Microarray analysis of di-n-butyl phthalate and 17α ethinyl-oestradiol responses in three-spined stickleback testes reveals novel candidate genes for endocrine disruption.

Phthalate esters are plasticizers frequently found in wastewater effluents. Previous studies on phthalates have reported anti-androgenic activity in mammals, causing concerns of their potential effects on the reproduction of aquatic organisms. Another group of environmental endocrine disrupters, steroidal estrogens, are known to inhibit steroid biosynthesis in the gonads, but the effects related to spermatogenesis are not well understood in fish. In this study, three-spined sticklebacks were exposed to di-n-butyl phthalate (DBP) and 17α ethinyl-oestradiol (EE2) at nominal concentrations 35µg/L and 40ng/L, respectively, for four days. The aim of the study was to obtain insight into the acute transcriptional responses putatively associated with endocrine disruption. RNA samples from eight individual male fish per treatment (including controls) were used in microarray analysis, covering the expression of approximately 21,000 genes. In the EE2 treatment the results show transcriptional downregulation of genes associated with steroid biosynthesis pathway and up-regulation of genes involved in pathways related to epidermal growth factor signaling and xenobiotic metabolism. The transcriptional response to DBP was in general weaker than to EE2, but based on enrichment analysis, we suggest adverse effects on retinoid metabolism, creatine kinase activity and cell adhesion. Among the genes showing highest fold changes after DBP treatment compared to control was the teleost fish -specific cytochrome P450 17A2. Overall, this study promotes our understanding on molecular responses to anti-androgens and estrogens in fish testes.

Direct effects of heavy metal pollution on the immune function of a geometrid moth, Epirrita autumnata.

Previously, we found that Epirrita autumnata larvae, which were fed with leaves that came from a metal polluted area, had an enhanced immune function. To investigate further the cause of the enhanced immunity we examined experimentally the direct influence of the heavy metals, copper and nickel, on the immune function of the geometrid moth we artificially added the metals to the surface of birch leaves to examine their direct effects. As a measurement of the strength of immune function, we used encapsulation rate against a nylon monofilament. A moderate amount of Ni and Cu in the diet of moth larvae increased their encapsulation rate, but a large amount of Cu caused the immune function to decrease. The results indicate that Ni and Cu have direct effects on immunity. Moderate amounts of those metals increase immune function, but large amounts lead to inhibition of immune function.