Lethal and sublethal toxicity of perfluorooctanoic acid (PFOA) in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow).

Perfluoroalkyl substances (PFAS), including perfluorooctanoic acid (PFOA), are industrial chemicals that are of concern due to their environmental presence, persistence, bioaccumulative potential, toxicity, and capacity for long-range transport. Despite a large body of research on environmental exposure, insufficient chronic aquatic toxicity data exist to develop water quality targets for clean-up of federal contaminated sites in Canada. Thus, our objective was to assess the aqueous toxicity of PFOA in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow). Toxicity data were analyzed based on measured PFOA concentrations. Amphipod exposures were 42 d (0.84-97 mg/L) and examined survival, growth, and reproduction. Fathead minnow exposures were 21 d (0.010-76 mg/L), which encompassed hatching (5 d) and larval stages until 16 d post-hatch; endpoints included hatching success, deformities at hatch, and larval survival and growth. Amphipod survival was significantly reduced at 97 mg/L (42-d LC50 = 51 mg/L), but growth and reproduction were more sensitive endpoints (42-d EC50 for both endpoints = 2.3 mg/L). Fathead minnows were less sensitive than amphipods, exhibiting no significant effects in all endpoints with the exception of uninflated swim bladder, which was significantly higher at 76 mg/L (15%) than controls (0%). Maximum concentrations of PFOA are generally in the ng/L range in global surface waters, but can reach the µg/L range in close proximity to major source inputs; therefore, environmental concentrations are well below those that caused toxicity in the current study. Our data will provide valuable information with which to assess the risk of PFOA at contaminated sites, and to set a target for site remediation.

Increased Temperature and Turbulence Alter the Effects of Leachates from Tire Particles on Fathead Minnow (Pimephales promelas).

Tire particles are of concern as a stressor due to the combination of their chemical constituents, high emission rates, and global distribution. Once in the environment, they will interact with physical parameters (e.g., UV, temperature). The interaction of chemical pollution with changing physical environmental parameters is often underestimated in ecotoxicology. Here, we investigate the role of temperature, mechanical stress (i.e., turbulence), UV, and CO2 on the effects of tire leachates on fish. Two samples of tire particles were exposed to four different levels of each physical stressor. A toxicological assessment was performed with fathead minnow embryos assessing five end points (hatching success, time to hatch, length, deformities, and heart rate). Results showed that variations of temperature and mechanical stress affect the toxicological impact of tire leachates. Zn and/or polycyclic aromatic hydrocarbons (pyrene, phenanthrene, chrysene, benzo[a]pyrene, anthracene, naphthalene, fluoranthene, and benzo[ghi]perylene) were identified in the leachate and tire samples by Raman/surface-enhanced Raman spectroscopy and gas chromatography with mass spectroscopy, respectively.

Chronic Embryo-Larval Exposure of Fathead Minnows to the Pharmaceutical Drug Metformin: Survival, Growth, and Microbiome Responses.

Metformin is a glucose-lowering drug commonly found in municipal wastewater effluents (MWWEs). The present study investigated the chronic effects of metformin in early-life stages of the fathead minnow (Pimephales promelas). Endpoints assessed were growth, survival, and deformities. The larval gut microbiome was also examined using 16 S ribosomal RNA gene amplicon sequencing to determine microbial community composition and alpha and beta diversity. Eggs and larvae were exposed to metformin measured concentrations (mean [standard deviation]) of 0.020 (0.017) µg/L (for controls) and 3.44 (0.23), 33.6 (1.6), and 269 (11) µg/L in a daily static-renewal setup, with 20 embryos per beaker. The low and middle metformin exposure concentrations represent river and MWWE concentrations of metformin. To detect small changes in growth, we used 18 replicate beakers for controls and 9 replicates for each metformin treatment. Over the 21-d exposure (5 d as embryos and 16 d posthatch [dph]), metformin did not affect survival or growth of larval fish. Hatch success, time to hatch, deformities in hatched fry, and survival were similar across all treatments. Growth (wet wt, length, and condition factor) assessed at 9 and 16 dph was also unaffected by metformin. Assessment of the microbiome showed that the larvae microbiome was dominant in Proteobacteria and Firmicutes, with small increases in Proteobacteria and decreases in Firmicutes with increasing exposure to metformin. No treatment effects were found for microbiome diversity measures. Control fish euthanized with the anesthetic tricaine methane sulfonate had decreased alpha diversity compared to those sampled by spinal severance. This experiment demonstrates that metformin at environmentally relevant concentrations (3.44 and 33.6 µg/L) and at 10 times MWWE concentrations (269 µg/L) does not adversely affect larval growth or gut microbiome in this ubiquitous freshwater fish species. Environ Toxicol Chem 2022;41:635-647. © 2021 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

A Deep Dive into the Complex Chemical Mixture and Toxicity of Tire Wear Particle Leachate in Fathead Minnow.

The ecological impact of tire wear particles in aquatic ecosystems is a growing environmental concern. We combined toxicity testing, using fathead minnow (Pimephales promelas) embryos, with nontarget high-resolution liquid chromatography Orbitrap mass spectrometry to characterize the toxicity and chemical mixture of organic chemicals associated with tire particle leachates. We assessed: 1) exposure to tire particle leachates after leaching for 1-, 3-, and 10-d; and 2) the effect of the presence and absence of small tire particulates in the leachates. We observed a decrease in embryonic heart rates, hatching success, and lengths, as well as an increase in the number of embryos with severe deformities and diminished eye and body pigmentation, after exposure to the leachates. Overall, there was a pattern whereby we observed more toxicity in the 10-d leachates, and greater toxicity in unfiltered leachates. Redundancy analysis showed that several benzothiazoles and aryl-amines were correlated with the toxic effects observed in the embryos. These included benzothiazole, 2-aminobenzothiazole, 2-mercaptobenzothiazole, N,N'-diphenylguanidine, and N,N'-diphenylurea. However, many other chemicals characterized as unknowns are likely to also play a key role in the adverse effects observed. Our study provides insight into the types of chemicals likely to be important toxicological drivers in tire leachates, and improves our understanding of the ecotoxicological impacts of tire wear particles. Environ Toxicol Chem 2022;41:1144-1153. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The acute toxicity of bitumen-influenced groundwaters from the oil sands region to aquatic organisms.

The extraction of surface mined bitumen from oil sands deposits in northern Alberta, Canada produces large quantities of liquid tailings waste, termed oil sands process-affected water (OSPW), which are stored in large tailings ponds. OSPW-derived chemicals from several tailings ponds migrating past containment structures and through groundwater systems pose a concern for surface water contamination. The present study investigated the toxicity of groundwater from near-field sites adjacent to a tailings pond with OPSW influence and far-field sites with only natural oil sands bitumen influence. The acute toxicity of unfractionated groundwater and isolated organic fractions was assessed using a suite of aquatic organisms (Pimephales promelas, Oryzias latipes, Daphnia magna, Hyalella azteca, Lampsilis spp., Ceriodaphnia dubia, Hexagenia spp., and Vibrio fischeri). Assessment of unfractionated groundwater demonstrated toxicity towards all invertebrates in at least one far-field sample, with both near-field and far-field samples with bitumen influence toxic towards P. promelas, while no toxicity was observed for O. latipes. When assessing the unfractionated groundwater and isolated organic fractions from near-field and far-field groundwater sites, P. promelas and H. azteca were the most sensitive to organic components, while D. magna and L. cardium were most sensitive to the inorganic components. Groundwater containing appreciable amounts of dissolved organics exhibited similar toxicities to sensitive species regardless of an OSPW or natural bitumen source. The lack of a clear distinction in relative acute toxicities between near-field and far-field samples indicates that the water-soluble chemicals associated with bitumen are acutely toxic to several aquatic organisms. This result, combined with the similarities in chemical profiles between bitumen-influenced groundwater originating from OSPW and/or natural sources, suggests that the industrial bitumen extraction processes corresponding to the tailings pond in this study are not contributing unique toxic substances to groundwater, relative to natural bitumen compounds present in groundwater flow systems.

Toxicity of sediment-associated substituted phenylamine antioxidants on the early life stages of Pimephales promelas and a characterization of effects on freshwater organisms.

Substituted phenylamine antioxidants (SPAs) are high production volume chemicals that are incorporated into a variety of commercial products (e.g., polymers, dyes, lubricants). There are few data on chronic toxicity of SPAs to fish and no data on the toxicity of SPAs to the early life stages of fish. The physicochemical properties of SPAs would suggest that if they were to enter an aquatic ecosystem they would partition into sediment. Therefore, the present study focused on investigating the chronic effect of sediment-associated SPAs to the early life stages of the fathead minnow (Pimephales promelas). Eggs and larvae were exposed to sediment spiked with diphenylamine (DPA), N-phenyl-1-napthylamine (PNA), N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA), or 4,4'-methylene-bis[N-sec-butylaniline] (MBA). The most sensitive endpoint for DPA, PNA, and DPPDA was total survival with 21-d median lethal concentrations (LC50s) based on concentration in overlying water of 1920, 74, and 35 µg/L, respectively. The most sensitive endpoint for MBA was growth with a 21-d median effective concentration (EC50) of 71 µg/L. The same endpoints were the most sensitive in terms of concentrations of DPA, PNA, DPPDA, and MBA in sediment (101, 54, 111, and 76 µg/g dry wt, respectively). Species sensitivity distributions (SSDs) were constructed for each SPA based on acute and chronic toxicity data generated in the present study and found in the literature. Overall, P. promelas was in the midrange of chronic sensitivity, with the most sensitive species being Tubifex tubifex. The SSDs indicate that DPA based on concentration in water is the least toxic to aquatic biota of the 4 SPAs investigated. The constructed SSDs indicate that a concentration in water and sediment of 1 µg/L and 1 µg/g dry weight, respectively, would be protective of >95% of the aquatic species tested. Environ Toxicol Chem 2017;36:2730-2738. © 2017 SETAC.

Effects of multi-well plate incubation on embryo-larval development in the fathead minnow (Pimephales promelas).

Fathead minnow embryos and larvae are frequently used in toxicology, including short-term embryo-only tests which often use small volumes of test solution. The effect that such conditions may have on fathead minnow development has yet to be explicitly described. Here we compared rates of embryonic development in fathead minnow embryos reared under standard light and temperature conditions with a range of possible methods. All methods yielded excellent control survival. We demonstrated that fathead minnow embryos incubated in a range of small volumes in multi-well plates (500 µL to 2 mL per embryo) did not substantially vary in developmental rate, but flexed less frequently as embryos, hatched smaller, later and with larger yolk-sacs, and initiated feeding later than embryos reared in an excess of solution (20 mL per embryo) with or without supplemental aeration. Faster hatch and growth were promoted with an orbital shaker, but growth benefits were not sustained into the larval stage. Developmental differences persisted in larvae reared to 20 days post-fertilization when monitoring ceased, but growth differences did not magnify and in some measurements partially resolved. To our knowledge we are the first to report effects of incubation in multi-well plates in any fish taxa. As our data revealed that the eleutheroembryonic stage for fathead minnow may be prolonged in multi-well plates, this may allow the use of longer toxicity tests using fathead minnow embryos without conflicting with existing animal welfare legislation in many countries.