Arsenic species and their accumulation features in green turtles (Chelonia mydas)

Total arsenic (As) and its compounds were determined in liver, kidney, muscle, and stomach contents of green turtles (Chelonia mydas). Total As concentrations in the muscle were higher than those in the kidney and liver. Arsenobetaine (AB) was the predominant compound in all the three tissues and its levels were positively correlated with total As concentrations. This indicates that AB greatly contributes to As accumulation in green turtles. Higher concentrations of remaining As in the sample after extraction were detected in the liver, implying that lipid-soluble or protein bound As compounds accumulate in the liver of green turtles. Total As levels in tissues showed significant negative correlations with standard carapace length. The size-dependence of As accumulation in green turtles may be related to their feeding habit, shifting from carnivore to herbivore at different growth stages. Concentrations of AB and dimethylarsinic acid (DMA) were low in the stomach contents but high in the tissues, implying bioaccumulation of these arsenicals in green turtles.     

Toxicogenomic analysis of immune system-related genes in Japanese flounder (Paralichthys olivaceus) exposed to heavy oil

Heavy oil contamination is one of the most important environmental issues. Toxicities of polycyclic aromatic hydrocarbons (PAHs), including immune toxicities, are well characterized, however, the immune toxic effects of heavy oil, as a complex mixture of PAHs, have not been investigated. In the present study, we selected Japanese flounder (Paralichthys olivaceus) as a model organism, and observed alteration of immune function by the exposure to heavy oil. To analyze the expression profiles of immune system-related genes, we selected 309 cDNAs from our flounder EST library, and spotted them on a glass slide. Using this cDNA array, alteration of gene expression profiles was analyzed in the kidneys of flounders exposed to heavy oil. Six Japanese flounders (mean body weight: 197 g) were acclimated to laboratory conditions at 19–20 °C. Three fish were exposed to heavy oil C (bunker C) at a concentration of 3.8 g/L for 3 days, and the others were kept in seawater without heavy oil and used as the control. After the exposure period, the fish were transferred into control seawater and maintained for 4 days, and then they were dissected and their kidneys were removed. Total RNA was extracted from the kidney samples to use in gene expression analyses. The microarray detected alteration of immune system-related genes in the kidneys of heavy oil-exposed flounders, including down-regulation of immunoglobulin light chain, CD45, major histocompatibility complex class II antigens and macrophage colony-stimulating factor precursor, and up-regulation of interleukin-8 and lysozyme. These results suggest that pathogen resistance may be weakened in heavy oil-exposed fish, causing a subsequent bacterial infection, and then proinflammatory genes may be induced as a defensive response against the infection. Additionally, we found candidate genes for use as biomarkers of heavy oil exposure, such as N-myc downstream regulated gene 1 and heat shock cognate 71 kDa proteins.     

Catalytic function of avian cytochrome P450 1A4 and 1A5 (CYP1A4 and CYP1A5) enzymes heterologously expressed using in vitro yeast system

The present study clarifies the enzymatic properties of two avian cytochrome P4501A (CYP1A) paralogs, CYP1A4 and 1A5, using a yeast-based vector system. Recombinant CYP1A4 and 1A5 proteins from common cormorant (Phalacrocorax carbo) were expressed in yeast cells, and showed typical reduced CO-difference spectra with a peak at 446 nm. Kinetic analysis of O-dealkylase of methoxy-, ethoxy-, pentoxy- and benzyloxyresorufin catalyzed by the CYP1A enzymes revealed that Vmax value for ethoxyresorufin-O-deethylase (EROD) activity was much higher than that for the other three O-dealkylase activities for both isozymes. Interestingly, remarkable substrate specificity of the CYP1As was observed for O-dealkylation of benzyloxyresorufin and methoxyresorufin; CYP1A4 was highly specific for catalyzing benzyloxyresorufin-O-debenzylase activity, whereas CYP1A5 was more efficient in catalyzing methoxyresorufin-O-demethylase activity. The present study also measured CYP1A-dependent EROD activity in the presence of 2,3,7,8-tetrachlorodibenzofuran (TCDF) to evaluate the ability of this dioxin-like congener to inhibit the EROD activity. One hundred nanomolar TCDF noncompetitively inhibited CYP1A5-dependent EROD activity, although no inhibitory effect was detected for CYP1A4-dependent EROD activity. These results indicate that the avian CYP1A paralogs have different affinities for substrate and inhibitor, thus suggesting their distinct physiological and toxicological roles.     

Interelement relationships and age-related variation of trace element concentrations in liver of striped dolphins (Stenella coeruleoalba) from Japanese coastal waters

Concentrations of 19 trace elements (V, Cr, Mn, Fe, Co, Cu, Zn, Se, Rb, Sr, Mo, Ag, Cd, Sb, Cs, Ba, Tl, Hg, and Pb) were determined in the liver of the striped dolphins (Stenella coeruleoalba) collected around Japan during 1977-1982 to examine the sex difference, age dependence, and interrelationships among trace elements. Tissue distribution of trace elements was also investigated in one adult and one fetus specimens. Generally, concentrations of Se, Sr, Ag, Cd, Cs, Ba, Hg, and Pb were higher in the tissues of adult than those of fetus, whereas the opposite trend was observed for Cr and Tl. There were no significant sex differences in the trace element levels in the liver. Significant positive correlations between age (0-26.5 years) and hepatic concentrations were found for Ag, Se, Hg, V, Fe, Pb, and Sr, suggesting their age-dependent accumulation in the liver. In contrast, hepatic concentrations of Mn and Zn decreased with age. Significant positive relationships were observed between Se, and Hg, Ag, V, Fe, and Sr in the liver.     

Interelement relationships and age-related variation of trace element concentrations in liver of striped dolphins (Stenella coeruleoalba) from Japanese coastal waters

Concentrations of 19 trace elements (V, Cr, Mn, Fe, Co, Cu, Zn, Se, Rb, Sr, Mo, Ag, Cd, Sb, Cs, Ba, Tl, Hg, and Pb) were determined in the liver of the striped dolphins (Stenella coeruleoalba) collected around Japan during 1977–1982 to examine the sex difference, age dependence, and interrelationships among trace elements. Tissue distribution of trace elements was also investigated in one adult and one fetus specimens. Generally, concentrations of Se, Sr, Ag, Cd, Cs, Ba, Hg, and Pb were higher in the tissues of adult than those of fetus, whereas the opposite trend was observed for Cr and Tl. There were no significant sex differences in the trace element levels in the liver. Significant positive correlations between age (0–26.5 years) and hepatic concentrations were found for Ag, Se, Hg, V, Fe, Pb, and Sr, suggesting their age-dependent accumulation in the liver. In contrast, hepatic concentrations of Mn and Zn decreased with age. Significant positive relationships were observed between Se, and Hg, Ag, V, Fe, and Sr in the liver.