Antimicrobial susceptibility and prevalence of extended-spectrum beta-lactamases in clinical strains of Klebsiella pneumoniae isolated from pediatric and adult patients of two Polish hospitals.

Klebsiella pneumoniae due to the presence of multiple antibiotic resistance mechanisms is one of the most threatening human pathogens nowadays. The aim of the study was to characterize antimicrobial susceptibility, presence of resistance mechanisms and the prevalence of selected genes encoding ESBLs in 170 K. pneumoniae isolates recovered from children and adults hospitalized in two Polish medical centers from 2008 to 2015. The phenotypic identification of strains was confirmed by amplification of mdh gene. ESBLs, metallo-beta- lactamases, Klebsiella pneumoniae carbapenemases and OXA-48 were detected using phenotypic tests. The blaCTX-M-1, blaTEM and blaSHV ESBL genes were amplified by PCR. Pediatric K. pneumoniae isolates displayed significantly higher resistance to piperacillin/tazobactam, cefoxitin, imipenem, amikacin and ciprofloxacin than strains obtained from adults (P<0.05). The presence of ESBLs, OXA-48, KPC and MBL was confirmed in 80.6%, 21.8%, 8.2% and 2.4%, respectively, of the tested strains. The CTX-M-1 enzymes were predominant (91.2%), followed by TEM (63.5%) and SHV (11.8%). The blaTEM was significantly more common in adults than in children (P<0.05). Dual or triple bla genes were observed in 55.9% and 8.2% of K. pneumoniae isolates. Further local epidemiological studies are required to monitor the dissemination of multidrug-resistant K. pneumoniae strains.

Prevalence of human pathogens of the clade Nakaseomyces in a culture collection-the first report on Candida bracarensis in Poland.

Human pathogens belonging to the Nakaseomyces clade include Candida glabrata sensu stricto, Candida nivariensis and Candida bracarensis. Their highly similar phenotypic characteristics often lead to misidentification by conventional laboratory methods. Therefore, limited information on the true epidemiology of the Candida glabrata species complex is available. Due to life-threatening infections caused by these species, it is crucial to supplement this knowledge. The aim of the study was to estimate the prevalence of C. bracarensis and C. nivariensis in a culture collection of C. glabrata complex isolates. The study covered 353 isolates identified by biochemical methods as C. glabrata, collected from paediatric and adult patients hospitalised at four medical centres in Southern Poland. The multiplex PCR was used to identify the strains. Further species confirmation was performed via sequencing and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) analysis. One isolate was recognised as C. bracarensis (0.28%). To our knowledge, it is the first isolate in Poland. C. glabrata sensu stricto species has been confirmed for all the remaining isolates. No C. nivariensis was found. Our study has shown that the prevalence of C. nivariensis and C. bracarensis strains is infrequent. However, it should be emphasised that the incidence of these strains may differ locally and depend on environmental factors and the population.

Nested PCR for the detection of Aspergillus species in maxillary sinus samples of patients with chronic sinusitis.

BACKGROUND: Fungal rhinosinusitis has become an increasingly recognized disease, being Aspergillus species responsible for most of the cases. Its diagnosis is quite difficult because of the non-specific symptoms and low sensitivity of the current diagnostic methods. AIMS: An Aspergillus-specific nested polymerase chain reaction (PCR) assay using biopsy specimens taken from the maxillary sinuses was performed in order to assess its usefulness. Conventional diagnostic methods (histology and culture) were also carried out. METHODS: A case-control study was performed in the Institute of Stomatology, Jagiellonian University in Kraków, between 2011 and 2014. The case group consisted of 21 patients with suspected rhinosinusal mycetoma while the control group included 46 patients with no suspicion of fungal rhinosinusitis. The two-step PCR assay amplified an Aspergillus specific portion of the 18S rRNA gene. Interval estimation of sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were calculated to assess the diagnostic test performance. The agreement between the PCR and the other tests was evaluated using the Kappa coefficient (k). RESULTS: Ninety percent of the samples obtained from patients diagnosed with mycetoma yielded positive PCR results. The PCR showed almost perfect concordance with histology (k=0.88). Sensitivity, specificity, PPV and NPV estimates were 90%; 95% CI: (55.5-99.7%), 98.3%; 95% CI: (90.9-100%), 90%; 95% CI: (55.5-99.7%) and 98.3%; 95% CI: (90.9-100%), respectively. One clinical sample showed growth of Aspergillus fumigatus and positive PCR despite the negative histological examination. CONCLUSIONS: Nested PCR assay is a promising diagnostic tool to evaluate the presence of Aspergillus in the tissue of maxillary sinus from patients with suspicion of sinus aspergillosis.

Nested PCR for the detection of Aspergillus species in maxillary sinus samples of patients with chronic sinusitis / PCR con cebadores internos para la detección de especies de Aspergillus en muestras de seno maxilar de pacientes con sinusitis crónica

Background: Fungal rhinosinusitis has become an increasingly recognized disease, being Aspergillus species responsible for most of the cases. Its diagnosis is quite difficult because of the non-specific symptoms and low sensitivity of the current diagnostic methods. Aims: An Aspergillus-specific nested polymerase chain reaction (PCR) assay using biopsy specimens taken from the maxillary sinuses was performed in order to assess its usefulness. Conventional diagnostic methods (histology and culture) were also carried out. Methods: A case-control study was performed in the Institute of Stomatology, Jagiellonian University in Kraków, between 2011 and 2014. The case group consisted of 21 patients with suspected rhinosinusal mycetoma while the control group included 46 patients with no suspicion of fungal rhinosinusitis. The two-step PCR assay amplified an Aspergillus specific portion of the 18S rRNA gene. Interval estimation of sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were calculated to assess the diagnostic test performance. The agreement between the PCR and the other tests was evaluated using the Kappa coefficient (k). Results: Ninety percent of the samples obtained from patients diagnosed with mycetoma yielded positive PCR results. The PCR showed almost perfect concordance with histology (k=0.88). Sensitivity, specificity, PPV and NPV estimates were 90%; 95% CI: (55.5-99.7%), 98.3%; 95% CI: (90.9-100%), 90%; 95% CI: (55.5-99.7%) and 98.3%; 95% CI: (90.9-100%), respectively. One clinical sample showed growth of Aspergillus fumigatus and positive PCR despite the negative histological examination. Conclusions: Nested PCR assay is a promising diagnostic tool to evaluate the presence of Aspergillus in the tissue of maxillary sinus from patients with suspicion of sinus aspergillosis

Antecedentes: La rinosinusitis fúngica se ha convertido en una enfermedad cada vez más frecuente y el género Aspergillus es el causante de la mayoría de los casos. Su diagnóstico es relativamente difícil debido a la inespecificidad de los síntomas y a la baja sensibilidad de los métodos de diagnóstico actuales. Objetivos: Evaluar la eficacia de un ensayo de reacción en cadena de la polimerasa (RCP), con cebadores internos y específica de Aspergillus en muestras de biopsia tomadas de los senos maxilares de algunos pacientes, y compararla con la eficacia de los métodos de diagnóstico convencionales (histología y cultivo). Métodos: Se realizó un estudio de casos y controles en el Instituto de Estomatología de la Universidad Jaguelónica de Cracovia entre 2011 y 2014. El grupo de casos estaba formado por 21 pacientes en que se sospechaba rinosinusitis por micetoma mientras que el grupo control estaba compuesto por 46 pacientes sin sospecha de rinosinusitis fúngica. El ensayo de PCR en dos etapas amplificó una porción específica del gen 18S rRNA de Aspergillus. Se obtuvieron estimaciones de la sensibilidad, la especificidad y de los valores predictivos positivo (VPP) y negativo (VPN) para evaluar el rendimiento de la prueba. La concordancia entre la PCR y las otras pruebas realizadas se evaluó utilizando el coeficiente kappa (k). Resultados: El 90% de las muestras obtenidas de pacientes diagnosticados de micetoma mostró resultados positivos en la PCR, con una concordancia casi perfecta de este método con la histología (k=0,88). Las estimaciones de sensibilidad, especificidad, VPP y VPN fueron las siguientes: 90%, IC95% (55,5-99,7%); 98,3%, IC95% (90,9-100%); 90%, IC95% (55,5-99,7%) y 98,3%, IC95%: (90,9-100%), respectivamente. Aspergillus fumigatus se aisló en el cultivo de una muestra clínica, además de obtenerse un resultado positivo por PCR de dicha muestra a pesar de que el examen histológico fue negativo. Conclusiones: El ensayo de PCR con cebadores internos es una herramienta de diagnóstico prometedora para evaluar la existencia de Aspergillus en tejidos del seno maxilar de pacientes en que se sospeche aspergilosis sinusal

Preliminary antifungal activity assay of selected chlorine-containing derivatives of xanthone and phenoxyethyl amines.

The aim of this study was to evaluate antifungal activity in a diverse group of chlorine-containing xanthone and phenoxyethyl amine derivatives - and to select the most promising compounds for further studies. The antifungal efficacy of 16 compounds was tested with qualitative and quantitative methods against both reference and clinical strains of dermatophytes, moulds and yeasts. The disc-diffusion method has demonstrated that from 16 tested compounds, 7 possess good antifungal activity against dermatophytes and/or moulds while none of them has shown good efficacy against yeasts or bacterial strains. The most active compounds (2, 4, 10, 11, 12, 15, 16) were tested quantitatively by broth dilution method to obtain MIC values. The MIC values against dermatophytes ranged from 8 to 64 µg/ml. Compound 2 was the most active one against dermatophytes (MIC 50 and MIC 90 were 8 µg/ml). The MIC values for moulds ranged from 16 to 256 µg/ml. Compound 4 was the most active one against moulds, with MIC 50 and MIC 90 values amounting to 32 µg/ml. Among the tested compounds, compound 4 (derivative of xanthone) was the most active one and expressed good antifungal efficacy against clinical strains of dermatophytes and moulds. However, another xanthone derivative (compound 2) was the most active and selective against dermatophytes.

Anti-Helicobacter pylori activities of selected N-substituted cinnamamide derivatives evaluated on reference and clinical bacterial strains.

In this study, thirty-five N-substituted derivatives of cinnamic acid amide (cinnamamide) were evaluated for anti-Helicobacter pylori activity using an agar disc-diffusion method. Qualitative screening was performed on a reference H. pylori strain (ATCC 43504), resulting in the identification of the three most active compounds, 8 (R,S-(2E)-3-(4-chlorophenyl)-N-(2-hydroxypropyl)prop-2-enamide, minimal inhibitory concentration, MIC = 7.5 µg/mL), 23 ((2E)-3-(4-chlorophenyl)-N-(2-hydroxycyclohexyl)prop-2-enamide, MIC = 10 µg/mL), and 28 ((2E)-3-(4-chlorophenyl)-N-(4-oxocyclohexyl)prop-2-enamide, MIC = 10 µg/mL). These compounds were further tested on twelve well-characterized clinical strains, yielding MIC values that ranged from 10 to 1000 µg/mL. Preliminary safety assessments of the compounds were made using the MTT viability test for cytotoxicity and Ames test for mutagenicity, which showed them to be generally safe, although compounds 8 and 28 showed mutagenic activity at some of the tested concentrations. The results of this study showed the anti-H. pylori potential of cinnamamide derivatives.

Molecular characterization of Candida isolates from intensive care unit patients, Krakow, Poland / Caracterización molecular de aislamientos de Candida de pacientes en cuidados intensivos en Cracovia, Polonia

Background. Over the last decades, Candida species have emerged as important pathogens in immunocompromised patients. Nosocomial infections are mainly of endogenous origin. Nevertheless, some cases of exogenous candidiasis have also been reported. Aims. The aim of this study was to evaluate the genetic relatedness between Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei and Candida kefyr isolates recovered from intensive care unit (ICU) patients. Methods. A total of 132 Candida clinical isolates (62 C. albicans, 40 C. glabrata, 13 C. tropicalis, 11 C. krusei, 6 C. kefyr), obtained from specimens of endotracheal aspirate, urine and blood taken from patients of a tertiary hospital in Poland, were included in the study. Species identification was performed by PCR method and genetic relatedness was assessed by randomly amplified polymorphic DNA assay (RAPD) with five primers. Results. The RAPD analysis revealed high genetic diversity among the studied Candida isolates, indicating that most of the strains were from endogenous sources. Only two clonal strains of C. glabrata isolated from different patients were observed, suggesting a possible cross-transmission of these pathogens. Conclusions. Our study confirmed the high discriminatory power of the RAPD assay. This genotyping method can be applied to local epidemiological studies of Candida species (AU)

Antecedentes. En las últimas décadas, el hongo Candida se ha convertido en un patógeno importante para los pacientes con trastornos del sistema inmune. Las infecciones nosocomiales son fundamentalmente de origen endógeno; sin embargo, también se han documentado algunos casos de candidiasis exógena. Objetivos. El objetivo del estudio fue evaluar la relación genética entre las cepas de Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei y Candida kefyr aisladas de pacientes en cuidados intensivos. Métodos. Se estudiaron 132 aislamientos de Candida (62 C. albicans, 40 C. glabrata, 13 C. tropicalis, 11 C. krusei, 6 C. kefyr) obtenidos de muestras procedentes de aspirado endotraqueal, orina y sangre tomadas de pacientes de un hospital en Polonia. La identificación de las especies se realizó mediante PCR, y el estudio de la relación genética con el método de amplificación aleatoria de ADN polimórfico (RAPD) con cinco oligonucleótidos. Resultados. El análisis de la amplificación por RAPD mostró una alta diversidad genética entre los aislamientos objeto de estudio, lo que indica que la mayoría de ellos tenían un origen endógeno. Solo se observaron dos cepas clonales de C. glabrata procedentes de diferentes pacientes, lo que evidencia una posible transmisión cruzada de estos patógenos. Conclusiones. Nuestro estudio confirma el alto poder discriminatorio de la técnica RAPD, lo que validaría este método de genotipificación para el estudio de la epidemiología local de especies de Candida (AU)

Molecular characterization of Candida isolates from intensive care unit patients, Krakow, Poland.

BACKGROUND: Over the last decades, Candida species have emerged as important pathogens in immunocompromised patients. Nosocomial infections are mainly of endogenous origin. Nevertheless, some cases of exogenous candidiasis have also been reported. AIMS: The aim of this study was to evaluate the genetic relatedness between Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei and Candida kefyr isolates recovered from intensive care unit (ICU) patients. METHODS: A total of 132 Candida clinical isolates (62 C. albicans, 40 C. glabrata, 13 C. tropicalis, 11 C. krusei, 6 C. kefyr), obtained from specimens of endotracheal aspirate, urine and blood taken from patients of a tertiary hospital in Poland, were included in the study. Species identification was performed by PCR method and genetic relatedness was assessed by randomly amplified polymorphic DNA assay (RAPD) with five primers. RESULTS: The RAPD analysis revealed high genetic diversity among the studied Candida isolates, indicating that most of the strains were from endogenous sources. Only two clonal strains of C. glabrata isolated from different patients were observed, suggesting a possible cross-transmission of these pathogens. CONCLUSIONS: Our study confirmed the high discriminatory power of the RAPD assay. This genotyping method can be applied to local epidemiological studies of Candida species.

Anti-Helicobacter pylori activity of some newly synthesized derivatives of xanthone.

A series of 20 xanthone derivatives was synthesized and evaluated for anti-Helicobacter pylori (H. pylori) activity. Qualitative and quantitative in vitro tests using the Kirby-Bauer method (agar disc-diffusion method) were performed. The tested compounds were screened against clarithromycin- and/or metronidazole-resistant strains of H. pylori. As a reference, Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial strains were examined. On the basis of microbiological assays, xanthones can be considered as potential anti-H. pylori agents. They displayed significant activity against the examined strains, which was higher against the bacteria resistant to metronidazole than clarithromycin. The lowest MIC values ranging up to 20 mg l-1 were observed for the following compounds: 3, 4, 8, 9, 12, 19 (against the metronidazole-resistant strains) and the compound 10 (against the clarithromycin-resistant strain). These preliminary results for screening of xanthone derivatives form a part of an ongoing study of the structure-activity relationships of a large group of compounds. Microbiological assays will be conducted afterwards to determine the mechanism of xanthones' action against H. pylori.

ACTIVITY OF THYME OIL (OLEUM THYMI) AGAINST MULTIDRUG-RESISTANT ACINETOBACTER BAUMANNII AND PSEUDOMONAS AERUGINOSA.

Almost as soon as antibiotics were introduced to treat infectious diseases, it could be observed that bacteria were able to develop resistance against them. Currently, multidrug-resistant strains are being isolated mainly in the hospital environment. These are primarily non-fermenting Gram-negative bacilli, which exhibit both natural and acquired resistance to multiple antibiotics and disinfectants rendering them difficult to eradicate. The development of new, effective and safe substances that prevent troublesome infections is greatly needed to provide alternative therapeutic options for patients. There is increasing interest in drugs of natural origin, including essential oils. It is of particular interest that, although active against many bacterial strains, they do not contribute to antibacterial resistance against their components. The aim of our study was to evaluate the in vino antibacterial activity of thyme oil against multidrug-resistant strains of A. baumannii and P. aeriginosa using the disc diffusion and macrodilution methods. The strains were isolated from patients hospitalized in the years 2013-2014. The in vitto antibacterial activity of thyme oil was assessed by the disc diffusion method and the inhibition zones for the oil at different concentrations, produced against A. baumannii, ranged from 7 to 44 mm. Low level of activity of thyme oil was observed against P. aeruginosa strains. The results of serial dilution tests confirmed the high activity of thyme oil against A. baumannii isolates, expressed as MIC values ranging from 0.25 to 2 µL/mL. These results suggest the need for further studies of antibacterial activity of essential oils, especially against multidrug-resistant bacterial isolates.

Levofloxacin resistance of Helicobacter pylori strains isolated from patients in southern Poland, between 2006-2012.

An increasing resistance of Helicobacter pylori (H. pylori) to antimicrobial agents leads to the need of regional monitoring of the prevalence resistant strains (according to the Maastricht/Florence consensus report, 2012). The aim of the study was to assess the resistance to levofloxacin of H. pylori strains isolated from adult patients of Malopolska region in Poland. Bioptates taken from gastric mucosa during gastroscopy constituted the material for the study. Two hundred ten H. pylori strains were isolated from 811 patients. A majority of strains (171) came from patients before the treatment of H. pylori infections while the remaining 39 strains were isolated from patients after the failed therapy. Susceptibility of H. pylori to levofloxacin was determined by strips impregnated with antibiotic gradient (E-test, bioMerieux). The obtained minimum inhibitory concentration (MIC) values ranged from 0.002 mg/L to 32 mg/L. The percentage of strains resistant to levofloxacin amounted to 8.10% (17/210). Among the group of strains isolated from patients before the treatment, 5.85% (10/171) of H. pylori strains were resistant to levofloxacin. In the group of strains isolated from patients after the treatment 17.95% (7/39) of strains were resistant. The difference in the frequency of H. pylori strains resistant to levofloxacin in patients before and after the treatment of the infection due to H. pylori was statistically significant (p = 0.0297). The low percentage of H. pylori strains resistant to levofloxacin justify that the introduction of a triple therapy with levofloxacin is a good alternative in the treatment of H. pylori infections, especially in regions with high prevalence of H. pylori strains resistant to clarithromycin (> 20%).

Molecular epidemiology of Candida albicans and Candida glabrata strains isolated from intensive care unit patients in Poland.

Over the last decades, Candida spp have been responsible for an increasing number of infections, especially in patients requiring intensive care. Knowledge of local epidemiology and analysis of the spread of these pathogens is important in understanding and controlling their transmission. The aim of this study was to evaluate the genetic diversity of 31 Candida albicans and 17 Candida glabrata isolates recovered from intensive care unit patients from the tertiary hospital in Krakow between 2011-2012. The strains were typed by random amplified polymorphic DNA (RAPD) polymerase chain reaction using five primers (CD16AS, HP1247, ERIC-2, OPE-3 and OPE-18). The results of the present investigation revealed a high degree of genetic diversity among the isolates. No clonal relationship was found among the C. albicans strains, whereas two C. glabrata isolates were identical. The source of Candida infection appeared to be mostly endogenous; however, the presence of two clonal C. glabrata strains suggested the possibility of cross-transmission of these pathogens. Our study confirmed the high discriminatory power of the RAPD technique in the molecular typing of Candida clinical isolates. This method may be applied to the evaluation of transmission routes of pathogenic fungi on a local level.

Molecular epidemiology of Candida albicans and Candida glabrata strains isolated from intensive care unit patients in Poland

Over the last decades, Candida spp have been responsible for an increasing number of infections, especially in patients requiring intensive care. Knowledge of local epidemiology and analysis of the spread of these pathogens is important in understanding and controlling their transmission. The aim of this study was to evaluate the genetic diversity of 31 Candida albicans and 17 Candida glabrata isolates recovered from intensive care unit patients from the tertiary hospital in Krakow between 2011-2012. The strains were typed by random amplified polymorphic DNA (RAPD) polymerase chain reaction using five primers (CD16AS, HP1247, ERIC-2, OPE-3 and OPE-18). The results of the present investigation revealed a high degree of genetic diversity among the isolates. No clonal relationship was found among the C. albicans strains, whereas two C. glabrata isolates were identical. The source of Candida infection appeared to be mostly endogenous; however, the presence of two clonal C. glabrata strains suggested the possibility of cross-transmission of these pathogens. Our study confirmed the high discriminatory power of the RAPD technique in the molecular typing of Candida clinical isolates. This method may be applied to the evaluation of transmission routes of pathogenic fungi on a local level.

PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains.

BACKGROUND: The occurrence of clarithromycin resistance among Helicobacter pylori strains is a major cause of the treatment failure. Resistance to this drug is conferred by point mutations in 23S rRNA gene and the most prevalent mutations are A2143G and A2142G. The aim of the study was to evaluate the occurrence of A2143G and A2142G mutations in a group of H. pylori strains resistant to clarithromycin. MATERIALS AND METHODS: The study included 21 clarithromycin-resistant H. pylori strains collected between 2006 and 2009 in southern Poland. Resistance to clarithromycin was quantitatively tested with the E-test to determine the minimal inhibitory concentration (MIC value). The point mutations of H. pylori isolates were detected by PCR followed by RFLP analysis. RESULTS: The MIC values for clarithromycin for the analyzed strains ranged from 1.5 mg/L to 64 mg/L. Nine H. pylori strains exhibited A2143G mutation and A2142G mutation was found in 9 isolates as well. The results of RFLP analysis of 3 clarithromycin-resistant strains were negative for both mutations. The average MIC values for A2143G and A2142G mutants were 6 and 30 mg/L, respectively. CONCLUSIONS: Frequencies of A2143G and A2142G mutations were the same in all isolates tested. Strains with A2143G mutation exhibited lower MIC values than A2142G mutants. Application of PCR-RFLP method for detection of clarithromycin resistance allows for better and more efficient management of H. pylori infections.

Co-occurrence of carbapenem and aminoglycoside resistance genes among multidrug-resistant clinical isolates of Acinetobacter baumannii from Cracow, Poland.

BACKGROUND: Acinetobacter baumannii is a significant hospital pathogen, possessing a considerable degree of antimicrobial resistance. A. baumannii resistance to carbapenems and aminoglycosides is mostly conferred by class D OXA carbapenemases and aminoglycoside-modifying enzymes, respectively. The aim of this study was to determine the prevalence of selected genes encoding OXA carbapenemases and aminoglycoside-modifying enzymes in multidrug-resistant strains of A. baumannii. MATERIAL AND METHODS: The study included 61 carbapenem-resistant and aminoglycoside-nonsusceptible A. baumannii isolates, collected between 2009 and 2011 in Cracow, Poland. Selected resistance genes, including: blaOXA-51-like, blaOXA-23-like, blaOXA-40-like, blaOXA-58-like, aac(6')-Ih, aac(3)-Ia, aac(3)-IIa, aac(6')-Ib, aph(3')-Ia and aph(3')-VI, were detected by PCR method. RESULTS: The blaOXA-51-like genes were detected in all isolates, while acquired carbapenemase encoding genes were found in 96.7% of tested strains. Presence of blaOXA-40-like and blaOXA-23-like genes was observed among 65.6% and 27.9% of isolates, respectively. Assayed aminoglycoside resistance genes were found to harbor 98.4% of isolates. Among tested strains, we observed the following percentages of resistance determinants: aac(3)-Ia - 78.7%, aph(3')-VI - 78.7% and aph(3')-Ia - 27.9%. Analysis of co-occurrence of carbapenem and aminoglycoside resistance genes revealed the highest percentage of strains possessing blaOXA-40-like, aac(3)-Ia, and aph(3')-VI genes (44.3%). CONCLUSIONS: The blaOXA-40-like and aac(3)-Ia/aph(3')-VI were the most prevalent genes encoding acquired OXA carbapenemases and aminoglycoside-modifying enzymes, respectively, among A. baumannii strains in Cracow, Poland. Genes conferring resistance to carbapenems and aminoglycosides coexisted in the clinical strains of A. baumannii. The phenomenon of A. baumannii resistance indicates the necessity of monitoring for the presence of the resistance genes.

Dermatophytes isolated from superficial fungal infections in Krakow, Poland, between 1995 and 2010.

Superficial fungal infections due to dermatophytes are common over the world and their frequency is constantly increasing. The aim of our study was to discuss fungal infections with frequency of occurrence, clinical stages and aetiology in patients admitted to dermatological ward and microbiological laboratory of the specialist hospital in Krakow. Investigations performed between 1995 and 2010 included the group of 5333 individuals. Dermatophyte infections, confirmed by culture, were revealed in 1007 subjects (18.9%), i.e. in 553 males and 454 females. The most frequent clinical forms of infections were tinea unguium and tinea pedis, caused mainly by Trichophyton rubrum and by Trichophyton mentagrophytes. Tinea corporis, tinea manuum, tinea capitis and tinea cruris constituted a small percentage of infections and the main aetiological factors of these dermatomycoses were also T. rubrum and T. mentagrophytes. Between 1995 and 2000 there were stated small differences in the number of isolated strains of dermatophytes in comparison with the number of examined patients. Since 2006 there has been observed a decrease in number of patients in our hospital with suspected fungal infections, but per cent of positive cultures has remained unchanged in comparison with earlier period.

[Occurrence of alert pathogens in hospital environment. Part I. ESBL-producing enterobacteriaceae strains]. / Wystepowanie patogenów alarmowych w srodowisku szpitalnym. Czesc I. Paleczki z rodziny Enterobacteriaceae wytwarzajace beta-laktamazy ESBL.

INTRODUCTION: Gram-negative bacteria belonging to the family Enterobacteriaceae cause severe and difficult to treat nosocomial infections. Strains of different species that produce extended-spectrum beta-lactamases (ESBL) were classified as alert pathogens. The purpose of this study was to analyze the occurrence and determination of antimicrobial susceptibility patterns of 134 ESBL-producing Enterobacteriaceae strains. METHODS: 96 (72%) isolates of Klebsiella pneumoniae and 38 (28%) isolates of Escherichia coli, were cultured from patients of Specialistic Hospital in Krakow, in the period from 2008 to 2010. Bacterial identification and antimicrobial susceptibility testing were performed by automated system Vitek 2 Compact (bioMerieux, Poland). Condition for inclusion in the study was the production by the strains of beta-lactamases with extended substrate spectrum (ESBL), which was confirmed using the automated method (Vitek 2 Compact system) and the disc-diffusion assay (DDST). Taken into consideration the first isolate from the patient. RESULTS: Bacilli of the species K. pneumoniae ESBL(+) were mainly isolated from respiratory tract samples (46%), urine (27%) and blood (12%). The dominant divisions in terms of frequency of isolation of these pathogens were anesthesiology and intensive care (42%), neurology and brain strokes (16%) and internal medicine (11%). Drugs with the highest efficiency against K. pneumoniae ESBL(+), in our in vitro studies, were: imipenem (100%), meropenem (100%), amikacin (90%) and tetracycline (75%). E. coli ESBL(+) isolates derived from patients of Anesthesiology and Intensive Care Unit (32%), Internal Medicine Unit (16%) and Division of Hematology (13%). Among all tested strains majority were obtained from respiratory tract samples, urine, swabs from wounds and blood (respectively 24%, 24%, 21% and 18%). Isolates of E. coli ESBL(+) demonstrated the greatest susceptibility in case of amikacin (92%) and piperacillin with tazobactam (76%), which suggests the highest activity of that antimicrobials against infections caused by examined strains. None of the analyzed bacilli were resistant to carbapenems. CONCLUSIONS: Our study highlights the importance of characteristics of distribution of ESBL-producing K. pneumoniae and E. coli strains among hospitalized patient. Good antibiotic policies based on antibiotic resistant patterns can decrease the risk of ESBL infection.

[Occurrence of alert pathogens in hospital environment Part II. Multidrug-resistant non-fermenting bacilli]. / Wystepowanie patogenów alarmowych w srodowisku szpitalnym. Czesc II. Wielolekooporne paleczki niefermentujace.

INTRODUCTION: Multidrug-resistant gram-negative non-fermenting bacilli are an important cause of nosocomial infection. Aim of this study was to analyze the prevalence and antimicrobial susceptibility of rods of the species Acinetobacter baumannii and Pseudomonas aeruginosa, belonging to multidrug-resistant alert pathogens. METHODS: 105 (70%) strains of A. baumannii and 46 (30%) strains of P. aeruginosa were isolated from 125 patients hospitalized in the Specialistic Hospital in Krakow, in the years 2008-2010. Taken into account first isolate from the patient. The condition for inclusion in the study was the resistance or reduced susceptibility to selected groups of antibiotics, such as beta-lactams, aminoglycosides and fluoroquinolones. Bacterial identification and antimicrobial susceptibility testing were performed by automated system Vitek 2 Compact (bioMerieux, Poland). All strains were tested with phenotypic method Etest MBL (AB Biodisk, Sweden) for the presence of resistance mechanism associated with the production of metallo-beta-lactamases. RESULTS: Bacilli of the species A. baumannii were isolated most frequently from patients from the Department of Anesthesiology and Intensive Care (52%) and Burn Therapy Unit (25%), with clinical materials collected from the respiratory tract (51%), the wound swabs (18%), urine (11%) and blood (11%). Production of metallo-beta-lactamases was found in 24 (22.9%) strains of A. baumannii. Drugs effective against multidrug-resistant isolates of A. baumannii were colistin and amikacin. Department of anesthesiology and intensive care (59%) and unit of internal medicine (11%) were the main source of multidrug-resistant strains of P. aeruginosa. Pathogens were mainly isolated from clinical specimens collected from the respiratory tract (61%), urine (15%) and wound swabs (13%). Seven (15.2%) strains of P. aeruginosa produced the metallo-beta-lactamases. With regard to colistin and piperacillin with tazobactam was noted the highest percentage of susceptible isolates. CONCLUSIONS: MDR bacteria belonging to alert pathogens are an important cause of many severe and difficult to treat infections which greatly increases the morbidity and mortality among hospitalized patients worldwide. Epidemiological studies and detection of local resistance patterns can provide useful information which can be used in the development of strategies to combat the rising tide of microbial antibiotic resistance.

Distribution of blaOXA genes among carbapenem-resistant Acinetobacter baumannii nosocomial strains in Poland.

Acinetobacter baumannii is an important nosocomial pathogen occurring particularly in intensive care (ICU) as well as burn therapy units (BTU). A. baumannii strains have emerged as resistant to almost all antimicrobial agents, including carbapenems. b-lactamase-mediated resistance is the most common mechanism for carbapenem resistance in this species. Carbapenem-hydrolysing class D b-lactamases - OXA are widespread among A. baumannii strains. It is suggested that ISAba1 plays an important role in drug resistance. The aims of the study were detection of OXA encoding genes and presence of ISAba1. The study included the total of 104 isolates of carbapenem-resistant A. baumannii, obtained from patients hospitalized in ICU and BTU of Specialized Hospital in Krakow. Multiplex PCR was applied for detection of selected OXA carbapenemases encoding genes. PCR analysis showed the presence of bla OXA-51-like gene and ISAba1 in all isolates. 46 strains carried bla OXA-51-like and bla OXA-23-like genes while 48 bla OXA-51-like and bla OXA-40-like genes. 3 isolates carried: bla OXA-51-like , bla OXA-23-like and bla OXA-40-like genes. 7 strains encoded an OXA-51-like carbapenemase but were negative for enzymes belonging to the other families tested. Comparative analysis of ICU and BTU isolates revealed the dominance of: bla OXA-51-like and bla OXA-40-like among ICU while bla OXA-51-like and bla OXA-23-like in BTU.

Variability in Prevalence of Helicobacter pylori Strains Resistant to Clarithromycin and Levofloxacin in Southern Poland.

Background. An increasing resistance of Helicobacter pylori strains to antimicrobial agents is the serious therapeutic problem. The aim of this study was to compare the primary and secondary resistance of H. pylori strains isolated between 2006-2008 (data published) and 2009-2011 to clarithromycin and levofloxacin. Material and Methods. 220 dyspeptic patients (153 before treatment, 67 after), were enrolled in the study. 51 H. pylori strains were isolated. MIC values of clarithromycin and levofloxacin were determined by the E-test method. The statistical analysis was conducted with the χ(2) test with Yates correction at the 0.05 significance level (P ≤ 0.05). Results. Between 2006 and 2008, 34% (39/115) of H. pylori strains were resistant to clarithromycin (primary 21% (19/90), secondary 80% (20/25)). 5% (6/115) of strains were resistant to levofloxacin (primary 2% (2/90), secondary 16% ((4/25); data published) Between 2009-2011, 22% (11/51) of H. pylori strains were resistant to clarithromycin (primary 19% (8/43), secondary 38% (3/8)). 16% (8/51) of strains were resistant to levofloxacin (primary 12% (5/43), secondary 38% (3/8)). Conclusion. The present study has shown the increasing amount of resistant H. pylori strains isolated from patients in Southern Poland to levofloxacin and decreasing number of resistant strains to clarithromycin.