Interaction of the Olfactory System of Rainbow Trout (Oncorhynchus mykiss) with Diltiazem.

Diltiazem is ubiquitously prescribed and has been reported in many effluents and freshwater bodies. Being a calcium channel blocker, diltiazem could disrupt the function of the sensory and central nervous systems. In the present study, using electro-olfactography (EOG), we investigated the interaction of diltiazem with the olfactory sensory neurons (OSNs) of rainbow trout by looking into the detection threshold and effects of immediate (~5 min) and acute (24 h) exposure to diltiazem at 6.6, 66, and 660 µg/L. We also studied the accumulation of the drug in fish plasma and whole body. Brief exposure to diltiazem impaired the OSN response to a chemosensory stimulus in a concentration-dependent manner at 6.6 µg/L and higher, whereas OSNs exposed for 24 h only displayed an impairment at 660 µg/L. Chemical analysis showed that the accumulation of diltiazem in fish plasma and body correlated with the EOG response because it was 10 times higher in the group that displayed a significant impairment (660 µg/L) compared to the other 2 groups (6.6, 66 µg/L). This correlation suggests that the impact of diltiazem on OSNs might partially be through the accumulated molecules in the fish bloodstream. Fish did not detect diltiazem as a sensory stimulus even at concentrations as high as 660 µg/L; thus, fish could potentially swim toward or fail to escape harmful concentrations of diltiazem. Environ Toxicol Chem 2022;41:554-550. © 2020 SETAC.

The effects of diltiazem on growth, reproduction, energy reserves, and calcium-dependent physiology in Daphnia magna.

With the growth of both the pharmaceutical industry and the human population and longevity, more drugs are used and processed each day. Inevitably, these pharmaceuticals enter wastewater through human excretion and improper disposal of leftovers. One such medication, diltiazem, a calcium channel blocker, is of importance due to its widespread consumption, and prevalence in aquatic environments. To study the sub-lethal effects of diltiazem on aquatic animals, we investigated its impacts no feeding behaviour, heart rate, respiration, growth, and reproduction of a bioindicator species, Daphnia magna. When exposed to environmentally relevant concentrations, D. magna increased their heart rate by 12% and oxygen consumption by 48%. However, exposure did not have any effects on thoracic limb movement frequency or peristalsis (i.e. feeding behaviour). Individuals exposed to diltiazem for a longer duration (16 days) showed a 44% decrease in lipid reserves and produced between 17 and 28% fewer neonates which were 10-12% larger. Our study demonstrated that exposure to diltiazem creates an energy imbalance in D. magna which could, in the long run, influence their populations.

Oil sands process-affected water impairs the olfactory system of rainbow trout (Oncorhynchus mykiss).

Oil sands process-affected water (OSPW), a byproduct of the extraction of bitumen in the surface mining of oil sands, is currently stored in massive on-site tailings ponds. Determining the potential effects of OSPW on aquatic ecosystems is of main concern to oil sands companies and legislators concerned about the reclamation of mining sites. In the present study, the interaction of OSPW with the chemosensory system of rainbow trout was studied. Using an electro-olfactography (EOG) technique, a 24 h inhibition curve was established and concentrations that inhibit the olfactory system by 20% and 80% (IC20 and IC80) were estimated at 3% and 22% OSPW, respectively. To study the interaction of exposure time and concentration along with the mechanism of the toxic effects, rainbow trout were exposed to 3% and 22% OSPW for 2, 24, and 96 h. An EOG investigation of olfactory sensitivity demonstrated a positive interaction between exposure time and concentration of OSPW concentration, because an increase in either or both elevated the inhibitory effect. To investigate whether or not structural damage of the olfactory epithelium could account for the observed inhibitory effects of OSPW on fish olfaction, the ultrastructure of the olfactory epithelium of exposed fish was investigated using scanning electron microscopy (SEM) and light microscopy (LM). The SEM micrographs showed no changes in the structure of the olfactory epithelium. The light micrographs revealed an increase in the number of mucous cells in 22% OSPW. The results of the present study demonstrated that exposure to OSPW impairs the olfactory system of rainbow trout and its effects increase gradually with increasing exposure time. The present study demonstrated that structural epithelial damage did not contribute to the inhibitory effects of OSPW on the olfactory system.

Effects of Seasonal Changes on the Toxic Impacts of Oil Sands Process-Affected Water on Daphnia magna.

Oil sands process-affected water (OSPW), which can be potentially toxic to aquatic biota, is a major by-product of bitumen mining in northern Alberta. The effects of environmental factors on the toxicity of OSPW are understudied. In the present study, the impacts of seasonal changes in water quality on the toxic effects of OSPW (1 and 10%) on Daphnia magna was examined. Animals were chronically exposed to OSPW under conditions that represented water quality of a cold or warm seasonal condition. At each seasonal scenario survival, growth (length and mass) and reproduction of exposed D. magna were investigated. Survival and length of D. magna were only affected by OSPW in the cold-season treatment. Exposure to OSPW reduced the mass of D. magna in both cold and warm season scenarios. Daphnia magna in the cold-season treatment did not reproduce or produce eggs during the course of the experiment. The results of the present study suggest that seasonal changes in water quality may alter the toxicity of OSPW on D. magna.

Effects of gemfibrozil on the growth, reproduction, and energy stores of Daphnia magna in the presence of varying food concentrations.

Gemfibrozil, a common lipid regulator, enters aquatic environments through treated municipal wastewater effluent that fails to remove it completely from effluent streams. When exposed to gemfibrozil concentrations of 50, 500, 5,000, and 50,000 ng L-1, Daphnia magna showed increased lipid reserves by 14-21% (significant at 500 ng L-1), increased length by 9-13% (significant at 50 ng L-1), increased mass by 6-13% (significant at 50 ng L-1) and increased neonate production by 57-74% (significant at 50 ng L-1). Gemfibrozil-exposed Daphnia held under conditions where food availability was low, grew and reproduced as well as those in the control. Taken together, these results suggest that gemfibrozil exposure within environmentally relevant concentration ranges is not toxic to Daphnia magna but has the potential to be beneficial to the species under these conditions.

Investigating the chronic effects of oil sands process-affected water on growth and fitness of Daphnia magna Straus 1820.

The increasing amount of stored oil sands process-affected water (OSPW), a primary by-product of oil sands mining, is an environmental concern. In the present study, we investigated the chronic effects of OSPW on growth, reproduction, and macronutrient content in Daphnia magna. To do so, we exposed D. magna to 1 and 10% OSPW (a mixture of three OSPW samples provided by major oil sands mining operators in northern Alberta) for ten days. We measured the number of the neonates produced daily in each group throughout the exposure. At the end of the exposure, we measured the mass and length of the exposed daphniids and neonates. We also measured the carbohydrate, lipid, and protein content of exposed daphniids. In the 10% OSPW group, we observed a significant reduction in all of the measured endpoints except for body length and carbohydrate and protein content of exposed daphniids. In the 1% OSPW group, on the other hand, we found a reduction only in lipid content of exposed daphniids as compared to the control group. The results of the present study demonstrated that chronic exposure to 10% OSPW affects growth and fitness of D. magna, probably due to a reduction in energy intake that causes daphniids to deplete their energy reserves.

Oil sands process-affected water impairs feeding by Daphnia magna.

Growth in extraction of bitumen from oil sands has raised concerns about influences of this industry on surrounding environments. Water clearance rate (a surrogate of feeding rate by Daphnia magna) in water containing D. magna exposed to oil sands process-affected water (OSPW) and its principal components, dissolved component (DC) and suspended particulate matter (SPM), was reduced to 72, 29, and 59% of controls, respectively. This study also examined several possible mechanisms for the observed changes algal cell density (i.e., feeding rate). There was no change in the digestive enzymes trypsin or amylase when D. magna were exposed to DC or SPM; however, exposure to total OSPW reduced trypsin activity. Mandible rolling or post-abdominal rejections, which are indicators of feeding and palatability of food, were not affected by any exposures to OSPW. Beating of thoracic limbs, which provides water flow toward the feeding groove, was reduced by exposure to SPM or total OSPW. Peristaltic activity was reduced by exposure to DC, which then might result in reduced digestion time in D. magna exposed to DC, SPM or whole OSPW. All treatments caused an increase in numbers of intact algae cells in the hindgut and excreted material. These results suggest that both DC and SPM affect feeding of D. magna by impairing actions of the digestive system, but most probably not by reducing rates of ingestion.

A novel apparatus for evaluating contaminant effects on feeding activity and heart rate in Daphnia spp.

Cladoceran are animals of significant importance to freshwater bodies such that changes in their populations may result in drastic shifts in the food web. Numerous studies have investigated the effects of toxicants, and the effects of chemical and physical habitat changes to these animals. Most of these studies investigated more general endpoints such as mortality, reproduction, growth, and food consumption over time, and less frequently examined molecular endpoints such as enzyme activity or gene expression. However, behavioral and physiological endpoints that link the organism and molecular level endpoints are scarce. In this study, we designed an apparatus that allows for the simultaneous investigation of three essential behavioral and physiological endpoints in Daphnia, including ventilation, food uptake rate, and heart rate. Using our apparatus, we studied the effect of cadmium (Cd), suspended particles, and food on the beating rate of thoracic limbs and the frequency of mandible rolling in Daphnia magna. We also studied the effect of temperature on the heart and thoracic limb beat frequency. The results show that both Cd and suspended particles reduce the activity of mandibles and thoracic limbs. Thoracic limb movements and heart rate increased gradually with temperature. Our toxicity tests show that changes in feeding, ventilation, and heart rate are easily detected using this method.

Rapid changes in water hardness and alkalinity: Calcite formation is lethal to Daphnia magna.

There is growing concern that freshwater ecosystems may be negatively affected by ever-increasing anthropogenic inputs of extremely hard, highly alkaline effluent containing large quantities of Ca(2+), Mg(2+), CO3(2-), and HCO3(-) ions. In this study, the toxicity of rapid and extreme shifts in water hardness (38-600mg/L as CaCO3) and alkalinity (30-420mg/L as CaCO3) to Daphnia magna was tested, both independently and in combination. Within these ranges, where no precipitation event occurred, shifts in water hardness and/or alkalinity were not toxic to D. magna. In contrast, 98-100% of D. magna died within 96h after exposure to 600mg/L as CaCO3 water hardness and 420mg/L as CaCO3 alkalinity (LT50 of 60h with a 95% CI of 54.2-66.0h). In this treatment, a CaCO3 (calcite) precipitate formed in the water column which was ingested by and thoroughly coated the D. magna. Calcite collected from a mining impacted stream contained embedded organisms, suggesting field streams may also experience similar conditions and possibly increased mortality as observed in the lab tests. Although further investigation is required to determine the exact fate of aquatic organisms exposed to rapid calcite precipitation in the field, we caution that negative effects may occur more quickly or at lower concentrations of water hardness and alkalinity in which we observed effects in D. magna, because some species, such as aquatic insects, are more sensitive than cladocerans to changes in ionic strength. Our results provide evidence that both calcite precipitation and the major ion balance of waters should be managed in industrially affected ecosystems and we support the development of a hardness+alkalinity guideline for the protection of aquatic life.