No abstract

In response to the initial Endocrine Disruptor Screening and Testing Advisory Committee (EDSTAC) recommendations, research was conducted on the development of a Xenopus laevis based tail resorption assay for evaluating thyroid axis disruption. This research highlighted key limitations associated with relying on tail resorption as a measure of anti/thyroid activity. The most critical limitation being that tail tissues of tadpoles at metamorphic climax are insensitive to perturbation by thyroid axis agonists/antagonists. To improve upon the initial proposal, we have conducted experiments comparing the sensitivity of pre-metamorphic (stage 51) and pro-metamorphic (stage 54) larvae to the model thyroid axis disruptors methimazole (control, 6.25, 12.5, 25, 50, 100 mg/l), 6-propylthiouracil (PTU) (control, 1.25, 2.5, 5, 10, and 20 mg/l), and thyroxine (T4) (0.25, 0.5, 1, 2, 4 microg/l). Exposures were conducted using two different experimental designs. For experimental design 1, tadpoles were exposed to methimazole or PTU starting at either NF stage 51 or NF 54 for 14 days. For experimental design 2, tadpoles were exposed to PTU or T4 starting at NF stage 51 or NF 54 for 14 and 21 days, respectively. Methimazole and PTU, which are thyroid hormone synthesis inhibitors, both caused a concentration dependent delay in larval development. As determined from this endpoint, there were only minor differences in sensitivity observed among the two stages examined. Further, both compounds caused concentration dependent changes in thyroid gland morphology. These changes were characterized as reduced colloid, glandular hypertrophy, and cellular hyperplasia and hypertrophy. Treatment failed to negatively affect growth, even in tadpoles that experienced significant metamorphic inhibition. T4 treatment resulted in a concentration dependent increase in developmental rate, as would be expected. Similar to studies with methimazole, there were no differences in sensitivity among the two developmental stages examined. These results indicate that tadpoles in the early stages of metamorphosis are sensitive to thyroid axis disruption and that development of a short-term, diagnostic amphibian-based thyroid screening assay shows considerable promise.

We investigated the influence of sulfide, measured as acid-volatile sulfide (AVS), on the bioavailability of cadmium and nickel in sediments. Seventeen samples from an estuarine system heavily contaminated with cadmium and nickel were analyzed for AVS and simultaneously extracted metals (SEM) and tested in 10-d exposures with the amphipod HyaleNa azteca and the oligochaete Lumbriculus varregatus. Molar SEM (cadmium + nickel)/AVS ratios in the sediments ranged from less than one to greater than 200, with several in the range of 1 to 10. Samples with SEM/AVS ratios greater than one were consistently toxic to Hyalella azteca, whereas sediments with ratios less than one were not. Lumbriculus variegatus was less sensitive to the test sediments than Hyalella azteca, which was consistent with their relative sensitivity to cadmium and nickel in water-only exposures. SEMIAVS ratios in the sediments also appeared to be important in determining bioaccumulation of metals by Lumbrimlus variegatus. These results support other studies with metal-spiked samples in demonstrating the importance of AVS in determining metal bioavailability in sediments and suggest that AVS normalization is a reasonable means for assessing the hazard of some sediment-associated metals to aquatic ecosystems.

Standard 10 d toxicity tests were conducted with freshwater benthic invertebrates using sediments containing a range of concentrations of polycylic aromatic hydrocarbons (PAHs) The assays were performed both under normal laboratory fluorescent light and ultraviolet (UV) light, which mimicked wavelengths present in sunlight, at about 10% of ambient solar intensity In sediments with elevated PAH concentrations, tests conducted with UV light resulted in significantly greater mortality of Hyalella azteca (amphipods) and Lumbriculus variegatus (oligochaetes) than tests performed under otherwise comparable conditions with fluorescent light There also was increased mortality of these two species, relative to controls, when surviving organisms from the 10‐d exposures to the PAH contaminated sediments were placed in clean water under UV light for 2 h These results suggest that the organisms accumulated PAHs from the test sediments, which were subsequently photoactivated by UV light to excited states more toxic than the ground state molecules The phenomenon of photoactivation has been examined for pelagic species exposed to PAHs, but not for benthic organisms exposed to sediment‐associated PAHs Our results suggest that failure to consider photoactivation of PAHs by sunlight could result in sediment toxicity test methods or criteria that are underprotective of benthic organisms

The perchlorate anion inhibits thyroid hormone (TH) synthesis via inhibition of the sodium-iodide symporter. It is, therefore, a good model chemical to aid in the development of a bioassay to screen chemicals for affects on thyroid function. Xenopus laevis larvae were exposed to sodium perchlorate during metamorphosis, a period of TH-dependent development, in two experiments. In the first experiment, stage 51 and 54 larvae were exposed for 14 d to 16, 63, 250, 1,000, and 4,000 microg perchlorate/ L. In the second experiment, stage 51 larvae were exposed throughout metamorphosis to 8, 16, 32, 63, and 125 microg perchlorate/L. Metamorphic development and thyroid histology were the primary endpoints examined. Metamorphosis was retarded significantly in the first study at concentrations of 250 microg/L and higher, but histological effects were observed at 16 microg/L. In the second study, metamorphosis was delayed by 125 microg/L and thyroid size was increased significantly at 63 microg/L. These studies demonstrate that inhibition of metamorphosis readily can be detected using an abbreviated protocol. However, thyroid gland effects occur at concentrations below those required to elicit developmental delay, demonstrating the sensitivity of this endpoint and suggesting that thyroidal compensation is sufficient to promote normal development until perchlorate reaches critical concentrations.