Bioconcentration, biotransformation and elimination of pyrene in the arctic crustacean Gammarus setosus (Amphipoda) at two temperatures.

The influence of temperature on the bioaccumulation, toxicokinetics, biotransformation and depuration of pyrene was studied in the arctic marine amphipod Gammarus setosus. A two-compartment model was used to fit experimental values of total body burden, total metabolites and parent pyrene concentrations and to calculate toxicokinetic variables derived for two experimental treatments (2 and 8 °C). No statistically significant differences were observed with temperature for these toxicokinetic variables or bioconcentration factors. Contrarily, the Q10 values suggested that the toxicokinetic variables ke and km were temperature-dependent. This may be explained by the high standard deviation of the Q10 values. Q10 is the variation in the rate of a metabolic reaction with a 10 °C increase in temperature. Depuration rate constants were calculated from linear best fit equations applied to measured pyrene concentrations over time during the depuration phase of the experiment. During depuration, the parent pyrene was eliminated in two stages with faster elimination observed at 8 °C compared to 2 °C. This finding was also indicated by the Q10. No changes in total body burdens of metabolite concentrations were observed during the monitoring of depuration over a period of 96 h. The biotransformation pathway of pyrene in G. setosus was also investigated in this study with two main phase II biotransformation products discovered by liquid chromatography. These products are conditionally identified as the sulphate and glucose conjugates of 1-hydroxy-pyrene. Overall, the study contributes new knowledge to the understanding of the fate of PAHs in arctic biota. In particular, the study provides valuable insight into the bioaccumulation and biotransformation of an important PAH and its metabolites in a species that serves as both a predator and prey in the arctic ecosystem.

Trophic transfer of pyrene metabolites and nonextractable fraction from Oligochaete (Lumbriculus variegatus) to juvenile brown trout (Salmo trutta).

The trophic transfer of pyrene metabolites was evaluated by a 2-month exposure of the freshwater annelid Lumbriculus variegatus (Oligochaeta) to pyrene, followed by feeding to juvenile brown trout (Salmo trutta). The results obtained by scintillation counting (SC) proved that the pyrene metabolites produced by L. variegatus were transferred to juvenile S. trutta through diet. More detailed analyses by LC-FLD (liquid chromatography with fluorescence detection) showed that an unknown pyrene metabolite originating from L. variegatus was present in fish liver. This metabolite, although yet not properly identified, may be the glucose conjugate of 1-hydroxy-pyrene. This metabolite was not present in chromatograms of fish that were fed pyrene-spiked food pellets. In addition, the strongly bound tissue residue of L. variegatus, which was nonextractable neither by organic solvents nor by the proteolytic enzyme Proteinase K, was most likely not available for the fish through diet. Altogether, the present study shows that the metabolites of pyrene produced at low levels of the food chain may be potentially available for upper levels through diet, raising a concern about their potential toxicity to predators and supporting their inclusion in the risk assessment of PAHs.

Humic substances modify accumulation but not biotransformation of pyrene in salmon yolk-sac fry.

Humic substances may influence the bioavailability of polycyclic aromatic hydrocarbons (PAHs) in aquatic environment. Relatively little is known how humic substances affect the biotransformation of PAHs in aquatic animals. Here we report how two different types of humic substances affected the accumulation of pyrene, a four-ringed PAH, to yolk-sac fry of landlocked salmon (Salmo salar m. sebago). The accumulation of pyrene to yolk-sac fry tissues was inversely related to humic substance concentration in a short term (72h) exposure. However, the biotransformation of pyrene was not affected by humic substances. Pyrene or humic substances did not induce CYP1A activity in yolk-sac fry tissues contrasting to beta-naphthoflavone, which was used as a positive control. Yolk-sac fry were capable to biotransform pyrene to phase I (1-hydroxypyrene) and phase II (pyrene-1-sulphate) products. Interestingly, glucuronide conjugate (i.e. pyrene-1-glucuronide) was not present in yolk-sac fry tissues. The concentration of parent pyrene and 1-hydroxypyrene remained the same throughout the experiment but the concentration of pyrene-1-sulphate more than doubled from 24 to 72h. This finding suggests that salmon yolk-sac fry are not capable to excrete phase II biotransformation products or the excretion is very slow. Further, this could indicate that early life stage toxicity of many CYP1A inducing compounds is related to accumulation of phase II conjugates in fry tissues.

Toxicity of a phytosterol mixture to grayling (Thymallus thymallus) during early developmental stages.

The study concerns the toxicity of a phytosterol mixture, ultrasitosterol, consisting mainly of beta-sitosterol 75.7% and beta-sitostanol 13.0%, to grayling (Thymallus thymallus) embryos. Eyed eggs were exposed to three concentrations (1 microg/l, 10 microg/l, and 50 microg/l) of ultrasitosterol for 4 weeks. Embryos and later on hatched fry were taken for triiodothyronine (T3), thyroxine (T4), and histopathological analyses after 7, 14, 21, and 28 days of exposure. Most of the eggs (>95%) hatched during the first week of exposure, and ultrasitosterol treatment shortened hatching time significantly (p < or = 0.0001) at all exposure concentrations in comparison to the control. Ultrasitosterol did not have any significant effect on T3 or T4 levels in the embryo extracts. However, an interesting observation was that T3 levels increased in all treatments and in the control near the time of hatching. In conclusion, ultrasitosterol showed potential to affect the development of grayling embryos and fry, but further long-term exposure experiments are needed to verify these changes in more detail.

Toxicokinetics of waterborne bisphenol A in landlocked salmon (Salmo salar m. sebago) eggs at various temperatures.

Eye-pigmented eggs of landlocked salmon were exposed to waterborne [14C]-labeled bisphenol A at four temperatures (2, 6, 8, and 12 degrees C). Both in accumulation and depuration experiments, the eggs were exposed to a bisphenol A concentration of 1 microg/L for 196 h. In the depuration experiment, the exposed eggs were placed into clean water for 96 h. At each sampling time, the eggs were dissected into three parts (eggshell, embryo, and yolk sac), and all of these parts were weighed and analyzed separately in a liquid scintillation counter. The results show that waterborne bisphenol A accumulates in developing salmon eggs. Accumulation of bisphenol A showed a clear trend both in whole eggs and in dissected parts: the higher the temperature, the higher the uptake rate constant and the concentration of bisphenol A. Steady state was reached only in the embryos at the lowest temperature; in other cases, accumulation was linear. Depuration was significant only in the embryos at 2 and 6 degrees C. As the accumulation and the depuration results show, the elimination of bisphenol A was evidently more significant at lower temperatures.