Survival of Escherichia coli O157:H7 during the manufacture and ripening of Cacioricotta goat cheese.

The aim of this study was to assess the growth and survival of Escherichia coli O157:H7 during the manufacturing and ripening of Cacioricotta goat cheese. Goat milk was artificially contaminated with E. coli O157:H7 and the bacterial load was monitored from production up to 90 days of ripening. Goat milk was inoculated with 102 cfu ml-1 of E. coli O157:H7 and the bacterial count of the curd at time zero was 2.31 log10 cfu g-1. During the first day of ripening, the bacterial load has increased to 5.73 log10 cfu g-1 to more than 6.20 log10 cfu g-1 during the first week. The bacterial load remained constant up to 28 days and then slightly decreased until the end of ripening, with values of aw and pH of 0.88 and 5.41 respectively. The results of this study highlighted that E. coli O157:H7 is able to survive the manufacturing process and they suggest that the 90-day period of ripening alone is insufficient to remove E. coli O157:H7 in contaminated Cacioricotta goat cheese. Moreover, these results support the assumption that the presence of a low contamination of milk with E. coli O157:H7 could represent a potential source of infection and a threat to consumers.

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.

Levels and congener profiles of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in sheep milk from an industrialised area of Sardinia, Italy.

Concentrations of 7 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs) and 22 polychlorinated biphenyls (PCBs), including 12 dioxin like-PCBs (non- and mono-ortho PCBs) were measured in 80 sheep milk samples from farms located in an industrialized area of Sardinia, Italy. PCDDs and PCDFs mean concentrations were 2.45 and 3.69 pgg(-1) fat basis, respectively. The mean dl-PCB concentration was 2.01 ngg(-1) fat basis, while cumulative ndl-PCB levels ranged from 1.02 to 20.42, with a mean of 4.92 ngg(-1) fat. The results expressed in pg WHO-TEQ/g fat showed that contamination level of milk was below the limit values for human consumption established by EC legislation. In the same way, all the investigated milk exhibited PCDD/Fs concentrations below EU action levels, while dl-PCBs concentrations exceeded the action level of 2.0 pg WHO-TEQ/g fat. These findings point to the need to continue to conduct general monitoring programmes, including also milk samples from areas not close to the contaminant-emitting industries, in order to better evaluate the impact of industrial activities on surrounding environment.

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.

Occurrence of heavy metals (Hg, Cd, and Pb) and polychlorinated biphenyls in salted anchovies.

The most popular brands of salted anchovies (Engraulis encrasicolus) from the Mediterranean Sea and Atlantic Ocean were purchased from several Italian supermarkets and grocery stores. Heavy metal (Hg, Cd, and Pb) and polychlorinated biphenyl (PCB) levels were determined and assessed by comparing the concentrations in these samples with the maximum permissible limits set by the European Union (Reg EC 629/2008 and Reg EC 1881/2006 [Off. J. Eur. Union L 173:3-9 and 364:5-24, respectively]). The Hg and Cd levels were higher than those of Pb in all samples examined. For Hg and Pb, the concentrations recorded in this study were below the authorized limits, while an appreciable percentage of samples from both locations (Mediterranean Sea, 35%, and Atlantic Ocean, 25%) showed Cd levels exceeding the threshold recommended for human consumption. Concerning PCBs, the results of principal component analysis showed that samples from the two different marine areas appeared to be discriminate, with Mediterranean anchovies more contaminated than the others, in spite of their lower lipid content. However, anchovy samples from both locations had dioxinlike-PCB levels (Mediterranean Sea, 0.011 pg World Health Organization toxic equivalency [WHO-TEQ] g(-1), wet weight, and Atlantic Ocean, 0.007 pg WHO-TEQ g(-1), wet weight) that were below the WHO-TEQ maximum concentration set by European regulation. The results of this study will help in generating data needed for the assessment of heavy metal and PCB intake from this food.

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.

Polychlorinated biphenyl and organochlorine pesticide contamination signatures in deep-sea fish from the Mediterranean Sea.

Polychlorinated biphenyl (PCB) and OCP concentrations were determined in the livers of two deep-sea fish species, roughsnout grenadier and hollowsnout grenadier, from the Adriatic Sea. In both species, contaminant concentrations were in the following order: PCBs>DDTs>>HCB. Contaminant load was higher in roughsnout grenadier (PCB: 12,327ngg(-1); DDTs: 5357ngg(-1); HCB: 13.1ngg(-1)) than in hollowsnout grenadier (PCB: 1234ngg(-1); DDTs: 763ngg(-1); HCB 6.3ngg(-1)). PCB patterns were dominated by higher chlorinated congeners (hexa-CBs: 50.3-52.1%, hepta-CBs: 29.6-35.5%, penta-CBs: 8.0-11.1% and octa-CBs :5.2-5.4%). PCBs 138, 153 180 and 187 were the most abundant. Regarding the DDT pattern, p,p'-DDE was prevalent in both species (roughsnout grenadier: 99.7%, hollowsnout grenadier: 90%), suggesting no recent DDT input. In both species, the total 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalent (TEQ) concentrations (roughsnout grenadier: mean 43.77pg/g, hollowsnout grenadier: mean 20.49pg/g), calculated from non- and mono-ortho PCBs, reached those encountered in marine organisms at higher levels in the trophic chain.

Intake of essential minerals and metals via consumption of seafood from the Mediterranean Sea.

Edible marine species (fish and cephalopod molluscs) from the Mediterranean Sea were analyzed for their metal content (Hg, Cd, Pb, Cr, Cu, Zn, and Ni). Human health risks posed by these elements via dietary intake of seafood were assessed based on the provisional tolerable weekly intake, reference dose, and recommended dietary allowances. Metal concentrations varied widely among the different organisms, indicating species-specific accumulation. On a wet weight basis, the maximum concentrations of Hg were found in fish (1.56 microg g(-1)), and the maximum concentrations of cadmium were found in cephalopod molluscs (0.82 microg g(-1)), whereas for Pb the concentrations were generally low (fish, 0.01 to 1.18 microg g(-1); cephalopod molluscs, 0.03 to 0.09 microg g(-1)). For the essential metals, cephalopods had higher concentrations (Cr, 0.40 microg g(-1); Zn, 33.03 microg g(-1); Cu, 23.77 microg g(-1); Ni, 2.12 microg g(-1)) than did fish (Cr, 0.17 microg g(-1); Zn, 8.43 microg g(-1); Cu, 1.35 microg g(-1); Ni, 1.13 microg g(-1)). The estimated weekly intake of Cd and Pb indicated increased health risks through the consumption of various seafoods. Conversely, a health risk was ascribed to the intake of Hg from consumption of certain fish, such as albacore (10.92 microg kg(-1) body weight) and thornback ray (5.25 microg kg(-1) body weight). Concerning the essential metals, cephalopod mollusc consumption made an important contribution to daily dietary intake of Cu, Zn, and Ni.

Potential human health risks from metals (Hg, Cd, and Pb) and polychlorinated biphenyls (PCBs) via seafood consumption: estimation of target hazard quotients (THQs) and toxic equivalents (TEQs).

Edible marine species (fish, cephalopod molluscs, crustaceans) from the Adriatic Sea were analyzed for content in heavy metals (Hg, Cd and Pb) and polychlorinated biphenyls (PCBs). Health risks to human via dietary intake of seafood were assessed by the target hazard quotients (THQs) and the toxic equivalent factors (TEFs). Mercury maximum concentrations corresponded to fish (0.07-1.56 microg g(-1)w.w.), followed by cephalopod molluscs (0.10-0.55 microg g(-1)w.w.), and crustaceans (0.27-0.33 microg g(-1)w.w.). Cadmium levels in cephalopods (0.18-0.59 microg g(-1)w.w.) were higher than those in fish (0.01-0.05 microg g(-1)w.w.) and crustaceans (0.02-0.04 microg g(-1)w.w.), while for Pb the concentrations were generally low (fish: ND-1.18 microg g(-1)w.w., cephalopods: ND-0.17 microg g(-1)w.w., crustaceans: ND-0.03 microg g(-1)w.w.). For PCBs, concentrations in fish, cephalopods and crustaceans ranged between 141 and 3,406 ng g(-1)l.w., 190 and 542 ng g(-1)l.w., and 202 and 429 ng g(-1)l.w., respectively. Cd and Pb THQ values as well as estimates of PCB TEQ exposure indicated the absence of health risks through consumption of the various seafood. In contrast, mercury TEQs values due to consumption of certain fish species (albacore, rosefish and thornback ray) indicated that human health risk might be of concern.

Persistent organic pollutants (PCBs and DDTs) in small size specimens of bluefin tuna (Thunnus thynnus) from the Mediterranean Sea (Ionian Sea).

The concentration levels and pattern distribution of polychlorinated biphenyls (PCBs) and organochlorine pesticides (DDTs), were investigated in liver of small size specimens of bluefin tuna (Thunnus thynnus) from Mediterranean Sea (Ionian Sea). PCB concentrations (average: 526 ng g(-1) lipid wt) were comparable with DDT levels (average: 435 ng g(-1) lipid wt). The specific analysis of PCB congeners revealed a profile dominated by hexa-, penta- and heptachlorinated congeners, while among DDTs, the compounds with the higher concentration was p,p'-DDE, constituting 91.9% of the total DDT. The total 2,3,7,8-TCDD toxic equivalent (TEQs) of coplanar PCBs, including mono- and non-ortho congeners, was 0.55 pg g(-1) wet weight with. The congeners with highest TEQs values were non-ortho congeners followed by mono-ortho ones.

Total and subcellular distribution of trace elements (Cd, Cu and Zn) in the liver and kidney of green turtles (Chelonia mydas) from the Mediterranean Sea.

This study investigated the subcellular distribution of Cd, Cu and Zn in liver and kidney of green turtles (Chelonia mydas) stranded along the Italian coast of the South Adriatic Sea (Eastern Mediterranean). Cd and Zn mean concentrations did not differ significantly between liver (4.26microgg(-1) and 34.53microgg(-1), respectively) and kidney (5.06microgg(-1) and 26.39microgg(-1), respectively), whereas the levels of Cu were significantly higher in liver (32.75microgg(-1)) than in kidney (8.20microgg(-1)) (p<0.009). Most of Cd, Cu and Zn was present in hepatic and renal cytosol, and their concentrations increased with total levels in both organs, indicating that cytosol has a crucial role in metal accumulation. Cd and Cu in hepatic and renal cytosol were present mostly in metallothionein fractions (MTs), whereas Zn was fractionated into MTs and high-molecular-weight-substances (HMWS). The comparison with the results of other investigations on individuals of the same species collected in different marine areas shows good agreement relatively to essential metals. For Cd our data are comparable with those encountered in specimens from the Mediterranean Sea (Cyprus) confirming the homogeneity of the area comprising the south-eastern basin of the Mediterranean Sea from an ecological point of view.

Residues of polychlorinated biphenyls in edible fish of the Adriatic Sea: assessment of human exposure.

Food consumption is an important route of human exposure to polychlorinated biphenyls (PCBs). To assess the potential health risks associated with these contaminants due to fish consumption, the muscle tissue of edible species of fish was analyzed. Contamination levels among the different species varied from 134 to 1210 ng/g lipid weight. Isomer-specific analysis revealed a profile dominated by hexa- (57.9% to 82.9%) and pentachlorobiphenyls (10.3% to 23.9%), followed by hepta- (9.0% to 19.2%) and tetrachlorobiphenyls (0.7% to 17.5%). Other congeners, including those with less than 4 or more than 9 chlorine atoms, were below the instrumental limit of detection in all samples. Risk evaluation for human health was carried out by comparing the experimental data with the new European Commission legal level and a new approach proposed by the U.S. EPA to protect population-segments who repeatedly consume fish. The dietary intake of PCBs, as WHO-TEQ per kg body weight (b.w.), was below 8 pg TEQ/kg body weight/week, while the new EPA approach suggested that the chronic effects do not represent any type of danger for human health, while the possible rise in the carcinogenic risk connected with consumption of some type of fish is more worrisome.

Mercury in fish: concentration vs. fish size and estimates of mercury intake.

Total mercury concentrations were determined in different fish size classes of commercial importance such as, conger eel (Conger conger), starry ray (Raja asterias), forkbeard (Phycis blennoides), frostfish (Lepidopus caudatus), striped mullet (Mullus barbatus), red gurnard (Aspitrigla cuculus) and yellow gurnard (Trigla lucerna) in order to evaluate variations in consumer exposure to mercury as a function of fish consumption of a spectrum of different sizes. The highest mean levels of total mercury were detected in conger eel (0.80 microg g(-1)) and starry ray (0.75 microg g(-1)). Forkbeard (0.67 microg g(-1)), frostfish (0.59 microg g(-1)) and striped mullet (0.55 microg g(-1)) showed slightly lower levels, while red gurnard (0.33 microg g(-1)) and yellow gurnard (0.22 microg g(-1)) exhibited the lowest concentrations. The results of linear regression analysis showed a significant relationship between mercury concentrations and fish size for all species. Consequently, dietary consumption of larger size specimens leads to an increase in the exposure level for consumers. Understanding by consumers of all factors leading to an increase of exposure to mercury is the first step to enable them to make decisions about eating fish.

Polychlorinated biphenyls and other chlorinated organic contaminants in the tissues of Mediterranean loggerhead turtle Caretta caretta.

Polychlorinated biphenyls including coplanar congeners and DDT compounds were measured in different organs and tissues (liver, kidney, lung and muscle tissue) of loggerhead turtles Caretta caretta from the eastern Mediterranean Sea. The highest levels of these compounds were found in liver (PCBs: 52.32 ng/g; DDTs: 18.27 ng/g), followed by kidney (PCBs: 19.05 ng/g; DDTs: 5.70 ng/g), lung (PCBs: 12.75 ng/g; DDTs: 3.76 ng/g ) and muscle tissue (PCBs: 4.65 ng/g; DDTs: 1.45 ng/g). PCBs revealed a profile dominated by hexa-, penta- and hepta-chlorinated congeners, while among DDTs, the compound in the greatest concentration was p,p'-DDE, (liver: 85.2%, kidney: 93.6%, lung: 86.4%, muscle tissue: 93.2%). The estimated toxic equivalents (pg TEQs/g wet wt) of non- and mono-ortho PCBs were in the range of 1.54-5.86 pg TEQs/g wet wt. Non-ortho coplanar PCB 77 accounted for more than 90% of the total TEQs leaving to mono-ortho only 2.6-6.2%.

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.

Occurrence and accumulation of organochlorine contaminants in swordfish from Mediterranean Sea: a case study.

Polychlorinated biphenyls including coplanar congeners and DDT compounds were measured in the liver of a teleost species: namely, Xiphias gladius. PCB concentrations (median: 1121 ng/g lipid wt) were comparable with DDT levels (median: 1236 ng/g lipid wt). PCBs revealed a profile dominated by hexa-, penta- and heptachlorinated congeners. Among DDTs, the compound in the greatest concentration was p,p'-DDE, representing 70% of the total DDT burden, followed by o,p'-DDT>p,p'-DDT>p,p'-DDD=o,p'-DDD. Mean total 2,3,7,8-TCDD equivalent of five coplanar PCBs was 8.83 pg/g lipid weight. The isomers with higher TEQs values were non-ortho congeners than mono-ortho ones.

Concentrations and composition of organochlorine contaminants in different species of cephalopod molluscs from the Italian waters (Adriatic Sea).

Persistent organochlorines, such as polychlorinated biphenyls including coplanar congeners and DDT compounds were measured in the liver of different species of mollusc cephalopods: broadtail squid (Illex coindetii), horned octopus (Eledone moschata) and spider octopus (Octopus salutii) collected in two different areas of the Adriatic Sea (central and southern Adriatic Sea). The concentration of PCBs exceeded DDT ones in horned octopus (PCBs: 617.0 ngg(-1) lipid weight, DDTs: 188.0 ngg(-1) lipid weight) and in spider octopus (PCBs: 590.0 ngg(-1) lipid weight, DDTs: 293.0 ngg(-1) lipid weight), while in broadtail squid PCB load (748.0 ngg(-1) lipid weight) was similar to those of DDTs (514.5 ngg(-1) lipid weight). No significant difference in PCB levels was observed among species, while DDT concentrations were higher in broadtail squid than in horned octopus and spider octopus. DDTs composition showed slight differences in relation to species, thought p,p'-DDE was the predominant compound in all cephalopod species (broadtail squid: 88.1%, horned octopus 94.1%, spider octopus: 97.5%). PCB isomer profiles were similar among species being hexachlorinated isomers the most abundant, followed by penta- and heptachlorobiphenyls, while tri- and tetrachlorobiphenyls made up a small percentage of the total PCB residues. Regarding coplanar congeners, non-ortho PCBs were below the detection limit in all samples and the 2,3,7,8-TCDD toxic equivalent (TEQ) concentrations were very low. The influence of biological and ecological factors (size/weight and location) on the bioaccumulation of organochlorine compounds has been discussed.