[Some considerations on revision of legislation Hygiene-sanitary for the management of swimming pools for swimming-pool use and the National Consultation of the Ministry of Health]. / Alcune considerazioni sulla revisione della normativa igienico-sanitaria per la gestione di piscine ad uso natatorio e la Consultazione Nazionale del Ministero della Salute.

After over a decade the Italian regulation on hygiene and surveillance in swimming pools is under revision based on WHO guidelines and current laws from other European countries. The Ministry of Health is proposing an update of the Annex 1 that contains physical, chemical and microbiological indicators for monitoring swimming pool hygiene. The authors review current regulations and the new proposal for updating surveillance. A comparison of Annex 1 as approved in 2003 and the new version as proposed in 2016 allowed to enlighten innovative improvements. The suggested updates underwent a public consultation through an online questionnaire involving citizens and stakeholders, following the required procedure for accessing the final approval of a new regulation. The updates include new requirements and changes involving several controls and microbiological parameters. Safety of recreational waters is presently under revision in different European countries. This updating process is a valuable opportunity to implement critical control points and improve monitoring. The final aim is an optimization of surveillance in terms of efficacy and costs, with an advantage for both the National Health Service as the swimming pool management.

Management of a toxic cyanobacterium bloom (Planktothrix rubescens) affecting an Italian drinking water basin: a case study.

An extraordinary bloom of Planktothrix rubescens, which can produce microcystins (MCs), was observed in early 2009 in the Occhito basin, used even as a source of drinking water in Southern Italy. Several activities, coordinated by a task force, were implemented to assess and manage the risk associated to drinking water contaminated by cyanobacteria. Main actions were: evaluation of analytical protocols for screening and confirmatory purpose, monitoring the drinking water supply chain, training of operators, a dedicated web site for risk communication. ELISA assay was considered suitable for health authorities as screening method for MCs and to optimize frequency of sampling according to alert levels, and as internal control for the water supplier. A liquid chromatography-tandem mass spectrometric method able to quantify 9 MCs was optimized with the aim of supporting health authorities in a comprehensive risk evaluation based on the relative toxicity of different congeners. Short, medium, and long-term corrective actions were implemented to mitigate the health risk. Preoxidation with chlorine dioxide followed by flocculation and settling have been shown to be effective in removing MCs in the water treatment plant. Over two years, despite the high levels of cyanobacteria (up to 160 × 10(6) cells/L) and MCs (28.4 µg/L) initially reached in surface waters, the drinking water distribution was never limited.

An ultra performance liquid chromatography coupled with high resolution mass spectrometry method for the screening of cianotoxins content in drinking water samples.

The incidence of cyanobacterial harmful algal blooms (CHABs) and their potentially toxic secondary metabolites is increasing in Italy and worldwide and several studies demonstrated that the climate change may be playing a role. The method described in this work allows to detect simultaneously 21 cyanotoxins of different classes (including 12 Microcystins, 5 Microginins, 2 Cyanopeptolins, and 2 Anabaenopeptins) in water samples for human consumption by an Ultra Performance Liquid Chromatography coupled with a Q-TOF mass spectrometer. Water samples were freezed, filtered, spiked with Nodularin used as internal standard (I.S.) and then extracted with a SPE Carboghaph 4 cartridge. The extracted sample were analysed injecting 10 µL into the UPLC-HRMS/MS system: the chromatographic separation was obtained using acetonitrile and water as mobile phases, both containing 10 mM formic acid, and a UPLC C18 column, acquiring the experiments in positive ionization and Sensitivity Mode. The described method allows to separate and detect all the selected analytes in short time of analysis (16 minutes) with a good resolution for all the analytes.•simultaneous determination of 21 cyanotoxins belonging to different classes in water sample•low injection volume•short time of analysis.

Multi-residue Ultra Performance Liquid Chromatography-High resolution mass spectrometric method for the analysis of 21 cyanotoxins in surface water for human consumption.

The presence of cyanobacteria and their toxins in water used as drinking water or for recreational purposes may represent a risk for human health. This work describes the development of an advanced analytical method for simultaneous determination of 21 cyanotoxins (including Microcystins, Cyanopeptolins, Anabaenopeptins and Microginins) in drinking water based on Ultra Performance Liquid Chromatography coupled with a Q-TOF mass spectrometer. Water samples, spiked with Nodularin as internal standard at 1 µg/L, were extracted using Carbograph 4 SPE cartridge and 10 µL of the extracted sample were injected into the UPLC-HRMS/MS system. Analytes separation was obtained using a UPLC C18 column, acetonitrile and water as mobile phases, both containing 10 mM formic acid, and operating in positive ionization mode and sensitivity mode. The method has been proven to be robust, precise and accurate with recovery percentages above 85% and with relative standard deviations ≤16% and LODs between 0.002 and 0.047 µg/L, fitting for the intended purposes at the concentrations of interest. This method was applied during a monitoring activity in an Italian volcanic lake in Viterbo (Lazio Region, Italy), due to a severe algal proliferation in January 2018-March 2019 period and for the assessment of cyanobacteria proliferation risk and of cyanotoxin production in drinking water chain.

Quantitative Microbial Risk Assessment as support for bathing waters profiling.

Profiling bathing waters supported by Quantitative Microbial Risk Assessment (QMRA) is key to the WHO's recommendations for the 2020/2021 revision of the European Bathing Water Directive. We developed an area-specific QMRA model on four pathogens, using fecal indicator concentrations (E. coli, enterococci) for calculating pathogen loads. The predominance of illness was found to be attributable to Human Adenovirus, followed by Salmonella, Vibrio, and Norovirus. Overall, the cumulative illness risk showed a median of around 1 case/10000 exposures. The risk estimates were strongly influenced by the indicators that were used, suggesting the need for a more detailed investigation of the different sources of fecal contamination. Area-specific threshold values for fecal indicators were estimated on a risk-basis by modelling the cumulative risk against E. coli and enterococci concentrations. To improve bathing waters assessment, we suggest considering source apportionment, locally estimating of pathogen/indicator ratios, and calculating site-specific indicators thresholds based on risk assessment.

Determination of low-level acrylamide in drinking water by liquid chromatography/tandem mass spectrometry.

A simple and sensitive liquid chromatographic-tandem mass spectrometric (LC/MS/MS) method has been developed and validated to confirm and quantify acrylamide monomer (AA) in drinking water using [13C3] acrylamide as internal standard (IS). After a preconcentration by solid-phase extraction with spherical activated carbon, analytes were chromatographed on IonPac ICE-AS1 column (9 x 250 mm) under isocratic conditions using acetonitrile-water-0.1 M formic acid (43 + 52 + 5, v/v/v) as the mobile phase. Analysis was achieved using a triple-quadrupole mass analyzer equipped with a turbo ion spray interface. For confirmation and quantification of the analytes, MS data acquisition was performed in the multireaction monitoring mode, selecting 2 precursor ion to product ion transitions for both AA and IS. The method was validated for linearity, sensitivity, accuracy, precision, extraction efficiency, and matrix effect. Linearity in tap water was observed over the concentration range 0.1-2.0 microg/L. Limits of detection and quantification were 0.02 and 0.1 microg/L, respectively. Interday and intraday assays were performed across 3 validation levels (0.1, 0.5, and 1.5 microg/L). Accuracy (as mean recovery) ranged from 89.3 to 96.2% with relative standard deviation <7.98%. Performance characteristics of this LC/MS/MS method make it suitable for regulatory confirmatory analysis of AA in drinking water in compliance with European Union and U.S. Environmental Protection Agency standards.

Deriving environmental quality standards for perfluorooctanoic acid (PFOA) and related short chain perfluorinated alkyl acids.

The evidence that in Northern Italy significant sources of perfluoroalkylacids (PFAA) are present induced the Italian government to establish a Working Group on Environmental Quality Standard (EQS) for PFAA in order to include some of them in the list of national specific pollutants for surface water monitoring according to the Water Framework Directive (2000/60/EC). The list of substances included perfluorooctanoate (PFOA) and related short chain PFAA such as perfluorobutanoate (PFBA), perfluoropentanoate (PFPeA), perfluorohexanoate (PFHxA) and perfluorobutanesulfonate (PFBS), which is a substitute of perfluorooctanesulfonate. For each of them a dossier collects available data on regulation, physico-chemical properties, emission and sources, occurrence, acute and chronic toxicity on aquatic species and mammals, including humans. Quality standards (QS) were derived for the different protection objectives (pelagic and benthic communities, predators by secondary poisoning, human health via consumption of fishery products and water) according to the European guideline. The lowest QS is finally chosen as the relevant EQS. For PFOA a QS for biota was derived for protection from secondary poisoning and the corresponding QS for water was back-calculated, obtaining a freshwater EQS of 0.1µgL-1. For PFBA, PFPeA, PFHxA and PFBS threshold limits proposed for drinking waters were adopted as EQS.

Confirmatory analysis of 17beta-boldenone, 17alpha-boldenone and androsta-1,4-diene-3,17-dione in bovine urine by liquid chromatography-tandem mass spectrometry.

A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for confirmatory analysis of 17beta-boldenone (17beta-BOL), 17alpha-boldenone (17alpha-BOL) and androsta-1,4-diene-3,17-dione (ADD) in bovine urine was developed. [2H(2)]17beta-Testosterone (17beta-T-d(2)) was used as the internal standard. Sample preparation involved enzymatic hydrolysis and purification on a C(18) solid-phase extraction column. Chromatographic separation of the analytes was obtained using an RP-C(18) HPLC column. LC-MS-MS detection was carried out with an atmospheric pressure chemical ionisation (APCI) source equipped with a heated nebulizer (HN) interface operating in the positive ion mode. For unambiguous hormone confirmation, three analyte precursor-product ion combinations were monitored during multiple-reaction monitoring (MRM) LC-MS-MS analysis. Overall recovery (%), repeatability (relative standard deviations, RSD, %) and within-laboratory reproducibility (RSD, %) ranged from 92.2 to 97.7%, from 6.50 to 2.94% and from 13.50 to 5.04%, respectively, for all analytes. The limit of quantification in bovine urine was 0.20 ng ml(-1) for 17beta-BOL and ADD and 0.50 ng ml(-1) for 17alpha-BOL. The validated method was successfully applied for determination of 17beta-BOL, 17alpha-BOL and ADD in a large number of bovine urine samples collected within the national Official Residue Control Program.

Quantitative determination of sufentanil in human plasma by liquid chromatography-tandem mass spectrometry.

A sensitive and specific method for the quantification of sufentanil in human plasma by liquid chromatography coupled with tandem mass spectrometry has been developed. Fentanyl was used as the internal standard. Rapid sample preparation involved purification on a C(18) solid-phase extraction column. Chromatographic separation of the analytes was obtained using an RP-C(18) mu-HPLC column. LC-MS-MS detection was performed by atmospheric pressure ionisation (API) source equipped with an ionspray (IS) interface operating in the positive ion mode. For unambiguous substance confirmation, three analyte precursor-product ion combinations were monitored during multiple reaction monitoring (MRM) LC-MS-MS analysis. The method's performance characteristics were evaluated in blank and spiked control plasma samples. Overall accuracy (relative error, R.E., %), repeatability (relative standard deviations, R.S.D., %) and within-laboratory reproducibility (R.S.D., %) ranged from -9.28 to -2.71%, from 6.42 to 2.82% and from 13.52 to 6.06%, respectively, for sufentanil. The limit of quantification for sufentanil in human plasma samples was 0.3 ng ml(-1). Due to its high sensitivity and specificity, the method was successfully employed for sufentanil determination in maternal plasma samples collected immediately before epidural administration of a single sufentanil dose to women in labour, 20 min after drug administration, and at birth in arterial and venous umbilical cord plasma samples from the newborn babies. Research is in progress to adopt the method for performance of complete pharmacokinetic studies of sufentanil in human plasma.

A case study of sanitary survey on community drinking water supplies after a severe (post-Tsunami) flooding event.

This report presents a case study of a comprehensive sanitary survey on ca. 160 community drinking water supplies after a severe (post-Tsunami) flooding event in Sri Lanka. Sanitary inspection and microbiological and chemical water quality analyses were performed according to specifically-designed procedures established on the World Health Organization (WHO) guidelines. Significant hazards and critical points were identified in almost all the investigated water supplies. The overall results showed a significant level of microbiological and chemical risk associated with drinking water consumption within the investigated areas. The criteria and methods practised in this study are proposed as a model to assure an effective and reliable monitoring in post-emergencies involving possible deterioration of water quality and to identify health priorities related to water consumption.