Serological and molecular identification of Legionella spp. isolated from water and surrounding air samples in Italian healthcare facilities.

BACKGROUND: Legionella is an intracellular microorganism living in natural and artificial aquatic environments. Although its transmission to humans is linked to the inhalation of contaminated aerosols, there is no validated air sampling method for the control and prevention of the disease. The aim of the present study was to provide more information on the distribution of Legionella spp. in indoor environments and to determine whether the same Legionella strains are isolated from air and water samples. METHODS: Ten healthcare facilities located in seven regions of Italy were enrolled. The serological typing of Legionella spp. from water samples and the surrounding air by active and passive sampling was assessed using polyvalent and monovalent antisera. Subsequently, the strains identified as Legionella pneumophila (Lpn) underwent molecular typing by sequence-based typing (SBT) using seven genes (flaA, pilE, asd, mip, mompS, proA, and neuA). The allelic profile number was assigned using the European Working Group for Legionella Infections-SBT database. RESULTS: Lpn serogroup 6 was the most prevalent serogroup; it was found simultaneously in the air and water samples of three different healthcare facilities. In the remaining seven hospitals, Lpn serogroups 1, 6, 7, 9, and 12 were isolated exclusively from water samples. The molecular investigation showed that Lpn strains in the water and air samples of each positive healthcare facility had the same allelic profile. Strains, identified as sequence types (STs) 728 and ST 1638+ST 1324, were isolated in two respective healthcare facilities, and a new strain, identified as ST 1989, was obtained in one healthcare facility. CONCLUSION: The application of the SBT method allowed to verify the homology among Legionella strains from water samples and the surrounding air. The results showed that the same Lpn strains were present in the air and water samples, and a new Legionella strain was identified.

[Legionella spp. contamination in indoor air: preliminary results of an Italian multicenter study]. / La contaminazione indoor da Legionella spp: risultati preliminari di una indagine multicentrica italiana.

OBJECTIVE: To propose a standardized protocol for the evaluation of Legionella contamination in air. DESIGN: A bathroom having a Legionella contamination in water >1,000 cfu/l was selected in 10 different healthcare facilities. Air contamination was assessed by active (Surface Air System, SAS) and passive (Index of Microbial Air, IMA) sampling for 8 hours, about 1 m away from the floor and 50 cm from the tap water. Two hundred liters of air were sampled by SAS every 12 min, after flushing water for 2 min. The IMA value was calculated as the mean value of colony forming units/16 plates exposed during sampling (2 plates/hour). Water contamination was evaluated at T0, after 4 and 8 hours, according to the standard methods. RESULTS: Air contamination by Legionella was found in three healthcare facilities (one with active and two with passive sampling), showing a concomitant tap water contamination (median=40,000; range 1,100-43,000 cfu/l). The remaining seven hospitals isolated Legionella spp. exclusively from water samples (median=8,000; range 1,200-70,000 cfu/l). CONCLUSIONS: Our data suggest that environmental Legionella contamination cannot be assessed only through the air sampling, even in the presence of an important water contamination.

The Presence of Legionella in Water Used for Car Washing: Implications for Public Health.

Although today all of the aspects of Legionella are better understood than in the past, in many countries the interest is still mainly focused on healthcare and tourism facilities. Other at-risk areas are less explored, such as those where workers are often in contact with water during their activities. In reality, any water system capable of producing aerosols can be considered a potential source of Legionella transmission, including car washes, where a large number of users work and flow through annually. From January to May 2022, 120 samples were carried out in 30 car washes located in Messina (Italy): 60 samples of water and 60 of aerosols. The aim of this investigation was to evaluate the risk of legionellosis in car washing workers exposed to potentially contaminated aerosols. To increase the probability of finding Legionella, the sample collections were organized on different days of the week. Of the total samples taken, 10 (8.3%) were positive for Legionella: seven (11.7%) water (range 100-1000 CFU) and three (5%) aerosol (range 10-150 CFU) samples. Detected serogroups were L. pneumophila sgr 1, 7, 10 and Legionella gormanii. Given the results obtained, preventative measures should be implemented in such facilities in order to protect the health of users and car wash operators.

Phenolic Substances in Foods and Anticarcinogenic Properties: A Public Health Perspective.

The interest in polyphenols from vegetable sources has been progressively increased because of the demonstrated correlation between their abundance in certain foods or food preparations of traditional importance and heritage, and the answer of anti-inflammatory strategies in hospitalized patients in the presence of polypehnol-rich foods (as a complementary therapy). Consequently, research involving the accessory role of polyphenols as anti-tumoral aids have been carried out with the aim of finding new additional strategies. The purpose of this paper to evaluate the role of phenolic compounds in foods with reference to health effects for human beings. The importance of these molecules has been evaluated by the health and safety perspectives in terms of: fight to cardiovascular diseases; prevention of chronic-degenerative disorders; general antioxidant properties; and anticarcinogenic features. Moreover, the role of polyphenols-rich foods as anticancer agents has been discussed with relation to two distinct "action plans" on the public hygiene level: the promotion of human health on the one side (for non-hospitalized and normal subjects), and reliable contrasting strategies in cancer patients.

[Energy sources. Social and environmental impact and societal models: future perspectives. Part 1: General aspects and reflections.]. / Fonti energetiche. Impatto socio ambientale e modelli di società: quali prospettive per il futuro.

In this paper, the authors discuss energy sources, highlighting their impact on the environment and on human beings, their influence in economy and finance and on relations between governments. They attempt to analyse whether the above factors together can lead to a negative impact on health, defined as an individual's "complete physical social and psychological well being". The role of petroleum in the world economy is understandable if one considers that energy, heat, light, electricity, transportation and large part of mass production are all dependent on this energy resource. From petroleum one obtains fuel, fertilizers, pesticides, plastic, pharmaceutical products and clothing. Petroleum has become increasingly important in conjunction with expanding globalization and consumerism and the continuous growth of demand for petroleum has led to a corresponding decrease in its production and availability and an increase in its cost, all factors which have led to strong tensions between world States. The authors discuss sea and air pollution and global warming, citing some of the most relevant climatic incidents of recent years and tracing the most important events regarding attempts to contain pollution. They highlight the impact of contaminants such as greenhou se gases, electromagnetic pollution, synthetic chemicals, domestic, industrial and electronic waste products, responsible, according to neo-Lamarckian evolutionists, for the increasing incidence of chronic degenerative diseases. In conclusion the authors stress that there is a need to pursue energy efficiency while awaiting that world States succeed in their common objective of adopting new energy policies, with the use of clean energy at low cost.

[Adolescents and alcohol: a survey in the city of Messina (Italy)]. / Gli adolescenti e l'alcol: indagine statistica nella città di Messina.

Alcohol abuse represents a relevant social problem in industrialized countries. Italy has an age-long tradition of producing and drinking wine, a beverage which in moderate doses is considered to be part of the much appreciated "Mediterranean diet". However, drinking other alcoholic beverages is becoming increasingly common, especially among adolescents. In order to evaluate aspects of the "alcohol" phenomenon among adolescents in the city of Messina (Italy), an anonymous questionnaire was administered during school hours to 5,168 students aged 13 to 17 years. Results show that 989 (19.1%) students, of which 38.3% were females and 61.7% males, reported drinking alcohol. Beer was the preferred drink, followed by alcohol-containing aperitifs, spirits and wine. It was found that students begin drinking alcoholic beverages more regularly at 16 years of age and do so mainly during group encounters. Almost none of the interviewed students reported ever having been drunk, and for most students alcohol is considered to be dangerous only if drunk in excessive amounts (70.9% of females and 72.7% of males). Family habits were found to clearly influence students' degree of alcohol consumption. Information and education is needed by young adolescents regarding the responsible approach to alcoholic beverages.

Environmental Surveillance of Legionellosis within an Italian University Hospital-Results of 15 Years of Analysis.

Legionnaires' disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large and old buildings, such as hospitals, can be contaminated with legionellae and therefore represent a potential risk for the hospital population. In this study, we demonstrated the constant presence of Legionella in water samples from the water system of a large university hospital in Messina (Sicily, Italy) consisting of 11 separate pavilions during a period of 15 years (2004⁻2018). In total, 1346 hot water samples were collected between January 2004 and December 2018. During this period, to recover Legionella spp. from water samples, the standard procedures reported by the 2000 Italian Guidelines were adopted; from May 2015 to 2018 Italian Guidelines revised in 2015 (ISS, 2015) were used. Most water samples (72%) were positive to L. pneumophila serogroups 2⁻14, whereas L. pneumophila serogroup 1 accounted for 18% and non-Legionellapneumophila spp. Accounted for 15%. Most of the positive samples were found in the buildings where the following critical wards are situated: (Intensive Care Unit) ICU, Neurosurgery, Surgeries, Pneumology, and Neonatal Intensive Unit Care. This study highlights the importance of the continuous monitoring of hospital water samples to prevent the potential risk of nosocomial legionellosis.

The contribution of professors of Hygiene to the progress and development of public health in Italy: one hundred years of history.

In 1917 Achille Sclavo, a distinguished researcher and founder of the Italian Society of Hygiene, ended up the first term as Rector of the University of Siena. Since then, the contribution of professors of hygiene in Italian universities has ranged over several relevant topics including vaccinations, environmental hygiene, hospital hygiene, healthcare organization and management, with an important contribution to the health reform of 1978 by Augusto Giovanardi and Alessandro Seppilli. Several Academic Schools (Roman, Neapolitan, Genoese, Sicilian, Venetian, Lombard, etc.) have produced excellent researchers, teachers and mentors who have also occupied important positions in the panorama of the Italian health system. This note analyzes the main research topics, the most famous institutes and departments of hygiene and public health and the contributions of the most famous professors for the development of the discipline, the management of the post- graduated Schools in hygiene and preventive medicine, the commitment in scientific associations and the role of some of them in important institutional positions. Even through its contribution and constant commitment to the institutions, Italian public health has achieved the reputation of being one of the best known and appreciated in the international scientific community.

Phenolic Substances in Foods: Health Effects as Anti-Inflammatory and Antimicrobial Agents.

Background: The interest in phenolic compounds present in foods of vegetable origin has shown a notable increase in recent decades. This interest is due to the growing number of scientific studies concerning their beneficial role in human health. The interest in polyphenols has been supported by the current and growing awareness, and attention of consumers to food from a food safety viewpoint and also because of the beneficial effects ascribed to polyphenols. Objective: The aim of this article is to highlight antibacterial, antifungal, and anti-inflammatory activities of various phenolic compounds normally found in certain foods. Conclusions: Phenolic compounds exert different biological functions, such as antioxidant activity, modulation of detoxifying enzymes, stimulation of the immune system, reduction of platelet aggregation, modulation of hormonal metabolism, reduction of blood pressure, and anti-inflammatory, antibacterial, antiviral, and antifungal activities.

[Prevention of legionellosis in hotel establishments: a proposal to introduce a plan of action in accordance with Provision 13 January 2005 (Italy)]. / La prevenzione della Legionellosi nelle strutture alberghiere. Proposta di un Piano di Autocontrollo ai sensi del Provvedimento 13 Gennaio 2005.

Provision 13 January 2005 (Italy) gives guidelines for legionellosis control and prevention in hotel and thermal establishments. Considering that Legionella pneumophila is an ubiquitous organism , the implementation of actions to control the risk of Legionella in the water distribution systems of various types of establishments (hospital, hotel, thermal), or to at least reduce it to acceptable levels, is warranted. The authors have prepared a plan of action for Legionella which, besides describing the main points of the provision and the roles of managers and technicians, also includes the data sheets to be used during environmental monitoring procedures. The action plan will be freely available through local health authorities and professional associations.

Bacterial isolates from the Arctic region (Pasvik River, Norway): assessment of biofilm production and antibiotic susceptibility profiles.

Bacterial biofilm production is recognized as a strategy that helps aquatic bacteria in resisting to the presence of several kinds of pollutants, including antibiotics, in the bulk environment. The Pasvik River, located between Norway, Russia and Finland, is a sub-Arctic site polluted by wastes from metallurgic and mining activities. In order to study whether and to what extent bacteria are able to produce biofilms, and to assess whether this physiological characteristic influences their resistance to antibiotics, an investigation was performed on bacteria isolated from water and sediment collected along the Pasvik River course during two surveys (May and July). Bacterial strains were screened for their biofilm production and profiles of susceptibility to antibiotics. Results showed that biofilm formation was a widespread characteristic of the isolates. Most of them were also resistant to several antibiotics, such as ampicillin (100% of the isolates) as well as cefazolin, cefoxitin, ceftriaxone, mezlocillin, nitrofurantoin and sisomicin (90% of the total strains). This study shows a significant association between biofilm formation and antibiotic resistance at inner stations both in water and in sediments in May only. This suggests that in Pasvik River colder temperature may stimulate bacterial aggregation into biofilm and simultaneously decrease bacterial susceptibility to antibiotics; since the occurrence of antibiotic resistance has frequently been linked to the presence of pollutants, this result could represent a strategy of bacterial survival under altered environmental conditions.

Legionella risk assessment in cruise ships and ferries.

Introduction. The increasing development of marine traffic has led to a rise in the incidence of legionellosis among travellers. It occurs in similar environments, especially closed and crowded, and aboard ships Legionella survives and multiplies easily in water pipes, spreading into the environment through air conditioning systems and water distribution points. Although in recent years in the construction of cruise ships preventive measures aimed at curbing the proliferation of Legionella (design, materials, focus on the operation and maintenance of the water system), have been taken account, little or no attention has been paid to small ships which, in many cases, are old and not well maintained. Objective. The aim of the study was to evaluate the frequency and severity of Legionella contamination in ferries and cruise ships in order to adopt more specific control measures. Materials and method. A prevalence study was carried out on 10 ferries and 6 cruise ships docking or in transit across the port of Messina (Sicily, Italy). Water and air samples collected from many critical points were tested for qualitative and quantitative identification of Legionella. Results and conclusions. Legionella pneumophila sg 1 was isolated from the samples of shower and tap water in 7 (70%) of the 10 ferries examined, and in 3 (33%) of the 6 cruise ships examined, and L. pneumophila sg 2-14 in 8 (80%) and 1 (16.7%) of these ships, respectively. No Legionella contamination was found in whirlpool baths, air and ice samples. In conclusion, the data obtained confirm higher levels of Legionella contamination in local ferries and cruise ships, underlining the need to adopt corrective actions more specific for these smaller vessels.

Innate Immune Surveillance in the Central Nervous System Following Legionella pneumophila Infection.

BACKGROUND & OBJECTIVE: The innate immune response is a common occurrence in many neuroinflammatory diseases. Central Nervous System (CNS) resident immune cells are able to detect and react to infections and sterile trauma. Peripheral immune cell migration into CNS is regulated by the blood-brain barrier, although peripheral immune cells can invade CNS through meninges, choroid plexus, perivascular spaces, and cerebrospinal fluid. Consequently, in the brain, immune reactions can be mediated by both resident and peripheral immune cells. Both in the periphery and within the CNS, innate immune response is regulated by a wide array of pattern recognition receptors, including Tolllike, scavenger, Retinoic Acid-inducible Gene-1 like, and nucleotide-binding oligomerization domainslike responsible for inflammasome formation. Inflammasome pathway activation induces pyroptosis, a highly inflammatory cell death pattern that occurs to remove intracellular pathogens. Legionella pneumophila is an intracellular microorganism responsible for Legionnaires' disease, a lung infection always associated to neurological dysfunctions. Recent studies have been shown that Toll-like receptors, nucleotide-binding oligomerization domains-like receptors, and RIG-1 like, are activated by L. pneumophila. This flagellated bacterium is able to replicate in phagocytic cells, including macrophages and microglia, responding by activating inflammasome pathways that may be the cause of CNS dysfunction detected in several infected patients. CONCLUSION: The aim of this review is to bring together the latest findings concerning L. pneumophila infection and innate immune host cell responses. A deeper knowledge of these processes could allow the use of immunomodulatory compounds able to counteract CNS involvement following L. pneumophila infection.

Evaluation of Legionella Air Contamination in Healthcare Facilities by Different Sampling Methods: An Italian Multicenter Study.

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®µ) 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®µ 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®µ did not yield Legionella in any enrolled HF. However, molecular investigation using Coriolis®µ 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.

Geostatistics - a tool applied to the distribution of Legionella pneumophila in a hospital water system.

INTRODUCTION: Legionnaires' disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large buildings, such as hospitals, are often contaminated with legionellae and therefore represent a potential risk for the hospital population. The aim of this study was to evaluate the potential contamination of Legionella pneumophila (LP) in a large hospital in Italy through georeferential statistical analysis to assess the possible sources of dispersion and, consequently, the risk of exposure for both health care staff and patients. MATERIALS AND METHOD: LP serogroups 1 and 2-14 distribution was considered in the wards housed on two consecutive floors of the hospital building. On the basis of information provided by 53 bacteriological analysis, a 'random' grid of points was chosen and spatial geostatistics or FAIk Kriging was applied and compared with the results of classical statistical analysis. RESULTS: Over 50% of the examined samples were positive for Legionella pneumophila. LP 1 was isolated in 69% of samples from the ground floor and in 60% of sample from the first floor; LP 2-14 in 36% of sample from the ground floor and 24% from the first. The iso-estimation maps show clearly the most contaminated pipe and the difference in the diffusion of the different L. pneumophila serogroups. CONCLUSION: Experimental work has demonstrated that geostatistical methods applied to the microbiological analysis of water matrices allows a better modeling of the phenomenon under study, a greater potential for risk management and a greater choice of methods of prevention and environmental recovery to be put in place with respect to the classical statistical analysis.

Legionella spp., amoebae and not-fermenting Gram negative bacteria in an Italian university hospital water system.

INTRODUCTION: In hospital and other health care facilities, contamination of water systems by potentially infectious microorganisms, such as bacteria, viruses and protozoa, is a source of nosocomial infections, which may originate fromcolonization of water pipes, cooling towers, spa pools, taps, showers and water supplies. Objective. The study focuses on the occurrence of Legionella spp., free-living amoebae and non-fermenting Gram-negative microorganisms in a University hospital water system located in the town of Messina (Sicily, Italy), which had never been examined previously. MATERIALS AND METHODS: From January 2008 - March 2009, hot tap water samples were collected from 10 wards. Legionella spp. recovered on selective culture medium were identified by microagglutination latex test; free-living amoebae were cultured using Escherichia coli as a food source. Non-fermenting Gram negative microorganisms were identified by API 20 NE strips. RESULTS: Legionella spp. were found in 33.33% of the samples. L. pneumophila serogroup 1 was recovered from the Laboratory Diagnostic and Anaesthesia-Neurology Wards, with a peak of 3.5 × 10(4) cfu/L in May 2008. L. pneumophila serogroups 2-14 were found in the Othorhinolaryngology, Pathologic Anatomy, Paediatrics and Surgery Wards, and peaked (4 × 10(4) cfu/L) in April 2008. Pseudomonadaceae and Hyphomycetes were also detected. Legionella spp. were recovered from samples positive for non-pathogenic amoebae Hartmannella spp. CONCLUSION: This first study of a Messina hospital water system suggested potential health risks related to the detection of Hartmannella spp., as reservoirs for Legionella spp., and Pseudomonas aeruginosa, a Gram negative non-fermenting bacterium frequently causing nosocomial pneumonia. The urgent need for monitoring programmes and prevention measures to ensure hospital water safety is stressed.