Tetrodotoxin in bivalve mollusks: An integrated study towards the comprehension of the influencing factors of a newly native phenomenon.

Tetrodotoxins (TTXs) are potent neurotoxins named after the Tetraodontidae fish family. The ingestion of TTX-contaminated flesh can cause neurotoxic symptoms and can lead to death. In 2017 symptoms the European Food Safety Authority (EFSA) recognized the threat to food safety resulting from TTX exposure via food consumption and, thus, proposed a safety limit of 44 µg/kg of TTX in marine gastropods and bivalves. To date, however, TTXs have not yet been included in the list of biotoxins to be monitored within the European Union, even though, in a few cases, levels of TTX found were higher than the EFSA limit. The origin of TTX production is debated and the roles of both biotic and abiotic factors on TTX-mediated toxic events remain unclear. In order to meet these knowledge requests the present study was aimed to investigate the role of seawater temperature, pH, water conductivity, and oxygen saturation, along with the marine phytoplankton community and the bacterial community of mussels and oysters on the accumulation of TTX and analogues in the bivalves. Abiotic parameters were measured by means of a multi-parametric probe, phytoplankton community was analyzed by optic microscopy while microbial community was described by amplicon metataxonomic sequencing, TTXs concentration in the collected matrices were measured by HILIC-MS/MS. A possible role of seawater pH and temperature, among the investigated abiotic factors, in regulating the occurrence of TTXs was found. Regarding biotic variables, a possible influence of Vibrio, Shewanella and Flavobacteriaceae in the occurrence of TTXs was found. Concurrently, Prorocentrum cordatum cell numbers were correlated to the incidence of TTX in mussels. The results herein collected suggest that environmental variables play a consistent part in the occurrence of TTX in the edible bivalve habitats, and there are also indications of a potential role played by specific bacteria taxa in association with phytoplankton.

Latent Tuberculosis Infection Cascade of Care among Asylum Seekers in Verona, Italy.

Our study aimed to assess latent tuberculosis infection (LTBI) prevalence, screening uptake, adherence to preventive treatment, and their predictors in a large cohort of asylum seekers. We retrospectively analysed data of migrants screened in 2015-2017 at the Migrant's Service in Verona, Italy. Sequential interferon-gamma-release-assay (IGRA) was performed to confirm only tuberculin-skin-test (TST) results ranking from 5 to 14 mm. Among 2,486 asylum seekers, screening adherence was 89.74% and LTBI prevalence was 28.8% (CI95% 27.0;30.5). Predictors of LTBI diagnosis were: male gender (OR 1.62), age 24 years or older (OR 1.47) and African origin (OR 1.78). Therapy completion rate was 69.6% and resulted associated with African origin (OR 1.75) and being older than 24 years (OR 2.89). Sequential IGRA testing, given its expensiveness, could be used to confirm only intermediate TST results, thus enabling further LTBI cases to be detected and avoiding unnecessary preventive treatments.

Tetrodotoxin in live bivalve mollusks from Europe: Is it to be considered an emerging concern for food safety?

Tetrodotoxins (TTXs) are a group of potent neurotoxins named after the Tetraodontidae fish family (pufferfish). TTXs have been reported in several animal taxa, both terrestrial and marine. The ingestion of TTX-contaminated flesh can cause serious neurotoxic symptomatology and can eventually lead to death. Traditionally, TTXs have been associated with Asian countries, in particular with pufferfish consumption. However, they have also been reported in bivalve mollusks farmed in the Pacific area and, recently, in European seas. In Europe, different countries have reported TTXs, especially those bordering the Mediterranean Sea. As a consequence, in 2017 the European Food Safety Authority (EFSA) released an opinion with reference to TTX present in marine gastropods and bivalves, proposing a safety limit of 44 µg/kg TTXs in shellfish meat, below which no adverse effects should be observed in humans. Nevertheless, this limit has been exceeded on many occasions in European shellfish and, while for bivalves there have been no registered human intoxications, that is not the case for marine gastropods. However, TTXs have not yet been included in the list of marine biotoxins officially monitored in live bivalve mollusks within the European Union (EU). Thus, the aims of this manuscript are to discuss the increasing occurrence of TTXs in live bivalve mollusks from European sea waters, to acknowledge the still ongoing knowledge gaps that should be covered and to stimulate constructive debate on the eventuality of adopting a shared regulatory context, at least in the EU, for monitoring and managing this potential threat to food safety.

Knowledge, attitudes and practices about vaccination in Trentino, Italy in 2019.

Introduction: Vaccination is among the most important areas of progress in the worldwide history of public health. However, a crescent wave of anti-vaccine groups has grown in Western countries, especially in Italy, in the last two decades. Our aim was to evaluate adult's hesitancy and knowledge about vaccines and related diseases in Trentino-Alto Adige -the Italian region with the lowest vaccination coverages. Methods: We administered self-answered structured questionnaires in three malls in the Trentino province in June 2019. We collected demographic data and information on knowledge about vaccines, infectious diseases and attitude in seeking health information. We utilized a descriptive and multivariate analysis to investigate factors associated with vaccine hesitancy. Results: We collected 567 questionnaires, 18% of the people interviewed were hesitant toward vaccination and 16% were against mandatory vaccination. In the multivariate analysis a poor level of information, being younger than 60 years and being against compulsory vaccination were associated with vaccine hesitancy. Regarding information about vaccines, 76.5% of the people relied on physicians, and/or 49% navigated the internet, while social media are used by 16% of the study population. Though 41.5% searched information on official sites, only 14% knew the website VaccinarSì and 4.7% had visited it. Discussion: Compared to a previous study conducted in all of Italy except Trentino Alto Adige, the level of vaccination hesitancy was higher. It is important to utilize health professionals, the internet and especially social media to spread scientific information about vaccination.

Health System Response during the European Refugee Crisis: Policy and Practice Analysis in Four Italian Regions.

The decentralization of the provision of health services at the subnational level produces variations in healthcare offered to asylum seekers (ASs) across the different Italian regions, even if they are entitled to healthcare through the national health service. The present study aims to map the healthcare path and regional policies for ASs upon arrival and identify challenges and best practices. This is a multicentric, qualitative study of migrant health policies and practices at the regional level within four Italian regions. For the analysis, a dedicated tool for the systematic comparison of policies and practices was developed. The collection and analysis of data demonstrated the presence of many items of international recommendations, even if many gaps exist and differences between regions remain. The analysis of practices permitted the identification of three models of care and access. Some aspects identified are as follows: fragmentation and barriers to access; a weakness in or lack of a governance system, with the presence of many actors involved; variability in the response between territories. The inclusion of ASs in healthcare services requires intersectoral actions, involving healthcare sectors and other actors within local social structures, in order to add value to local resources and practices, reinforce networks and contribute to social integration.

Molecular Typing of Vibrio parahaemolyticus Strains Isolated from Mollusks in the North Adriatic Sea.

BACKGROUND: Vibrio parahaemolyticus is an emerging foodborne pathogen in the Mediterranean, usually associated with shellfish consumption. The increase in the number of outbreaks in Europe is primarily associated with the global warming of the ocean that has a great impact on the spread and genetic selection of waterborne pathogens. The primary role of Italy in Europe's mollusk production, together with the fact that cases of infections with V. parahaemolyticus are not always notified to the European community, highlighted the necessity of acquiring new information about the epidemiological involvement of shellfish products. OBJECTIVE: The aim of the study was to provide useful insights into the first steps of the Risk Assessment associated with V. parahaemolyticus through the molecular characterization of isolates from commercialized mollusks. MATERIALS AND METHODS: A total of 102 strains identified as V. parahaemolyticus were investigated as part of a larger sampling (1-year survey) from several shellfish species collected from the Venice lagoon and the North Adriatic sea. All strains were characterized by multilocus sequence typing and tested for the presence of virulence genes (trh and tdh). The study of sampling/environmental factors and epidemiological analyses was performed to describe the behaviors of the different genetic populations. RESULTS: The population structure analysis highlighted three genetic clusters that could be subject to temperature selection during cold (≤15°C) and warm (>16°C) seasons. Moreover, other factors, such as molluscan species (clams/mussels), probably played a role in the distribution of genetic clusters. Although few strains carried the virulence factors (n = 6 trh+), epidemiological links with clinical isolates and a local dissemination of some sequence types were underlined. CONCLUSION: This work provides a useful background on the genotype spread as a first step in the Hazard Identification in light of future climate changes.

Effectiveness of Washing Procedures in Reducing Salmonella enterica and Listeria monocytogenes on a Raw Leafy Green Vegetable (Eruca vesicaria).

Vegetables are an important source of nutrients, but they can host a large microbial population, particularly bacteria. Foodborne pathogens can contaminate raw vegetables at any stage of their production process with a potential for human infection. Appropriate washing can mitigate the risk of foodborne illness consequent to vegetable consumption by reducing pathogen levels, but few data are available to assess the efficacy of different practices. In the present work, six different washing methods, in the presence or absence of sanitisers (peracetic acid and percitric acid, sodium bicarbonate, sodium hypochlorite) and vinegar, were tested for their effectiveness in reducing Salmonella and Listeria counts after artificial contamination of raw rocket (Eruca vesicaria). Results showed that washing with sodium hypochlorite (200 mg/L) was the only method able to produce a significant 2 Log reduction of Salmonella counts, but only in the case of high initial contamination (7 Log CFU/g), suggesting potential harmful effects for consumers could occur. In the case of Listeria monocytogenes, all the examined washing methods were effective, with 200 mg/L sodium hypochlorite solution and a solution of peracetic and percitric acids displaying the best performances (2 and 1.5 Log reductions, respectively). This highlights the importance of targeting consumers on fit for purpose and safe washing practices to circumvent vegetable contamination by foodborne pathogens.

Detection and quantification of Vibrio parahaemolyticus in shellfish from Italian production areas.

Vibrio parahaemolyticus is a marine microorganism, recognized as an important cause of foodborne illness particularly in Asia, South America and United States. Outbreaks are rarely reported in Europe, but they can occur unexpectedly in relation, among other reasons, to the spread of highly virulent strains. It is known that the risk is proportional to exposure levels to pathogenic V. parahaemolyticus (i.e. carrying the tdh and/or the trh genes) but currently there is a lack of occurrence data for pathogenic V. parahaemolyticus in shellfish production areas of the Member States. In this study a total of 147 samples of bivalve molluscs, from harvesting areas of two Italian regions (Sardinia and Veneto) were analyzed for Escherichia coli and salmonella, according to Reg 2073/2005, and for detection and enumeration of total and toxigenic V. parahaemolyticus strains using a new DNA colony hybridization method. Environmental parameters (water temperature and salinity) were also recorded. Results of E. coli were consistently in agreement with the legislation limits for the harvesting class of origin and Salmonella was detected only in one sample. The average contamination levels for total V. parahaemolyticus were 84 and 73 CFU/g respectively for Sardinia and Veneto, with the highest value reaching 8.7 × 10(3)CFU/g. Nineteen samples (12.9%) resulted positive for the presence of potentially pathogenic V. parahaemolyticus strains, with levels ranging between 10 and 120 CFU/g and most of the positive samples (n=17) showing values equal or below 20 CFU/g. A significant correlation (r=0.41) was found between water temperature and V. parahaemolyticus levels, as well as with isolation frequency. The data provided in this study on contamination levels of total and potentially pathogenic V. parahaemolyticus, seasonal distribution and correlation with water temperature, will help in defining appropriate monitoring programs and post-harvest policies for this hazard, improving the management of the harvesting areas and the safety of bivalve molluscs.

Vibrio trends in the ecology of the Venice lagoon.

Vibrio is a very diverse genus that is responsible for different human and animal diseases. The accurate identification of Vibrio at the species level is important to assess the risks related to public health and diseases caused by aquatic organisms. The ecology of Vibrio spp., together with their genetic background, represents an important key for species discrimination and evolution. Thus, analyses of population structure and ecology association are necessary for reliable characterization of bacteria and to investigate whether bacterial species are going through adaptation processes. In this study, a population of Vibrionaceae was isolated from shellfish of the Venice lagoon and analyzed in depth to study its structure and distribution in the environment. A multilocus sequence analysis (MLSA) was developed on the basis of four housekeeping genes. Both molecular and biochemical approaches were used for species characterization, and the results were compared to assess the consistency of the two methods. In addition, strain ecology and the association between genetic information and environment were investigated through statistical models. The phylogenetic and population analyses achieved good species clustering, while biochemical identification was demonstrated to be imprecise. In addition, this study provided a fine-scale overview of the distribution of Vibrio spp. in the Venice lagoon, and the results highlighted a preferential association of the species toward specific ecological variables. These findings support the use of MLSA for taxonomic studies and demonstrate the need to consider environmental information to obtain broader and more accurate bacterial characterization.