Is normalized hindlimb length measurement in assessment of thyroid disruption in the amphibian metamorphosis assay relevant?

The amphibian metamorphosis assay represents an OECD Level 3 and EDSP Tier 1 ecotoxicity test assessing thyroid activity of chemicals in African clawed frog (Xenopus laevis). To evaluate the effectiveness of snout-vent length (SVL) normalization of hindlimb length (HLL), correlation between the HLL and SVL or body weight was evaluated in the control groups of 10 individual studies from three laboratories. Two studies required separate analysis of the Nieuwkoop-Faber (NF) stage ≤60 and >60 animals creating a total of 12 data sets. On study day 7, significant positive correlation between HLL and SVL or body weight was observed in eight and seven of the 10 data sets, respectively (r = 0.608-0.843 and 0.583-0.876). On study day 21, significant positive correlation between HLL and SVL or body weight was found in three and four of the 12 data sets, respectively (r = 0.452, 0.480 and 0.553 and r = 0.621, 0.546, 0.564 and 0.378). Significant positive correlation between HLL and SVL was found in three of five studies, including ≤NF stage 60 data (r = 0.564, 0.546 and 0.621). In one of eight studies, including >NF stage 60 data, the positive correlation between HLL and body weight was determined (r = 0.378). Negative or no correlation between HLL and SVL or body weight was found in the other late stage data sets. Therefore, use of SVL-normalized HLL to assess thyroid-mediated effects in X. laevis tadpoles is not warranted. HL stage relative to body stage should be considered.

A weight-of-evidence approach for the bioaccumulation assessment of triclosan in aquatic species.

The bioaccumulation assessment of chemicals is challenging because of various metrics and criteria, multiple lines of evidence and underlying uncertainty in the data. Measured in vivo laboratory and field bioaccumulation data are generally considered preferable; however, quantitative structure-activity relationships (QSARs), mass balance models and in vitro data can also be considered. This case study critically evaluates in vivo, in vitro and in silico data and provides new data for the bioaccumulation assessment of triclosan (TCS). The review focusses on measured fish bioconcentration factors (BCFs) because this is the most commonly used regulatory metric. Reported measured fish BCFs range from about 20 to 8700L/kg-ww spanning a range of possible bioaccumulation assessment outcomes, i.e. from "not bioaccumulative" to "very bioaccumulative". Estimated biotransformation rate constants for fish obtained from in vivo, in vitro and in silico methods show general consensus fostering confidence in the selection of plausible values to confront uncertainty in the measured fish BCF tests. Other measurements (lines of evidence) from various species are also collected and reviewed. The estimated biotransformation rate constants and selected chemical property data are used to parameterize bioaccumulation models for aquatic species. Collectively the available lines of evidence are presented using a weight of evidence approach for assessing the bioaccumulation of TCS in aquatic species. Acceptable quality measured data and model predictions for TCS BCFs and bioaccumulation factors are lower than 2000L/kg. Biomagnification factors are <1 (kg/kg). The general consistency in the acceptable quality data is largely explained by the relatively efficient rates of TCS biotransformation in a range of species including measurements of significant in vitro activity of phase II conjugation reactions. The review demonstrates the value of combining models and measurements and, when necessary, applying multiple lines of evidence for chemical assessment.

Effect of triclosan on anuran development and growth in a larval amphibian growth and development assay.

A larval amphibian growth and development assay was performed to evaluate the potential effects of environmentally-relevant concentrations of triclosan (TCS) on amphibian development and growth. Xenopus laevis were exposed to TCS 0.0 (control), 6.3, 12.5 and 25.0 µg l-1 (estimated maximum tolerable concentration) until 10 weeks post-metamorphosis. At median metamorphosis time (Nieuwkoop and Faber stage 62), five larvae per replicate were collected for snout-vent length, hind limb length and body weight measurements, and histopathological examination of thyroid glands. Endpoints evaluated at test termination were based on draft guidance (USEPA, ) and included: survival; snout-vent length; body weight; gender; nuptial pad development (males); and liver, kidney, gonad and gonadal ducts histopathology. Exposure to TCS did not decrease survival, induce general signs of toxicity, affect median metamorphosis time or alter sex ratios. Exposure to TCS 12.5 and 25 µg l-1 increased growth during the metamorphic stages relative to the control, but did not influence growth during the post-metamorphic phase. Overall, several statistically significant findings were found in larvae exposed to TCS, such as a decrease in the prevalence of stage 3 Müllerian ducts in the anterior trunk sections of TCS 25.0 µg l-1 dose group females as compared to controls; most were not considered toxicologically relevant. Copyright © 2017 John Wiley & Sons, Ltd.

Gene toxicity studies on titanium dioxide and zinc oxide nanomaterials used for UV-protection in cosmetic formulations.

Titanium dioxide and zinc oxide nanomaterials, used as UV protecting agents in sunscreens, were investigated for their potential genotoxicity in in vitro and in vivo test systems. Since standard OECD test methods are designed for soluble materials and genotoxicity testing for nanomaterials is still under revision, a battery of standard tests was used, covering different endpoints. Additionally, a procedure to disperse the nanomaterials in the test media and careful characterization of the dispersed test item was added to the testing methods. No genotoxicity was observed in vitro (Ames' Salmonella gene mutation test and V79 micronucleus chromosome mutation test) or in vivo (mouse bone marrow micronucleus test and Comet DNA damage assay in lung cells from rats exposed by inhalation). These results add to the still limited data base on genotoxicity test results with nanomaterials and provide congruent results of a battery of standard OECD test methods applied to nanomaterials.