Epibenthic Amphipod Research Articles

Development of insecticide resistance in Hyalella azteca

Hyalella azteca are epibenthic amphipods that have developed resistance to pyrethroid and organophosphate insecticides due to single amino acid substitutions in the voltage-gated sodium channel and the acetylcholinesterase-1 gene, respectively. Aquatic systems are often contaminated with several different types of insecticides, therefore there is a possibility that H. azteca have also developed resistance to other classes of insecticides. The aims of the current study were to verify that pyrethroid- and organophosphate-resistant H. azteca have retained their resistance after being cultured in the absence of selective pressure for 5 years (Escondido Creek population) and 9 years (Mosher Slough population), to determine if these populations have cross-resistance to carbaryl (carbamate) and 4,4′-dichlorodiphenyltrichloroethane (DDT; organochlorine), and determine whether previous field exposure to fipronil (phenylpyrazole) and imidacloprid (neonicotinoid) caused resistance in cultured pyrethroid- and organophosphate-resistant H. azteca populations. Escondido Creek and Mosher Slough H. azteca populations both maintained high tolerances for bifenthrin due to L925I and I936F amino acid substitutions. Resistance was also found for chlorpyrifos in the Escondido Creek and Mosher Slough populations with lower genotype frequencies of the G119S substitution, indicating that additional factors may be responsible for organophosphate resistance in this study. Mosher Slough H. azteca were moderately resistant to DDT, and Escondido Creek and Mosher Slough H. azteca were moderately resistant to carbaryl, suggesting cross-resistance. No differences were observed in acute toxicity values across the three populations of H. azteca for fipronil and imidacloprid, and this is possibly due to the lack of exposure to toxic concentrations of these insecticides in the field and lack of similar modes of action to pyrethroids and organophosphates. Resistance is known to be associated with fitness costs that can place insecticide-resistant populations at risk for decline through decreased survival and reduced fecundity.

Influence of water exchange rates on toxicity and bioaccumulation of hydrophobic organic chemicals in sediment toxicity tests.

In standardized sediment toxicity tests, the applied water exchange methods range from static to flow-through conditions and vary between protocols and laboratories even for the same test species. This variation potentially results in variable chemical exposure, hampering the interpretation of toxicity and bioaccumulation. To address these issues, we performed sediment toxicity tests with a mixture of three polycyclic aromatic hydrocarbons (PAHs) and the freshwater epibenthic amphipod Hyalella azteca as model chemicals and organism, respectively. Five standardized water exchange methods were applied: static, semi-static, or flow-through conditions. By measuring total (Cdiss) and freely dissolved concentrations (Cfree) of PAHs with water sampling and direct immersion solid-phase microextraction methods, respectively, we found that Cdiss in overlying water differed by a factor of up to 107 among water exchange conditions, whereas both Cdiss and Cfree in pore water did not differ by more than a factor of 2.6. Similar survival rates, growth rates, and bioaccumulation of PAHs between water exchange methods suggest that H. azteca was predominantly exposed to pore water rather than overlying water. By applying mechanistic kinetic modeling to simulate spatiotemporal concentration profiles in sediment toxicity tests, we discuss the importance of the water exchange rates and resulting temporal and spatial exposure variability for the extrapolation of laboratory sediment toxicity to field conditions, particularly for chemicals with relatively low hydrophobicity and sediments with low organic carbon content.

Genotoxic effects of silver nanoparticles on a tropical marine amphipod via feeding exposure

Silver nanoparticles (AgNP) are widely used in several applications including as antifouling agents; therefore, they can end up in estuarine and marine environments. These nanoparticles tend to aggregate and to deposit in the sediment, where many organisms feed and reproduce. Parhyale hawaiensis is an epibenthic amphipod globally distributed in tropical zones, and has been considered a potential model for ecotoxicology. The aim of this study was to evaluate genotoxic effects of AgNP and Ag salt via feeding, as P. hawaiensis lives in the sediment where nanoparticles tend to accumulate. Organisms were cultivated in the laboratory, and adults were exposed to food containing both AgNP and Ag salt. We collected hemolymph after different times of exposure, and analysed the hemocytes for nuclear abnormalities (including micronuclei) and DNA damage using the standard alkaline comet assay. Conditions of both assays were developed/optimized to allow their successful application in marine invertebrates. Increased frequencies of micronuclei, nuclear buds and total abnormalities were detected in relation to concentration and time in organisms exposed to AgNP and Ag salt in comparison to the controls. No DNA damage was detected when the alkaline comet assay was applied. After 5 days of exposure, we observed higher micronuclei frequencies in Ag salt treatment in comparison with AgNP. After 13 days, micronuclei frequencies were similar for both silver forms. We believe that the Ag, in its ion form, is causing the mutagenic effect; therefore, more time would be needed for the release of the ion from AgNP, explaining the delayed mutagenic effect.

Effects of temperature and salinity on bioconcentration and toxicokinetics of permethrin in pyrethroid-resistant Hyalella azteca

Recent studies demonstrated pyrethroid resistance associated with voltage-gated sodium channel mutations in populations of the epibenthic amphipod, Hyalella azteca. Resistant populations were able to tolerate and bioconcentrate pyrethroids at concentrations significantly higher than toxic levels for non-resistant populations. In conjunction with elevated bioconcentration potential, environmental alteration particularly as a result of global climate change is anticipated to significantly alter abiotic parameters including temperature and salinity. These changes are expected to influence uptake and biotransformation of contaminants. Thus, the aims of the current study were a) to examine the bioconcentration potential of permethrin in two pyrethroid-resistant clades of H. azteca and b) assess the influence of temperature and salinity changes on toxicokinetic parameters. Two pyrethroid-resistant clades of H. azteca were exposed to 14C-permethrin at three salinities (0.2, 1.0 and 6.0 practical salinity units (PSU)) and temperatures (18, 23 and 28 °C). Tests were conducted for up to 36 h and uptake, elimination and biotransformation rates were calculated. Both populations demonstrated bioconcentration factors (BCFs) between five and seven times greater than published data for non-resistant H. azteca, with significant differences between clades. Calculated BCF values were comparable to field populations of resistant H. azteca, emphasizing the potential for elevated pyrethroid bioconcentration in the natural environment and increased exposure for predators consuming pyrethroid-resistant aquatic invertebrates. Alterations to temperature and salinity had no statistically significant effect on uptake or parent compound half-life in either population, though biotransformation was elevated at higher temperatures in both populations. Salinity had a variable effect between the two populations, with lower BCF values at 1.0 PSU in clade D H. azteca and greater BCFs at 6.0 PSU in clade C H. azteca. This is the first study to demonstrate the potential for future climate scenarios to influence toxicokinetics in pyrethroid-resistant aquatic organisms.

SENSIBILIDAD DE Hyalella curvispina SHOEMAKER, 1942 (CRUSTACEA, AMPHIPODA) AL COBRE, FENANTRENO Y SALINIDAD

Bioassays are a useful tool for detecting possible impacts of pollutants on biota. Hyalella curvispina is an epibenthic amphipod native to South America used to evaluate the toxicity of environmental samples. However, information gaps persist regarding their sensitivity to certain pollutants and their tolerance to salinity, so this research provides new data to characterize their response. In this work, the sensitivity to copper and phenanthrene of Hyalella curvispina was analyzed, as well as its tolerance to salinity. The 96h LC50 obtained for copper was 184 µg Cu / l, while the 10-day LC50 was 68 µg Cu / l. Regarding salinity, the results showed an LC50 96h = 13, the LC50 10 days = 11 and a significant inhibition in growth of 18% compared to the control in organisms exposed to 17.2 salinity. For phenanthrene, no significant lethal response was obtained in any of the applied treatments, which suggests that H. curvispina would have a greater tolerance to phenanthrene than other amphipods, however, these preliminary results should be verified covering higher levels of exposure.

Feeding on epibenthic zooplankton by Long‐tailed Ducks: patch structure, profitability, and food web implications

AbstractModels of the trophic needs and impacts of mobile predators in food web networks typically do not consider interactions between prey dispersion and threshold prey densities needed for profitable foraging. As a result, the biomass of predators that can be supported, or direct and indirect impacts of those predators on other food web components, may be poorly estimated. In Simpson Lagoon on the U.S. Arctic coast, epibenthic amphipods and especially mysid shrimp comprised most of the diets of fish, and of Long‐tailed Ducks (Clangula hyemalis) which strongly dominated prey consumption. Based on mean prey densities and gross estimates of prey biomass consumed, earlier studies estimated that continuous immigration of mysids into the lagoon was needed to support the predators. Comparison of variances from net samples vs. different probability density functions (PDFs) around reported means indicated that prey were not normally distributed, and instead occurred in patches with a skewed and wide range of densities. We used the functional response of captive Long‐tailed Ducks eating mysids, and a foraging energetics model, to identify minimum prey densities these diving ducks needed to feed profitably. Based on PDFs that assumed prey densities in all 1‐m2 cells (patches for single feeding dives on potential mysid swarms) were independent, we simulated prey grids with slightly skewed to highly skewed distributions for the reported sample means. Results showed that the greater the range of patch densities, the smaller the fraction of prey biomass that was profitably available to the ducks, thereby increasing the requisite immigration of mysids. Consumption by ectothermic predators further decreased prey available and advanced the date and increased the amount of mysid influx required. The models also showed that the birds had to seek areas with non‐independent clusters of high‐density feeding patches as prey declined in late summer. Thus, spatially continuous measures of patch structure over larger scales are needed to estimate the fraction of prey that is functionally available. Our study emphasizes that when estimating trophic needs and impacts of mobile endothermic predators, simple means of prey density can be misleading, owing to interactions of prey patch structure with profitability thresholds.

Enhanced trophic transfer of chlorpyrifos from resistant Hyalella azteca to inland silversides (Menidia beryllina) and effects on acetylcholinesterase activity and swimming performance at varying temperatures

Chlorpyrifos, an organophosphate (OP) insecticide, is prevalent in aquatic systems globally and is often implicated in aquatic toxicity during storm events. Chlorpyrifos induces toxicity by inhibition of acetylcholinesterase (AChE) activity, which has been related to alterations to fish swimming performance. Resistance to organophosphate insecticides, including chlorpyrifos, is prevalent in populations of the epibenthic amphipod Hyalella azteca in areas with known OP exposure. Previous studies have demonstrated an elevated bioaccumulation potential of insecticide-resistant prey items, however the potential for trophic transfer of chlorpyrifos from OP-resistant prey items and associated neurotoxic effects in fish predators has not been studied. Consequently, the present study aimed to determine the potential for trophic transfer of chlorpyrifos from OP-resistant H. azteca to a known predator, the inland silverside, Menidia beryllina at two temperatures (18 and 23 °C) to simulate temperature changes associated with global climate change (GCC). Fish were fed either 14C-chlorpyrifos-dosed H. azteca or control animals for 7 d, after which total bioaccumulation, percent parent chlorpyrifos, brain AChE activity and swimming performance (ramp-Ucrit) were determined. Fish fed chlorpyrifos-dosed H. azteca bioaccumulated chlorpyrifos ranging from 29.9 to 1250 ng/g lipid, demonstrating the potential for trophic transfer. Lower bioaccumulation and greater biotransformation were observed in M. beryllina at 23 °C as compared to 18 °C, though this was not statistically significant. A significant 36.5% reduction in brain AChE activity was observed in fish fed chlorpyrifos-dosed H. azteca at 23 °C only, which may be attributed to increased biotransformation of parent chlorpyrifos to more potent AChE-inhibiting metabolites. Dietary chlorpyrifos exposure had no significant effect on swimming performance in M. beryllina, though ramp-Ucrit was significantly increased by 25% at 23 as compared to 18 °C. These findings confirm the potential for trophic transfer of chlorpyrifos from OP-resistant prey to fish predators and the potential for elevated temperatures to exacerbate the neurotoxic effects of chlorpyrifos.

Trophic transfer, bioaccumulation and transcriptomic effects of permethrin in inland silversides, Menidia beryllina, under future climate scenarios.

Global climate change (GCC) significantly affects aquatic ecosystems. Continual use of pyrethroid insecticides results in contamination of these ecosystems and concurrent GCC raises the potential for synergistic effects. Resistance to pyrethroids has been documented in Hyalella azteca, a common epibenthic amphipod and model organism. Resistant H.azteca can bioconcentrate elevated amounts of pyrethroids and represent a threat to consumers via trophic transfer. In the present study, a predator of H.azteca, the inland silverside (Menidia beryllina), was used to examine the impacts of GCC on pyrethroid bioaccumulation via trophic transfer from resistant prey organisms. M.beryllina were fed 14C-permethrin dosed pyrethroid-resistant H.azteca for 14 days at three salinities (6, 13 and 20 practical salinity units (PSU)) and two temperatures (18 and 23°C). Fish were analyzed for total body residues, percent parent compound and percent metabolites. Gene expression in liver and brain tissue were evaluated to assess whether dietary bioaccumulation of permethrin would impact detoxification processes, metabolism, and general stress responses. M.beryllina bioaccumulated significant amounts of permethrin across all treatments, ranging from 39 to 557ngg-1 lipid. No statistically significant effect of temperature was found on total bioaccumulation. Salinity had a significant effect on total bioaccumulation, owing to greater bioaccumulation at 6 PSU compared to 13 and 20 PSU, which may be due to alterations to xenobiotic elimination. Permethrin bioaccumulation and the interaction with temperature and salinity elicited significant transcriptional responses in genes relating to detoxification, growth, development, and immune response. Given the increased prevalence of pesticide-resistant aquatic invertebrates, GCC-induced alterations to temperature and salinity, and the predicted increase in pesticide usage, these findings suggest trophic transfer may play an important role in pesticide bioaccumulation and effects in predatory fish.

Contrasting patterns of genetic differentiation for deep-sea amphipod taxa along New Zealand's continental margins

Intraspecific genetic differentiation reflects processes that underpin biodiversity in deep-sea habitats. We examined the spatial genetic structure of epibenthic amphipod taxa from upper bathyal depths (200–1200 m) along New Zealand's continental margins. Genetic variability in amphipods was assessed using cytochrome c oxidase I (COI) to characterise inter-regional diversity and phylogeographic patterns in the Chatham Rise and Challenger Plateau to the east and west of New Zealand, respectively. Automatic barcode gap discovery methods identified 263 MOTUs, of which 14 were detected in both regions. Population genetic analyses found contrasting patterns of diversity among different MOTUs within and between regions. Overall, a pattern of increasing genetic differentiation (K2P distances) with decreasing depth was observed between regions. Genetic differentiation was highest among MOTUs found at shallow depths, while taxa from deeper waters showed little evidence of divergence. Our results suggest that intraspecific genetic diversity is correlated with amphipod family traits, particularly dispersal abilities inferred from habitat preferences (infaunal, epibenthic or pelagic) and may also be linked to regional geological history. Such insights have important implications for regional-scale patterns of diversity associated with local processes in promoting differentiation among populations of bathyal amphipods on New Zealand's continental margins.

The importance of macroalgae and associated amphipods in the selective benthic feeding of sister rockcod species Notothenia rossii and N. coriiceps (Nototheniidae) in West Antarctica

Studies on feeding selectivity in Antarctic fish with comparison between diet organisms and those available in the wild are scarce. We explored this issue in Notothenia rossii (NOR) and N. coriiceps (NOC) at Potter Cove in summer 2016 to test: (1) their preference among different benthic groups, primarily species of algae and amphipods and (2) differences between these nototheniids given their distinct morphology but their known similar general ecology in the fjords. The methodology included a comparative analysis of benthic organisms identified in the stomachs and those collected on macroalgal beds (Ivlev Index). Benthic amphipods, mainly Gondogeneia antarctica and Cheirimedon femoratus, followed by macroalgae, mainly Palmaria decipiens and Desmarestia spp., were the most important and frequent dietary items (IRI%) for both nototheniids. However, NOC was more herbivorous and fed more intensively on a wider diversity of benthic organisms such as certain algal-associated groups like gastropods and bivalves, whereas NOR fed on a greater proportion of epibenthic amphipods and other epibenthic prey. Although in the last three decades the physiognomy of the inner cove has been changed due to the retreat of the Fourcade Glacier, at our sampling site in the outer cove the abundance and vertical distribution of macroalgae did not show substantial changes compared with those reported in the literature in 1994–1996. Temporal differences in the feeding selectivity of NOC on amphipod species, between contemporary and historical samples, may be explained by variations in the assemblage of the algal-associated epifauna. We suggest factors that may have produced these changes.

The Toxicogenome of Hyalella azteca: A Model for Sediment Ecotoxicology and Evolutionary Toxicology.

Hyalella azteca is a cryptic species complex of epibenthic amphipods of interest to ecotoxicology and evolutionary biology. It is the primary crustacean used in North America for sediment toxicity testing and an emerging model for molecular ecotoxicology. To provide molecular resources for sediment quality assessments and evolutionary studies, we sequenced, assembled, and annotated the genome of the H. azteca U.S. Lab Strain. The genome quality and completeness is comparable with other ecotoxicological model species. Through targeted investigation and use of gene expression data sets of H. azteca exposed to pesticides, metals, and other emerging contaminants, we annotated and characterized the major gene families involved in sequestration, detoxification, oxidative stress, and toxicant response. Our results revealed gene loss related to light sensing, but a large expansion in chemoreceptors, likely underlying sensory shifts necessary in their low light habitats. Gene family expansions were also noted for cytochrome P450 genes, cuticle proteins, ion transporters, and include recent gene duplications in the metal sequestration protein, metallothionein. Mapping of differentially expressed transcripts to the genome significantly increased the ability to functionally annotate toxicant responsive genes. The H. azteca genome will greatly facilitate development of genomic tools for environmental assessments and promote an understanding of how evolution shapes toxicological pathways with implications for environmental and human health.

Contaminant exposure effects in a changing climate: how multiple stressors can multiply exposure effects in the amphipod Hyalella azteca.

Global climate change (GCC) is likely to intensify the synergistic effects between altered physicochemical parameters [of changing habitats] and other anthropogenic threats, such as water pollution, posing increased risks to aquatic biodiversity. As such, it is critical to understand how organisms will respond to changes in water temperature and salinity in the presence of contaminants. We exposed the epibenthic amphipod Hyalella azteca to a 3 × 3 factorial treatment design of three temperatures and three salinities ranging from 12 to 18 °C and 0 to 8 parts per thousand (ppt), respectively, in combination with a low-level environmentally relevant concentration of the pyrethroid insecticide bifenthrin (1 ng/L). Effects on survival and swimming behavior were evaluated after 96 h exposure. Transcription of a select suite of genes was monitored at 24, 48, and 96 h using quantitative polymerase chain reaction (qPCR). Our results not only demonstrate that the changes in salinity and temperature result in negative effects to invertebrate survival, behavior, and gene response, but that the effects were significantly more pronounced in the presence of bifenthrin. This is particularly important since greater thermal fluctuations, changes in timing and extent of glacial melt, and changes in precipitation, could result in H. azteca experiencing lower temperatures at times that coincide with increased spraying of pyrethroids. These environmentally relevant exposures using the standard test species H. azteca provide essential information for understanding effects caused by GCC in conjunction with increasing pesticide use, further highlighting the need to incorporate GCC impacts into risk assessments of contaminants of concern.

Variation in the toxicity of sediment-associated substituted phenylamine antioxidants to an epibenthic (Hyalella azteca) and endobenthic (Tubifex tubifex) invertebrate

Substituted phenylamine antioxidants (SPAs) are produced in relatively high volumes and used in a range of applications (e.g., rubber, polyurethane); however, little is known about their toxicity to aquatic biota. Therefore, current study examined the effects of chronic exposure (28 d) to four sediment-associated SPAs on epibenthic (Hyalella azteca) and endobenthic (Tubifex tubifex) organisms. In addition, acute (96-h), water-only exposures were conducted with H. azteca. Mortality, growth and biomass production were assessed in juvenile H. azteca exposed to diphenylamine (DPA), N-phenyl-1-napthylamine (PNA), N-(1,3-dimethylbutyl)-N’-phenyl-1,4-phenylenediamine (DPPDA), or 4,4’-methylene-bis[N-sec-butylaniline] (MBA). Mortality of adult T. tubifex and reproduction were assessed following exposure to the four SPAs. The 96-h LC50s for juvenile H. azteca were 1443, 109, 250, and >22 μg/L and 28-d LC50s were 22, 99, 135, and >403 μg/g dry weight (dw) for DPA, PNA, DPPDA, and MBA, respectively. Reproductive endpoints for T. tubifex (EC50s for production of juveniles > 500 μm: 15, 9, 4, 3.6 μg/g dw, for DPA, PNA, DPPDA, and MBA, respectively) were an order of magnitude more sensitive than endpoints for juvenile H. azteca and mortality of adult worms. The variation in toxicity across the four SPAs was likely related to the bioavailability of the sediment-associated chemicals, which was determined by the chemical properties of the SPAs (e.g., solubility in water, Koc). The variation in the sensitivity between the two species was likely due to differences in the magnitude of exposure, which is a function of the life histories of the epibenthic amphipod and the endobenthic worm. The data generated from this study will support effect characterization for ecological risk assessment.

Copper Sediment Toxicity and Partitioning during Oxidation in a Flow-Through Flume.

The bioavailability of transition metals in sediments often depends on redox conditions in the sediment. We explored how the physicochemistry and toxicity of anoxic Cu-amended sediments changed as they aged (i.e., naturally oxidized) in a flow-through flume. We amended two sediments (Dow and Ocoee) with Cu, incubated the sediments in a flow-through flume, and measured sediment physicochemistry and toxicity over 213 days. As sediments aged, oxygen penetrated sediment to a greater depth, the relative abundance of Fe oxides increased in surface and deep sediments, and the concentration of acid volatile sulfide declined in Ocoee surface sediments. The total pool of Cu in sediments did not change during aging, but porewater Cu, and Cu bound to amorphous Fe oxides decreased while Cu associated with crystalline Fe oxides increased. The dose-response of the epibenthic amphipod Hyalella azteca to sediment total Cu changed over time, with older sediments being less toxic than freshly spiked sediments. We observed a strong dose-response relationship between porewater Cu and H. azteca growth across all sampling periods, and measurable declines in relative growth rates were observed at concentrations below interstitial water criteria established by the U.S. EPA. Further, solid-phase bioavailability models based on AVS and organic carbon were overprotective and poorly predicted toxicity in aged sediments. We suggest that sediment quality criteria for Cu is best established from measurement of Cu in pore water rather than estimating bioavailable Cu from the various solid-phase ligands, which vary temporally and spatially.

Challenges for using quantitative PCR test batteries as a TIE-type approach to identify metal exposure in benthic invertebrates.

The epibenthic amphipod Melita plumulosa shows unique gene expression profiles when exposed to different contaminants. We hypothesized that specific changes in transcript abundance could be used in a battery of quantitative polymerase chain reaction (qPCR) assays as a toxicity identification evaluation (TIE)-like approach to identify the most relevant stressor in field-contaminated sediments. To test this hypothesis, seven candidate transcriptomic markers were selected, and their specificity following metal exposure was confirmed. The performance of these markers across different levels of added metals was verified. The ability of these transcripts to act as markers was tested by exposing amphipods to metal-contaminated field-collected sediments and measuring changes in transcript abundance via qPCR. For two of the three sediments tested, at least some of the transcriptomic patterns matched our predictions, suggesting that they would be effective in helping to identify metal exposure in field sediments. However, following exposure to the third sediment, transcriptomic patterns were unlike our predictions. These results suggest that the seven transcripts may be insufficient to discern individual contaminants from complex mixtures and that microarray or RNA-Seq global gene expression profiles may be more effective for TIE. Changes in transcriptomics based on laboratory exposures to single compounds should be carefully validated before the results are used to analyze mixtures.

Dissolved and Particulate Copper Exposure Induces Differing Gene Expression Profiles and Mechanisms of Toxicity in the Deposit Feeding Amphipod Melita plumulosa

Uptake of metals via ingestion is an important route of exposure for many invertebrates, and it has been suggested that the toxic response to metals accumulated via food differs from that of metals accumulated via the dissolved phase. To test this hypothesis, the deposit-feeding epibenthic amphipod Melita plumulosa was exposed to nontoxic or reproductively toxic concentrations of copper via the overlying water, via ingestion of sediment, or via a combination of the two. Rates of copper uptake from the two exposure routes were predicted using a biokinetic model. Gene expression profiles were measured via microarray analysis and confirmed via quantitative polymerase chain reaction. Differences in expression profiles were related to the exposure route more than to individual or combined rates of copper uptake. Chitinase and digestive protease transcript expression levels correlated to the copper uptake rate from sediment, rather than from the dissolved phase or combined total uptake rate. Overall, this study supports the hypothesis that metals accumulated via ingestion have a different mode of toxic action than metals taken up from water. Consequently, guidelines that only consider dissolved metal exposure, including equilibrium-partitioning-based guidelines, may underestimate the potential effects from deposited or resuspended metal-contaminated sediments.

Assessment of temporal genetic variability of two epibenthic amphipod species in an eastern Australian estuarine environment and their suitability as biological monitors

Population studies often assume temporally stable and consistent patterns of genetic variability. Violations of this assumption can lead to misrepresentation of the amount and patterns of genetic variability in natural populations, which can be problematic in basic research and environmental monitoring studies that are designed to detect environmental perturbation. We collected two endemic species of amphipods, Melita plumulosa and Melita matilda, in a major eastern Australian waterway between November 2009 and October 2011, and assessed genetic variation at the mitochondrial cytochromec oxidase subunitI locus. Overall, M. plumulosa was found to be more genetically variable than M. matilda. No distinct temporal trends in levels and patterns of genetic variation were identified in either species. These findings, combined with the published results demonstrating that M. plumulosa has greater sensitivity to a range of sediment-bound metals and organic contaminants, suggests it to be an informative species for environmental monitoring purposes.

Assessing mechanisms of toxicant response in the amphipod Melita plumulosa through transcriptomic profiling

This study describes the function of transcripts with altered abundance in the epibenthic amphipod, Melita plumulosa, following whole-sediment exposure to a series of common environmental contaminants. M. plumulosa were exposed for 48h to sediments spiked and equilibrated with the following contaminants at concentrations predicted to cause sublethal effects to reproduction: porewater ammonia 30mgL−1; bifenthrin at 100μgkg−1; fipronil at 50μgkg−1; 0.6% diesel; 0.3% crude oil; 250mgCukg−1; 400mgNikg−1; and 400mgZnkg−1. RNA was extracted and hybridized against a custom Agilent microarray developed for this species. Although the microarray represented a partial transcriptome and not all features on the array could be annotated, unique transcriptomic profiles were generated for each of the contaminant exposures. Hierarchical clustering grouped the expression profiles together by contaminant class, with copper and zinc, the petroleum products and nickel, and the pesticides each forming a distinct cluster. Many of the transcriptional changes observed were consistent with patterns previously described in other crustaceans. The changes in the transcriptome demonstrated that contaminant exposure caused changes in digestive function, growth and moulting, and the cytoskeleton following metal exposure, whereas exposure to petroleum products caused changes in carbohydrate metabolism, xenobiotic metabolism and hormone cycling. Functional analysis of these gene expression profiles can provide a better understanding of modes of toxic action and permits the prediction of mixture effects within contaminated ecosystems.

Slow Avoidance Response to Contaminated Sediments Elicits Sublethal Toxicity to Benthic Invertebrates

Advanced analytical techniques have identified the heterogeneity of sediments in aquatic environments which may impact the exposure of benthic organisms to contaminants. Acute and chronic toxicity associated with short, intermittent exposure to four field-collected contaminated sediments were assessed for the epi-benthic amphipod Melita plumulosa and the harpacticoid copepod Nitocra spinipes. Increasing the duration of exposure caused a decrease in survival of M. plumulosa and N. spinipes during 10-d bioassays. Increasing the frequency of exposure to a total exposure time >96-h resulted in a significant toxicity to M. plumulosa. Reproduction decreased for both species from exposure to contaminated sediment. For M. plumulosa, reproductive effects occurred for shorter exposures than the time taken to sense and avoid contaminant exposure. Thus, while avoidance behaviors may prevent acute lethality, slow responses may not prevent sublethal effects. Exposure of M. plumulosa to contaminated sediment appeared to cause a physiological change in females which reduced fecundity. This study indicates that sediment toxicity methods which utilize static continuous exposures may overestimate the toxicity that would occur at a field location. However, by preventing organisms from avoiding unfavorable sediments, these methods provide a precautionary assessment of possible effects, which is usually the aim of most assessments frameworks.

Influence of bioturbation on bioavailability and toxicity of PAHs in sediment from an electronic waste recycling site in South China

The present study examined the effects of bioturbation by the oligochaete Lumbriculus variegatus on bioavailability and toxicity of sediment-associated polycyclic aromatic hydrocarbons (PAHs) to the oligochaete and the epi-benthic amphipod Hyalella azteca. Various densities of L. variegatus in sediment were used to represent different levels of bioturbation. Total sediment concentration declined with increasing worm density, but bioavailability of PAHs estimated using the biomimetic extractions showed no significant difference among treatments with different worm densities. Alternatively, PAH bioaccumulation by L. variegatus decreased and the growth of the worms was reduced at the highest worm density, which was probably due to overcrowding of organisms, food competition and increasing release of PAHs by bioturbation. Additionally, co-exposure with high density of L. variegatus to contaminated sediment significantly increased PAH accumulation and mortality of H. azteca. The increased toxicity was probably because of the transport of sediment-associated contaminants to sediment surface and water column by the bioturbation by L. variegatus. Overall, the present study showed that bioturbation may alter the toxicity of contaminants in sediment to other organisms, thus the presence of benthic invertebrates and their interactions with the sediment should be considered in future sediment risk assessment.