense antibacterial effect obtained at concentrations of 0.019 mM. We have also discovered that TY-Ag free or complexed with BSA (with a minimal Ag+ dose of 15-20 µM) inhibited cancer cells proliferation. TY-Ag complex diminished migration and effectively inhibited the T24 cell viability and induced apoptosis. On the basis of the obtained results, it has been shown that the presented systems may have anti-inflammatory and antitumor properties at the same time. TY-Ag or BSA-TY-Ag are new potential drugs and may become in future important therapeutic compounds in human urinary bladder carcinoma treatment and/or potent antimicrobial factors as an alternative to antibiotics.

[Molecular characterization of Staphylococcus aureus strains isolated from surgical site infections of patients in southern Poland].

INTRODUCTION: The aim of the study was a molecular characterization of Staphylococcus aureus strains isolated from surgical site infections (SSIs) from patients in southern Poland, undergoing different surgical procedures, together with evaluation of the prevalence of antimicrobial resistance and the presence of virulence factors. MATERIALS AND METHODS: In this laboratory-based, multicenter study, non-repetitive 162 samples from SSI were collected from hospitalized patients (12 hospitals, n=139) or outpatients (n=23) in southern Poland between January 1 and December 31, 2013. In all S. aureus isolates, we investigated antimicrobial susceptibility, the presence of selected virulence genes (lukE, pvl, tsst-l and eta), and also conducted spa typing. RESULTS: Patients with SSI had a median age of 61 years; 54.9% were male. Prevalence of MRSA (29 strains, 17.9%) SSI per surgery type was 8.7% in orthopaedic, 17.7% in general and 42.9% in vascular surgery. Over 20% of strains were resistant for erythromycin (27.2%), clindamycin (23.5%). No resistance was found for linezolid, glycopeptides or tigecycline. Gene of Leukocidin (lukE) was the most frequently found gene. Spa typing identified 10 spa types; the two dominant types were t003 (41.4%) and t138(17.2%). CONCLUSIONS: The results show that after vascular surgery, there was an unexpectedly high prevalence of MRSA in SSIs in southern Poland. Conversely, the prevalence of MRSA was unexpectedly low following orthopaedics procedures. The surprisingly observation was the low virulence of the S. aureus strains among older patients.

Bacteria contamination of touch surfaces in Polish hospital wards.

BACKGROUND: The objective of the study has been to evaluate the pathogenic bacteria contamination of touch surfaces in hospital wards. MATERIAL AND METHODS: Samples were taken from frequently touched surfaces in the hospital environment in 13 units of various types. Culturing was carried out on solid blood agar and in growth broth (tryptic soy broth - TSB). Species identification was performed using the analytical profile index (API) biochemical testing and confirmed with matrix assisted laser desorption ionization time-flight mass spectrometry (MALDI-TOF-MS) system. RESULTS: The total of 161 samples were taken for the study. Fifty-two of them, after 24 h of culture on a solid medium, demonstrated bacterial growth and further 60 samples had growth after prior multiplication in TSB. Overall, 69.6% of samples exhibited growth of 19 bacterial species. Pathogenic species - representing indicator organisms of efficiency of hospital cleaning - was demonstrated by 21.4% of samples. Among them Acinetobacter spp., Enterocococci spp. and Staphylococcus aureus were identified. Coagulase-negative staphylococci (CNS) were predominant. The proportion of various groups of bacteria significantly varied in respective hospitals, and in various types of wards. Disturbing observation is a large proportion of resistance of isolated CNS strains as a potential reservoir of resistance genes. CONCLUSIONS: The results show that touch surfaces in hospital units are contaminated by both potentially pathogenic and pathogenic bacterial species. In connection with the reported, also in Poland, frequent omission or incorrect execution of hand hygiene by hospital staff, and probably patients, touch surfaces still constitute important reservoir of pathogenic bacteria. Improving hand hygiene compliance of health-care workers with recommendations is necessary for increasing biological safety of hospital environment. Med Pr 2017;68(3):459-467.

Epidemiology, antibiotic consumption and molecular characterisation of Staphylococcus aureus infections--data from the Polish Neonatology Surveillance Network, 2009-2012.

BACKGROUND: Our aim was to determine and characterize S. aureus (SA) isolated from infections in newborns for antibiotic resistance, virulence factors, genotypes, epidemiology and antibiotic consumption. METHODS: Prospective surveillance of infections was conducted. Data about antibiotic treatment were analyzed. Antimicrobial susceptibility was assessed. PCR amplification was used to detect resistance and virulence genes. Typing methods such as PFGE, spa-typing and SCCmec were used. RESULTS: SA was found to be associated with 6.5% of infections. Methicillin-Resistant Staphylococcus aureus accounted for 32.8% of SA-infections. An incidence of MRSA-infections was 1.1/1000 newborns. MRSA-infections were diagnosed significantly earlier than MSSA-infections in these newborns (14th day vs. 23rd day (p=0.0194)). MRSA-infections increased the risk of newborn's death. Antibiotic consumption in both group was similar, but a high level of glycopeptides-usage for MSSA infections was observed. In the MRSA group, more strains were resistant to erythromycin, clindamycin, gentamicin and amikacin than in the MSSA group. Hla gene was present in 93.9% of strains, and seg and sei in 65.3% of strains, respectively. One dominant clone was found among the 14 MRSA isolates. Fifteen strains belonging to SCCmec type IV were spa-t015 and one strain belonging to SCCmec type V was spa-t011. CONCLUSIONS: Results obtained in the study point at specific epidemiological situation in Polish NICU (more detailed studies are recommended). High usage of glycopeptides in the MSSA infections treatment indicates the necessity of antimicrobial stewardship improvement and introducing molecular screening for early identification of infections.

Bloodstream Infections due to Enterobacteriaceae Among Neonates in Poland--Molecular Analysis of the Isolates.

Bloodstream infections (BSIs) are associated with a significantly increased risk of fatality. No report has been found about the molecular epidemiology of Enterobacteriaceae causing BSI in neonates in Poland. The aim of this work was to determine the antibiotic resistance profiles, virulence gene prevalence, the epidemiological and genetic relationships among the isolates from Enterobacteriaceae causing BSI in neonates with birth weight < 1501 g. Antimicrobial susceptibility testing was performed. PCR was performed to identify the presence of common beta-lactamase genes, virulence genes. PFGE and MLST were performed. The surveillance group contained 1,695 newborns. The incidence rate for BSIs was 5.9%, the fatality rate 15%. The most common species were Escherichia coli (n = 24) and Klebsiella pneumoniae (n = 16). CTX-M-15 was found in 6 E. coli, 8 K. pneumoniae, 1 Enterobacter cloacae strains. Among E. coli fimH (83.3%), ibeA (37.5%), neuC (20.8%) were the most frequent. PFGE demonstrated unique pulsotypes among E. coli. E. coli ST131 clone was found in 7 E. coli strains. PFGE of 16 K. pneumoniae strains showed 8 pulsotypes. Five isolates from one NICU belonged to one clone. MLST typing revealed 7 different ST with ST336 as the most prevalent. This study provides information about resistance, virulence and typing of Enterobacteriaceae strains causing BSI among neonates. E. coli and Klebsiella spp. isolated in this study have completely different epidemiology from each other.

The general status of patients and limited physical activity as risk factors of Methicillin-resistant Staphylococcus aureus occurrence in long-term care facilities residents in Krakow, Poland.

BACKGROUND: The aim of this study was to investigate the epidemiology and resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from long-term care facilities (LTCF) residents and to analyze the potential risk factors for MRSA occurrence, defined as MRSA colonization and/or infection. METHODS: Point prevalence (PPS) and prospective incidence continuous study (CS) was carried out on a group of 193 residents in 2009-2010. RESULTS: Overall MRSA occurred (with or without infection) among 17.6% of residents. There was 16 cases of infections with SA aetiology, of which 10 (58.8%) were caused by MRSA. The MRSA prevalence in PPS was 12.9%, in CS infection incidence rate was 5.2%. Factors associated with MRSA occurrence were: general status of patients, limited physical activity, wound infections (odds ratio, OR 4.6), ulcers in PPS (OR 2.1), diabetes (OR 1.6), urinary catheterization (OR 1.6) and stool incontinence (OR 1.2). CONCLUSIONS: Our data indicate a need for screening of MRSA before hospitalization or transfer to rehabilitation centres, especially in a group of residents with limitations in physical activity - i.e. with the highest risk of MRSA. Results also suggest the need for contact precautions in patients with high risk of MRSA occurrence, only. Focus on the high-risk population might be a solution for the cost-effective surveillance.

Virulence and Antibiotic Resistance of Pseudomonas aeruginosa Isolated from Patients with Urinary Tract Infections in Southern Poland.

BACKGROUND: The aim of this study was to analyze the resistance and virulence of Pseudomonas aeruginosa strains causing urinary tract infections in in- and outpatients in Southern Poland. METHODS: The study included 83 inpatients and 66 outpatients; 36.9% were female. RESULTS: Monomicrobial infections accounted for 74.5%; polymicrobial infections occurred more frequently among inpatients (odds ratio, OR = 4.32, p = 0.0008). exoS and lasB were detected in 90 and 74% of isolates, respectively. aprA was present in 66%, pilB in 5% and pilA in 23% of isolates. Isolates from adults were more frequently resistant to fluoroquinolones (OR = 0.37, p = 0.029). Twenty-nine isolates were classified as multidrug resistant and 12 as extremely drug resistant, which occurred less frequently in patients <17 years (OR = 0.18, p = 0.024). Nine metallo-ß-lactamase-positive isolates were identified. blaSHV was present in 10, blaTEM in 6, blaOXA-10 in 3 and blaVIM-2 in 3 isolates. CONCLUSION: Antibiotic selection should be based on the knowledge of local antimicrobial susceptibilities to maximize the benefit for patients and minimize the risk of drug resistance.

Mode of delivery and other risk factors for Escherichia coli infections in very low birth weight infants.

BACKGROUND: Infections in newborns remain one of the most significant problems in modern medicine. Escherichia coli is an important cause of neonatal bloodstream and respiratory tract infections and is associated with high mortality. The aim of our study was to investigate the epidemiology of E. coli infection in Polish neonatal intensive care units (NICUs) and resistance to antibiotics, with particular reference to the safety of very low birth weight infants. METHODS: Continuous prospective infection surveillance was conducted in 2009-2012 in five NICUs, including 1,768 newborns whose birth weight was <1.5 kg. Escherichia coli isolates from different diagnostic specimens including blood, tracheal/bronchial secretions and others were collected. All isolates were tested using disk diffusion antimicrobial susceptibility methods. Pulsed-field gel electrophoresis was used to determine the possible horizontal transfer of E. coli among patients. RESULTS: The incidence of E. coli infections was 5.4% and 2.0/1,000 patient-days. The occurrence of E. coli infections depended significantly on the NICU and varied between 3.9% and 17.9%. Multivariate analysis that took into account the combined effect of demographic data (gender, gestational age and birth weight) and place of birth showed that only the place of hospitalisation had a significant effect on the E. coli infection risk. The highest levels of resistance among all E. coli isolates were observed against ampicillin (88.8%) and amoxicillin/clavulanic acid (62.2%). Among E. coli isolates, 17.7% were classified as multidrug resistant. Escherichia coli isolates showed different pulsotypes and dominant epidemic clones were not detected. CONCLUSIONS: Our data indicate that antibiotic prophylaxis in the presence of symptoms such as chorioamnionitis and premature rupture of membranes did not help reduce the risk of E. coli infection. Multivariate analysis demonstrated only one significant risk factor for E. coli infection among infants with a birth weight <1.5 kg, that is, the impact of the NICU, it means that both neonatal care and care during pregnancy and labour were found to be significant.

Molecular characterization and drug resistance of Escherichia coli strains isolated from urine from long-term care facility residents in Cracow, Poland.

BACKGROUND: The aim of this study was to assess the prevalence of multidrug-resistant Escherichia coli and extended-spectrum â-lactamases (ESBL) pathogens isolated from asymptomatic bacteriuria and urinary tract infections (UTIs), and the relationship between the phylogeny, antimicrobial resistance, and virulence among isolates in residents of 3 long-term care facilities (LTCF) in Krakow, Poland. MATERIAL AND METHODS: This was point prevalence study and prospective infection control in a group of 217 people. Urine samples were examined with standard microbiological methods and screened for the presence of blaCTX-M, blaSHV, and blaTEM. E. coli isolates were screened for 6 common virulence factors (VFs) and classified according to the rapid phylogenetic grouping technique. RESULTS: Among all the strains tested, 14 isolates (13.9%) expressed ESBL activity. A significant proportion of isolates were resistant to ciprofloxacin (32.7%, n=33). Resistance to trimethoprim/sulfamethoxazole was identified among 45 isolates (44.5%). Independent risk factors for the presence of an ESBL-producing strain were: UTI, urinary and/or fecal incontinence, bedridden, and low values of the Barthel and Katz Indexes. Gene sequencing identified 8 blaCTX-M-15, 1 blaCTX-M-3, 9 blaTEM-1, and 1 blaSHV-12. Among E. coli, no relationship between number of VF genes and phylogeny was found. The most prevalent virulence factor was fimH (82.1%). CONCLUSIONS: The findings of this study emphasize the need for further research on the epidemiology of multi-drug resistant organisms (MDRO) and ESBL in LTCF, including transmission patterns, rates of infection, and factors associated with infections. It may be necessary to extend the requirements and precautions to MDRO and ESBL-producers.

Extensive Drug Resistance of Strong Biofilm-Producing Acinetobacter baumannii Strains Isolated from Infections and Colonization Hospitalized Patients in Southern Poland.

Acinetobacter baumannii (AB) is a bacterium that causes infections, particularly in immunocompromised patients. Treatment is challenging due to biofilm formation by AB strains, which hinders antibiotic effectiveness and promotes drug resistance. The aim of our study was to analyze the biofilm-producing capacity of AB isolates from various forms of infections in relation to biofilm-related genes and their drug resistance. We tested one hundred isolates for biofilm formation using the crystal violet microplate method. Drug resistance analyses were performed based on EUCAST and CLSI guidelines, and biofilm genes were detected using PCR. All tested strains were found to form biofilms, with 50% being ICU strains and 72% classified as strong biofilm producers. Among these, 87% were extensively drug-resistant (XDR) and 2% were extra-extensively drug-resistant (E-XDR). The most common gene set was bap, bfmS, csuE, and ompA, found in 57% of all isolates. Our research shows that, regardless of the form of infection, biofilm-forming strains can be expected among AB isolates. The emergence of E-XDR and XDR strains among non-ICU infections highlights the necessity for the rational use of antibiotics to stop or limit the further acquisition of drug resistance by A. baumannii.

Oral Microbiota-One Habitat or Diverse Niches? A Pilot Study of Sampling and Identification of Oral Bacterial and Fungal Biota in Patients with Type I Diabetes Mellitus Treated with Insulin Pump.

OBJECTIVE: The oral microbiota is a very complex and dynamic microbial ecosystem. Alterations of its balance can result in oral and systemic diseases. We aimed to characterize the microbiota in particular niches of the oral cavity in adult type 1 diabetes patients treated with continuous infusion of insulin with insulin pump (IP). In addition, we aimed to determine optimal sites of oral microbiota sampling in studies of large research groups of patients with DM I. DESIGN: In this pilot study, we sampled the buccal and soft palate mucosa, tongue, palatal and buccal dental surfaces and gingival pockets of adult DM I patients treated with IP. RESULTS: In total, 23 patients were recruited. The oral microbiota was dominated by Streptococus and Neisseria, with a low incidence of cariogenic S. mutans and Lactobacillus, as well as periodontal pathogens such as Prevotella. There were significant differences in overall CFU counts of all strains, Gram-positive, Staphylococci, Streptococci and S. oralis strains between mucosal and dental surface sites. The overall CFU counts of all strains and Gram-positive strains were higher in dental sites vs. mucosal sites (both p < 0.001). CFU counts of S. oralis were significantly higher in dental sites vs. gingival pocket sites (p = 0.013). Candida species were rare. The mucosal sites on the buccae presented lower diversity and bacterial counts. CONCLUSIONS: In the study group of adult DM I patients treated with IP, the microbiota in particular niches of the oral cavity was significantly different. Three distinct and optimally appropriate sampling sites for oral microflora were identified: buccal and palatal mucosa, dental surface and gingival pockets. The results of this study may be the basis for further studies of large groups of patients with DM I.

Oral Myco- and Bacteriobiota and Yeast Infections in Mechanically Ventilated COVID-19 Patients.

Critically ill COVID-19 patients requiring mechanical ventilation in the intensive care unit are at risk of developing invasive candidiasis. In this study we aimed to (1) characterize oral cultivable mycobiota of mechanically ventilated adult COVID-19 patients in an ICU setting by sampling four distinct oral niches in two fixed time points with regards to oral health status, (2) investigate Candida spp. infections in this population, and (3) compare oral mycobiota with selected bacteriobiota strains during the observation in the ICU. We recruited 56 adult COVID-19 patients who qualified for mechanical ventilation. Patients received either standard or extended oral care procedures with tooth brushing. Oral samples were taken first within 36 h and after 7 days of intubation. Yeast-like fungi were identified by MALDI/TOF mass spectrometry. Yeast infection cases were retrospectively analyzed. Candida spp. in oral sampling was identified in 80.4% and 75.7%, C. albicans in 57.1% and 61.1%, and non-albicans Candida species in 48.2% and 47.2% patients at baseline and follow-up, respectively. There were no differences in the overall CFU counts of Candida spp. species and individual Candida species in oral samples, both at baseline and follow-up. At baseline, a higher prevalence of Candida spp. was associated with a higher identification rate of Lactobacillus spp. (64.4% vs. 27.3%, p = 0.041). At follow-up, there was a borderline lower prevalence of Candida spp. in patients with Lactobacillus spp. identified (57.1% vs. 87.0%, p = 0.057). The incidence rate of candidiasis was 5.4% and the incidence density was 3.1/1000 pds. In conclusion, non-albicans Candida species in oral samples were identified in nearly half of patients. Oral health was moderately impaired. A high incidence of yeast infections, including invasive cases, in patients hospitalized in the ICU due to COVID-19 and requiring mechanical ventilation was noted. Severe COVID-19 and disease-specific interventions within the ICU possibly played a major role promoting Candida spp. infections.

Impact of tooth brushing on oral bacteriota and health care-associated infections among ventilated COVID-19 patients: an intervention study.

BACKGROUND: Up to 48% of ventilated coronavirus disease 2019 (COVID-19) patients develop ventilator-associated pneumonia (VAP) during hospitalization in an ICU. Dysbiotic oral microbiota can colonize the lower respiratory tract and lead to VAP. It is recommended to introduce oral care strategies in the ICU to prevent VAP. In this study, we observed the impact of an oral hygienic protocol with tooth brushing on cultivable oral bacteriota, the incidence of HAI and patient safety among mechanically ventilated COVID-19 patients in an ICU setting. METHODS: In this prospective cohort study, we recruited 56 adult COVID-19 patients who qualified for mechanical ventilation. Patients were divided into 2 groups depending on the oral care procedure: standard and extended oral procedures with tooth brushing. Oral bacteriota samples were taken first within 36 h and after 7 days of intubation. Microorganisms were identified by MALDI/TOF mass spectrometry. bacterial health care-associated infection (HAI) cases were retrospectively analyzed by etiology. A PFGE study was performed for Klebsiella pneumoniae to check for clonal spreading of strains from oral bacteriota samples and HAI cases. RESULTS: We observed significant dysbiosis and a decrease in cultivable oral bacteriota diversity, with a high frequency of potentially pathogenic species, including Acinetobacter baumannii and K. pneumoniae. The HAI incidence rate was high (55.2/1000 patient-days), most commonly of K. pneumoniae and A. baumannii etiologies, which correlated with the presence of A. baumannii and K. pneumoniae in the oral samples. Strains isolated from VAP cases were the same as oral isolates in 8 cases. The procedure with tooth brushing led to less frequent identification of A. baumannii in oral samples (55.6% vs. 5.3%, p = 0.001); however, it did not decrease the incidence of HAIs. CONCLUSIONS: Dysbiotic oral bacteriota is an important source of respiratory pathogens. The introduction of tooth brushing in oral hygiene protocols in an ICU setting was effective in decreasing the extent of oral bacteriota dysbiosis; however, it did not reduce the risk of HAIs or mortality. TRIAL REGISTRATION: 1072.6120.333.2020.

Antimicrobial Resistance of Enterobacteriaceae in Bloodstream Infections in Hospitalized Patients in Southern Poland.

Aim: The aim of this study was to highlight antimicrobial resistance among Enterobacteriaceae isolated from bloodstream infections in hospitals in southern Poland. Materials and Methods: The present study includes laboratory-confirmed secondary bloodstream infections (LC-BSIs), in the years 2015-2018, in hospitalized adult patients (≥18). Episodes of BSIs were defined according to the strictly described guidelines. Antimicrobial susceptibility testing was performed with the automated system and the disc diffusion method. Extended-spectrum ß-lactamases (ESBLs)-producing Enterobacteriaceae were detected using the double-disc synergy test. Results: Between 2015 and 2018, 356 episodes of secondary BSIs in 997 patients aged 21-96 years were documented in a prospective study, including 134 (37.6%) ESBL-producing Enterobacteriaceae. Escherichia coli was the predominant pathogen in internal medicine (37.6%) and surgery units (46.8%); in intensive care units (ICUs), Klebsiella pneumoniae was isolated more frequently (33.3%). Enterobacteriaceae were highly resistant to most antimicrobial agents. K. pneumoniae isolates had a higher level of resistance than E. coli, regardless of the unit. Conclusions: The increase in AMR and the widespread distribution of ESBL-producing Enterobacteriaceae in Polish hospitals can be related to the lack of or inappropriate antibiotic treatment.

Cultivable oral bacteriota dysbiosis in mechanically ventilated COVID-19 patients.

Potential interactions between the SARS-CoV-2 virus and the human oral microbiota are currently investigated widely. Patients with COVID-19 requiring mechanical ventilation in an intensive care unit (ICU) setting are at high risk of developing severe complications, including ventilator-associated pneumonia, thus making oral health management important. The aim of this study was to evaluate the oral health status and assess the dysbiosis of cultivable oral bacteriota in COVID-19 patients hospitalized in an ICU with acute respiratory distress within 36 h following intubation. In this prospective cohort study, we recruited 56 adult COVID-19 patients that qualified for mechanical ventilation in the Temporary ICU for COVID-19 Patients of the University Hospital in Krakow. On admission to the ICU, oral health of patients was assessed using the modified Beck Oral Assessment Score (BOAS). Four oral habitats were sampled, namely the buccal mucosa, tongue, buccal dental surface and gingival pocket. Microorganisms were identified by MALDI/TOF mass spectrometry. The mean age of the study population was 66.5 ± 12.7 years, there were 24 (42.9%) females. All patients included in this study were intubated and ventilated in the ICU, with a corresponding high mortality rate (76.8%). On admission to ICU, 76.8% subjects scored 11-20 on the BOAS scale (median 12 [IQR 10-14]), indicating moderate or severe dysfunction of oral health. Potentially pathogenic bacteria were identified in the oral microbiota samples, including Acinetobacter baumannii, Enterococcus faecalis, Escherichia coli and Klebsiella pneumoniae in 23.2%, 39.3%, 17.9%, and 19.6% of patients, respectively. Lactobacillus spp. were present in 57.1% subjects. The mean CFU counts of all bacteria strains in dental brushes were 9.3E+5 (1.4E+6) and in gingival pockets 7.6E+5 (1.4E+6). The highest CFU counts were observed for Enterococcus spp. and, Lactobacillus spp., although these did not differ significantly from CFU counts of Streptococcus spp. and Staphylococcus spp. In this report we comprehensively characterized the oral health condition and cultivable oral bacteriota in COVID-19 patients hospitalized in an ICU with acute respiratory distress within 36 h following intubation. The oral bacteriota showed significant qualitative and quantitative dysbiosis. Hospitalization in an ICU and mechanical ventilation are important factors leading to oral dysbiosis in SARS-CoV-2 patients.

Healthcare-Associated Laboratory-Confirmed Bloodstream Infections-Species Diversity and Resistance Mechanisms, a Four-Year Retrospective Laboratory-Based Study in the South of Poland.

INTRODUCTION: Regardless of the country, advancements in medical care and infection prevention and control of bloodstream infections (BSIs) are an enormous burden of modern medicine. OBJECTIVES: The aim of our study was to describe the epidemiology and drug-resistance of laboratory-confirmed BSI (LC-BSIs) among adult patients of 16 hospitals in the south of Poland. PATIENTS AND METHODS: Data on 4218 LC-BSIs were collected between 2016-2019. The identification of the strains was performed using MALDI-TOF. Resistance mechanisms were investigated according to European Committee on Antimicrobial Susceptibility Testing, EUCAST recommendations. RESULTS: Blood cultures were collected from 8899 patients, and LC-BSIs were confirmed in 47.4%. The prevalence of Gram-positive bacteria was 70.9%, Gram-negative 27.8% and yeast 1.4%. The most frequently isolated genus was Staphylococcus (50% of all LC-BSIs), with a domination of coagulase-negative staphylococci, while Escherichia coli (13.7%) was the most frequent Gram-negative bacterium. Over 4 years, 108 (2.6%) bacteria were isolated only once, including species from the human microbiota as well as environmental and zoonotic microorganisms. The highest methicillin resistant Staphylococcus aureus (MRSA) prevalence was in intensive care units (ICUs) (55.6%) but S. aureus with resistance to macrolides, lincosamides and streptogramins B (MLSB) in surgery was 66.7%. The highest prevalence of E. faecalis with a high-level aminoglycoside resistance (HLAR) mechanism was in ICUs, (84.6%), while E. faecium-HLAR in surgery was 83.3%. All cocci were fully glycopeptide-sensitive. Carbapenem-resistant Gram-negative bacilli were detected only in non-fermentative bacilli group, with prevalence 70% and more. CONCLUSIONS: The BSI microbiology in Polish hospitals was similar to those reported in other studies, but the prevalence of MRSA and enterococci-HLAR was higher than expected, as was the prevalence of carbapenem-resistant non-fermentative bacilli. Modern diagnostic techniques, such as MALDI-TOF, guarantee reliable diagnosis.

Prevalence and Antimicrobial Susceptibility Profiles of Microorganisms Associated with Lower Reproductive Tract Infections in Women from Southern Poland-Retrospective Laboratory-Based Study.

OBJECTIVE: Female infections affecting the genital tract include sexually transmitted diseases, endogenous infections such as vulvovaginal candidiasis, bacterial vaginosis (BV) or aerobic vaginitis (AV) and healthcare-associated infections. The aim of the study was to analyze the etiological factors of the vaginal dysbacteriosis, and the antimicrobial susceptibility of the dominant bacterial and fungal infections in different age groups of outpatient women from the Silesian Region. MATERIALS AND METHODS: A retrospective laboratory-based multi-center study encompassed 4994 women of different ages in Silesian Voivodeship, in the south of Poland; patients who had vaginal swabs collected as per physicians' orders during the period from 1 January 2017 until 30 June 2018 were included in the study. The inclusion criteria were: non-hospitalized female, aged ≤80, with suspected vulvovaginal candidiasis or bacterial vaginosis and clinical sings of infections. RESULTS: Gram-positive cocci were the ones most often isolated: Enterococcus faecalis (29.2%) and Streptoccoccus agalactiae (13.1%), followed by bacilli from the Enterobacteriaceae group, including Escherichia coli (26.3%). The presence of Streptococcus agalactiae was confirmed in 13.1%, slightly more often in the 45-80 age group, and Gardnerella vaginalis in 6.4%, most often in women aged 15-24. The prevalence of yeast-like infections was 24.3%, Candida albicans accounted for 78.3%, whereas among C. non-albicans spp.-C. glabrata dominated (14.9%) followed by C. parapsilosis (3.8%). The highest resistance was observed only in Streptococcus agalactiae as the MLSB mechanism (Macrolide-lincosamide-streptogramin B) was identified in 38.6% of strains. The prevalence of vulvovaginal candidiasis was 24.3%, the highest in women aged 15-44. CONCLUSIONS: Drug resistance in studied vulvovaginitis was associated only with Streptococcus agalactiae. A high proportion of yeast-like aetiology was found, probably associated with recurrent infections. In the analyzed cases only the Amsel criteria and culture methods were used for diagnosis without preparations and microbiological Nugent criteria.

Virulence and Drug-Resistance of Staphylococcus aureus Strains Isolated from Venous Ulcers in Polish Patients.

Infected chronic venous ulcers (VUs) represent a major health problem. We analysed the aerobic microbiome in the VUs, the virulence, and drug-resistance of Staphylococcus aureus (SA) strains. Swabs from 143 outpatients and inpatients Polish subjects were collected. SA strains were tested for drug sensitivity using a phenotyping method and for methicillin-resistant SA (MRSA) and macrolide-lincosamide-streptogramin B (MLSB) resistance using PCR. We analysed virulence genes, the genetic similarity of strains, and performed Staphylococcal cassette chromosome mec typing and Staphylococcal protein A typing. SA was isolated as a single one in 34.9% of cases, 31.5% paired with another pathogen, and 33.6% S. aureus combined with at least two other strains. The majority of SA isolates (68.5%) possessed the virulence lukE gene. Drug resistance was significantly common in hospitalised than in ambulatory patients (OR 3.8; 95%CI 1.8-7.91). MLSB (altogether in 19.6% isolates) were observed mostly in non-hospitalised patients (OR 9.1; 95%CI 1.17-71.02), while MRSA was detected in 11.9% of strains equally. Hospitalisation and patient's age group (aged > 78.0 or < 54.5 years) were significant predictors of the multi-drug resistant SA (MDR-SA). Over 30% of the infected VUs were associated with multi-species biofilms and presence of potentially highly pathogenic microorganisms. Elderly hospitalised patients with chronic venous ulcers are prone to be infected with a MDR-SA.

[Pseudo-outbreak of MRSA in the pediatric units]. / Pseudoepidemia Staphylococcus aureus o fenotypie MRSA na oddzialach pediatrycznych.

The aim of this paper was to investigate the marked increase noted in the number of MRSA isolates recovered from stool of pediatric units patients. The study involved 37 patients with infection of digestive tract and 1 patient with infection of the central nervous system and took place from 2008 to 2009 years. 3 MRSA isolates collected from staff screening were also included. 44 MRSA isolates were identified using the Vitek automated system. Antimicrobial susceptibility patterns were determined by disk-diffusion method. All isolates were typed by PFGE using SmaI enzyme. Enterotoxin A gene was detected by PCR method. The study confirmed horizontal transmission of epidemic strain of MRSA in the pediatric units, but no connection between clinical symptoms and Staphylococcus aureus not produsing the exotoxin excluded a true outbreak. This pseudo-outbreak emphasizes the importance of cooperation between the microbiology lab and ward personnel for both: competent diagnosis of illness as well as nosocomial infection surveillance and control activities. The key to the described situation was failure to use approciate criteria to diagnose infection. On the other hand it turned out to be a good time for preparing new hygiene procedures and an aggressive educational program for ward staff and parents that promotes best transmission prevention practices.

[Occurrence of resistant Enterobacteriaceae isolated from carriers and infections in pediatric wards of a Polish reference hospital]. / Wystepowanie lekoopornych paleczek z rodziny Enterobacteriaceae u dzieci hospitalizowanych na oddzialach pediatrycznych szpitala wysokospecjalistycznego.

Study was carried out from June to November 2008 in the group of newborn babies hospitalized at the highly specialized hospital. Twenty-six clinical isolates of ESBL-positive rods were collected from 24 patients. Infections incidence was confirmed on the level 3.4/1000 patientdays (pds), among infection dominated blood infection (41%) and pneumonia (31%). All isolates were analyzed for the presence of genes of the resistance (bla)SHV, (bla)TEM i (bla)CTX-M by the multiplex PCR amplification. Isolates were genotyped by PFGE. All isolates were characterized by the presence of the (bla)CTX-M gene. The most of ESBL-positive strains were polyclonal. Three endemic clones of the bacteria were distinguished (among Enterobacter and Klebsiella) which could be moved between patients. Appearing of infection among hospitalized newborn babies showed no relation with the frequency isolation of strains with ESBL phenotype during the period of the study at hospital. The dissemination of ESBLs is due to clonal spread or plasmid dissemination among or between species.