Eugenia Gospodarek

Reduced expression of virulence factors in multidrug-resistant Pseudomonas aeruginosa strains

MDR Pseudomonas aeruginosa strains are isolated from clinical specimens with increasing frequency. It seems that acquiring genes which determine antibiotic resistance usually comes at a biological cost of impaired bacterial physiology. There is no information on investigations comparing phenotypic differences in MDR and MDS P. aeruginosa strains in literature. The study included 150 clinical P. aeruginosa isolates (75 classified as MDS and 75 as MDR). PFGE analysis revealed five pairs of identical isolates in the group of MDR strains and the results obtained for these strains were not included in the statistical analyses. MDR strains adhered to polystyrene to a lesser extent than MDS strains. The growth rate in the liquid medium was significantly lower for MDR strains. Detectable amounts of alginate were present in the culture supernatants of seven MDS and six MDR strains. The MDR P. aeruginosa strains which were investigated produced significantly lower amounts of extracellular material binding Congo Red, lower lipolytic, elastase, LasA protease, phospholipase C activity and pyocyanin quantity in culture supernatants when compared with MDS strains. No significant differences were observed between MDR and MDS strains in proteolytic activity. In conclusion, the MDR P. aeruginosa strains have impaired virulence when compared to MDS strains.

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.

Human meningitis caused by Streptococcus suis

Streptococcus suis is an important swine pathogen worldwide, which can be transmitted to human beings by direct contact; therefore, S. suis infections occur mainly in people who handle pigs or pork. We present a case of a patient with S. suis meningitis who worked as a butcher in a meat processing plant for 5 years. The 35-year-old man was admitted to the Department of Infectious Diseases in T. Browicz Memorial Central Infectious Disease and Observation Hospital in Bydgoszcz, Poland, with suspected bacterial meningitis. According to his medical history, the patient had been injured during the processing of pork. A microbiological examination of the cerebrospinal fluid and blood revealed S. suis as a single aetiological factor of this infection. The patient was empirically administered cefotaxime (2.0 g at 8-h intervals) and penicillin (9 million U at 8-h intervals). The patient made a complete recovery and his inflammatory markers normalized. Only the hearing deficit of his right ear did not disappear. An otolaryngologist recommended a 4-week steroid therapy. The patient was not examined because he did not report to the clinic. To our knowledge this is the first described case of human meningitis caused by S. suis in Poland.

Prevalence of genes encoding virulence factors among Escherichia coli with K1 antigen and non-K1 E. coli strains

Multiplex PCR was used to detect genes encoding selected virulence determinants associated with strains of Escherichia coli with K1 antigen (K1(+)) and non-K1 E. coli (K1(-)). The prevalence of the fimA, fimH, sfa/foc, ibeA, iutA and hlyF genes was studied for 134 (67 K1(+) and 67 K1(-)) E. coli strains isolated from pregnant women and neonates. The fimA gene was present in 83.6 % of E. coli K1(+) and in 86.6 % of E. coli K1(-) strains. The fimH gene was present in all tested E. coli K1(+) strains and in 97.0 % of non-K1 strains. E. coli K1(+) strains were significantly more likely to possess the following genes than E. coli K1(-) strains: sfa/foc (37.3 vs 16.4 %, P = 0.006), ibeA (35.8 vs 4.5 %, P<0.001), iutA (82.1 vs 35.8 %, P<0.001) and hlyF (28.4 vs 6.0 %, P<0.001). In conclusion, E. coli K1(+) seems to be more virulent than E. coli K1(-) strains in developing severe infections, thereby increasing possible sepsis or neonatal bacterial meningitis.

Toxin A-producing Clostridium difficile as an aetiological factor of post-traumatic wound infection

Clostridium difficile is a well-known cause of hospital-acquired infection such as antibiotic associated diarrhoea or pseudomembranous colitis. Extraintestinal infections caused by this pathogen are described rarely. A case of post-traumatic wound infection caused by C. difficile in an immunocompetent, young and otherwise healthy trauma patient is reported. A 31-year-old female, a car accident victim, was admitted to hospital because of polytrauma. After open reduction and internal fixation of a supracondylar femoral fracture by means of the dynamic condylar screw (DCS) system, a purulent fistula occurred. Microbiological examination of the pus revealed C. difficile as the single aetiological factor of this infection. Empirical antibiotic treatment with cefazoline and metronidazole had been administered right after the surgery, but was found to be ineffective. The strain isolated from the patient was sensitive to most antimicrobials except for clindamycin, and amoxicillin/clavulanic acid was chosen for the guided therapy. Such treatment combined with the removal of the DCS system produced a desirable effect.

Catheter-related blood stream infection caused by Raoultella ornithinolytica.

Raoultella spp. representatives are Gram-negative capsulated, nonmotile rods. These bacteria are found in the natural environment: plants, water, soil and insects. R. ornithinolytica is one of the three species of Raoultella. R. ornithinolytica is the only species within the genus which has the ability to produce ornithine decarboxylase. Human infections related to R. ornithinolytica are exceedingly rare. The present case report describes catheter-related blood stream infection caused by R. ornithinolytica and successfully treated with antibiotic therapy.

Antimicrobial Activity and SAR Study of New Gemini Imidazolium-Based Chlorides

A series of 70 new 3,3′(α,ω‐dioxaalkyl)bis(1‐alkylimidazolium) chlorides were synthesized. They were characterized with respect to surface active properties and antimicrobial activity against the following pathogens: Staphylococcus aureus, Enterococcus faecalis, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, Candida krusei, and Candida albicans. In this article, besides description of the synthesis, we characterize a set of features of these compounds, concerning their structure (described by the length of the dioxaalkan spacer and the length of the alkyl substituent in the aromatic ring) and surface active properties (critical micelle concentration, value of surface tension at critical micelle concentration, value of surface excess, molecular area of a single particle, and free energy of adsorption of molecule). Then, we present a SAR study for Staphylococcus aureus, as one of the most widespread pathogenic strains, conducted with the help of the Dominance‐based Rough Set Approach (DRSA), that involves identification of relevant features and relevant combinations of features being in strong relationship with a high antimicrobial activity of the compounds. The SAR study shows, moreover, that the antimicrobial activity is dependent on the type of substituents and their position at the chloride moiety, as well as on the surface active properties of the compounds.

Detection of Helicobacter pylori and cagA gene in nasal polyps and benign laryngeal diseases.

BACKGROUND AND AIMS Helicobacter pylori is the most common etiological factor of chronic infection worldwide. It has also been found in human dental plaques, mouth, saliva, tonsils and adenoid tissue, medial ear or nasal polyps and sinuses mucosa, as well in several benign and malignant lesions of the larynx and pharynx. The aim of the study was to investigate the association of H. pylori colonization in chronic rhinosinusitis and benign laryngeal diseases. METHODS The prospective, controlled study involved a series of 30 patients with nasal polyps and normal nasal mucosa and 30 patients with benign laryngeal diseases. Samples of 10-15 mg obtained from fresh tissues were used for nucleic acid purification. All samples were subjected to H. pylori ureA detection by the PCR H. pylori diagnostic test. Samples that were positive for ureA H. pylori gene were evaluated for cagA H. pylori gene. RESULTS H. pylori DNA (ureA gene) was detected in all patients with nasal polyps, concha bullosa and laryngeal diseases. Presence of H. pylori cagA gene was identified in 7 (23.3%) of 30 patients of H. pylori-positive larynx samples and no positive result was observed in nasal polyps and concha bullosa. CONCLUSIONS Our results reveal the presence of H. pylori DNA in nasal polyps, concha bullosa and benign larynx diseases. cagA-positive H. pylori was observed only in laryngeal tissues. These results may have implications for a possible role of H. pylori in laryngeal diseases.

The Assessment of Proteus mirabilis Susceptibility to Ceftazidime and Ciprofloxacin and the Impact of These Antibiotics at Subinhibitory Concentrations on Proteus mirabilis Biofilms

Rods of the Proteus genus are commonly isolated from patients, especially from the urinary tracts of the catheterised patients. The infections associated with biomaterials are crucial therapeutic obstacles, due to the bactericidal resistance of the biofilm. The aim of this study was to assess the susceptibility of P. mirabilis planktonic forms to ciprofloxacin and ceftazidime, the ability to form biofilm, and the impact of chosen sub-MIC concentrations of these antibiotics on biofilm at different stages of its formation. The research included 50 P. mirabilis strains isolated from wounds and the urinary tracts from patients of the University Hospital No. 1 in Bydgoszcz. The assessment of susceptibility to ciprofloxacin and ceftazidime was conducted using micromethods. The impact of sub-MIC concentrations of the chosen antibiotics on the biofilm was measured using the TTC method. The resistance to ciprofloxacin was confirmed for 20 strains (40.0%) while to ceftazidime for 32 (64.0%) of the tested P. mirabilis strains. All of the tested strains formed biofilm: 24.0% weakly, 26.0% moderately, and 50.0% strongly. It was determined that ciprofloxacin and ceftazidime caused eradication of the biofilm. Moreover, the connection between origin of the strains, biofilm maturity level, and resistance to antibiotics was proved.

Antimicrobial susceptibility and genetic similarity of ESBL-positive Klebsiella pneumoniae strains.

Introduction Twenty-eight isolates of extended-spectrum β-lactamase (ESBL)-positive Klebsiella pneumoniae were studied. Material and methods The strains were cultured from different clinical specimens obtained from children hospitalised at the University Hospital in Bydgoszcz. Seventeen strains were isolated from colonization and eleven from clinical infection. Isolation and identification of bacteria were performed using routine methods at the clinical microbiology laboratory. Production of ESBL was assessed using the double disk synergy test. The susceptibility to imipenem and tigecycline was tested by the Etest. The susceptibility to gentamicin and ciprofloxacin was tested by the agar dilution method. The genomic DNA was extracted from the strains separated by pulsed-field gel electrophoresis (PFGE) after digesting with XbaI endonuclease. Results Among analysed K. pneumoniae strains all were susceptible to imipenem, 21 (75.0%) were susceptible to tigecycline, 14 (50.0%) to gentamicin and 5 (17.9%) to ciprofloxacin. Molecular typing results revealed a great genetic diversity among K. pneumoniae isolates. All repeated PFGE patterns were detected in seven K. pneumoniae isolates. Among identical K. pneumoniae strains four susceptibility patterns were detected. Conclusions The results of the study suggest that establishing strains’ similarity in epidemiological investigations should be based on results obtained by several methods, and that each phenotyping method should be complemented with genetic research.