Hydraulic fracturing fluid are complex mixtures containing high concentrations of salts (up to 330,000 ppm), organic, and metal contaminants. However, little data exist on the potential mechanisms of toxicity of these flowback and produced wastewaters (FPW) on aquatic biota. Juvenile rainbow trout were exposed to either control, FPW (2.5 or 7.5%), FPW that had been treated with activated charcoal (AC), or a custom salt-matched control (SW; replicating only the salt content of FPW) for 48 hours. Gill histology revealed decreases in interlamellar cell mass (ILCM) and mean lamellar length in all treatments (FPW, AC and SW) compared to control, indicative of hyperosmotic stress. Liver CYP1A1 activity was significantly elevated by 7.5-fold in the FPW 7.5% treatment only, indicative of Phase I metabolism. Superoxide dismutase activity significantly decreased in the gills to all treatments with the lowest activity occurring in the 7.5% FPW group. Catalase activity increased in liver with the highest values noted in fish exposed to 7.5% FPW. No changes were observed with respect to glutathione-S-transferase, while increased lipid peroxidation was only observed in both FPW treatments (2.5, 7.5%). These data suggest a characteristic signature of FPW impact which may help in risk assessment and biomonitoring of FPW spills.
Hagfish consume carrion, potentially exposing them to hypoxia, hypercapnia, and high environmental ammonia (HEA). We investigated branchial and cutaneous ammonia handling strategies by which Pacific hagfish () tolerate and recover from high ammonia loading. Hagfish were exposed to HEA (20 mmol/l) for 48 h to elevate plasma total ammonia (T) levels before placement into divided chambers for a 4-h recovery period in ammonia-free seawater where ammonia excretion () was measured independently in the anterior and posterior compartments. Localized HEA exposures were also conducted by subjecting hagfish to HEA in either the anterior or posterior compartments. During recovery, HEA-exposed animals increased in both compartments, with the posterior compartment comprising ~20% of the total compared with ~11% in non-HEA-exposed fish. Plasma T increased substantially when whole hagfish and the posterior regions were exposed to HEA. Alternatively, plasma T did not elevate after anterior localized HEA exposure. was concentration dependent (0.05-5 mmol/l) across excised skin patches at up to eightfold greater rates than in skin sections that were excised from HEA-exposed hagfish. Skin excised from more posterior regions displayed greater than those from more anterior regions. Immunohistochemistry with hagfish-specific anti-rhesus glycoprotein type c (α-hRhcg; ammonia transporter) antibody was characterized by staining on the basal aspect of hagfish epidermis while Western blotting demonstrated greater expression of Rhcg in more posterior skin sections. We conclude that cutaneous Rhcg proteins are involved in cutaneous ammonia excretion by Pacific hagfish and that this mechanism could be particularly important during feeding.
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