The present study aimed to evaluate biochemical and cellular responses of the freshwater mussel, Hyriopsis bialata, to the herbicide atrazine (ATZ). The mussels were exposed to environmentally-relevant concentrations of ATZ (0, 0.02 and 0.2 mg/L) and a high concentration (2 mg/L) for 0, 7, 14, 21 and 28 days. Tissues comprising male and female gonads, digestive glands and gills were collected and assessed for ethoxyresorufin-O-deethylase (EROD) activity, glutathione S-transferase (GST) activity, multixenobiotic resistance mechanism (MXR), histopathological responses, DNA fragmentation and bioaccumulation of ATZ and its transformation derivatives, desethylatrazine (DEA) and desisopropylatrazine (DIA). Additionally, circulating estradiol levels were determined. It appeared that ATZ did not cause significant changes in activities of EROD, GST and MXR. There were no apparent ATZ-mediated histopathological effects in the tissues, with the exception of the male gonads exhibiting aberrant aggregation of germ cells in the ATZ-treated mussels. Contrarily, ATZ caused significant DNA fragmentation in all tissues of the treated animals in dose- and time-dependent manners. In general, the circulating estradiol levels were higher in the females than in the males. However, ATZ-treated animals did not show significant alterations in the hormonal levels, as compared with those of the untreated animals. Herein, we showed for the first time differentially spatiotemporal distribution patterns of bioaccumulation of ATZ, DEA and DIA, with ATZ and DEA detectable in the gonads of both sexes, DEA and DIA in the digestive glands and only DEA in the gills. The differential distribution patterns of bioaccumulation of ATZ and its derivatives among the tissues point to different pathways and tissue capacity in transforming ATZ into its transformation products. Taken together, the freshwater mussel H. bialata was resistant to ATZ likely due to their effective detoxification. However, using DNA damage as a potential biomarker, H. bialata is a promising candidate for biomonitoring aquatic toxicity.

The environmental neurotoxin β-methylamino-L-alanine (BMAA) may represent a risk for human health in case of chronic exposure or after short-term exposure during embryo development. BMAA accumulates in freshwater and marine organisms consumed by humans. It is produced by marine and freshwater phytoplankton species, but the range of producers remains unknown. Therefore, analysing the phytoplankton composition is not sufficient to inform about the risk of freshwater contamination by BMAA. Filter-feeders mussels have accumulation capacities and therefore appear to be relevant to monitor various pollutants in aquatic ecosystems. We investigated the suitability of the freshwater mussels Dreissena polymorpha and Anodonta anatina for monitoring BMAA in water. Both species were exposed to 1, 10, and 50 μg of dissolved BMAA/L daily for 21 days, followed by 42 days of depuration in clean water. On days 0, 1, 7, 14, and 21 of exposure and 1, 7, 14, 21 and 42 of depuration, whole D. polymorpha and digestive glands of A. anatina were sampled, and the total BMAA concentration was measured. D. polymorpha accumulated BMAA earlier (from day 1 at all concentrations) and at higher tissue concentrations than A. anatina, which accumulated BMAA from day 14 when exposed to 10 μg BMAA/L and from day 7 when exposed to 50 μg BMAA/L. As BMAA accumulation by D. polymorpha was time and concentration-dependent, with a significant elimination during the depuration period, this species may be able to reflect the levels and dynamics of water contamination by dissolved BMAA. The species A. anatina could be used for monitoring water concentrations above 10 μg BMAA/L.

In aquatic environments, organisms such as freshwater mussels are likely exposed to complex contaminant mixtures related to industrial, agricultural, and urban activities. With growing interest in understanding the risk that chemical mixtures pose to mussels, this investigation focused on the effects of various waterborne contaminants (ammonia, chloride, copper, and potassium) and selected binary mixtures of these chemicals following a fixed-ratio design to Villosa iris glochidia and juvenile Lampsilis fasciola. In individual exposures, 48-h EC50 values were determined for V. iris glochidia exposed to ammonia chloride (7.4 [95% confidence interval (CI) 6.6–8.2] mg N/L), ammonia sulfate (8.4 [7.6–9.1] mg N/L), copper sulfate (14.2 [12.9–15.4] μg Cu2+/L), potassium chloride (12.8 [11.9–13.7] mg K+/L), potassium sulfate (10.1 [8.9–11.2] mg K+/L), and sodium chloride (480.5 [435.5–525.5] mg Cl−/L). The 7-d LC50 values for juvenile L. fasciola were determined for potassium sulfate (45.0 [18.8–71.2] mg K+/L), and sodium chloride (1738.2 [1418.6–2057.8] mg Cl−/L). In Ontario these waterborne contaminants have been reported to co-occur, with concentrations exceeding the EC10 for both life stages at some locations. Data from binary mixture exposures for V. iris glochidia (chloride-ammonia, chloride-copper, and copper-ammonia) and juvenile L. fasciola (chloride-potassium) were analyzed using a regression-based, dose-response mixture analysis modeling framework. Results from the mixture analysis were used to determine if an additive model for mixture toxicity [concentration addition (CA) or independent action (IA)] best described the toxicity of each mixture and if deviation towards dose-ratio (DR) or dose-level (DL) synergism/antagonism (S/A) occurred. For all glochidia binary mixture exposures, CA was the best fit model with DL deviation reported for the chloride-copper mixture and DR deviation reported for the copper-ammonia mixture. Using the model deviation ratio (MDR), the observed toxicity in all three glochidia mixture exposures were adequately described by both CA (mean = 0.71) and IA (mean = 0.97) whereas the juvenile mixture exposure was only adequately described by CA (mean = 0.64; IA mean = 0.05).

Due to the increasing presence of platinum (Pt) in the environment, the caveat arises to identify its toxic potential in species at risk of being exposed – especially those found in aquatic environments where pollutants tend to accumulate. Comprehensive characterisation of possible adverse effects following exposure of aquatic organisms to Pt remains elusive. To address this, Zebra mussels (Dreissena polymorpha) were exposed to a range of Pt(IV) concentrations (0.1, 1, 10, 100 and 1000 μg/L) for one and four days, respectively, after which bioaccumulation was quantified and compared to alterations in biomarker profiles relevant to metal toxicity i.e. glutathione-S-transferase (GST) and catalase (CAT) activity, lipid peroxidation and metallothionein (MT) induction. Despite pre-conditioning of the tanks, Pt recovery in the exposure media was found to be 36% (0.1 μg/L), 42% (1 μg/L), 47% (10 μg/L), 68% (100 μg/L) and 111% (1000 μg/L) due to biological and non-biological processes. Pt concentrations in dried mussel soft tissue increased with exposure concentrations and were 20–153 times higher compared to quantified Pt concentrations in the exposure media. CAT activity was significantly increased in the tissue of mussels exposed to 0.1–100 μg/L Pt after Day 1 while the lowest effect concentration (LOC) for this response on both Day 1 and Day 4 was 0.1 μg/L. The effect on the GST activity was less pronounced but demonstrated a similar trend. However, enhanced lipid peroxidation was measured in the tissue of mussels exposed to ≥0.1 μg/L on Day 4. Bioaccumulation of Pt was also associated with a concentration-dependent increase in Pt-MT. Although these effects occurred at Pt levels higher than those present in the environment, it indicates that Pt has the ability to cause aberrancies in metal-associated biomarker profiles.

Freshwater mussels are one of the most threatened taxonomic groups in the world, and many species are on the brink of local or global extinction. Human activities have altered mussel living conditions in a plethora of ways. One of the most destructive human-induced impacts on running waters is the catastrophic spill of harmful substances, which results in massive die-offs. Even though Finland is regarded as the world’s top country in terms of environmental regulation quality, riverine systems are not safe. In 2014, River Kokemäenjoki in western Finland experienced the worst NiSO4 spill in the country’s history, visibly affecting the mussel community – including protected Unio crassus – along the river. Because freshwater mussel toxicology is grossly understudied (particularly in Europe), any pollution –linked die-offs offer valuable opportunities to study the issue in natural environment. Here, we report the mussel investigations from 2014 and a follow-up study conducted in 2017 in order to assess the variation in species sensitivity on nickel pollution. In total, 104 sites were sampled, and over 20 000 mussels were identified and counted. Our results indicate that the most impacted species (i.e. that which experienced the highest spill-induced mortality) was Anodonta anatina (62%), followed by Unio pictorum (32%), U. crassus (24%) and Unio tumidus (9%). The underlying reason for the sensitivity of A. anatina is not resolved, hence more research is urgently needed. The low mortality among most of the species in 2017 highlights the temporal nature of the pollution impact and the recovery potential of the mussel community. However, the case is more complex with U. crassus population, which may be experiencing delayed impacts of the spill. Because nickel is one of the most commonly produced industrial metals in the world (hence the pollution incident risk is high) and River Kokemäenjoki hosts mussel community typical for European rivers, our results may benefit many researchers and stakeholders dealing with riverine environments.

Substituted phenylamines (SPAs) are incorporated into a variety of consumer products (e.g., polymers, lubricants) in order to increase the lifespan of the products by acting as a primary antioxidant. Based on their physicochemical properties, if SPAs were to enter the aquatic environment, they would likely partition into sediment. No studies to date have investigated the effect of sediment-associated SPAs on aquatic organisms. The current study examined the effect of four SPAs (diphenylamine (DPA); N-phenyl-1-napthylamine (PNA); N-(1,3-dimethylbutyl)-N’-phenyl-1,4-phenylenediamine (DPPDA); 4,4’-methylene-bis[N-sec-butylaniline] (MBA)) on three different life stages of the freshwater mussel, Lampsilis siliquoidea. The viability of larvae (glochidia) of L. siliquoidea and Lampsilis fasciola was assessed after 48 h of exposure to SPAs in water. The 48-h EC50s for glochidia viability of L. siliquoidea were 5951, 606, 439, and 258 μg/L for DPA, PNA, DPPDA, and MBA, respectively, and 7946, 591, 137, and 47 μg/L, respectively, for L. fasciola. Juvenile (7–15 months) and adult L. siliquoidea were exposed to sediment-associated SPAs for 28 d. LC50s for juvenile mussels were 18, 55, 62, and 109 μg/g dry weight (dw) of sediment for DPA, PNA, DPPDA, and MBA, respectively. Adult mussels were exposed to sub-lethal concentrations of sediment-associated SPAs in order to investigate reactive oxygen species (ROS), lipid peroxidation and total glutathione in the gill, gonad, and digestive gland tissue, and viability and DNA damage in hemocytes. No significant concentration-dependent trend in any of these biochemical and cellular endpoints relative to the concentration of sediment-associated SPAs was observed in any tissues. Investigations into the concentration of SPAs in the aquatic environment are required before a conclusion can be made on whether these compounds pose a hazard to the different life stages of freshwater mussels.

Freshwater mussels are among the most endangered groups of fauna anywhere in world. The indiscriminate use of antibiotics has led to the emergence of resistant strains. These antibiotic-resistant bacteria play a key role in increasing the risk allied with the use of surface water and in spread of resistance genes. Two endangered freshwater mussel species, Margaritifera margaritifera and Potomida littoralis, were sampled at 4 sampling sites along a 50 km stretch of River Tua. Water samples were taken at same sites. Of the total of 135 isolates, 64.44% (39.26% from water and 25.19% from mussels) were coliform bacteria. Site T1, with the lowest concentration of coliform bacteria, and site T2 were the only ones where M. margaritifera was found. No E. coli isolates were found in this species and the pattern between water and mussels was similar. P. littoralis, which was present at T3/T4 sites, is the one that faces the highest concentration of bacterial toxins, which are found in treated wastewater effluents and around population centers. Sites T3/T4 have the isolates (water and mussels) with the highest resistance pattern, mainly to β-lactams. Water and P. littoralis isolates (T3/T4) showed resistance to penicillins and their combination with clavulanic acid, and to cephalosporins, precisely to a fourth generation of cephalosporin antibiotics. The analysis provides important information on the risk to water systems, as well as the need to investigate possible management measures. It is suggested that future studies on the health status of freshwater bivalves should incorporate measures to indicate bacteriological water quality.

The toxicity of oil sands process-affected water (OSPW) has been primarily attributed to polar organic constituents, including naphthenic acid fraction components (NAFCs). Our objective was to assess the toxicity of NAFCs derived from fresh and aged OSPW, as well as commercial naphthenic acid (NA) mixtures. Exposures were conducted with three aquatic species: Hyalella azteca (freshwater amphipod), Vibrio fischeri (marine bacterium, Microtox® assay), and Lampsilis cardium (freshwater mussel larvae (glochidia)). Commercial NAs were more toxic than NAFCs, with differences of up to 30-, 4-, and 120-fold for H. azteca, V. fischeri, and L. cardium, respectively, demonstrating that commercial NAs are not reliable surrogates for assessing the toxicity of NAFCs. Differences in toxicity between species were striking for both commercial NAs and NAFCs. Overall, V. fischeri was the least sensitive and H. azteca was the most sensitive organism. Responses of V. fischeri and H. azteca to NAFC exposures were consistent (< 2-fold difference) regardless of source and age of OSPW; however, effects on L. cardium ranged 17-fold between NAFCs. NAFCs derived from fresh OSPW sources were similarly or less toxic to those from aged OSPW. Our results support the need to better characterize the complex mixtures associated with bitumen-influenced waters, both chemically and toxicologically.

In temperate urbanized areas where road salting is used for winter road maintenance, the level of chloride in surface waters has been increasing. While a number of studies have shown that the early-life stages of freshwater mussels are particularly sensitive to salt; few studies have examined the toxicity of salt-impacted winter road runoff to the early-life stages of freshwater mussels to confirm that chloride is the driver of toxicity in this mixture. This study examines the acute toxicity of field-collected winter road runoff to the glochidia of wavy-rayed lampmussels (Lampsilis fasciola) (48 h exposure) and newly released juvenile fatmucket mussels (Lampsilis siliquoidea) (<1 week old; 96 h exposure) under different water hardness. The chronic toxicity (28 d) to older juvenile L. siliquoidea (7–12 months old) was also investigated. The 48-h EC50 and 96-h LC50 for L. fasciola glochidia and L. siliquoidea juveniles exposed to different dilutions of road run-off created with moderately hard synthetic water (∼80 mg CaCO3/L) were 1177 (95% confidence interval (CI): 1011–1344 mg Cl−/L) and 2276 mg Cl−/L (95% CI: 1698–2854 mg Cl−/L), respectively. These effect concentrations correspond with the toxicity of chloride reported in other studies, indicating that chloride is likely the driver of toxicity in salt-impacted road-runoff, with other contaminants (e.g., metals, polycyclic aromatic hydrocarbons) playing a de minimis role. Toxicity data from the current study and literature and concentrations of chloride in the surface waters of Ontario were used to conduct a probabilistic risk assessment of chloride to early-life stage freshwater mussels. The assessment indicated that chronic exposure to elevated chloride levels could pose a risk to freshwater mussels; further investigation is warranted to ensure that the most sensitive organisms are protected.

Cyphenothrin, one of the synthetic pyrethroids, developed as an alternative to organophosphorus and carbamate pesticides. It is used in veterinary medicine and household application against insects. Due to the contamination of the aquatic ecosystems, non-target aquatic organisms are affected. The current study aimed to evaluate the cyphenothrin effects on in vitro and in vivo models of freshwater mussels Unio delicatus Lea, 1863. While antioxidant enzyme (glutathione) was measured in both models, the total hemocyte counts were only detected in vivo models after exposure to cyphenothrin (1 and 10 μg/L) for 24-h and 48-h exposure times. A decrease in total hemocyte count occurred depending on the dose and duration (p < 0.001). In both in vitro and in vivo models of gill and digestive gland tissues, higher glutathione levels were obtained at a dose of 10 μg/L compared to the control groups in both exposure times (p < 0.001). The results of the study suggest that the antioxidant parameters could represent biomarkers to evaluate the effects of pollutants on in vitro and in vivo models of freshwater mussels.