Influence of Lipophilicity on the Toxicity of Bisphenol A and Phthalates to Aquatic Organisms.

Bisphenol A (BPA) and phthalates are among the most popular plasticizers used today and have been reported ubiquitously in surface water, ground water, and sediment. For aquatic organisms, BPA was the most toxic (96 h LC50s) to aquatic invertebrates (0.96-2.70 mg/L) and less toxic to fish (6.8-17.9 mg/L). The toxicity of BPA to amphibians differed among developmental stages, with embryos having an LC50 of 4.6-6.8 mg/L and juveniles 0.50-1.4 mg/L. The toxicity of phthalates is affected by aromatic ring substitution, alkyl chain length, and metabolism. The toxicity (96 h LC50s) of phthalates was similar to aquatic invertebrates (0.46-377 mg/L) and fish (0.48-121 mg/L). In general, the toxicity of phthalates appears to be highest around a log KOW of 6, which corresponds to the highest potential for bioconcentration and bioaccumulation. In conclusion, the lipophilicity of BPA and phthalates influence their toxicity to aquatic species.

Lethal and sublethal effects of phthalate diesters in Silurana tropicalis larvae.

Phthalates are compounds used in polymers to increase their flexibility and are now ubiquitous in the environment as a result of widespread use. Because few studies have focused on the adverse effects of these chemicals in aquatic species, the present study aimed to determine the effects of phthalate diesters in amphibians. Western clawed frog (Silurana tropicalis) tadpoles were acutely exposed to water spiked with monomethyl phthalate (MMP; 1.3-1595.5 mg/L), dimethyl phthalate (DMP; 0.03-924.0 mg/L), or dicyclohexyl phthalate (DCHP; 0.3-99.3 mg/L). Because few studies have addressed the toxicity of these specific phthalates in most organisms, the present study used higher concentrations of these chemicals to determine their toxicity pathways in amphibians and at the same time investigate a suite of genes known to be altered by the well-studied phthalates. Both DMP and DCHP increased larval mortality (9.1-924.0 mg/L DMP and 4.1-99.3 mg/L DCHP), increased frequency of malformations in tadpoles (0.1-34.1 mg/L DMP and 4.1-19.0 mg/L DCHP), and up-regulated cellular stress-related messenger-RNA (mRNA) levels (4.1 mg/L DCHP). To characterize the molecular toxicity pathway of these phthalates in tadpoles, transcriptome analysis was conducted using a custom microarray. Parametric analysis of gene set enrichment revealed important changes in the expression of genes related to drug metabolism and transport, liver metabolism, xenobiotic clearance, and xenobiotic metabolism after DMP and DCHP treatments, although these responses were less pronounced with MMP (the metabolite of DMP). The present study is one of the few studies that demonstrated complementarity between gene expression analysis and organismal effects. Environ Toxicol Chem 2016;35:2511-2522. © 2016 SETAC.

Genotoxic and carcinogenic products arising from reductive transformations of the azo dye, Disperse Yellow 7.

Selected aromatic azo and benzidine based dyes are priority compounds under the Government of Canada's Chemical Management Plan (CMP) for environmental risk assessments. Organic compounds undergo chemical and biological transformations when they interact with environmental matrices and biotic species; identifying the transformation products is thus a critical component of the risk assessment process. Here, we used zero valent iron (ZVI) to initiate the reduction of the diazo compound dye Disperse Yellow 7 (DY 7). Using state-of-the-art accurate mass Liquid Chromatography-Quadrupole Time of Flight-Mass Spectroscopy (LC-QToF-MS), four transformation products were conclusively identified, while a fifth product was tentatively ascertained. The conclusively established transformation products included p-phenylenediamine (p-PDA, a known genotoxin), 4-aminoazobenzene (4-AAB, a category 2 carcinogen) and 4-aminobiphenyl (4-ABP, a category 1 human carcinogen). 4-ABP is thought to form via a benzidine rearrangement; this is the first report of DY 7 undergoing a benzidine rearrangement. Given the importance of reduction processes in the metabolism of organic contaminants by aquatic species, we used LC-MS/MS to analyze sediment samples that had been generated previously upon exposure of Western clawed frogs (Silurana tropicalis) to DY 7 (at exposure levels where cellular stress was observed in S. tropicalis). We found p-PDA, 4-AAB, and 4-ABP were present in all exposures, but not in any of the sediment controls, demonstrating that upon release of DY 7 to the aquatic environment, sediment dwelling organisms will metabolize DY 7 to generate known (and suspected) human carcinogens, including through a previously unreported in vivo benzidine rearrangement to produce 4-ABP.

Chronic exposures to monomethyl phthalate in Western clawed frogs.

Polymer flexibility and elasticity is enhanced by plasticizers. However, plasticizers are often not covalently bound to plastics and thus can leach from products into the environment. Much research effort has focused on their effects in mammalian species, but data on aquatic species are scarce. In this study, Western clawed frog (Silurana tropicalis) embryos were exposed to 1.3, 12.3, and 128.7mg/L monomethyl phthalate (MMP) until the juvenile stage (11weeks) and to 1.3mg/L MMP until the adult stage (51weeks). MMP decreased survival, hastened metamorphosis, and biased the sex ratio toward males (2M:1F) at the juvenile stage without altering the expression of a subset of thyroid hormone-, sex steroid-, cellular stress- or transcription regulation-related genes in the juvenile frog livers. At the adult stage, exposure to MMP did not have significant adverse health effects, except that females had larger interocular distance and the expression of the heat shock protein 70 was decreased by 60% in the adult liver. In conclusion, this study shows that MMP is unlikely to threaten amphibian populations as only concentrations four orders of magnitude higher than the reported environmental concentrations altered the animal physiology. This is the first complete investigation of the effects of phthalates in a frog species, encompassing the entire life cycle of the organisms.

Plasticizer endocrine disruption: Highlighting developmental and reproductive effects in mammals and non-mammalian aquatic species.

Due to their versatility, robustness, and low production costs, plastics are used in a wide variety of applications. Plasticizers are mixed with polymers to increase flexibility of plastics. However, plasticizers are not covalently bound to plastics, and thus leach from products into the environment. Several studies have reported that two common plasticizers, bisphenol A (BPA) and phthalates, induce adverse health effects in vertebrates; however few studies have addressed their toxicity to non-mammalian species. The aim of this review is to compare the effects of plasticizers in animals, with a focus on aquatic species. In summary, we identified three main chains of events that occur in animals exposed to BPA and phthalates. Firstly, plasticizers affect development by altering both the thyroid hormone and growth hormone axes. Secondly, these chemicals interfere with reproduction by decreasing cholesterol transport through the mitochondrial membrane, leading to reduced steroidogenesis. Lastly, exposure to plasticizers leads to the activation of peroxisome proliferator-activated receptors, the increase of fatty acid oxidation, and the reduction in the ability to cope with the augmented oxidative stress leading to reproductive organ malformations, reproductive defects, and decreased fertility.

Sediment contaminated with the Azo Dye disperse yellow 7 alters cellular stress- and androgen-related transcription in Silurana tropicalis larvae.

Azo dyes are the most commonly used type of dye, accounting for 60-70% of all organic dye production worldwide. They are used as direct dyes in the textile, leather, printing ink, and cosmetic industries. The aim of this study was to assess the lethal and sublethal effects of the disazo dye Disperse Yellow 7 (DY7) in frogs to address a knowledge gap regarding mechanisms of toxicity and the potential for endocrine disrupting properties. Larvae of Silurana tropicalis (Western clawed frog) were exposed to DY7-contaminated water (0 to 22 µg/L) and sediment (0 to 209 µg/g) during early larval development. The concentrations used included the range of similar azo dyes found in surface waters in Canada. A significant decrease in tadpole survivorship was observed at 209 µg/g while there was a significant increase in malformations at the two highest concentrations tested in sediment. In the 209 µg/g treatment, DY7 significantly induced hsp70 (2.5-fold) and hsp90 (2.4-fold) mRNA levels, suggesting that cells required oxidative protection. The same treatment also altered the expression of two androgen-related genes: decreased ar (2-fold) and increased srd5a2 (2.6-fold). Furthermore, transcriptomics generated new hypotheses regarding the mechanisms of toxic action of DY7. Gene network analysis revealed that high concentrations of DY7 in sediment induced cellular stress-related gene transcription and affected genes associated with necrotic cell death, chromosome condensation, and mRNA processing. This study is the first to report on sublethal end points for azo dyes in amphibians, a growing environmental pollutant of concern for aquatic species.

Toxicity of the azo dyes Acid Red 97 and Bismarck Brown Y to Western clawed frog (Silurana tropicalis).

Azo compounds are used in a variety of industrial applications, such as textile colorant. Azo dyes have been found to contaminate aquatic environments and it has been shown that these compounds could potentially be toxic or induce endocrine disruption in aquatic organisms. However, there are few data available on the toxicity of these dyes, specifically Acid Red 97 (AR97) and Bismarck Brown Y (BBY). The aim of this study was to determine the toxicity and the endocrine-disrupting properties of AR97 and BBY in frogs. As fugacity modeling predicted that both compounds would sorb to sediment, sediment exposures were performed using a geometric range of concentrations (0, 1, 10, 100 and 1,000 ppm). Both AR97 and BBY dyes were not lethal to Silurana tropicalis embryos; however, BBY significantly induced malformations. Gene expression analysis of oxidative stress and mutagen-related genes was performed in BBY-treated larvae. There were significant two-fold increases of the tumor-suppressing protein p53 and heat shock protein 70 mRNA at 1,000 ppm suggesting that BBY induces cellular stress in early S. tropicalis development. Transcripts of the heat shock protein 90 did not change. Furthermore, reproductive-related genes were assessed and a 2.1-fold change was observed in the mRNA of the steroidogenic acute regulatory protein while steroid 5 alpha-reductase type 2 and androgen receptor transcript levels did not vary among treatments. In conclusion, high concentrations of BBY lead to increased developmental defects in frog embryogenesis and early larval development.