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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 (P0.05). 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 (P0.05). 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.

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.

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.

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.

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.

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.

Summary: The incidence of yeast-like fungi in the genitourinary tract of individuals treated in an out-patient venereal disease clinic was studied. The yeasts occured significantly more rarely in materials taken from patients with acute gonorrhoea in comparison to those without clinical symptoms of gonorrhoea and/or without presence of N. gonorrhoeae in their genitourinary tract. Zusammenfassung: Das Vorkommen hefeahnlicher Pilze im Urogenitaltrakt von Patienten einer venerologischen Poliklinik wurde untersucht. Hefen kamen signifikant sel-tener in Untersuchungsmaterialien von Patienten mit akuter Gonorrhoe vor im Vergleich zu den untersuchten Patienten ohne klinische Symptome einer Gonorrhoe und mit negativem oder positivem Nachweis von Neisseria gonorrhoeae.

Summary: A mycological study of patients attending the dermatological departments in the district of Krakow was undertaken during a period of five years, from 1980 to 1984. The investigations were carried out on 1561 women and 946 men. A total of 3626 specimens (skin scrapings and nails) were investigated. Dermatophytes were isolated from 824 specimens, Candida albicans from 256 and Scopulariopsis brevicaulis from 233. The growth of dermatophytes was observed in 363 samples with negative result of direct microscopic examination. The incidence of mixed fungal cultures from specimens was observed in different combinations. In 8 cases mixed dermatophytes infection took place. Frequently, dermatophytes were isolated accompanied by yeast-like fungi. It should be noted that in 30 samples with positive result of direct microscopic examination, Aspergillus was isolated in 11 cases and S. brevicaulis in 19. Among isolated dermatophytes, T. rubrum was dominant. Zusammenfassung: An Patienten des Distrikts Krakau, welche die Dermatologische Abteilung der Medizinischen Akademie aufsuchten, wurde uber die Funfjahresperiode von 1980 bis 1984 bin eine mykologische Studie durchgefuhrt Die Untersuchungen betrafen 1561 Frauen und 946 Manner. Insgesamt wurden 3626 Materialproben (Hautabschabsel und Nagel) untersucht Aus 824 Proben wurden Dermatophyten, aus 256 Candida albicans und aus 233 Scopulariopsis brevicaulis isoliert Aus 3363 Materialproben mit negativem Ergebnis der direkten mikroskopischen Untersuchung konnten Dermatophyten kultiviert werden. Die Haufigkeit von Pilzmischkulturen in unterschiedlichen Kombinationen wurde bestimmt. In 8 Fallen wurden Mehrfach-Dermatophyten-Infektionen gefunden. Haufig waren Dermatophyten mit Hefen kombiniert Bemerkenswerterweise wurde aus 30 Proben mit positivem mikroskopischem Direktbefund 11 mal Aspergillus und 19 mal Scopulariopsis brevicaulis isoliert. Bei den Dermatophyten herrschte Trichophyton rubrum vor.

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.

Candida albicans frequently inhabits the gastrointestinal tract of humans leading to gastrointestinal candidiasis, especially following suppression of gastric acidity, but studies on the relation between this fungal infection and gastric pathology are limited due to lack of convenient animal models resembling Candida infection in humans. MATERIAL AND METHODS. We compared the effects of C. albicans and vehicle inoculation on gastric secretion and healing of gastric ulcers induced by acetic acid in rats treated with 1) ranitidine (30 mg kg(-1) day(-1) s.c.) and 2) aspirin (ASA) (60 mg kg(-1) day(-1) i.g.) with or without probiotic bacteria Lactobacillus acidophillus. At day 0 and at 4, 15 and 25 days after ulcer induction, the ulcer area, the gastric blood flow (GBF), the quantitative gastric cultures of Candida and the expression of mRNAs for pro-inflammatory cytokines IL-1beta and TNF-alpha and growth factors EGF and TGFalpha were assessed in the gastric mucosa.Gastric acid output was reduced by over 40% soon after Candida inoculation and this effect persisted during all time intervals tested. The area of ulcers in control rats significantly decreased at day 15 and the ulcers disappeared almost completely after 25 days of their induction. In contrast, the ulcers were present until day 25 in Candida-inoculated rats followed by a fall in GBF and a rise in plasma gastrin levels, these effects being significantly attenuated by the co-treatment with Lactobacillus. Candidiasis was accompanied by up-regulation of mRNA for IL-1beta, TNF-alpha, EGF and TGFalpha and a significant increment in plasma IL-1beta and TNF-alpha levels.1) Persistent colonization with Candida could be achieved in rats treated with antisecretory agents or non-steroidal anti-inflammatory drugs (NSAIDs) such as ASA; 2) candidiasis reduces gastric acid secretion, while delaying ulcer healing possibly due to the impairment in GBF in the ulcer area and enhanced expression and release of IL-1beta and TNFalpha and 3) probiotic therapy could be useful in the treatment against the deleterious action of fungal infection on the healing of pre-existing gastric ulcers.

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 Małopolska 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%).

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.

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/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.

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.

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.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.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.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.

Inflammatory bowel diseases (IBD) constitute a group of incurable, inflammatory GI diseases of unknown etiology and the chronic course, with periods of spontaneous disease remissions and aggravations (Muszynski, 2001). This group of diseases includes Crohn`s disease and ulcerative colitis. They are characterized by inflammatory reactions and changes in the structure of intestinal mucosa (Rzeszutko, 2006). IBD etiopathogenesis is not clearly defined. The most essential etiological issues include environmental factors, genetic conditioning, abnormalities of intestinal immunological mechanisms and the bacterial flora (Konturek, 2001). The inflammation which develops in the intestine is mostly due to improper, low-fiber diet which is based on highly processed products and on the poor physical activity. Familial diseases confirmed in 10% of cases speak for the genetic source. Immunological mechanisms which develop in response to food and bacterial antigens are extremely significant (Muszynski, 2001; Polinska et al., 2009). A markedly increased interest has been observed in the intestinal microflora which is believed to be essential in inducing IBD development and in relapses of its clinical symptoms. Bacteria which are normally resident in the GI tract determine the proper work of intestines and protect from excessive proliferation of undesired microorganisms. The contribution of the intestinal flora to the development of the biofilm, which prevents the colonization of pathogens including fungi of the genus Candida in the GI tract, is extremely significant. The area of interest in fungi which are a part of microflora and constitute a potential source of systemic dissemination by colonizing the GI tract has greatly increased lately (Berhardt & Knoke, 1997). In the normal environment fungi are in balance with the bacterial flora and as commensals do not induce the inflammation. The main cause of its development, however, is the immune system impairment and a decrease of immunity. It affects patients with neoplasms and transplant recipients (Pfaller & Diekema, 2007; Warnock & Campbell, 1996). Other diseases causing inflammations are: diabetes and other endocrinopathies, infections due to HIV virus and also surgical procedures, damaged tissues and inflammations of the

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.

The pathogenesis of inflammatory bowel disease (IBD), is related to the effect of fungal presence in the gastrointestinal tract (GI) on IBD development.One hundred twenty-six patients underwent mycological investigation. We analyzed of the occurrence of fungal infection in the clinical material collected from the GI tract of patients with IBD and control groups. We assessed fungal flora transmission from the oral cavity to further segments of the GI tract based on comparative studies of genotypes of fungal strains. The studies were carried out using conventional methods. The fungal genotypes were determined with the PCR-RAPD method.Of 407 samples, 220 of them had fungi. The most frequently isolated species was C. albicans in patients with IBD and in control group, respectively 84.4% and 81%. Analysis of the streak patterns of the products of the PCR-RAPD reaction carried out for C. albicans strains isolated from the material collected from 10 patients, showed 100% affinity between the strains cultivated from the same patient. Genetic affinity between the strains was confirmed in 3 primers used in the examination.The presence of fungi in the oral cavity of patients with IBD may affect more frequent colonization of the colonic mucosa in the active phase of disease. Genetic affinity of C. albicans strains indicates the possibility of fungal transmission from the oral cavity to further segments of the GI tract.

The aim of study was to recognize the epidemiology of hospital acquired pneumonia on the base of genotypes of Acinetobacter baumannii and Pseudomonas aeruginosa strains. Gram negative-non fermenting bacilli are the most frequent aetiologic agent of hospital acquired pneumonia among patients from Anesthesiology and Intensive Care Unit in the Rydygiers hospital in Cracow. In the following research RAPD-PCR method was applied and there 272, 208, ERIC- 2 and PAL- 2 primers were used. The investigation was conducted among 33 Acinetobacter baumannii and 31 Pseudomonas aeruginosa strains which were isolated from endotracheal aspirates (ETA). ETA were taken from 19 patients with microbiologically confirmed pneumonia. The result of our study shows that most A. baumannii strains came from hospital habitat. In the investigated group of A. baumannii strains, 31 belonged to 3 clonal groups and probably came from two bacteria subpopulations which were present in ICU from a long time. There were only two isolates which had got the posthospital origin. Moreover, the assay of genetic similarity between A. baumannii strains, isolated from individual patients, showed that only in two patients were observed strains with different genetic profiles. Isolates which came from 8 patients had got the same pattern of bands. There were many genetic differences between investigated P. aeruginosa strains. In the group of 31 isolates, which were investigated by using 208, 272, ERIC- 2 and PAL- 2 primers, recognized respectively 10, 10, 13 and 8 of genetic profiles. All isolates of P. aeruginosa probably came from a few subpopulations of hospital strain which has undergone evolutionary divergence phenomenon in time as a result of changing condition of hospital environment and application of antibiotics. The assay of genotype of P. aeruginosa strains which were repeatedly isolated from individual patients showed that only from two patients strains with 100% genetic homology were isolated.

Publisher Summary This chapter focuses on the colonization of gastrointestinal (GI) tract by probiotics and fungal. The human GI tract constitutes the second body surface area (250–450 m2). It includes rich microflora of more than 500 species, which makes around 10 microorganisms living in the whole GI tract from the oral cavity area to the colon. These proximal and distal parts of the GI tract are the most densely colonized areas. GI microflora establishes during human life through time-dependent phases. Fungi are a small part of the intestinal microbiota. Many factors such as stress of modern life, reduced physical activity, processed foods, antibiotic therapies, as well as prolonged adaptation of human genes to dynamic changes in the lifestyle seem to derange the host–microflora interplay and contribute to the development of inflammatory and infectious diseases. This process could be attenuated by probiotics. The mechanism that enables probiotics to maintain the balance within the intestine includes production of bacteriocin-like substances, inhibition of pathogens and toxin adherence to the intestinal epithelium, and reduction of the fungal overgrowth.

The aim of the study was the evaluation ofchromogenic agars as a screening media in the routine microbiological diagnostics. 627 clinical samples were cultured on the chromogenic media: chromID S. aureus, chromID MRSA, chromID CPS3 and chromID ESBL from bioMerieux. The results of presumptive identification and detection of selected resistance mechanisms by chromogenic media were compared with results obtained with the conventional methods. Our studies revealed the highest sensitivities of chromID MRSA and chromID CPS3 (90%). The sensivities for chromID ESBL and chromID S. aureus were70%. All media were also highly specific (97%). This specificity for chromID S.aureus and chromID CPS3 was 100%. We conclude, that all tested media showed good performance in presumptive identification and detection of resistance mechanisms.

Nosocomial pneumonia is defined as a pneumonia developing within 48 hours of admission, in a patient with previously normal lung tissue. Ventilator-associated pneumonia (VAP) due to migration of microorganisms from the nose, throat and digestive tract is typical in ITU patients. Microbiological diagnosis is difficult, and there is no single method approved for VAP diagnosis. We have assessed the usefulness of Quantitative Endotracheal Aspirate (QETA) for this purpose, basing on its reported high sensitivity (mean 76% +/- 9%) and specificity (mean 75 +/- 28%).Endotracheal aspirates were sampled from ITU patients fulfilling the following criteria: (1) New or progressive infiltrations in the lungs plus hyperthermia (38.3 degrees C) or hypothermia (36 degrees C); (2) leukocytosis (12 G L(-1)) or leukopenia (4 G L(-1)); or (3) purulent sputum.Nosocomial pneumonia was diagnosed in 40 out of 312 patients treated in the ITU during the study period. VAP was diagnosed in 33 patients. Multi-drug resistant bacteria: Pseudomonas aeruginosa (16), Acinetobacter baumannii (19), ESBL pathogens (8) and MRSA (1) were cultured from QETA in 32 patients. The sensitivity of QETA was 86%.The results allowed for early and accurate diagnosis of nosocomial pneumonia and institution of the best possible treatment.

The aim of study was bacteria characterization, isolated from 30 patients with microbiologically confirmed pneumonia, hospitalized in ICU of Rydygier's Hospital in Krakow. In the study, endotracheal aspirates (ETA) were being collected and quantitatively inoculated. Bacteria isolation at the concentration ofor =10(6) CFU/ml was recognized as bacteriological etiology of pneumonia. Studied 140 clinical materials and isolated 251 bacterial strains from 16 divers species. Isolation of bacteria at the concentration ofor = 10(6) CFU/ml, on the day of clinically recognized HAP, was observed in 26 patients and presence of 1 species of bacteria at the pathological concentration was detected in 17 patients. In the study of endotracheal aspirates, which were collected after 3 and 6 days of treatment, isolation of bacteria species at the significant concentration was observed appropriate in 15 and 10 patients. In the etiology of infections predominated: P. aeruginosa, A. baumannii, S. maltophilia (17 strains; 47.2%) and Enterobacteriaceae bacilli (12 strains; 33%), including bacilli ESBL-positive. The surveillance of bacterial susceptibility patterns detected high percentage of resistance P. aeruginosa strains to imipenem (38.7%).

In the present study on animal model of ulcerative colitis the influence of fungal colonization on the severity of inflammatory lesions in the colon and the course of their healing was evaluated. The results of our studies revealed, that significant fungal colonization (over 10(4) CFU/ml) delayed ulcer healing in the colon. It corresponded with the decrease of colonic blood flow (CBF) in the region of lesions and increase of interleukin (IL)-1beta, tumor necrosis factor(TNF)-alpha level in the serum. Introduction of antifungal therapy (fluconazole) or probiotic in rats inoculated with Candida accelerated the process of ulcer healing in the colon, expressed through the reduction of macro- and microscopic lesions in the colon and decrease of MPO, IL-beta TNF-alpha serum level.

A series of 2-substituted xanthone derivatives 8-20 containing selected allyl, cinnamyl, morpholine, and imidazole moieties were synthesized and tested for their antifungal and antibacterial in-vitro properties. Of the newly synthesized derivatives, ten revealed antifungal activity especially against Trichophyton mentagrophytes (the biggest inhibition zones ranged 35 mm for 11 and 13). 2-(3-(Allylamino)propoxy)-9H-xanthen-9-one hydrochloride 9 inhibited growth of all of the examined fungal species. Significant efficacy against evaluated yeasts and dermatophytes was also observed for 6-chloro-2-methyl-9H-xanthen-9-one derivatives 11-13 containing encyclic amine moieties. Additionally, compounds 9, 11, and 12 hindered development of bacteria species but in a lesser degree. They were efficacious against Staphylococcus aureus, Escherichia coli, and Enterococcus faecalis.

Present-day methods of successful treatment of inflammatory bowel diseases (IBD) result from a better understanding of their pathophysiology due to advances in preclinical studies in this area of knowledge. Until recently microbiological studies have been focused on the bacterial aspects in pathogenesis of GI disorders, however in the last years an interest in the presence of fungi in the gastrointestinal tract has also increased. In this study using an animal model of ulcerative colitis, the impact of fungal colonization of the colon on the intensity of inflammatory changes in the colonic mucosa and the course of their healing was carried out. The macroscopic and microscopic criteria relating to the changes of weight of examined fragments of the colon were evaluated while assessing differences between groups tested. The intensity of intestinal inflammatory changes was determined by assessment of such parameters, as colonic blood flow (CBF), the level of MPO as a marker of colonic neutrophil infiltration intensity and the plasma levels of IL-1beta; and TNF-alpha concentrations. Results at the 3rd day after TNBS rectal administration revealed an increase of weight of isolated segments of inflammed colon, a decrease of CBF and the 4-5 fold increase of plasma MPO activity. Candida colonization of colon mucosa of rats delayed healing of colonic ulcers, induced by TNBS and this was associated with the increased expression of plasma IL-1beta and TNF-alpha levels. Administration of antifungal (fluconazole) or probiotic (Lacidofil) treatment to C. albicans infected rats exerted favorable effect on healing of inflammatory changes in the colon because the area of ulcerations in groups of rats treated with fluconazole or Lacidofil was significantly smaller in comparison with those inoculated with Candida solution only. Administration of fluconazole or Lacidofil significantly decreased the weight of colon segments, the MPO activity and the plasma IL-1beta and TNF-alpha levels, as compared with respective values in the group receiving Candida only. The results of our studies indicate the deteriorating influence of Candida on the healing process of inflamed colon in the animal model of ulcerative colitis. Concomitant therapy with probiotic or antifungal treatment improved healing of colonic lesions, decreased the weight of inflamed colonic tissue and also attenuated the MPO activity and plasma proinflammatory cytokines IL-1beta and TNF-alpha levels.

The aim of the study was evaluation of susceptibility of Pseudomonas aeruginosa strains isolated from patients hospitalized in different wards of Rydygier's Hospital in Krakow in 2005. Bacteria were identified on the basis of typical morphology confirmed by Gram-staining microscopy and by biochemical tests--ID 32 GN strips using ATB system (bioMerieux, France). The susceptibility of all isolates to a panel of antimicrobial agents was performed using disk diffusion method. The highest in vitro activity against clinical strains demonstrated ceftazidime (88.6% of susceptible strains) while the lowest in vitro activity against clinical strains demonstrated imipenem (50.4% of susceptible strains). It was also observed that 40.2%(53) of strains were resistant to meropenem and imipenem. Carbapenem resistant P aeruginosa strains were tested for MBL production. We performed disk synergy test for MBL detection with EDTA, 2-MPA and ceftazidime, imipenem. The presence of a distorted inhibition zone was interpreted as a positive result for MBL production. Positive results of disk synergy tests were confirmed by Etest MBL strips. Metalo-beta-lactamases were detected in 13 isolates resistant to carbapenems.

The influence of microbiological factor is taken into consideration in the pathogenesis of ulcerative colitis (UC). The aim of studies was to: 1.evaluate the presence of significant fungal colonization, over 10(5) CFU/ml in patients with UC and the control group (irritable colon syndrome, IBS); 2. estimate the influence of antifungal treatment in the activity of UC.We evaluated 72 patients aged from 18-72 years, 60 patients with UC, 12 with IBS. Clinical investigation: initially and after 4 weeks interview and colonoscopy with colon biopsies for histology and mycology were taken. Activity of UC was evaluated according to: clinical, endoscopic and histological IACH criteria. 13 patients with significant fungal colonization were given antifungal treatment. Biopsies for histology were stained with hematoxylin-epsin (H-E). Qualitative and quantitative mycolo-gical evaluation was performed according to Muller method.1. Significant fungal colonization was more frequent in patients with UC history over 5 years, in comparison with shorter disease history and IBS, in 33.3%, 13.8% and 1.3% respectively. 2. Candida albicans was most often isolated in 91.7% of cases. 3. Initial analysis of the activity index of UC in patients with significant and non-significant fungal colonization did not revealed differences between these groups, 13.84 and 14.0 respectively. 3. After 4 weeks stronger decrease of the UC activity index was observed in patients with significant fungal colonization treated with antifungal treatment, in comparison with patients not given antifungal therapy: 8.0 and 10.41 respectively, p0.01. Differences were significant according to clinical 2.23 (C) -3.33 (D), p0.05 and endoscopic cryteria: 3.46 (C) -4.84 (D), p0.01.1. Significant fungal colonization of colon may influence the activation of UC. 2. Longer disease history may be the risk factor of significant fungal colonization in colon. 3. Antifungal treatment in patients with significant colonization caused clinical improvement of UC.

The aim of our investigations was the microbiological analysis together with the evaluation of sensitivity of bacteria frequently isolated from blood cultures. Blood samples were taken from patients with symptoms suggesting bacteremia in Rydygier's Hospital in Cracow. A total of 11,170 blood samples taken from 1997 to 2000 were tested. Automatic VITAL system (bioMerieux) was applied to culture and detect microorganisms. Bacteria were identified by ATB system (bioMerieux). Susceptibility was detected by ATB and disc diffusion method. Percentage of positive results relating to detection of microorganisms of clinical significance was 16.9% (1891 cultures). Staphylococcus spp. (Staph. epidermidis in range 22.8% to 21.9%), Enterococcus spp. and Streptococcus spp. were most frequently isolated species among aerobic Gram-positive bacteria. In 2000, compared to 1997 the number of isolates of MRSA increased considerably (from 1.8% to 6.8%). In blood infections the increase of frequency of E. coli bacteria was also noted: 6.1% and 11.4% in 1997 and 2000, respectively. Among non-fermentant bacilli the percentage of occurrence of P. aeruginosa in the period of 4 years was comparable in the range 7.3% in 1997 to 7.2% in 2000. The increase in the frequency of blood infections of A. baumanii was also noticed (respectively from 4.8% to 9.9%). Susceptibility of P. aeruginosa strains to selected beta-lactame antibiotics and aminoglycosydes increased in 2000 in comparison to 1999. A. baumanii strains were 100% sensitive only to imipenem.

The aim of this study was to estimate the frequency of fungal colonization of the stomach of patients suffering from gastric ulcer (GU) and chronic gastritis (CG) and the influence of fungal colonization of the stomach on the process of ulcer healing. We investigated 293 patients aged 20-80 years. Before and after 4 weeks of sucralfate treatment they underwent endoscopy of the stomach, histological examination of biopsies taken from the ulcer margin or inflamed gastric mucosa and mycological examinations of the gastric juice, surface brushing and biopsies. The studies revealed a high concentration of fungi in 54.2% patients with GU and 10.3% with CG. Candida albicans was the most frequently isolated organism. Fungal colonization of the stomach impairs the process of gastric ulcer healing. Control examination after 4 weeks of sucralfate therapy showed the ratio of GU healing in 62% of patients with a high concentration of fungi in comparison with 78% of patients not colonized with fungi (P < 0.05). A significantly longer duration of ulcer symptoms in the group of patients with a high concentration of fungi in the stomach was also observed. There was no correlation between the level of fungal antibodies, of Candida antigen in the serum and the concentration of fungi in the stomach.

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.