Impact of the environment on the health: From theory to practice.

The Erice 56 Charter titled "Impact of the environment on the health: from theory to practice" was unanimously approved at the end of the 56th course of the "International School of Epidemiology and Preventive Medicine G. D'Alessandro" held from 3rd to November 7, 2019 in Erice - Sicily (Italy) and promoted by the Study Group of "Environment and Health" of the Italian Society of Hygiene, Preventive Medicine and Public Health. The course, that included lectures, open discussions and guided working groups, was aimed to provide a general training on epidemiological and toxicological aspects of the environmental health impact, to be used by public health professionals for risk assessment, without forgetting the risk communications. At the end of the course 12 key points were agreed among teachers and students: they underlined the need of specific training and research, in the perspective of "One Health" and "Global Health", also facing emerging scientific and methodological issues and focusing on communication towards stakeholders. This Discussion highlight the need to improve knowledge of Health and Environment topic in all sectors of health and environmental prevention and management.

Water quality and human health: A simple monitoring model of toxic cyanobacteria growth in highly variable Mediterranean hot dry environments.

Due to population growth, urbanization and economic development, demand for freshwater in urban areas is increasing throughout Europe. At the same time, climate change, eutrophication and pollution are affecting the availability of water supplies. Sicily, a big island in southern Italy, suffers from an increasing drought and consequently water shortage. In the last decades, in Sicilian freshwater reservoirs several Microcystis aeruginosa and more recently Planktothrix rubescens blooms were reported. The aims of the study were: (1) identify and quantify the occurring species of cyanobacteria (CB), (2) identify which parameters, among those investigated in the waters, could favor their growth, (3) set up a model to identify reservoirs that need continuous monitoring due to the presences, current or prospected, of cyanobacterial blooms and of microcystins, relevant for environmental and, consequentially, for human health. Fifteen artificial reservoirs among the large set of Sicilian artificial water bodies were selected and examined for physicochemical and microbiological characterization. Additional parameters were assessed, including the presence, identification and count of the cyanobacterial occurring species, the measurement of microcystins (MCs) levels and the search for the genes responsible for the toxins production. Principal Component Analysis (PCA) was used to relate environmental condition to cyanobacterial growth. Water quality was poor for very few parameters, suggesting common anthropic pressures, and PCA highlighted clusters of reservoirs vulnerable to hydrological conditions, related to semi-arid Mediterranean climate and to the use of the reservoir. In summer, bloom was detected in only one reservoir and different species was highlighted among the Cyanobacteria community. The only toxins detected were microcystins, although always well below the WHO reference value for drinking waters (1.0 µg/L). However, molecular analysis could not show the presence of potential cyanotoxins producers since a few numbers of cells among total could be sufficient to produce these low MCs levels but not enough high to be proved by the traditional molecular method applied. A simple environmental risk-based model, which accounts for the high variability of both cyanobacteria growth and cyanotoxins producing, is proposed as a cost-effective tool to evaluate the need for monitoring activities in reservoirs aimed to guarantee supplying waters safety.

Patent method for the extraction and determination of micro- and nano- plastics in organic and inorganic matrix samples: An application on vegetals.

The uncontrolled introduction into the environment of plastic polymers have caused the dispersion of plastic debries, known as microplastics (MPs), that represent an important topic for environmental and human health threats. So far, the absence of effective and efficient extraction methods of MPs (especially for plastic particles with diameters inferior than 10 µm) from complex matrices (water, food, etc.) did not allow to perform the risk estimation and, the consequent assessment of the health impact associated with the exposure to these emergent contaminants. In this paper, a new patented method for the extraction and determination of micro- and nano-plastics in organic and inorganic matrix samples is reported. The method applied in the study has been nationally and internationally protected. The code of the submitted request of international patent's extension in several country of world is PCT/IB2019/051,838 of 7 March 2019, coupled with the accepted Italian patent n. 102,018,000,003,337 of March 7 of 2018 entitled "Method for extraction and determination of microplastics in samples with organic and inorganic matrices". The method applied to our study is based on sedimentation of the particles with density higher than 1 g/cm3. The method can be applied to organic and inorganic samples as water, food, soil, waste, air, biological sample (blood, urine, tissues, etc.). After acid digestion of sample matrix, MPs are recovered by sedimentation in dichloromethane and then they are dispersed in metallic stub. Analysis is performed by SEM-EDX.•New method for the extraction and determination of total microplastics <10 µm.•The method is based on the sedimentation of particles with density higher than 1 g/cm3.•The method can be applied to organic (food, soil, biological sample, etc..) and inorganic samples.•Total micro- and nano-plastics quantification is performed by SEM-EDX.

Reply for comment on "Exposure to microplastics (<10 µm) associated to plastic bottles mineral water consumption: The first quantitative study by Zuccarello et al. [Water Research 157 (2019) 365-371]".

Microplastics (MPs) are a threat to both the environment and human health. In the absence of an official method that would allow the analysis of all MPs with diameters less than 10 µm, it was not possible to estimate the exact exposure to these contaminants, yet. In the last years, few studies are available in literature with the first data concerning the concentrations of MPs on bottle mineral waters. Zuccarello et al. (2019) carried out an exposure assessment study where a high value of EDI (Estimated Daily Intake) has been reported. This rebuttal letter aims to reply to the comments of Oßmann et al. (2018) on the article of Zuccarello et al. (2019) about the new analytical method and both the use and the interpretation of Principal Component Analysis.

Exposure to microplastics (<10 µm) associated to plastic bottles mineral water consumption: The first quantitative study.

The uncontrolled introduction into the environment of plastic polymers have caused the dispersion of plastic fragments, known as Microplastics (MPs), that represent an important topic for public health. This study was the first to investigate the cause of the release of MPs in mineral waters and to estimate the concentration of MPs smaller than 10 µm both in number of particles and in mass unit. This study was carried out using a patent method regarding the extraction and analysis of MPs in more kind of matrix. Therefore, aims of this study were a) to assess the number of MPs with diameters of between 0.5 and 10 µm in mineral waters contained in plastic bottles, b) to evaluate if the physical-chemical properties of mineral waters and bottle quality could influence the release of MPs and, finally, c) to estimate the human daily exposure to MPs due to mineral water consumption. The Mps were found in every sample. The main concentration of MPs was 656.8 µg/L ± 632.9 or 5.42E+07 p/L ± 1.95E+07. The main diameter of detected MPs was 2.44 µm ± 0.66 (where p/L, where p was the number of MPs). The Estimated Daily Intake (EDI) for adults and children were 1,531,524 p/kg/body-weight/day corresponding to 40.1 µg/kg/body-weight/day and 3,350,208 p/kg/body-weight/day corresponding to 87.8 µg/kg/body-weight/day, respectively. The number of MPs contamination in bottled mineral waters was strongly correlated to the pH of waters and to plastic density of bottle. Otherwise, micrograms of MPs per liter and the MPs diameters were strongly affected by plastic thickness. The most mineral water brand contaminated by MPs was the one whose bottles were made from poor quality plastic. In absence of reference values, it was no possible carried out a risk assessment for MPs exposure. It is fundamental to establish the reference method of analysis to monitoring every source of human intake.

Validation of Armadillo officinalis Dumèril, 1816 (Crustacea, Isopoda, Oniscidea) as a bioindicator: in vivo study of air benzene exposure.

This study tests the potential for using Armadillo officinalis as a bioindicator of exposure to and activation of benzene metabolic pathways using an in vivo model. A. officinalis specimens collected in a natural reserve were divided into a control and three test groups exposed to 2.00, 5.32 or 9.09 µg/m(3) benzene for 24h. Three independent tests were performed to assess model reproducibility. Animals were dissected to obtain three pooled tissue samples per group: hepatopancreas (HEP), other organs and tissues (OOT), and exoskeleton (EXO). Muconic acid (MA), S-phenylmercapturic acid (S-PMA), two human metabolites of benzene, and changes in mtDNA copy number, a human biomarker of benzene exposure, were determined in each sample; benzene was determined only in EXO. MA was measured by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection, S-PMA by triple quadrupole mass spectrometer liquid chromatography with electro spray ionization (LC-MS-ESI-TQD), mtDNA by real-time quantitative PCR and end-point PCR, and benzene by quadrupole mass spectrometer head-space gas chromatography (HSGC-MS). MA and S-PMA levels rose both in HEP and OOT; EXO exhibited increasing benzene concentrations; and mtDNA copy number rose in HEP but not in OOT samples. Overall, our findings demonstrate that A. officinalis is a sensitive bioindicator of air benzene exposure and show for the first time its ability to reproduce human metabolic dynamics.

Bioaccumulation of cadmium and lead and its effects on hepatopancreas morphology in three terrestrial isopod crustacean species.

This study was designed to compare cadmium (Cd) and lead (Pb) bioaccumulation in three species of oniscidean isopods - Armadillidium granulatum Brandt, Armadillidium vulgare (Latreille) and Porcellio laevis Latreille which were exposed for three weeks to a contaminated diet, and to determine the morphological and ultrastructural changes in hepatopancreas. Metal accumulation, determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), was linearly associated with the exposed concentration and was a function of the metal and the species tested. All three species accumulated lower levels of Pb than Cd. A. vulgare accumulated the largest concentration of Pb, especially at the higher doses, whereas P. laevis showed the greatest Cd accumulation, and the highest Cd concentration was lethal for all exposed species. The highest concentrations of Pb and Cd induced significant changes both in the general morphology of tubules and in the ultrastructural organization of epithelial cells in hepatopancreas. Some Pb/Cd induced alterations include: brush border disorganization; reduction of the basal labyrinth formed by the plasma membrane; condensation of some cytoplasm areas and of chromatin; rough endoplasmic reticulum and mitochondrial alterations; increase of secondary lysosomes and of type B granules in S cells. Some of the ultrastructural changes observed overlap with those induced by prolonged starvation, whereas others can be useful biomarkers of heavy metal toxicity. This study has confirmed that in terrestrial isopods, the accumulation of the different metals occurs in a species-specific manner; therefore ecological monitoring and assessment studies should consider each species individually. The research has confirmed that in the terrestrial isopods the accumulation of the different metals occurs in a species-specific way; therefore each species should first be evaluated in view of its employ in biomonitoring programs.

Effects of heavy metal accumulation on some reproductive characters in Armadillidium granulatum Brandt (Crustacea, Isopoda, Oniscidea).

The impact of heavy metal bioaccumulation on reproduction of the isopod Armadillidium granulatum was studied by exposing the animals to food contaminated with various sub-lethal concentrations of cadmium, lead and zinc salts over a period of three weeks. The analysis carried out by atomic absorption spectrometry on whole body and on isolated female and male genital systems highlighted that, although metal's bioaccumulation was always concentration-dependent, it varies considerably depending on the metal (Cd>Zn>Pb) as pointed out by the respective values of the concentration factor. The heavy metals bioaccumulation has influenced in different ways the reproductive characters observed; while no significant difference was found with regard to the length of the incubation period and the number of broods--A. granulatum has an iteroparous reproductive strategy--the onset and the length of the reproductive season were negatively affected by the increase in concentration of the tested metals, in particular of Cd. The rate of gravid females, instead, was negatively affected by the bioaccumulation of Cd and Zn while in the groups treated with the highest concentrations of Pb all females produced at least one brood. The number of juveniles released from the brood pouch at the end of incubation resulted considerably higher and it was always positively correlated to the increase of the concentration of each metal, except for the highest Pb concentration. The explanation of this result, apparently anomalous, could be the object of a future research.