Changes of urine isolates of Escherichia coli and Klebsiella pneumoniae biofilm affect monocytes' response.

The Gram negative rods as Escherichia coli and Klebsiella pneumoniae belong to the most common etiology agents of urinary tract infections. The aim of our study was to assess the diversity of biofilm formed in different urinary tract diseases and their impact on monocytes' adherence and activation. The bacteria were obtained from patients with different kidney problems. Some of the patients were after renal transplantation, some of them were not. Changes in the size and granularity of monocytes, as well as their adherence to biofilm, were assessed using FACSVerse flow cytometer after 1 h co-incubation of monocytes and bacterial biofilm in 37 °C. The obtained results were validated against monocytes incubated without bacteria. The isolates from patients with chronic kidney disease formed the most adherent biofilm regardless the presence or absence of inflammatory reaction. Adherence of monocytes also increased during therapy with immunosuppressive agents, but monocytes' response was different when cyclosporine or tacrolimus were used. Additionally the presence of inflammatory reaction in patients with kidney disease modified the monocytes response when the immunosuppressive drugs were used. Considering the obtained results, we conclude that the changes of monocytes' morphology in response to biofilm formed by Gram negative rods could become a tool to detect urinary tract infection, especially in those groups of patients, where the knowledge of ongoing inflammation is important and the standard tools fail to detect it.

[Differentiation of spa types and staphylococcal cassette chromosome mec (SCCmec) in clinical methicillin-resistant Staphylococcus aureus isolated in medical sites of Gdansk region]. / Zrónicowanie typów spa i kaset SCCmec u klinicznych szczepów Staphylococcus aureus opornych na metycyline izolowanych w osrodkach regionu gdanskiego.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus bacteria are one of the key etiological factors of hospital-acquired and community-acquired infections. MRSA strains have an ability of causing a broad spectrum infections: from a relatively mild skin infections to severe life-threatening systemic infections. They are characterized by multi-drug resistance, virulence of a number of factors, may clonally spread within the hospitals and between hospitals. METHODS: The study embraced a number of 75 isolates of MRSA isolated from patients of 7 medical sites of the Gdansk region within the period of six months (June to December 2013). Strains have derived from various clinical materials, both of hospitalized patients (n=59) and outpatient (n=16). The isolates were tested for the susceptibility to antimicrobial agents accordance with the guidelines EUCAST. To estimate of the variability of occurrence of S. aureus clones used were standard spa gene, consisting in the amplified polymorphic region of the X gene encoding the protein A gene (spa). After receiving the results, a spa types were identified using international database Ridom Spa Server (www.spaserver.ridom.de). To determine the polymorphism cassette carrying the inecA gene from MRSA strains, used typing five major chromosomal cassette SCCmec (I-V) by multiplex PCR. RESULTS: MRSA population genetic analysis carried out on the basis of typing SCCmec cassettes and spa gene has showed a predominance of strains with SCCmec type II casette (46.7%) and SCCmec IV casette (38.7%). Less frequently detected were strains containing SCCmec I cassette (12.0%) and SCCmec III cassette (2.6%). Spa typing revealed the presence of 13 gene types in MRSA. The most frequently observed spa types were: t151 (24.0%), t003 (16.0%) in strains of the SCCmec II cassette and t437 (16.0%) and t008 (14.8%) in the isolates with SCCmec cassette IV, whereas staphylococcus with the type of spa t011 (12.0%) had SCCmec cassette I. CONCLUSIONS: In our population most frequent strains cassette SCCmec II (46.7%), in most representing types of spa t151 (51.4%) and t003 (34.3%), generally resistant not only to ß-lactam antibiotics, but as erythromycin, clindamycin and norfloxacin (82.8%), the more frequently they were isolated from patients than a hospital outpatient centers. The strains SCCmec IV that represent the majority of outpatient centers (68.8%), the most represented type t437 (41.4%) and often occurred in hospital centers.

The influence of nutritional habits, body mass index and intestinal microbiota in mastocytosis on clinical symptoms using conventional culture and next generation sequencing.

BACKGROUND: Mastocytosis is a rare neoplastic disease of the bone marrow associated with the proliferation and accumulation of mast cells in various internal organs, including the gastrointestinal tract. There are few studies describing the gut microbiome of patients with mastocytosis using next generation sequencing supported using traditional culture methods. The aims of the study were, firstly, the determination of nutrition habits, composition of the intestinal microflora and BMI in mastocytosis, and secondly, analysis of mastocytosis severity and symptoms depending on the composition of the intestinal microflora. METHODS: The study included 47 patients with indolent systemic mastocytosis and 18 healthy controls. All participants gave their informed consent to participate in the study. The study consisted of 3 parts: I-clinical assessment, II - examination of the intestinal microflora using the biochemical method, III - 16S rRNA sequencing. RESULTS: The nutrition habits and BMI of mastocytosis patients were similar to controls; however, most patients with mastocytosis had a low dietary vitamin and mineral content. As many as 94.5% of patients had too little fiber intake and mineral content. The most common cause of the abnormal stool test result with traditional culture was a titer of E. coli <106 . The low richness of microbiota species indicated by the Simpson index was observed in mastocytosis, p = 0.04. There were no significant differences in the composition of the intestinal microflora depending on the type of mastocytosis; however, the tryptase level correlated with the amount of Suterella, Barnesiellaceae, Eubacterium, Odoribacter, and Anaerostipes. CONCLUSIONS: The nutritional habits and BMI of mastocytosis patients are similar to the general population, except for too little fiber intake and mineral content. The gastrointestinal symptoms of mastocytosis patients may be related to the low richness of microbiota species and the amount of Suterella, Barnesiellaceae, Eubacterium, Odoribacter, Anaerostipes, which correlated with tryptase levels.

[Spa types and antibiotic resistance of Staphylococcus aureus bloodstream isolates obtained form patients of the University Clinical Center in Gdansk]. / Typy spa i lekoopornosc szczepów Staphylococcus aureus izolowanych z krwi pacjentów Uniwersyteckiego Centrum Klinicznego w Gdansku.

INTRODUCTION: Staphylococcus aureus is a leading cause of bloodstream infections. For epidemiological investigations of this bacteria spa genotyping is used as the method which has a high discriminatory power and gives results that can be easily compared between laboratories. In contrast to methicillin-resistant S.aureus (MRSA), relatively little is known about spa types among methicillin-susceptible strains (MSSA). We used spa typing and antibiotic resistance patterns analysis for retrospective study of S.aureus bloodstream isolates population from the University Clinical Centre (UCC) in Gdansk. METHODS: The study was performed on 53 isolates from patients of 19 different units/ departments of the UCC. The isolates were tested for the susceptibility to antimicrobial agents. Spa typing was performed on the basis of the sequence analysis of the polymorphic X region of the protein A gene (spa) amplified form the isolates. Spa types were determined by Ridom Staph Type software and were clustered into spa-CCs (clonal complexes) using the algorithm BURP-based upon repeat pattern. MLST (Multilocus Sequence Typing) clonal complexes were predicted from BURP analysis by the Ridom SpaServer database. In MRSA the staphylococcal chromosomal casette (SCC) mec was determined, RESULTS: Spa-typing yielded 26 types. Six spa-CC and seven singletons were identified. The most frequent was spa-CC021involving 38% of isolates. The CC021 consisted of 7 spa types and the most common was t021 corresponding with MLST-CC30. The second frequent was singleton, related to MLST-CC1, with only one type t127. There were 3 MRSA isolates in the population. The MRSA strains were identified as different spa types: t003/ SCCmecII, t008/SCCmecIV and clonally related to MSSA t032/SCCmecIV. No one MRSA strains belonged to spa-CC021. CONCLUSIONS: The spa clonal cluster corresponding with widely distributed among invasive S.aureus strains in Europe MLST-CC30 was found as the most frequent among S.aureus bloodstream isolates from the UCC. Occurrence of spa types which had a genetic background common to well known MRSA clonal lineages was observed.

Changes in the Protein Profile in Staphylococcal Strains from Patients Infected with the SARS-CoV-2 Virus.

Staphylococcus aureus strains are particularly often isolated from patients with SARS-CoV-2 infection. The aim of the current research was to determine whether the SARS-CoV-2 virus infection affects the protein profile of S. aureus. Bacteria were isolated from the forty swabs collected from the patients in the hospitals of the Pomeranian region. MALDI-TOF MS spectra were obtained using a Microflex LT instrument. Twenty-nine peaks were identified. The peak (2,430) is described here for the first time and was unique for the isolates from patients infected with the SARS-CoV-2 virus. These results support the hypothesis of bacterial adaptation to the conditions caused by viral infection.

[Sensitivity for antibiotics among the E. coli strains isolated from patients with different clinical stage of infection]. / Antybiotykowrazliwosc szczepów E. coli wyizolowanych z lozyska krwionosnego pacjentów w róznym stopniu zaawansowania klinicznego zakazenia krwi.

Enteric rods are the microorganisms most commonly isolated from blood of hospitalized patients. Bloodstream infections caused by them are associated with significant patient mortality. The aim of the study was analysis of clinical course and evaluation of clinical response on bloodstream infection caused by Escherichia coli. Microorganisms were evaluated for sensibility for antibacterial drugs. For that reason MIC (Minimal Inhibitory Concentration) of antibiotics from different groups was determined for E. coli strains isolated from patients with different clinical stage of infection. No significant differences were shown in sensitivity for antibiotics and MIC among the E. coli strains in correlation with clinical condition of studied patients.

Microbiological, Clinical and Radiological Aspects of Diabetic Foot Ulcers Infected with Methicillin-Resistant and -Sensitive Staphylococcus aureus.

Diabetic foot ulcer (DFU) is one of the most common chronic complications of diabetes. This study aimed to assess the factors with an impact on the infection of diabetic foot ulcers by methicillin-resistant S. aureus and to evaluate the influence of methicillin resistance on the frequency of osteitis (based on classic X-ray images). A total of 863 patients suffering from DFU were analyzed during the study period. Out of 201 isolated S. aureus cases, 31 (15.4%) were methicillin-resistant (MRSA). MRSA infections were associated with a higher incidence of osteitis compared to MSSA infections (p << 0.0001), both the occurrence of smaller (<50%)) and greater (>50%) inflammatory bone changes (p << 0.0001). Furthermore, MRSA occurred significantly more frequently in men than in women (p < 0.01) and more often among patients with type 2 diabetes than among patients with type 1 diabetes (p < 0.05). MRSA were isolated statistically less often in overweight patients than in patients with normal BMI (p < 0.05). DFUs infected with MRSA were significantly more frequently associated with the presence of Pseudomonas sp. and other non-fermenting bacilli than those infected with MSSA (p < 0.05). To conclude, osteitis incidence is related to MRSA infection in patients with diabetic foot ulcers; thus, patients infected by S. aureus should be closely monitored in the course of using antibiotics and treated with narrow-spectrum antibiotics.

Distribution and antibiotic-resistance of different Staphylococcus species identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) isolated from the oral cavity.

BACKGROUND: The use of antibiotics in dentistry is associated with the emergence and spread of antibiotic-resistant microorganisms, including commensal staphylococci. METHODS: A total of 367 oral samples were collected, from which staphylococci were isolated and identified by using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF). The antibiotic susceptibility of the isolates was determined and molecular characteristics for methicillin-resistant staphylococci was performed. RESULTS: A total of 103 coagulase-negative staphylococci (CoNS), among them S. warneri, S. haemolyticus, S. saprophyticus, S. pasteuri, S. epidermidis, S. hominis, S. xylosus, S. equorum, S. kloosii, S. succinus, S. cohnii, and S. simulans, were confirmed by MALDI-TOF. Resistance to most tested antibiotics was statistically higher in CoNS than in S. aureus isolates (P-value < 0.05). CoNS isolates showed high resistance to penicillin (S. saprophyticus 88.9%), erythromycin (S. haemolyticus 84.6%), fusidic acid (S. saprophyticus 77.8%), co-trimoxazole (S. epidermidis 71.4%), gentamicin (S. warneri 63.8%), and tetracycline (S. saprophyticus 55.6%). Multidrug resistance was largely observed, especially among S. haemolyticus and S. saprophyticus species. Methicillin-resistance in S. haemolyticus (38.5%), S. saprophyticus (22.2%) and S. aureus (13.5%) was associated with the presence of the mecA gene and SCCmec type IV or V. CONCLUSION: Coagulase-negative staphylococci, especially S. haemolyticus and S. saprophyticus, seem to be a reservoir of methicillin resistance and multidrug resistance in the oral cavity.

Genetic Background and Antibiotic Resistance Profiles of K. pneumoniae NDM-1 Strains Isolated from UTI, ABU, and the GI Tract, from One Hospital in Poland, in Relation to Strains Nationally and Worldwide.

In recent years, there has been an observed increase in infections caused by carbapenem-resistant Klebsiella pneumonia (Kp) strains. The aim of this study was the phenotypic and genotypic analysis of eight K. pneumoniae NDM (Kp NDM) isolates, recovered in Poland during the years 2016 and 2018 from seven patients with urinary tract infections (UTIs), asymptomatic bacteriuria (ABU), or colonization of the gut. PCR melting profile genotyping indicated a close relationship between the strains derived from 2018, which were not related to the strain isolated in 2016. WGS results were analyzed in relation to international Kp isolates. Clonal and phylogenetic analyses were performed based on multilocus sequence typing (MLST) and single nucleotide polymorphisms (SNPs) of the core genome. The metallo-ß-lactamase was assigned to the NDM-1 type and the sequence was identified as ST11. Eleven antimicrobial resistance genes were detected, mostly from plasmid contigs. Unprecedented profiles of plasmid replicons were described with the IncFII/pKPX-1 dominant replicon. In terms of the KL24 and O2v1 capsular antigen profiles, these isolates corresponded to Greek strains. Strains isolated from UTI, ABU, and colonization GI tract patients were not carrying environment-specific virulence genes. Based on the assessment of strain relationships at the genome level and their direction of evolution, the international character of the sublines was demonstrated, with a documented epidemic potential in Poland and Greece. In conclusion, some groups of patients, e.g., renal transplant recipients or those with complicated UTIs, who are frequently hospitalized and undergoing antibiotic therapy, should be monitored not only for the risk of UTI, but also for colonization by Kp NDM strains.

Patient Safety Related to Microbiological Contamination of the Environment of a Multi-Profile Clinical Hospital.

Nosocomial infections pose a serious burden for hospitals, patients, and the entire society. The aim of the study was to assess the microbiological cleanliness of the hospital environment through quantitative and qualitative analysis of microbiological contamination of air and surfaces in inpatient treatment facilities, based on the example of a large clinical hospital in Poland. Data were collected between 2012 and 2018 in premises of a large teaching hospital in Gdansk using the sedimentation method and the impact method using the Aerideal apparatus (Biomerieux). In the analyzed clinical center, the microbiological cleanliness tests in most of the hospital rooms in the analyzed period showed an acceptable number of saprophytic microorganisms. Of all the tested samples, 1159 (21.8%) were positive, indicating the presence of microorganisms in the tested sample. Species potentially pathogenic for hospital patients were identified, constituting 20.8% of all positive samples (4.6% of all samples). Significantly higher proportion of microorganisms potentially dangerous to patients were isolated from sanitary facilities. Due to the potentially pathogenic microorganisms detected in the tested samples, the authors suggest that in the analyzed hospital, the areas requiring a specific level of microbiological purity should be designated and described, with [specifically] defined cleaning and disinfection protocols.

Linezolid-resistant Enterococcus faecium strains isolated from one hospital in Poland -commensals or hospital-adapted pathogens?

One of the most pressing problems of enterococci infections is occurring resistance to linezolid, which is an antibiotic used in the treatment of infections caused by vancomycin-resistant strains (VRE). The main objective of our research was to investigate the relationship of 19 linezolid-resistant E. faecium isolates from 18 patients hospitalized at Clinical Hospital in Gdansk (Poland). One of the LZDREF was isolated in 2003 (K2003), and another 18 were collected from 2013 to 2017. Genotyping with PCR MP method indicated 14 main unrelated genetic profiles and no association with K2003 strain. Two isolates with the same genotype and genetically closely related two sub-types (2 isolates for each sub-type) were hospital-derived colonizations of patients. The other unrelated genotypes were discussed in the context of colonization, nosocomial infections, and commensal origin, taking into account prior exposure to linezolid. We determined the presence of a point mutation G2576T in six loci of 23S rDNA. There was also a significant correlation (p<0.0015) between the presence of MIC>32 value and the presence of G2576T point mutation on the sixth rrn. We also detected 5 virulence genes for all isolates: gelE, cylA, asa1, hyl, esp. Correlation (p≤0.0001) was observed between the presence of gelE gene encoding gelatinase and two other genes: cylA and asa1 encoding cytolysin and collagen binding protein responsible for aggregation of bacterial cells, respectively. Significant correlation was also observed between asa1 and cfr genes encoding 23S rRNA rybonuclease responsible for resistance to PhLOPSA antibiotics (p = 0.0004). The multidimensional analysis has also shown the correlation between cfr gene and GI-tract (p = 0, 0491), which suggests horizontal gene transfer inside the gut microbiota and the risk of colonization with linezolid-resistant strains without previously being treated with the antibiotic. The patient could have been colonized with LZDRVREF strains which in the absence of competitive microbiota quickly settle in ecological niches favourable for them and pose a risk for the patient.

Dissemination Of t437-SCCmecIV And Coagulase-Negative t037-SCCmecIII Types Among Borderline Oxacillin-Resistant Staphylococcus aureus Isolated From Skin Infections And Diabetic Foot Ulcers.

BACKGROUND: In a recent decade, the occurrence of S. aureus isolates with low-level oxacillin resistance, have been reported increasingly. The aim of this study was to estimate the prevalence of S. aureus with low-level of oxacillin resistance and to determine their molecular characteristics, including spa types, SCCmec types and presence of toxin genes. METHODS: A total of 249 S. aureus strains were analyzed. Antimicrobial susceptibility was preliminarily tested by the disk diffusion method, and further was verified with the E-test and agar dilution methods. All borderline oxacillin-resistant strains (BORSA) were screened for the mecA gene and virulence factors, including Panton-Valentine leukocidin (PVL). Staphylococcal cassette chromosome mec (SCCmec) typing and spa typing were also carried out. RESULTS: Twelve (4.8%) borderline oxacillin-resistant strains with MIC ≤4 µg/mL were identified. Almost all strains (11/12) were oxacillin-susceptible methicillin resistant S. aureus carrying mecA gene (OS-MRSA). Among the 12 bordeline strains, five spa types (t437, t037, t015, t216, t267) and two SCCmec types (III, IV) were identified, with the most prevalent being t437-SCCmecIV pvl-positive. The second most frequent spa type, t037-SCCmecIII, was sea-positive and did not produce coagulase. The majority of borderline strains originated from skin infections and diabetic foot ulcers and were multidrug-resistant (macrolides, lincosamides and chloramphenicol). CONCLUSION: This study demonstrated that S. aureus with borderline resistance to oxacillin represented primarily SCCmecIV spa type t437 and coagulase-negative SCCmecIII spa type t037 and were isolated from skin infections and diabetic foot ulcers.

Host and pathogen factors in Klebsiella pneumoniae upper urinary tract infections in renal transplant patients.

PURPOSE: To analyse the role of virulence factors (VFs) and host in Klebsiella pneumoniae upper urinary tract infections (UTIs) in renal transplant (RTx) recipients. METHODOLOGY: Clinical and demographic data were registered prospectively. Phylogenetic background of K. pneumoniae isolates was analysed by PCR melting profiles (MP) and the following VFs genes: fimH-1, uge, kpn, ycfM, mrkD, rmpA, magA, hlyA, cnf-1, irp-1, irp-2, fyuA, entB, iutA, iroN by PCR. RESULTS: We studied urine cultures and clinical data from 61 episodes of K. pneumoniae UTI in 54 RTx recipients. There were 32 cases of AB (53%), 10 cases of lower UTI (16%), 19 cases of AGPN (31%), including six cases of bacteraemia. In total, 74 % of strains were extended-spectrum beta-lactamase+, and there were two carbapenemase-producing strains. PCR MP typing showed a diverse population with 52 different genetic profiles of K. pneumoniae. Analysis of the DNA profiles indicated 45 unrelated, unique genotypes and 7 related (16 isolates from 15 patients) genotypes. Urine flow impairment emerged as an independent predictor of K. pneumoniae upper UTIs (OR 14.28, CI 2.7-75.56, P 0.002), while we did not find any association between the profile of VFs and developing upper UTIs. The prevalence of the uge gene was lower in RTx patients on everolimus when compared to isolates from patients not receiving mTOR inhibitors (33.3 % vs 82.8 % P<0.05). CONCLUSIONS: K. pneumoniae upper UTI may be a marker of urine flow impairment. Bacterial VFs could not discriminate between upper and lower UTIs. However, immunosuppression may influence the selection of particular VFs.

Current epidemiology and practice patterns in prevention and treatment of PD-related infections in Poland.

BACKGROUND: Peritoneal dialysis (PD) related infections are associated with technique failure and mortality. The aim of this multicentre study was to examine epidemiology, treatment and outcomes of PD-related infections in Poland as well as practice patterns for prevention of these complications in the context of current ISPD recommendations. METHODS: A survey on PD practices in relation to infectious complications was conducted in 11 large Polish PD centres. Epidemiology of peritonitis and exit-site infections (ESI) was examined in all patients treated in these units over a 2 year period. RESULTS: The study included data on 559 PD patients with 62.4% on CAPD. Practice patterns for prevention of infectious complications are presented. The rate of peritonitis was 0.29 episodes per year at risk, with Gram positive microorganisms responsible for more than 50% of infections and 85.8% effectively treated. Diagnosis and treatment followed ISPD guidelines however most units did not provide an anti-fungal prophylaxis. Although neither of the centres reported routine topical mupirocin on catheter exit-site, the rate of ESI was low (0.1 episodes per year at risk), with Staphylococcus aureus as most common pathogen and full recovery in 78.3% of cases. CONCLUSION: The study shows rewarding outcomes in prevention and treatment of PD-associated infections, mainly due to a thorough compliance with the current ISPD guidelines, although some deviations from the recommendations in terms of practice patterns have been observed. More studies are needed in large numbers of patients to differentiate the importance of specific recommendations and further support the guidelines.

PCR melting profile method for genotyping analysis of vancomycin-resistant Enterococcus faecium isolates from Hematological Unit patients.

A number of Enterococcus strains with high-level inducible resistance to vancomycin have been identified, and the relative incidence of these strains has increased significantly in the last years. The first outbreak caused by vancomycin-resistant enterococci in Poland was reported in 1999. Vancomycin-resistant Enterococcus faecium is known for its propensity to cause infections which are difficult to eradicate. In this study, we determined the genetic similarities between vancomycin-resistant E. faecium isolates consecutively recovered from single patients to assess the duration of infection or colonization. The isolates taken in the study were identified by the conventional methods as E. faecium. PCR melting profile (PCR-MP) and pulsed-field gel electrophoresis (PFGE) typing revealed that the isolates belonged to six distinct genotypes and that two of them were predominant. Consecutive E. faecium isolates with identical genotypes were found in 7 of 12 (58.0%) patients. The delay between the times of recovery of the first and last isolates of identical genotypes from each patient was from 9 days to about 1 year. In six patients, paired blood and non-blood isolates showed identical genotypes. Data presented here demonstrate the complexity of the epidemiological situation concerning vancomycin-resistant enterococci that may occur in a single medical ward. We also show for the first time the evaluation of PCR-MP technique in enterococci strains differentiation and we revealed that there is at least a similar power of discrimination between the present gold-standard REA-PFGE and a PCR-MP method.

[Nosocomial bacteremia]. / Bakteriemie szpitalne.

Nosocomial bacteremia according to different authors can be divided in to bacteremia persistent, polymicrobial bacteremia, alternate bacteremia, breakthrough bacteremia, different bacteremia. Developing techniques of microbiological and molecular diagnostics induce us to verify the definition and partition of term "bacteremia".

[Escherichia coli a forgotten pathogen in septicemia]. / Escherichia coli zapomniany patogen posocznic.

The aim of the retrospect study was to analyse the incidence of E. coli bacteremia in eight wards of SPSK 1 ACK AM in Gdansk from 2002 to 2004. We analyzed the incidence of bacteremia, patients outcome, source of infection and antimicrobial susceptibility. During the study period we detected 268 patients with E. coli bacteremia (8,0% of all bacteremic patients). 11,2% of them died within 24-48h after positive blood culture. Incidence of bacteremia was 1,7/1000 patients and the highest level achieved in Hematology Unit--33,2. The main portal of entry was genitourinary tract (24,3%) and gastrointestinal tract (21,8%). The strains (n=263) were least susceptible to ampicillin (33,3%), co-trimoxazole (68,4%), amoxycillin with clavulanic acid (69,3%) and ciprofloxacine (78,9%).

[Microorganisms analysis in Public Clinical Hospital in Gdansk over three years 2001-2003]. / Analiza wyników diagnostycznych badan mikrobiologicznych u pacjentów Szpitala Klinicznego w Gdansku, w latach 2001-2003.

The aim of the study was to analyse the changes in occurence of microorganisms and antibiotic usage in tertiary care hospital over 3 years. We analysed the results of microbiological records from laboratory inforation systems from 2001 to 2003. Over the study period there was about 40% increase of specimens received in the laboratory mainly due to another hospital incorporation. The relations between different groups of microorganisms was stable, Gram negatives 44,4%-46,3%, Gram positives 37,3%-40,3%, yeasts 7,0%-8,1%. There was a decrease in MRSA from 0,6% to 0,2% and carbapenem resistant Pseudomonas aeruginosa (CRPA) isolations from 2,0% to 0,7%, however the reverse was true for VRE, increase from 0,3% to 2%. ESBL-producing bacteria were isolated from about 4% of Enterobacteriaceae throughout the study. The analysis of blood cultures revealed over 60% deacrease in P. aeruginosa bacteremia and stable incidence of Escherichia coli (7%) and Staphylococcus aureus (6,5%) bacteremia. Increased usage of cephalosporins and fluoroquinolones was accompanied by the decrease in carbapenems and penicillins. In most cases there were no significant changes in occurence of main groups of microorganisms. Some multidrug resistant bacteria like MRSA and CRPA are no longer a problem in our hospital. Others like VRE, ESBL and Acinetobacter still cause concern due to high colonisation or infection rate. The usage of some antibiotic groups increased, another decreased and finally some like aminoglicosides and glicopeptides remained stable.

Evaluation of a novel method based on amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) for typing strains of vancomycin-resistant Enterococcus faecium.

In the search for an effective DNA-typing technique for use in hospital epidemiology, the performance and convenience of a novel assay based on the fingerprinting of bacterial genomes by amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) was tested. A large number of vancomycin-resistant Enterococcus faecium (VREM) isolates from haematological ward patients of the Clinical Hospital in Gdansk were examined. We found that ADSRRS fingerprinting analysis is a rapid method that offers good discriminatory power. The method demonstrated also excellent reproducibility. The usefulness of the ADSRRS fingerprinting method for molecular typing was compared with pulsed field gel electrophoresis (PFGE) method, which is currently considered the gold standard for molecular typing of isolates recovered from patients and the environment in the course of investigation and control of nosocomial outbreaks. Clustering of ADSRRS fingerprinting data matched pulsed field gel electrophoresis data. The features of ADSRRS fingerprinting technique is discussed in comparison with conventional methods. Data presented here demonstrate the complexity of the epidemiological situation concerning VREM that may occur in a single medical ward.

Emerging linezolid-resistant, vancomycin resistant Enterococcus faecium from a patient of a haematological unit in Poland.

The oxazolidinone antimicrobial, linezolid, has been approved for the treatment of infections caused by various gram-positive bacteria, including vancomycin-resistant enterococci (VRE). This is the first report on isolation of a linezolid resistant, vancomycin-resistant Enterococcus faecium (LRVREF) strain in Poland, from a Haematological Unit patient in the Clinical Hospital in Gdansk. PCR-RFLP analysis of rDNA and allele-specific PCR of the domain V region of the 23S ribosomal RNA gene demonstrated the presence of the G2576U mutation previously reported to be associated with linezolid resistance. Both assays detected heterozygous in this position.