Evaluation of relationships between reproductive metrics, gender and vitellogenin expression in demersal flatfish collected near the municipal wastewater outfall of Orange County, California, USA.

Estrogenic activity in fish has primarily been evaluated using vitellogenin (vtg) expression in male and juvenile animals. Although the response has been widespread in field and laboratory studies, the relevance of the response to higher level adverse effects, particularly in the field, is less than clear. Previous evaluations of vtg within flatfish species collected near the Orange County Sanitation District (OCSD) outfall and stations as far as 7.7 km down current indicated bioavailable estrogens within demersal flatfish populations. In order to evaluate the persistence of estrogenic activity and relationships to reproduction and development, fish were sampled in the winter and summer of 2003 and 2004 at the outfall and a reference location. Vtg, plasma estradiol (E2) concentrations, gonadosomatic indices (GSI), sperm DNA damage, development, and gender ratios were measured in English Sole (Pleuronectes vetulus) and Hornyhead Turbot (Pleuronichthys verticalis). Variable levels of vtg were continually observed in the plasma samples of fish collected at both locations. Vtg expression and plasma E2 levels were significantly correlated in females. A positive relationship was demonstrated between plasma E2 levels and sperm DNA damage. Rather than an expected feminization of populations, a trend toward masculinization was observed particularly at the OCSD outfall, as indicated by gender ratios and significantly higher GSI in males versus females. These results are consistent with previous studies showing vtg expression in male flatfish, but no alteration in overall flatfish abundance at the sampled sites.

In vivo bioassay-guided fractionation of marine sediment extracts from the Southern California Bight, USA, for estrogenic activity.

The exposure and uptake of environmental estrogenic compounds have been reported in previous studies of demersal flatfish species in the central Southern California Bight (SCB), USA. The objective of this study was to evaluate the estrogenic or feminizing activity of marine sediments from the SCB by using in vivo vitellogenin (VTG) assays in male or juvenile fish. In 2003, sediments were collected near wastewater outfalls serving the counties of Los Angeles (LACSD) and Orange (OCSD), and the city of San Diego (SD), California, USA. Cultured male California halibut (CH; Paralichthys californicus) were either directly exposed to sediments for 7 d or treated with two intraperitoneal injections of sediment extract over 7 d. The 17beta-estradiol (E2) equivalent values ranged from 1 to 90 microg/kg with LACSD > SD > OCSD. Measurable concentrations of E2 were observed in all sediment extracts and ranged from 0.16 to 0.45 ng/g. Estrone (El) was only observed in sediments near the LACSD outfall (0.6 ng/g). Alkylphenols and alkylphenol ethoxylates were observed in all sediment samples, but were highest near the OCSD outfall, where concentrations of nonylphenol were 3,200 ng/g. Fractionation studies of the LACSD sediment extract collected in 2004 failed to demonstrate relationships between VTG expression and 62 analytes, including E2, which was observed in the whole extract (2.9 ng/g). Oxybenzone (1.6 ng/g) was identified in bioactive fractions as well as unknown compounds of relatively high polarity. These results indicate that estrogen receptor-based assays may underestimate environmental estrogenic activity and estrogenic compounds other than classic natural and xenoestrogens may contribute to estrogenic activity of sediments from the SCB.