Fecal microbiota transplantation - methods of treatment of recurrent Clostridium difficile infections and other diseases.

Clostridium difficile is a serious epidemiological problem and particularly dangerous microorganism causing hospital infections. Currently, the treatment of C. difficile infections is the use of metronidazole or vancomycin. However, in some patients, recurrent infection difficult to treat occurs. Fecal microbiota transplantation (FMT) is a new method used to treat the recurrent CDI. FMT consists in the infusion of the fecal suspension from a healthy donor into the gastrointestinal tract of a patient with CDI to restore the natural intestinal microflora. FMT is safe and effective treatment of recurrent CDI. FMT is extensively described around the world, but to date only two randomized studies confirming the effectiveness of FMT have been conducted. This method was also applied in the treatment of diseases such as pseudomembranous colitis, ulcerative colitis, Crohn's disease and irritable bowel syndrome. The review describes the procedure for FMT and the current state of knowledge about the effectiveness of FMT in the treatment of recurrent CDI.

The use of the HRM method for identifying possible mutations in the ymoA gene region and evaluating their influence on the enterotoxic properties of Y. enterocolitica strains.

BACKGROUND: The yst gene that encodes the production of Yst enterotoxins is one of the most important and reliable virulence markers. Its ability to produce Yst has been demonstrated in pathogenic strains isolated from clinical cases of yersiniosis with diarrhea. However, not all yst positive strains produce enterotoxins. According to some authors, Yst production can be restored in a silent strain by ymoA mutation. In this study, the HRM method was applied to identify ymoA single nucleotide polymorphism with the aim of evaluating their influence on the enterotoxic properties of Y. enterocolitica strains. RESULTS: Two genotypes (A and G) of the examined nucleotide sequence and some variations were detected in the HRM analysis. A phylogenetic analysis of 10 genotype A nucleotide sequences revealed 100% similarity with the Yersinia enterocolitica subsp. enterocolitica 8081 genome NCBI Acc. No. AM286415. An analysis of 10 genotype G nucleotide sequences and 3 variations sequences revealed two point mutations in the examined region: transition A3387326G and insertion A in position 3387368. However, no mutations were observed in the coding region of any of the examined ymoA gene fragments. Genotype G was identified in nearly all Y. enterocolitica strains isolated from pigs. Only 4 nucleotide sequences were similar to AM286415 and did not feature point mutations. In case of human Y. enterocolitica strains 31 were classified as belonging to genotype A, the remaining 59 belonged to genotype G and were characterized by the presence of point mutations. CONCLUSIONS: No correlations were observed between enterotoxic properties and the presence of mutations in the ymoA gene region of Y. enterocolitica strains isolated from both humans and pigs.

In vitro effect of amphotericin B on Candida albicans, Candida glabrata and Candida parapsilosis biofilm formation.

Candida spp. biofilm is considered highly resistant to conventional antifungals. The aim of this study was to investigate the in vitro effect of amphotericin B on Candida spp. biofilms at different stages of maturation. We investigated the activity of amphotericin B against 78 clinical isolates of Candida spp., representing three species, growing as planktonic and sessile cells, by a widely accepted broth microdilution method. The in vitro effect on sessile cell viability was evaluated by MTT reduction assay. All examined strains were susceptible to amphotericin B when grown as free-living cells. At the early stages of biofilm maturation 96.7-100.0 % strains, depending on species, displayed amphotericin B sessile minimal inhibitory concentration (SMIC) ≤ 1 µg/mL. Mature Candida spp. biofilm of 32.1-90.0 % strains displayed amphotericin B SMIC ≤ 1 µg/mL. Based on these results, amphotericin B displays species- and strain-depending activity against Candida spp. biofilms.

Comparison of detection methods for extended-spectrum beta-lactamases in Escherichia coli strains.

INTRODUCTION: Detection of extended-spectrum beta-lactamases (ESBLs) could be a major challenge for microbiologists--the difficulties arise mainly from the phenotypic differences among strains. MATERIALS AND METHODS: Evaluation of ESBLs was performed on 42 strains of E. coli by: 1) DDST on MHA, 2) DDST on MHA with cloxacillin, 3) CT on MHA, according to CLSI, 4) CT on MHA with cloxacillin, 5) Etest ESBL (AB Biodisk), 6) CHROMagarTM ESBL (GRASO), 7) ChromID® ESBL (bioMérieux), and 8) automatic system VITEK2 ESBL test (bioMérieux). RESULT: Positive results were obtained for 20 strains using method 1, for 18 strains using method 2, 17 by method 3, 14 by method 4, 11 by method 5, 39 by method 6, 40 by method 7, and 15 by method 8. Using Etest ESBL 6.0 non-determinable results were obtained. The most consistent results were obtained when comparing the results of method 3 with results of method 2 (97.6%), and comparing the results obtained using methods 3 and 8 (95.2%). CONCLUSIONS: Based on our study we conclude that the chromogenic media can only be used as a screening method for the detection of ESBLs in E. coli rods. Etest is less useful compared to other phenotype methods, due to the impossibility of obtaining results for all the tested strains. Adding cloxacillin to MHA does not increase the frequency of detection of ESBLs in E. coli strains. DDST seems to be the most reliable among phenotypic methods for the detection of ESBLs in E. coli rods.

Candidaemia in polish hospitals - a multicentre survey.

Significant changes in the frequency of candidaemia and the distribution of causative species have been noted worldwide in the last two decades. In this study, we present the results of the first multicentre survey of fungaemia in Polish hospitals. A total of 302 candidaemia episodes in 294 patients were identified in 20 hospitals during a 2-year period. The highest number of infections was found in intensive care (30.8%) and surgical (29.5%) units, followed by haematological (15.9%), 'others' (19.2%) and neonatological (4.6%) units. Candida albicans was isolated from 50.96% of episodes; its prevalence was higher in intensive care unit and neonatology (61.22% and 73.33%, respectively), and significantly lower in haematology (22%; P < 0.001). The frequency of C. krusei and C. tropicalis was significantly higher (24% and 18%) in haematology (P < 0.02); whereas, the distribution of C. glabrata (14.1%) and C. parapsilosis (13.1%) did not possess statistically significant differences between compared departments. Obtained data indicates that species distribution of Candida blood isolates in Polish hospitals reflects worldwide trends, particularly a decrease in the prevalence of infections due to C. albicans.

[The evaluation of lipolytic activity of strains of Enterococcus faecium]. / Ocena aktywnosci lipolitycznej szczepów Enterococcus faecium.

INTRODUCTION: Little is known about the involvement of hydrolytic enzymes such as lipases as virulence factors in infections involving Enterococcus spp. METHODS: A total of 45 isolates of E. faecium were investigated. Lipolytic activity of enterococcal strains was determined by Tryptic Soy Agar containing Tween 20, Tween 40, Tween 60, Tween 80 and egg yolk. RESULTS: We detected that E. fecium strains produced lipases more frequently on Tween 20 agar (71.1% strains) than on Tween 40 agar, Tween 60 agar, Tween 80 agar, egg yolk agar (respectively 33.3%, 24.4%, 20.0%, 31.1%). Our results indicate that lipase may be a virulence factor in E. faecium. CONCLUSIONS: Studies suggest that source of isolation from clinical materials (blood, wound and fluid from the abdominal cavity) does not have an influence on the ability hydrolysis esters.

[Evaluation of selected properties of morphological variants of Acinetobacter baumannii complex]. / Ocena wybranych wlasciwosci morfologicznych wariantów paleczek Acinetobacter baumannii complex.

INTRODUCTION: The ability of growth of Acinetobacter baumannii as morphology colony variants have been observed. However, the importance of this phenomenon for its biology is not known. The aim of this study was to evaluate some properties of light and dark morphology colony variants. METHODS: Fifty two isolates were identified by MALDI TOF MS method (MALDI Biotyper, BRUKER). It was evaluated the adhesion to polystyrene and extracellular mucus production of morphology colony variants and its susceptibility to imipenem and meropenem by agar dilution method. RESULTS: Forty eight (92.3%) out of the 52 morphotypes Acinetobacter sp. were identified as A. baumannii, two (5.8%) as A. genomospecies 3, one as the A. calcoaceticus. Sixteen (61.0%) pairs of isolates showed differences in the similarity of the spectra to the spectra of reference strains in the MALDI-TOF MS method. Adhesion to polystyrene was observed in all dark and 92.3% light morphotypes. Extracellular slime was produced by 15 (57.7%) dark morphotypes, and 7 (26.9%) of clear. The differences in susceptibility to imipenem occurred in two (7.7%), and meropenem in three (11.5%) pairs of morphotypes. CONCLUSIONS: The results show diversity of biological properties of morphology colony variants of A. baumannii complex. Differences in the level of adhesion to polystyrene and slime production may indicate the importance of morphological differentiation in virulence of A. baumannii complex.

Does Toxoplasma gondii infection affect cognitive function? A case control study.

According to studies, latent Toxoplasma gondii infection may affect several functions of the human brain. Here we search for the association between latent toxoplasmosis and cognitive performance. We tested 70 individuals for latent T. gondii infection. There were 26 Toxoplasma-infected subjects and 44 Toxoplasma-free subjects. Within these two groups we assessed cognitive performance using a set of standardized, widely recognized neuropsychological tests: Trail Making Test, Stroop Test, Verbal Fluency Test, Digit Span Test and N-back test. The relationship between chronic toxoplasmosis and cognitive performance was assessed, with adjustment for age and sex. Patients with latent toxoplasmosis performed worse on one neuropsychological test, N-back Test--percentage of correct answers (beta -8.08; 95% CI - 15.64 to -0.53; p < 0.05) compared to seronegative patients. However, after adjustment for age and sex, no statistically significant associations between latent toxoplasmosis and the scores on any cognitive tests were noticed. As statistically significant relationship was not observed, this study does not confirm that chronic latent T. gondii infection affects cognition.

Nosocomial outbreak due to Klebsiella pneumoniae at a neurosurgery unit.

Thirty three isolates of K. pneumoniae were studied. The strains were cultured from different clinical specimens received from patients hospitalised at a Neurosurgery Unit at the Dr Jurasz University Hospital in Bydgoszcz. Production of ESBL was assessed using double disk synergy test. The genomic DNA was extracted from the strains separated by PFGE after digesting with XbaI endonuclease. Production of ESBL was detected in 81.8% of K. pneumoniae isolates. Molecular typing results revealed a great genetic diversity among K. pneumoniae isolates. All repeated PFGE patterns were detected in 12 (36.3%) K. pneumoniae isolates.

[Morganella sp. rods--characteristics, infections, mechanisms of resistance to antibiotics]. / Paleczki Morganella--charakterystyka, zakazenia, mechanizmy opornosci na antybiotyki.

The Morganella genus is one member of the tribe Proteae, which also includes the genera Proteus and Providencia. These bacteria are commonly present in the environment. Morganella sp. rods are known to be a causative agent of opportunistic hospital infections, mainly urinary tract, wound and blood infections of severe and high mortality, even in cases of an appropriate antibiotic. These bacteria may produce many virulence factors, for example urease, hemolysins, LPS, adhesins and enzymes hydrolyzing and modifying antibiotics commonly used to treat infections. Understanding the diverse biological properties of these rods may be of importance in the development of effective methods of prevention and control of infections with their participation.  

[Adhesion and slime production by wild and clinical strains of Acinetobacter baumannii]. / Adhezja oraz wytwarzanie sluzu zewnatrzkomórkowego przez dzikie i kliniczne szczepy paleczek Acinetobacter baumannii.

INTRODUCTION: Adhesion of bacteria to the surface plays a key role in the development of infection, and is the first stage of biofilm formation. The ability of A. baumannii strains to adhesion and forming biofilms on abiotic surfaces, as well as eucaryotic cells was described.A. baumannii is also capable of secretion of the exopolysaccharide (EPS) - a substance that allows the binding of bacterial cells to the surface, and with each other. The aim of this study was to evaluate the ability of biofilm formation and slime production by wild-type and clinical strains of A. baumannii. METHODS: We examinated 51 strains ofA. baumannii, including 14 isolated from lower respiratory tract, 17 from wound swabs and 20 from the soil. Adhesion to polystyrene was evaluated by modified Christensen methods and slime production by Ishiguro method. RESULTS: Adhesion to polystyrene was observed in 51,0% of strains, including 70,0% of wild-type and 38,7% of clinical strains (64,7% strains from wound swabs and one strain from lower respiratory tract). Slime production was found in 31,4% of strains, of which the largest (42.9%) group strains were isolated from lower respiratory tract. There was no correlation between production of extracellular slime, and the adhesion of strains to polystyrene. CONCLUSIONS: Different levels of expression of virulence factors in A. baumannii strains isolated from different origin indicates their importance in the colonisation ecological niches and the development of infections at various sites.

[Occurrence and antimicrobial susceptibility of Morganella morganii strains isolated from clinical samples]. / Wystepowanie oraz lekowrazliwosc szczepów Morganella morganiiizolowanych z materialu klinicznego.

The aim of this study was the evaluation of occurrence and antimicrobial susceptibility of M morganii rods isolated from clinical samples. This study included 201 strains isolated in the Clinical Microbiology Department of Dr. A. Jurasz University Hospital in 2008-2010. Identification to species was carried out on the basis of the results of biochemical reactions included in the tests ID 32E and VITEK2 GN. Antimicrobial susceptibility of M. morganii rods was determined by the disk-diffusion method on Mueller-Hinton II Agar. Strains of M morganii most commonly isolated from skin and soft tissue, and material taken from the urinary tract, mainly from patients of Anesthesiology and Intensive Care Unit, Department of General and Vascular Surgery and Department of General Surgery and Endocrinology. All of M morganii strains isolated during the three years were susceptible to carbapenems. We reported decrease of strains susceptible to piperacillin and chloramphenicol. In 2010 we showed a higher percentage of strains intermediate to tigecycline, compared with 2009. We observed increase in the percentage of strains resistant to cefoperazone with sulbactam and reported decrease in the percentage of strains resistant and intermediate to aminoglycosides. Extended Spectrum Beta-Lactamases were produced by 13 (6,5%) of M morganii strains.

Influence of subinhibitory concentrations of cefotaxime, imipenem and ciprofloxacin on adhesion of Escherichia coli strains to polystyrene.

The present study investigated the ability of sub MICs of cefotaxime, imipenem and ciprofloxacin to interfere with adhesion of E. coli strains to polystyrene (selected polymer used in studies on microorganisms' adhesion). It was observed that cefotaxime and imipenem at 1/2 and 1/4 MICs decreased the adherence of E. coli strains to polystyrene significantly. 1/2, 1/4 and 1/8 MICs of ciprofloxacin generally decreased the adhesive properties of E. coli strains, but two E. coli strains showed a noticeable enhancement of adhesion after incubation at sub MICs of this antibiotic.

[Susceptibility of Klebsiella pneumoniae isolated from urinary tract infections to quinolones]. / Wrazliwosc na fluorochinolony paleczek Klebsiella pneumoniae izolowanych z moczu.

Urinary Tract Infections (UTIs) are the most prevalent infections worldwide both in males and females. K. pneumoniae is one of the major pathogens causing UTIs. Fluoroquinolones are effective drugs for treating infections caused by Klebsiella spp. The aim of this study was to evaluated the susceptibility to three fluoroquinolones of K. pneumoniae strains isolated from urine. The MICs of ciprofloxacin was determined by agar dilution method and the MICs of norfloxacin and gatifloxacin by E-test. Among analysed K. pneumoniae strains 86.7% was susceptible to gatifloxacine, 76.7% to norfloxacin and 51.2% to ciprofloxacine.

[Susceptibility of Klebsiella oxytoca to selected antibiotics]. / Wrazliwosc paleczek Klebsiella oxytoca na wybrane antybiotyki.

Fifty two clinical isolates of K. oxytoca were included. All of analysed strains were isolated from wound swabs. The aim of this study was to evaluate MIC value of amoxicillin with clavulanic acid, tigecycline and ciprofloxacin. The susceptibility to amoxicillin with clavulanic acid and tigecycline was tested by the Etest. The susceptibility to ciprofloxacine was tested by the agar dilution method. Among of analysed K. oxytoca strains 44 (84.6%) were susceptible to tigecycline, 27 (51.9%) to amoxicilline with clavulanic acid and 21 (40.4%) to ciprofloxacine. These data suggest that tigecycline, may be an effective therapeutic option for the treatment infections caused by K. oxytoca strains.

[High level of aminoglycoside resistance among Enterococcus faecalis and Enterococcus faecium strains]. / Opornosc na wysokie stezenia aminoglikozydów wsród szczepów Enterococcus faecalis i Enterococcus faecium wyosobnionych z materialu klinicznego.

Enterococcus sp. strains are believed as important reason of serious nosocomial infections currently. These infections are cured by using combination of beta-lactams and aminoglycosides for their treatment. Enterococcus sp. resistant to high-level doses of aminoglycosides, beta-lactams and vancomycin are responsible for therapeutic failure. The aim of our study was to evaluate the incidence of isolation and susceptibility to antibiotics of HLAR Enterococcus sp. strains isolated between 2007 and 2010 from the patients of University Hospital No. 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz Nicolaus Copernicus University in Torun. Amongst 6137 Enterococcus sp. strains 1124 (18,3%) presented HLAR phenotype; 53,1% of them was identified as E. faecalis and 46,9% as E. faecium. The highest percentage of all examined strains was isolated from the patients of different surgery clinics, Intensive Care Units, and Pediatrics, Hematology and Oncology Clinic. HLAR and HLSR phenotypes were noted in E. faecalis, for 45,7% and 27,5% strains, in E. faecium - 29,8% and 9,5%, respectively. HLGR phenotype was presented twice more often in E. faecium than E. faecalis. Highest percentages of E. faecium resistant to glycopeptides and rifampicin were observed when compared with E. faecalis. The highest percentages of strains intermediate, resistant to vancomycin and resistant to glycopeptides were noted for E. faecium strains with phenotypes HLAR, HLGR and HLSR.

[Antimicrobial sensitivity of Escherichia coli strains with K1 antigen isolated from pregnant women and newborns]. / Lekowrazliwosc szczepów Escherichia coli z antygenem K1 izolowanych od kobiet ciezarnych i noworodków.

The aim of this study was comparison of the susceptibility to antibiotics of E. coli strains with K1 antigen (E. coli K1+) and non-K1 E. coli strains (E. coli K1-). This study included 67 of E. coli K1+ and 67 of E. coli K1- strains isolated in the time period from June to September of 2008 from pregnant women and newborns hospitalized at dr. J. Biziel University Hospital number 2 L. Rydygier Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Torun. Antimicrobial susceptibility of E. coli strains was tested by the disc-diffusion method, on the Mueller Hinton 2 Agar (Becton Dickinson). It was found that 64,2% of E. coli K1+ strains and 53,7% of E. coli K1-strains were susceptible to all tested antibiotics and chemioterapeutics. E. coli K1- strains were more often than E. coli K1+ nonsusceptible to at least one antimicrobial agent. The obtained results indicate that E. coli K1+ strains significant differed in the susceptibility to ampicillin/sulbactam (85,1% versus 95,5%) (p=0,041), cephalothin (70,1% versus 85,1%) (p=0,038) and tetracycline (91,0% versus 74,6%) (p=0,012) from E. coli K1-strains. All tested E. coli K1+ and K1-strains were sensitive to piperacillin/tazobactam, cefoperazone/sulbactam, cefotaxime, ceftazidime, cefepime, imipenem, amikacin, netilmicin and tigecycline. There weren't the ESBL-producing strains among tested E. coli K1+ and K1- rods.

[Evaluation of biofilm formation by Proteus mirabilis strains on the surface of different biomaterials by two methods]. / Porównanie dwiema metodami tworzenia biofilmu przez paleczki Proteus mirabilis na powierzchni róznych biomaterialów.

Proteus sp. rods are opportunistic human pathogens. These microorganisms are mainly isolated from patients with urinary tract infections, particularly associated with using of biomaterials, on which surface they can form biofilm. The aim of our study was the estimation of Proteus mirabilis rods ability to form biofilm on the surface of 5 biomaterials (polychloride vinyl, silicone latex, polypropylene, polybutylen teraftalan and polyamide) using Richards' and quantitative method and comparison results of both methods. A total number of 84 P. mirabilis strains were included into the study. All of them were isolated in the Department of Clinical Microbiology University Hospital no. 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz, Nicolaus Copernicus University in Torun between 2005 and 2008. Examined P. mirabilis strains formed heavy biofilm with statistically significantly values on the surface of silicone latex than on polychloride vinyl and on polypropylene surface than polybutylen teraftalen or polyamide. High correlation of both methods was established. The Richards' method can be used to quick identification of P. mirabilis biofilm.

[The evaluation of amylolytic activity of strains of coagulase negative staphylococci]. / Ocena amylolitycznej aktywnosci szczepów gronkowców koagulazo-ujemnych.

Amylase activity of 30 strains of Staphylococcus spp. was determined by Tryptic Soy Agar on supplemented with 1.0% starch as the substrate. After incubation (time incubation 24 h or 168 h), the plates were flooded with Lugol solution. A clear zone around the colonies indicated amylase activity. The 23 (76.7%) strains CNS demonstrated the amylase activity. It was observed that 17 (80.9%) strains of S. epidermidis, and 6 (66.7%) strains non-S. epidermidis, starch hydrolyzed. Amylase production depends of time incubation (frequently 168 h) and growth atmosphere (frequently oxygen atmosphere)

[Susceptibility to antibiotics of Enterobacter spp. rods isolated from urine]. / Ocena lekowrazliwosci szczepów enterobacter sp. izolowanych z moczu.

Enterobacter spp. rods are opportunistic microorganisms which cause of urinary tract infections. The aim of this study was the evaluation of the susceptibility to antimicrobial agents antibiotics of Enterobacter spp. rods isolated from urine. The study was carried 50 of Enterobacter spp strains isolated in the Clinical Microbiology Department of dr. A. Jurasz University Hospital. Antibiotic susceptibility was tested by disk diffusion method. All of strains were susceptible to imipenem and meropenem. There was 87,5% of strains sensitive to doripenem, 79,2% to ertapenem, 54,0% to piperacillin/tazobactam and 50,0% to cephepime. The relatively high percentage (62,0%) of Enterobacter spp. was sensitive to fluoroquinolones. Extended spectrum beta-lactamases were produced by 24 (48,0%) strains.