Pipping Success, Isomer-Specific Accumulation, and Hepatic mRNA Expression in Chicken Embryos Exposed to HBCD

Hexabromocyclododecane (HBCD) is an additive flame retardant used primarily in polystyrene foams. HBCD is a persistent contaminant that has been detected in abiotic and biotic matrices, including wild avian species. The toxicological implications of exposure are not well characterized. We recently identified molecular end points responsive to HBCD exposure in chicken embryonic hepatocytes (CEHs) including genes involved in xenobiotic metabolism, thyroid hormone transport, and lipid metabolism. In the current study, a technical mixture of HBCD (HBCD-TM), comprising 12% alpha-, 11% beta-, and 77% gamma-stereoisomers, was injected into the air cell of chicken eggs prior to incubation. Embryonic viability to pipping, isomer-specific HBCD accumulation, and hepatic mRNA expression of the genes identified in the in vitro study were determined. Concentrations of 100 and 10,000 ng/g decreased pipping success while 50, 300, and 1000 ng/g had no effect. In contrast to HBCD-TM, the isomeric composition in liver tissue was significantly different for alpha- (31%) and gamma-HBCD (61%) demonstrating that isomer-specific processes were occurring in the egg and/or developing embryo. Exposure to 1000 ng/g HBCD-TM significantly upregulated cytochrome P450 (CYP) 2H1, CYP3A37, uridine 5'-diphospho-glucuronosyltransferase, and deiodinase 2, while liver fatty acid-binding protein and insulin-growth factor 1 expression were significantly decreased at 100 and 10,000 ng/g, respectively. The alterations in hepatic mRNA levels were in concordance with those observed in CEH highlighting the utility of both approaches for identifying molecular mechanisms of action. Research on the effects of HBCD in wild avian species is warranted.