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Severe COVID-19 and MIS-C are rare but serious outcomes associated with SARS-CoV-2 infection. The onset of MIS-C often involves the gastrointestinal system, suggesting a potential connection with gut microbiota. This study aims to compare the gut microbiota of children with severe COVID-19 and those with MIS-C using various biomolecular approaches. Gut microbiota composition and specific microbial modulations were analyzed using fecal samples collected at hospital admission. The study included hospitalized patients (mean age 6 ± 5 years) diagnosed with severe COVID-19 (37 patients) or MIS-C (37 patients). Microbial differences were assessed using both NGS and qRT-PCR methodologies. In 75% of cases, pharmacological treatments included antibiotics and corticosteroids, which influenced the microbiota composition. Early age was found to have the most significant impact on microbiota diversity. Significant differences in alpha and beta diversity were observed between COVID-19 and MIS-C patients, particularly concerning low-abundance species. Levels of Bacteroides spp., Bifidobacterium spp., and Akkermansia muciniphila were comparable between groups, while an increased activity of Bifidobacterium spp. was noted in children with positive fecal samples ( p = 0.019). An in-depth evaluation of lesser-known gut species may be key to reducing the risk of severe outcomes and developing microbiota-based biomarkers for the early diagnosis of MIS-C.

Antimicrobial resistance (AMR) is recognized as a global threat. AMR bacteria accumulate in sewage sludge however, knowledge on the persistence of human pathogens and AMR in the sludge line of the wastewater treatment is limited. Sludge can be used, with or without additional treatment, as fertilizer in agricultural fields. The aim of this study is to obtain knowledge about presence of human pathogens and AMR in the sewage sludge, before and after the anaerobic digestion (AD) applying innovative combinations of methods. Fifty sludge samples were collected. Cultivation methods combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Antibiotic Susceptibility Test (AST) were used obtaining knowledge about the microbial community, pathogens, and antibiotic resistant bacteria while the droplet digital Polymerase Chain Reaction (ddPCR) was performed to detect most common AMR genes. In total, 231 different bacterial species were identified in the samples. The most abundant species were spore-forming facultative anaerobic bacteria belonging to Bacillus and Clostridium genera. The AD causes a shift in the microbial composition of the sludge (p = 0.04). Seven pathogenic bacterial species constituting 188 colonies were isolated and tested for susceptibility to Clindamycin, Meropenem, Norfloxacin, Penicillin G, and Tigecycline. Of the Clostridium perfringens and Bacillus cereus isolates 67 and 50 %, respectively, were resistant to Clindamycin. Two B. cereus and two C. perfringens isolates were also resistant to other antibiotics showing multidrug resistance. ARGs (bla OXA , bla TEM , ermB, qnrB, tet(A)-(W), sulI-II) were present at 7-8 Log gene copies/kg of sludge. AD is the main driver of a reduction of some ARGs (1 Log) but resistant bacteria were still present. The results showed the usefulness of the integration of the proposed analytical methods and suggest a decrease in the risk of presence of cultivable pathogens including resistant isolates after AD but a persistent risk of ARGs' horizontal transmission., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Urban air pollution is recognized as a critical problem for public health and is classified as a carcinogen for humans. A great number of studies have focused on the monitoring of urban air mutagenicity. One of the best-known and applied methods for assessing mutagenicity is the Ames test, a bacterial reverse mutation test. The classic protocol for assessing air mutagenicity involves the concentration of particulate matter (PM) on filters and subsequent extraction using organic solvents. This work aimed to develop a method for the evaluation of air mutagenicity directly impacted by air on microbial plates already containing an Ames' microbial sensor., Methods: A specific six-month sampling campaign was carried out in Turin in a period with high air pollution. Samples were tested for mutagenicity on Salmonella typhimurium strains TA98, TA100, and YG1024 with the traditional method and with the new direct method., Results: The new protocol is able to evaluate the mutagenicity of the sampled air and obtain repeatable results. The final sensitivity is similar to the traditional method (≈10 net revertants/m 3 ); however, the mutagenic response is due to the complete air pollution mixture, including volatile and semivolatile pollutants avoiding the concentration of filters and the following laborious extraction procedures., Conclusions: Despite some critical issues in contamination control, the method is easier, faster, and less expensive than traditional methods.

Introduction: Type 1 diabetes (T1D) risk involves genetic susceptibility but also epigenetics, environment, and behaviors. Appropriate metabolic control, especially quickly after the diagnosis, is crucial for the patient quality of life., Methods: This study aimed to produce a quantitative comparison of the behavior, nutrition habits, and gut microbiota composition between the onset and the 1-year follow-up in 35 children with T1D., Results and Discussion: At follow-up, with the metabolic control, many parameters improved significantly, with respect to the onset, such as glycated hemoglobin (-19%), body mass index (BMI), and also nutritional behaviors, such as normal calorie intake (+6%), carbohydrate intake (-12%), extra portion request (-4%), and meals distribution during the day. Moreover, glycated hemoglobin decrement correlated with both total and rapid absorption carbohydrate intake (Spearman's rho = 0.288, 95% CI 0.066-0.510, p = 0.013), showing as the nutritional behavior supported the insulin therapy efficiency. The next-generation sequencing (NGS) analysis of microbiota revealed abundance differences for Ruminococcus bromii and Prevotella copri (higher at onset, p < 0.001) and the genera Succinivibrio and Faecalibacterium (lower at onset, p < 0.001), as a consequence of nutritional behavior, but it was not the only changing driver. The qRT-PCR analysis showed significant variations, in particular for Bacteroidetes and Bifidobacterium spp. (+1.56 log gene copies/g stool at follow-up, p < 0.001). During the year, in 11% of the patients, severe clinical episodes occurred (hypoglycemic or ketoacidosis). The likelihood of a severe hypoglycemic episode was modulated when the Methanobrevibacter smithii amount increased (odds ratio 3.7, 95% CI 1.2-11.4, p = 0.026). Integrated evaluation, including nutritional behavior and microbiota composition, could be considered predictive of the metabolic control management for children cohort with a recent diagnosis of T1D., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Traversi, Scaioli, Rabbone, Carletto, Ferro, Franchitti, Carrera, Savastio, Cadario, Siliquini, Cerutti and Durazzo.)

Environmental pollutants can influence microbiota variety, with important implications for the general wellbeing of organisms. In subjects at high-risk of cancer, gut, and lung microbiota are distinct from those of low-risk subjects, and disease progression is associated with microbiota alterations. As with many inflammatory diseases, it is the combination of specific host and environmental factors in certain individuals that provokes disease outcomes. The microbiota metabolites influence activity of epigenetic enzymes. The knowledge of the mechanisms of action of environmental pollution now includes not only the alteration of the gut microbiota but also the interaction between different human microbiota niches such as the lung-gut axis. The epigenetic regulations can reprogram differentiated cells in response to environmental changes. The microbiota can play a major role in the progression and suppression of several epigenetic diseases. Accordingly, the maintenance of a balanced microbiota by monitoring the environmental stimuli provides a novel preventive approach for disease prevention. Metagenomics technologies can be utilized to establish new mitigation approaches for diseases induced by polluted environments. The purpose of this review is to examine the effects of particulate matter exposure on the progression of disease outcomes as related to the alterations of gut and lung microbial communities and consequent epigenetic modifications.

The finest fraction of Particulate Matter (PM2.5) carries a large number of pollutants, some of which are assessed as genotoxic, such as some Polycyclic Aromatic Hydrocarbons (PAHs). In many countries, PM2.5 in combination with some PAHs are monitored to assess the concentrations of pollutants, while the air quality is rarely assessed by means of biological assays. Epidemiological studies have demonstrated a significant correlation between these two pollutants and human adverse effects, in particular on the respiratory system. Nevertheless, other air pollutants can induce a biological effect and the cumulative effect of the PM2.5 complex mixture may not be easily deduced by PM2.5 and PAH levels. This study aimed to combine the legislative monitoring of PM2.5 with the study of its mutagenicity. During a full year, daily air samples were collected in nine sites of the North-western Italy air quality monitoring network (Piedmont Region) and PM2.5 and PAH concentrations were assessed. Monthly pooled organic extracts were tested with the Salmonella assay using TA98 and TA100 strains, with and without metabolic activation (±S9), and using TA98NR and YG1021 strains. In all sites, a positive response was observed for TA98 and TA100 especially without S9. A significant mutagenic seasonal variation was detected, with higher mutagenicity in winter and lower responses in summer (average total mutagenicity ratio 27:1). The response of TA98NR and YG1021 compared with TA98 suggested a significant contribution of nitro-compounds to the mutagenicity. No significant differences were found between urban background and rural sites denoting the spread of pollution. A mutagenicity increase, 1.28 Total Mutagenicity Factor/20 m 3 , was observed for each PM2.5 μg increment. PAH levels and corresponding Toxic Equivalent Factors were highly correlated to mutagenicity results. This work confirms that complex environmental mixtures can be appropriately assessed through the implementation of physical-chemical analyzes with bioassays able to evaluate synergistic and antagonistic effects, especially for highest and lowest pollution settings., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The development of high-throughput omics technologies represents an unmissable opportunity for evidence-based prevention of adverse effects on human health. However, the applicability and access to multi-omics tests are limited. In Italy, this is due to the rapid increase of knowledge and the high levels of skill and economic investment initially necessary. The fields of human genetics and public health have highlighted the relevance of an implementation strategy at a national level in Italy, including integration in sanitary regulations and governance instruments. In this review, the emerging field of public health genomics is discussed, including the polygenic scores approach, epigenetic modulation, nutrigenomics, and microbiomes implications. Moreover, the Italian state of implementation is presented. The omics sciences have important implications for the prevention of both communicable and noncommunicable diseases, especially because they can be used to assess the health status during the whole course of life. An effective population health gain is possible if omics tools are implemented for each person after a preliminary assessment of effectiveness in the medium to long term.

Background: Multisystem Inflammatory Syndrome in Children (MIS-C) is a rare but severe illness associated with SARS-CoV-2 infection. A dysregulated immune response is recognized as the main pathogenic mechanism. Previous studies demonstrated the presence of SARS-CoV-2 RNA in faeces of almost one-third of patients with COVID-19, while data are currently missing about MIS-C., Objectives: to evaluate faecal sample positivity to SARS-CoV-2 in MIS-C and to compare the positivity rate between MIS-C and COVID-19 hospitalised children.  DESIGN: observational descriptive study with prospective patient enrollment., Setting and Participants: the SARS-CoV-2 positivity was evaluated in stool samples obtained in a prospective series of 63 paediatric patients admitted to Regina Margherita Children's Hospital (Azienda Ospedaliero Universitaria - Città della Salute e della Scienza, Turin, Northern Italy) with diagnosis of MIS-C (N. 31) or COVID-19 (N. 32), during the first year of pandemic emergency. The real-time reverse transcription polymerase chain reaction (real-time RT-PCR), was performed using a validated kit measuring 3 target SARS-CoV-2 genes: E gene, N gene, and ORF1ab gene MAIN OUTCOME MEASURES: SARS-CoV-2 stool positivity and concomitant gastrointestinal symptoms., Results: overall, 16/63 (25%) stool samples revealed the presence of SARS-CoV-2 mRNA. In patients with COVID-19, faecal samples were collected 8 days as median (IQR 7) after the presumed viral exposure and were positive in 12/31 (39%; 95%CI 23.2-56.2); among children with MIS-C, stools were collected 27.5 days as median (IQR 26.25) after presumed contact and the positivity rate was 12.5% (95%CI 4.4-27.0) (4/32). More than 80% of the children with MIS-C presented gastrointestinal symptoms, but the frequency of gastrointestinal symptoms in patients with positive stools for SARS-CoV-2 RNA is not higher than patients tested negative (p=0.092)., Conclusions: MIS-C patients frequently experienced gastrointestinal symptoms, confirming the intestinal involvement in MIS-C already described in the literature. The presence of SARS-CoV-2 mRNA in faecal samples is confirmed in more than 10% of MIS-C patients and stool positivity was also detected many days after presumed first contact with the virus. This data suggests the possibility of tracing SARS-COV-2 also in faeces for a better description of its circulation and spread in the environment.

Aims: The incidence of type 1 diabetes has increased over the last decades. The pathological pathway is not yet clear, even if genetic and environmental risk factors are known. An early diagnosis can avoid ketoacidosis and its complications. This work aims to discuss the determinants of both ketoacidosis at the onset and access by hospital emergency departments without a suspected diagnosis., Methods: An observational bi-centric prospective study was conducted in Northern Italy, on a paediatric population including Italian and migrant patients at the diabetes onset. Seventy-four type 1 diabetes patients, both Italian and migrant, were included in the study. Anthropometric, socio-economic, behavioural, clinical data were collected, and microbiota analyses were performed using stool samples., Results: Regular physical activity is associated with lower ketoacidosis incidence at onset (OR 0.33 95% CI 0.12-0.95 p < 0.05), as is higher blood vitamin D level (OR 0.92 95% CI 0.85-0.99 p < 0.05). Moreover, a higher weaning age (OR 0.49 95% CI 0.27-0.89 p < 0.05), higher vitamin D level (OR 0.90 95% CI 0.83-0.98 p < 0.05) and a higher level of Akkermansia muciniphila (OR 0.46 95% CI 0.25-0.87 p < 0.05) are associated factors to lower frequency of type 1 diabetes onset without a suspected diagnosis. Diabetes migrant status is not a risk factor for severe type 1 diabetes onset; on the other hand, some protective factors are significantly more diffused among Italians, such as regular sport activity and non-critical vitamin D levels., Conclusion: Behavioural and nutritional data, such as microbiota bio-indicators, seem to be useful to identify an at-risk population to prevent ketoacidosis and its severe complications.

Type 1 diabetes (T1D) is a common autoimmune disease that is characterized by insufficient insulin production. The onset of T1D is the result of gene-environment interactions. Sociodemographic and behavioural factors may contribute to T1D, and the gut microbiota is proposed to be a driving factor of T1D. An integrated preventive strategy for T1D is not available at present. This case-control study attempted to estimate the exposure linked to T1D to identify significant risk factors for healthy children. Forty children with T1D and 56 healthy controls were included in this study. Anthropometric, socio-economic, nutritional, behavioural, and clinical data were collected. Faecal bacteria were investigated by molecular methods. The findings showed, in multivariable model, that the risk factors for T1D include higher Firmicutes levels (OR 7.30; IC 2.26-23.54) and higher carbohydrate intake (OR 1.03; IC 1.01-1.05), whereas having a greater amount of Bifidobacterium in the gut (OR 0.13; IC 0.05 - 0.34) was a protective factor for T1D. These findings may facilitate the development of preventive strategies for T1D, such as performing genetic screening, characterizing the gut microbiota, and managing nutritional and social factors.

Aim: To develop an instrument to investigate knowledge and predictive factors of needlestick and sharps injuries (NSIs) in nursing students during clinical placements., Design: Instrument development and cross-sectional study for psychometric testing., Methods: A self-administered instrument including demographic data, injury epidemiology and predictive factors of NSIs was developed between October 2018-January 2019. Content validity was assessed by a panel of experts. The instrument's factor structure and discriminant validity were explored using principal components analysis. The STROBE guidelines were followed., Results: Evidence of content validity was found (S-CVI 0.75; I-CVI 0.50-1.00). A three-factor structure was shown by exploratory factor analysis. Of the 238 participants, 39% had been injured at least once, of which 67.3% in the second year. Higher perceptions of "personal exposure" (4.06, SD 3.78) were reported by third-year students. Higher scores for "perceived benefits" of preventive behaviours (13.6, SD 1.46) were reported by second-year students., Competing Interests: Roger Watson, Editor in Chief, and Mark Hayter, Editor of the Journal of Advanced Nursing, hold honorary positions at the University of Genoa where the study was conducted., (© 2020 The Authors. Nursing Open published by John Wiley & Sons Ltd.)

Urban air pollution represents a global problem, since everyday many mutagenic and carcinogens compounds are emitted into the atmosphere, with consequent adverse health effects on humans and biota. Specifically, particulate matter air pollution was associated with increased risks in human mortality and morbidity. In this paper, we analyse the genomic effects on human lymphocytes of different concentrations of annual Turin PM2.5 extract by an in vitro micronuclei assay. Samplings were collected from an urban meteorological-chemical station positioned in Turin (Italy), one of the most polluted cities in Europe. PM2.5 sampled on filters was used for organic extraction in monthly pools and successively aggregated to produce a mixture representative for a full year PM2.5 collection. Lymphocytes were exposed to four concentrations of PM2.5: 5, 10, 15 and 20 μg/mL and micronuclei, nucleoplasmic bridges and nuclear buds were scored. With respect to controls, PM2.5 significantly increased the frequencies of all analysed biomarkers at all tested concentrations, whereas the CBPI index was significantly reduced only at the concentration of 20 μg/mL. Such in vitro effects can both to stimulate local authorities to adopt efficient measures for air pollution mitigation and to improve human monitoring to detect early precancer lesions.

Background: There are 1.2 million of immigrant children living in Italy. However, data on their nutritional habits are limited. The aim of this study was to assess the nutritional profile in a cohort of both Italian and immigrant children., Methods: The study included 86 children aged 5-15 consecutively enrolled from January 2016 to May 2017 within a larger epidemiological study on determinants of diabetes. Immigrant state was defined on the basis of the parent origin. Data on nutritional profile, frequency of food group consumption, and eating habits were collected using the 24-hour dietary recall method and a questionnaire. Anthropometric parameters were measured., Results: In the cohort of immigrant children there was a higher prevalence of both overweight (27.3 vs. 14.1%) and obesity (18.2 vs. 3.1%) subjects and a greater total calorie intake compared to Italian children, mainly due to excess simple carbohydrate intake. Immigrant children had a higher consumption of sweets, snacks, and drinks with added sugar. Moreover, unhealthy habits, such as eating alone and eating while watching TV, were more frequent among immigrant children., Conclusions: In this cohort, immigrant children had a higher prevalence of overweight/obesity possibly due to less healthy nutritional habits. Culturally-tailored nutritional interventions may help preventing the development of obesity-related diseases in this population.

Bioaerosol exposure linked to the bioenergy production from waste and its effects on human health in occupational and residential environments has rising interest nowadays. The health risk associated with the exposure includes mainly infective diseases, allergies, chronic bronchitis, and obstructive pulmonary disease. A risk assessment's critical point is the bioaerosol quality and quantity characterization. The aim of this study is to evaluate the application of different methods for the analysis of bioaerosol sampled in bioenergy plants. This study involved six Italian plants for the treatment of biomasses and energy production. Bioaerosol cultural evaluation was performed, by Surface Air System (SAS) sampler, and DNA was extracted from PM0.49 samples and Low Melting Agar plates. qRT-PCR followed by Denaturing Gradient Gel Electrophoresis (DGGE) and band sequencings were performed. The cultural method is able to detect less than 15% of what is evaluable with bio-molecular methods. A low sample alfa-diversity and a high beta-biodiversity in relation to feedstock and process were observed. Sequencing showed microorganisms with a hygienic-sanitary relevance such as Arcobacter , Pseudomonas , Enterobacter , Klebsiella , Enterococcus and Bacillus . Integrated cultural and biomolecular methods can be more exhaustive to evaluate bioaerosol's exposure in the occupational environment.

The characterization of anodic microbial communities is of great importance in the study of microbial fuel cells (MFCs). These kinds of devices mainly require a high abundance of anode respiring bacteria (ARB) in the anode chamber for optimal performance. This study evaluated the effect of different enrichments of environmental freshwater sediment samples used as inocula on microbial community structures in MFCs. Two enrichment media were compared: ferric citrate (FeC) enrichment, with the purpose of increasing the ARB percentage, and general enrichment (Gen). The microbial community dynamics were evaluated by polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR-DGGE) and real time polymerase chain reaction (qPCR). The enrichment effect was visible on the microbial community composition both during precultures and in anode MFCs. Both enrichment approaches affected microbial communities. Shannon diversity as well as β-Proteobacteria and γ-Proteobacteria percentages decreased during the enrichment steps, especially for FeC (p < 0.01). Our data suggest that FeC enrichment excessively reduced the diversity of the anode community, rather than promoting the proliferation of ARB, causing a condition that did not produce advantages in terms of system performance.

Anaerobic digestion is a consolidated biotechnology able to produce renewable energy from biomasses. In the European countries, quick growth of biogas production from different organic matrices including wastes has been observed. In relation to the characteristics and quantity of the anaerobic digestion of feedstock, there are different technologies, advantages and criticisms. An accurate occupational risk assessment and development of management tools for green jobs involved in the anaerobic digestion plants are due. The aim of this work is to assess the aerosol exposure for such workers, focusing on the bioaerosol risk. Full scale plants for the treatment of organic municipal waste, waste water treatment sludge, agro zootechnical and food producing byproducts were involved for this purpose. The bioaerosol levels were monitored during activities through culturing and biomolecular methods; moreover, the sub-fractionated PM10 and carried endotoxins were measured in different plant areas. Global microbial contamination is higher (>5000 UFC/m 3 ) in the area where organic wastes are handled and pretreated, both for organic municipal waste plants - with a bacterial prevalence - and agro zootechnical plants - with a fungi prevalence. Moreover, the microbial contamination is higher where organic municipal waste is present in respect to other biomasses (ANOVA p < 0.01). Numerous pathogens are carried by the aerosol. HAdV-4 presence is lower than LOQ (50 gene copies/m 3 ) in all the samples. Environmental PM10 reached the 280 μg/m 3 level including PM3 for 78%. Endotoxin pollution overtakes the 90 EU/m 3 limit sporadically. Personal PM4.5 reached 10 mg/m 3 only for maintenance technicians in the pretreatment area for organic municipal waste. The risk can be evaluated under a quantitative and qualitative point of view highlighting risk management improvement for anaerobic digestion plants., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Wastewater treatment plants (WWTP) are an important source of surface water contamination by enteric pathogens, affecting the role of environmental water as a microbial reservoir. We describe the release to the environment of certain anaerobes of human and environmental concern. The work was focused on emerging microbial targets. They are tracing, by RT-qPCR, on WWTP effluents, both liquid and solid, when an anaerobic digestion step is included. The focus is placed on Clostridium spp. with the specific quantification of Clostridium perfringens, as typical bioindicator, and Clostridium difficile, as emerging pathogen not only confined into nosocomial infection. Moreover methanogens were quantified for their involvement in the anaerobic digestion, and in particular on Methanobrevibacter smithii as major methanogenic component of the human gut microbiome and as not conventional faecal indicator. In the water samples, a reduction, statistically significant, in all microbial targets was observed (p < 0.01), 2 log for the total bacteria, 1.4 log for the Clostridium spp. and M. smithii, 1 log for total methanogens, C. perfringens and C. difficile. The AD process contribute to a significant change in microbial levels into the sludge for total bacteria and total methanogens (p < 0.01), both when the input sludge are primary and secondary, while for the presence of Clostridium spp. and C. difficile there was not a significant change. The produced data are innovative showing which is the diffusion of such anaerobic microorganisms throughout the WWTP and opening a discussion on the implementation of possible techniques for a more efficient microbial removal from effluents, particularly bio-solids, to reduce the potential release of pathogens into the environment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: The anaerobic digestion is one of the most spread renewable energy technology. The input biomasses included various environmental problematic wastes such as sludge coming from wastewater treatment plant (WWTP) and organic fraction of municipal solid waste (OFMSW). As biomolecular procedures have become important tools for the microbial characterisation of anaerobic samples coming from the reactors, it is crucial sampling and extracting properly DNA in order to employ such types of techniques. The current study is aimed to evaluate how freezing temperature and length of storage at -20 °C influence both the extracted DNA yield and microbial community quantifications from digested sludge samples collected at full-scale plants., Results: From WWTP sludge samples, we observed a reduction of DNA concentration comparing fresh and stored samples for 10 days at -20 °C (ANOVA test p < 0.0001), with an estimated DNA loss of approximately 65% for such types of samples, however the methanogen communities can be assessed respecting the fresh conditions. From OFMSW sludge samples, we observed a reduction in extracted DNA (-90%), after 120 frozen days, while microbial communities are determined respecting the fresh conditions within 2 months of frozen storage., Conclusions: The remarkable effect of frozen storage on sludge samples suggests as the better procedure to perform the DNA extraction from fresh sample. On the other hand it is not generally possible, so approximately 2 months of storage at -20 °C appears to be suitable time at which DNA concentrations remain sufficient to perform coherent microbial characterization through quantitative qRT-PCR.

Anaerobic digestion was introduced into wastewater treatment plants several years ago, but anaerobic digestion performance has not yet been achieved. The variability of the microbial community in digesters is poorly understood, and despite the crucial role of anaerobic digestion reactors, the microbial equilibrium that yields the best performance in these reactors has only recently been hypothesised. In this study, two full-scale continuous anaerobic reactors, placed in Torino's main wastewater treatment plant in northern Italy, were followed to develop a summary indicator for measuring anaerobic digestion performance. A total of 100 sludge samples were collected. The samples were characterised chemically and physically, and microbial groups were quantified by qRT-PCR. A chemical biological performance index strictly correlated to specific biogas production (rho=0.739, p<0.01) is proposed. This approach will produce new management tools for anaerobic digestion in wastewater treatment plants., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Commonly the atmospheric pollution research is focussed on particulate indicators especially when mutagenicity was studied. On the other hand the volatile and semi-volatile compounds no adsorbed on to the particles can be genotoxic and mutagenic. Moreover some mutagenic compounds, such as polycyclic aromatic hydrocarbons, are present both in the particulate and in the gas-phase in according to chemical conditions. This work is focussed on the assessing of the total mutagenicity shifting the gas-phase and particulate phase, during two seasons, in Turin. Two sampling sessions are conducted for total particulate matter and gas-phase pollutants. Moreover meteorological and usual air pollution monitoring data were collected at the same sampling station. The Salmonella assay using the strains TA98 and YG1021 was conducted on each organic extract. The mean level of total suspended particles, PM10 and PM2.5 were 73.63±26.94, 42.85±26.75 and 31.55±26.35 μg m(-)(3). The observed mutagenicity was PM induced YG1021>PM induced TA98>PM induced TA98+S9≫non-particle induced YG1021>non-particle induced TA98>non-particle induced TA98+S9. The multivariate regression is significant when we consider air pollution and meteorological indicators and chemical conditions as predictors., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The continued expansion of the green economy increases the risk profile for green occupational jobs. One of the broadest green sectors in terms of growth is the anaerobic digestion of biomasses. In recent years, this development has also interested Italian regions. The management of biomass includes biological risk and the risk of particulate and endotoxin exposure. In the present study, we evaluated airborne exposure for anaerobic digestion workers at two real-scale plants. Digested biomass has different origins, ranging from cattle sludge and manure to poultry manure to agricultural harvesting or processing residues, particularly from maize and fruits. Two sampling points were chosen: at the first, the input biomasses were stored, and the hopper was loaded; at the second, the digested sludge exited the digester. The microbiological parameters, assessed using an active sampler and cultural method, were the total bacteria counts (at 22, 37, and 55°C), yeasts, fungi, Pseudomonaceae, Clostridia spp., Enterobacteriaceae and Actinomycetes. Moreover, at the same sampling points, we evaluated six PM10 fraction levels (10.0-7.2, 7.2-3.0, 3.0-1.5, 1.5-0.95, 0.95-0.49, and <0.49µm) and the endotoxin content of each fraction. In this investigation, the microbe contamination of the air varied from low to high levels, while the PM10 and endotoxin levels were limited, reaching rural environmental levels (61.40µg/m(3) and 18.88EU/m(3), respectively). However, contamination and occupational risk must be evaluated individually for each plant because numerous variables influence the risk magnitude, particularly digested sludge treatments, such as input biomass nature, storage, movement conditions, building configuration and technological processes., (Copyright © 2015 Elsevier Inc. All rights reserved.)

A number of compounds found in particulate matter with an aerodynamic diameter <2.5 (PM2.5) can interact with DNA either directly or after enzymatic transformation to induce DNA modifications. These particulate matter (PM)-induced alterations in DNA may be associated with increased frequencies of pollution-associated diseases, such as lung cancer. In the present study, we applied different methods to assess the mutagenicity and genotoxicity of monthly PM2.5 organic extracts collected over a full year. We used the Salmonella assay, exposed cultured human embryonic lung fibroblasts and applied extracellular lactate dehydrogenase (LDH) and 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assays to assess the cytotoxicity of PM2.5 on the cells. We assessed both the expression levels of a number of DNA repair genes (using qRT-qPCR) and the genetic profile of the treated cells compared to the control. The expression levels of XRCC1 and APE1, which are involved in the first steps of base excision repair, as well as ERCC1, XPA and XPF, which encode nucleotide excision repair subunits, were analysed. The monthly mean of the PM2.5 collected was 35.16 ± 22.06 μg/m(3). The mutagenicity of PM2.5 to TA98 was 46 ± 50 net revertants/m(3), while the mutagenicity to TA98 + S9 was 17 ± 19 net revertants/m(3). The mean IC50 values were 2.741 ± 1.414 and 3.219 ± 2.764 m(3) of equivalent air in the XTT and LDH assays, respectively. A marked and significant increase in APE1 expression levels was observed in the exposed cells. This effect was also significantly correlated with mutagenicity (p < 0.01). No induced AFLP fragment profile alterations were detected. The proposed approach seems to be useful for integrated evaluation and for highlighting the mechanisms inducing DNA damage.

Recently, much attention has been devoted to urban air pollution because epidemiological studies have reported health impacts related to particulate matter (PM). PM10 and PM2.5 were collected during different seasons in Torino, a northern Italian city, and were characterised by inorganic chemical species (secondary particulates and bio-available iron). The biological effects of aqueous and organic solvent PM extracts on human epithelial lung A549 were evaluated, and the effects on cell proliferation and lactate dehydrogenase (LDH) release were assayed. The average PM10 concentration during the sampling period was 47.9 ± 18.0 μg/m(3); the secondary particles accounted for 49 % ± 9 % of the PM10 total mass, and the bio-available iron concentration was 0.067 ± 0.045 μg/m(3). The PM2.5/PM10 ratio in Torino ranged from 0.47 to 0.90 and was higher in cold months than in warm months. The PM10 and PM2.5 extracts inhibited cell proliferation and induced LDH release in a dose-dependent manner with a seasonal trend. The PM10 extract had a stronger effect on LDH release, whereas the PM2.5 extract more strongly inhibited cell proliferation. No significant differences were observed in the effects induced by the two extracts, and no significant correlations were found between the biological effects and the PM components evaluated in this study, thus emphasising the importance of the entire mixture in inducing a cytotoxic response.

The public has recently raised concerns regarding potential human health and environmental risks associated with tire crumb constituents in the artificial turf of football fields. The aim of the present study was to develop an environmental analysis drawing a comparison between artificial turf football fields and urban areas relative to concentrations of particles (PM10 and PM2.5) and related polycyclic aromatic hydrocarbons (PAHs), aromatic hydrocarbons (BTXs), and mutagenicity of organic extracts from PM10 and PM2.5. No significant differences were found between PM10 concentrations at an urban site and on a turf football field, both during warm and in cold seasons, either with or without on-field activity. PM2.5 concentrations were significantly greater at the urban site in the cold season as was the ratio of PM2.5 to PM10. BTXs were significantly greater at urban sites than on turf football fields on both on warm and cold days. The ratio of toluene to benzene (T/B ratio) was always comparable with that of normal urban conditions. The concentration of PAHs on the monitored football fields was comparable with urban levels during the two different sampling periods, and the contribution of PAHs released from the granular material was negligible. PM10 organic extract mutagenicity for artificial turf football fields was greater, whereas PM2.5 organic extract mutagenicity was lower, compared with the urban site studied. However, both organic extract mutagenicity values were comparable with the organic extract mutagenicity reported in the literature for urban sites. On the basis of environmental monitoring, artificial turf football fields present no more exposure risks than the rest of the city.

Biogas is an energy source that is produced via the anaerobic digestion of various organic materials, including waste-water sludge and organic urban wastes. Among the microorganisms involved in digestion, methanogens are the major microbiological group responsible for methane production. To study the microbiological equilibrium in an anaerobic reactor, we detected the methanogen concentration during wet digestion processes fed with pre-treated urban organic waste and waste-water sludge. Two different pre-treatments were used in successive experimental digestions: pressure-extrusion and turbo-mixing. Chemical parameters were collected to describe the process and its production. The method used is based on real-time quantitative PCR (RT-qPCR) with the functional gene mcrA as target. First, we evaluated the validity of the analyses. Next, we applied this method to 50 digestate samples and then we performed a statistical analysis. A positive and significant correlation between the biogas production rate and methanogen abundance was observed (r = 0.579, p < 0.001). This correlation holds both when considering all of the collected data and when the two data sets are separated. The pressure-extrusion pre-treatment allowed to obtain the higher methane amount and also the higher methanogen presence (F = 41.190, p < 0.01). Moreover a higher mean methanogen concentration was observed for production rate above than of 0.6 m(3) biogas/kg TVS (F = 7.053; p < 0.05). The applied method is suitable to describe microbiome into the anaerobic reactor, moreover methanogen concentration may have potential for use as a digestion optimisation tool., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: this article summarises the results of eight years (2002-2009) of investigations on food borne disease outbreaks carried out the Regional Agency for Public Health in Piedmont, which is an Italian Region (4,432,571 citizens, 2008)., Methods: the investigations were conducted as cohort studies supported by laboratory analysis of: food and patients. The data were reported by general practitioners and hospital physicians, and were collected, evaluated and recorded in 22 Hospitals., Results: it recorded 803 outbreaks of food borne disease, of which 177 mushroom outbreaks with 318 ill people and 499 outbreaks for all the others agents with 3,381 ill people. The maximum prevalence was 15.7/100,000 (2004). The most important causes for numbers of ill people have been (2002-2009): salmonellosis (34%, 1,250/3,699), mushroom poisoning (9%, 318/3,699), Escherichia coli (8%, 228/3,699), Staphyilococcus (6%, 209/3,699), virus (5%, 198/3,699), Clostridium perfringens (4%, 154/3,699), Bacillus cereus (2.1%, 76/3,699), scombrotoxin (1.8%, 65/3,699). 303 out of 304 mushrooms outbreaks occurred from consumers that picking mushrooms themselves to eat in private homes and for all others agents: 57% of 499 the outbreaks occurred in private homes, 27% in restaurants, 6% in canteens, but 39% of 3,381 ill people had eaten in restaurants and other public food facilities, 32% in private homes and 24% in canteens. After mushrooms the eggs (75 outbreaks) were the most important food recognized., Conclusion: the theoretical maximum prevalence which can be hypothesized with this data in Piedmont (2002-2009) is 141/100,000 (sum of the maximum prevalence recorded at least in one Piedmont Hospital district).

Methanogen populations and their domains are poorly understood; however, in recent years, research on this topic has emerged. The relevance of this field has also been enhanced by the growing economic interest in methanogen skills, particularly the production of methane from organic substrates. Management attention turned to anaerobic wastes digestion because the volume and environmental impact reductions. Methanogenesis is the biochemically limiting step of the process and the industrially interesting phase because it connects to the amount of biogas production. For this reason, several studies have evaluated the structure of methanogen communities during this process. Currently, it is clear that the methanogen load and diversity depend on the feeding characteristics and the process conditions, but not much data is available. In this study, we apply a Real-Time Polymerase Chain Reaction (RT-PCR) method based on mcrA target to evaluate, by specific probes, some subgroups of methanogens during the mesophilic anaerobic digestion process fed wastewater sludge and organic fraction of the municipal solid waste with two different pre-treatments. The obtained data showed the prevalence of Methanomicrobiales and significantly positive correlation between Methanosarcina and Methanosaetae and the biogas production rate (0.744 p < 0.01 and 0.641 p < 0.05). Methanosarcina detected levels are different during the process after the two pre-treatment of the input materials (T-test p < 0.05). Moreover, a role as diagnostic tool could be suggested in digestion optimisation.

Background: Mass gatherings require attention toward planning and execution of dedicated medical care., Objectives: The aims were to describe the organization and provision of medical care in the Torino 2006 Winter Olympic Games in light of the epidemiology of illnesses and injuries among athletes during this event., Methods: The organization integrated 18 medical centers operating at Olympic venues, three polyclinics inside the Olympic Villages, and 13 pre-existing Olympic Hospitals. The design was a retrospective study, with review of medical records at Olympic medical facilities. All the athletes who attended any medical center or polyclinic were eligible. The main outcomes included type of injuries and illnesses affecting athletes, incidence of injuries and illnesses by provenance and discipline, and rate of hospital admissions., Results: A total of 330 athletes received medical care and 676 medical encounter forms were documented, with an overall medical utilization rate of 12.66%. Skeleton, snowboard, bobsleigh, and alpine and freestyle skiing had the highest medical attendance rate relative to other disciplines (p < 0.001). The majority of injuries and illnesses were of a minor nature. A total of 23 visits required hospital admissions (3.40%)., Conclusion: The described model of medical coverage for such events seemed to be practical and efficient for the athletes, without depleting services to the general population. The risk of severe injury and illness seemed to be low. These data could be useful for planning medical services in future mass gatherings., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Human type II alveolar cells (A549) were exposed to aqueous- and organic-solvent PM10 extracts to evaluate their effects on cell proliferation, proinflammatory cytokine release and cytotoxicity (assayed by lactate dehydrogenase, LDH, activity). PM10 samples collected in Torino (northwest Italy) were analysed for inorganic chemical species (bioavailable iron and secondary particulates) and endotoxins, which are potentially inflammatory promoters in human airways. During the sampling period, PM10 concentration was 55.4±39.1μg/m(3), secondary particles constituted 42±9% of the PM10 total mass, and bioavailable iron concentration was 0.078±0.095μg/m(3). PM10 inhibits cell proliferation and induces both IL-6 and LDH release in a dose- and time-dependent manner, with a seasonal trend. The different effects of aqueous and organic PM10 extracts demonstrate the importance of particle composition for the induction of cytotoxic effects on A549 cells. A first comparison between the biological effects induced by PM10 extracts and PM10 components was performed., (Copyright © 2010 Elsevier B.V. All rights reserved.)

PM2.5 is the breathable fraction of the particulate matter and some adverse health effects, such as respiratory functionality, cardiological diseases and cancer, can be in some measure attributable to this risk factor exposure. Some of the most carcinogen compounds transported by PM2.5 are nitro-compounds. In this study, a strengthened in vitro bioassay--able to predict the mutagenic/carcinogenic activity of the environmental mixtures--was conducted on PM2.5 organic extracts to define the nitro-compounds burden. PM2.5 air pollution was daily monitored, during 2006, in three cities located in the Northern part of Italy (Torino, Pavia and Verona) and the mutagenic properties of the PM2.5 organic extracts were assessed with the Ames test. The bacterial used in this study were three Salmonella typhimurium strains: TA98, nitroreductase-less mutant TA98NR and YG1021 carrying a nitroreductase-producing plasmid. The annual PM2.5 mean level measured in Torino was 46.5 (+/-31.6) microg/m(3), in Pavia 34.8 (+/-25.1) microg/m(3), and in Verona 37.3 (+/-27.8) microg/m(3), while the mutagenicity expressed as TA98 net reverants/m(3) was 28.0 (+/-22.1), 28.3 (+/-24.9), and 34.2 (+/-30.9) respectively. Monthly pool bioassays, conducted with the three different strains, showed a greater mutagenic response of the YG1021 in each city. The relationship among the mutagenic answers for YG1021:TA98:TA98NR was about 6:3:1 (p<0.001). Over nitroreductase activity enhanced the response of 2.2, 2.0 and 1.7 times for Torino, Pavia, and Verona (ANOVA Torino p<0.05) respectively. Without nitroreductase activity the genotoxicity was limited. These biological findings are able to describe a relevant role played by the nitro compounds in the mutagenic properties of the urban PM2.5 in the Padana plain; moreover the bacterial nitroreductase plays a predominant role in DNA interaction primarily for Torino PM2.5 extracts.

PM2.5 is one of the most important aspects of environmental health. This air pollutant is breathable and it is implicated in several chronic adverse health effects such as the decrease of respiratory functionality and cancer. Several in vitro bioassays are able to predict the mutagenic/carcinogenic activity of the environmental pollutants and mixtures of them. In this study PM2.5 air pollution was daily monitored in three cities located in the Northern part of Italy and the mutagenic properties of the PM2.5 organic extracts were also assessed. Samplings lasted 14 months and cover the period of the Winter Olympic Games of "Torino 2006". In this work, the levels of PM2.5, its mutagenic properties (detected with Salmonella typhimurium assay), the role of the Olympic Games as environmental factor and some meteorological data are discussed. The mean concentration of PM2.5 measured in Torino was 45.4 (+/-30.6) microg/m3, in Pavia 37.6 (+/-25.6) microg/m3, in Verona 43.1 (+/-28.5) microg/m3. Findings of the monthly pool bioassay were in Torino 107 (+/-104) net revertans/m3, in Pavia 108 (+/-89) net revertans/m3, in Verona 128 (+/-109) net revertans/m3. The Olympic Games period data show that PM2.5 pollution and its load of mutagenic potential are different and partially independent phenomena. The Olympic Games had not a great impact on the PM2.5 pollution. The exclusive PM2.5 gravimetric analysis shows a potential human risk if compared with the latest international guide values but it does not describe exhaustively the human health risk associated to the presence of this particular air pollutant. Moreover, the chemical and biological activity qualification of the PM organic extracts as a whole, can instead improve the knowledge.

A preliminary study was performed to evaluate the role of formaldehyde (F) deriving from aspartame intake in the production of the adduct F-human serum albumin (F-HSA) by mean of a sera-epidemiological investigation. A blood-donors population (68 subjects) was analysed for the presence of anti-F-HSA IgG by an indirect competitive immunoenzymatic assay (displacement assay). Only the 41% of the subjects were aspartame consumer and with a low daily intake (0.96mg/(kgday)). A 50% sera-prevalence of IgG anti-F-HSA was observed in the population, but no association between this biomarker and aspartame intake was pointed out. A significant association was found between the IgG anti-F-HSA presence and exogenous F exposure sources (cigarette active smoke and occupational exposure). Considering the low number of the investigated subjects and the low doses of aspartame consumption, the results of this preliminary study seems to suggest that aspartame low intake does not influence the formation of F adducts., (Copyright © 2007 Elsevier B.V. All rights reserved.)

A simplified method to detect faecal sterols, as an alternative assessment of environmental faecal pollution is proposed. The aim of this study is the development of a method to determine sterols in water samples avoiding sample filtration through glass fibre filter. The method is based on a liquid-liquid extraction and a final GC-FID determination. The quantified sterols are coprostanol and 24-ethylcoprostanol, while 5alpha-cholestane is used as internal standard. The recovery of coprostanol and 24-ethylcoprostanol in wastewater ranges from 90 to 100% and the detection limit is 1-2 microgl(-1). Moreover the method proved to be useful for the sterols determination in surface water too.

Benzene (B) is a typical micro-pollutant present in air, especially urban air. In this study a possible correlation between personal benzene exposure and S-phenylmercapturic acid (S-PMA) as a biomarker of internal dose was evaluated in a cohort of traffic policemen. The results confirm that S-PMA is significantly correlated to benzene measured in personal air. B and S-PMA were analyzed considering seasonality, work quarters, time spent indoors, outdoors, and directing traffic, but no significant differences were recorded.

Large segments of populations, including children, are exposed to environmental tobacco smoke (ETS), a risk factor for lung cancer and heart, circulatory and respiratory diseases. Recently, ETS was classified as a class A carcinogen by USEPA, as carcinogenic to humans by IARC (group 1) and by the National Toxicology Program of the US National Institutes of Health. Cotinine, a product of the metabolism of nicotine, is measurable in urine and, correlates strictly and directly to ETS exposure, therefore representing a well-known internal dose marker. Another marker of active tobacco smoking is the N-(2-hydroxyethyl) valine (HOEtVal) which results from the reaction between ethylene oxide (EtO) and the N-terminal valine of hemoglobin. The aim of this study was the evaluations of ETS markers, namely urinary cotinine and HOEtVal measured in blood in 100 children with ages ranging between 3 and 13 years. Experimental findings show that cotinine, as a specific internal dose marker, and HOEtVal, as a nonspecific biological effective dose marker, both depend on the passive exposure to ETS as well as on the active habit of smoking.