Intentional poisoning represents a serious risk to domestic and wild animals, and it can be an environmental and human health issue as well . This paper is a retrospective study, which covers a decade, based on animal poisoning cases and poisoned baits that were submitted for diagnostic examinations to the Veterinary Medical Research Institute for Piedmont, Liguria and the Aosta Valley (IZS-PLVA) in Liguria region. All data were collected through a passive surveillance system introduced in Italy by a decree of the Ministry of Health in January 2009. 43.2% of the animal poisoning cases were confirmed by toxicological analysis, whereas toxic agents were detected in 31.1% of the baits. The most affected animal species were dogs and cats, followed by synanthropic birds,. Only 4% of the total poisoning events analysed involved wild animals and cases of livestock poisoning were minimal. An increased number of cases in January, March, April and August was noticed, but no seasonal trend was detected. The most affected areas were the ones with the highest level of urbanization and population density. The major cause of the poisonings and the most common substances detected in the examined baits were anticoagulants whereas cholinesterase inhibitors, organochlorine pesticides and carbamates were detected in a minor number of cases. This study raises concerns about deliberate animal poisoning in ligurian region and highlights the necessity to fight this phenomenon as it endangers animals, humans and environment.

Purpose: Pathogens of the genera Anaplasma, Borrelia and Rickettsia, and Tick-borne Encephalitis virus (TBEV) may be associated to various tick species which are hematophagous ectoparasites potentially hosted by any terrestrial vertebrate, humans included. This peculiarity makes ticks potential horizontal vectors of zoonotic importance. Liguria is characterized by mountains and an intense urbanization of the terminal part of the alluvial plains. The 12% of its surface is a protected natural area. The aim of our study is to provide an overview of the infection rate of tick-borne agents by analyzing ticks collected between 2016 and 2017 from wild mammals in Liguria. Methods & Materials: More than 200 ticks (124 Ixodes, 68 Dermacentor, 34 Haemaphysalis and 9 Riphicephalus spp.) were collected from 49 roe deer (Capreolus capreolus), 41 wild boars (Sus scrofa), 7 chamois (Rupicapra rupicapra), 2 fallow deer (Dama dama) and 1 marten (Martes foina) and stored in 70% ethanol. After ticks identification and total nucleic acid extraction, simplex PCR assays to detect Anaplasma, Borrelia, Rickettsia spp. and TBE virus were performed. By project's design, all ticks belonging to a given species from the same animal and collection date were pooled and treated as a single sample. Results: PCR results revealed 36 ticks positive for Rickettsia spp. and 3 for Anaplasma spp.; all samples were negative for Borrelia spp. and TBEV. Rickettsia spp. were found in 32 Ixodes ricinus and 4 Dermacentor marginatus ticks, while Anaplasma spp. were related to 2 Haemaphysalis punctata and 1 Ixodes ricinus (co-infected with Rickettsia). Regarding the host species, the 15% of wild boars and 61% of roe deer sampled were carrying ticks infected with Rickettsia, while the 29% of chamois ticks infected with Anaplasma. Conclusion: Our study confirms the presence of Rickettsia and Anaplasma spp. in ticks in Liguria, giving proof of their circulation in wild mammals. This panorama suggests the importance of taking forward the health monitoring in order to ensure the early detection of potential zoonotic infections in this area.

Information regarding chemical pollutant levels in farmed fish and shellfish, along with the risks associated with their consumption is still scarce. This study was designed to assess levels of exposure to 21 trace elements in fish (Dicentrarchus labrax), mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected from aquaculture marine ecosystems of the northwestern Mediterranean Sea. Metal concentrations showed great variability in the three species; the highest values of the nonessential elements As and Cd were found in oysters while the highest levels of Al, Pb and V were found in mussels. The essential elements Cu, Mn and Zn were highest in oysters, but Fe, Cr, Ni, Se, Co and Mo levels were highest in mussels. Fish had the lowest concentrations for all trace elements, which were at least one order of magnitude lower than in bivalves. The rare earth elements cerium and lanthanum were found at higher levels in mussels than in oysters, but undetectable in fish. The maximum values set by European regulations for Hg, Cd and Pb were never exceeded in the examined samples. However, comparing the estimated human daily intakes (EHDIs) with the suggested tolerable copper and zinc intakes suggested a potential risk for frequent consumers of oysters. Similarly, people who consume high quantities of mussels could be exposed to concentrations of Al that exceed the proposed TWI (tolerable weekly intake).

The paramyxean protistan parasites Marteilia refringens and M. maurini, two Marteilia species detected in Europe, infect several bivalve mollusc species of economic interest. Marteiliosis, caused by infection with M. refringens, is included in the World Organization for Animal Health (OIE) list of diseases (OIE Aquatic Animal Health Code 2014) and is listed as a nonexotic pathogen under European Union legislation (2006/88/EC). As mandated by Italian Legislative Decree 148/2008, an annual sampling program is in place to monitor the health status of the Mediterranean mussel Mytilus galloprovincialis farmed in the Gulf of La Spezia (northwest Italy), test for the presence of the parasite in mussels, and evaluate abnormal mortality as an indicator of disease presence. During the four-year period 2010-2013, 4460 samples of M. galloprovincialis were collected from six farming areas and analyzed by cytological methods for the detection of M. refringens. Suspected positive diagnosis was confirmed by PCR assay performed according to the Manual of Diagnostic Tests for Aquatic Animals (OIE, 2012). Although infection rates generally decreased between 2010 and 2013, samples from all six areas tested positive for M. refringens, with an overall positivity rate of 4.05%.

Mycobacterium spp. and Photobacterium damselae subsp. piscicida are recognized as the most frequent causative agents of granulomatous lesions in fish. Although frequent episodes of mycobacterial infections have been reported in wild fish worldwide, only sporadic cases have been documented to date in Italy. To investigate for the presence of lesions referable to mycobacteriosis and to identify the mycobacterial species involved, a total of 159 wild mullets were fished from the eastern coast of the Ligurian Sea, killed and necropsied. Liver and spleen samples were collected from all fish for histopathological and microbiological analyses. Molecular investigations for identification of Photobacterium damselae subsp. piscicida were performed. Gross examination revealed granulomatous lesions in one animal; microscopically, 42.14% of fish displayed granulomas with various histological features, 19.50% resulted positive at Ziehl-Neelsen staining, and were confirmed as mycobacterial lesions by culture. The identified colonies were characterized as M. fortuitum, M. abscessus, M. flavescens, M. chelonae, M. septicum and M. nonchromogenicum. In all, 35% of animals resulted positive for Photobacterium damselae subsp. piscicida. These data suggest widespread mycobacterial infection also by Photobacterium damselae subsp. piscicida infections in wild fish. Moreover, the pathogenicity of some mycobacterial species, previously considered as saprophytic, was demonstrated.

Vibrio parahaemolyticus is a common microflora of marine and estuarine waters known to be associated with gastroenteritis due to consumption of raw seafood, cooked food contaminated through direct contact (cross contamination) or improperly cooked. The aim of this study was to evaluate the effectiveness of two different heat processes in reducing microbial load of V. parahaemolyticus. Data show a 4 Log10/g reduction of bacterial load after 3,30 and 15 minutes at 95 °C.

AIMS:The aim of this study was to investigate the presence of Vibrio vulnificus and potentially pathogenic strains of Vibrio parahaemolyticus in mullets collected from estuarine environment in Italy.METHODS AND RESULTS:Two hundred and ninety-five mullets were analysed by culture using the selective medium thiosulfate citrate bile salt sucrose agar, during a monitoring period of 2 years (2008-2009). Presumptive Vibrio colonies were initially identified by using biochemical tests, and strains identified as V. parahaemolyticus and V. vulnificus were subsequently examined by PCR for the presence of species-specific and virulence genes (toxR, trh, tdh and vvh). V. parahaemolyticus was found in 55% (162/295) of fishes and V. vulnificus in 1% (3/295) with a higher presence in summer months. The trh+/tdh- strains were detected in 16% (47/295) of samples and only one strain resulted trh+/tdh+. One of the V. parahaemolyticus trh+ strains isolated belonged to the O1:KUT (K untypeable), a serotype recently associated to gastroenteritis in Italy.CONCLUSIONS:This is the first report demonstrating a high percentage of potential pathogenic V. parahaemolyticus trh+ strains in estuarine fishes of the Mediterranean area.SIGNIFICANCE AND IMPACT OF THE STUDY:These findings indicate the potential human health risk associated with the presence of pathogenic Vibrio spp. in wild fishes.

Aims:The purpose of this work was to verify whether E. coli is a good indicator of viral contamination in mussels and Adenovirus could represent a better alternative as indicator organism of viral presence to guarantee consumer health protection.Methods and Results:Eighty samples of mussels from La Spezia Gulf were analysed for E. coli, Salmonella, Adenovirus, Norovirus and hepatitis A virus with cultural and biomolecular tests. The results of bacterial parameters showed E. coli within the law's limits and the absence of Salmonella. Twelve samples were positive for Adenovirus presence, one for Norovirus genogroup II and two for hepatitis A virus. None of these positive mussels was found to be contaminated with more than one virus at the same time.Conclusion:This study showed that there was not a direct correlation between the presence of human pathogenic viruses and bacterial indicators.Significance and Impact of the Study:Both E. coli and Adenovirus cannot be considered valid substitutes for the direct research of human pathogenic viruses in mussels. To improve consumer health protection, the European Commission will provide standardized methods for Norovirus and hepatitis A virus detection as soon as possible.

In this study it has been evaluated the correlation between the biological test on mice and the assay on cultured cells for the determination of okadaic add, on artificially contaminated mussels. Particularly the study has been made in vitro on two lines of cells KB (Human oral epidermoid carcinoma) and H9c2 (Rat heart myoblast) verifying the sensibility of two models and the limit of revealability in various protocol of analysis. The obtained results show a higher sensibility of the KB line compared to the H9c2 line in the studied conditions. Only for the KB line it has been observed a limit of the same revealability of the biological test (0.5 μg/g). The correlation between biological test and method on the KB line resulted significative: correlation index (Ρ 2 = 0.606) and Pearson's coefficient equal to -0.78.

The Italian national law (D.Lvo n. 530 del 30/12/92), sets the control of viruses in mussel, subordinating it to the presence of routine methods and virological rules.In this study different methods have been evaluated for enterovirus recovery. The examined methods resulted to be very reliable and simple (percent recovery 98-100%).

The Italian national law (D.Lvo n. 530 del 30/ 12/92), sets the control of viruses in mussel, subordinating it to the presence of routine methods and virological rules. In this study different methods have been evaluated for enterovirus recovery. The examined methods resulted to be very reliable and simple (percent recovery 98-100%).