Do Organochlorine Contaminants Modulate the Parasitic Infection Degree in Mediterranean Trout (Salmo trutta)?

We investigated the occurrence of organochlorine pollutants (OCs) in the muscle of brown trout and evaluated their potential modulation of parasite infection. The toxicological risk for consumer health was assessed, too. Trout were collected from the Sila National Park (Calabria region, South of Italy). The highest concentrations emerged for the sum of the 6 non-dioxin-like (ndl) indicator polychlorinated biphenyls (Σ6ndl-PCBs), followed by the 1,1,1-trichloro-2,2-di(4-chlorophenyl)-ethane (DDT), dioxin-like PCBs, hexachlorobenzene (HCB), and dieldrin. Measured on lipid weight (LW), the mean value of Σ6ndl-PCBs amounted to 201.9 ng g-1, that of ΣDDTs (the sum of DDT-related compounds) to 100.2 ng g-1, with the major contribution of the DDT-metabolite p,p'-DDE which was detected in all sample units (97.6 ng g-1 on average). Among dioxin-like congeners, PCB 118 showed the highest mean concentration (21.96 ng g-1 LW) and was detected in all sample units. Regression analysis of intestinal parasites on OC concentration was performed, controlling for two potential confounding factors, namely sex and sexual stage. The results evidenced the existence of interactions between the dual stressors in the host-parasite system in the wild. A negative and statistically significant correlation was estimated, suggesting that OCs may decrease parasite infection degree. Regarding the toxicological risk evaluation, OC concentrations were consistently below the current European Maximum Residue Limits.

Seroprevalence and phylogenetic characterization of hepatitis E virus in pig farms in Southern Italy.

Hepatitis E virus (HEV) infection is an emerging public health problem in industrialized countries. The infection is associated with waterborne epidemics and transmitted via faecal-oral route. Zoonotic cases of HEV in humans have increased in Europe, and HEV genotype 3 (HEV-3) is the most frequent among humans and animals. Nevertheless, HEV surveillance in the Italian pig farming industry is patchy. Here, HEV prevalence in pig farms located in the Calabria region in Southern Italy was investigated. A total of 692 serum samples were collected from 26 farms and tested for anti-HEV IgG antibody detection. The percentage of HEV-seropositive pigs was 56.8 %. Small farm size, farrow-to-finishing production, and infrequent cleaning procedures were associated with higher HEV seroprevalence. In 12 of the HEV-seropositive farms, 67 faecal samples were collected and 10 of these (10.6 %) tested positive for HEV RNA. Seven of 10 viral RNA sequences were genotyped for phylogenetic analysis, five of which belonged to subtype HEV-3f and two to subtype HEV-3e. The high HEV seroprevalence and the circulation of HEV-3 strains among domestic pigs in the Calabria region pose a risk for the zoonotic transmission of HEV from pigs to occupational exposed workers.

Presence of Mycobacterium bovis in slaughterhouses and risks for workers.

An investigation was carried out to detect the presence of Mycobacterium bovis in slaughterhouses where intradermal tuberculin test positive cattle were slaughtered, and to evaluate the risk of contamination by M. bovis among exposed slaughterhouse workers. Swabs were taken from the carcasses of slaughtered animals showing autoptic signs of non-generalized forms of tuberculosis, thus authorized for free consumption. Swabs were also taken from the hands and clothes of the staff employed in the butchery production line. Environmental samplings were conducted on the slaughterhouse air using filters and air aspiration devices, and on water used to wash the carcasses after slaughter. Samples from the carcasses of healthy animals were also taken on a following slaughtering session. The swabs were analysed by means of Polymerase Chain Reaction for the detection of mycobacteria. M. bovis was detected on meats, on the hands of one worker, and in the washing water. The results obtained from this study confirm that workers are highly exposed to infection by zoonotic tuberculosis, and that cleaning procedures were ineffective in our setting.

Foodborne Pathogen Assessment in Raw Milk Cheeses.

General hygienic parameters and selected foodborne pathogens in raw milk cheeses at the retail level were evaluated. A total of 245 raw milk cheese samples were analysed for total bacterial count, Enterobacteriaceae, E. coli, Salmonella spp., Listeria monocytogenes, coagulase-positive Staphylococci, and staphylococcal enterotoxin. Results showed only 3 samples that were not compliant with European rules on staphylococcal enterotoxin, but coagulase-positive Staphylococci were evidenced in all samples tested. Salmonella spp. and Listeria monocytogenes were never detected whereas E. coli was evidenced in 20 samples. Results suggest a need for improvement of good manufacturing practice and milking operation.

Molecular detection and characterization of carnivore parvoviruses in free-ranging Eurasian otters (Lutra lutra) in southern Italy.

The most important Italian population of the Eurasian otter (Lutra lutra) occurs in the southern part of the peninsula with two isolated sub-populations of about 250 adult individuals. The Eurasian otter is considered to be near threatened and it is a fully protected species. The aims of this study were to investigate for the first time the occurrence and characterize the parvoviruses included in the species Carnivore protoparvovirus 1 in seven carcasses of road-killed Eurasian otters from the southern Italy. Carnivore protoparvovirus 1 are responsible for acute gastroenteritis and leukopenia in pets and free-ranging carnivores. Initial screening of tissue samples by real-time PCR revealed CPV/FPV DNA in tissue samples of five Eurasian otters; three of them, showed co-infections by both CPV and FPV. Among the five positive Eurasian otters, we successfully obtained six DNA sequences from four individuals including two CPV-2a, one CPV-2b, one CPV-2c, and two FPV sequences. Comparison of these sequences with 250 VP2 gene sequences deposited in the GenBank database, showed 10 nt differences resulting in two synonymous and eight non-synonymous substitutions. On the basis of these results, two sequences here found were characterized as new CPV-2a, one was characterized as new CPV-2b variant, and one was characterized as FPV-like mutant. The last two sequences belong to a FPV and CPV-2c strain respectively. Carnivore protoparvovirus 1 is reported for the first time in the Eurasian otter showing high infection value in southern Italy. Occurrence of this infection should be studied further to understand its possible pathogenicity and virulence to the fragile and isolate Eurasian otter population which live in southern Italy.

Quantitative determination by screening ELISA and HPLC-MS/MS of microcystins LR, LY, LA, YR, RR, LF, LW, and nodularin in the water of Occhito lake and crops.

The occurrence of harmful cyanobacterial blooms in surface waters is often accompanied by the production of a variety of cyanotoxins, and these toxins are designed to target in humans specific organs on which they act. When introduced into the soil ecosystem by spray irrigation of crops, they may affect the same molecular pathways in plants having identical or similar target organs, tissues, cells, or biomolecules. There are also several indications that terrestrial plants, including crops, can bioaccumulate cyanotoxins and present, therefore, potential health hazards for humans. During this project, for monitoring purposes, water samples were collected from lake Occhito, in which there was an algal bloom (Planktothrix rubescens) in 2009, and from three tanks which acted as hydraulic junctions. In addition, crop samples irrigated with water from the three tanks mentioned above were also picked. Finally, the characterization of principal cyanobacteria was performed, to determine the presence of cyanotoxins such as microcystins and validate a method of screening ELISA for the determination of microcystins in vegetable samples and a confirmatory method by HPLC-ESI-MS/MS. Graphical abstract Occhito lake (left), microcystin LR (center), Tomato field in Foggia (right); figures below: ELISA (left), HPLC-MS/MS (right).

Does the clam Ensis siliqua provide useful information about contamination by polychlorinated biphenyls and organochlorine pesticides beyond that of mussel Mytilus galloprovincialis?

Several polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) investigated in soft tissues of the frequently monitored Mytilus galloprovincialis were compared to those of Ensis siliqua, a highly dispersed and economically important bivalve species, though rarely investigated. Overall PCBs had higher concentrations than OCPs in both species with a prevalence of tri- tetra-and penta-chlorinated biphenyls in E. siliqua and a prevalence of hexa- hepta and octa-chlorinated biphenyls in M. galloprovincialis. E. siliqua emerges as a suitable complement to mussels for monitoring PCBs and OCPs pollution.

Polychlorinated biphenyls and organochlorine pesticides in European eel (Anguilla anguilla) from the Garigliano River (Campania region, Italy).

Measuring organochlorine compounds in muscle tissue of European eels from the Garigliano River in Campania (Italy), overall PCBs emerged as the most abundant pollutants, followed by DDTs, Dieldrin and HCB. Target PCBs, IUPAC nos. 118, 138, 153 and 180, were the dominant congeners accounting for 64.2% of total PCBs. Among OCPs, p,p'-DDE was detected in all eels, always with higher concentration levels than other OCPs; p,p'-DDT was frequently detected, about 93.3% of the sample. The high and statistically significant correlations between concentration and length as well as weight of eels suggest that the organochlorine compounds concentrations tend to increase with the size and consequently with the age. Concentrations of DDTs and PCBs detected were similar to those reported in studies relative to France, UK and Sweden. As regards toxicological risk for human health, in general OCPs residual levels were below the limits established for fish and aquatic products. Conversely, the concentrations of PCBs exceeded the limit set by the EU for terrestrial foods. Our results imply that OCPs and PCBs are still important persistent chemical contaminants in Campania freshwaters, although their manufacture and use are banned or highly restricted.

Polychlorinated biphenyls and organochlorine pesticides in seafood from the Gulf of Naples (Italy).

Seven target polychlorinated biphenyls (PCBs; IUPAC nos. 28, 52, 101, 118, 138, 153, and 180) and the organochlorine pesticides (OCPs) hexachlorobenzene (HCB) and dichlorodiphenyltrichloroethane (DDT) and its related metabolites (p,p'-DDT, p,p'-DDE, and p,p'-DDD) were quantified in edible tissues from seven marine species (European hake, red mullet, blue whiting, Atlantic mackerel, blue and red shrimp, European flying squid, and Mediterranean mussel) from the Gulf of Naples in the southern Tyrrhenian Sea (Italy). PCBs 118, 138, and 153 were the dominant congeners in all the species examined. The concentrations of all PCBs (from not detectable to 15,427 ng g(-1) fat weight) exceeded those of all the DDTs (from not detectable to 1,769 ng g(-1) fat weight) and HCB (not detectable to 150.60 ng g(-1) fat weight) in the samples analyzed. The OCP concentrations were below the maximum residue limits established for fish and aquatic products by the Decreto Ministerale 13 May 2005 in all the samples analyzed; therefore the OCPs in the southern Tyrrhenian Sea species are unlikely to be a significant health hazard. Conversely, the mean concentrations of PCBs exceeded (greatly in some cases) the current limits (200 ng(-1) fat weight) set by the European Union for terrestrial foods. Although the manufacture and use of PCBs are banned or highly restricted, these compounds still are important persistent chemical contaminants in the Gulf of Naples.