Ewa Swoboda-Kopeć

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.

Significance of polymethylmethacrylate (PMMA) modification by zinc oxide nanoparticles for fungal biofilm formation

The objective of this study was to obtain a material composite with antifungal properties for dentures to be used as an alternative protocol in denture stomatitis treatment and prevention. Denture stomatitis is still a clinical problem in patients particularly vulnerable to this disease. Composites of PMMA and doped ZnO-NPs (weight concentrations, 2.5%, 5%, 7.5%) and PMMA with sprayed solvothermal and hydrothermal ZnO-NPs were tested. The following investigations of newly formed biomaterials were undertaken: influence on Candida albicans solution, biofilm staining, XTT analysis and a quantitative analysis of adhered C. albicans. These studies evidenced the antifungal activity of both nanocomposites PMMA-ZnO-NPs and the efficacy of sputtering of zinc oxide nanoparticles on the PMMA. The study of the biofilm deposition on the surface showed that antifungal properties increase with increasing concentration of ZnO-NPs. The XTT assay in conjunction with testing the turbidity of solutions may indicate the mechanism by which ZnO-NPs exert their effect on the increased induction of antioxidative stress in microorganism cells. The denture base made of the aforesaid materials may play a preventive role in patients susceptible to fungal infections. Based on the results obtained a modified treatment of stomatitis Type II (Newtons classification) complicated by fungal infection was proposed.

Candida nivariensis in comparison to different phenotypes of Candida glabrata

The purpose of the study was to establish the prevalence of new Candida glabrata complex species: Candida nivariensis and Candida bracarensis isolated from clinical material, evaluate their phenotypes and the prevalence of gene family encoding extracellular glycosylphosphatidylinositol‐linked aspartyl proteases, crucial for C. glabrata virulence. Study material included 224 C. glabrata clinical strains. Candida glabrata phenotypes were identified using CHROMagar Candida medium. Strains were analysed by using C. glabrata‐specific PCR for the internal transcribed spacer region to confirmed the identification. To identify C. nivariensis and C. bracarensis strains, the D1/D2 region of 26S rRNA was sequenced. The prevalence of YPS‐family proteases genes was detected using standard PCR method. Candida nivariensis amounted about 6% among the total number of C. glabrata strains. Candida nivariensis strains had a white phenotype on chromogenic agar media and assimilated two sugars – trehalose and glucose. Among the 13 C. nivariensis strains, 10 did not present any YPS‐family protease genes. Coexistence of all detected YPS‐family protease genes was specific for C. glabrata species. This study identified C. nivariensis strains; however, no C. bracarensis strains were identified. The white phenotype of C. nivariensis was confirmed. Most strains of the new species do not present any of the tested YPS genes.

Epidemiology and susceptibility to antifungal agents of fungi isolated from clinical specimens from patients hospitalized in the Department of General and Liver Surgery of the Medical University of Warsaw

The aim of this study was to analyze the type and antibiotic susceptibility of fungi isolated from clinical specimens obtained from patients hospitalized in the Department of General, Transplantation and Liver Surgery of the Medical University of Warsaw between 2000 to 2002. Among the 326 clinical samples found to be positive on mycological culture, 356 strains were cultured. The most common isolates were yeastlike fungi of the genus Candida 334 (93.8%), while others included 33 other types (6.2%). The most commonly isolated species were Candida albicans, 194 strains (54.5%); Candida glabrata, 68 (19.1%); Candida krusei, 20 (5.6%); Candida inconspicua, 20 (5.6%); Candida tropicalis, 17 (4.8%); and Candida parapsilosis, 6 (1.7%). Upon testing for susceptibility to antifungal agents, all strains were susceptible to amphotericin B, while 43.8% of strains showed intermediate susceptibility to fluconazole and 25.3%, to itraconazole. Control of fungal infections in transplant and in immunocompromised patients is hindered by the low percentage of strains susceptible to commonly used antifungal agents, particularly of the triazole group.

Molecular and Phenotypic Characteristics of Methicillin-resistant Staphylococcus aureus Strains Isolated From Hospitalized Patients in Transplantation Wards

OBJECTIVES Hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) frequently causes therapeutic problems and provides information about the epidemiological condition of the ward. MATERIALS AND METHODS HA-MRSA isolated from patients on transplantation wards in 1991, 1994, 1996, and from 2005 to 2007 were compared using molecular methods such as restriction fragment length polymorphism-pulse field gel electrophoresis, multilocus sequence typing (MLST), multiplex polymerase chain reaction (PCR) for detection type of staphylococcal chromosomal cassette mec, and PCR for detection. RESULTS The analysis covered HA-MRSA strains, each from a different patient. All organisms were typed using molecular methods. MLST results were compared with an international base. The examined strains belonged to five different worldwide known clonal complexes: CC8 (78%), CC5 (12%), CC1 (4%), CC30 (2%), and CC51 (4%). All could be recognized as representatives of a clonal complex CC8 clones: ST239-III (sequence type 239 and SCCmec type III named EMRSA-1, -4, -11, Brasilian, Hungarian) occurred with a frequency of 35.9%, ST254-IV (EMRSA-10, Hannover) occurred in 33.3%, ST247-I (EMRSA-5,-7, Iberian) occurred in 20.5%, ST241-III (Finland-UK) occurred in 5.15%, and ST8-IV (EMRSA-2,-6) occurred in 5.15%. CONCLUSION The predomination of different clones of HA-MRSA in the particular years was observed. In 1991, the EMRSA-10 (Hannover) clone predominated (53.3%). The Brasilian-Hungarian (EMRSA-1, -4, -11) clone predominated in 1994 (50%) as well as from 2005 to 2007 (41.3%), whereas in 1996 the Iberian clone was most frequent (53.9%).

Staphylococcus aureus nasal carriage in Ukraine: antibacterial resistance and virulence factor encoding genes

BackgroundThe number of studies regarding the incidence of multidrug resistant strains and distribution of genes encoding virulence factors, which have colonized the post-Soviet states, is considerably limited. The aim of the study was (1) to assess the Staphylococcus (S.) aureus nasal carriage rate, including Methicillin Resistant S. aureus (MRSA) strains in adult Ukrainian population, (2) to determine antibiotic resistant pattern and (3) the occurrence of Panton Valentine Leukocidine (PVL)-, Fibronectin-Binding Protein A (FnBPA)- and Exfoliative Toxin (ET)-encoding genes.MethodsNasal samples for S. aureus culture were obtained from 245 adults. The susceptibility pattern for several classes of antibiotics was determined by disk diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. The virulence factor encoding genes, mecA, lukS-lukF, eta, etb, etd, fnbA, were detected by Polymerase Chain Reaction (PCR).ResultsThe S. aureus nasal carriage rate was 40%. The prevalence of nasal MRSA carriage in adults was 3.7%. LukS-lukF genes were detected in over 58% of the strains. ET-encoding genes were detected in over 39% of the strains and the most prevalent was etd. The fnbA gene was detected in over 59% of the strains. All MRSA isolates tested were positive for the mecA gene. LukS-lukF genes and the etd gene were commonly co-present in MRSA, while lukS-lukF genes and the fnbA gene were commonly co-present in Methicillin Sensitive S. aureus (MSSA) isolates. No significant difference was detected between the occurrence of lukS-lukF genes (P > 0.05) and the etd gene (P > 0.05) when comparing MRSA and MSSA. The occurrence of the fnbA gene was significantly more frequent in MSSA strains (P < 0.05).ConclusionsIn Ukraine, S. aureus is a common cause of infection. The prevalence of S. aureus nasal carriage in our cohort of patients from Ukraine was 40.4%. We found that 9.1% of the strains were classified as MRSA and all MRSA isolates tested positive for the mecA gene. We also observed a high prevalence of PVL- and ET- encoding genes among S. aureus nasal carriage strains. A systematic surveillance system can help prevent transmission and spread of drug resistant toxin producing S. aureus strains.

Influence of Denture Plaque Biofilm on Oral Mucosal Membrane in Patients with Chronic Obstructive Pulmonary Disease

Patients with chronic obstructive pulmonary disease (COPD) have the lower airways colonized with pathogenic bacteria in a stable period of the disease and during exacerbations. The etiology of bacterial exacerbations of COPD depends on the underlying disease, the frequency of exacerbations and antibiotic therapy. Microorganisms can be aspirated off the denture plaque biofilm into the lower respiratory tract and could reduce the patients immunity and cause pneumonia. COPD patients, who are using acrylic dentures in oral cavity, are exposed to denture stomatitis and oral candidiasis. The aim of this study was to establish the composition of denture plaque biofilm and its impact on the oral mucosa in COPD patients. The study included patients in a stable phase of COPD using removable denture and the control group included healthy wearers appliances. Examinations concerned the oral mucosal membrane and the hygienic condition of prosthetic restorations. Microbiological examinations were performed by taking a direct swab from the surface of acrylic dentures. Seventeen bacterial and fungal strains were isolated from denture plaque of COPD patients, which could be a reservoir of pathogens in the upper and lower airways. The results showed a greater frequency of prosthetic stomatitis complicated by mucosal infections among COPD patients compared to healthy subjects.

Acinetobacter baumannii Multidrug-Resistant Strain Occurrence in Liver Recipients With Reference to Other High-Risk Groups

INTRODUCTION The increasing clinical significance of Acinetobacter baumannii species is due to its ability to survive in hospital environments, its species-specific multidrug resistance, and its ability to instantly develop various drug-resistance mechanisms through antibiotic pressure. MATERIALS AND METHODS We identified 16 A baumannii strains isolated from patients presenting postoperative infections in 2010. A baumannii isolates were obtained from clinical specimens by standard microbiologic methods. As previously described, we performed polymerase chain reaction (PCR) analysis for carbapenemase-encoding genes (VIM, IMP, SPM, OXA23, OXA24, OXA51, OXA58) in Acinetobacter spp. RESULTS The double-disk synergy test phenotypic method did not detect any A baumannii strains producing metallo-beta-lactamaus cultured from swabs from all the patient groups. No products of PCR amplification with specific starters for VIM, IMP, and SPM (Sao Paulo metallo-β-lactamase) genes were found. All analyzed strains were colistin-sensitive. Among five strains from liver recipients, one was imipenem- and meropenem-resistant. Four among six strains isolated from cancer patients were resistant to imipenem and/or meropenem; 1/5 were imipenem-and meropenem-resistant; 1, meropenem-resistant and imipenem-sensitive; 1, meropenem- and imipenem-resistant; and 1 with intermediate resistance to both meropenem and imipenem among swabs cultured from patients with postoperative complication after bone fracture. Fifteen among 16 analyzed A baumannii strains had an OXA51 gene. Two among five A baumannii strains isolated in liver recipients had only an OXA51 gene; one, OXA51 and OXA24 genes; one, OXA51 and OXA23 genes.

Diagnosis of Invasive Pulmonary Aspergillosis

Culturing strains from clinical samples is the main method to diagnose invasive pulmonary aspergillosis. Detecting the galactomannan antigen in serum samples is an auxiliary examination. The goal of this study was to determine the frequency with which Aspergillus fumigatus was cultured in clinical samples taken from patients hospitalized in the the Infant Jesus Teaching Hospital in Warsaw, Poland, in the period of 2013-2014. Specimens from the respiratory tract and blood were cultured for mycological and serological assessments. Strain isolation was performed in chloramphenicol Sabouraud agar. Species identification was based on morphological traits in macro-cultures and on microscopic examination. The galactomannan antigen was detected by ELISA method. Out of 2000 clinical samples with positive mycological results, 200 were obtained from the respiratory tract. A. fumigatus was cultured in 13 cases from the respiratory group. Ten cases were cultured out of tracheal aspirates and three from bronchoalveolar lavage fluid. The galactomannan antigen was detected in a serum sample from only one out of the 13 patients with cultures positive for A. fumigatus. It also was detected in serum samples of three other patients in whom A. fumigatus culture yielded a negative result. We conclude that culture-confirmed invasive pulmonary aspergillosis represents a scarce finding. A. fumigatus cultured from clinical samples may not always be confirmed by ELISA assay and vice versa a positive ELISA result does not attest the successful culture.

Aspergillus galactomannan detection in comparison to a real-time PCR assay in serum samples from a high-risk group of patients.

Invasive aspergillosis (IA) is a severe infection with a 70% mortality rate. Aspergillus fumigatus is responsible for over 90% of those infections. The diagnosis of invasive aspergillosis is based on clinical sample culture and detection of fungal hyphae in histopathological examination. Additional tests may include the detection of the galactomannan antigen and of fungal genetic material in serum and bronchoalveolar washings. The present study was to assess the use of these two rapid tests in the diagnosis of invasive aspergillosis: serological one – to detect the galactomannan antigen (ELISA assay), and real-time PCR, and to establish a possible correlation between these two methods.