Daphnia magna and ecotoxicogenomics: gene expression profiles of the anti-ecdysteroidal fungicide fenarimol using energy-, molting- and life stage-related cDNA libraries

In the present study, the existing life stage-specific cDNA library was extended with energy- and molting-related genes using Suppression Subtractive Hybridization PCR and a microarray for the aquatic test organism Daphnia magna was created. A gene set of 2455 fragments was produced belonging to different pathways such as carbohydrate and lipid metabolism, O2 transport and heme metabolism, immune response, embryo development, cuticula metabolism and visual perception pathways. Using this custom microarray, gene expression profiles were generated from neonates exposed to three concentrations of the anti-ecdysteroidal fungicide fenarimol (0.5, 0.75, 1 microg/ml) during 48 h and 96 h. In total, 59 non-redundant genes were differentially expressed, of which more genes were down- than up-regulated. The gene expression data indicated a main effect on molting specific pathways. At the highest concentration, a set of proteolytic enzymes - including different serine proteases and carboxypeptidases - were induced whereas different cuticula proteins were down-regulated (48 h). Moreover, effects on embryo development were demonstrated at the gene expression as well as at the organismal level. The embryo development related gene vitellogenin was differentially expressed after 96 h of exposure together with a significant increase in embryo abnormalities in the offspring. This study suggests that this Daphnia magna microarray is of great further value for the elucidation of molecular mechanisms of toxicity and for the future development of specific biomarkers for hazard characterization.     

Screening of thyroid gland histology in organohalogen-contaminated glaucous gulls (Larus hyperboreus) from the Norwegian Arctic

The associations between blood organohalogen contaminant (OHC) concentrations and thyroid gland histology were studied in 10 adult female glaucous gulls (Larus hyperboreus) from the Norwegian Arctic (Bjørnøya) during the incubation period. This histological investigation was undertaken as previous glaucous gull studies from the same area reported negative relationships between circulating OHC concentrations and thyroid hormone levels. Organohalogen concentrations have previously been associated with altered blood plasma thyroid hormone concentrations, as a result of parenchymal thyroid gland alterations and perturbation of the hypothalamic-pituitary-thyroid (HPT)-axis. In this study, PCB (range: 186–1027 ng g^?1ww), DDT (77–203 ng g^?1ww) and chlordane (18–65 ng g^?1ww) concentrations dominated the blood plasma OHC profile in incubating female glaucous gulls. High density of small follicles accompanied by follicular epithelial cell proliferations was seen in thyroid glands in seven of 10 gulls. Focal thyroiditis and nodular hyperplasia were found in two birds. No significant differences in plasma OHC concentrations were noted between gulls exhibiting high density of small follicles and cell proliferations and those birds not showing histological changes. Based on these findings, data suggest that the histological changes in thyroid glands of OHC-contaminated female glaucous gulls may be due to natural variance, although an OHC-induced thyroid stimulating hormone (TSH) perturbation resulting in epithelial cell hyperplasia and increased follicular density cannot be ruled out and remains to be verified. Hence, a large-scale histological study is required, in order to elaborate the potential linkage between changes in thyroid gland histology, OHC exposure and regulation of the HPT-axis in the Arctic-breeding glaucous gull.