Greg G. Pyle

Metal-PAH mixtures in the aquatic environment: A review of co-toxic mechanisms leading to more-than-additive outcomes

Mixtures of metals and polycyclic aromatic hydrocarbons (PAHs) occur ubiquitously in aquatic environments, yet relatively little is known regarding their combined toxicities. Emerging reports investigating the additive mortality in metal-PAH mixtures have indicated that more-than-additive effects are equally as common as strictly-additive effects, raising concern for ecological risk assessment typically based on the summation of individual toxicities. Moreover, the current separation of focus between in vivo and in vitro studies, and fine- and coarse-scale endpoints, creates uncertainty regarding the mechanisms of co-toxicity involved in more-than-additive effects on whole organisms. Drawing from literature on metal and PAH toxicity in bacteria, protozoa, invertebrates, fish, and mammalian models, this review outlines several key mechanistic interactions likely to promote more-than-additive toxicity in metal-PAH mixtures. Namely, the deleterious effects of PAHs on membrane integrity and permeability to metals, the potential for metal-PAH complexation, the inhibitory nature of metals to the detoxification of PAHs via the cytochrome P450 pathway, the inhibitory nature of PAHs towards the detoxification of metals via metallothionein, and the potentiated production of reactive oxygenated species (ROS) in certain metal (e.g. Cu) and PAH (e.g., phenanthrenequinone) mixtures. Moreover, the mutual inhibition of detoxification suggests the possibility of positive feedback among these mechanisms. The individual toxicities and interactive aspects of contaminant transport, detoxification, and the production of ROS are herein discussed.

THE INFLUENCE OF WATER HARDNESS, pH, AND SUSPENDED SOLIDS ON NICKEL TOXICITY TO LARVAL FATHEAD MINNOWS (PIMEPHALES PROMELAS)

Nickel (Ni) is an ubiquitous, naturally occurring metalthat is associated with metal mining and other industrialactivities. Despite elevated Ni concentrations reportedfor many industrial receiving waters, Ni receives littleresearch attention addressing factors influencing itstoxicity to freshwater fish. This study examined theinfluence of water hardness, pH, and total suspended solids(TSS) in soft, reconstituted water on Ni toxicity to larvalfathead minnows (Pimephales promelas). Increasingwater hardness from 20 to 140 mg L-1 (as CaCO3) reduced acute Ni toxicity by 5-fold (96-h LC50s 0.45 and 2.27 mg Ni L-1, respectively). Low pH had a slight protective effect against Ni toxicity relative to neutral pH conditions. At pH 5.5, the 96-h LC50 was 0.69 mg Ni L-1, compared to 0.54 mg Ni L-1 at pH 7.0. However,Ni toxicity was significantly reduced at pH 8.5 where the 96-h LC50 was 2.21 mg Ni L-1. These results were explainedon the basis of Ni speciation. Total suspended solids also reduced Ni toxicity (expressed as 96-h LC50s) from 0.35 to 1.12 mg Ni L-1 over a TSS range of 10 to 100 mg L-1.This reduction of toxicity due to TSS is significant becausemine effluents often have a combination of elevated TSS andmetals. The ameliorative effect of TSS was not as significantas high hardness or pH probably because there is a TSS threshold, after which physical irritation to fish gills counteracts any protective effect conferred by TSS. This finding is relevant to choices made in design of mine effluenttreatment systems; i.e., there may be an optimum range ofTSS concentrations that protect aquatic biota againsteffects of metals that remain after treatment.

Copper Binding Dynamics and Olfactory Impairment in Fathead Minnows (Pimephales promelas)

When fish are exposed to sublethal, environmentally relevant Cu concentrations, olfactory acuity is impaired. The goals of the present study were to investigate the binding dynamics of waterborne Cu in the olfactory epithelium (OE), to examine the influence of calcium (Ca(2+)) on Cu binding, and to link Cu-OE binding to changes in olfactory acuity. Using short-term in vivo waterborne exposures to (64)Cu, we found that Cu accumulates rapidly in the OE, reaching a plateau by 3 h. The binding affinity (log K(Cu-OE)) and binding capacity (B(max)) of (64)Cu in the OE were 6.7 and 10.0 nmol Cu g(-1), respectively. As waterborne Ca(2+) was increased from 50 to 1000 microM L(-1), the B(max) of Cu decreased by approximately 50% while the log K(Cu-OE) remained constant, indicative of noncompetitive inhibition. Using electro-olfactograms (EOG), short-term exposures to 160 and 240 nmol Cu L(-1) were found to reduce olfactory responses to 10(-5) M l-arginine by 72 and 79%, respectively. Short-term exposure to 160 nmol Cu L(-1) also caused a 15-fold reduction in behavioral responses to a food stimulus. Interestingly, increasing waterborne Ca(2+) did not reduce the effects of Cu on EOG or behavioral responses. These results demonstrate that short-term, environmentally realistic concentrations of Cu not only bind to the OE of fathead minnows but also impair their olfactory sensitivity and behavioral responses to olfactory stimuli. Waterborne Ca(2+) reduces Cu-OE binding but does not protect against olfactory impairment.

Do you smell what I smell? Olfactory impairment in wild yellow perch from metal-contaminated waters.

In this study, we sampled yellow perch from three lakes along a metal-contamination gradient and examined their olfactory ability in response to conspecific chemical alarm cues and metal-binding characteristics of their olfactory epithelium (OE). We measured the electrophysiological response at the OE, tested their antipredator behaviour and measured neuronal density at the olfactory rosette and bulb. Yellow perch from contaminated lakes exhibited significantly larger electrophysiological responses to alarm cues than clean lake fish, but showed no antipredator behaviour contrary to clean lake fish. Neuron density did not differ at either the olfactory rosette or bulb between clean and contaminated fish. Unlike fishes raised under laboratory or aquaculture settings, fish from contaminated lakes possessed a functional OE after metal exposure, but similar to laboratory/aquaculture fishes, yellow perch did not exhibit olfactory-mediated behaviours. Thus, wild fish from contaminated lakes can detect chemical stimuli but olfactory signal processing is disrupted which could alter ecological functioning.

Contaminant-specific targeting of olfactory sensory neuron classes: Connecting neuron class impairment with behavioural deficits

The olfactory system of fish comprises several classes of olfactory sensory neurons (OSNs). The odourants L-alanine and taurocholic acid (TCA) specifically activate microvillous or ciliated OSNs, respectively, in fish. We recorded electro-olfactograms (EOG) in fathead minnows (Pimephales promelas; a laboratory-reared model species) and wild yellow perch (Perca flavescens) whose olfactory chambers were perfused with either L-alanine or TCA to determine if OSN classes were differentially vulnerable to contaminants, in this case copper or nickel. Results were consistent in both species and demonstrated that nickel targeted and impaired microvillous OSN function, while copper targeted and impaired ciliated OSN function. This result suggests that contaminant-specific effects observed in model laboratory species extrapolate to wild fish populations. Moreover, fathead minnows exposed to copper failed to perceive a conspecific alarm cue in a choice maze, whereas those exposed to nickel could respond to the same conspecific cue. These results demonstrate that fathead minnows perceive conspecific, damage-released alarm cue by ciliated, but not microvillous, OSNs. Fish living in copper-contaminated environments may be more vulnerable to predation than those in clean lakes owing to targeted effects on ciliated OSNs.

Radionuclide equilibria between the aquatic environment and fish tissues

Abstract Equilibrium conditions are often assumed in dose and risk calculations based on the simple linear concentration factor model, a commonly used model in the study of contaminant flow through ecosystems. It has been argued that by using a power function model to describe radionuclide retention in fish, equilibrium may never be achieved under natural conditions, thereby violating the equilibrium requirement in the concentration factor model. Our results demonstrate uranium-series radionuclide equilibria in a natural population of common white sucker (Catostomus commersoni). Concentration factors indicated that 226 Ra, 232 Th, 230 Th, and 228 Th preferentially deposited in bone over muscle. Although 226 Ra had the highest concentration in bone, 228 Th yielded the highest concentration factors for water-to-bone transfer. 232 Th may not be in equilibrium because of a growth dilution effect.

Olfactory recovery of wild yellow perch from metal contaminated lakes.

Fish depend on their sense of smell for a wide range of vital life processes including finding food, avoiding predators and reproduction. Various contaminants, including metals, can disrupt recognition of chemical information in fish at very low concentrations. Numerous studies have investigated metal effects on fish olfaction under controlled laboratory conditions. However, few have measured olfactory acuity using wild fish in source water. In this study, we used electro-olfactography (EOG) to measure the olfactory acuity of wild yellow perch (Perca flavescens) from a clean lake (Geneva Lake) and two metal contaminated lakes (Ramsey and Hannah lakes) from Sudbury, ON, in their own lake water or in water from the other lakes. The results showed that fish from the clean lake had a greater olfactory acuity than those from metal contaminated lakes when fish were tested in their own lake water. However, when fish from the clean lake were held for 24h in water from each of the two contaminated lakes their olfactory acuity was diminished. On the other hand, fish from the contaminated lakes held for 24h in clean lake water showed a significant olfactory recovery relative to that measured in their native lake water. These results show that although fish from a clean lake demonstrated impaired olfaction after only 24h in metal-contaminated water, fish from metal contaminated lakes showed a rapid olfactory recovery when exposed to clean water for only hours.

Effects of Continuous Copper Exposure and Calcium on the Olfactory Response of Fathead Minnows

The current gill-based Biotic Ligand Model (gbBLM) is an acute-toxicity model used to predict site-specific safe copper (Cu) concentrations. Recent effort to develop a chronic BLM has focused on the olfactory epithelium. To further this effort, the current study looked at the effect of varying Cu concentration and exposure duration on Cu-induced olfactory dysfunction, and whether calcium (Ca) protected against Cu-induced impairment as it does at the gill. Fathead minnows (Pimephales promelas) were treated with five Cu concentrations for varying exposure durations in hard and soft water. A neurophysiological technique, electro-olfactography (EOG), was employed to determine the level of olfactory dysfunction. At the low, ecologically relevant Cu concentrations tested there was significant inhibition of EOG function; however, over time there was at least a partial recovery of olfactory function, despite the continuous Cu exposure. Calcium did not appear to protect against Cu-induced olfactory dysfunction; and even alone, Ca appeared to interfere with the olfactory response to the amino acid L-arginine. Safe copper concentrations as predicted by the gbBLM, chemosensory-based BLMs, the USEPA BLM, and hardness-adjustment equations based on the exposure waters were not entirely protective against olfactory dysfunction.

Kin recognition and cannibalistic behaviours by adult male fathead minnows (Pimephales promelas)

Parental care is an energetically demanding activity that ensures genes are efficiently passed from one generation to the next. According to evolutionary theory, the greatest energetic investment should be directed towards offspring that are most closely related to the parent. Male fathead minnows, Pimephales promelas, provide this parental investment to developing embryos but not newly hatched larvae. Therefore, selection should favour recognition of embryonic kin to ensure energetic expenditure is optimally invested. In this study, adult male fathead minnows were tested using behavioural assays, with egg cannibalism as an endpoint, to determine whether adult males could discriminate between related and unrelated embryos. Egg cannibalism was highest when adult male fathead minnows were presented with unrelated eggs and lowest when presented with eggs fertilized by the test subject (related eggs). The degree of cannibalism was also a function of breeding status. Unrelated males in breeding condition showed an intermediate response between the low cannibalism demonstrated by related males and the high cannibalism demonstrated by unrelated males in a nonbreeding condition. These results suggest that although male fathead minnows can discriminate between unrelated and related embryos, at least some component of parental investment is a simple function of breeding status.

Biological effects and toxicity of diluted bitumen and its constituents in freshwater systems

Approximately 50 billion cubic meters of bitumen resides within the oil sands region of Alberta, Canada. To facilitate the transport of bitumen from where it is extracted to where it is processed, the bitumen is diluted with natural gas condensate (‘dilbit’), synthetic crude from hydrocracking bitumen (‘synbit’), or a mixture of both (‘dilsynbit’). A primary consideration for the effects of diluted bitumen products on freshwater organisms and ecosystems is whether it will float on the water surface or sink and interact with the stream or lake sediments. Evidence from a spill near Kalamazoo, MI, in 2010 and laboratory testing demonstrate that the nature of the spill and weathering of the dilbit, synbit or dilsynbit prior to and during contact with water will dictate whether the product floats or sinks. Subsequent toxicological data on the effects of dilbit and other diluted bitumen products on freshwater organisms and ecosystems are scarce. However, the current literature indicates that dilbit or bitumen can have significant effects on a wide variety of toxicological endpoints. This review synthesizes the currently available literature concerning the fate and effects of dilbit and synbit spilled into freshwater, and the effects of bitumen and bitumen products on aquatic organisms and ecosystems. Dilbit is likely to provide ecological impacts that are similar to and extend from those that follow from exposure to lighter crude oil, but the prospect of bitumen settling after binding to suspended sediments elevates the risk for benthic impacts in streams and lakes. Copyright