Neuronal and astrocytic involvement in striped dolphins (Stenella coeruleoalba) with morbilliviral encephalitis.

Dolphin morbillivirus (DMV), a highly pathogenic agent, may cause peculiar, "brain-only" forms of infection (BOFDI), in which viral antigen and/or genome is found exclusively in the brain from striped dolphins (Stenella coeruleoalba). These BOFDIs show morphopathological similarities with subacute sclerosing panencephalitis and old dog encephalitis (ODE) in measles virus-infected patients and in canine distemper virus-infected dogs, respectively. The brain tissue from 3 BOFDI-affected striped dolphins was investigated by means of double labelling-indirect immunofluorescence (DL-IIF) and ultrastructurally, in order to characterize the DMV-targeted neuronal and non-neuronal cell populations, along with the associated submicroscopic findings. Viral colonization of calbindin-immunoreactive (IR) and nitric oxide synthase-IR neurons was detected in the cerebral parenchyma from the 3 DMV-infected dolphins under study, associated with nuclear (chromatin) and cytoplasmic (mitochondrial) ultrastructural changes. Furthermore, a limited viral targeting of brain astrocytes was found in these animals, all of which exhibited a prominent astrogliosis/astrocytosis. To the best of our knowledge, those herein reported should be the first submicroscopic pathology and neuropathogenetic data about BOFDI in striped dolphins. In this respect, the marked astrogliosis/astrocytosis and the low viral colonization of brain astrocytes in the 3 DMV-infected dolphins under investigation are of interest from the comparative pathology and viral neuropathogenesis standpoints, when compared with ODE-affected dogs, in whose brain a non-cytolytic, astrocyte-to-astrocyte infectious spread has been recently documented. Further studies aimed at characterizing the complex DMV-host interactions in BOFDI-affected striped dolphins are needed.

Western blot expression of 5-lipoxygenase in the brain from striped dolphins (stenella coeruleoalba) and bottlenose dolphins (tursiops truncatus) with or without encephalitis/meningo-encephalitis of infectious nature.

Dolphin Morbillivirus (DMV), Toxoplasma gondii and Brucella ceti are pathogens of major concern for wild cetaceans. Although a more or less severe encephalitis/meningo-encephalitis may occur in striped dolphins (Stenella coeruleoalba) and bottlenose dolphins (Tursiops truncatus) infected by the aforementioned agents, almost no information is available on the neuropathogenesis of brain lesions, including the neuronal and non-neuronal cells targeted during infection, along with the mechanisms underlying neurodegeneration. We analyzed 5-lipoxygenase (5-LOX) expression in the brain of 11 striped dolphins and 5 bottlenose dolphins, affected or not by encephalitic lesions of various degrees associated with DMV, T. gondii and B. ceti. All the 8 striped dolphins with encephalitis showed a more consistent 5-LOX expression than that observed in the 3 striped dolphins showing no morphologic evidence of brain lesions, with the most prominent band intensity being detected in a B. ceti-infected animal. Similar results were not obtained in T. gondii-infected vs T. gondii-uninfected bottlenose dolphins. Overall, the higher 5-LOX expression found in the brain of the 8 striped dolphins with infectious neuroinflammation is of interest, given that 5-LOX is a putative marker for neurodegeneration in human patients and in experimental animal models. Therefore, further investigation on this challenging issue is also needed in stranded cetaceans affected by central neuropathies.

5-Lipoxygenase and cyclooxygenase-2 in the lungs of pigs naturally affected by enzootic pneumonia and porcine pleuropneumonia.

Enzootic pneumonia by Mycoplasma hyopneumoniae and pleuropneumonia by Actinobacillus pleuropneumoniae are among the most common and economically relevant pulmonary diseases in swine herds. We herein investigated the activity and expression of 5-lipoxygenase (5-LOX) and cyclooxygenase-2 (COX-2) in healthy and diseased porcine lungs, by means of immunohistochemical, immunochemical and biochemical assays. Diseased lungs showed a significantly higher activity and expression of 5-LOX and COX-2 in a wide range of cell types, thus suggesting the likely involvement of both enzymes in the pathogenesis of bacterial porcine pneumonia. Consistently, increased enzyme activities were paralleled by increased leukotriene B(4) (LTB(4)), a 5-LOX product and prostaglandin E(2) (PGE(2)), a COX-2 product, content in diseased versus healthy lungs.

Intake of essential and non-essential elements from consumption of octopus, cuttlefish and squid.

Total concentrations of essential (Cu, Zn, Se and Cr) and non-essential (Hg, Cd, Pb and As) trace elements were measured in the flesh and hepatopancreas of Octopodidae (Eledone moschata, Eledone cirrhosa, Octopus salutii), Sepiidae (Sepia elegans, Sepia orbignyana) and Loliginidae (Illex coindeti, Loligo vulgaris) from the Mediterranean Sea. As expected, the hepatopancreas showed higher metal concentrations than flesh; the only exceptions were Hg and As, which were equally distributed in the two tissues. Regarding the edible portion, the highest toxic metal concentrations were in Octopodidae (Hg: 0.44, Cd: 0.49, Pb: 0.10 µg g(-1) wet weight) and Sepiidae (Hg: 0.27, Cd: 0.50, Pb: 0.12 µg g(-1) wet weight), while Loliginidae tended to accumulate less metal, especially Hg (Hg: 0.11, Cd: 0.30, Pb: 0.05 µg g(-1) wet weight). The other elements showed a heterogeneous distribution among the different cephalopod families. Loliginidae showed the highest Se concentrations (1.18 µg g(-1) wet weight), Octopodidae of Cu (37.37 µg g(-1) wet weight) and Zn (42.00 µg g(-1) wet weight) and Sepiidae of As (61.43 µg g(-1) wet weight), while Cr was uniformly distributed among the various families (0.38-0.43 µg g(-1) wet weight). In these seafoods, the concentrations of essential and non-essential elements were within the prescribed limits set by various authorities, except for Cu and As. Health risks posed by toxic elements to humans via dietary intake of these mollusks were assessed on the basis on Provisional Tolerable Weekly Intake (PTWI), while the estimated intakes of essential elements were compared to Dietary Reference Intakes (RDIs). A 70-g serving of these mollusks was shown to provide a large contribution to Cd intake (0.89 µg kg(-1) body weight), corresponding to 35.6% of PTWI. Concerning the essential elements, the consumption of these mollusks made an important contribution to daily dietary intake of Se, Cu and Zn.

5-lipoxygenase and cyclooxygenase-2 in porcine parasitic bronchopneumonia: immunohistochemical and biochemical investigations.

Eicosanoids are products of arachidonic acid metabolism and have numerous biological roles. The present study aimed to investigate the role of 5-lipoxygenase (5-LOX)- and cyclooxygenase-2 (COX-2)- dependent enzymatic pathways in the pathogenesis of porcine parasitic bronchopneumonia caused by Metastrongylus spp. Pulmonary tissue samples from healthy control and parasitized pigs were processed for histopathological, immunohistochemical and biochemical investigations. In control animals, immunohistochemistry demonstrated that 5-LOX and COX-2 expression was almost exclusively limited to the bronchiolar epithelial cells. Parasitized pigs had greater 5-LOX- and COX-2- specific immunoreactivity, involving a wide range of cell types within foci of granulomatous and eosinophilic bronchopneumonia. Biochemical investigations demonstrated the presence of 5-LOX (and the related product Leukotriene B(4)) and COX-2 (and the related product prostaglandin E(2); PGE(2)) in all tissues under study. COX-2 activity and PGE(2) concentration were significantly higher in diseased lungs compared with normal healthy controls. These findings demonstrate that 5-LOX and COX-2 are differentially expressed in normal versus lungworm-infected lungs and therefore suggest that both biochemical pathways are likely to be involved in the pathogenesis of porcine parasitic bronchopneumonia.

Cadmium and mercury in cephalopod molluscs: estimated weekly intake.

Cadmium and mercury concentrations were measured in the flesh and hepatopancreas of different species of cephalopod molluscs (European squid, common octopus, curled octopus, horned octopus, pink cuttlefish, common cuttlefish) in order to establish whether the concentrations exceeded the maximum levels fixed by the European Commission. In hepatopancreas, the levels of cadmium were substantially higher than those in flesh (flesh 0.11-0.87 microg g(-1) wet weight, hepatopancreas 2.16-9.39 microg g(-1) wet weight), whilst the levels of mercury (flesh 0.13-0.55 microg g(-1) wet weight, hepatopancreas 0.23-0.79 microg g(-1)) were approximately double those in flesh. Concentrations exceeding the maximum permitted limit of cadmium were found in 39.8 and 41.0% of common octopus and pink cuttlefish flesh, respectively. For mercury, concentrations above the limit were found only in octopuses, and precisely in 36.8, 50.0 and 20.0% of flesh samples of common, curled and horned octopus, respectively. In the hepatopancreas, concentrations of cadmium and mercury were above the proposed limits in all the samples examined. The estimated weekly intake of between 0.09 and 0.49 microg kg(-1) body weight for cadmium and between 0.05 and 0.24 microg kg(-1) body weight for mercury made only a small contribution to the provisional tolerable weekly intake (cadmium 1.3-7.0%, mercury 1.0-4.8%) set by the WHO.

Total mercury and methylmercury content in edible fish from the Mediterranean Sea.

The objective of this study was to determine the current levels of total mercury and methylmercury in the muscle tissues of different fish species caught in the Mediterranean Sea to ascertain whether these concentrations exceed the maximum level stipulated by the European Commission Decision. Total mercury concentrations in the muscles of skates ranged from 0.18 to 1.85 mg/kg (wet weight) (average, 1.00 mg/kg) while levels of 0.11 to 1.92 mg/kg (wet weight) (average, 0.70 mg/kg) and 0.21 to 1.74 mg/kg (wet weight) (average, 0.70 mg/kg) were recorded for blue whiting and red mullet, respectively. For 66.7% of long nose skate samples, 61.4% of thornback ray samples, 42.8% of winter skate samples, and 38% of starry ray samples, the total mercury concentrations exceeded the prescribed legal limit (1.0 mg/kg [wet weight]). Concentrations exceeding the maximum total mercury level stipulated by the European Commission Decision (0.5 mg/kg [wet weight]) were observed in 63.6 and 40% of blue whiting and striped mullet samples, respectively. Mercury was present in the different species almost completely in the methylated form at 55 to 100%. Weekly intakes were estimated and compared with the provisional tolerable weekly intake recommended by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives.

Survey of total mercury and methylmercury levels in edible fish from the Adriatic Sea.

Total mercury and methylmercury concentrations were measured in the muscle tissue of different fish species from the Adriatic Sea to ascertain whether the concentrations exceeded the maximum level fixed by the European Commission. Large species-dependent variability was observed. The highest total mercury mean concentrations were in benthic (0.20-0.76 microg g(-1) wet wt) and demersal fish (0.22-0.73 microg g(-1) wet wt), while pelagic species showed the lowest levels (0.09-0.23 microg g(-1) wet wt). In 15% of frost fish, in 42% of skate and in 30% of angler fish samples total mercury concentrations exceeded the maximum level fixed by the European Commission (Hg = 1 microg g(-1) wet wt); for the species for which the maximum level was set to 0.5 microg g(-1) wet wt, concentrations exceeding the prescribed legal limit were observed in 6.4% of bokkem, in 6.6% of pandora, in 20% of megrin, in 12.5% of four-spotted megrim, in 16% of striped mullet, in 5.0% of forkbeard and in 5.3% of picarel samples. In all the different species, mercury was present almost completely in the methylated form, with mean percentages between 70 and 100%. Weekly intake was estimated and compared with the provisional tolerable weekly intake recommended by the FAO/WHO Expert Committee on Food Additives. A high exposure was associated with the consumption of only skates, frost fish and angler fish, thought the consumption of the other species, such as, megrim, four spotted megrim, red fish striped mullet and forkbeard, resulted in a weekly intake slightly below the established provisional tolerable weekly intake.

Total and methylmercury residues in cartilaginous fish from Mediterranean Sea.

Concentrations of mercury and methylmercury residues were determined in the muscle tissue of three cartilaginous fishes Chimaera monstrosa (ghostshark), Torpedo nobiliana (electric ray) and Myliobatis aquila (eagle ray) from the Mediterranean Sea. The highest mean levels of total mercury were detected in ghostshark (3.14 mg/kg wet weight), followed by electric ray (2.42 mg/kg wet weight), and eagle ray (0.83 mg/kg wet weight). Such a variability is influenced by quite a number of factors, among which size, fish ecology and feeding habits. The percentages of the methylated form to total mercury ranged from a minimum of 72% in eagle ray to a maximum of 83% in ghostshark. Relationship between specimen size and mercury and methylmercury concentrations were found in ghostshark species.

Total mercury in muscle of benthic and pelagic fish from the South Adriatic Sea (Italy).

Total mercury concentrations were measured from the muscle of different kinds of fish: yellow gurnard (Trigla lucerna), red gurnard (Aspitrigla cuculus) red fish (Helicolenus dactylopterus), skate spp. (Raje spp.), goldline (Sarpa salpa), atlantic bonito (Sarda sarda), mackerel (Scomber scombrus), chub mackerel (Scomber japonicus) caught in the South Adriatic Sea (south Italy) in the period June/August 1995. The highest total mercury levels were found in the benthic marine organisms and particularly in skates (Raje spp.) whose values ranged from 0.05 to 2.65 mg/kg wet wt with a mean value of 1.02 mg/kg wet wt. As for pelagic species, the highest mean levels were observed in Atlantic bonito (0.34 mg/kg wet wt), while in goldline the mean content of total mercury was the lowest (0.07 mg/kg wet wt). According to the rules in force (Official Journal of the European Communities 1994) 53% of skate and Atlantic bonito samples showed concentrations exceeding the peak value of 1 mg/kg, while for the other species, only 28% of samples exceeded the peak value fixed at 0.5 mg/kg. Correlations between total mercury concentration and specimen weight were evident in all examined species except for goldline and skates.