Brain asymmetry as a potential biomarker for developmental TCDD intoxication: a dose-response study.

Previous studies have indicated that in ovo exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds is correlated with the development of grossly asymmetric brains. This asymmetry is manifested as a difference between the two halves of the forebrain and the tecta. Previously, only wildlife species (heron, cormorant, and eagle) had been shown to manifest this response. In the wildlife studies, the frequency and degree of left-right interhemispheric differences had been correlated with the levels of polychlorinated dibenzo-p-dioxin toxic equivalency factors (TEFs) in eggs from the same nest (heron, cormorant). We studied the effect of in ovo exposure to TCDD on the brain throughout development in a sensitive laboratory model (chicken). Embryos from chicken eggs (Gallus gallus) injected with one of several doses of TCDD or vehicle control were sacrificed after 9, 11, 13, 15, 17, or 20 days of incubation, or incubated to hatch and then sacrificed either within 24 hr or at 3 weeks post-hatch. Measurements of both chicken embryo and hatchling brains indicated that 1) TCDD alone induced the brain asymmetry in developing chickens; 2) this brain asymmetry was similar to that observed in animals exposed in the wild to a mixture of TCDD-related contaminants; 3) there was a dose-related increase in both the frequency and severity of brain asymmetry observed at all ages measured; and 4) the asymmetry was measurable in embryonic brains at an age when the braincase was a thin, flexible layer (embryonic day 9), implying that the effect of TCDD was directly on the developing brain and not indirectly via an effect on the braincase.

Morphometric abnormalities in brains of great blue heron hatchlings exposed in the wild to PCDDs.

Great blue heron hatchlings from colonies in the Strait of Georgia, British Columbia, Canada are being monitored for environmental contaminant exposure and effects by the Canadian Wildlife Service. The contaminants of concern are polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), primarily derived from kraft pulp mill effluent. The levels of PCDDs and PCDFs in eggs from the most contaminated colonies peaked in 1988 and 1989 and dropped dramatically through 1990 to 1992. Brains of heron hatchlings (taken as eggs from the wild and hatched in the laboratory) were analyzed for gross morphological abnormalities. Brains from highly contaminated colonies (Crofton, British Columbia and University of British Columbia Endowment Lands) in 1988 exhibited a high frequency of intercerebral asymmetry. The frequency of this abnormality decreased in subsequent years as the levels of TCDD and TCDD-TEQs (toxic equivalence factors) decreased. The asymmetry was significantly correlated with the level of TCDD and TCDD-TEQs in eggs taken from the same nest. Yolk-free body weight negatively correlated and the brain somatic index positively correlated with the TCDD level in such pair-matched eggs. These results indicate that gross brain morphology, and specifically intercerebral asymmetry, may be useful as a biomarker for the developmental neurotoxic effects of PCDDs and related chemicals.

Catecholamine effects on dissociated tiger salamander Muller (glial) cells.

Dissociated Muller (glial) cells from the neotenous tiger salamander retina respond electrogenically and rheogenically to three putative catecholamine (CA) neurotransmitters, epinephrine, norepinephrine and dopamine. All 3 CAs stimulate a net inward current and an increase in input resistance (Rn). The Muller cell response to the CAs is concentration dependent. At high concentrations ascorbate, an antioxidant used to protect the CAs from oxidation, stimulated a net outward current and a decrease in Rn. When the effects of ascorbate were considered, the CA response at 1 mM was no larger than the response at 100 microM, indicating that 100 microM CA maximally stimulated the Muller cell response.

Risk assessment of mercury exposure through fish consumption by the riverside people in the Madeira Basin, Amazon, 1991.

Aquatic food chain mercury pollution is one of the consequences of the gold rush in the Amazon, which started in the late 1970s. This paper addresses the risks of methylmercury (MeHg) toxicity by a riverside population of heavy fish eaters along the Madeira river, in the Amazon, based on their hair mercury (Hg) concentration. Given the vulnerability of the developing nervous system, NOEL/LOEL values were used based on prenatal (LOELp = 0.7 microgram/ kg bw), and adult and childhood (LOELa = 3 micrograms/kg bw) Hg exposures. Based on hair Hg concentrations, we observed that approximately 95% of infants were at risk of absorbing Hg through the previous placental exposure, and/or by ingesting Hg from mother's milk, and/or fish consumption, at a level as great as the LOELp. The hazard quotient derived from the LOELp for neurobehavioral effects was 64 based on an estimated mean Hg daily intake of 4.5 micrograms/kg bw. Approximately 45% of the mothers of the infants and other women of child bearing age were at risk of ingesting Hg at a level equivalent to the LOELp. This also translates into a derived hazard quotient for neurobehavioral effects of 17 for all potential mothers in the population. The non-infant population at the highest risk was fish-eating children under 5 years old. This sub-population had a mean estimated Hg daily intake of 6.4 micrograms/kg bw. This resulted in a probability that almost 60% of this sub-population ingested Hg at a level equivalent to the LOELa or higher. For this sub-population, there was a hazard quotient of 21. These data strongly indicate that the young children of this riverside fish-eating population may be ingesting Hg doses that have been correlated with neurological damage from Hg poisoning.

Suppression of endogenous antioxidant enzymes by 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced oxidative stress in chicken liver during development.

Domestic chickens (Gallus gallus) are an excellent model in which to evaluate developmental toxicity and oxidative stress because of their high sensitivity to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The goal of this study was to measure the effects of environmentally relevant doses of TCDD on endogenous hepatic antioxidant enzyme activity in hatchling chickens. The vehicle (sunflower oil) or 2, 20, or 200 pg/g TCDD was injected into chicken eggs before incubation. On hatching, livers were harvested and quickly frozen. The changes in activity of antioxidant enzymes, including glutathione peroxidase (GPx), glutathione reductase (GRx), copper zinc superoxide dismutase (SOD), and catalase (CAT) were determined as indicators of oxidative stress. TCDD exposure was associated with a significant suppression of the activities of the protective endogenous enzymes GPx, GRx, and SOD in the liver, even at the lowest dose. CAT activity was also suppressed, but not significantly. The measured decreases were 37% to 63% for GPx, 50% to 58% for GRx, 30% to 40% for SOD, and 16% to 24% for CAT. Noncomplex dose-response relationships were evident in GPx and GRx, whereas SOD and CAT curves were U-shaped. These results demonstrate that a decreased ability to scavenge reactive oxygen species may result from developmental TCDD exposure at very low doses, contributing to oxidative stress and thus to the embryotoxicity of TCDD.

Linear regression models of methyl mercury exposure during prenatal and early postnatal life among riverside people along the upper Madeira river, Amazon.

This research is focused on prenatal and early postnatal mercury (Hg) exposure among the riverside people along the Upper Madeira river in the Amazon. Linear regression models were developed to predict the hair Hg concentration in infants. The independent variables included in the model of Group 1 (87 pairs of mothers and their infants) were the average maternal hair Hg concentration and maternal age. Group 2 (31 pairs) included maternal segmental hair Hg concentrations. For the segmental hair Hg analysis over time, it was assumed that hair grows at a rate of 11 cm per month. Thus, information on the timing of the dates of pregnancy and breast feeding from the birth history was used to cut the hair strands into segments, making them correspond to the mother's reproductive stage of life (31 pairs of mothers and their infants). Breast milk Hg concentration results were included with segmental and average maternal hair Hg concentration values (22 and 44 pairs of mothers and their infants, respectively). The models including the breast milk Hg concentration indicated that 61 and 55% of the variability of the infant hair Hg concentrations were due to the independent variables: segmental maternal hair Hg with breast milk Hg and average maternal hair Hg with breast milk Hg, respectively. The regression coefficients were in the range of 0.19 to 0.90, and P values were in the range of 0.0001 to 0.1490. Further recommendations include fish advisories to prevent critical Hg exposures during reproductive life and investigation of neurobehavioral performance of this study population.

Organochlorine contaminants and biomarker response in double-crested cormorants nesting in Green Bay and Lake Michigan, Wisconsin, USA.

Double-crested cormorant (Phalacrocorax auritus) eggs at pipping and sibling 10-day-old chicks were collected from two colonies in Green Bay, WI, one colony in Lake Michigan, WI, and reference colonies in South Dakota and Minnesota. Egg contents and chicks were analyzed for organochlorine contaminants including polychlorinated biphenyl (PCB) congeners. Livers of embryos and chicks were assayed for hepatic microsomal ethoxyresorufin-O-dealkylase (EROD) activity. Eggshell thickness and the physical dimensions of embryo brains were measured. Concentrations of organochlorines, including p,p'-DDE (p,p'-dichlorodiphenyldichloroethylene), PCBs, and PCB congeners were generally an order of magnitude higher in eggs and chicks from Wisconsin than from reference locations. Total PCBs averaged 10-13 microg/g wet weight in eggs from three Wisconsin colonies compared to 0.9 microg/g PCBs from reference locations. Double-crested cormorant chicks accumulated on average 33-66 microg PCBs/day and 7-12 microg p,p'-DDE/day in the Wisconsin colonies compared to 0 microg PCBs/day and 1 microg p,p'-DDE/day in the reference colonies. At pipping, EROD activity in the livers of cormorant embryos was significantly higher in the Wisconsin colonies and significantly correlated with PCBs and the toxic equivalents (TEQs) of aryl hydrocarbon-active PCB congeners relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin. However, in 10-day-old chicks EROD activity was not consistently different among colonies and was not correlated with PCBs or TEQs. A significant negative relationship between embryo brain asymmetry and the size of the egg suggested that physical constraint might be an important factor influencing the response of this bioindicator. Thinner eggshells in two colonies located near Door County, Wisconsin, suggested that historic p,p'-DDE residues associated with orchards are still an important source of p,p'-DDE in the local environment.