Sex-Specific Differences of Steroid Receptors Following Exposure to Environmentally Relevant Concentrations of Phenothiazine in Fundulus heteroclitus.

Phenothiazine (PTZ) is a heterocyclic thiazine compound used for industrial and medical purposes. Through environmental surveillance studies, PTZ was found being discharged into a local river in Connecticut. Phenothiazine has been shown to act similarly to endocrine disrupting chemicals. This study sought to identify sex specific hormone receptor changes in Fundulus heteroclitus in response to PTZ exposure. Fundulus heteroclitus, also known as mummichog, are small fish native to the Atlantic coast of the United States and Canada. They reside in brackish waters and can survive harsh toxic environments. This model organism is native to the polluted waters found in Connecticut. In this study, fish were exposed to PTZ concentrations of 0.5 ppm, 1.0 ppm, and 2.0 ppm for 1 week. Following exposure, brain, liver, and gonad tissues were harvested; cDNA was synthesized; and mRNA expression was assessed for 6 different hormone receptors. Compared with vehicle control (ethanol) differences in mRNA expression, levels of hormone receptors were observed in various tissues from male and female fish. Many of the tissues assessed showed changes in expression level, while only female liver and testis showed no change. These results implicate PTZ as a potential endocrine disrupting compound to mummichog at environmentally relevant concentrations.

Chronic exposure to benzyl butyl phthalate (BBP) alters social interaction and fear conditioning in male adult rats: alterations in amygdalar MeCP2, ERK1/2 and ERα.

OBJECTIVES: Benzyl Butyl Phthalate (BBP) is an industrial plasticizer that has an unknown action in the central nervous system. Phthalates have recently been associated with behavioral actions that are linked to their endocrine disrupting properties. The purpose of this study was to investigate the behavioral and molecular effects of BBP treatment in male rats. DESIGN: Male rats were chronically exposed to BBP in the drinking water (5.0 ppm and 10.0 ppm) throughout adolescence and into the adult phase of life. Their behavior was then assessed in a learning and memory task (fear conditioning), open field exploration and a test of sociability. RESULTS: BBP treated rats showed decreased freezing in fear conditioning, no changes in open field exploration, and increased aberrant social behavior. Rats were sacrificed at post natal day 140 and blood and brains were harvested and processed. We found increased hormonally active estrogen, 17-ß estradiol, in the serum of BBP treated rats. BBP treatment also induced changes in amygdalar proteins related to synaptic plasticity including decreased MeCP2 levels that correlated with tests of sociability with no changes in stress related proteins such as nuclear factor kappa B (NFkB). We also found alterations in physiological responses as measured by body weight without changes in food consumption suggesting disruption of metabolism and body homeostasis. CONCLUSIONS: We suggest that BBP administration disrupts normal learning and social behavior, and that these effects could be related to alterations of amygdala function.

Impact of benzyl butyl phthalate on shoaling behavior in Fundulus heteroclitus (mummichog) populations.

Fundulus heteroclitus preference for association with familiar conspecifics of similar body length was impacted by benzyl butyl phthalate (BBP); this was found to be a statically significant result with a p < 0.0001. When presented with equally sized shoals consisting of either large or small fish, the majority of unexposed (84%) and acetone exposed control (82%) fish selected the shoal of large fish. A small number of control fish chose either the shoal of small fish (6% and 10%) or the neutral zone (10% and 8%) where they were clear morphological outliers. Fish exposed to 0.1 mg/L BBP exposure daily for four weeks selected the shoal of small fish more often than unexposed or acetone controls (7.5- and 4.5-fold respectively). They also remained in the neutral zone and displayed agitation at levels more than twice that of control. Agitation and shoal choice disruption are quantifiable behavioral responses that support the use of F. heteroclitus as a model for detecting sub-lethal BBP exposure.

Pesticides and Alcohol in Imported and Domestic Wine Sold in Connecticut.

Pesticides not allowed for use in the United States may be applied in foreign countries on grapes destined for wine production. There is concern that imported wine may contain residues of these pesticides. Of 51 wines tested from five foreign countries, 17 samples contained residues of Procymidone, a fungicide not allowed on crops in the United States. Three of 18 wines tested contained residues of Carbaryl at concentrations below the allowable tolerance for grapes. Twelve wines from the United States contained no detectable residues of Procymidone. The alcohol content of all wines met label claims.

The Formation of N-Nitrosodimethylamine and N-Nitrodimethylamine from the Plant Growth Regulator Daminozide (Alar).

Daminozide (Alar) was found to produce N-nitrosodimethylamine (NDMA) and N-nitrodimethylamine (NITDMA) when treated with nitrite in acidic solution. Unsymmetrical dimethylhydrazine (UDMH), the hydrolysis product of Daminozide, gave the same products. Use of 15N-nitrite and 15N-Daminozide showed that both the nitroso group of NDMA and the nitro group of NITDMA arise from the nitrite.

Pesticides in Orange Juice Sold in Connecticut.

There is concern that orange juice from foreign countries may contain residues of pesticides not allowed in the United States. Of 17 orange juices examined, 15 listed Brazil as the source of all or part of the juice used. Six samples contained residues. All pesticides found were allowed for use in the United States, and all residues were well below EPA allowable tolerances in oranges.

Herbicides in Pooled Raw Milk in Connecticut.

Pooled raw milk samples collected in Connecticut were analyzed for the herbicides 2,4-D, Atrazine, Simazine, and Alachlor. For 2,4-D, 25.6% of the samples had no detectable level, for Atrazine 7.7%, and for Alachlor 10.3%. None contained Simazine. Concentrations averaged 0.021 ppm for 2,4-D, 0.0075 for Atrazine, and 0.0067 for Alachlor. No sample exceeded the EPA tolerance for any of the herbicides. Analysis was by enzyme immunoassay.