A systematic scoping review of antibiotic-resistance in drinking tap water.

Environmental matrices have been considered of paramount importance in the spread of antibiotic-resistance; however, the role of drinking waters is still underexplored. Therefore, a scoping review was performed using a systematic approach based on PRISMA guidelines, with the aim of identifying and characterizing antibiotic-resistance in tap water, specifically, water treated at a potabilization plant and provided for drinking use through a water distribution system. The review included 45 studies, the majority of which were conducted in upper-middle-income economies (42.2%), mainly from the Western Pacific region (26.7%), followed by Europe (24.4%). Most of the papers focused on detecting antibiotic-resistant bacteria (ARB), either alone (37.8%) or in combination with antibiotic-resistant genes (ARGs) (26.7%). Multidrug-resistance profile was often identified in heterotrophic bacteria, including various species of nontuberculous mycobacteria, Pseudomonas spp., and Aeromonas spp., which were especially resistant to penicillins, cephalosporins (including 3rd-generation), and also to macrolides (erythromycin) and tetracyclines. Resistance to a wide range of antibiotics was also prevalent in fecal bacteria, e.g., the Enterobacteriaceae family, with common resistance to (fluoro)quinolones and sulfonamide groups. ARGs were investigated either in bacterial strains isolated from tap waters or directly in water samples, and the most frequently detected ARGs belonged to ß-lactam, sulfonamide, and tetracycline types. Additionally, mobile genetic elements were found (i.e., int1 and tnpA). Sulfonamides and macrolides were the most frequently detected antibiotics across countries, although their concentrations were generally low (<10 ng/L) in Europe and the United States. From a health perspective, tap water hosted ARB of health concern based on the 2024 WHO bacterial priority pathogens list, mainly Enterobacteriaceae resistant to 3rd-generation cephalosporin and/or carbapenem. Despite the fact that tap water is treated to meet chemical and microbiological quality standards, current evidence suggests that it can harbor antibiotic-resistance determinants, thus supporting its potential role in environmental pathways contributing to antibiotic resistance.

Genotoxic effects of particulate matter on larvae of a common and widespread butterfly along an urbanization gradient.

Biodiversity is currently declining worldwide. Several threats have been identified such as habitat loss and climate change. It is unknown if and how air pollution can work in addition or in synergy to these threats, contributing to the decline of current species and/or local extinction. Few studies have investigated the effects of particulate matter (PM), the main component of air pollution, on insects, and no studies have investigated its genotoxic effects through Micronucleus assay. Butterflies play an important role in the environment, as herbivores during larval stages, and as pollinators as adults. The aim of this study was to evaluate the genotoxic effects of PM10 from different sites along a gradient of population urbanization, on a common cabbage butterfly species (Pieris brassicae). PM10 was collected from April to September in an urban (Turin, Italy), a suburban (Druento, Italy) and a mountain site (Ceresole Reale, Italy) with different urbanization levels. P. brassicae larvae (n = 218) were reared in the laboratory under controlled conditions (26 °C, L:D 15:9) on cabbage plants (average 9.2 days), and they were exposed to PM10 organic extracts (20 and 40 m3/mL) or dimethyl sulfoxide (controls) through vaporization. After exposure, larvae were dissected and cells were used for the Micronucleus (MN) assay. Results showed that all PM extracts induced significant DNA damage in exposed larvae compared to controls, and that increasing the PM dose (from 20 to 40 m3/mL) increased genotoxic effects. However, we did not detect any significant differences between sites with different urbanization levels. In conclusion, PM at different concentrations induced genotoxic effects on larvae of a common butterfly species. More alarmingly, PM could work in addition to and/or in synergy with other compounds (e.g. pesticides) and, especially on species already threatened by other factors (e.g. fragmentation), thus affecting the vitality of populations, leading to local extinctions.

Cytotoxicity and genotoxicity of light emitted by incandescent, halogen, and LED bulbs on ARPE-19 and BEAS-2B cell lines.

LED technology has the extraordinary ability to reduce energy consumption, constituting an economic and ecological advantage, so it is planned to replace incandescent, halogen and other inefficient bulbs for public and domestic lighting with LEDs. LEDs present specific spectral and energetic characteristics compared with those of other domestic light sources, so the potential risks for human health of these bulbs need to be explored. The aim of this study was to assess cytotoxicity and genotoxicity of light emitted by different commercial light bulbs: incandescent, halogen, and two LED bulbs with different correlated color temperatures. The evaluation was done on ARPE-19 as a specific cell model for eye toxicity and on BEAS-2B as a good cell model for toxicology tests. Light induced mainly cytotoxic effects on ARPE-19 and DNA damage on BEAS-2B, so different cell lines showed different biological responses. Moreover, our findings indicate that among the four bulbs, cold LED caused the highest cytotoxic effect on ARPE-19 and the highest genotoxic and oxidative effect on BEAS-2B. Cold LED is probably able to cause more cellular damage because it contains more high-energy radiations (blue). These results suggest that LED technology could be a safe alternative to older technologies, but the use of warm LED should be preferred to cold LED, which can potentially cause adverse effects on retinal cells.

Lifestyles and socio-cultural factors among children aged 6-8 years from five Italian towns: the MAPEC_LIFE study cohort.

BACKGROUND: Lifestyles profoundly determine the quality of an individual's health and life since his childhood. Many diseases in adulthood are avoidable if health-risk behaviors are identified and improved at an early stage of life. The aim of the present research was to characterize a cohort of children aged 6-8 years selected in order to perform an epidemiological molecular study (the MAPEC_LIFE study), investigate lifestyles of the children that could have effect on their health status, and assess possible association between lifestyles and socio-cultural factors. METHODS: A questionnaire composed of 148 questions was administered in two different seasons to parents of children attending 18 primary schools in five Italian cities (Torino, Brescia, Pisa, Perugia and Lecce) to obtain information regarding the criteria for exclusion from the study, demographic, anthropometric and health information on the children, as well as some aspects on their lifestyles and parental characteristics. The results were analyzed in order to assess the frequency of specific conditions among the different seasons and cities and the association between lifestyles and socio-economic factors. RESULTS: The final cohort was composed of 1,164 children (50.9 boys, 95.4% born in Italy). Frequency of some factors appeared different in terms of the survey season (physical activity in the open air, the ways of cooking certain foods) and among the various cities (parents' level of education and rate of employment, sport, traffic near the home, type of heating, exposure to passive smoking, ways of cooking certain foods). Exposure to passive smoking and cooking fumes, obesity, residence in areas with heavy traffic, frequency of outdoor play and consumption of barbecued and fried foods were higher among children living in families with low educational and/or occupational level while children doing sports and consuming toasted bread were more frequent in families with high socio-economic level. CONCLUSIONS: The socio-economic level seems to affect the lifestyles of children enrolled in the study including those that could cause health effects. Many factors are linked to the geographical area and may depend on environmental, cultural and social aspects of the city of residence.

Green job bio-aerosol exposure during anaerobic digestion for biomass energetic valorisation.

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.

Antibiotic resistance and pathogen spreading in a wastewater treatment plant designed for wastewater reuse.

Climate change significantly contributes to water scarcity in various regions worldwide. While wastewater reuse is a crucial strategy for mitigating water scarcity, it also carries potential risks for human health due to the presence of pathogenic and antibiotic resistant bacteria (ARB). Antibiotic resistance represents a Public Health concern and, according to the global action plan on antimicrobial resistance, wastewater role in selecting and spreading ARB must be monitored. Our aim was to assess the occurrence of ARB, antibiotic resistance genes (ARGs), and potential pathogenic bacteria throughout a wastewater treatment plant (WWTP) designed for water reuse. Furthermore, we aimed to evaluate potential association between ARB and ARGs with antibiotics and heavy metals. The results obtained revealed the presence of ARB, ARGs and pathogenic bacteria at every stage of the WWTP. Notably, the most prevalent ARB and ARG were sulfamethoxazole-resistant bacteria (up to 7.20 log CFU mL-1) and sulII gene (up to 5.91 log gene copies mL-1), respectively. The dominant pathogenic bacteria included Arcobacter, Flavobacterium and Aeromonas. Although the abundance of these elements significantly decreased during treatment (influent vs. effluent, p < 0.05), they were still present in the effluent designated for reuse. Additionally, significant correlations were observed between heavy metal concentrations (copper, nickel and selenium) and antibiotic resistance elements (ampicillin-resistant bacteria, tetracycline-resistant bacteria, ARB total abundance and sulII) (p < 0.05). These results underscore the importance of monitoring the role of WWTP in spreading antibiotic resistance, in line with the One Health approach. Additionally, our findings suggest the need of interventions to reduce human health risks associated with the reuse of wastewater for agricultural purposes.

Can oestrogenic activity in air contribute to the overall body burden of endocrine disruptors?

Endocrine disruptors (EDCs) are emerging contaminants that are harmful to health. Human exposure occurs mainly through ingestion or dermal contact, but inhalation could be an additional exposure route; therefore, this study was conducted to evaluate the oestrogenic activity of airborne particulate matter (PM). Outdoor PM was collected for a year in five Italian sites and extracted with organic solvents (four seasonal extracts/site). The oestrogenic activity was assessed using a gene reporter assay (MELN), and the risk to human health through inhalation was quantified using the results. Moreover, extracts were analysed to assess cytotoxicity (WST-1 and LDH assays) on human bronchial cells (BEAS-2B). The extracts induced a significant cytotoxicity and oestrogenic activity. Oestrogenic activity showed a seasonal trend and was correlated with concentrations of benzo(a)pyrene and toxic equivalency factor. Although a low inhalation cancer risk was found, this study confirmed that oestrogenic activity in air could contribute to overall health risks due to EDC exposure.

Biological effects of particulate matter samples during the COVID-19 pandemic: a comparison with the pre-lockdown period in Northwest Italy.

In 2020, during the COVID-19 pandemic, containment measures were applied inducing potential changes in air pollutant concentrations and thus in air toxicity. This study evaluates the role of restrictions on biological effects of particulate matter (PM) in different Northwest Italy sites: urban background, urban traffic, rural, and incinerator. Daily PM samples collected in 2020 were pooled according to restrictions: January/February (no restrictions), March and April (first lockdown), May/June and July/August/September (low restrictions), October/November/December (second lockdown). The 2019 samples (pre-pandemic period) were pooled as 2020 for comparison. Pools were extracted with organic solvents and extracts were tested to assess cytotoxicity (WST-1 assay) and genotoxicity (comet assay) on BEAS-2B cells, mutagenicity (Ames test) on TA98 and TA100 Salmonella typhimurium strains, and estrogenic activity (gene reporter assay) on MELN cells. Pollutant concentrations were also analyzed (PM10, PM2.5, polycyclic aromatic hydrocarbons). No difference was observed for PM and polycyclic aromatic hydrocarbon concentrations between 2020 and 2019. During lockdown months (2020), PM cytotoxicity/genotoxicity was significantly lower in some sites than during 2019, while considering PM mutagenicity/estrogenic activity some differences were detected but without statistical significance. PM extract effects decreased in some sites during 2020; this may be due to lockdowns that reduced/modified pollutant emissions and may be related also to complex PM origin/formation and to meteorological conditions. In conclusion, the study confirms that PM biological effects cannot be assessed considering only the PM concentration and suggests to include a battery of bioassay for air quality monitoring in order to protect human health from air pollution effects. Supplementary Information: The online version contains supplementary material available at 10.1007/s11869-023-01381-6.

Cabbage butterfly as bioindicator species to investigate the genotoxic effects of PM10.

Atmospheric pollution poses a serious threat to environment and human health, and particulate matter (PM) is one of the major contributors. Biological effects induced by PM are investigated through in vitro assays using cells and by in vivo tests with laboratory model animals. However, also the estimation of adverse effects of pollutants, including airborne ones, on wild animals, such as insects, is an essential component of environmental risk assessment. Among insects, butterflies are sensitive to environmental changes and are important wild pollinators, so they might be suitable as environmental bioindicator species. The aim of this study was to evaluate the suitability of a wild cabbage butterfly species (Pieris brassicae) as a bioindicator organism to assess the genotoxic effects of PM10 collected in different sites. PM10 was collected from April to September in urban, suburban, and rural sites. P. brassicae larvae were reared in laboratory under controlled conditions on cabbage plants and exposed to PM10 organic extracts or dimethyl sulfoxide (controls) through vaporization. After exposure, larvae were dissected, and cells were used for comet assay. All PM extracts induced significant DNA damage in exposed larvae compared to controls and the extract collected in the most polluted site caused the highest genotoxic effect. In conclusion, the study suggested that butterflies, such as P. brassicae, could be applied as sensitive and promising bioindicators to investigate air quality and PM genotoxicity. Indeed, the use of these organisms allows the detection of genotoxic effects induced by PM sampled also in low-polluted areas.

Occurrence of antibiotic-resistant bacteria and resistance genes in the urban water cycle.

This study investigates the antibiotic resistance fate in the urban water cycle, evaluating the dynamics of antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs) in three different full-scale wastewater treatment plants (WWTPs) and two drinking water treatment plants (DWTPs) located in the same geographical area (North-West of Italy). ARB (tetracycline-, ampicillin-, and sulfonamide-resistant bacteria) were quantified by plate counting and the abundances of selected ARGs (i.e., tetA, blaTEM, and sulII) and intI1 gene were measured using quantitative real-time PCR (qPCR). Higher concentrations of ARB and ARGs were observed in the WWTPs with respect to the DWTPs identifying the WWTP as hotspot for the spread of antibiotic resistances. Although a significant reduction of ARB and ARGs was observed in WWTPs and DWTPs after the treatment, none of the detected ARB or ARGs was completely removed in drinking water. The stability of the antibiotic-resistant rates between inlet and outlet associated with the reduction of relative ARG abundances underlined that both the treatments (WWTs and DWTs) did not apply any selective pressure. The overall results highlighted the importance to investigate the antibiotic resistance dynamics in aquatic ecosystems involved in urban water cycle integrating the information obtained by culture-dependent method with the culture-independent one and the need to monitor the presence of ARB and ARGs mainly in drinking water that represents a potential route of transmission to human.

A Biomonitoring Pilot Study in Workers from a Paints Production Plant Exposed to Pigment-Grade Titanium Dioxide (TiO2).

Among particulate matter composing paints, titanium dioxide (TiO2) forms about 20% of the final suspension. Although TiO2 is broadly used in many applications, TiO2 powders represent an established respiratory hazard for workers with long-term exposure. In 35 workers of a paints production plant (15 exposed and 20 not exposed), we assessed pro-inflammatory cytokines (IL-1ß, TNF-α, IL-10, IL-17), surfactant protein D (SP-D) and Krebs von den Lungen-6 glycoprotein (KL-6) in exhaled breath condensate (EBC). In urine samples, we measured 8-isoprostane (Isop) and Malondialdehyde (MDA) as biomarkers of oxidative stress, and Titanium (Ti-U) as a biomarker of exposure. Health status, habits and occupational history were recorded. Airborne respirable dusts and Ti were quantified. Particle number concentration and average diameter (nm) were detected by a NanoTracer™ monitoring device. Ti was measurable in filters collected at the respiratory breathing zone (0.11−0.44 µg/m3 8-h TWA). IL-1ß and IL-10 values were significantly higher in exposed workers, whereas SP-D was significantly lower (p < 0.001). KL-6 was significantly higher in workers than in controls (p < 0.01). MDA levels were significantly increased in exposed workers and were positively correlated with Ti-U. Exposure to TiO2 in paint production is associated with the subtle alterations of lung pathobiology. These findings suggest the need for an integrated approach relying on both personal exposure and biomarker assessment to improve the hazard characterisation in occupational settings.

Development of a PCR protocol for the detection of Escherichia coli O157:H7 and Salmonella spp. in surface water.

Escherichia coli O157:H7 and Salmonella are pathogenic microorganisms that can cause severe gastrointestinal illness in humans. These pathogens may be transmitted in a variety of ways, including food and water. The presence of Salmonella and E. coli O157:H7 in surface waters constitutes a potential threat to human health when used for either drinking or recreation. As with most waterborne pathogens, Salmonella and E. coli O157:H7 are difficult to detect and enumerate with accuracy in surface waters due to methodological limitations. The aim of this study was to develop a protocol for the detection of Salmonella spp., E. coli O157:H7 and E. coli virulence genes (stx (1), stx (2) and eae) in water using a single enrichment step and PCR. In spiked water samples, PCR results showed high sensitivity (<3 CFU/L) for both microorganisms. The protocol developed in this study has been applied in different surface waters in association with microbiological and physical analysis. The frequency of PCR positive samples was 33% for Salmonella and 2% for E. coli O157:H7 producing intimin (eae) and Shiga-like toxin I (stx (1)). Moreover, the finding of amplicons corresponding to eae and stx (1) genes in the absence of E. coli O157:H7 suggested the possible presence of other pathogenic bacteria that carry these genes (e.g. EHEC, Shigella strains). The results obtained showed that the developed protocol could be applied as a routine analysis of surface water for the evaluation of microbiological risks.

Comparison of UV, Peracetic Acid and Sodium Hypochlorite Treatment in the Disinfection of Urban Wastewater.

One source of water contamination is the release of wastewater that has not undergone efficient treatment. The aim of this study was to evaluate the reduction obtained with sodium hypochlorite (NaClO), UV and peracetic acid disinfection treatment of Salmonella spp., pathogenic Campylobacter, STEC and bacterial indicators in three full-scale municipal wastewater plants. A general reduction in Salmonella was observed after disinfection, but these bacteria were detected in one UV-treated sample (culture method) and in 33%, 50% and 17% of samples collected after NaClO, UV and PAA disinfection treatments, respectively (PCR method). A better reduction was also observed under NaClO disinfection for the microbial indicators. Independent of the disinfection treatment, E. coli O157:H7 was not detected in the disinfected samples, whereas some samples treated with UV and PAA showed the presence of the stx1 gene. No reduction in the presence of stx2 genes was verified for any of the disinfection treatments. Campylobacter was not detected in any of the analysed samples. The overall results highlight a better reduction in microbiological parameters with a NaClO disinfection treatment in a full-scale municipal wastewater plant compared with UV and PAA. However, the results indicate that a complete and specific monitoring program is necessary to prevent a possible risk to public health.

In Vitro Effects of Particulate Matter Associated with a Wildland Fire in the North-West of Italy.

Wildland fires, increasing in recent decades in the Mediterranean region due to climate change, can contribute to PM levels and composition. This study aimed to investigate biological effects of PM2.5 (Ø < 2.5 µm) and PM10 (Ø < 10 µm) collected near a fire occurred in the North-West of Italy in 2017 and in three other areas (urban and rural areas). Organic extracts were assessed for mutagenicity using Ames test (TA98 and TA100 strains), cell viability (WST-1 and LDH assays) and genotoxicity (Comet assay) with human bronchial cells (BEAS-2B) and estrogenic activity using a gene reporter assay (MELN cells). In all sites, high levels of PM10 and PM2.5 were measured during the fire suggesting that near and distant sites were influenced by fire pollutants. The PM10 and PM2.5 extracts induced a significant mutagenicity in all sites and the mutagenic effect was increased with respect to historical data. All extracts induced a slight increase of the estrogenic activity but a possible antagonistic activity of PM samples collected near fire was observed. No cytotoxicity or DNA damage was detected. Results confirm that fires could be relevant for human health, since they can worsen the air quality increasing PM concentrations, mutagenic and estrogenic effects.

Editorial Comments to the Special Issue: "Legionella Contamination in Water Environment".

Legionella spp [...].

Cryptosporidium Oocyst Contamination in Drinking Water: A Case Study in Italy.

The aim of this study was to evaluate the occurrence of Cryptosporidium oocysts in a drinking water treatment plant (DWTP) located in a rural area of northern Italy. Influent and effluent samples were collected at the DWTP over three years (2013-2016). In parallel, tap water samples from a public drinking fountain were collected as well. All samples were analyzed for the presence of Cryptosporidium spp. oocysts by a common method based on an immunomagnetic separation (IMS)/immunofluorescence assay (IFA), complemented by 4,6-diamidino-2-phenylindole (DAPI) staining. A reverse transcriptase-PCR (RT-PCR) protocol was added to evaluate oocyst viability. The results highlighted a high variability of oocyst concentrations across all samples (mean 4.3 ± 5.8/100 L) and a high variability in the percentage of DAPI-positive specimens (mean 48.2% ± 40.3%). Conversely, RT-PCR did not reveal the presence of viable C. parvum and C. hominis oocysts. A nested PCR targeting Cryptosporidium 18S ribosomal DNA, carried out in two water samples, confirmed the presence of a Cryptosporidium genotype associated with wild animals in the river and in tap water. The results obtained underline the vulnerability of the investigated surface water to Cryptosporidium spp. contamination. Although the recovered Cryptosporidium genotype is not a human pathogen, its presence demonstrates the existence of a potential pathogen Cryptosporidium spp. contamination risk. Moreover, these results underline the importance of also considering unconventional (not bacterial) biological contaminations (protozoa) in water resources in rural areas, including those of developed countries.

Shape-engineered titanium dioxide nanoparticles (TiO2-NPs): cytotoxicity and genotoxicity in bronchial epithelial cells.

The aim of this study was to evaluate cytotoxicity (WST-1 assay), LDH release (LDH assay) and genotoxicity (Comet assay) of three engineered TiO2-NPs with different shapes (bipyramids, rods, platelets) in comparison with two commercial TiO2-NPs (P25, food grade). After NPs characterization (SEM/T-SEM and DLS), biological effects of NPs were assessed on BEAS-2B cells in presence/absence of light. The cellular uptake of NPs was analyzed using Raman spectroscopy. The cytotoxic effects were mostly slight. After light exposure, the largest cytotoxicity (WST-1 assay) was observed for rods; P25, bipyramids and platelets showed a similar effect; no effect was induced by food grade. No LDH release was detected, confirming the low effect on plasma membrane. Food grade and platelets induced direct genotoxicity while P25, food grade and platelets caused oxidative DNA damage. No genotoxic or oxidative damage was induced by bipyramids and rods. Biological effects were overall lower in darkness than after light exposure. Considering that only food grade, P25 and platelets (more agglomerated) were internalized by cells, the uptake resulted correlated with genotoxicity. In conclusion, cytotoxicity of NPs was low and affected by shape and light exposure, while genotoxicity was influenced by cellular-uptake and aggregation tendency.

Mutagenic and genotoxic effects induced by PM0.5 of different Italian towns in human cells and bacteria: The MAPEC_LIFE study.

Particulate matter (PM) is considered an atmospheric pollutant that mostly affects human health. The finest fractions of PM (PM2.5 or less) play a major role in causing chronic diseases. The aim of this study was to investigate the genotoxic effects of PM0.5 collected in five Italian towns using different bioassays. The role of chemical composition on the genotoxicity induced was also evaluated. The present study was included in the multicentre MAPEC_LIFE project, which aimed to evaluate the associations between air pollution exposure and early biological effects in Italian children. PM10 samples were collected in 2 seasons (winter and spring) using a high-volume multistage cascade impactor. The results showed that PM0.5 represents a very high proportion of PM10 (range 10-63%). PM0.5 organic extracts were chemically analysed (PAHs, nitro-PAHs) and tested by the comet assay (A549 and BEAS-2B cells), MN test (A549 cells) and Ames test on Salmonella strains (TA100, TA98, TA98NR and YG1021). The highest concentrations of PAHs and nitro-PAHs in PM0.5 were observed in the Torino, Brescia and Pisa samples in winter. The Ames test showed low mutagenic activity. The highest net revertants/m3 were observed in the Torino and Brescia samples (winter), and the mutagenic effect was associated with PM0.5 (p < 0.01), PAH and nitro-PAH (p < 0.05) concentrations. The YG1021 strain showed the highest sensitivity to PM0.5 samples. No genotoxic effect of PM0.5 extracts was observed using A549 cells except for some samples in winter (comet assay), while BEAS-2B cells showed light DNA damage in the Torino, Brescia and Pisa samples in winter, highlighting the higher sensitivity of BEAS-2B cells, which was consistent with the Ames test (p < 0.01). The results obtained showed that it is important to further investigate the finest fractions of PM, which represent a relevant percentage of PM10, taking into account the chemical composition and the biological effects induced.

Detection of biogenic amine producer bacteria in a typical Italian goat cheese.

The aim of this study was to research decarboxylating bacterial strains and biogenic amine content in a typical Italian goat cheese (Robiola di Roccaverano). The study was performed on fresh and ripened samples of goat cheese manufactured from industrial and artisanal producers. Sixty-seven bacterial strains isolated showed decarboxylating activity, and Enterococcus faecalis was the most widespread decarboxylating species in all artisanal and industrial products. Pediococcus acidilactici and Enterococcus malodoratus were also identified as biogenic amine producers in Robiola di Roccaverano cheese. All the E. faecalis strains isolated in this study were able to decarboxylate tyrosine. Tyramine was the most abundant biogenic amine in cheese samples, while histamine was the most widespread. High amounts of these two biogenic amines were found in ripened samples (up to 2,067 mg/kg for tyramine and 1,786 mg/kg for histamine), whereas 2-phenylethylamine and tryptamine were present in almost all ripened cheeses at low concentrations. The detection of strains producing biogenic amines and the high concentrations of tyramine and histamine found in ripened Robiola di Roccaverano could represent a potential risk to the consumer.

Microbiological characterisation of Robiola di Roccaverano cheese using PCR-DGGE.

The aim of this study was the microbiological characterisation of a typical Italian cheese "Robiola di Roccaverano" with Protected Designation of Origin (PDO). Fresh and ripened robiola samples were collected from four artisanal and one industrial producer. Artisanal producers used raw goat's milk and natural fermentation, whilst the industrial producer used mixed cow-goat's milk and selected starters. The microbial communities were monitored during different seasons and ripening times with PCR-DGGE and culture-dependent methods. The cluster analysis showed that the DGGE bacterial patterns were related to the different manufacturing and climatic conditions, revealing the occurrence of species associated to Robiola di Roccaverano. The DGGE profiles of yeasts were affected by ripening in summer season. Moreover, the results obtained allowed the identification of microbial species such as Lactococcus lactis subsp. lactis, Geotricum spp. and Kluyveromyces lactis that are related to the production of this typical cheese.