assessing natural mineral water microbiology quality in the absence of cultivable pathogen bacteria

Abstract Italian Directives recommend the good quality of natural mineral waters but literature data assert a potential risk from microorganisms colonizing wellsprings and mineral water bottling plants. We evaluated the presence of microorganisms in spring waters (SW) and bottled mineral waters (BMW) samples. Routine microbiological indicators, additional microorganisms like Legionella spp., Nontuberculous mycobacteria (NTM) and amoebae (FLA) were assessed in 24 SW and 10 BMW samples performing cultural and molecular methods. In 33 out of 34 samples, no cultivable bacteria ≥10 CFU/L was found. Cultivable FLA were detected in 50% of water samples. qPCR showed the presence of Legionella qPCR units in 24% of samples (from 1.1 × 102 to 5.8 × 102 qPCR units/L) and NTM qPCR units in 18% of samples (from 1 × 102 to 1 × 105 qPCR units/L). Vermamoeba vermiformis and Acanthamoeba polyphaga were recovered respectively in 70% of BMW samples (counts from 1.3 × 103 to 1.2 × 105 qPCR units/L) and 42% of SW samples (from 1.1 × 103 to 1.3 × 104 qPCR units/L). Vahlkampfia spp. was detected in 42% of SW and 70% of BMW samples (from 1.2 × 103 to 1.2 × 105 qPCR units/L). Considering the presence of FLA, we underline the importance of a wider microbiological risk assessment in natural mineral waters despite the absence of cultivable bacteria

Colonization by Legionella spp. of water networks in residential buildings of the Province of Pisa, Italy

Despite the increase of community acquired cases of legionellosis in Italy over the last years, the Italian guidelines do not give indications for prevention and control of Legionella in the hot water networks (or centralized conditioning systems) of residential buildings. We performed a survey on eight medium sized apartment buildings in the Pisa district to assess the prevalence of Legionella spp. in the water network and the respondance to drinking water requisites at the point of use, according to the Italian norms

Presence of Legionella spp. in Hot Water Networks of Different Italian Residential Buildings: A Three-Year Survey

Although the European reports highlight an increase in community-acquired Legionnaires' disease cases, the risk of Legionella spp. in private houses is underestimated. In Pisa (Italy) we performed a three-year survey on Legionella presence in 121 buildings with an independent hot water production (IB); 64 buildings with a central hot water production (CB); and 35 buildings with a solar thermal system for hot water production (TB). From all the 220 buildings Legionella spp. was researched in two hot water samples collected either at the recirculation point or on the first floor and on the last floor, while the potable water quality was analysed in three cold water samples collected at the inlet from the aqueduct network, at the exit from the autoclave, and at the most remote tap. Legionella pneumophila sg1, Legionella pneumophila sg2-16, and non-pneumophila Legionella species were detected in 26% of the hot water networks, mostly in CB and TB. In these buildings we detected correlations between the presence of Legionella and the total chlorine concentration decrease and/or the increase of the temperature. Cold water resulted free from microbiological hazards, with the exception of Serratia liquefaciens and Enterobacter cloacae isolated at the exit from two different autoclaves. We observed an increase in total microbial counts at 22 °C and 37 °C between the samples collected at the most remote taps compared to the ones collected at the inlet from the aqueduct. The study highlights a condition of potential risk for susceptible categories of population and supports the need for measures of risk assessment and control

The Enigmatic Genetic Landscape of Hereditary Hearing Loss: A Multistep Diagnostic Strategy in the Italian Population

Hearing loss is the most frequent sensorineural disorder, affecting approximately 1:1000 newborns. Hereditary forms (HHL) represent 50–60% of cases, highlighting the relevance of genetic testing in deaf patients. HHL is classified as non-syndromic (NSHL—70% of cases) or syndromic (SHL—30% of cases). In this study, a multistep and integrative approach aimed at identifying the molecular cause of HHL in 102 patients, whose GJB2 analysis already showed a negative result, is described. In NSHL patients, multiplex ligation probe amplification and long-range PCR analyses of the STRC gene solved 13 cases, while whole exome sequencing (WES) identified the genetic diagnosis in 26 additional ones, with a total detection rate of 47.6%. Concerning SHL, WES detected the molecular cause in 55% of cases. Peculiar findings are represented by the identification of four subjects displaying a dual molecular diagnosis and eight affected by non-syndromic mimics, five of them presenting Usher syndrome type 2. Overall, this study provides a detailed characterisation of the genetic causes of HHL in the Italian population. Furthermore, we highlighted the frequency of Usher syndrome type 2 carriers in the Italian population to pave the way for a more effective implementation of diagnostic and follow-up strategies for this disease

Application of hydrogen peroxide as an innovative method of treatment for Legionella control in a hospitalwater network

Objectives: To evaluate the effectiveness of hydrogen peroxide (HP) use as a disinfectant in the hospital water network for the control of Legionella spp. colonization. Methods: Following the detection of high levels of Legionella contamination in a 136-bed general hospital water network, an HP treatment of the hot water supply (25 mg/L) was adopted. During a period of 34 months, the effectiveness of HP on Legionella colonization was assessed. Legionella was isolated in accordance with ISO-11731 and identification was carried out by sequencing of the mip gene. Results: Before HP treatment, L. pneumophila sg 2-15 was isolated in all sites with a mean count of 9950±8279 cfu/L. After one-month of HP treatment, we observed the disappearance of L. pneumophila 2-15, however other Legionella species previously not seen were found; Legionella pneumophila 1 was isolated in one out of four sampling sites (2000 cfu/L) and other non-pneumophila species were present in all sites (mean load 3000±2887 cfu/L). Starting from September 2013, HP treatment was modified by adding food-grade polyphosphates, and in the following months, we observed a progressive reduction of the mean load of all species (p < 0.05), resulting in substantial disappearance of Legionella colonization. Conclusion: Hydrogen peroxide demonstrated good efficacy in controlling Legionella. Although in the initial phases of treatment it appeared unable to eliminate all Legionella species, by maintaining HP levels at 25 mg/L and adding food-grade polyphosphates, a progressive and complete control of colonization was obtained

The Italian Draft Law on the \u2018Provisions Concerning the Safeguarding of the Intangible Cultural Heritage\u2019

Intangible cultural heritage in Italy is still in need of a unified approach, capable of providing reliable criteria for identifying its assets and for indicating timescales and means by which they should be safeguarded. In the continued absence of up-to-date, ad hoc state legislation (since the content of those laws which do implement international Conventions is too generic in nature to be sufficiently effective), the Regions have proceeded to act in a somewhat scattered manner, giving rise to an extremely fragmented and very disorderly regulatory framework. The draft law N. 4486, "Provisions Concerning the Safeguarding of the Intangible Cultural Heritage", presented on 12th May 2017 at the Chamber of Deputies of the Italian Republic - as the result of the work of an interdisciplinary and inter-university research team coordinated by Marco Giampieretti, who has drafted the final text with the collaboration of Simona Pinton - seeks to fill the serious void that exists in Italian legal system by aligning it to the principles of international and European law, by redirecting the relevant State and Regional legislation, and by satisfying the fundamental requirements of the national community

Affidabilità di SRAM e microprocessori in ambiente spaziale

In tale elaborato, dopo una breve introduzione sull’architettura dei microprocessori e dei loro principali componenti, verrà dato particolare rilievo ai meccanismi che si instaurano nei circuiti integrati, in particolare nei transistor MOSFET, in seguito all’impatto con le radiazioni. Successivamente verranno proposte numerose tecniche di hardening sia riguardanti l’utilizzo di un particolare processo di fabbricazione sia focalizzate su una riorganizzazione del layout. Tali tecniche riguarderanno sia i singoli transistor, sia i componenti base che si ritrovano all’interno dei microprocessori come ad esempio le memorie SRAM e i registr

Where Integrated Electronics Meets Bio/Micro-Technology: From Synthetic Microstructures to Living Cells On-Chip

The always more predominant synergy between the biological and engineering worlds, is leading to an extremely fruitful mutual cooperation in which biology can benefit of engineering innovations and, vice versa. In this framework, unknown biological processes and complex bio-chemical dynamics can finally be unveiled, tremendously contributing to improve the current health-care and diagnostic paradigm. To reach this ambitious aim, it is fundamental to understand how these two worlds, apparently so distant, can efficiently and mutually integrate. My Ph.D. thesis was focused, specifically, on the interaction between electronics and cardiac cells, due to the intrinsic electrical activity of the latter. Due to the micrometer dimensional scale of the cells, it is advantageous to also use micrometer-scale electrodes, capable to interface with the living entities at their natural and functional dimensions where the most relevant processes happen. For this reason, I analyzed in depth also the use of electrical platforms, embedding micro-electrodes, both passive or with CMOS circuits directly underneath. Due to the complex nature of the problem of the cells viability on-chip, at first synthetic microstructures were used to test and optimize the sensing capabilities of the developed platforms. Specifically, zinc oxide (ZnO) micro-wires were at first deposited across the nanometric-spaced gap induced by exploiting the electromigration phenomenon on a micrometric gold metal wire. Thanks to the physical/electrical properties of ZnO microwires, the main goal was to create a multi-sensor capable to simultaneously detect variations of pH, temperature and UV-visible light, all important parameters to monitor in cell cultures. To improve the robustness of the system against electrical noise, the same microstructures were deposited across couples of micrometer electrodes, but this time fabricated on the surface of a CMOS chip. In this way, the electronic read-out of the variation of the electrical properties of the microstructure, according to the variation of the external parameters selected, was performed directly underneath the electrodes surface, hence reducing noise and parasitics. Specifically, 24 couples of aluminum electrodes, were designed so to allow multiple measurements on the same chip. To improve the quality of the electrical contact between the microstructure and the aluminum electrode, easily oxidizing, a tailored electroless gold plating process was optimized to modify the electrodes surface to gold. The CMOS chip with deposited ZnO microwires was then used to investigate the UV-visible light sensing capabilities of the microstructure. To understand the complex and multiple requirements of performing, instead, measurements of living cardiac cells on chip, a visiting period of 6 months was spent at the Kademhosseini laboratory in Boston (USA), part of the Harvard-MIT health science and technology division. During this time, the needs of primary neonatal rats cardiomyocytes were investigated by developing and electrically modulated bio-hybrid actuator, triggered by the beating activity of cardiac cells, and fabricated using bio-compatible hydrogels. After this extremely important intermediate step, preliminary experiments were done, at the end of the Ph.D. activity by measuring the electrical signals of cardiac cells directly on the surface of CMOS multi-electrodes arrays platforms. Some tests were performed also functionalizing the surface of the CMOS chips with the hydrogel developed in Boston for the bio-hybrid actuator. This activity was performed at the Italian Institute of Technology of Genova (Italy). The main focus of this activity was trying to evoke the firing of an electrical cardiac signal, commonly known as cardiac action potential, by applying an electrical stimulation. At the same time, the biologically-produced electrical signals were also recorded in real-time. Although the performed activity represented only an initial proof-of-concept, it provided an important insight to the challenges to be faced when living entities are deposited on electronic chips. At the same time, the preliminary experiments revealed the huge potentiality of active micro-electronic platforms for the monitoring, study and excitation of living cardiac cells. Thanks to these extremely multi-disciplinary studies it will be possible to significantly impact the current health-care and drug-development paradigms due to the possibility to investigate, previously unknown or not well characterized, biological processes, directly at their proper dimensional scale, with state-of-the-art integrated and hybrid electronic platforms