Toxicokinetics and Particle Number-Based Trophic Transfer of a Metallic Nanoparticle Mixture in a Terrestrial Food Chain.

Herein, we investigated to which extent metallic nanoparticles (MNPs) affect the trophic transfer of other coexisting MNPs from lettuce to terrestrial snails and the associated tissue-specific distribution using toxicokinetic (TK) modeling and single-particle inductively coupled plasma mass spectrometry. During a period of 22 days, snails were fed with lettuce leaves that were root exposed to AgNO3 (0.05 mg/L), AgNPs (0.75 mg/L), TiO2NPs (200 mg/L), and a mixture of AgNPs and TiO2NPs (equivalent doses as for single NPs). The uptake rate constants (ku) were 0.08 and 0.11 kg leaves/kg snail/d for Ag and 1.63 and 1.79 kg leaves/kg snail/d for Ti in snails fed with NPs single- and mixture-exposed lettuce, respectively. The elimination rate constants (ke) of Ag in snails exposed to single AgNPs and mixed AgNPs were comparable to the corresponding ku, while the ke for Ti were lower than the corresponding ku. As a result, single TiO2NP treatments as well as exposure to mixtures containing TiO2NPs induced significant biomagnification from lettuce to snails with kinetic trophic transfer factors (TTFk) of 7.99 and 6.46. The TTFk of Ag in the single AgNPs treatment (1.15 kg leaves/kg snail) was significantly greater than the TTFk in the mixture treatment (0.85 kg leaves/kg snail), while the fraction of Ag remaining in the body of snails after AgNPs exposure (36%) was lower than the Ag fraction remaining after mixture exposure (50%). These results indicated that the presence of TiO2NPs inhibited the trophic transfer of AgNPs from lettuce to snails but enhanced the retention of AgNPs in snails. Biomagnification of AgNPs from lettuce to snails was observed in an AgNPs single treatment using AgNPs number as the dose metric, which was reflected by the particle number-based TTFs of AgNPs in snails (1.67, i.e., higher than 1). The size distribution of AgNPs was shifted across the lettuce-snail food chain. By making use of particle-specific measurements and fitting TK processes, this research provides important implications for potential risks associated with the trophic transfer of MNP mixtures.

Trophic transfer of Cu nanoparticles in a simulated aquatic food chain.

The goal of the current study was to quantify the trophic transfer of copper nanoparticles (CuNPs) in a food chain consisting of the microalga Pseudokirchneriella subcapitata as the representative of primary producer, the grazer Daphnia magna, and the omnivorous mysid Limnomysis benedeni. To quantify the size and number concentration of CuNPs in the biota, tissue extraction with tetramethylammonium hydroxide (TMAH) was performed and quantification was done by single particle inductively coupled plasma mass spectrometry (sp-ICP-MS). The bioconcentration factor (BCF) of the test species for CuNPs varied between 102 - 103 L/kg dry weight when expressing the internal concentration on a mass basis, which was lower than BCF values reported for Cu2+ (103 - 104 L/kg dry weight). The particle size of CuNPs determined by sp-ICP-MS ranged from 22 to 40 nm in the species. No significant changes in the particle size were measured throughout the food chain. Moreover, the measured number of CuNPs in each trophic level was in the order of 1013 particles/kg wet weight. The calculated trophic transfer factor (mass concentration basis) was > 1. This indicates biomagnification of particulate Cu from P. subcapitata to L. benedeni. It was also found that the uptake of particulate Cu (based on the particle number concentration) was mainly from the dietary route rather than from direct aqueous exposure. Furthermore, dietary exposure to CuNPs had a significant effect on the feeding rate of mysid during their transfer from daphnia to mysid and from alga through daphnia to mysid. This work emphasizes the importance of tracing the particulate fraction of metal-based engineered nanoparticles when studying their uptake and trophic transfer.

Trophic Transfer and Toxicity of (Mixtures of) Ag and TiO2 Nanoparticles in the Lettuce-Terrestrial Snail Food Chain.

The increasing application of biosolids and agrochemicals containing silver nanoparticles (AgNPs) and titanium dioxide nanoparticles (TiO2NPs) results in their inevitable accumulation in soil, with unknown implications along terrestrial food chains. Here, the trophic transfer of single NPs and a mixture of AgNPs and TiO2NPs from lettuce to snails and their associated impacts on snails were investigated. Both AgNPs and TiO2NPs were transferred from lettuce to snails with trophic transfer factors (defined as the ratio of the Ag/Ti concentration in snail tissues to the Ag/Ti concentration in lettuce leaves) of 0.2-1.1 for Ag and 3.8-47 for Ti. Moreover, the majority of Ag captured by snails in the AgNP-containing treatments was excreted via feces, whereas more than 70% of Ti was distributed in the digestive gland of snails in the TiO2NP-containing treatments. Additionally, AgNP-containing treatments significantly inhibited the activity of snails, while TiO2NP-containing treatments significantly reduced feces excretion of snails. Furthermore, the concurrent application of AgNPs and TiO2NPs did not affect the biomagnification and distribution patterns of Ag and Ti in snails, whereas their co-existence exhibited more severe inhibition of the growth and activity of snails than in the case of applying AgNPs or TiO2NPs alone. This highlights the possibility of nanoparticle transfer to organisms of higher trophic levels via food chains and the associated risks to ecosystem health.

Mummichog (Fundulus heteroclitus) are less sensitive to 17α-ethinylestradiol (EE2) than other common model teleosts: A comparative review of reproductive effects.

The environmental estrogen 17α-ethinylestradiol (EE2) will depress or completely inhibit egg production in many common model teleosts at low concentrations (≤0.5 ng/L; Runnalls et al., 2015). This inhibition is not seen in the estuarine killifish, or mummichog (Fundulus heteroclitus), even when exposed to 100 ng/L EE2. This relative insensitivity to EE2 exposure indicates species-specific mechanisms for compensating for exogenous estrogenic exposure. This review compares various reproductive responses elicited by EE2 in mummichog to other common model teleosts, such as zebrafish (Danio rerio) and fathead minnow (Pimephales promelas), identifying key endpoints where mummichog differ from other studied fish. For example, EE2 accumulates primarily in the liver/gall bladder of mummichog, which is different than zebrafish and fathead minnow in which accumulation is predominantly in the carcass. Despite causing species-specific differences in fecundity, EE2 has been shown to consistently induce hepatic vitellogenin in males and cause feminization/sex reversal during gonadal differentiation in larval mummichog, similar to other species. In addition, while gonadal steroidogenesis and plasma steroid levels respond to exogenous EE2, it is generally at higher concentrations than observed in other species. In mummichog, production of 17ß-estradiol (E2) by full grown ovarian follicles remains high; unlike other teleost models where E2 synthesis decreases as 17α,20ß-dihydroxy-4-prenen-3-on levels increase to induce oocyte maturation. New evidence in mummichog indicates some dissimilarity in gonadal steroidogenic gene expression responses compared to gene expression responses in zebrafish and fathead minnow exposed to EE2. The role of ovarian physiology continues to warrant investigation regarding the tolerance of mummichog to exogenous EE2 exposure. Here we present a comprehensive review, highlighting key biological differences in response to EE2 exposure between mummichog and other commonly used model teleosts.

Evaluating the environmental impacts of dietary recommendations.

Dietary choices drive both health and environmental outcomes. Information on diets come from many sources, with nationally recommended diets (NRDs) by governmental or similar advisory bodies the most authoritative. Little or no attention is placed on the environmental impacts within NRDs. Here we quantify the impact of nation-specific NRDs, compared with an average diet in 37 nations, representing 64% of global population. We focus on greenhouse gases (GHGs), eutrophication, and land use because these have impacts reaching or exceeding planetary boundaries. We show that compared with average diets, NRDs in high-income nations are associated with reductions in GHG, eutrophication, and land use from 13.0 to 24.8%, 9.8 to 21.3%, and 5.7 to 17.6%, respectively. In upper-middle-income nations, NRDs are associated with slight decrease in impacts of 0.8-12.2%, 7.7-19.4%, and 7.2-18.6%. In poorer middle-income nations, impacts increase by 12.4-17.0%, 24.5-31.9%, and 8.8-14.8%. The reduced environmental impact in high-income countries is driven by reductions in calories (∼54% of effect) and a change in composition (∼46%). The increased environmental impacts of NRDs in low- and middle-income nations are associated with increased intake in animal products. Uniform adoption of NRDs across these nations would result in reductions of 0.19-0.53 Gt CO2 eq⋅a-1, 4.32-10.6 Gt [Formula: see text] eq⋅a-1, and 1.5-2.8 million km2, while providing the health cobenefits of adopting an NRD. As a small number of dietary guidelines are beginning to incorporate more general environmental concerns, we anticipate that this work will provide a standardized baseline for future work to optimize recommended diets further.

Combined effects of salinity, temperature, hypoxia, and Deepwater Horizon oil on Fundulus grandis larvae.

Oil spills have polluted the marine environment for decades and continue to be a major source of polycyclic aromatic hydrocarbons (PAHs) to marine ecosystems around the globe, for example during the 2010 Deepwater Horizon spill. Although the toxicity of PAHs to fish has been well studied, their effects combined with abiotic stressors are poorly understood. The goal of this study was to describe the combined impacts of crude oil and environmental stressors on fish larvae, a sensitive life stage. Gulf killifish (Fundulus grandis) larvae (<24 h post-hatch) were exposed for 48 h to high energy water accommodated fractions (HEWAF; total PAHs 0-125 ppb) of Macondo oil from the Deepwater Horizon spill under different combinations of environmental conditions (dissolved oxygen 2, 6 ppm; temperature 20, 25, 30 °C; salinity 3, 10, 30 ppt). Even under optimal environmental conditions (25 °C, 10 ppt, 6 ppm) larval survival and development were negatively affected by PAHs, starting with the lowest concentration tested (∼15 ppb). Hypoxia and high temperature each increased the adverse effects of HEWAF on development and mortality. In contrast, salinity had little effect on any of the endpoints measured. Importantly, expression of the detoxifying gene cyp1a was highly induced in PAH-exposed larvae under normoxic conditions, but not under hypoxic conditions, potentially explaining the enhanced toxicity observed under hypoxia. This work highlights the importance of considering how suboptimal environmental conditions can exacerbate the effects of pollution on fish early life stages.

The combined effect of Macondo oil and corexit on sheepshead minnow (Cyprinodon variegatus) during early development.

During the 2010 Deepwater Horizon (DWH), oil spill an estimated 800 million L oil and 7.9 million L dispersant entered the environment. The potential adverse effects of the oil-dispersant mixture are poorly understood. The aim of this study was to investigate the impacts of this mixture on early development of sheepshead minnow (Cyprinodon variegatus), a small-bodied estuarine fish commonly found in the area affected by the DWH spill. Embryos were exposed to a chemically enhanced water accommodated fraction (CEWAF; 10:1 mixture of Macondo oil; Corexit 9500) for 48 hr, after which organisms were maintained in uncontaminated water for an additional 8 days. Impacts were assessed on embryonic (heart rate, development of eye pigmentation, embryonic movement measured) and larval (time to hatch, larval survival, standard length, and cyp1a gene expression) development. No significant alterations were found in survival, time to hatch or cyp1a at the end of the experiment. However, CEWAF induced significant decreases in heart rate of embryos, delayed development of eye pigmentation, reduced embryonic movement, and diminished standard length. These results indicate potential sublethal adverse consequences attributed to CEWAF exposure during early development, even in the absence of maintained cyp1a induction or survival rate, potentially affecting the fitness of organisms later in life.

Harp Seals Do Not Increase Their Call Frequencies When It Gets Noisier.

Some species avoid low-frequency masking by shifting their calls to higher frequencies. We addressed the hypothesis that Pagophilus groenlandicus (harp seals) will make more high-frequency underwater calls to avoid low-frequency conspecific masking as calling rates increase. The spectral shapes at high and low calling rates were compared (after equalizing the broadband amplitudes). There were no significant differences between the spectral shapes. Pagophilus groenlandicus do not alter the proportions of low- and high-frequency calls as it gets noisier. This suggests that they may not shift their calling frequencies when encountering low-frequency, broadband anthropogenic noise.

Participatory hackathon to determine ecological relevant endpoints for a neurotoxin to aquatic and benthic invertebrates.

The aim of this study is twofold: i) to determine innovative yet sensitive endpoints for sulfoxaflor and ii) to develop best practices for innovative teaching in ecotoxicology. To this end, a group of 52 MSc students participated in an environmental hackathon, during which they did creative toxicity testing on 5 freshwater invertebrate species: Daphnia magna, Chironomus riparius, Asellus aquaticus, Lymnaea stagnalis, and Anisus vortex. Involving the students in an active learning environment stimulated increased creativity and productivity. In total, 28 endpoints were investigated, including standard endpoints (e.g., mortality) as well as biomechanistic and energy-related endpoints. Despite high variances in the results, likely linked to the limited lab experience of the students and interpersonal differences, a promising set of endpoints was selected for further investigation. A more targeted follow-up experiment focused on the most promising organism and set of endpoints: biomechanistic endpoints of C. riparius larvae. Larvae were exposed to a range of sulfoxaflor concentrations (0.90-67.2 µg/L) for 21 days. Video tracking showed that undulation and swimming were significantly reduced at 11.1 µg sulfoxaflor/L after 9 days of exposure, and an EC50 = 10.6 µg/L for mean velocities of the larvae in the water phase was found. Biomechanistic endpoints proved much more sensitive than mortality, for which an LC50 value of 116 µg/L was found on Day 9. Our results show that performing a hackathon with students has excellent potential to find sensitive endpoints that can subsequently be verified using more targeted and professional follow-up experiments. Furthermore, utilising hackathon events in teaching can increase students' enthusiasm about ecotoxicology, driving better learning experiences.

Comparing the impact of microplastics derived from a biodegradable and a conventional plastic mulch on plant performance.

Agricultural lands have been identified as plastic sinks. One source is plastic mulches, which are a source of micro- and nano-sized plastics in agricultural soils. Because of their persistence, there is now a push towards developing biodegradable plastics, which are designed to undergo (partial) breakdown after entering the environment. Yet, limited research has investigated the impacts of both conventional and biodegradable plastics on distinct plants. Moreover, comparisons among studies are difficult due to differences in experimental design. This study directly compares the effects of artificially weathered conventional polyethylene (PE) and starch-based biodegradable polybutylene adipate terephthalate (PBAT) on four food crops, including two monocots (barley, Hordeum vulgare, and wheat, Triticum aestivum L.) and two dicots (carrot, Daucus carota, and lettuce, Lactuca sativa L.). We investigated the effects of environmentally relevant low, medium, and high (0.01 %, 0.1 %, 1 % w/w) concentrations of PE and starch-PBAT blend on seed germination (acute toxicity), and subsequently on plant growth and chlorophyll through a pot-plant experiment (chronic toxicity). Germination of all species was not affected by both plastics. However, root length was reduced for lettuce and wheat seedlings. No other effects were recorded on monocots. We observed a reduction in shoot length and bud wet weight of carrot seedlings for the highest concentration of PE and starch-PBAT blend. Chronic exposure resulted in a significant decrease in shoot biomass of barley and lettuce. Additionally, a positive increase in the number of leaves of lettuce was observed for both plastics. Chlorophyll content was increased in lettuce when exposed to PE and starch-PBAT blend. Overall, adverse effects in dicots were more abundant than in monocots. Importantly, we found that the biodegradable plastic caused more commonly adverse effects on plants compared to conventional plastic, which was confirmed by a mini-review of studies directly comparing the impact of conventional and biodegradable microplastics.

Nano- and microplastics commonly cause adverse impacts on plants at environmentally relevant levels: A systematic review.

Over the last years there has been significant research on the presence and effects of plastics in terrestrial systems. Here we summarize current research findings on the effects of nano- and microplastics (NMPs) on terrestrial plants, with the aim to determine patterns of response and sensitive endpoints. We conducted a systematic review (based on 78 studies) on the effects of NMPs on germination, plant growth and biochemical biomarkers. This review highlights that the majority of studies to date have used pristine polystyrene or polyethylene particles, either in a hydroponic or pot-plant setup. Based on these studies we found that effects on plants are widespread. We noted similar responses between and within monocots and dicots to NMPs, except for consistent lower germination seen in dicots exposed to NMPs. During early development, germination and root growth are more strongly affected compared to shoot growth. NMPs induced similar adverse growth effects on plant biomass and length in the most tested plant species (lettuce, wheat, corn, and rice) irrespective of the polymer type and size used. Moreover, biomarker responses were consistent across species; chlorophyll levels were commonly negatively affected, while stress indicators (e.g., ROS or free radicals) and stress respondents (e.g., antioxidant enzymes) were consistently upregulated. In addition, effects were commonly observed at environmentally relevant levels. These findings provide clear evidence that NMPs have wide-ranging impacts on plant performance. However, as most studies have been conducted under highly controlled conditions and with pristine plastics, there is an urgent need to test under more environmentally realistic conditions to ensure the lab-based studies can be extrapolated to the field.

Species-dependent responses of crop plants to polystyrene microplastics.

Only recently there has been a strong focus on the impacts of microplastics on terrestrial crop plants. This study aims to examine and compare the effects of microplastics on two monocotyledonous (barley, Hordeum vulgare and wheat, Triticum aestivum), and two dicotyledonous (carrot, Daucus carota and lettuce, Lactuca sativa) plant species through two complimentary experiments. First, we investigated the effects of low, medium, and high (103, 105, 107 particles per mL) concentrations of 500 nm polystyrene microplastics (PS-MPs) on seed germination and early development. We found species-dependent effects on the early development, with microplastics only significantly affecting lettuce and carrot. When acutely exposed during germination, PS-MPs significantly delayed the germination of lettuce by 24%, as well as promoted the shoot growth of carrot by 71% and decreased its biomass by 26%. No effect was recorded on monocot species. Secondly, we performed a chronic (21 d) hydroponic experiment on lettuce and wheat. We observed that PS-MPs significantly reduced the shoot growth of lettuce by up to 35% and increased its biomass by up to 64%, while no record was reported on wheat. In addition, stress level indicators and defence mechanisms were significantly up-regulated in both lettuce and wheat seedlings. Overall, this study shows that PS-MPs affect plant development: impacts were recorded on both germination and growth for dicots, and responses identified by biochemical markers of stress were increased in both lettuce and wheat. This highlights species-dependent effects as the four crops were grown under identical conditions to allow direct comparison. For future research, our study emphasizes the need to focus on crop specific effects, while also working towards knowledge of plastic-induced impacts at environmentally relevant conditions.

Polystyrene nanoparticles induce concerted response of plant defense mechanisms in plant cells.

Recent advances in knowledge suggest that micro- and nanoplastics pose a threat to plant health, however, the responses of plants to this stressor are not well-known. Here we examined the response of plant cell defence mechanisms to nanoparticles of commonly used plastic, polystyrene. We used plant cell cultures of widely cultivated plants, the monocots wheat and barley (Triticum aestivum L., Hordeum vulgare L.) and the dicots carrot and tomato (Daucus carota L., Solanum lycopersicum L.). We measured the activities of enzymes involved in the scavenging of reactive oxygen species and nonenzymatic antioxidants and we estimated potential damages in plant cell structures and functioning via lipid peroxidation and DNA methylation levels. Our results demonstrate that the mode of action of polystyrene nanoparticles on plant cells involves oxidative stress. However, the changes in plant defence mechanisms are dependent on plant species, exposure time and nanoplastic concentrations. In general, both monocots showed similar responses to nanoplastics, but the carrot followed more the response of monocots than a second dicot, a tomato. Higher H2O2, lipid peroxidation and lower enzyme activities scavenging H2O2 suggest that tomato cells may be more susceptible to polystyrene-induced stress. In conclusion, polystyrene nanoplastics induce oxidative stress and the response of the plant defense mechanisms involving several chain reactions leading to oxidoreductive homeostasis.

Predicted microplastic uptake through trophic transfer by the short-beaked common dolphin (Delphinus delphis) and common bottlenose dolphin (Tursiops truncatus) in the Northeast Atlantic Ocean and Mediterranean Sea.

Marine mammals can serve as an indicator of ecosystem health, and are likely exposed to significant amounts of microplastics (MPs). In this study we estimated the MP uptake of two odontocetes, the short-beaked common dolphin (Delphinus delphis) and the common bottlenose dolphin (Tursiops truncatus), in the Mediterranean Sea and the Northeast Atlantic. These two species are expected to primarily ingest MPs through trophic transfer. To this end, data was collected on their diet, which was subsequently linked to MP occurrence and abundance in prey families. We estimated that D. delphis ingests 76 MPs/day in the Northeast Atlantic and 164 MPs/day in the Mediterranean, and T. truncatus ingests 36 MPs/day in the Northeast Atlantic and 179 MPs/day in the Mediterranean. This study provides important new predictions on MP exposure in two odontocetes, and opens up new research opportunities on the effect of this exposure on the health of organisms.

The potential use of Atlantic silverside (Menidia menidia) for monitoring estuarine pollution.

Monitoring chronic impacts of nearshore pollution on fish in Atlantic marine systems is challenging due to the mobility and seasonal nature of most habitat use. We evaluated Atlantic silverside (Menidia menidia) as a potential indicator of pollution across a gradient of contaminated sites in the Saint John River Estuary, New Brunswick, Canada. Fish returning from offshore in the spring showed no differences in length, weight, or condition factor between sites, but a graded response in condition factor appeared gradually, consistent with levels of contamination. Gonad size peaked in early June, however no differences in GSI were found among sites. By September, most fish collected were young of the year, with a significant bias towards female juveniles at the exposed site. These results indicate that Atlantic silverside are locally resident during their onshore period, and demonstrates their potential as a sentinel species. Sampling should focus on early June prior to the full moon to ensure maximal exposure periods before spawning, or in late October to evaluate growth as fish grow rapidly prior to offshore movement.

Marine mammals and microplastics: A systematic review and call for standardisation.

Microplastics receive significant societal and scientific attention due to increasing concerns about their impact on the environment and human health. Marine mammals are considered indicators for marine ecosystem health and many species are of conservation concern due to a multitude of anthropogenic stressors. Marine mammals may be vulnerable to microplastic exposure from the environment, via direct ingestion from sea water, and indirect uptake from their prey. Here we present the first systematic review of literature on microplastics and marine mammals, composing of 30 studies in total. The majority of studies examined the gastrointestinal tracts of beached, bycaught or hunted cetaceans and pinnipeds, and found that microplastics were present in all but one study, and the abundance varied between 0 and 88 particles per animal. Additionally, microplastics in pinniped scats (faeces) were detected in eight out of ten studies, with incidences ranging from 0% of animals to 100%. Our review highlights considerable methodological and reporting deficiencies and differences among papers, making comparisons and extrapolation across studies difficult. We suggest best practices to avoid these issues in future studies. In addition to empirical studies that quantified microplastics in animals and scat, ten studies out of 30 (all focussing on cetaceans) tried to estimate the risk of exposure using two main approaches; i) overlaying microplastic in the environment (water or prey) with cetacean habitat or ii) proposing biological or chemical biomarkers of exposure. We discuss advice and best practices on research into the exposure and impact of microplastics in marine mammals. This work on marine ecosystem health indicator species will provide valuable and comparable information in the future.

The influence of hypoxia on the cardiac transcriptomes of two estuarine species - C. variegatus and F. grandis.

Increased nutrient loading has led to eutrophication of coastal shelf waters which has resulted in increased prevalence of persistent hypoxic zones - areas in which the dissolved oxygen content of the water drops below 2 mg/L. The northern Gulf of Mexico, fed primarily by the Mississippi River watershed, undergoes annual establishment of one of the largest hypoxic zones in the world. Exposure to hypoxia can induce physiological impacts in fish cardiac systems that include bradycardia, changes in stroke volume, and altered cardiovascular vessel development. While these impacts have been addressed at the functional level, there is little information regarding the molecular basis for these changes. This study used transcriptomic analysis techniques to interrogate the effects of hypoxia exposure on the developing cardiovascular system in newly hatched larvae of two estuarine species that occupy the same ecological niche - the sheepshead minnow (Cyprinodon variegatus) and the Gulf killifish (Fundulus grandis). Results suggest that while differential gene expression is largely distinct between the two species, downstream impacts on pathways and functional responses such as reduced cardiac hypertrophy, modulation of blood pressure, and increased incidence of apoptosis appear to be conserved. Further, differences in the magnitude of these conserved responses may suggest that the length of embryonic development could impart a level of resiliency to hypoxic perturbation in early life stage fish.

Effects of polycyclic aromatic hydrocarbons and abiotic stressors on Fundulus grandis cardiac transcriptomics.

Following the 2010 Deepwater Horizon oil spill, extensive research has been conducted on the toxicity of oil and polycyclic aromatic hydrocarbons (PAHs) in the aquatic environment. Many studies have identified the toxicological effects of PAHs in estuarine and marine fishes, however, only recently has work begun to identify the combinatorial effect of PAHs and abiotic environmental factors such as hypoxia, salinity, and temperature. This study aims to characterize the combined effects of abiotic stressors and PAH exposure on the cardiac transcriptomes of developing Fundulus grandis larvae. In this study, F. grandis larvae were exposed to varying environmental conditions (dissolved oxygen (DO) 2, 6 ppm; temperature 20, 30 °C; and salinity 3, 30 ppt) as well as to a single concentration of high energy water accommodated fraction (HEWAF) (∑PAHs 15 ppb). Whole larvae were sampled for RNA and transcriptional changes were quantified using RNA-Seq followed by qPCR for a set of target genes. Analysis revealed that exposure to oil and abiotic stressors impacts signaling pathways associated with cardiovascular function. Specifically, combined exposures appear to reduce development of the systemic vasculature as well as strongly impact the cardiac musculature through cardiomyocyte proliferation resulting in inhibited cardiac function and modulated blood pressure maintenance. Results of this study provide a holistic view of impacts of PAHs and common environmental stressors on the cardiac system in early life stage estuarine species. To our knowledge, this study is one of the first to simultaneously manipulate oil exposure with abiotic factors (DO, salinity, temperature) and the first to analyze cardiac transcriptional responses under these co-exposures.

Validation of a refined short-term adult fish reproductive test with improved power for mummichog (Fundulus heteroclitus) to test complex effluents.

Short-term adult fish reproductive tests are widely used to assess the toxicity of chemicals and waste streams. However, these tests often have low power to detect differences in egg production among treatments, due to high variance and small sample sizes, limiting their effectiveness for informing regulatory decisions. A protocol for a fish reproductive test using mummichog (Fundulus heteroclitus) was refined to increase statistical power. Three studies using the original protocol were compared with three studies using the refined protocol. Tank pre-selection and sample size increased the a priori power from 11.2% to 85.7%. After exposure, average power levels were 62.0%, a more than five-fold increase compared to studies that used the original protocol (power of 15.0%). There was a high level of consistency compared to the original protocol; differences >33% in female and male gonad size and egg production could be detected among treatments. This study demonstrates that a refinement process can address shortcomings in short-term adult fish reproductive protocols, creating a solid foundation for further standardization and possible regulatory use.

Quantifying the relative contribution of particulate versus dissolved silver to toxicity and uptake kinetics of silver nanowires in lettuce: impact of size and coating.

Functionalized high-aspect-ratio silver nanowires (AgNWs) have been recognized as one of the most promising alternatives for fabricating products, with their use ranging from electronic devices to biomedical fields. Given concerns on the safety of AgNWs, there is an urgent need to investigate the relation between intrinsic properties of AgNWs and their toxicity. In this study, lettuce was exposed for either 6 or 18 d to different AgNWs to determine how the size/aspect ratio and coating of AgNWs affect the contributions of the dissolved and particulate Ag to the overall phytotoxicity and uptake kinetics. We found that the uncoated AgNW (39 nm diameter × 8.4 µm length) dissolved fastest of all AgNWs investigated. The phytotoxicity, uptake rate constants, and bioaccumulation factors of the PVP-coated AgNW (43 nm diameter × 1.8 µm length) and the uncoated AgNW (39 nm diameter × 8.4 µm length) were similar, and both were higher than that of the PVP-coated AgNW with the larger diameter(65 nm diameter × 4.4 µm length). These results showed that the diameter of the AgNWs predominantly affected toxicity and Ag accumulation in plants. Particulate Ag was found to be the predominant driver/descriptor of overall toxicity and Ag accumulation in the plants rather than dissolved Ag for all AgNWs tested. The relative contribution of dissolved versus particulate Ag to the overall effects was influenced by the exposure concentration and the extent of dissolution of AgNWs. This work highlights inherent particulate-dependent effects of AgNWs in plants and suggests that toxicokinetics should explicitly be considered for more nanomaterials and organisms, consequently providing more realistic input information for their environmental risk assessment.