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The aim of this study was to assess the potential genotoxic effects of produced water (PW) from an Italian on-shore oil plant. Produced water is a complex mixture containing residual hydrocarbons, trace elements, naturally occurring radioactive material and potentially toxic treatment chemicals such as biocides, dispersants, detergents and scale inhibitors used in oil production. The test organism, mosquito fish (Gambusia affinis), was divided into male and female groups and exposed for 8 days in the laboratory to 50% concentrations of different produced waters: PW before treatment and after settling treatment. The fish were also exposed to lower concentrations (10%) of the same PW for 30 days. DNA damage was evaluated in erythrocytes by single cell gel electrophoresis (Comet assay) and micronucleus test, while an oxidative stress biomarker, was assessed. Polycyclic aromatic hydrocarbons (PAH) metabolites in bile were also evaluated. A higher sensitivity in biomarker responses was found in females in comparison to males. An increase in DNA strand breaks was observed in both genders after 30 days exposure and a statistically significant increase of micronucleated cells was found in females after 8 days exposure. A positive correlation between presence of micronucleated cells and PAH metabolites in bile was also observed.

The aim of the present study was to propose the immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as a new “in vitro” tool to explore the susceptibility of these marine mammals to different xenobiotic compounds. The cell lines were cultured from integument biopsies of free-ranging and stranded cetaceans (dead within 12 h). Using the indirect immunofluorescence assay, we detected endogenous proteins induced by different contaminants. Here we present the method used for qualitative and quantitative evaluation of cytochromes P450 (CYP1A1 and CYP2B) induced by some POPs (DDTs and PCBs) and emerging contaminants (PBDEs) in fibroblast cell cultures of striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Immunofluorescence was quantified with a specially designed Olympus macro, DetectIntZ. A major result was the possibility of using this “in vitro” assay to quantify induction of endogenous proteins

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SUMMARY: This paper reports a histological, histochemical and immunohistochemical characterisation of growing oocytes of the swordfish Xiphias gladius. The presence and distribution of carbohydrates, proteins, lipids, calcium, iron, vitellogenin/ Vg, zona radiata protein/Zrp, metallothionein/Mt, and thyroid hormones/T 3 -T 4 were studied during oogenesis (cortical alveoli, globules, yolk-granules, cytoplasm, follicular and radiata envelopes). During the initial vitellogenic phase, the oocytes showed cortical alveoli and oil globules containing neutral lipids exclusively. During this phase, small yolk granules appeared around the peripheral cytoplasm, and they increased through exogenous vitellogenesis. Yolk granules were composed of glycoproteins, calcium, iron, and proteins rich in lysine, arginine, tyrosine, tryptophan, cysteine and cystine. Vg and Mt were immunohistochemically detected in yolk. The follicular envelope contained proteins rich in amino acids. Moreover, calcium and thyroid hormones (triiodotyronine and thyroxine/T 3 , T 4 ) were detected in this cell envelope. Cortical alveoli, which contained carboxylated and neutral glycoconjugates, were especially rich in N-acetyl-D-galactosamine, N-acetyl-Dglucosamine, galactose and sialic acid. Finally, the zona radiata was mainly proteinaceous in nature and was composed of calcium and neutral glycoproteins. The egg envelope or chorion and the liver showed specific immunoreactivities by using anti-salmon Zrp as the primary antiserum.Keywords: histochemistry, carbohydrates, lipids, glycoconjugates, vitellogenin, zona radiata protein, methallothionein, thyroid homones, cations, oocytes, Xiphias gladius. RESUMEN: Caracterización histoquímica de ovocitos del pez espada, Xiphias gladius. -En este trabajo se realiza una caracterización histológica, histoquímica e inmunohistoquímica de los ovocitos en crecimiento del pez espada, Xiphias gladius. Se estudia la presencia y distribución de carbohidratos, proteínas, lípidos, calcio, hierro, vitelogenina/Vg, proteína radiada/Zrp, metalotioneina/Mt y hormonas tiroideas/ T 3 -T 4 durante la ovogénesis (alveolos corticales, glóbulos, gránulos-vitelo, citoplasma, capas folicular y radiada). Durante la fase inicial de la vitelogénesis, los ovocitos presentan alveolos corticales y una gota lipídica conteniendo exclusivamente lípidos neutros. Durante esta fase en la periferia del citoplasma aparecen pequeños granulos de vitelo, incrementando progresivamente durante la vitelogénesis exógena. Los gránulos de vitelo están compuestos por glicoproteínas, calcio, hierro, proteínas ricas en lisina, arginina, tirosina, triptófano, cisteina y cistina, y contienen Vg y Mt. La capa folicular de los ovocitos del pez espada está compuesta por glicoproteínas y proteínas ricas en diferentes aminoácidos. Esta envuelta contiene calcio y hormonas tiroideas/ T 3 y T 4 . Los alveolos corticales están constituidos por glicoconjugados carboxilados y neutros, y son especialmente ricos en N-acetil-D-galactosamina, N-acetil-D-glucosamina, galactosa y...