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Introduction. Extended-spectrum β-lactamase (ESβL)-producing Klebsiella pneumoniae is currently one of the most common causes of nosocomial infections worldwide. The study aimed to characterize antibiotic resistance profile, the prevalence of selected genes encoding ESβLs, virulence, and the genetic relationship in 139 K. pneumoniae isolates identified in John Paul II Specialist Hospital in Southern Poland, collected in 2016. Materials/Methods. Bacterial identification and the preliminary antibiotic susceptibilities was performed using the VI-TEK® 2 Compact automated system. Genes encoding ESβLs were amplified by CTX-Mplex PCR and PCR reactions. The presence of nine genes encoding virulence factors was studied by multiplex PCR. Clonality was investigated by PFGE after digestion with SpeI endonuclease. Results. K. pneumoniae were mostly recovered from the respiratory tract (40.3%), urine (32.4%), wound swabs (19.4%) and blood (5%). In summary, 82.7% of strains were classified as multidrug resistant (MDR). All isolates were confirmed as ESβL producers and carried bla CTX-M-type (85.6%), bla SHV (82%), bla TEM (77.7%), bla CTX-M-9 (75.6%) and bla CTX-M-1 (1.4%) in various combinations. Moreover, triple bla genes were observed in 72% of isolates. The most common virulence-as-sociated genes found among the isolates were entB (91.4%), ybtS (55.4%), iutA (55.4%), magA (53.2%), kfu (14.4%), K2 (11.5%), mrkD (10.1%), rmpA (7.9%) and allS (5%). The PFGE analysis identified 4 major clusters (A–D) comprising 61% of the entire collection. Conclusions. Our results indicate that the presence of a wide variety of MDR K. pneumoniae harbor ESβLs and virulence genes. Studies on molecular epidemiology of ESβL-producing K. pneumoniae isolates are needed, particularly for epidemiological surveillance in the hospital environment.

The need for mass population vaccination against Covid-19 poses a public health problem. Allergic symptoms occurring after the 1st dose of the vaccine may result in resignation from the administration of the 2nd dose. However, the majority of patients with mild and/or non-immediate symptoms may be safely vaccinated. The only absolute contraindication to administration of the vaccine is an anaphylactic reaction to any of its ingredients. Polyethylene glycol (PEG), widely used as an excipient in various vaccines, is considered the primary cause of allergic reactions associated with administration of Comirnaty (Pfizer/BioNTech) and Covid-19 Vaccine (Moderna) vaccines. However, hypersensitivity to PEG reported to date seems very rare, considering its widespread use in multiple everyday products, including medicines and cosmetics. In the paper, current literature data describing mechanisms of hypersensitivity reactions to PEG, their clinical symptoms and diagnostic capabilities are presented. Undoubtedly, the issue of hypersensitivity to PEG warrants further research, while patients with the diagnosis require individual diagnostic and therapeutic approach.

Hereditary angioedema (HAE) due to C1-inhibitor (C1-INH) deficiency is a rare disease characterized by recurrent swellings. This study aims to determine (i) the clinical characteristics of the HAE patient population from Poland, and (ii) real-life patients’ treatment practices. A cross-sectional study involved 138 adult HAE patients (88 females, 50 males) treated in six regional HAE centers in Poland. Consecutive patients during routine follow-up visits underwent a structured medical interview on the clinical characteristics of the course and treatment of HAE attacks within the last six months. A total of 118 of 138 patients was symptomatic. They reported in total 2835 HAE attacks predominantly peripheral and abdominal, treated with plasma-derived C1-INH (61.4%), icatibant (36.7%) and recombinant C1-INH (1.9%). An amount of 116 patients carried the rescue medication with them while traveling, and 74 patients self-administrated on demand treatment. There were twice as many symptomatic women (n = 78) as there were men (n = 40). Women treated their HAE attacks significantly more often than men. Older patients (≥65 years) reported a longer delay in diagnosis, and practiced the self-administration of rescue medication less frequently in comparison to other patients. Clinical features of the surveyed population are similar to other European, but not Asian, HAE patient groups. Self-administration still remains an unmet medical need. Some distinct HAE patients may require special attention due to the severe course of the disease (females) or a delay in diagnosis (the elderly).

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.

Wstęp: W wielu dotychczas przeprowadzonych badaniach oceniano związek między cukrzycą a mikroflorą jelitową. Obserwowano zmianę mikroflory jelitowej pod wpływem inhibitorów dipeptydylopeptydazy-4 w modelach zwierzęcych. Celem niniejszego badania była ocena wpływu linagliptyny na florę bakteryjną okrężnicy u ludzi. Materiał i metody: W prospektywnym badaniu kohortowym wzięło udział 24 pacjentów: 5 chorych na cukrzycę monogenową, związaną z mutacją HNF1A, i 19 chorych na cukrzycę typu 2. Próbki kału pobrano przed i 4 tygodnie po intensyfikacji aktualnego leczenia za pomocą linagliptyny lub pochodnej sulfonylomocznika. Z próbek kału wyizolowano rRNA, a następnie wykonano sekwencjonowanie 16S nowej generacji. Wyniki: U 9 pacjentów do leczenia dołączono linagliptynę, a u 15 pacjentów dołączono sulfonylomocznik lub zwiększono jego dawkę. Po leczeniu linagliptyną nie zaobserwowano zmian na poziomach taksonomicznych L2–L7 na podstawie analizy składu mikrobiomów (ANCOM). To samo odnosiło się do różnorodności alfa [wskaźnik różnorodności Shannona, p = 0,59; wskaźnik równomierności Pielou, p = 0,68; obserwowane operacyjne jednostki taksonomiczne (OTU), p = 0,77] i różnorodności beta (nieważony wskaźnik UniFrac, p = 0,99; ważony wskaźnik UniFrac, p = 0,93; wskaźnik Braya–Curtisa, p = 0,98; wskaźnik Jaccarda, p = 0,99). Również po intensyfikacji leczenia pochodną sulfonylomocznika nie zaobserwowano zmian na poziomach taksonomicznych L2–L7 w ANCOM ani zmian w różnorodności alfa (wskaźnik różnorodności Shannona, p = 0,19; wskaźnik równomierności Pielou, p = 0,21; obserwowane OTU, p = 0,42) i różnorodności beta (nieważony wskaźnik UniFrac, p = 0,99; ważony wskaźnik UniFrac, p = 0,99; wskaźnik Braya–Curtisa, p = 1; wskaźnik Jaccarda, p = 0,99). Wnioski: Po 4 tygodniach od dołączenia linagliptyny do aktualnego leczenia cukrzycy nie zaobserwowano zmian w mikroflorze okrężnicy. Konieczne są dalsze badania w celu ustalenia, czy linagliptyna wpływa na mikroflorę okrężnicy u ludzi. Introduction. Many studies have evaluated the relationship between diabetes and microbiota. In animal models, the dipeptidyl peptidase-4 inhibitors altered the gut microbiota. We investigated whether linagliptin alters the gastrointestinal flora in humans. Materials and methods. This prospective cohort study enrolled 24 patients: 5 patients with maturity onset diabetes of the young associated with HNF1A mutation and 19 patients with type 2 diabetes mellitus. Stool samples were collected at baseline and 4 weeks after treatment intensification with either linagliptin or a sulphonylurea alongside current treatment. Faecal 16S rRNA was analysed by next-generation sequencing. Results. Nine patients initiated linagliptin whereas 15 patients initiated or increased the dose of a sulphonylurea. After linagliptin treatment, we did not observe changes in taxa in L2–L7 based on analysis of composition of microbiomes (ANCOM). The same held true for pairwise alpha diversity (Shannon diversity, p = 0.59; Pielou’s measure of evenness, p = 0.68; and observed operational taxonomic units [OTUs], p = 0.77) and beta diversity distances (unweighted UniFrac, p = 0.99; weighted UniFrac, p = 0.93; Bray-Curtis, p = 0.98; and Jaccard, p = 0.99). Similarly, after sulphonylurea intensification, we did not observe changes in taxa in L2–L7 in ANCOM, nor were there changes in alpha diversity (Shannon diversity, p = 0.19; Pielou’s measure of evenness, p = 0.21; and observed OTUs, p = 0.42) or beta diversity distances (unweighted UniFrac, p = 0.99; weighted UniFrac, p = 0.99; Bray-Curtis, p = 1; and Jaccard, p = 0.99). Conclusion. We did not observe changes in colonic microbiota 4 weeks after addition of linagliptin to current diabetes treatment. Further studies are required to determine whether linagliptin influences the colonic microbiota in humans

Until now, the simultaneous influence of air pollution assessed by measuring the objective marker of exposition (1-hydroxypirene, 1-OHP) and atopy on the development of allergic airway diseases has not been studied. The aim of this study was to determine the pathomechanism of the allergic response to PM2.5 in atopic and non-atopic patients. We investigated the changes in peripheral blood basophil activity of patients after stimulation with the birch pollen allergen alone, the allergen combined with PM2.5 (BP), PM2.5 alone, a concentration of 1-OHP in urine, and a distance of residence from the main road in 30 persons. Activation by dust alone was positive for all concentrations in 83% of atopic and 75% of non-atopic assays. In the group of people with atopy, the simultaneous activation of BP gave a higher percentage of active basophils compared to the sum of activation with dust and birch pollen alone (B + P) for all concentrations. The difference between BP and B + P was 117.5 (p = 0.02) at a PM concentration of 100 μg. Such a relationship was not observed in the control group. The correlation coefficient between the distance of residence from major roads and urinary 1-OHP was 0.62. A Pearson correlation analysis of quantitative variables was performed, and positive correlation results were obtained in the atopy group between BP and 1-OH-P. Exposure to birch pollen and PM2.5 has a synergistic effect in sensitized individuals. The higher the exposure to pollutants, the higher the synergistic basophil response to the allergen and PM in atopic patients.

The studies on microbiome in the human digestive tract indicate that fungi could also be one of the external factors affecting development of diabetes. The aim of this study was to evaluate the quantitative and qualitative mycobiome composition in the colon of the adults with type 1 (T1D), n = 26 and type 2 (T2D) diabetes, n = 24 compared to the control group, n = 26. The gut mycobiome was characterized in the stool samples using the analysis of the whole internal transcribed spacer (ITS) region of the fungal rDNA gene cluster by next-generation sequencing (NGS) with increased sensitivity. At the L2 (phylum) level, Basidiomycota fungi were predominant in all 3 study groups. Group T1D presented significantly lower number of Ascomycota compared to the T2D group, and at the L6 (genus) level, the T1D group presented significantly lower number of Saccharomyces genus compared to control and T2D groups. In the T1D group, a significant positive correlation between total cholesterol and low-density lipoprotein cholesterol (LDL-C) levels and fungi of the genus Saccharomyces, and in the T2D group, a negative correlation between the total cholesterol level and Malassezia genus was found. The obtained results seem to be a good foundation to extend the analysis of the relationship between individual genera and species of fungi and the parameters determining the metabolism of carbohydrates and lipids in the human body.

Background/Objectives: The genus Acinetobacter demonstrates resistance to antibiotics and has been shown to spread in the hospital environment causing epidemic outbreaks among hospitalized patients. The objectives of the present study was to investigate the antibiotic resistance, biofilm formation, and clonality among Acinetobacter baumannii strains. Materials and Methods: The study involved 6 (I Outbreak) and 3 (II Outbreak) A. baumannii strains isolated from patients hospitalized in vascular surgery unit. Results: All tested A. baumannii strains were extensively drug resistant (XDR) and all the isolates were carbapenem-resistant and among them, all carried the blaOXA-51 gene, the blaOXA-24 gene, as well as the blaOXA-23 gene. All of the investigated strains had the ability to form a biofilm, but all of them produced less biofilm than the reference strain. Multi-locus sequence typing (MLST) showed that all strains belonged to the ST2 clone. Pulsed-field gel electrophoresis (PFGE) divided the tested outbreak strains into two clones (A and B). Conclusion: This study shows a nosocomial spread of XDR A. baumannii ST2 having the blaOXA-51 gene, the blaOXA-24 gene, as well as the blaOXA-23 gene, low biofilm formers, that was prevalent in the vascular surgery unit. To identify the current situation of vascular surgery departments targeted epidemiological investigation was needed. Effective implementation of infection control prevented the spread of the epidemic outbreaks.

Recently, several studies explored associations between type 1 diabetes (T1DM) and microbiota. The aim of our study was to assess the colonic microbiota structure according to the metabolic control in T1DM patients treated with insulin pumps. We studied 89 T1DM patients (50.6% women) at the median age of 25 (IQR, 22&ndash, 29) years. Pielou&rsquo, s evenness (p = 0.02), and Shannon&rsquo, s (p = 0.04) and Simpson&rsquo, s diversity indexes (p = 0.01), were higher in patients with glycosylated hemoglobin (HbA1c) &ge, 53 mmol/mol (7%). There were no differences in beta diversity between groups. A linear discriminant analysis effect size (LEfSe) algorithm showed that one family (Ruminococcaceae) was enriched in patients with HbA1c &lt, 53 mmol/mol, whereas one family (Streptococcaceae) and four species (Ruminococcus torques, unclassified species of Lactococcus, Eubacteroim dolichum, and Coprobacillus cateniformis) were enriched in patients with HbA1c &ge, 53 mmol/mol. We found that at class level, the following pathways according to Kyoto Encyclopedia of Genes and Genomes were enriched in patients with HbA1c &lt, 53 mmol/mol: bacterial motility proteins, secretion system, bacterial secretion system, ribosome biogenesis, translation proteins, and lipid biosynthesis, whereas in patients with HbA1c &ge, 53 mmol/mol, the galactose metabolism, oxidative phosphorylation, phosphotransferase system, fructose, and mannose metabolism were enriched. Observed differences in alpha diversity, metabolic pathways, and associations between bacteria and HbA1c in colonic flora need further investigation.

Eight aerosol samples were collected in Krakow using a low-volume sampler in February and March 2019 during variable meteorological conditions and times of the day, to study their single particles’ properties (size, morphology and chemical composition analyzed using a scanning electron microscope fitted with an energy-dispersive spectrometer) and microbiological characteristics. The content of particles of different chemical compositions larger than 2.5 μm was low. Considering the number of the particles, submicron particles strongly dominated with a high content of ultrafine particles (nanoparticles). Tar ball-type particles were relatively common in the studied samples, while soot was the dominant component. Soot was present as small agglomerates composed of few particles, but also as bigger agglomerates. Metal-containing particles of various chemical characteristics were abundant, with transition metals commonly occurring in these particles. The physicochemical characteristics of aerosols indicate that despite a relatively low mass concentration, their adverse health impact could be very strong because of the high content of nanoparticles, the abundance of soot and other fuel combustion-related particles, and the high incidence of transition metal-rich particles. Microbiological analysis was based on cultures on both solid and liquid agar. The MALDI-TOF method was used for species identification—for bacteria and fungi. Twelve different species of bacteria were isolated from the collected samples of aerosols. The most frequently isolated species was Gram-positive sporulating Bacillus licheniformis. The isolated mold fungi were of the genus Aspergillus.

Background/Objectives: The genus Acinetobacter demonstrates resistance to antibiotics and has been shown to spread in the hospital environment causing epidemic outbreaks among hospitalized patients. The objectives of the present study was to investigate the antibiotic resistance, biofilm formation, and clonality among Acinetobacter baumannii strains. Materials and Methods: The study involved 6 (I Outbreak) and 3 (II Outbreak) A. baumannii strains isolated from patients hospitalized in vascular surgery unit. Results: All tested A. baumannii strains were extensively drug resistant (XDR) and all the isolates were carbapenem-resistant and among them, all carried the blaOXA-51 gene, the blaOXA-24 gene, as well as the blaOXA-23 gene. All of the investigated strains had the ability to form a biofilm, but all of them produced less biofilm than the reference strain. Multi-locus sequence typing (MLST) showed that all strains belonged to the ST2 clone. Pulsed-field gel electrophoresis (PFGE) divided the tested outbreak strains into two clones (A and B). Conclusion: This study shows a nosocomial spread of XDR A. baumannii ST2 having the blaOXA-51 gene, the blaOXA-24 gene, as well as the blaOXA-23 gene, low biofilm formers, that was prevalent in the vascular surgery unit. To identify the current situation of vascular surgery departments targeted epidemiological investigation was needed. Effective implementation of infection control prevented the spread of the epidemic outbreaks.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an important bacterium of nosocomial infections. In this study, CRKP strains, which were mainly isolated from fecal samples of 14 patients in three wards of the hospital in the Silesia Voivodship, rapidly increased from February to August 2018. Therefore, we conducted microbiological and molecular studies of the CRKP isolates analyzed. Colonized patients had critical underlying diseases and comorbidities; one developed bloodstream infection, and five died (33.3%). Antibiotic susceptibilities were determined by the E-test method. A disc synergy test confirmed carbapenemase production. CTX-Mplex PCR evaluated the presence of resistance genes blaCTX-M-type, blaCTX-M-1, blaCTX-M-9, and the genes blaSHV, blaTEM, blaKPC-2, blaNDM-1, blaOXA-48, blaIMP, and blaVIM-1 was detected with the PCR method. Clonality was evaluated by Multi Locus Sequence Typing (MLST) and Pulsed Field Gel Electrophoresis (PFGE). Six (40%) strains were of XDR (Extensively Drug-Resistant) phenotype, and nine (60%) of the isolates exhibited MDR (Multidrug-Resistant) phenotype. The range of carbapenem minimal inhibitory concentrations (MICs, μg/mL) was as follows doripenem (16 to >32), ertapenem (> 32), imipenem (4 to > 32), and meropenem (> 32). PCR and sequencing confirmed the blaCTX-M-15, blaKPC-2, blaOXA-48, and blaVIM-1 genes in all strains. The isolates formed one large PFGE cluster (clone A). MLST assigned them to the emerging high-risk clone of ST147 (CC147) pandemic lineage harboring the blaOXA-48 gene. This study showed that the K. pneumoniae isolates detected in the multi-profile medical centre in Katowice represented a single strain of the microorganism spreading in the hospital environment.

Introduction: The paper presents the phenotypic and molecular characteristics of the collection of 18 P. aeruginosa clinical isolates used as host indicators to study the lytic range of 12 phages against P. aeruginosa. Methods: The phages host ranges were assayed by spot tests. Phenotypic characteristics of strains was investigated by the API 20NE biochemical fingerprinting, oxidase tests, the production of pyocyanin, fluorescein and L-arginine dihydrolase. Resistance profiles were analyzed. The PCR method and sequencing were used to study the distribution the genes of alkaline protease (aprA), exotoxin A (exoA), elastase B (lasB), exotoxins (exoS/T/U/Y), phenazine modyfing genes (phzM, phzS) and to identify selected β-lactamases (blaGES, blaIMP, blaKPC, blaOXA-2, blaOXA-10, blaPER, blaSPM-1, blaSHV, blaTEM, blaVIM). Additionaly, the genetic diversity was investigated by PFGE.116 M. Brzychczy-Włoch i inni Nr 2-4 Results: Twenteen newly isolated P. aeruginosa phages were found to lyse 100% of the analyzed strains. Phages PAR_3 and PAR_10 exhibited the highest lytic activity against isolates, lysing, 77,8% strains tested. The other phages, PAR_9 and PAR_12, presented generally weaker activity against bacteria, lysing respectively, 50% and 44,4% of tested strains. AprA, exoA, phzM, phzS were presented in all strains; lasB in 77,8%. The most frequentlycombination of egzoenzyme genes S+/T+/U-/Y+ in 78% isolates was remarked. In collection, 18 different resistance profiles were observed and 44% isolates were classified as MDR. The blaGES was the most prevalent gene (44%), followed by blaSPM-1 and blaTEM detected in 17% and 11,1% isolates, respectively. BlaOXA-2 was detected in only 5,5% of all isolates. In PFGE method, 18 singletons (A-S) were identified. No relationship between resistance, virulence and PFGE groups was found. Conclusion: In summarize, all phages infect multiple host species and showed a broad lytic spectrum. All bacteria tested were infected by multiple phages and displayed a wide range of susceptibility. In general, we observed a high degree of genetic diversity and individuality of the studied P. aeruginosa collection, bacteriophage hosts.

Numerous scientific studies confirm that, apart from environmental and genetic factors, a significant role is played by gastrointestinal microbiota in the aetiology of type 2 diabetes and obesity. Currently, scientists mainly focus on the distal intestinal microbiota, while the equally important proximal parts of the intestine are overlooked. The aim of the study was a qualitative analysis of the structure of the duodenal mucosa microbiota in groups of patients with obesity and with type 2 diabetes and where obesity qualified for bariatric surgery: sleeve gastrectomy. The microbiological results obtained were compared with some clinical parameters. As a result, it was possible to determine the microbiological core that the treatment and control groups had in common, including phyla: Firmicutes, Proteobacteria, and Actinobacteria. The patients with obesity and with type 2 diabetes and obesity presented a significantly lower number of genus Bifidobacterium compared to healthy subjects. Furthermore, the numbers of Bifidobacterium were positively correlated with the high density lipoprotein (HDL) concentration in the groups under study. The obtained results indicate that bacteria of the genus Bifidobacterium should be considered in the future in the context of a potential biomarker in the progress of type 2 diabetes and obesity.

This study investigated differences in perceptions of hand hygiene and protective glove use among patients and health care workers (HCWs) in Poland. We conducted a survey using an original questionnaire among 462 respondents, including 173 (37.4%) patients and 289 (62.6%) HCWs; HCWs demonstrated poor familiarity with the My 5 Moments for Hand Hygiene. The role of protective gloves in preventing health care-associated infection was overestimated by both patients and HCWs.

This study conducts a comparative phenotypic and genetic analysis of C. perfringens strains isolated from two patients hospitalized at the same time in 2017 in the surgical ward of the Provincial Specialist Hospital in Wloclawek (Kujawsko-Pomorskie Province) who developed necrotizing soft tissue infections (NSTI). To explain the recurring cases of this infection, a comparative analysis was performed for these strains and the ones originating from infections recorded at the same hospital in three patients with gas gangrene in 2015. The two C. perfringens isolates studied in 2017 (8554/M/17 from patient No. 1 and 8567/M/17 from patient No. 2) had identical biochemical profiles. A comparison of research results using multiplex PCR from 2017 with a genetic analysis of strains from 2015 enabled us to demonstrate that the strains currently studied have the genes encoding the same toxins (α and β2) as the two strains analyzed in 2015: no. 7143 (patient No. 3) and no. 7149 (patient No. 2). A comparative analysis of the strain profiles obtained with pulsed-field gel electrophoresis (PFGE) in 2017 with the results from 2015 has found one identical and genetically unique restriction profile, corresponding to one clone of C. perfringens comprising of two strains: no. 8567/M/17 (patient No. 2 in 2017) and no. 7143 (patient No. 3 in 2015). The epidemiological data and detailed analysis of the course of both events suggest that this clone of C. perfringens possibly survived in adverse conditions of the external environment in the operating block of this hospital for many months. This study conducts a comparative phenotypic and genetic analysis of C. perfringens strains isolated from two patients hospitalized at the same time in 2017 in the surgical ward of the Provincial Specialist Hospital in Wloclawek (Kujawsko-Pomorskie Province) who developed necrotizing soft tissue infections (NSTI). To explain the recurring cases of this infection, a comparative analysis was performed for these strains and the ones originating from infections recorded at the same hospital in three patients with gas gangrene in 2015. The two C. perfringens isolates studied in 2017 (8554/M/17 from patient No. 1 and 8567/M/17 from patient No. 2) had identical biochemical profiles. A comparison of research results using multiplex PCR from 2017 with a genetic analysis of strains from 2015 enabled us to demonstrate that the strains currently studied have the genes encoding the same toxins (α and β2) as the two strains analyzed in 2015: no. 7143 (patient No. 3) and no. 7149 (patient No. 2). A comparative analysis of the strain profiles obtained with pulsed-field gel electrophoresis (PFGE) in 2017 with the results from 2015 has found one identical and genetically unique restriction profile, corresponding to one clone of C. perfringens comprising of two strains: no. 8567/M/17 (patient No. 2 in 2017) and no. 7143 (patient No. 3 in 2015). The epidemiological data and detailed analysis of the course of both events suggest that this clone of C. perfringens possibly survived in adverse conditions of the external environment in the operating block of this hospital for many months.

Anaphylaxis is an increasing problem in public health. The food allergens (mainly milk, eggs, and peanuts) are the most frequent cause of anaphylaxis in children and youth. In order to define the cause of anaphylaxis, skin tests, the determination of the concentration of specific IgE in the blood and basophil activation test are conducted. In vitro tests are preferred due to the risk of allergic response during in vivo tests. Component-resolved diagnosis (CRD) is an additional tool in allergology, recommended in the third level of diagnostics when there are diagnostic doubts after the above mentioned tests have been carried out. The paper presents 3 cases of patients with anaphylactic response, and the application of CRD in these patients helped in planning the treatment. Patient 1 is a 4-year-old boy with diagnosed atopic dermatitis and bronchial asthma reported an anaphylactic shock at the age of seven months caused by cow's milk and the exacerbation of bronchial asthma after eating some fruit. Patient 2 is a 35-year-old woman who has had anaphylactic shock three times: in June 2015, 2016, and 2017 and associates these episodes with the consumption of dumplings with a caramel, bun, and the last episode took place during physical exertion few hours after eating waffle. Patient 3 is a 26-year-old man with one-time loss of consciousness after eating mixed nuts and drinking beer. CRD offers the possibility to conduct a detailed diagnostic evaluation of patients with a history of anaphylactic reaction.

Qualification for specific immunotherapy (SIT) according to the guidelines of the European Academy of Allergy and Clinical Immunology (EAACI) includes medical history, skin prik tests (SPT) and/or measuring the concentration of sIgE. It is necessary to perform additional diagnostic tests in case of discrepancies between the history and the results of SPT/sIgE or differences between SPT and sIgE. Basophil activation test (BAT) assesses the expression of activation markers of these cells, eg. CD63 and CD203c after stimulation. The aim of our study was to evaluate the usefulness of BAT in the qualification for the SIT in comparison to the SPT and sIgE and in case of discrepancies between the results of SPT and sIgE. The study included 30 patients with allergic rhinitis (AR) caused by allergy to house dust mite (Dermatophagoides pteronyssinus, Dp) or birch pollen qualified for SIT. All patients had SPT, sIgE and BAT determination. The group of patients with allergy to birch was a control group for Dp allergic and vice versa. BAT with CD63 antigen expression was performed using a Flow2CAST test. Basophils were stimulated with allergen preparation (50, 500, and 5000 SBU/ml concentrations). BAT results were expressed as a stimulation index (SI). For optimal concentrations of 50 and 500 SBU/ml parameters comparing BAT to SPT and sIgE as the gold standards were consecutively: sensitivity 82-100% and 93-100%, specificity 50-94% and 47-89%, positive predictive value 65- 94% and 61-87%, negative predictive value 86-100% and 93-100%. Correlation BAT - SPT and BAT - sIgE ranged within 0.59 to 0.84 and 0.51 to 0.72. BAT was helpful in 2 of 30 patients with incompatible results of SPT and sIgE. Optimal concentrations for basophil stimulation are 50 and 500 SBU/ ml. BAT may be useful diagnostic tool in the qualification for the SIT in case of discrepancies between the results of SPT and sIgE.

Nosocomial bloodstream infections (BSI) among patients in intensive care units (ICU) are the most common form of nosocomial infections and they are serious complications of the treatment process. The etiological factors of these infections and the proper antibiotic therapy can influence the effects of patient treatment. The aim of the study was to analyze the epidemiology, microbiology and antibiotic therapy of nosocomial BSI in Polish ICU patients.The retrospective study covered patients hospitalized in seven ICUs for adults with a general profile, located in southern Poland. The studied units participated in a multicentre standardized infection control program in 2013-2015. 2,547 patients hospitalized more than 48 hours in ICUs were qualified to the study. The study was conducted in accordance with the methodology recommended by the European Center for Disease Prevention and Control (ECDC) as part of the European Healthcare-Associated Infections Surveillance Network (HAI-Net): protocols and the criteria for BSI recognition were standardized.The incidence of BSI was 7.2% per 100 admissions and the incidence density was 9.2 per 1000 persondays of hospitalization. Among 184 BSI cases, 65 cases (35.3%) were primary BSI; 87 cases were secondary BSI (47.3%) (related to an infection occurring elsewhere in the patient’s body), the remaining 32 cases (17.4%) were classified as BSI of unknown origin. Mortality was 20%. The dominant etiological factors in BSI related to central venous catheter were coagulase-negative staphylococci in 20.8% of cases, but in secondary BSI it was Acinetobacter baumannii in 34.5% of cases. Klebsiella pneumoniae showed resistance to third-generation cephalosporins in 96.0% of cases; Acinetobacter baumannii showed resistance to imipenem in 78.8% of cases. Other beta-lactam antibiotics J01D (31.0%) and other antibiotics J01X (26.6%) were most commonly used in the treatment.The BSI incidence in the studied Polish ICUs was twice as high as in other European countries, and secondary BSI were also more frequent – especially secondary to urinary tract infections. The obtained results indicate the necessity of intervention not only in BSI prevention, but also in the field of prevention of urinary tract infections. In addition, microbial etiology and drug resistance indicate the need for urgent actions for their prevention.

Vancomycin-resistant Enterococcus faecium (VREfm) strains have become an important hospital pathogen due to their rapid spread, high mortality rate associated with infections and limited therapeutic options. Vancomycin resistance is predominantly mediated by VanA or VanB phenotypes, which differ as regards maintaining sensitivity to teicoplanin in the VanB phenotype. The majority of VREfm cases in the United States, Europe, Korea, South America and Africa are currently caused by the VanA phenotype. However, the epidemics in Australia and Singapore are chiefly brought about by the VanB phenotype. The rate of VREfm isolate spread varies greatly. The greatest percentage of VREfm is now recorded in the USA, Ireland and Australia. Supervision of VRE is implemented to varying degrees. Therefore, the epidemiological situation in some countries is difficult to assess due to limited data or lack thereof.

Introduction Scientific data indicate a possible influence of gut microbiota on the development of type 1 and type 2 diabetes mellitus (T1DM and T2DM, respectively). Sequence analysis of 16S ribosomal RNA identified several hundred bacterial species of the intestinal ecosystem, most of which cannot be cultured. Objectives We aimed to evaluate gut microbiota composition in adult patients with T1DM and T2DM and establish a link between microbiological test results and patients' clinical data. Patients and methods We examined DNA isolated from fecal samples in 3 groups: healthy volunteers (n = 23), patients with T1DM (n = 22), and patients with T2DM (n = 23). Next‑generation sequencing was performed on the MiSeq platform. Results At the phylum level, the Firmicutes bacteria prevailed (77%) in all groups. At the taxonomic levels L2 (phylum) and L6 (genus), significant differences were demonstrated in bacterial profiles, particularly in the T2DM group. A negative correlation was observed between several genera of bacteria and the percentage of glycated hemoglobin A1c in the T2DM group, while a positive correlation was revealed between bacteria belonging to the genus Bifidobacterium and high‑density lipoprotein cholesterol levels in both T1DM and T2DM groups. Conclusions Our results provide grounds for conducting research in the field of gut microbiota in order to develop individualized therapy for patients with diabetes based on modifying the microbiota composition, as a new method for controlling glycemia. Next‑generation sequencing allows a rapid identification of the DNA of all bacteria present in the sample and their taxonomic classification.

Objectives To evaluate data on outpatient antibiotic use in women post-labour as a potential method of monitoring infections in this group of patients. Methods Demographic and antibiotic prescription data originated from the registries of the National Health Fund (pol. Narodowy Fundusz Zdrowia). The measure of antibiotic use in this study was the percentage of women who purchased the drugs from prescriptions and DDDs. Results Among 67917 females who gave birth in the years 2013-14, 5050 (7.4%) purchased antibiotics prescribed by the obstetrician only. The average number of antibiotics bought per person was equivalent to ∼14 DDDs; in most cases (95.7%) these were β-lactams. Antibiotic use occurred significantly more frequently among younger patients (11.5% patients

Despite improvements in surgical management, sternal wound infection (SWI) following cardiac surgery remains a significant complication.To evaluate pathogens involved in SWI following median sternotomy.We enrolled 164 patients who experienced prolonged sternal wound healing following open-heart surgery. The inclusion criteria were as follows: prolonged sternal wound healing following cardiac surgery via median sternotomy and complete results of microbiological culture obtained from the sternal swab. The exclusion criteria were as follows: partial sternotomy, patients with mechanical sternum dehiscence and incomplete clinical data. Swabs provided information on the type of microorganism present in the wound and the susceptibility of the microorganism to specific antibiotics.One hundred and fourteen (69.5%) patients aged 68 (interquartile range: 60-76) years developed SWI with positive culture. The in-hospital mortality rate was 21.0% in this group. The most common pathogens includedMicroorganisms associated with SWI in our study were mainly commensals, with the most common pathogen beingPomimo udoskonaleń w procedurach chirurgicznych nadal głównym powikłaniem po operacjach kardiochirurgicznych są zakażenia ran mostka.Ocena patogenów, które wywołują zakażenia ran mostka po sternotomii pośrodkowej.Do badania włączono 164 pacjentów, u których stwierdzono przedłużone gojenie rany mostka po operacji kardiochirurgicznej. Kryteria włączenia stanowiły przedłużone gojenie się rany mostka po operacji kardiochirurgicznej z dostępu przez sternotomię pośrodkową i kompletne wyniki hodowli mikrobiologicznej uzyskane z wymazu z mostka, natomiast kryteria wyłączenia – częściowa sternotomia, mechaniczne rozejście się mostka i niekompletne dane kliniczne. Mikroorganizmy były identyfikowane za pomocą hodowli mikrobiologicznej, na podstawie której określano także ich lekowrażliwość.U 114 (69,5%) pacjentów w wieku 68 lat (rozstęp międzykwartylowy: 60–76) doszło do zakażenia rany mostka. Śmiertelność w tej grupie wynosiła 21%. Najczęściej izolowane patogeny to:Mikroorganizmy powodujące wyżej wymienione zakażenia to głównie komensale, a najczęściej występującym patogenem był

Copper and its alloys are known for their antimicrobial activity, which makes them appealing materials for various touch surfaces in public facilities. These materials are also known for being prone to tarnishing, especially in contact with human palm sweat. The paper describes investigations on tarnishing of copper and various copper alloys by oxidation at elevated temperatures. After evaluation of thickness and chemical composition of oxide layers, microbiological tests were carried out in order to determine the impact of oxidation on antimicrobial efficiency of copper alloys.

Three Streptococcus agalactiae (group B streptococci, GBS) immunoreactive proteins: enolase (47.4 kDa), inosine 5'-monophosphate dehydrogenase (IMPDH) (53 kDa) and molecular chaperone GroEL (57 kDa) were subjected to investigation. Enolase protein was described in our previous paper, whereas IMPDH and GroEL were presented for the first time. The aim of our paper was to provide mapping of specific epitopes, highly reactive with umbilical cord blood serum. Bioinformatic analyses allowed to select 32 most likely epitopes for enolase, 36 peptides for IMPDH and 41 immunoreactive peptides for molecular chaperone GroEL, which were synthesized by PEPSCAN. Ten peptides: two in enolase, one in IMPDH and seven in molecular chaperone GroEL have been identified as potentially highly selective epitopes that can be used as markers in rapid immunological diagnostic tests or constitute a component of an innovative vaccine against GBS infections.

BACKGROUND In the Malopolska province, the first case of vancomycin resistant enterococci (VRE) occurrence was an outbreak in 2001 caused by strains of the genus E. faecium carrying the vanA operon. OBJECTIVES The aim of this study is to determine the antimicrobial resistance and the occurrence of virulence determinants among Enterococcus spp. in patients hospitalized in the Malopolska region in 2015. MATERIAL AND METHODS Antimicrobial susceptibility was determined by disc diffusion and the E test. The presence of aminoglycoside and glycopeptide resistance genes and virulence genes (asa1, gelE, cylA, esp, hyl) was investigated using multiplex polymerase chain reaction (PCR). Also, the presence of IS16 was investigated. The activity of gelatinase, cytolysin (hemolysin), and DNase was tested. RESULTS All E. faecalis were susceptible to ampicillin, vancomycin, teicoplanin, linezolid and tigecycline. All E. faecium strains were susceptible to quinupristin-dalfopristin. Resistance to ampicillin and vancomycin was detected among all E. faecium isolates from hospitals C and D. 87.32% of E. faecium presented high-level aminoglycoside-resistant (HLAR) phenotype, including 78.33% of strains from hospital C and 100% from hospital D. In hospital C (98.3%) and D (96%), resistance to ciprofloxacin, levofloxacin and norfloxacin was observed. Gene esp was detected in all E. faecium isolates and the majority of E. faecium isolates carried hyl (97%). In E. faecalis, different combinations of virulence genes were detected. All analyzed E. faecium strains showed the presence of IS16 insertion element. CONCLUSIONS E. faecalis isolates were more susceptible to antimicrobials than E. faecium, which were largely multidrug-resistant. E. faecalis strains have diverse virulence factors. E. faecium showed a high percentage of hyl and esp genes and the presence of IS16.

The gold standard in microbiological diagnostics of bacteremia is a blood culture in automated systems. This method may take several days and has low sensitivity. New screening methods that could quickly reveal the presence of bacteria would be extremely useful. The objective of this study was to estimate the effectiveness of these methods with respect to blood cultures in the context of antibiotic therapy. Blood samples from 92 children with sepsis were analyzed. Blood cultures were carried out in standard automated systems. Subsequently, FISH (Fluorescent In-Situ Hybridization) and nested multiplex-real-time-PCR (PCR) were performed. Blood cultures, FISH and PCR yielded positive results in 18%, 39.1%, and 71.7% of samples, respectively. Significant differences were found between the results obtained through culture before and after induction of antibiotherapy: 25.5% vs. 9.7%. There was no significant difference in FISH and PCR results in relation to antibiotics. The three methods employed demonstrated significant differences in detecting bacteria effectively. Time to obtain test results for FISH and PCR averaged 4-5 hours. FISH and PCR allow to detect bacteria in blood without prior culture. These methods had high sensitivity for the detection of bacteremia regardless of antibiotherapy. They provide more timely results as compared to automated blood culture, and may be useful as rapid screening tests in sepsis. The gold standard in microbiological diagnostics of bacteremia is a blood culture in automated systems. This method may take several days and has low sensitivity. New screening methods that could quickly reveal the presence of bacteria would be extremely useful. The objective of this study was to estimate the effectiveness of these methods with respect to blood cultures in the context of antibiotic therapy. Blood samples from 92 children with sepsis were analyzed. Blood cultures were carried out in standard automated systems. Subsequently, FISH (Fluorescent In-Situ Hybridization) and nested multiplex-real-time-PCR (PCR) were performed. Blood cultures, FISH and PCR yielded positive results in 18%, 39.1%, and 71.7% of samples, respectively. Significant differences were found between the results obtained through culture before and after induction of antibiotherapy: 25.5% vs. 9.7%. There was no significant difference in FISH and PCR results in relation to antibiotics. The three methods employed demonstrated significant differences in detecting bacteria effectively. Time to obtain test results for FISH and PCR averaged 4–5 hours. FISH and PCR allow to detect bacteria in blood without prior culture. These methods had high sensitivity for the detection of bacteremia regardless of antibiotherapy. They provide more timely results as compared to automated blood culture, and may be useful as rapid screening tests in sepsis.