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.

Trends in antifungal susceptibility of Candida species--one year observation.

In the past years opportunistic fungal infections have seriously increased, mainly in immunocompromised patients. The aim of the study was to determine the prevalence of yeast-like fungi in invasive candidiasis and to estimate its susceptibility to chosen antifungal agents. One hundred and sixty strains of yeast-like fungi were cultured from various clinical material: samples from lower respiratory tract, blood, the peritoneal cavity and others. The susceptibility tests were established according to the quantitative E-test method. The Candida genus represented the main etiological factor of invasive candidiasis. The predominant species were: C. glabrata (71/160), C. albicans (34/160), C. krusei (17/160), C. tropicalis (14/160). All tested strains were the most resistant to itraconazole. Candida glabrata presented the 100% susceptibility to amphotericin B and caspofungin and was the least susceptible to itraconazole, posaconazole and voriconazole. Candida albicans was the most susceptible species to all antymicotics.

Polymerase chain reaction melting profiles of Candida glabrata clinical isolates in solid organ transplant recipients in comparison to the other group of surgical patients.

The aim of the retrospective study were to estimate the prevalence of Candida glabrata in liver and kidney transplant recipients compared to patients with short bowel syndrome receiving chronic total parenteral nutrition and relevance of the polymerase chain reaction melting profile (PCR MP) method for Candida glabrata strains differentiation. C. glabrata clinical strains isolated from patients were identified by using standard mycological procedures. The analysis of genetic relatedness of the isolated strains was conducted using the PCR MP method. The prevalence of C. glabrata comprised 29% of all episodes of fungal colonization and infection in solid organ transplant recipients, and 54% of those in hospitalized patients receiving long-term total parenteral nutrition. Among 78 isolates obtained from 55 solid organ transplant recipients and 2 organ donors, 44 different C. glabrata PCR MP fingerprints were observed. Forty-seven organ recipients and one organ donor carried unique C. glabrata strains. Among 37 isolates obtained from 31 patients receiving long-term TPN, 8 different PCR MP profiles of C. glabrata strains were observed. Two patients carried unique C. glabrata strains. Most of the C. glabrata colonization and infections in solid-organ transplant recipients were caused by endogenic strains. Most of the C. glabrata colonization and infections in hospitalized patients receiving long-term total parenteral nutrition could result by patient-to-patient transmission. The results showed that the PCR MP technique is a good discriminatory method for genotyping for C. glabrata strains.

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-ß-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.

The probability of the Acinetobacter baumannii strain clonal spreading in donor-recipient systems, as confirmed by the molecular analysis of randomly amplified polymorphic DNA.

Acinetobacter baumannii is an important pathogen widely distributed in the hospital environment and responsible for a variety of nosocomial infections. This micro-organism especially affects patients with impaired host defenses in the intensive care unit. It has been implicated in severe nosocomial infections including bloodstream infections, pneumonia, and meningitides. Those infections are often outbreaks caused by a single clone spreading. The aim of our study was an epidemiological analysis of Acinetobacter baumannii strains isolated from hospitalized liver/kidney transplant donors and recipients. The analyzed material for epidemiological test included 13 A. baumannii strains isolated in 2010 from eight liver/kidney donors and 5 organ recipients. The epidemiological analysis of the isolates was performed by the use of the random amplified polymorphic DNA (RAPD)-polymerase chain reaction method to determine their genetic relatedness. We isolated 9 A. baumannii strains from 8 organ donors. Among this group of isolates, four strains showed the same fingerprints that were classified as one RAPD type 1. The remaining donor isolates revealed differentiated patterns. All strains isolated from recipients formed distinct RAPD types, one of which was identical to the group of four donor strains (RAPD type 1). The clonal spreading of A. baumannii strains was not observed among recipients but we noted a single case of probable transmission of the pathogen from the donor to the recipient.

Differences in proteolytic activity and gene profiles of fungal strains isolated from the total parenteral nutrition patients.

Fungal infections constitute a serious clinical problem in the group of patients receiving total parenteral nutrition. The majority of species isolated from infections of the total parenteral nutrition patients belong to Candida genus. The most important factors of Candida spp. virulence are the phenomenon of "phenotypic switching," adhesins, dimorphism of fungal cells and the secretion of hydrolytic enzymes such as proteinases and lipases, including aspartyl proteinases. We determined the proteolytic activity of yeast-like fungal strains cultured from the clinical materials of patients receiving total parenteral nutrition and detected genes encoding aspartyl proteinases in predominant species Candida glabrata--YPS2, YPS4, and YPS6, and Candida albicans--SAP1-3, SAP4, SAP5, and SAP6. C. albicans released proteinases on the various activity levels. All C. glabrata strains obtained from the clinical materials of examined and control groups exhibited secretion of the proteinases. All 13 isolates of C. albicans possessed genes SAP1-3. Gene SAP4 was detected in genome of 11 C. albicans strains, SAP5 in 6, and SAP6 in 11. Twenty-six among 31 of C. glabrata isolates contained YPS2 gene, 21 the YPS4 gene, and 28 the YPS6 gene. We observed that clinical isolates of C. albicans and C. glabrata differed in SAPs and YPSs gene profiles, respectively, and displayed differentiated proteolytic activity. We suppose that different sets of aspartyl proteinases genes as well as various proteinase-activity levels would have the influence on strains virulence.

Algorithm of clinical protocol lowering the risk of systemic Mycosis infections in allografts recipients.

The aim of the study was to describe a diagnostic protocol to lower the risk of a mycotic invasive infection among allotransplant recipients and to suggest the use of preoperative prophylaxis and/or empiric therapy. We chose a group of 268 allograft recipients with transient or constant yeast colonization or confirmed yeast infection. Among 7744 clinical samples, 475 were positive for fungi. We used conventional fungal laboratory diagnosis, enzymatic activity tests, serologic tests, molecular diagnosis of samples from sterile body sites, and histopathologic examinations. The following clinical samples were examined: blood samples; swabs from mouth lesions, throat, and rectum; and sputum, urine, and fecal samples from kidney transplant recipients and simultaneous pancreas-kidney transplantation recipients who are highly predisposed to mycotic infections. We established microbiologic criteria of a systemic mycosis and principles to distinguish colonization from infection.

Trichosporon asahii as a prospective pathogen in solid organ transplant recipients.

Trichosporon spp. is not an important factor of mycotic infections in immunocompetent patients. It may be a cause of invasive mycoses with a high mortality rate in patients undergoing solid organ transplantation. We have analysed the antifungal agents' susceptibility of Trichosporon asahii and its frequency of occurrence as a prospective etiological agent of infections in liver, kidney and simultaneous pancreas-kidney transplant recipients. Clinical specimens (urine, blood, peritoneal fluid and swabs) were obtained from patients hospitalised in the Institute of Transplantation Medicine, Department of General and Transplantation Surgery, Medical University of Warsaw in 2005 and 2006. Microbiological tests were performed in Mycological Laboratory, Department of Microbiology, Medical University of Warsaw. A total of 475 strains of yeast-like fungi were isolated from clinical specimens taken from 263 liver, kidney and simultaneous pancreas-kidney transplant recipients and from 26 organ donors. Trichosporon asahii was found in 26 clinical samples taken from 18 patients and one organ donor. Positive cultures were obtained from 22 urine samples, one stoma fluid, one wound swab, one tracheal aspirate and one ejaculate. Isolates of Trichosporon asahii were found in 6% of total positive mycological cultures in the solid organ transplant recipients. Among cultured strains, 11 isolates were resistant to fluconazole, four to itraconazole and three of them demonstrated resistance to amphotericin B.