[show abstract][hide abstract] ABSTRACT: Infections caused by Malassezia yeasts are most likely underdiagnosed, because fatty acid supplementation is needed for growth. Rapid identification of Malassezia species is essential for appropriate treatment of Malassezia related skin-infections, fungemia and nosocomial outbreaks in neonates, children and adults and can be live saving for those patients. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was reported as a rapid and reliable diagnostic tool to identify clinically important yeasts, but so far no data have been reported on identification of Malassezia isolates with this technique.
To create an extensive database of main mass spectra (MSPs) that will allow quick identification of Malassezia species by MALDI-TOF MS.
An in-house library of 113 MSPs was created from 48 reference strains from the CBS-KNAW yeast collection. The in-house library was challenged with two test sets of Malassezia strains, namely 165 reference strains from the CBS collection and 338 isolates collected in Greece, Italy, Sweden, and Thailand.
MALDI-TOF MS allowed correct identification of all 14 Malassezia spp. MALDI-TOF MS results were concordant with those of sequence analyses of the internal transcribed spacers (ITS1/ITS2) and the D1/D2 domains of the large subunit (LSU) of the ribosomal DNA (rDNA).
Implementation of the MALDI-TOF MS system as a routine identification tool will contribute to correct identification of Malassezia yeasts with a minimal effort and in a short turnaround time which is specifically important for the rapid identification of Malassezia in skin diseases and nosocomial outbreaks. This article is protected by copyright. All rights reserved.
British Journal of Dermatology 10/2013; · 3.76 Impact Factor
[show abstract][hide abstract] ABSTRACT: This study furnishes deeper insights to previous works on anidulafungin, demonstrating the potent activity against Candida strains planktonic cells and biofilms. Candida sp., associated with many biomaterial-related infections, give rise to infective pathologies typically associated with biofilm formation. We recently determined the in vitro antifungal activities of echinocandin anidulafungin in association with some antifungal drugs against some Candida strains in their planktonic states. A total of 11 Candida strains biofilms were tested in this study: six Candida albicans, three C. parapsilosis and two C. tropicalis. All yeast isolates and ATCC strains were stored at -20 °C in glycerol stocks and were subcultured on antimicrobial agent-free Sabouraud dextrose agar plates. MIC endpoints were determined colorimetrically by using the indicator 2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) with menadione as electron-coupling agent. The activity of anidulafungin was assessed using in vitro microbiological model relevant for clinical practice. Anidulafungin showed a strong activity in vitro against both planktonic and biofilms cells, and our study confirms that high anidulafungin concentrations might establish paradoxical growth effect in C. albicans and C. tropicalis biofilms.The Journal of Antibiotics advance online publication, 11 September 2013; doi:10.1038/ja.2013.83.
The Journal of Antibiotics 09/2013; · 2.19 Impact Factor
[show abstract][hide abstract] ABSTRACT: Legionella spp. is considered an emerging microorganism involved in aquatic environments contamination and cause of Legionnaires' disease. The aims of the study are to evaluate the level of contamination of Legionella spp. in the water system of the largest Hospital of Apulia region during a 4-year surveillance and to establish, by molecular method, the presence of a predominant genotype of L. pn. sg 1. The results showed that Legionella spp. was present in 36% of water samples with Legionella pneumophila serogroup 1 (L. pn. sg 1) the most prevalent species and serogroup and the wards most contaminated are the high risk units. In addition, despite four main clones of L. pn. sg 1 were identified, a predominant genotype existed. In conclusion the study demonstrates the necessity for periodic evaluation on hospitals water system to assess the potential contamination of Legionella spp., performing decontamination in the presence of bacterial contamination, even low, in particular in high risk wards. Moreover, the switching of the disinfection methods may be suggested in order to prevent resistance phenomenon by some L. pn. sg 1 clones.
Annali di igiene: medicina preventiva e di comunità 09/2013; 25(5):435-41.
[show abstract][hide abstract] ABSTRACT: BACKGROUND: The training of health workers is a key issue for the prevention of healthcare associated infections. OBJECTIVES: To evaluate knowledge of nursing and medical students concerning the prevention of healthcare associated infections. DESIGN: A cross-sectional study. SETTING: University hospitals in nine Italian cities. PARTICIPANTS: One thousand four hundred sixty one healthcare students (607 medical students and 854 nursing students). METHODS: The study was performed using a questionnaire investigating 3 areas, each having different possible points: standard precautions=12; hand hygiene=8; healthcare associated infections=5, for an overall perfect score of 25. Scores that met a cutoff ≥17.5 were considered to be indicative of an acceptable level of knowledge. Factors associated with an acceptable level of knowledge were analyzed using a logistic regression model. RESULTS: Mean overall score (±SD) was 18.1±3.2. Nursing students (18.6±2.9) obtained a higher overall score than medical students (17.4±3.5) (p<0.001). Weighed scores (±SD) by area were: 10.3 (±2.0) for standard precautions, 5.0 (±1.3) for hand hygiene and 2.8 (±1.1) for healthcare associated infections. Knowledge level concerning the three areas was different between medical and nursing students (p<0.001). The probability of finding acceptable knowledge was smaller for medical students (OR: 0.54 p<0.0001) and for students aged ≥24years (OR: 0.39 p<0.0001). CONCLUSION: The overall score showed an acceptable level of knowledge for the whole sample; but, considering separately the two curricula, only nursing students reached the minimum acceptable score. It seems important to investigate what is working better in nursing than in medical education in order to implement and validate new teaching approaches.
[show abstract][hide abstract] ABSTRACT: PURPOSE: The aim of this study was to assess the epidemiology of candidemia and antifungal susceptibility profiles of Candida isolates in Italy through a prospective surveillance study and to evaluate changes compared to a previous survey performed in one Italian region (Lombardy) in 1997-1999. METHODS: A prospective laboratory-based surveillance of candidemia was performed in Italy from January to December 2009. For each case a questionnaire was filled in, and the first isolate was collected and tested for in vitro antifungal susceptibility. RESULTS: During our 12-month survey, 467 episodes of candidemia were reported from 34 centres (30 located in Lombardy) and 464 isolates collected. Candida albicans was the predominant species (overall incidence 50.4 %), but the proportion varied considerably from 52.1 % in Lombardy hospitals to 45.2 % hospitals located outside this region. The second most frequent species was C. glabrata in Lombardy and C. parapsilosis in other regions. Comparison of the 1997-1999 and 2009 data on episodes of candidemia in Lombardy revealed a threefold increase in incidence (from 0.38 to 1.19 per 1,000 admissions), aging of infected patients, decline in crude mortality (from 35 to 27.1 %) and an increased proportion of C. glabrata etiology (from 12.8 to 20.3 %). Susceptibility testing confirmed the broad activity of amphotericin B and echinocandins. Decreased susceptibility to fluconazole was found in 24.9 % of the tested isolates. CONCLUSIONS: The results of this latest survey confirm the high rate of candidemia in Italy and show changes in some of the epidemiological tracts, such as aging of infected patients, increased proportion of C. glabrata infections, increased diagnosis in medical wards, and improvement in patients' survival.
[show abstract][hide abstract] ABSTRACT: BACKGROUND: Implementing infection control measures in light of healthcare-associated infection (HAI) surveillance data can prevent HAIs. Surveillance has been associated with a reduction of HAI in intensive care unit (ICU) patients, though the reasons for this improvement remain unclear. AIM: To evaluate changes in healthcare-associated infection (HAI) rates during three surveys of the Italian Nosocomial Infections Surveillance in Intensive Care Units (ICUs) network (SPIN-UTI) six-year project and to explore sources of variation of indicators of HAI in the 65 participating ICUs. METHODS: The SPIN-UTI network adopted the European protocols for patient-based HAI surveillance. Cumulative incidence, incidence density, infection rates adjusted for device-days, and device utilization ratios were calculated for each survey and compared. To identify risk factors multiple logistic regression analyses were performed. Crude excess mortality was computed as the difference between the crude overall case-fatality rate of patients with and without HAI. FINDINGS: The risk of ICU-acquired infections increased in the third survey compared with previous (relative risk: 1.215; 95% confidence interval: 1.059-1.394). Among risk factors, the number of hospitalized patients requiring ICU admission and the Simplified Acute Physiology Score II increased from 73.7% to 78.1% and from 37.9% to 40.8% respectively. Although mortality rates remained unchanged, HAIs trebled the risk of death. Acinetobacter baumannii was the most frequently reported micro-organism in the third survey (16.9%), whereas in the previous surveys it ranked third (7.6%) and second (14.3%). CONCLUSIONS: The study highlighted the increased risk of HAI, at least partially explained by the greater severity and number of hospitalized patients requiring ICU admission. Furthermore, the management of intubation procedures and of ventilated patients was identified as a potential target for infection control interventions to decrease the growing risk of HAI in ICUs.
The Journal of hospital infection 03/2013; · 3.01 Impact Factor
[show abstract][hide abstract] ABSTRACT: PURPOSE: The aims of this study are to evaluate the epidemiology of invasive fungal infections (IFIs) in patients admitted to an intensive care unit (ICU) in Southern Italy and the in vitro antifungal susceptibility of isolates. METHODS: A surveillance program was implemented in 18 ICUs. IFI cases were recorded using a standardized form. RESULTS: A total of 105 episodes of IFIs occurred in 5,561 patients during the 18-month study. The main infections were caused by yeasts, more than filamentous fungi (overall incidence of 16.5 cases per 1,000 admissions and 2.3 cases per 1,000 admissions, respectively). The overall crude mortality rate was high (42.8 %), particularly for mold infections (61.5 %). All yeast infections were Candida bloodstream infections. Over half (59.8 %) were caused by Candida non-albicans, with C. parapsilosis being the most common (61.8 %). In the multivariate model, trauma admission diagnosis, prolonged stay in the ICU, and parenteral nutrition were independently associated with candidemia due to C. parapsilosis [odds ratio (OR) 3.5, (1.8-5.2); OR 3.5, (1.02-3.5); OR 3.6, (1.28-6.99), respectively]. Among mold infections, 12 patients suffered from invasive pulmonary aspergillosis, with Aspergillus fumigatus as the predominant pathogen (41.7 %). One case of brain scedosporiosis was identified. Overall, azoles and echinocandins resistance was uncommon. CONCLUSIONS: Candida non-albicans species are the most frequent cause of candidemia in ICU patients. Mold infections are associated with a high mortality rate. This study confirms the importance of the epidemiological surveillance on IFIs in the ICU setting for documenting species distribution and antimicrobial susceptibility patterns to guide therapeutic choices.
[show abstract][hide abstract] ABSTRACT: To evaluate microbial contamination in hospitals environments, several methods are available, each one having its limitations. Therefore, the choice of system to use is open. This study compares the ability of a dusting cloth pad (DC pad) with 2 other methods (Rodac contact plate and air sampling) to detect contamination because of filamentous fungi in operating rooms, performing 110 sampling campaigns in hospitals of 3 Italian cities. Overall, 96% of the DC pad samples were positive compared with 51% of Rodac plates (P < .0001) and 35% of air samples (P < .0001). Authors conclude that the DC pad improves the ability to detect an environmental contamination of filamentous fungi.
American journal of infection control 02/2013; · 3.01 Impact Factor
[show abstract][hide abstract] ABSTRACT: Until recently, Cryptococcus gattii infections occurred mainly in tropical and subtropical climate zones. However, during the past decade, C. gattii infections in humans and animals in Europe have increased. To determine whether the infections in Europe were acquired from an autochthonous source or associated with travel, we used multilocus sequence typing to compare 100 isolates from Europe (57 from 40 human patients, 22 from the environment, and 21 from animals) with 191 isolates from around the world. Of the 57 human patient isolates, 47 (83%) were obtained since 1995. Among the 40 patients, 24 (60%) probably acquired the C. gattii infection outside Europe; the remaining 16 (40%) probably acquired the infection within Europe. Human patient isolates from Mediterranean Europe clustered into a distinct genotype with animal and environmental isolates. These results indicate that reactivation of dormant C. gattii infections can occur many years after the infectious agent was acquired elsewhere.
[show abstract][hide abstract] ABSTRACT: A number of factors, for example water temperature, can encourage the growth of microorganisms such as Legionella spp in spa facilities. Individuals who attend this type of facility are often subjects at risk for infection who are undergoing inhalation therapy and hot tub treatments. A very accurate management of these facilities is therefore required to avoid infection by Legionella spp. The purpose of this study was to verify the current Italian national and Apulia regional legislation regarding the control of contamination by Legionella spp. in spa facilities.
[show abstract][hide abstract] ABSTRACT: Since air can play a central role as a reservoir for microorganisms, in controlled environments such as operating theatres regular microbial monitoring is useful to measure air quality and identify critical situations. The aim of this study is to assess microbial contamination levels in operating theatres using both an active and a passive sampling method and then to assess if there is a correlation between the results of the two different sampling methods.
The study was performed in 32 turbulent air flow operating theatres of a University Hospital in Southern Italy. Active sampling was carried out using the Surface Air System and passive sampling with settle plates, in accordance with ISO 14698. The Total Viable Count (TVC) was evaluated at rest (in the morning before the beginning of surgical activity) and in operational (during surgery).
The mean TVC at rest was 12.4 CFU/m3 and 722.5 CFU/m2/h for active and passive samplings respectively. The mean in operational TVC was 93.8 CFU/m3 (SD = 52.69; range = 22-256) and 10496.5 CFU/m2/h (SD = 7460.5; range = 1415.5-25479.7) for active and passive samplings respectively. Statistical analysis confirmed that the two methods correlate in a comparable way with the quality of air.
It is possible to conclude that both methods can be used for general monitoring of air contamination, such as routine surveillance programs. However, the choice must be made between one or the other to obtain specific information.
BMC Public Health 08/2012; 12:594. · 2.08 Impact Factor
[show abstract][hide abstract] ABSTRACT: Candidemia is a major infectious complication in neonatal patients. The isolation of yeasts from blood is still the "gold standard" for its diagnosis, but other laboratory markers (i.e., circulating antigens) have been studied with varying specificities and sensitivities. The aim of this study was to evaluate the role of procalcitonin for the diagnosis of candidemia in neonatal patients at high risk. To verify if the use of different commercial methods can highlight dissimilar results of sensitivity and/or specificity, the determination of procalcitonin serum levels was estimated by two systems. Overall, 90 patients from a Neonatal Intensive Care Units were enrolled, of whom six developed Candida bloodstream infection. Four of six infants with candidemia had slight increase of procalcitonin values (0.5-1 ng/mL). Only one baby showed very high levels but he had fungal and bacterial sepsis at the same time, while no elevation was observed in the sixth patient. No statistically significant difference was observed between two different methods at the time of monitoring (p > 0.643). Both methods showed a sensitivity of 83.3 % at diagnosis, while the specificity was 73.8 and 63.1 % by methods A and B, respectively. In the light of the low sensibility and specificity of this assay, we can assume that the determination of procalcitonin would not seem to play a significant role in the diagnosis of fungal infection in neonatal patients.
[show abstract][hide abstract] ABSTRACT: Background/purposeAccording to WHO recommendations, 12-year-olds are considered an important target group for evaluating the level of dental caries among children with permanent teeth, and are often chosen for international comparisons. The aim of the present study was to evaluate the current oral health status of 12-year-old children in southeast Italy, stratified by gender and residential area.Materials and methodsThe survey was conducted on 431 children enrolled by multistage cluster sampling. A dental caries experience index (decayed, missing, and filled permanent teeth; DMFT) was recorded at schools by a team of examiners trained at the start of the study. Statistical analyses by Chi-square, Fisher's exact, and Wilcoxon tests were performed using SAS version 9.1 software for PCs. We applied the Zero-Inflated Negative Binomial regression model in the STATA package.ResultsCaries prevalence was recorded in 38.3% of the sample. Estimated means and 95% confidence intervals of the DMFT index by gender were: 1.15 (0.91–1.39) for males, 1.26 (1.02–1.5) for females, and 1.21 (1.03–1.39) for the total sample. The D component of the index was dominant. The mean number of caries found in southeast Italy was significantly higher than the national mean (t = 3.125, P = 0.002), but significantly lower than the mean for south Italy (t = −2.125, P = 0.03). Results of the regression model showed that only the mother and father's nationality and educational level contributed to the DMFT.Conclusions
The oral health situation of 12-year-old children from southeast Italy seems to be in line with that in other Western European countries.
[show abstract][hide abstract] ABSTRACT: The microdilution antifungal method (CLSI BMD, M27-A3) was used for testing the antifungal susceptibility of Malassezia species. However, optimal broth media that allow sufficient growth of M. pachydermatitis and produce reliable and reproducible MICs using the CLSI BMD protocol are yet to be established. In this study, the susceptibility of M. pachydermatis isolates to ketoconazole (KTZ), itraconazole (ITZ) and fluconazole (FLZ) was evaluated in vitro by the CLSI BMD test using Christensen's urea broth (CUB) and mRPMI 1640 containing lipid supplementation, Sabouraud dextrose broth with 1% tween 80 (SDB), and Dixon broth (DXB). A FLZ-resistant M. pachydermatis was generated in vitro and tested under the same conditions. A good growth of M. pachydermatis incubated for 48 and 72 h, respectively, was observed in CUB, SDB and DXB and not in mRPMI 1640 (p<0.001). No statistically significant differences were detected between the MIC values registered after 48 h and 72 h incubation. ITZ displayed lower MIC values than KTZ and FLZ regardless of the media employed. A large number of FLZ-resistant Malassezia strains (86.6%) was observed using DXB. A MIC>64 mg/L was observed only when the FLZ-resistant M. pachydermatis isolate was tested in SDB. Based on the results obtained herein, culture in SDB, stock inoculum suspensions of 1-5 × 10(6)CFU/ml, and an incubation time of 48 h are proposed as optimal conditions for the evaluation of the in vitro antifungal susceptibility of M. pachydermatis using a modified CLSI BMD protocol.
[show abstract][hide abstract] ABSTRACT: In this paper the authors investigated a synergistic antimycotic effect between four antifungal drugs Amphotericin B, Fluconazole, Tioconazole, and Flucytosine individually combined with Anidulafungin compound. This latter is considered a drug of choice in the treatment of fungal infections; it has good activity both in vitro and in vivo against yeasts and moulds, as Candida and Aspergillus. The goal of this study was to evaluate the in vitro interaction of Anidulafungin in the synergic combinations with previous reported drugs against 12 Candida strains according to CLSI M27-A3 protocol. A synergistic interaction was observed against the most antifungal strains; in particular an increasing of the antimycotic efficacy was obtained from the association between Anidulafungin and Amphotericin B or Fluconazole (Mixture 4:6). In contrast the association Tioconazole/Anidulafungin was less effective on fungal species growth. The antimycotics MIC reduction values were more evident against some strains as C. glabrata, C. krusei, C. tropicalis and C. parapsilosis.
[show abstract][hide abstract] ABSTRACT: INTRODUCTION: Candida prophylaxis in ICU is still a matter of debate. Oral chemoprophylaxis has been advocated to reduce the incidence of Candida colonisation and infection. METHODS: We performed a randomised trial studying a single drug (nystatin) versus control in surgical/trauma ICU patients. Multiple-site testing for fungi was performed in each patient on ICU admission (T0) and subsequently every 3 days (T3, T6, T9, and so forth). The primary evaluation criterion was the time course of the corrected colonisation index. RESULTS: Ninety-nine patients were enrolled. At admission, 69 patients exhibited Candida colonisation: the most frequently colonised body sites were the stomach and the pharynx. The most frequent isolated species was Candida albicans. The corrected colonisation index was similar in the two groups at T0 (P = 0.36), while a significant statistical difference was observed between the treatment and control groups at T6 (median 0.14 and 0.33, respectively; P = 0.0016), at T9 (median 0.00 and 0.28, respectively; P = 0.0001), at T12 (median 0.00 and 0.41, respectively; P = 0.0008), and at T15 (median 0.00 and 0.42, respectively; P <0.0003). The same results were obtained in the subgroup of patients already colonised at ICU admission. CONCLUSION: This trial shows that nystatin prophylaxis significantly reduces fungal colonisation in surgical/trauma ICU patients, even if already colonised. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01495039.
Critical care (London, England) 04/2012; 16(2):R57. · 4.72 Impact Factor
[show abstract][hide abstract] ABSTRACT: Dermatitis caused by Malassezia spp., one of most common skin disease in dogs, requires prolonged therapy and/or high doses of antifungal agents. In the present study, the antifungal susceptibility of M. pachydermatis to ketoconazole (KTZ), fluconazole (FLZ), itraconazole (ITZ), posaconazole (POS) and voriconazole (VOR) was evaluated in vitro using both CLSI reference broth microdilution (CLSI BMD) and E-test. A total of 62 M. pachydermatis strains from dogs with and without skin lesions were tested. M. pachydermatis strains were susceptible to ITZ, KTZ and POS using both test methods, with the highest MIC found in tests of FLZ. Essential agreement between the two methods ranged from 87.1% (VOR) to 91.9% (ITZ), and categorical agreement from 74.2% (FLZ) to 96.8% (ITZ). Minor error discrepancies were observed between the two methods, with major discrepancies observed for KTZ. A higher MIC(50) value for FLZ was noted with M. pachydermatis genotype B. The MICs(50) of M. pachydermatis genotype B for KTZ, VOR and POS were higher in isolates from dogs with skin lesions than those in isolates from animals without skin lesions. The results suggest a link between genotypes of M. pachydermatis and in vitro drug susceptibility. The categorical agreement for both E-test and CLSI BMD methods found in this investigation confirms the E-test as a reliable diagnostic method for routine use in clinical mycology laboratories.
Medical mycology: official publication of the International Society for Human and Animal Mycology 04/2012; 50(8):795-801. · 2.13 Impact Factor