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Radu Agrosoaie, Adrian Streinu-Cercel, Doina Azoicai, Codrina Bejan, Olga Dorobat, Alexandru Mihai, Mona Popoiu and Alexandru Rafila

Abstract

Introduction: Invasive fungal infections have stood as an important research subject for the past 20 years, being considered as a crucial effect of advancing healthcare services. Low identification rates of invasive fungal infections in blood cultures and low sensibility of biomarkers determine empiric treatments which lead to a change in epidemiological data and antifungal susceptibility. The aim: The epidemiological evaluation of invasive fungal infections and the assessment of antifungal resistance related to this condition. Methods and material: An “antifungal stewardship” retrospective study was developed between January 2010 and April 2016. An epidemiological analysis was performed on 79 cases with proven invasive fungal infections in bloodstream, catheter, and cerebrospinal fluid. We considered: age, gender, HIV status, place of residence, and first option in medical practice of antifungal treatment. The laboratory analysis was performed by the Microbiology Laboratory at “Prof. Dr. Matei Bals” National Institute for Infectious Diseases, Bucharest. Minimum inhibitory concentrations (MIC’s) of 15 isolates were identified using colorimetric micro broth dilution panel YEASTONE ®YO10 and compared with susceptibilities obtained by VITEK2®C system. Candida parapsilosis ATCC 22019 was used as reference. Results: The incidence of invasive fungal infections was 3.7 on 1000 hospitalized patients. The age of the study population ranged between 12 and 83 years, and most were male (59%). The majority of subjects were from an urban area (84%), and 27% of them were HIV positive. The results obtained in VITEK2C® were similar with those from YEASTONE® YO10 for fluconazole, voriconazole, amphotericin B (100%), without any minor, major or very major errors. The fluconazole was the first option of treatment, followed by voriconazole, caspofungin, anidulafungin. In 37% of cases the first treatment option was replaced with a secondary antifungal therapy accordingly with antifungal breakpoints obtained by Vitek ®. Conclusions: No rates of resistance to fluconazole, amphotericin B, voriconazole were obtained. Fluconazole was the major first line antifungal therapy. Conclusions: No rates of resistance to fluconazole, amphotericin B, voriconazole were obtained. Fluconazole was the major first line antifungal therapy.

Open access

Dragos Florea, Steliana Huhulescu, Alexander Indra, Ioana Badicut, Alexandru Rafila, Dan Otelea and Gabriel Adrian Popescu

Abstract

In recent years Clostridium difficile infection (CDI) has represented a serious public health issue, mainly due to the global spread of the hypervirulent strain NAP1/027/BI. The purpose of the present study was to evaluate the utility of a PCR coupled with electrospray ionization mass spectrometry (ESI-MS) commercial assay for the detection of C. difficile virulence markers. Non-duplicative C. difficile isolates from patients with CDI diagnosed in a tertiary level hospital from Bucharest were tested for toxin A, toxin B, binary toxin genes and deletion in tcdC gene using PCR/capillary gel electrophoresis and PCR/ESI-MS. The study analysed 45 non-duplicative isolates, 33 strains (73.3%) belonging to ribotype 027. The concordance between PCR/capillary gel electrophoresis and PCR/ESI-MS was 100% for toxin A gene, 97.8% for toxin B gene, 91.1% for binary toxin subunit A gene and 95.6% for binary toxin subunit B gene. The general concordance for the complete panel of markers was 88.9% but was 100% for ribotype 027 isolates. PCR/ESI-MS might be a valid method for the detection of C. difficile virulence markers, including binary toxin.

Open access

Alexandru Rafila, Daniela Talapan, Olga Mihaela Dorobăţ, Gabriel Adrian Popescu, Daniela Piţigoi, Dragoş Florea and Florin Corneliu Buicu

Abstract

Introduction: Hospital-acquired infections caused by Enterobacteriaceae producing different types of carbapenem- hydrolizing enzymes are now commonly observed and represent a great limitation for antimicrobial therapy. The purpose of the study was to evaluate the emergence of carbapenem-resistant Enterobaceriaceae among the strains isolated from hospitalized patients to the National Institute of Infectious Diseases, Bucharest (NIID) and the identification of different types of carbapenemases, using phenotypic methods.

Materials and methods: Between January - June 2014, 587 strains of Klebsiella pneumoniae, Enterobacter species and E.coli were isolated from various clinical specimens. We were included all non-susceptible strains to carbapenems, according to EUCAST 2014 clinical breakpoints, as determined by using microdilution MicroScan Panels (Siemens Healthcare Diagnostics). The modified Hodge test (MHT) was performed as phenotypic confirmatory test for carbapenemase production according to CLSI guidelines and the combination disk test (KPC, MBL , OXA-48 Confirm kit, Rosco Diagnostica) according to EUCAST guidelines.

Results: A total of 45 non-repeat Enterobaceriaceae (32 strains Klebsiella pneumoniae, 5 strains E.coli, 8 strains Enterobacter spp) were identified as non-susceptibile to one or more carbapenems (93,33% ertapenem, 53,33% meropenem, 48,88% imipenem). Most strains were isolated from urine (75,55%). MHT was positive in 55,6% (25/45) of carbapenem-resistant strains; in 24 cases the carbapenem-hydrolizing enzyme was identified as: OXA-48-like (n=16), KPC (n=4), MBL (n=1), KPC + MBL (n=2) and MBL + OXA-48-like (n=1). All carbapenemase- positive strains were 100% resistant to 3rd and 4th generation cephalosporins, showing less resistance to tigecycline (12,5% resistant and 25% intermediate), colistin (37,5%) and fosfomycin (41,6%).

Conclusion: During 6 months period, there were isolated 7,66% (45/587) carbapenem-resistant Enterobacteriaceae (K. pneumoniae 21,47%, E. coli 1,23%). Twenty four strains were carbapenemase-producers. The most frequent carbapenemase isolated in our study was OXA-48-like.

Open access

Marius Eduard Caplan, Lorena Andreea Mateescu, Tatiana Vassu Dimov, Alexandru Rafila and Alina Maria Borcan

Abstract

Listeria monocytogenes has a ubiquitous distribution in nature and could contaminate food of animal origin, causing severe infections in humans. Till present, little is known about the antibiotic resistance profiles of these strains in Romania. The aim of this study was to determine the antibiotic susceptibility patterns of 37 L. monocytogenes strains isolated from animal derived foods and from clinical samples. Food samples were collected from meat and dairy products, between 2009 and 2013. Clinical samples were collected from patients with septicemia, meningitis/meningo-encephalitis, abortion cases and newborns, hospitalized during April 2010 - April 2013 in three medical institutions from Bucharest: Babes Hospital, Elias Hospital, National Institute of Infectious Diseases (INBI) Matei Bals. All tested isolates exhibited resistance to cephalosporins and nalidixic acid; one strain isolated from boiled shell snails was resistant to trimethoprim/sulfamethoxazole. The resistance to the first choice antibiotic ampicillin in L. monocytogenes strains isolated from severe infections is underlining the need of in vitro antibiotic susceptibility testing of each clinical isolate to establish the efficacy of different antibiotics, as well as of extended epidemiological studies to highlight the resistance profiles of L. monocytogenes strains circulating in our country.

Open access

Cristina Popescu, Gabriel Adrian Popescu, Olga Dorobăț, Alexandru Rafilă, Diana Tănase, Claudia Mikula, Gudrun Weissensteiner and Steliana Huhulescu

Abstract

We report first description of clinical cases of OXA-48 carbapenemase-producing Klebsiella pneumoniae originating from patients hospitalized in the most important Infectious Diseases Hospital from Romania, between December 2012 and March 2013. All strains were isolated from patients who were previously admitted in surgical wards. None of the patients had been admitted in a hospital outside of Romania.