To document reported Intubator Associated Pneumonia (IAP) prevention practices in Intensive Care Units (ICUs) and attitudes towards the implementation of a measurement system.

Rhino-cerebral zygomycosis (RCZ) is an acute rapidly progressive fungal infection usually occurring in patients with diabetes mellitus and ketoacidosis. Patients typically complain of pain located in the facial, nasal or orbital regions, followed by sudden blindness and cranial nerve palsy. Early diagnosis, correction of risk factors, prompt surgical removal and aggressive antifungal therapy are warranted as life-saving treatments. The following report describes a case of a lethal RCZ which occurred in an apparently healthy woman with latent non-decompensated diabetes mellitus and a fetal-type posterior (FTP) circle of Willis.

Legionella quantification in environmental samples is overestimated by qPCR. Combination with a viable dye, such as Propidium monoazide (PMA), could make qPCR (named then vPCR) very reliable. In this multicentre study 717 artificial water samples, spiked with fixed concentrations of Legionella and interfering bacterial flora, were analysed by qPCR, vPCR and culture and data were compared by statistical analysis. A heat-treatment at 55 °C for 10 minutes was also performed to obtain viable and not-viable bacteria. When data of vPCR were compared with those of culture and qPCR, statistical analysis showed significant differences (P < 0.001). However, although the heat-treatment caused an abatement of CFU/mL ≤1 to 1 log10 unit, the comparison between untreated and heat-treated samples analysed by vPCR highlighted non-significant differences (P > 0.05). Overall this study provided a good experimental reproducibility of vPCR but also highlighted limits of PMA in the discriminating capability of dead and live bacteria, making vPCR not completely reliable.

INTRODUCTION: Among the health professions with a long period of training, the students of the Nursing Bachelor's Degree are the most exposed to biological risk resulting from accidents, in particular with needles and cutting edges. The aim of the study was to estimate the frequency and the circumstances for the occurrence of needle stick injuries, as a knowledge base for targeted prevention interventions. METHODS: The study was carried out between May and July 2017 in 11 Universities in Italy and 1 in Albania (associated with the "Tor Vergata" University of Rome). An anonymous semi-structured questionnaire was proposed to 1st (second semester), 2nd and 3rd year students of Nursing Bachelor's Degree. RESULTS: A total of 2742 questionnaires were collected. The average age of participants was 22.9 years (median 22, range 19-60 years), 73% of whom were females. A total of 381 injuries were reported. Three hundred and sixteen students (11.8%) underwent at least 1 injury (12.7% among females, 9.7% among males); 41 students declared two or more injuries; four students did not report the number of injuries occurred. The first injury occurred, as an average, 17 days after the start of the internship (median 15 days) and, in 25% of the cases, during the first 9 days. The highest percentage of accidents occurred during the first internship (25.3% of the total) and decreased with the progress of the training path. The injuries occurred in 38% of cases during drug preparation, 24% when disposing of sharp devices, 15% while re-capping needles, 13% during blood sampling and 10% in other circumstances. In 51.2% of cases, the needle was not sterile. Among the nursing students who suffered a needle stick injury, 58.1% declared that they had performed the post-exposure prophylaxis. 96% of students stated to be vaccinated against Hepatitis B virus. Amongst the students who had their serological status checked (74%), 18% stated the antibody titre was not protective. 49.8% of students answered to have been trained in advance on the correct procedures to avoid needle stick and cutting edges injuries in each clinical ward attended, 41.2% referred that this occurred only in some wards and 10% in no ward at all. CONCLUSION: The results of this study show a high percentage of needle stick injuries in students of the Nursing Bachelor's Degree. Therefore, there is a need for careful reflection on the most effective methods of targeted training acquisition of knowledge, skills and behavioural models useful for the exercise of the profession.

Riflessioni della Water Decade 2005-215 e Giornate Mondiali dell'acqua

AIM: To evaluate the level of microbial contamination of air in operating theatres using active [i.e. surface air system (SAS)] and passive [i.e. index of microbial air contamination (IMA) and nitrocellulose membranes positioned near the wound] sampling systems. METHODS: Sampling was performed between January 2010 and January 2011 in the operating theatre of the orthopaedics department in a university hospital in Southern Italy. FINDINGS: During surgery, the mean bacterial loads recorded were 2232.9colony-forming units (cfu)/m(2)/h with the IMA method, 123.2cfu/m(3) with the SAS method and 2768.2cfu/m(2)/h with the nitrocellulose membranes. Correlation was found between the results of the three methods. Staphylococcus aureus was detected in 12 of 60 operations (20%) with the membranes, five (8.3%) operations with the SAS method, and three operations (5%) with the IMA method. CONCLUSION: Use of nitrocellulose membranes placed near a wound is a valid method for measuring the microbial contamination of air. This method was more sensitive than the IMA method and was not subject to any calibration bias, unlike active air monitoring systems.

This article described an outbreak of bacterial pneumonia/bronchopneumonia in a group of visitors to a mineral water spa contaminated with Pseudomonas aeruginosa in summer 2009. The epidemiologic investigation was activated after the hospitalization for pneumonia of three people who had undergone therapy at the spa. Discharge with a diagnosis of pneumonia/bronchopneumonia in weeks 27 to 39 of 2009 of four hospitals near the spa were data-linked with the list of spa’s visitors. Environmental samples of thermal water were performed and analysed. Investigations discovered a total of 39 cases of pneumonia among the spa visitors. Checks carried out in the hospitals near the spa revealed more hospitalizations for pneumonia in summer 2009 than the average for the previous five summers. There was a significant association between pneumonia and inhalation therapy (RR=7.33; p&lt;0.0001) and aerosol therapy (RR=8.25; p&lt;0.0001). P. aeruginosa was discovered in the water of the inhalation equipment. The spa micro-environment offers a risk of infection from pathogenic and/or opportunistic micro-organisms and therefore in-house testing, surveillance and prevention systems should be put in place for the well-being of clients.

The purpose of this study was to describe the susceptibility of environmental strains of Legionella spp. to 10 antimicrobials commonly used for legionellosis therapy. A study of environmental strains could be useful to timely predict the onset of antibiotic resistance in the environment before it is evidenced in clinical specimens.

Nosocomial yeast infections have increased significantly worldwide and especially in surgical and intensive care unit (ICU) patients. Although Candida species have various degrees of susceptibility to frequently used drugs, antifungal resistance is rare. A ten-year retrospective surveillance of candidemia was carried out in a University Hospital of Southern Italy. The aim of this study was the determination of Candida bloodstream infections (BSI) and central venous catheter (CVC)-related episodes, prevalence and in vitro susceptibility. 320 candidemia episodes were registered and 374 yeasts collected. Etest and Sensititre methods were used to test the isolates' susceptibility to amphotericin B, anidulafungin, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole. The results were compared with those of CLSI reference broth microdilution method. Most yeasts were susceptible to all antifungal drugs, with the exception of C. glabrata susceptibility to triazoles and C. tropicalis to fluconazole and voriconazole. As expectedC. parapsilosis isolates were generally associated with higher echinocandin minimum inhibitory concentrations (MICs) than the other Candida species. This study confirms the different antifungal susceptibility patterns among species, and underlines the need to perform antifungal susceptibility testing of clinically relevant yeasts.

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 72h, 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 48h and 72h 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>64mg/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 48h are proposed as optimal conditions for the evaluation of the in vitro antifungal susceptibility of M. pachydermatis using a modified CLSI BMD protocol.

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.

Despite being considered an emerging yeast related to immunocompromised individuals, severe infections by Malassezia furfur have not been evaluated. During a one-year survey on yeasts fungemia, 290 neonatal and 17 pediatric patients with intravascular catheters, lipid parenteral nutrition, prolonged ward stay, and surgery were enrolled. In addition, the origin of the infection was investigated by swabbing hand skin of patients, parents, and healthcare workers and medical devices. All biological specimens and swabs were cultured on Sabouraud dextrose agar and Dixon agar. The yeasts identification was based on morphological and biochemical features and by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and confirmed by sequencing the internal transcribed spacer of nuclear ribosomal DNA. A higher prevalence of M. furfur (2.1%) over Candida spp. (1.4%) caused bloodstream infections (BSIs). Twelve fungemia episodes were recorded: 2 by M. furfur in a pediatric ward and 10 in a neonatal intensive care unit (6 caused by M. furfur and 4 by Candida spp.). M. furfur was also isolated from the skin of all patients with BSIs, from the hand skin of a parent, and from an incubator surface and sheet. Patients with Candida spp. and M. furfur BSIs were successfully treated with intravenous liposomal Amphotericin B. These findings highlight the need for a more accurate etiological diagnosis in high-risk patients by adding lipid-supplemented culture media for Malassezia in the current mycological routine as the clinical features, patient management, and outcomes in both Candida and Malassezia fungemia do not differ.

Background: In the past several decades the number of individuals, particularly young adults, acquiring body art has increased very seriously in occidental society. In order to verify young adults’ practices and knowledge of the risks related to body art, an investigation was conducted among university freshmen in Bari (Apulia Region) and Naples (Campania Region), both in Southern Italy. Methods: The study was carried out during the period 2008-2010 through an anonymous questionnaire distributed to 3,368 people selected among 26 Degree Courses of the University of Bari and Naples Parthenope. The two regional samples were matched for age and sex. Results: On the whole, 84.4% declare to know the infectious risks associated with body art practices, but only 4.1% of them correctly identified the infectious diseases which can be transmitted through these procedures; while 59.2% of the sample declared that non-infectious diseases can occur after a tattoo or a piercing, but only 5.4% of them correctly identified allergies, cysts, bleeding and scars. The possible occurrence of infectious and non-infectious diseases has been acknowledged by most students in Apulia region (89.7 vs 79.1% and 64.8 vs 53.6% respectively). Of the 1,001 (32.4%) pierced and 597 (19.8%) tattooed youths 23.4% reported complications. Campania region youths reported a greater number of complications than Apulia did (30.3 vs 13.1%) and they also purchased more frequently their tattoos or piercings in unauthorized facilities (35.3% vs 15.9%). Conclusions: With regard to the knowledge of health risks associated to body art, it has been registered a higher awareness of infectious than non-infectious diseases. Information about freshmen's knowledge and practices could help in effective planning of health promotion strategies.

BACKGROUND: The practice of tattooing and piercing has expanded in western society. In order to verify young adults' knowledge of the risk and practices related to body art, an investigation was conducted among freshmen of the University of Bari in the region of Apulia, Italy. METHODS: The study was carried out in the Academic Year 2009-2010 through an anonymous self-administered written questionnaire distributed to 1.656 freshmen enrolled in 17 Degree Courses. RESULTS: Of the 1.598 students included in the analysis, 78.3% believe it is risky to undergo piercing/tattoo practices. AIDS was indicated as a possible infection by 60.3% of freshmen, hepatitis C by 38.2%, tetanus by 34.3% and hepatitis B by 33.7% of the sample. 28.1% of freshmen were not aware that there are also non-infectious complications. 29% of the sample had at least one piercing or tattoo (this percentage does not include earlobe piercing in women). Of those with body art, the decision to undergo body art was made autonomously in 57.9% of the participants. 56.3% of freshmen undergoing body art had taken less than a month to decide. With regard to the reasons that led the sample to undergo body art, 28.4% were unable to explain it, 23.8% answered to improve their aesthetic aspect, 18.4% to distinguish themselves from others, 12.3% for fashion; 17.1% for other reasons. 25.4% of the sample declared that they had a piercing (79.8% female vs 20.2% male; ratio M/F 1:4.0). The average age for a first piercing was 15.3 years (range 10-27; SD ± 2.9). 9.6% of the sample declared that they have a tattoo (69.9% female vs 30.1% male; ratio M/F 1:2.3). The average age for a first tattoo was 17.5 years (range 10-26, SD ± 2.4). CONCLUSIONS: Most of the freshmen knew about AIDS-related risks but not other potential risks. Body art is fairly common among young adults (especially women). The decision is often not shared with the family and is undertaken mostly without a specific reason or for the improvement of aesthetic aspect. Information about freshmen's knowledge, attitudes and practices could help in effective planning of health promotion strategies

Multiple-site colonization with Candida spp. is commonly recognized as a risk factor for invasive fungal infection in critically ill patients. We carried out a study to determine the relationship between Candida colonization and invasive infection in neurological patients admitted to an ICU. At admission (T0) and every three days for two weeks, different samples (pharynx swab, tracheal secretions, stomach contents, etc.) were collected for mycological surveillance. Candida mannan antigen and Candida anti-mannan antibodies were assayed. The Colonization Index (CI) and Corrected Colonization Index were calculated for each time point. Of all patients 70% was already colonized by Candida spp. at T0 and six of them had CI ≥ 0.5. Three patients developed candidemia; they had CI ≥ 0.5 before infection. Positive values of Candida mannan antigen and anti-mannan antibodies were found only in the patients with candidemia. The sensitivity and specificity of the Candida mannan test were 66.6% and 100%, respectively, while the sensitivity and specificity of the anti-mannan antibody test were 100%. In accordance with other authors, we find the surveillance cultures are useful to monitor the Candida colonization in ICU patients. In addition, the sequential observation of anti-mannan antibodies could contribute to early diagnosis of candidiasis more than Candida mannan antigen in immunocompetent patients.

BACKGROUND: Candida bloodstream infections (BSI) represent an important problem in Intensive Care Units (ICUs). The epidemiology of candidemia is changing with an increase in the proportion of Candida (C.) non-albicans. OBJECTIVES: An Italian 2-year observational survey on ICU was conducted to evaluate the species distribution and possible differences between BSI caused by C. albicans and C. non-albicans. For comparative purposes, we performed a European literature-based review to evaluate distribution and frequency of Candida spp. causing ICU candidemia, during the period 2000-2013. MATERIALS AND METHODS: This laboratorybased survey involved 15 microbiology centers (GISIA-3 study). All candidemia episodes in adult patients were considered. Data were prospectively collected from 2007 to 2008. PubMed was searched for peer-reviewed articles. RESULTS: In total, 462 candidemia episodes were collected. C. albicans accounted for 49.4% of the isolates, followed by C. parapsilosis (26.2%) and C. glabrata (10.4%). Mortality was higher in patients with C. non-albicans than C. albicans (47.3% vs. 32.4 %, p > 0.05). Among risk factors, parenteral nutrition was more common (p = 0.02) in non-albicans candidemia, while surgery was more frequent (p = 0.02) in C. albicans candidemia. Twenty-four relevant articles were identified. C. albicans was the predominant species in almost all studies (range 37.9% -76.3%). C. glabrata was commonly isolated in the German-speaking countries, France, UK and North Europe; C. parapsilosis in Turkey, Greece and Spain. CONCLUSIONS: Although C. non-albicans BSI is increasing, our study shows that C. albicans is still the predominant species in ICU candidemia. There are differences in the epidemiology of Candida BSI among European countries, with a prevalence of C. glabrata and C. parapsilosis in Northern and Southern countries, respectively.

Abstract Background/purpose: According 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 methods: The 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. Results: Caries prevalence was recorded in 38.3% of the sample. Estimated means and 95% confidence intervals of the DMFT index by gender were: 1.15 (0.91e1.39) for males, 1.26 (1.02e1.5) for females, and 1.21 (1.03e1.39) 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 (tZ3.125, PZ0.002), but significantly lower than the mean for south Italy (tZ2.125, PZ0.03). Results of the regression model showed that only the mother and father’s nationality and educational level contributed to the DMFT.

In the present work, we studied the distribution of Candida parapsilosis complex species and the antifungal susceptibility of clinical isolates collected during an Italian surveillance study of yeast invasive fungal infections (IFIs) in intensive care units (ICUs). Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution method. BanI digestion patterns of the secondary alcohol dehydrogenase polymerase chain reaction (PCR) products were used to identify C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis. A total of 138 C. parapsilosis isolates were stored (January 2007-December 2008). The overall frequency of C. parapsilosis complex in IFIs was 22%. Of the 138 tested isolates, 95% were C. parapsilosis sensu stricto, 3.6% were C. orthopsilosis, and 1.4% were C. metapsilosis. The MIC(50) values (expressed as μg/ml) for anidulafungin, caspofungin, and micafungin for C. parapsilosis complex were 2, 1, and 2, respectively, and the MIC(90) values were 4, 2, and 4, respectively. The MIC(50) and MIC(90) values for itraconazole and posaconazole were 0.12 and 0.25, respectively, and for fluconazole, they were 1 and 4, respectively. This study, the most comprehensive study conducted to date to evaluate the frequency and antifungal susceptibility profiles of C. parapsilosis complex isolates from critically ill patients in Italy, highlights the low prevalence of C. orthopsilosis and C. metapsilosis in IFIs.

Cryptococcus gattii has becoming more prevalent in temperate climate zones, during the past decades. We describe a C. gattii serotype B infection in an immunocompetent Italian patient with sclerosing cholangitis. The patient traveled once to Eastern Canada and otherwise no other countries than Italy were visited. Molecular analysis revealed that the C. gattii isolate belong to genotype AFLP4/VGI and has mating-type α which is the most common genotype in the Mediterranean environment. The C. gattii strain was found to be closely related, but not identical, to other C. gattii strains from the Mediterranean area

Healthcare-associated infections (HAIs) are an important issue in terms of quality of care. HAIs impact patient safety by contributing to higher rates of preventable mortality and prolonged hospitalizations. In Italy, analysis of the currently available accreditation systems shows a substantial heterogeneity of approaches for the prevention and surveillance of HAIs in hospitals. The aim of the present study is to develop and propose the use of a synthetic assessment tool that could be implemented homogenously throughout the nation.

BACKGROUND: Almonds are healthy nutraceuticals, which vary across different cultivars. We compared the composition, agreeability and gastrointestinal effects of two almond cultivars from different areas. METHODS: Californian Carmel (CAcv) and local Apulian Filippo Cea (FCcv) cultivars were compared for the chemical composition and sensory evaluation according to visual analogue and semiquantitative scales in 60 volunteers. Gallbladder/gastric motility (ultrasonography) and orocecal transit time (H2-breath test) were studied in another 24 subjects by comparing the effects of a standard liquid test meal with isovolumetric almond test meals (24 g of CAcv or FCcv almonds). RESULTS: Proteins prevailed in CAcv, while FCcv contained more lipids and 10-times more total phenol content than CAcv. For agreeability, CAcv scored higher than FCcv for smell, texture and appearance, although different perceptions existed in lean (scores for smell, taste, texture, appearance higher for CAcv than FCcv), obese (CAcv better than FCcv only for appearance) and elderly subjects (CAcv better than FCcv only for texture). Gallbladder emptying was stronger with FCcv than CAcv. Antral dilatation after ingestion of both cultivars was greater than the dilatation observed after the test meal. Gastric emptying, however, was similar after FCcv, CAcv and the test meal. The orocecal transit time in response to both cultivars was shorter than after the test meal. CONCLUSIONS: Differences in composition and effects of FCcv and CAcv cultivars support their potential use as valuable nutraceutical tools, to be confirmed in further clinical studies.

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, fungaemia and nosocomial outbreaks in neonates, children and adults and can be life-saving for those patients. Ma-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been reported to be 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. OBJECTIVES: To create an extensive database of main mass spectra (MSPs) that will allow quick identification of Malassezia species by MALDI-TOF MS. METHODS: 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. RESULTS: 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 of the ribosomal DNA. CONCLUSIONS: Implementation of the MALDI-TOF MS system as a routine identification tool will contribute to correct identification of Malassezia yeasts with minimal effort and in a short turnaround time, which is especially important for the rapid identification of Malassezia in skin diseases and nosocomial outbreaks.

According to Italian Ministerial Decree No. 185 of 12 June 2003, water is considered suitable for irrigation if levels of fecal bacteria (i.e., Escherichia coli and Salmonella) are within certain parameters. The detection of other microorganisms is not required. The aim of this study is to determine the bacteriological quality of groundwater used for irrigation and the occurrence of enteric viruses (Norovirus, Enterovirus, Rotavirus, Hepatovirus A), and to compare the presence of viruses with the fecal bacteria indicators. A total of 182 wells was analyzed. Widespread fecal contamination of Apulian aquifers was detected (141 wells; 77.5%) by the presence of fecal bacteria (i.e., E. coli, Salmonella, total coliforms, and enterococci). Considering bacteria included in Ministerial Decree No. 185, the water from 35 (19.2%) wells was unsuitable for irrigation purposes. Among 147 wells with water considered suitable, Norovirus, Rotavirus, and Enterovirus were detected in 23 (15.6%) wells. No Hepatovirus A was isolated. Consequently, 58 wells (31.9%) posed a potential infectious risk for irrigation use. This study revealed the inadequacy of fecal bacteria indicators to predict the occurrence of viruses in groundwater and it is the first in Italy to describe the presence of human rotaviruses in well water used for irrigation.

Data on the occurrence and epidemiology of Aspergillus spp. in laying hens farms are scant. With the aims of determining levels of airborne contamination in laying hen farms and evaluating the potential risk of infection for workers and animals, 57 air samples from 19 sheds (Group I), 69 from faeces (Group II), 19 from poultry feedstuffs (Group III) and 60 from three anatomical sites (i.e. nostrils, pharynx, ears) of 20 farm workers (Group IV) were cultured. The Aspergillus spp. prevalence in samples ranged from 31.6% (Group III) to 55.5% (Group IV), whereas the highest conidia concentration was retrieved in Group II (1.2 × 10(4) c.f.u. g(-1)) and in Group III (1.9 × 10(3) c.f.u. g(-1)). The mean concentration of airborne Aspergillus spp. conidia was 70 c.f.u. m(-3) with Aspergillus fumigatus (27.3%) being the most frequently detected species, followed by Aspergillus flavus (6.3%). These Aspergillus spp. were also isolated from human nostrils (40%) and ears (35%) (P<0.05) (Group IV). No clinical aspergillosis was diagnosed in hens. The results demonstrate a relationship between the environmental contamination in hen farms and presence of Aspergillus spp. on animals and humans. Even if the concentration of airborne Aspergillus spp. conidia (i.e. 70 c.f.u. m(-3)) herein detected does not trigger clinical disease in hens, it causes human colonization. Correct management of hen farms is necessary to control environmental contamination by Aspergillus spp., and could lead to a significant reduction of animal and human colonization.

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.

BACKGROUND: Candidemia has become an increasingly important problem in infants hospitalized in the Neonatal Intensive Care Units (NICUs). Candida species are the third most common agents of late-onset infections in critically ill neonates and they are associated with high morbidity and mortality rates. In this study we evaluated the epidemiology of Candida bloodstream infections in the NICU of an Italian university hospital during a 15-year period. Our specific aims were to analyze the change in species distribution and the vitro susceptibility of these yeasts to fluconazole (FCZ) and amphotericin B (AmB). METHODS: A retrospective study of candidemia in the NICU of a university hospital in southern Italy, covering the years 2000-2014 was carried out. The isolates were identified using the VITEK2 yeast identification system and antifungal susceptibility was determined using the E-test method. RESULTS: Among the 57 patients with confirmed candidemia, 60% were males (n = 34 cases) and 82% (n = 47) had a gestational age of 24-32 weeks. Twenty-seven neonates (47%) had a very low birth weight (<1500 g), 20 (35%) an extremely low birth weight (<1000 g), and 10 (18%) a low birth weight (<2500 g). The most important potential risk factors were the placement of a central venous catheter, total parenteral nutrition, and endotracheal intubation (100%, each). Candida albicans was the most frequent yeast (47%), followed by Candida parapsilosis (44%). The proportion of Candida non-albicans increased slightly, from 46% in 2000-2004 to 71% in 2010-2014 (χ2 test for trend, p = 0.030). All isolates were susceptible to FCZ and AmB. CONCLUSIONS: The detection in this epidemiologic study of an increase in Candida non-albicans highlights the importance of correct species-level identification in the rapid diagnosis for an efficient treatment of candidemia. Knowledge of the local epidemiological trends in Candida species isolated in blood cultures will facilitate therapeutic decision-making.

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.

Background. 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. Aim. To compare the ability of a dusting cloth pad (DC pad), with two other methods (Rodac contact plate and air sampling) to detect a contamination due to filamentous fungi (FF) in operating rooms (ORs). Methods. 110 sampling campaigns were carried out in ORs of hospitals of three Italian cities: Bari, Rome and Sassari. Surface samples were collected from the upper surface of scialytic lamps: half with two Rodac contact plates and other half was all rubbed with one DC pad and the dust captured was inoculated on a Petri plate. Next to the surgical table 540 liters of air were sampled, using SAS sampler. Air and Rodac plates sampling follow standard methods for detecting fungi. Finding. Overall, 96% (90/94) of the DC pad samples were positive compared with 51% (48/94) of Rodac plates (p<0.0001) and 35% (33/94) of air samples (p<0.0001). The difference persists stratifying by venue, with the exception of Sassari, due to a high percentage of positive samples with all methods. Conclusion. The DC pad improves the ability to detect an environmental contamination of FF.

Abstract: Healthcare facilities (HF) represent an at-risk environment for legionellosis transmission occurring after inhalation of contaminated aerosols. In general, the control of water is preferred to that of air because, to date, there are no standardized sampling protocols. Legionella air contamination was investigated in the bathrooms of 11 HF by active sampling (Surface Air System and Coriolis®) and passive sampling using settling plates. During the 8-hour sampling, hot tap water was sampled three times. All air samples were evaluated using culture-based methods, whereas liquid samples collected using the Coriolis® were also analyzed by real-time PCR. Legionella presence in the air and water was then compared by sequence-based typing (SBT) methods. Air contamination was found in four HF (36.4%) by at least one of the culturable methods. The culturable investigation by Coriolis® did not yield Legionella in any enrolled HF. However, molecular investigation using Coriolis® resulted in eight HF testing positive for Legionella in the air. Comparison of Legionella air and water contamination indicated that Legionella water concentration could be predictive of its presence in the air. Furthermore, a molecular study of 12 L. pneumophila strains confirmed a match between the Legionella strains from air and water samples by SBT for three out of four HF that tested positive for Legionella by at least one of the culturable methods. Overall, our study shows that Legionella air detection cannot replace water sampling because the absence of microorganisms from the air does not necessarily represent their absence from water; nevertheless, air sampling may provide useful information for risk assessment. The liquid impingement technique appears to have the greatest capacity for collecting airborne Legionella if combined with molecular investigations

Legislators have implemented policies to improve food labelling to protect consumers and to make the presentation of ingredients and nutritional information more transparent. Proper food labelling allows consumers who may suffer from food allergies or intolerances to know exactly what ingredients a product contains, and it also helps them make more informed health and nutrition choices. This paper deals with the most current European and Italian legislation on food labelling, actions taken in non-EU countries to increase health choices, and the expected impact on Public Health.

We determined the in vitro antifungal activity of liposomal amphotericin B (L-AmB) against 604 clinical yeast isolates. Amphotericin B deoxycholate (D-AmB) was tested in parallel against all the isolates. Susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) M27-A3 method. Overall, L-AmB was highly active against the isolates (mean MIC = 0.42 μg/ml; MIC90= 1μg/ml; 97.2% of MICs were ≤ 1μg/ml) and comparable to D-AmB (mean MIC = 0.48μg/ml; MIC90= 1μg/ml; 97.3% of MICs were ≤ 1μg/ml). The in vitro activity of D-AmB and L-AmB was correlated (R2 = 0.61; exp (Coef.) = 2.3; 95% IC= 2.19-2.44, p&lt;0.001). Candida albicans (mean MICs of D-AmB and L-AmB, 0.39 μg/ml and 0.31 μg/ml, respectively) and Candida parapsilosis (mean MICs of D-AmB and L-AmB, 0.38 μg/ml and 0.35 μg/ml, respectively) were the species most susceptible to the agents tested, while Candida krusei (currently named Issatchenkia orientalis) (mean MICs of D-AmB and L-AmB, 1.27 μg/ml and 1.13 μg/ml, respectively) was the least susceptible. The excellent in vitro activity of L-AmB may have important implications for empirical treatment approaches and support its role in treatment of a wide range of invasive infections due to yeasts

Although guidelines for the treatment of Malassezia furfur fungemia are not yet defined, clinical data suggest that amphotericin B (AmB) is effective for treating systemic infections. In the absence of clinical breakpoints for Malassezia yeasts, epidemiological cut-off values (ECVs) are useful to discriminate between isolates with and without drug resistance. This study aimed to compare the distribution of minimal inhibitory concentration (MIC) and the ECVs for AmB of both deoxycholate (d-AmB) and liposomal (l-AmB) formulations of M. furfur isolates. The 84 M. furfur strains analyzed, which included 56 from blood, sterile sites and catheters, and 28 from skin, were isolated from patients with bloodstream infections. MICs were determined by the modified broth microdilution method of the Clinical and Laboratory Standards Institute (CLSI). The l-AmB MIC and the ECVs were two-fold lower than those of d-AmB and a lower l-AmB mean MIC value was found for blood isolates than from skin. The ECVs for l-AmB and d-AmB were 8 mg/l and 32 mg/l, respectively. Three strains (3.6%) showed l-AmB MIC higher than ECV (MIC > 8mg/l) of which two were isolated from the catheter tip of patients treated with micafugin, l-Amb and fluconazole, and one from skin. The results showed that the l-AmB might be employed for assessing the in vitro antifungal susceptibility of M. furfur by a modified CLSI protocol and that ECVs might be useful for detecting the emergence of resistance.

The antifungal susceptibilities of 598 isolates of Candida spp. (bloodstream, other sterile sites) to liposomal amphotericin B (L-AmB) versus amphotericin B (AmB) were determined. Minimal inhibitory concentrations (MICs) were calculated using the Clinical and Laboratory Standards Institute broth microdilution (BMD; M27-A3) method for L-AmB and the E-test method for AmB. The MIC50/MIC90 (µg ml-1) values for L-AmB BMD and AmB E-test were 0.25/1 and 0.19/0.5, respectively. The overall essential agreement (± 2 dilutions) was 91.5%, ranging from 37.5% (Candida lusitaniae) to 100% (Candida glabrata and Candida krusei). Categorical agreement (CA) between the two methods was categorized on the basis of previously published breakpoint (susceptible/resistant MIC cut-off of 1 µg ml-1). The overall CA at the 48-h reading was 97.3%, ranging from 72.7% (C. krusei) to 100% (Candida albicans). Major and very major discrepancies occurred in 2.3% and 0.3%, respectively. Spearman's rho was 0.48 (p < 0.0001). These results demonstrate the utility of the AmB E-test as a surrogate marker to predict the sensibility and resistance of Candida species to L-AmB, and thus to support its use in antifungal treatment.

Fungaemia caused by Malassezia spp. in hospitalized patients requires prompt and appropriate therapy, but standard methods for the definition of the in vitro antifungal susceptibility have not been established yet. In this study, the in vitro susceptibility of Malassezia furfur from bloodstream infections (BSIs) to amphotericin B (AMB), fluconazole (FLC), itraconazole (ITC), posaconazole (POS) and voriconazole (VRC) was assessed using the broth microdilution (BMD) method of the Clinical and Laboratory Standards Institute (CLSI) with different media such as modified Sabouraud dextrose broth (SDB), RPMI and Christensen's urea broth (CUB). Optimal broth media that allow sufficient growth of M. furfur, and produce reliable and reproducible MICs using the CLSI BMD protocol were assessed. Thirty-six M. furfur isolates collected from BSIs of patients before and during AMB therapy, and receiving FLC prophylaxis, were tested. A good growth of M. furfur was observed in RPMI, CUB and SDB at 32 °C for 48 and 72 h. No statistically significant differences were detected between the MIC values registered after 48 and 72 h incubation. ITC, POS and VRC displayed lower MICs than FLC and AMB. These last two antifungal drugs showed higher and lower MICs, respectively, when the isolates were tested in SDB. SDB is the only medium in which it is possible to detect isolates with high FLC MICs in patients receiving FLC prophylaxis. A large number of isolates showed high AMB MIC values regardless of the media used. In conclusion, SDB might be suitable to determine triazole susceptibility. However, the media, the drug formulation or the breakpoints herein applied might not be useful for assessing the AMB susceptibility of M. furfur from BSIs.

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.

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 1C 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.

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.

In this paper the authors investigated a synergistic antimycotic effect between four antifungal drugs Ampho- tericin 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 Anidula- fungin 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 an- timycotic efficacy was obtained from the association between Anidulafungin and Amphotericin B or Fluconazole (Mix- ture 4:6). In contrast the association Tioconazole/Anidulafungin was less effective on fungal species growth. The antimy- cotics MIC reduction values were more evident against some strains as C. glabrata, C. krusei, C. tropicalis and C. parap- silosis.

The aim of this multicenter prospective study was to evaluate the incidence of invasive fungal infections (IFIs) in adult and pediatric patients with hematologic malignancies, involving nine nosocomial facilities in Southern Italy over a period of 18 months. Furthermore, results of an environmental microbial surveillance routinely carried out in some of the enrolled hospitals are reported. A total of 589 oncohematological patients were enrolled and 27 IFIs were documented. The main infections were caused by yeasts, more than filamentous fungi (overall incidence of 2.7% and 1.9%, respectively). The yeasts were mainly represented by Candida spp. (87.5%), all isolated by blood cultures; C. parapsilosis was the most common species. Among mould infections, the most frequent site was the lung, with regard to aspergillosis (81.8%). In six of the 10 patients with suspected aspergillosis, the diagnosis was made by the detection of galactomannan and (1,3)-β-D-glucan antigens. The microbiological surveillance carried out on 156 air, 312 water and 312 surface samples revealed low environmental contamination: Alternaria alternata was the only fungus isolated from two surface samples. Our data, especially the low occurrence of filamentous fungi, suggest a particular local epidemiology. Further studies are needed to confirm this microbiological trend in onco-hematological patients in Southern Italy, the results of which might be helpful to improve the management of these patients.

Introduction. During the past years invasive fungal infections (IFIs) have become an increasingly important problem in infants hospitalized in the Neonatal Intensive Care Unit (NICU). Candida species is the third most-common agent of late-onset infections in critically ill neonates, with an estimated incidence of 2.6-10% in very low birth weight and 5.5-20% in extremely low birth weight infants. The aim of this observational study is to evaluate the epidemiology of IFIs among infants admitted to NICUs of one Italian region by a multicenter surveillance (Aurora Project). Methods. The IFIs surveillance was carried out prospectively in Apulia (Southern Italy) between February 2007 and August 2008. This report focuses on the results from 6 enrolled NICUs. Results. Twenty-one neonates developed IFIs: the overall incidence was 1.3% and crude mortality was 23.8%. Infants weighing ≤ 1500 g (4.3%) showed a significantly higher incidence than those ≥ 2500 g (0.2%). C.parapsilosis (61.9%) was the most frequent isolated species. The main potential risk factors were having a central venous catheter placed, length of stay in NICU > 7 days and total parenteral nutrition for > 5 days. The (1,3)-β-D glucan (BDG), mannan antigens and anti-Candida antibodies' evaluation was performed in 7 neonates. All neonates were positive to the BDG; the mannan antigen result was positive in 5 newborns, the anti-mannan antibodies were always negative. All isolates were amphotericin B and fluconazole-susceptible. Discussion. This first prospective study on neonatal fungal infection in one Italian region gives evidence of a preponderance of non-albicans Candida spp and indicates potential utility of BDG as an adjunct diagnostic test.

The dental practice is associatedwith a high risk of infections, both for patients and healthcare operators, and the environment may play an important role in the transmission of infectious diseases. A microbiological environmental investigation was carried out in six dental clinics as a pilot study for a larger multicentre study that will be performed by the Italian SItI (Society of Hygiene, Preventive Medicine and Public Health) working group &quot;Hygiene in Dentistry&quot;. Microbial contamination of water, air and surfaces was assessed in each clinic during the five working days of the week, before and during treatments. Air and surfaceswere also examined at the end of the daily activity. A wide variation was found inmicrobial environmental contamination, both within the participating clinics and relative to the different sampling times. Microbial water contamination in Dental Unit Water Systems (DUWS) reached values of up to 26×104 cfu/mL (colony forming units per millilitre). P. aeruginosa was found in 33% of the sampled DUWS and Legionella spp. in 50%. A significant decrease in the Total Viable Count (TVC) was recorded during the activity. Microbial air contamination showed the highest levels during dental treatments and tended to decrease at the end of the working activity (pb0.05). Microbial buildup on surfaces increased significantly during the working hours. As these findings point out, research on microbial environmental contamination and the related risk factors in dental clinics should be expanded and should also be based on larger collections of data, in order to provide the essential knowledge aimed at targeted preventive interventions.

BACKGROUND: Traumatic dental injuries occur frequently in children and adolescents. The purpose of the present study is to examine the levels of knowledge and behaviors regarding dental trauma among parents of children attending primary schools in the Apulia region of Italy. METHODS: The study was carried out using an anonymous questionnaire with closed answers distributed to 2,775 parents who were enrolled based on the entire regional school population. Analyses were conducted using the PROC CORRESP (procedure to perform multiple correspondence analysis) and PROC FASTCLUS (procedure to perform cluster analysis). Statistical significance was set at p-value <0.05. RESULTS: A total 15.5% of the sample reported that their children had experienced dental trauma. Overall, 53.8% of respondents stated that they knew what to do in cases of dental injury. Regarding the time limit within which it is possible to usefully intervene for dental trauma, 56.8% of respondents indicated "within 30 minutes". Of the total sample, 56.5% knew how to preserve a displaced tooth. A total 62.9% of parents felt it was appropriate for their children to use dental guards during sports activities. The multivariate analysis showed that wrong knowledge are distributed among all kinds of subject. Parents with previous experience of dental trauma referred right behaviours, instead weak knowledge and wrong behaviours are associated with parents that easily worried for dental events. CONCLUSIONS: This study showed that most parents reported no experience of dental trauma in their children, and half of them did not know what to do in case of traumatic dental injury and they would intervene within 30 minutes, suggesting that dental trauma may trigger panic. However, they did not have the information needed to best assist the affected child. Motivating parents to assume a preventive approach towards dental trauma may produce positive changes that would result an increase of long-term health benefits among both parents and children.

Background: Following the publication of the Italian Guidelines for the control and prevention of legionellosis an environmental and clinical surveillance has been carried out in Southeastern Italy. The aim of the study is to identify the risk factors for the disease, so allowing better programming of the necessary prevention measures. Methods: During the period January 2000 - December 2009 the environmental surveillance was carried out by water sampling of 129 health care facilities (73 public and 56 private hospitals) and 533 buildings within the community (63 private apartments, 305 hotels, 19 offices, 4 churches, 116 gyms, 3 swimming pools and 23 schools). Water sampling and microbiological analysis were carried out following the Italian Guidelines. From January 2005, all facilities were subject to risk analysis through the use of a standardized report; the results were classified as good (G), medium (M) and bad (B). As well, all the clinical surveillance forms for legionellosis, which must be compiled by physicians and sent to the Regional Centre for Epidemiology (OER), were analyzed. Results: Legionella spp. was found in 102 (79.1%) health care facilities and in 238 (44.7%) community buildings. The percentages for the contamination levels &lt; 1,000, 1,000-10,000, &gt; 10,000 cfu/L were respectively 33.1%, 53.4% and 13.5% for samples from health care facilities and 33.5%, 43.3% and 23.2% for samples from the community. Both in hospital and community environments, Legionella pneumophila serogroup (L. pn sg) 2-14 was the most frequently isolate (respectively 54.8% and 40.8% of positive samples), followed by L. pn sg 1 (respectively 31.3% and 33%). The study showed a significant association between M or B score at the risk analysis and Legionella spp. positive microbiological test results (p &lt; 0.001). From clinical surveillance, during the period January 2001 - August 2009, 97 cases of legionellosis were reported to the OER: 88 of community origin and 9 nosocomial. The most frequent symptoms were: fever (93.8%), cough (70.1%), dyspnea (58.8%), shivering (56.7%). Radiological evidence of pneumonia was reported in 68%. The laboratory diagnostic methods used were: urinary antigen (54.3%), single antibody titer (19.8%), only seroconversion (11.1%), other diagnostic methods (14.8%). Conclusions: Our experience suggests that risk analysis and environmental microbiological surveillance should be carried out more frequently to control the environmental spread of Legionella spp. Furthermore, the laboratory diagnosis of legionellosis cannot be excluded only on the basis of a single negative test: some patients were positive to only one of the diagnostic tests.

This study represents the first investigation of microbiological groundwater pollution as a function of aquifer type and season for the Apulia region of southern Italy. Two hundred and seven wells were randomly selected from those monitored by the Regional Agency for Environmental Protection for emergency use. Both compulsory (Escherichia coli, Total Coliform, and Enterococci) and optional (Pseudomonas aeruginosa, Salmonella spp., Heterotrophic Plate Count at 37 and 22 °C) microbiological parameters were assessed regularly at these wells. Groundwater from only 18 of the 207 (8.7 %) wells was potable; these all draw from karst-fissured aquifers. The remaining 189 wells draw from karst-fissured (66.1 %) or porous (33.9 %) aquifers. Of these, 82 (43.4 %) tested negative for Salmonella spp. and P. aeruginosa, while 107 (56.6 %) tested positive for P. aeruginosa (75.7 %), Salmonella spp. (10.3 %), or for both Salmonella spp. and P. aeruginosa (14 %). A logistic regression model shows that the probability of potable groundwater depends on both season and aquifer type. Typically, water samples were more likely to be potable in autumn-winter than in spring-summer periods (odds ratio, OR = 2.1; 95 % confidence interval, 95 % CI = 1.6-2.7) and from karst-fissured rather than porous aquifers (OR = 5.8; 95 % CI = 4.4-7.8). Optional parameters only showed a seasonal pattern (OR = 2.6; 95 % CI = 1.7-3.9). Clearly, further investigation of groundwater microbiological aspects should be carried out to identify the risks of fecal contamination and to establish appropriate protection methods, which take into account the hydrogeological and climatic characteristics of this region.

A microbiological environmental investigation was carried out in ten dental clinics in Italy. Microbial contamination of water, air and surfaces was assessed in each clinic during the five working days, for one week per month, for a three-month period. Water and surfaces were sampled before and after clinical activity; air was sampled before, after, and during clinical activity. A wide variation was found in microbial environmental contamination, both within the participating clinics and for the different sampling times. Before clinical activity, microbial water contamination in tap water reached 51,200cfu/mL (colony forming units per milliliter), and that in Dental Unit Water Systems (DUWSs) reached 872,000cfu/mL. After clinical activity, there was a significant decrease in the Total Viable Count (TVC) in tap water and in DUWSs. Pseudomonas aeruginosa was found in 2.38% (7/294) of tap water samples and in 20.06% (59/294) of DUWS samples; Legionella spp. was found in 29.96% (89/297) of tap water samples and 15.82% (47/297) of DUWS samples, with no significant difference between pre- and post-clinical activity. Microbial air contamination was highest during dental treatments, and decreased significantly at the end of the working activity (p<0.05). The microbial buildup on surfaces increased significantly during the working hours. This study provides data for the establishment of standardized sampling methods, and threshold values for contamination monitoring in dentistry. Some very critical situations have been observed which require urgent intervention. Furthermore, the study emphasizes the need for research aimed at defining effective managing strategies for dental clinics.

The Candida parapsilosis group encompasses three species: C. parapsilosis, C. orthopsilosis, and C. metapsilosis. Here, we describe the incidence and echinocandin susceptibility profile of bloodstream isolates of these three species collected from patients admitted to an Italian university hospital from 2007 to 2014. Molecular identification of cryptic species of the C. parapsilosis complex was performed using polymerase chain reaction amplification of the gene encoding secondary alcohol dehydrogenase, followed by digestion with the restriction enzyme BanI. Minimum inhibitory concentrations were determined using the broth microdilution method according to European Committee for Antimicrobial Susceptibility Testing (EUCAST EDef 7.2) and Clinical Laboratory Standards Institute (CLSI M27-A3) guidelines, and the results were compared with those obtained using the E-test and Sensititre methods. Of the 163 C. parapsilosis complex isolates, 136 (83.4%) were identified as C. parapsilosis, and 27 (16.6%) as C. orthopsilosis. The species-specific incidences were 2.9/10,000 admissions for C. parapsilosis and 0.6/10,000 admissions for C. orthopsilosis. No resistance to echinocandins was detected with any of the methods. The percent essential agreement (EA) between the EUCAST and E-test/Sensititre methods for anidulafungin, caspofungin, and micafungin susceptibility was, respectively, as follows: C. parapsilosis, 95.6/97.8, 98.5/88.2, and 93.4/96.3; C. orthopsilosis, 92.6/92.6, 96.3/77.8, and 63.0/66.7. The EA between the CLSI and E-test/Sensititre methods was, respectively, as follows: C. parapsilosis, 99.3/100, 98.5/89.0, and 96.3/98.5; C. orthopsilosis, 96.3/92.6, 100/81.5, and 92.6/88.9. Only minor discrepancies, ranging from 16.9% (C. parapsilosis) to 11.1% (C. orthopsilosis), were observed between the CLSI and E-test/Sensititre methods. In conclusion, this epidemiologic study shows a typical C. parapsilosis complex species distribution, no echinocandin resistance, and it reinforces the relevance of using commercially available microbiological methods to assess antifungal susceptibility. These data improve our knowledge of the national distribution of species of the psilosis group, as there are very few studies of these species in Italy

Mycotoxins are secondary metabolites produced by several species of fungi and characterized by a range of dangerous health effects for humans and animals. The reference regulations in this field are continuously improved and updated worldwide. This paper reports the state of the art of Italian legislation, in the European context, concerning the limits of some mycotoxins in foodstuffs and the procedures for performing official controls.

Legionellae are opportunistic bacteria that cause various conditions after exposure to contaminated aerosols, ranging from a serious type of pneumonia to a mild case of an influenza-like illness. Despite the risks of exposure, little is known about the occurrence of Legionella in natural environments and, even though studies have shown that there is a potential risk of transmission via inhalation, it does not have to be detected in groundwater that is used for irrigation. The culture methods traditionally used to detect Legionella have several limits that can be partly solved by applying molecular techniques. Samples from 177 wells in Apulia, Southern Italy, were collected twice, in winter and in summer, and analyzed. When compared with the guidelines, 145 (81.9%) of the sampled wells were suitable for irrigation use. The culture-based method highlighted the presence of different species and serogroups of Legionella in 31 (21.2%) of the 145 wells that were shown to be suitable for irrigation use. A greater number of wells returned positive results for Legionella in summer than in winter (p = 0.023), and the median concentrations were mostly higher in summer (500 CFU/L) than in winter (300 CFU/L). The median temperature in the Legionella positive well waters was significantly higher than that in the negative ones, both in winter and in summer (p < 0.001). Using molecular techniques, Legionella non-pneumophila was found in 37 of the 114 wells earlier detected as suitable for irrigation use but negative for Legionella by the culture-based methods. The distribution of Legionella differ significantly in porous aquifers compared to the karst-fissured ones both with quantitative polymerase chain reaction (qPCR) (p = 0.0004) and viable cells by propidium monoazide (PMA-qPCR) (p = 0.0000). Legionella concentrations were weakly correlated with temperature of water both with qPCR (ρ = 0.47, p = 0.0033) and PMA-qPCR (ρ = 0.41, p = 0.0126). Our data suggest that water that aerosolizes when sprinkled on plants represents a potential source of Legionellosis, with a higher risk from exposure in summer. On a practical level, this finding is important for workers (farmers and gardeners) who are in contact with waters used for irrigation.

Methicillin-resistant S. aureus (MRSA) is a pathogen spread not only in the hospital environment but also in the community and amongst livestock (LA-MRSA). LA-MRSA can be transmitted to humans that live in close contact with MRSA-colonized animals, and human colonization and/or infection has been reported worldwide, particularly among those involved with livestock farming. In this study the authors evaluated the prevalence of S. aureus and MRSA among healthy carriers who worked in the food industry in Apulia, Southern Italy.

We would like to thank Collins andWalker for their comments and for acknowledging that this is an area requiring more research to improve our basic understanding of Legionella [1]. [...].

OBJECTIVE: This study aim was to determine the prevalence of microorganisms in the respiratory tract of patients with cystic fibrosis (CF) admitted to the CF Reference Centre in Southern Italy between 2002-2010. METHODS: Microbiology assessment of samples (sputum and tracheal aspirates) collected from patients with pulmonary exacerbation admitted to hospital was carried out. All patients were registered in a database and clinical and microbiological data were retrospectively analysed. RESULTS: Overall, 188 patients were included and a total of 1217 samples were analysed. The most common microorganisms were Staphylococcus aureus (78.7% of the patients) and Pseudomonas aeruginosa (58%), followed by Candida albicans (19.1%), Haemophilus influenzae (13.3%) and Aspergillus fumigatus (9.6%). CONCLUSION: Compared to similar studies performed in other European countries, our microbiological data, especially the low occurrence of filamentous fungi, suggest a specific local epidemiology, probably related to some uncommon CFTR mutations, which are specific to Southern Italy.

Objective. The present report aims to molecularly characterize seven clinical L. pneumophila (L. pn.) sg 1 isolated from nosocomial acquired infections in Apulia region, using the European Working Group on Legionella Infections (EWGLI), sequence-based typing (SBT) and amplified fragment length polymorphism (AFLP) protocols and to compare the identified sequence types (STs) with those available in the EWGLI database. Methods. In the period, January 2000-December 2012, 151 cases (136 of community and 15 of nosocomial origin) of Legionnaires' disease were notified to the Regional Center for Epidemiology. With regard to nosocomial cases, 8 were confirmed by the isolation of Legionella spp. from respiratory secretions. These clinical isolates were characterized by amplified fragment length polymorphism (AFLP) and sequence-based typing (SBT), using the EWGLI standardized protocol. Results. The clinical isolates belong to ST42, ST23 and ST1. The AFLP confirms the SBT results. Comparing the STs herein detected with those already in the EWGLI SBT database, the 3 STs are frequent in other European countries. Conclusions. The molecular analysis demonstrates that the 3 STs are the most frequent in Italy and in Europe, supporting the hypothesis that some specific L. pn. sg 1 clones have gained widespread dissemination probably due to a common ecological niche. Further researches are required to investigate the potential changing incidence of STs and the fitness of emerging strains or clonal groups in environmental strains

Background: Legionella is anintracellularmicroorganismlivinginnaturalandartificial aquaticen- vironments. Althoughitstransmissiontohumansislinkedtotheinhalationofcontaminatedaerosols, there isnovalidatedairsamplingmethodforthecontrolandpreventionofthedisease.Theaimofthe present studywastoprovidemoreinformationonthedistributionof Legionella spp. inindooren- vironments andtodeterminewhetherthesame Legionella strains areisolatedfromairandwater samples. Methods: TenhealthcarefacilitieslocatedinsevenregionsofItalywereenrolled.Theserologicaltyping of Legionella spp. fromwatersamplesandthesurroundingairbyactiveandpassivesamplingwasas- sessed usingpolyvalentandmonovalentantisera.Subsequently,thestrainsidentified as Legionella pneumophila (Lpn) underwentmoleculartypingbysequence-basedtyping(SBT)usingsevengenes(flaA, pilE, asd, mip, mompS, proA, and neuA). Theallelicprofile numberwasassignedusingtheEuropean WorkingGroupfor Legionella Infections–SBT database. Results: Lpn serogroup6wasthemostprevalentserogroup;itwasfoundsimultaneouslyintheairand watersamplesofthreedifferenthealthcarefacilities.Intheremainingsevenhospitals, Lpn serogroups1, 6, 7,9,and12wereisolatedexclusivelyfromwatersamples.Themolecularinvestigationshowedthat Lpn strains inthewaterandairsamplesofeachpositivehealthcarefacilityhadthesameallelicprofile. Strains,identified assequencetypes(STs)728andST1638þST 1324,wereisolatedintworespective healthcare facilities,andanewstrain,identified asST1989,wasobtainedinonehealthcarefacility

PURPOSE: We compared the risk factors, the diagnostic tools and the outcome of filamentous fungal infections (FFIs) in hematological patients (HAEs) and non-hematological patients (non-HAEs). METHODS: Prospective surveillance (2009-2011) of proven and probable FFIs was implemented in 23 Italian hospitals. RESULTS: Out of 232 FFIs, 113 occurred in HAEs and 119 in non-HAEs. The most frequent infection was invasive aspergillosis (76.1 % for HAEs, 56.3 % for non-HAEs), and the localization was principally pulmonary (83.2 % for HAEs, 74.8 % for non-HAEs). Neutropenia was a risk factor for 89.4 % HAEs; the main underlying condition was corticosteroid treatment (52.9 %) for non-HAEs. The distribution of proven and probable FFIs was different in the two groups: proven FFIs occurred more frequently in non-HAEs, whereas probable FFIs were correlated with the HAEs. The sensitivity of the galactomannan assay was higher for HAEs than for non-HAEs (95.3 vs. 48.1 %). The overall mortality rate was 44.2 % among the HAEs and 35.3 % among the non-HAEs. The etiology influenced the patient outcomes: mucormycosis was associated with a high mortality rate (57.1 % for HAEs, 77.8 % for non-HAEs). CONCLUSIONS: The epidemiological and clinical data for FFIs were not identical in the HAEs and non-HAEs. The differences should be considered to improve the management of FFIs according to the patients' setting.

Nelle strutture termali diversi fattori, ad es. la temperatura delle acque impiegate, possono favorire la moltiplicazione di microrganismi come Legionella spp. La tipologia di utenza che frequenta queste strutture è spesso rappresentata anche da soggetti a rischio infettivo che si sottopongono a terapia inalatoria e a trattamenti in vasche idromassaggio. Di conseguenza, in mancanza di un'attenta gestione degli impianti, questi ambienti possono rappresentare una possibile sorgente di infezione da Legionella spp. Scopo del presente studio è verificare lo stato dell'arte della normativa italiana e pugliese che regola il controllo della contaminazione da Legionella spp. nelle strutture termali.

In most regions of the world, safeguarding groundwater resources is a serious issue, particularly in coastal areas where groundwater is the main water source for drinking, irrigation and industry. Water availability depends on climate, topography and geology. The aim of this paper is to evaluate aquifer recharge as a possible strategy to relieve water resource scarcity. Natural aquifer recharge is defined as the downward flow of water reaching the water table, increasing the groundwater reservoir. Hydro-meteorological factors (rainfall, evapotranspiration and runoff) may alter natural recharge processes. Artificial aquifer recharge is a process by which surface water is introduced with artificial systems underground to fill an aquifer. As a consequence of global warming that has increased the frequency and severity of natural disasters like the drought, the impacts of climate change and seasonality, the artificial recharge has been considered as a viable option. Different direct and indirect techniques can be used, and the choice depends on the hydrologic characteristics of a specific area. In Italy, Legislative Decree no. 152/06 plans artificial aquifer recharge as an additional measure in water management, and Decree no. 100/2016 establishes quantitative and qualitative conditions for recharge. Many projects examine aquifer recharge, such us WADIS-MAR in the southern Mediterranean region, WARBO in Italy and municipal wastewater treatment project in Apulia, a southern Italian region. However, aside from groundwater recharge, the community must foster a spirit of cooperation to manage groundwater as a sustainable resource.

Studies of meningococcal carriage are essential in improving knowledge of the epidemiology of meningococcal disease. The aim of this study is to ascertain the carrier rate and the serogroups of Neisseria Meningitidis circulating in a sample of students from the University of Bari. The population consisted of university students from the University of Bari - School of Medicine, who were invited to take a nasopharyngeal swab. The swabs were plated on selective plate medium; cultural and MLST tests were performed. Of 583 university students 12 carriers were identified (2%). 9 isolates proved auto-agglutinable. The other strains belonged to serogroups B, W135 and Y. Auto-agglutinable strains belonged to different clonal complexes, of which ST-53 was the most common. Only one strain, that belonged to ST-23/cluster A3 clonal complex, could cause meningococcal disease. No type C serogroup strain was detected and this could be directly related to immunization policies that provided meningococcal serogroup C conjugate vaccines for newborns and adolescents. The changing pattern of circulating serogroups of Neisseria meningitidis in healthy carriers could support a new immunization strategy which could provide quadrivalent meningococcal conjugate vaccines to pre-adolescents and adults.