Possible Influence of Natural Events on Heavy Metals Exposure from Shellfish Consumption: A Case Study in the North-East of Italy

The objective of this study was the estimation of the exposure over time to heavy metals (cadmium, mercury and lead) due to shellfish consumption in the Veneto Region, Italy. Shellfish consumption was investigated by a food frequency consumption survey. Altogether, 1949 households, stratified into the five most populated areas of the Veneto Region, were involved in the study. Exposure estimation to heavy metals was carried out taking into account the level of metal measured in samples of Manila clams (Ruditapes philippinarum) and grooved carpetshell (Ruditapes decussatus), collected in the frame of the monitoring activities of mollusk production areas of Veneto Region, between January 2007 and December 2012. A general high contribution of the considered shellfish to the Tolerable Weekly Intake (TWI) was noticed in the case of cadmium, especially in 2011, when a considerable increase in cadmium intake was estimated. This was probably due to a heavy rainfall event that triggered catastrophic flooding with high impact on shellfish capture areas in November 2010. The results strongly emphasise the importance of dealing with food safety in a holistic way, taking into account the potential impact of extraordinary natural events on food chain contamination, in order to identify food hazards at an early stage, before developing into a real risk for consumers

Mitochondrial Features of Mouse Myoblasts Are Finely Tuned by Low Doses of Ozone: The Evidence In Vitro

The mild oxidative stress induced by low doses of gaseous ozone (O3) activates the antioxidant cell response through the nuclear factor erythroid 2-related factor 2 (Nrf2), thus inducing beneficial effects without cell damage. Mitochondria are sensitive to mild oxidative stress and represent a susceptible O3 target. In this in vitro study, we investigated the mitochondrial response to low O3 doses in the immortalized, non-tumoral muscle C2C12 cells; a multimodal approach including fluorescence microscopy, transmission electron microscopy and biochemistry was used. Results demonstrated that mitochondrial features are finely tuned by low O3 doses. The O3 concentration of 10 μg maintained normal levels of mitochondria-associated Nrf2, promoted the mitochondrial increase of size and cristae extension, reduced cellular reactive oxygen species (ROS) and prevented cell death. Conversely, in 20 μg O3-treated cells, where the association of Nrf2 with the mitochondria drastically dropped, mitochondria underwent more significant swelling, and ROS and cell death increased. This study, therefore, adds original evidence for the involvement of Nrf2 in the dose-dependent response to low O3 concentrations not only as an Antioxidant Response Elements (ARE) gene activator but also as a regulatory/protective factor of mitochondrial function

Molecular Evolution and Phylogeography of Co-circulating IHNV and VHSV in Italy

Infectious haematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV) are the most important viral pathogens impacting rainbow trout farming. These viruses are persistent in Italy, where they are responsible for severe disease outbreaks (epizootics) that affect the profitability of the trout industry. Despite the importance of IHNV and VHSV, little is known about their evolution at a local scale, although this is likely to be important for virus eradication and control. To address this issue we performed a detailed molecular evolutionary and epidemiological analysis of IHNV and VHSV in trout farms from northern Italy. Full-length glycoprotein gene sequences of a selection of VHSV (n=108) and IHNV (n=89) strains were obtained. This revealed that Italian VHSV strains belong to sublineages Ia1 and Ia2 of genotype Ia and are distributed into 7 genetic clusters. In contrast, all Italian IHNV isolates fell within genogroup E, for which only a single genetic cluster was identified. More striking was that IHNV has evolved more rapidly than VHSV (mean rates of 11 and 7.3 × 10-4 nucleotide substitutions per site, per year, respectively), indicating that these viruses exhibit fundamentally different evolutionary dynamics. The time to the most recent common ancestor of both IHNV and VHSV was consistent with the first reports of these pathogens in Italy. By combining sequence data with epidemiological information it was possible to identify different patterns of virus spread among trout farms, in which adjacent facilities can be infected by either genetically similar or different viruses, and farms located in different water catchments can be infected by identical strains. Overall, these findings highlight the importance of combining molecular and epidemiological information to identify the determinants of IHN and VHS spread, and to provide data that is central to future surveillance strategies and possibly control

Rabies and canine distemper virus epidemics in the red fox population of Northern Italy (2006–2010)

Since 2006 the red fox (Vulpes vulpes) population in north-eastern Italy has experienced an epidemic of canine distemper virus (CDV). Additionally, in 2008, after a thirteen-year absence from Italy, fox rabies was re-introduced in the Udine province at the national border with Slovenia. Disease intervention strategies are being developed and implemented to control rabies in this area and minimise risk to human health. Here we present empirical data and the epidemiological picture relating to these epidemics in the period 2006-2010. Of important significance for epidemiological studies of wild animals, basic mathematical models are developed to exploit information collected from the surveillance program on dead and/or living animals in order to assess the incidence of infection. These models are also used to estimate the rate of transmission of both diseases and the rate of vaccination, while correcting for a bias in early collection of CDV samples. We found that the rate of rabies transmission was roughly twice that of CDV, with an estimated effective contact between infected and susceptible fox leading to a new infection occurring once every 3 days for rabies, and once a week for CDV. We also inferred that during the early stage of the CDV epidemic, a bias in the monitoring protocol resulted in a positive sample being almost 10 times more likely to be collected than a negative sample. We estimated the rate of intake of oral vaccine at 0.006 per day, allowing us to estimate that roughly 68% of the foxes would be immunised. This was confirmed by field observations. Finally we discuss the implications for the eco-epidemiological dynamics of both epidemics in relation to control measures

Farmers’ Biosecurity Awareness in Small-Scale Alpine Dairy Farms and the Crucial Role of Veterinarians

Background: Biosecurity measures are essential to improve animal health and welfare, tackle antimicrobial resistance (AMR), minimize the burden of infectious diseases, and enhance the safety, security, and quality of sustainable agricultural products. However, the implementation of these measures in small mountain farms can be influenced by several factors, which probably differ from the main variables that affect intensive farming systems. Methods: This study investigated the awareness among farmers regarding the implementation of biosecurity measures at the local level in small dairy farms located in the Autonomous Province of Bolzano/Bozen (Italy). In order to determine to what extent these measures are in line with the recommendations, a questionnaire was conceptualized and sent by post to a representative group of local farmers. The initiative was voluntary and anonymous. Results: A total of 50 farmers responded to the questionnaire, resulting in a response rate of 27.5%. This study confirms that most of the farmers are familiar with biosecurity, and 38% of them know its comprehensive definition. The results indicate that 77% of respondents are willing to implement preventive measures to reduce the use of antimicrobials, and 76% of them acknowledge that they would follow the recommendations provided by veterinarians. In agreement with other studies, the role of the veterinarian as a person of trust among farmers is confirmed. Conclusions: Main strengths and areas of improvement have been identified. Additional data and effective tools are needed to better investigate drivers toward biosecurity and to implement impactful and practical measures for mountain farming

An Innovative Fluid Dynamic System to Model Inflammation in Human Skin Explants

Skin is a major administration route for drugs, and all transdermal formulations must be tested for their capability to overcome the cutaneous barrier. Therefore, developing highly reliable skin models is crucial for preclinical studies. The current in vitro models are unable to replicate the living skin in all its complexity; thus, to date, excised human skin is considered the gold standard for in vitro permeation studies. However, skin explants have a limited life span. In an attempt to overcome this problem, we used an innovative bioreactor that allowed us to achieve good structural and functional preservation in vitro of explanted human skin for up to 72 h. This device was then used to set up an in vitro inflammatory model by applying two distinct agents mimicking either exogenous or endogenous stimuli: i.e., dithranol, inducing the contact dermatitis phenotype, and the substance P, mimicking neurogenic inflammation. Our in vitro system proved to reproduce inflammatory events observed in vivo, such as vasodilation, increased number of macrophages and mast cells, and increased cytokine secretion. This bioreactor-based system may therefore be suitably and reliably used to simulate in vitro human skin inflammation and may be foreseen as a promising tool to test the efficacy of drugs and cosmetics

Risk factors linked to the onset of Nodular Gill Disease (NGD) in rainbow trout farms in North-eastern Italy

Nodular gill disease (NGD) is an emerging disease caused by amoebae affecting freshwater salmonids. It can cause high mortality and severe economic losses, in particular in rainbow trout (Oncorhynchus mykiss) farms. In Italy, most of the production of this species is found in the North-eastern regions. Therefore, an area in this part of the Country was selected to perform an epidemiological study with the aim of identifying the risk factors related to the onset of the disease. The survey was carried out between 2017 and 2019 and involved all the rainbow trout farms located in the Autonomous Province of Trento (North-eastern Italy). We administered a questionnaire to the 50 farmers, requesting information on the health status of the farms, their structural and management features, the applied biosecurity measures and the proximity and hydrological connection to other farms. Data were stored in a dedicated Access database, validated and analysed through statistical software STATA 12.1 and SAS 9.4. Univariate statistics were used to summarize the data characteristics. Afterwards, almost 70 variables were analysed through bivariate analysis (Chi-square or Fisher exact test for categorical variables, Wilcoxon/Mann-Whitney nonparametric test for two independent samples when quantitative variables were considered). A significant association (p-value<0.05) with NGD was detected for 7 variables. A logistic regression analysis was then performed with stepwise selection on variables that resulted significantly associated with NGD. The data analysis revealed that 42% of the farms experienced at least one episode of NGD in the previous 5 years. Furthermore, the reported cases increased during the study period, particularly in winter. The identified risk factors for the onset of NGD were the presence of other diseases in the farm in recent years (OR=17.496; 95% IC=2.746; 111.454), such as viral haemorrhagic septicaemia (VHS), infectious haematopoietic necrosis (IHN) and enteric redmouth disease (ERM), and having farms in the 5 km upstream (OR=24.679; 95% IC=2.885; 211.116). The results show that the disease is widespread among rainbow trout farms in the selected study area and underline the relevance of the exposure via water to amoebae and lack of biosecurity measures for the onset of the disease

First report of a fish kill episode caused by pyrethroids in Italian freshwater

Fish kills are events of strong emotional impact on the population because of the frequent suspicion that they can be the result of serious pollution accidents. As a matter of fact, they are often due to natural occurrences, such as low levels of dissolved oxygen in the water, but in many cases the causes remain unknown. Fish are particularly sensitive to pesticides and pyrethroids are reported to be the most ecotoxicologically active in the aquatic environment. Nevertheless, the reported cases of massive wild fish mortalities due to these toxicants are very few. This paper describes a fish kill episode occurred in the Padua Province (Veneto Region - North Eastern Italy) which involved several fish species and for which it was possible to identify the cause in the presence of pyrethroids in the water

Epidemiology of animal poisoning: An overview on the features and spatio-temporal distribution of the phenomenon in the north-eastern Italian regions

In the present paper we analyze and discuss about the records referring to animal poisonings and poisoned baits cases covering the period between 2007 and 2013 and submitted for diagnostic investigations to the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), which is the public veterinary health institute competent for the north eastern Italian regions. All data were gathered by a passive surveillance system based on voluntary reporting, which became mandatory in 2009 after a decree of the Italian Ministry of Health had come into force. This prohibited the use and detention of poisoned baits and ordered to selected institutions and professionals to carry out standardized surveys to assess suspect and/or confirmed reported cases; all the necessary anatomopathological and toxicological investigations to confirm the reported cases were then performed for free by public veterinary health institutes whenever a veterinarian diagnosis or clinical suspicion were provided. Totally, 1831 suspected animals poisoning and 698 cases of supposed poisoned baits recovery episodes were registered. 642/1831 (35.1%) animal poisoning cases were confirmed and the presence of toxic agents was verified in 292/698 baits (41.8%). The most severely affected territories were the ones with the highest level of urbanization and those most densely populated in the study area. Dogs and cats seemed to be greatly affected by poisoning cases and a characteristic seasonal trend was noticed, with an increase of episodes in late Winter/early Spring and in Autumn. Carbamate insecticides resulted to be the main cause for animal poisoning, while anticoagulants rodenticides played a primary role among toxicants found in poisoned baits. The presented results emphasize that malicious animal poisoning is a widespread problem in north-eastern Italy. The still relevant number of reported poisoning events caused by some banned pesticides poses the problem of identifying where these substances come from and brings to light the popular knowledge about the high toxicity of these compounds. Moreover, the noticeable increase of the number of episodes registered in 2009 pointed out how the above mentioned decree may have contributed to reveal a number of hidden cases which had not been investigated before, probably due to economic reasons related to the costs of toxicological analyses

Ex Vivo Evaluation of Ethosomes and Transethosomes Applied on Human Skin: A Comparative Study

In this study, the transdermal fate of vesicular nanosystems was investigated. Particularly, ethosomes based on phosphatidylcholine 0.9% w/w and transethosomes based on phosphatidylcholine 0.9 or 2.7% w/w plus polysorbate 80 0.3% w/w as an edge activator were prepared and characterized. The vesicle mean size, morphology and deformability were influenced by both phosphatidylcholine and polysorbate 80. Indeed, the mean diameters of ethosome were around 200 nm, while transethosome’s mean diameters were 146 or 350 nm in the case of phosphatidylcholine 0.9 or 2.7%, w/w, respectively. The highest deformability was achieved by transethosomes based on phosphatidylcholine 0.9%, w/w. The three types of vesicular nanosystems were applied on explanted human skin maintained in a bioreactor. Transmission electron microscopy demonstrated that all vesicles were able to enter the skin, keeping their structural integrity. Notably, the vesicle penetration capability was influenced by their physical-chemical features. Indeed, ethosomes reached keratinocytes and even the dermis, phosphatidylcholine 0.9% transethosomes were found in keratinocytes and phosphatidylcholine 2.7% transethosomes were found only in corneocytes of the outer layer. These findings open interesting perspectives for a differentiated application of these vesicles for transdermal drug delivery as a function of the cutaneous pathology to be addressed