Optimizing Sex Ratios of Hyalella azteca to Reduce Variability in Reproduction and Improve Reproductive Toxicity Test Methods.

Standard toxicological test methods with Hyalella azteca incorporate both lethal and sublethal (growth, reproduction) endpoints, although lethal endpoints are often favored in routine testing. However, sublethal endpoints are important to consider because they are ecologically relevant and are often more sensitive than lethality. It is difficult to achieve robust data for reproduction in H. azteca because high biological variability is associated with reproductive yield, likely due to complex interactions in mate selection, which can add to the variability in brood sizes that females produce. In addition, effects on reproduction often co-occur with effects on growth, making it difficult to differentiate effects on growth from those on reproduction in standard tests initiated with juveniles. The present study characterized the reproductive capacity of H. azteca by investigating the role of sex ratios in reproductive yield. Experiments were initiated in the absence of toxicants with sexually mature (6-8-week-old) individuals that were placed in different female-to-male ratios (1:1, 2:3, 3:2, and 7:3). Reproduction was monitored during weekly static renewals for 7 weeks. The results indicated that a higher female:male ratio (seven females to three males) improved reproductive success and lowered biological variability in reproduction. In addition, the body lengths and weights of newly mature amphipods were used to create a size distribution to aid in the identification of small female amphipods. Based on the results of our study, a novel reproductive toxicity test method is proposed that begins with sexually mature amphipods in a ratio of seven females to three males to minimize reproductive variability in amphipods and to enable a more effective assessment of contaminants using reproduction as a highly sensitive endpoint. However, evaluation of this novel method with toxicants is needed to demonstrate that results are comparable to standard methods. Environ Toxicol Chem 2024;43:712-722. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Toxicity of 6PPD-quinone to four freshwater invertebrate species.

The antioxidant N-(1,3-Dimethylbutyl)-N'-phenyl-p- phenylenediamine (6PPD) is used to protect the rubber in tires from oxidation, which extends the life of the tire. When oxidized, 6PPD is transformed into 6PPD-quinone (6PPDQ). 6PPDQ, along with other tire ingredients, can enter aquatic ecosystems through the transport of tire wear particles in runoff during a precipitation event. The mass mortality of coho salmon following precipitation events in urban areas lead to the discovery that 6PPDQ is the likely cause due to coho salmon's relatively high sensitivity to 6PPDQ. The assessment of 6PPDQ toxicity to other aquatic species has expanded, but it has focused on fish. This study investigated the toxicity of 6PPDQ to four freshwater invertebrate species, larval burrowing mayfly (Hexagenia spp.), juvenile cladoceran (Daphnia magna), file ramshorn snail embryo (Planorbella pilsbryi), and adult washboard mussel (Megalonaias nervosa). For all four species, the highest concentration of 6PPDQ tested did not result in significant mortality. This translated into the determination of the highest concentration that did not cause significant mortality (NOEC) for Hexagenia spp., D. magna, P. pilsbryi, and M. nervosa of 232.0, 42.0, 11.7, and 17.9 µg/L, respectively. The data from this study indicate that freshwater invertebrates are not as sensitive to 6PPDQ as some salmonid species (e.g., coho salmon Oncorhynchus kisutch). This study also analyzed 6PPDQ in road runoff from around the city of Guelph in Ontario, Canada. 6PPQ was detected in all samples but the concentration was two orders of magnitude lower than the NOECs for the four tested species of freshwater invertebrate.

Discerning conformational dynamics and binding kinetics of GPCRs by 19F NMR.

19F NMR provides a way of monitoring conformational dynamics of G-protein coupled receptors (GPCRs) from the perspective of an ensemble. While X-ray crystallography provides exquisitely resolved high-resolution structures of specific states, it generally does not recapitulate the true ensemble of functional states. Fluorine (19F) NMR provides a highly sensitive spectroscopic window into the conformational ensemble, generally permitting the direct quantification of resolvable states. Moreover, straightforward T1- and T2-based relaxation experiments allow for the study of fluctuations within a given state and exchange between states, on timescales spanning nanoseconds to seconds. Conveniently, most biological systems are free of fluorine. Thus, via fluorinated amino acid analogues or thiol-reactive fluorinated tags, F or CF3 reporters can be site specifically incorporated into proteins of interest. In this review, fluorine labeling protocols and 19F NMR experiments will be presented, from the perspective of small molecule NMR (i.e. drug or small molecule interactions with receptors) or macromolecular NMR (i.e. conformational dynamics of receptors and receptor-G-protein complexes).

Development of an Embryo Toxicity Test to Assess the Comparative Toxicity of Metal Exposure on Different Life Stages of Freshwater Gastropods.

Early life stages are commonly thought to be highly sensitive to environmental contaminants and may offer insight into the future health of a population. Despite the importance of studying early life stages, very few standard protocols for benthic invertebrates commonly used in ecotoxicological assessments measure developmental endpoints. The goal of the present study was to develop and optimize a robust standard protocol for studying embryonic endpoints in freshwater gastropods. The developed method was then used to characterize the sensitivity of four embryonic endpoints (viability, hatching, deformities, and biomass production), in conjunction with juvenile and adult mortality, for the snail Planorbella pilsbryi exposed to three metals (copper [Cu], cadmium [Cd], and nickel [Ni]). Biomass production was typically the most sensitive endpoint but was relatively variable, while embryo hatching was slightly less sensitive but highly consistent for all three metals. However, no single embryonic endpoint was consistently the most sensitive, which demonstrates the importance of assessing a broad range of endpoints and life stages in ecotoxicological risk assessment. Interestingly, the embryonic life stage of P. pilsbryi was considerably less sensitive to Cu exposure compared with juvenile and adult mortality. However, for Cd exposure, embryonic endpoints were the most sensitive, and for Ni exposure, embryonic endpoints were similar in sensitivity to juvenile and adult mortality. The present study has valuable applications in conducting developmental toxicity research with organisms lacking standardized testing protocol as well as future applications in multigenerational and in silico toxicity research. Environ Toxicol Chem 2023;42:1791-1805. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Potential risk to aquatic biota from aerial application of firefighting water additives.

The frequency and severity of forest fires is increasing due to climate change. Consequently, there will be an increased use of forest firefighting additives, which increase the ability of water to extinguish fires and prevent reignition. Increased use will potentially result in increased exposure to aquatic ecosystems within forests. This study examined the toxicity of nine firefighting water additives that are currently on the market to three species of freshwater invertebrates that occupy different niches within freshwater ecosystems. The toxicity of the water additives varied up to three orders of magnitude. Pelagic and epibenthic invertebrates are affected at lower rates of application than endobenthic invertebrates. A field relevant application rate of three of the nine water additives tested represent a hazard to freshwater ecosystems under varies exposure scenarios represented by the depth of a theoretical water body (15-200 cm). This study highlights the importance of application buffers around water bodies and the selection of water additives that pose the lowest hazard to freshwater ecosystem, assuming that the efficacy of the additives in extinguishing fires is similar.

Investigation of the potential effects of firefighting water additives on soil invertebrates and terrestrial plants.

The intensity and frequency of forest fires is increasing across the globe due to climate change. Additives are often added to make water more effective at extinguishing fire and preventing re-ignition. This study investigated the toxicity of nine different firefighting water additives to four species of soil invertebrates (Folsomia candida, Porcellio laevis, Porcellio scaber, and Trichorhina tomentosa) and two plant species (Agropyron cristatum and Raphanus sativus). Considerable variation in toxicity was observed among the firefighting products. The toxicity of individual products also varied considerably amongst the tested species. A hazard assessment was conducted by comparing the concentration of firefighting water additive that caused a 50% effect (LC50 or EC50) or a concentration that caused no effect (NOEC) to the concentration recommended by the manufacturer. At a rate of application representative of a forest firefighting scenario, most firefighting water additives tested in this study posed a hazard to F. candida and the three isopod species. The majority of products did not pose a risk to the two plant species included in this study. Consideration of the toxicity of firefighting water additives to terrestrial biota should be considered along with the efficacy of the product to fight fires when deciding which products to use.

Fate of thiamethoxam from treated seeds in mesocosms and response of aquatic invertebrate communities.

Thiamethoxam is a neonicotinoid insecticide widely applied in the Canadian Prairies. It has been detected in surface waters of agro-ecosystems, including wetlands, but the potential effects on non-target invertebrate communities in these wetlands have not been well characterized. In an effort to understand better the fate of thiamethoxam in wetlands and the response of invertebrates (zooplankton and emergent insects), model systems were used to mimic wetland flooding into planted fields. Outdoor mesocosms were treated with a single application of thiamethoxam-treated canola seeds at three treatment levels based on a recommended seeding rate (i.e., 6 kg/ha; 1×, 10×, and 100× seeding rate) and monitored over ten weeks. The mean half-life of thiamethoxam in the water column was 6.2 d. There was no ecologically meaningful impact on zooplankton abundances or community structure among treatments. Statistically significant differences were observed in aquatic insect abundance between control mesocosms and the two greatest thiamethoxam treatments (10× and 100× seeding rate). The observed results indicate exposure to thiamethoxam at environmentally relevant concentrations likely does not represent a significant ecological risk to abundance and community structure of wetland zooplankton and emergent insects.

Bioaccumulation of sediment-associated dinonylnaphthalene sulfonates in the freshwater mussel Lampsilis siliquoidea and oligochaete Tubifex tubifex.

Naphthalene sulfonic acids (NSAs) are used primarily as additives in a wide range of industrial products (e.g., rubber materials, coatings, sealants, fuels, paints). Based on modeled physicochemical properties, NSAs would likely partition into sediments or the tissues of biota in an aquatic system. This study examined the potential for three NSAs, dinonylnaphthalene disulfonic acid (DNDS), barium dinonylnaphthalene sulfonate (BaDNS), and calcium dinonylnaphthalene sulfonate (CaDNS), to accumulate in the tissue of a freshwater mussel (Lampsilis siliquoidea) and oligochaete worm (Tubifex tubifex). The ability of L. siliquoidea to depurate accumulated chemical was also assessed. Mussels were exposed via sand spiked with CaDNS for 25 d, and then transferred to clean water where their ability to depurate the chemical over an additional 28 d was monitored. Worms were exposed to each of the three NSAs via spiked sediment for 28 d. NSA concentrations were measured separately in gill, foot, and remaining soft tissues (viscera) for mussels and in whole body tissue samples of worms. For L. siliquoidea, the largest concentration of CaDNS was measured in the gill tissue; once removed from CaDNS exposure, mussels were able to depurate up to 87% of the CaDNS from their tissues in 28 days. The biota-sediment accumulation factors (28-d BSAFs) for T. tubifex were 2.8-5.2, 0.53-0.76, and 0.83-1.11 for DNDS, BaDNS, and CaDNS, respectively. For mussel gill and viscera, BCFK values were 14.07 and 16.39, respectively. When BAFKs were calculated using the concentration of CaDNS in sand, they were 1.11 and 1.29 for mussel gill and viscera, respectively. These values are much lower than what would be necessary to classify this chemical as bioaccumulative; however, the BSAFs for DNDS in T. tubifex indicated a potential biomagnification concern if this compound were to occur in the aquatic environment.

A probabilistic risk assessment of microplastics in soil ecosystems.

Plastics have a variety of applications due to their versatility, relative cost, and strength-to-weight ratio, and resistance to degradation. As a result, plastic waste can be found in all corners of the Earth. A class of plastic contaminants that have received increasing attention in terms of their potential impact on ecosystems is microplastics (≤5 mm). The greatest attention to date has been on their potential effect in marine ecosystems. However, a growing number of studies are examining their potential impact on soil ecosystems. The data reported in the literature on the environmentally-relevant concentrations of microplastics in soils and the concentration of microplastics that causes an adverse effect in soil biota were used to perform a probabilistic risk assessment of microplastics to soil biota. An environmental exposure distribution was constructed from the concentrations of microplastics reported in soil in the literature. Species sensitivity distributions were constructed using concentration of microplastics in soil that had no adverse effect on soil species (NOEC) or the lowest concentrations that had an adverse effect on soil species (LOEC) reported in the literature. The 95th centile of the environmental exposure distribution (8147 microplastic particles per gram of soil) was greater than 22 and 28% of the species sensitivity distribution constructed using NOECs and LOECs, respectively. The assessment concluded that environmentally relevant concentrations of microplastics reported in the literature could pose a considerable risk to soil biota. It is also important to note that due to the continued production of large quantities of plastic and the persistence of microplastics in the environment, environmentally-relevant concentrations of microplastics in soil are likely to only rise.

The influence of organic carbon on the toxicity of sediment-associated dinonylnaphthalene sulfonic acids to the benthic invertebrates Tubifex tubifex and Hyalella azteca.

Naphthalene sulfonic acids (NSAs) are used extensively in industrial applications as dispersants in dyes, rubbers, and pesticides, and as anti-corrosive agents in coatings, gels, and sealants. This study examined the toxicity of three NSA congeners, barium dinonylnaphthalene sulfonate (BaDNS), calcium dinonylnaphthalene sulfonate (CaDNS), and dinonylnaphthalene disulfonic acid (DNDS), to two benthic species, Tubifex tubifex and Hyalella azteca. Two substrates with different levels of organic carbon (sediment [2%] and sand [0%]) were used in toxicity tests. Juvenile production was the most sensitive endpoint for T. tubifex: the 28-d EC50s were <18.2, 22.2, and 64.0 µg/g dw in sand and 281.3, 361.6, and 218.9 µg/g dw in sediment for BaDNS, CaDNS, and DNDS, respectively. The 28-d LC50s for H. azteca were similar among compounds: 115.3, 82.1, and 49.0 µg/g dry weight (dw) in sand, and 627.3, 757.9, and >188.5 µg/g dw in sediment, for BaDNS, CaDNS, and DNDS, respectively. However, when LC50s were estimated based on concentrations of NSAs measured in overlying water (which can be an important route of exposure for H. azteca), BaDNS and CaDNS were 3-4 orders of magnitude more toxic than DNDS. The NSAs examined were >3-fold more toxic when present in substrates with no organic carbon (e.g., sand) for all H. azteca endpoints where LC/EC50s could be calculated and for sublethal endpoints for T. tubifex. The organic carbon content of the sediment appears to have acted as a sink and reduced NSA toxicity by decreasing bioavailability. Environmental sediment samples were collected from 12 river sites across southern Ontario. The maximum concentration of CaDNS observed in sediment collected from this region was 2.8 µg/g dw in sediment with 2% organic carbon; 100-fold lower than the lowest EC10 in the current study.

High-Frequency Sampling of Small Streams in the Agroecosystems of Southwestern Ontario, Canada, to Characterize Pesticide Exposure and Associated Risk to Aquatic Life.

The temporal dynamics of pesticide concentrations in streams remains poorly characterized in southwestern Ontario, a region of the province where land use is dominated by agriculture. Understanding the magnitude and duration of pulsed exposures to pesticides in these small streams is critical when estimating the risk of pesticides to these aquatic ecosystems. The present study investigated the application of a high-frequency water sampling approach paired with the collection of flow data to characterize the pulsed exposure of pesticides to small streams in southwestern Ontario. Six sites along 2 different streams with different magnitudes of agricultural land use in their upstream catchments were sampled using half-day composite samples from July to October 2018 and from May to September 2019. A total of 1043 samples were collected over the 2 yr, of which 210 were analyzed. Samples for analysis were chosen based on flow, water level, and precipitation data. Liquid and gas chromatography coupled with tandem mass spectrometry was used to measure >500 pesticides in each water sample. A total of 35 different compounds were detected over the 6 sampling sites. For pesticides that were detected in >10% of water samples above the method quantification limit, a deterministic risk assessment using water quality guidelines and a probabilistic risk assessment using species sensitivity distributions were performed. The calculated hazard quotients showed that 2,4-D, atrazine, metolachlor, and metribuzin exceeded a level of concern of 1 at the highest concentrations detected. In all cases, hazard concentrations that would be protective of 95% of species from the species sensitivity distributions were greater than the 95th centile of the environmental exposure distributions, meaning that the risk from the pesticides was low. Environ Toxicol Chem 2020;39:2570-2587. © 2020 SETAC.

Toxicity of dinonylnaphthalene sulfonates to Pimephales promelas and epibenthic invertebrates.

Dinonylnaphthalene sulfonic acids (NSAs) are high production volume chemicals that are used primarily as additives in a wide range of industrial products (i.e., coatings, sealants, fuels, metal-extractants, paints, rubber materials). This study examined the effect of three NSA congeners on freshwater organisms: barium dinonylnaphthalene sulfonate (BaDNS), calcium dinonylnaphthalene sulfonate (CaDNS), and dinonylnaphthalene disulfonic acid (DNDS). Chronic effects were characterized by exposing fertilized fathead minnow eggs to sediment-associated NSAs and measuring various developmental and growth endpoints for 21 d. No effects in hatch success and larval growth were observed when fathead minnow eggs were exposed to CaDNS and DNDS concentrations up to 246 and 798 µg/g dry weight, respectively, in spiked sediment (~2% organic carbon). However, when NSAs were associated with substrate containing no organic carbon (sand), EC50s for fathead minnow hatch success, larval growth, biomass production, and overall survival were 58.3, 18.8, 15.5, and 13.8 µg/L, respectively, for CaDNS. Acute effect characterization was also conducted in water-only exposures for the three NSA congeners using the freshwater amphipod Hyalella azteca, the pulmonate snail Planorbella pilsbryi, and larval freshwater mussels Lampsilis cardium and Lampsilis siliquoidea. The sulfonate salts (BaDNS and CaDNS) were significantly more acutely toxic to all tested invertebrates in the water-only exposures, with LC50s ranging from 0.47 to 12.1 µg/L, compared to DNDS (LC50s ≥ 98.2 µg/L). This is the first study to provide empirical data on the aquatic toxicity of three NSA congeners.

Deterministic risk assessment of firefighting water additives to aquatic organisms.

Past firefighting water additives were found to contain perfluorinated compounds that could persist in the environment resulting in potential adverse effects to biota. Since this revelation, manufacturers have introduced alternative firefighting water additives that are fluorine free, but few studies have investigated the fate and effects in the environment of these new additives. Firefighting water additives could enter aquatic ecosystems through run-off, leaching or direct application. Therefore, there is a need to investigate the potential effect that firefighting water additives could have on aquatic biota. This study investigated the toxicity of six firefighting water additives: Eco-Gel™, Thermo-Gel™, FireAde™, Fire-Brake™, Novacool Foam™, and F-500™ to aquatic biota. The toxicities of firefighting water additives to Lemna minor (duckweed), Daphnia magna (water flea), Hexagenia spp. larvae (mayfly), Lampsilis fasciola (wavy-rayed lampmussel) and Oncorhynchus mykiss (rainbow trout) were investigated through acute and chronic static and semi-static tests to estimate LC50 values for survival and EC50 values for immobility and/or reproduction endpoints. A large variation in toxicities among the firefighting water additives and among the test species was observed. Based on a worst-case exposure scenario of direct application, several firefighting water additives were found to pose a hazard to aquatic organisms. An exposure rate representative of a direct overhead application by a water bomber during a forest fire was used in the hazard assessment. For example, the hazard quotients determined for the D. magna acute toxicity tests ranged from 0.20 for Eco-Gel to 317 for F-500 in the forest pool (15 cm) scenario. This study presents the first deterministic risk assessment of firefighting water additives in aquatic ecosystems.

A review of the effectiveness of vegetated buffers to mitigate pesticide and nutrient transport into surface waters from agricultural areas.

A relatively large number of studies have investigated the effectiveness of vegetated buffer strips at reducing the movement of pesticides and nutrients from agriculture fields. This review outlines the observed influence of different factors (e.g., buffer width, slope, runoff intensity, soil composition, plant community) that can influence the efficacy of vegetated buffers in pesticide and nutrient retention. The reported effectiveness of vegetated buffers reducing the movement of pesticides and nutrients ranged from 10 to 100% and 12-100%, respectively. Buffer width is the factor that is most frequently considered by various jurisdictions when making recommendations on vegetated buffer strip implementation. However, the literature clearly illustrates that there is a great deal of variation in pesticide or nutrient reduction for a given buffer width. This indicates that other factors play an important role in buffer efficacy (e.g., ratio of source area to buffer area, soil composition and structure, runoff intensity, plant community structure) in addition to the width of the vegetative buffer area. These factors need to be considered when making recommendations on vegetated buffer strip construction in agroecosystems. This review has also identified a number of other gaps in the understanding of the effectiveness of vegetated buffers at reducing the movement of pesticides and nutrients from the areas of application.

Sensitivity of larval and juvenile freshwater mussels (unionidae) to ammonia, chloride, copper, potassium, and selected binary chemical mixtures.

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).

The role of vegetated buffers in agriculture and their regulation across Canada and the United States.

A vegetated buffer, barrier, or filter strip is a parcel of land that is designated to separate land used for agriculture from valued aquatic or terrestrial habitats. It exists partly with the intent to diffuse runoff and to impeded sediment, nutrients, pesticides, and other constituents from reaching off-site surface waters. Mandatory buffer implementation is regulated at various levels of government in North America - from the federal to the state and provincial levels, and by some municipalities and counties. To better understand the degree and breadth of oversight, we undertook a comprehensive search and review of vegetative buffer regulations across North America. We determined the width of buffer required, under what habitat or field conditions, for which pesticides, and application type, amongst other attributes. For ground application, margins ranged from 1 m to upwards of greater than 4000 m depending on protection goals, with some being compound specific and others being generally applied to all registered pesticides/compounds. These buffers tended to be used most often to protect surface water, groundwater (e.g. drinking water wells), and nearby sensitive crops, but the required distances are generally not consistent between jurisdictions, regardless of the stated protection goals. We recommend that a thorough science-based review take place, with input from relevant stakeholders, to harmonize vegetated buffer size for effective surface water protection where ecological, climatic, and agricultural conditions are sufficiently similar in North America.

The facultatively parasitic ciliated protozoan, Tetrahymena glochidiophila (Lynn, 2018), causes a reduction in viability of freshwater mussel glochidia.

This study investigated the effect of a previously uncharacterized species of ciliated protozoan, Tetrahymena glochidiophila, on the viability of glochidia from three species of freshwater mussel (Lampsilis siliquoidea, Lampsilis fasciola, and Lampsilis cardium). Over the course of 72 h, the viability of glochidia exposed to T. glochidiophila declined by >60% while the decline in the viability of uninfected glochidia was <10%. The density of T. glochidiophila increased >1000-fold during the experiment in treatments with infected glochidia. Lampsilis cardium glochidia were also either exposed to gill rinsate or gill contents from infected gravid female L. siliquoidea for the purpose of elucidating the location of the greatest density of ciliates within infected mussels. Glochidia exposed to gill contents declined significantly (p < 0.05) more than glochidia exposed to gill rinsate. Finally, a clone of ciliates was isolated from infected glochidia and cultured on bacterized medium. The clonal culture was then used to expose uninfected glochidia for the purpose of further confirming a parasitic relationship between glochidia and T. glochidiophila. The viability of glochidia exposed to T. glochidiophila from the clonal culture declined significantly relative to uninfected glochidia but not to the extent of glochidia exposed to ciliates from the gills of infected L. siliquoidea.

Fate of thiamethoxam in mesocosms and response of the zooplankton community.

Thiamethoxam is a neonicotinoid insecticide that can reach wetlands in agro-ecosystems through runoff. The fate and effects of thiamethoxam on non-target organisms in shallow wetland ecosystems have not been well characterized. To this end, a mesocosm study was conducted with a focus on characterizing zooplankton community responses. A single pulse application of thiamethoxam (0, 25, 50, 100, 250, and 500 µg/L; n = 3) was applied to experimental systems and monitored for 8 weeks. The mean half-life of thiamethoxam among the different treatments was 3.7 days in the water column with concentrations of <0.8 µg/L in the majority of mesocosms by 56 days. Principal response curve analysis did not show any significant concentration-dependent differences in the zooplankton community among treatments over the course of the study. The minimum detectable difference (MDD%) values for abundance of potentially sensitive arthropod taxa (nauplius larvae, cyclopoid copepods) allowed the detections from controls as low as 42 and 59% effect, respectively. The MDD% values for total abundance of zooplankton (including the potentially less sensitive taxonomic group of Rotifera) allowed the detection from controls as low as 41% effect. There were no statistically significant differences in zooplankton abundance or diversity between control and treated mesocosms at the end of the study. There were also no statistically significant differences for individual taxa that were sustained between sampling points, or manifested as a concentration-response. We conclude that acute exposure to thiamethoxam at environmentally relevant concentrations (typically ng/L) likely does not represent a significant adverse ecological risk to wetland zooplankton community abundance and structure.

Bioaccumulation of sediment-associated substituted phenylamine antioxidants in Tubifex tubifex and Lampsilis siliquoidea.

Substituted phenylamine antioxidants (SPAs) are additives in a variety of commercial polymers (e.g., lubricants, plastics, etc.). Based on their physicochemical properties, if SPAs were to enter an aquatic system, they would likely partition into sediment and have the capacity to bioaccumulate in biota. This study investigated the potential of four sediment-associated SPAs, diphenylamine (DPA), N-phenyl-1-naphthalene (PNA), N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA), and 4,4'-methylene-bis[N-sec-butylaniline] (MBA) to accumulate in the tissues of freshwater mussels (Lampsilis siliquoidea) and oligochaete worms (Tubifex tubifex). Mussels and worms were exposed to sediment spiked with individual SPAs for 28 d. The concentration of SPAs was measured in the gill, gonad, and remaining viscera of the mussels and entire body of the worms. The majority of biota-sediment accumulation factors (28-d BSAFs) for the different tissues of mussels were < 1. The highest concentrations of SPAs were consistently observed in the gill tissue of mussels relative to the gonad and viscera. The 28-d BSAFs for DPPDA and MBA for worms were < 1, and for DPA and PNA, they ranged from 0.38-2.13 and 1.54-33.24, respectively. The higher 28-d BSAFs observed for worms compared to mussels were likely because worms are endobenthic and feed on sediment-associated organic matter. PNA and DPPDA have similar octanol-water partition coefficients (Kow) but greater 28-d BSAFs were observed for PNA compared to DPPDA for both species. This observation provides evidence that biota may be able to metabolize and/or excrete SPAs with similar physicochemical properties at considerably different rates. The 28-d BSAFs observed for sediment-associated SPAs are lower than those typically required for a chemical to be classified as bioaccumulative.

Evidence of citation bias in the pesticide ecotoxicology literature.

As scientists, we are tasked with letting evidence guide our conclusions. In the world of pesticides this takes on added importance as the data can influence ecological and human health outcomes and regulations, and even the manner in which we grow food. Yet, there seems to be a reticence to engage with the totality of the pesticide ecotoxicology literature, especially papers that report few or no effects or low risk to non-target organisms. We suspected that these studies would have fewer citations than studies that report significant effects or risk for the same compound, and this would be unrelated to the strength of the study, e.g., high quality studies with few or no effects would be cited less frequently than weaker studies that reported effects. To investigate this, we examined a subset of literature around the herbicide atrazine. We found that papers reporting an effect had significantly more citations per year than those that did not (p < 0.05). There was no significant relationship between the strength of the study and number of citations, but a general trend for weaker studies to have greater number of citations. The impact factor of journals was not positively correlated with the strength of the study methods, but studies that reported effects were published in journals with a greater mean impact factor than those that reported no effects (p < 0.05). This analysis reveals evidence of citation bias within the pesticide ecotoxicology literature, as well as bias by journals to publish studies that report effects, regardless of study quality.