Use of pesticides can have substantial nonlethal impacts on nontarget species, including driving evolutionary change, often with unknown consequences for species, ecosystems, and society. Hyalella ...azteca , a species complex of North American freshwater amphipods, is widely used for toxicity testing of water and sediment and has frequently shown toxicity due to pyrethroid pesticides. We demonstrate that 10 populations, 3 from laboratory cultures and 7 from California water bodies, differed by at least 550-fold in sensitivity to pyrethroids. The populations sorted into four phylogenetic groups consistent with species-level divergence. By sequencing the primary pyrethroid target site, the voltage-gated sodium channel, we show that point mutations and their spread in natural populations were responsible for differences in pyrethroid sensitivity. At least one population had both mutant and WT alleles, suggesting ongoing evolution of resistance. Although nonresistant H. azteca were susceptible to the typical neurotoxic effects of pyrethroids, gene expression analysis suggests the mode of action in resistant H. azteca was not neurotoxicity but was oxidative stress sustained only at considerably higher pyrethroid concentrations. The finding that a nontarget aquatic species has acquired resistance to pesticides used only on terrestrial pests is troubling evidence of the impact of chronic pesticide transport from land-based applications into aquatic systems. Our findings have far-reaching implications for continued uncritical use of H. azteca as a principal species for monitoring and environmental policy decisions.
Applications for silver nanomaterials in consumer products are rapidly expanding, creating an urgent need for toxicological examination of the exposure potential and ecological effects of silver ...nanoparticles (AgNPs). The integration of genomic techniques into environmental toxicology has presented new avenues to develop exposure biomarkers and investigate the mode of toxicity of novel chemicals. In the present study we used a 15k oligonucleotide microarray for Daphnia magna, a freshwater crustacean and common indicator species for toxicity, to differentiate between particle specific and ionic silver toxicity and to develop exposure biomarkers for citrate-coated and PVP-coated AgNPs. Gene expression profiles revealed that AgNO3 and AgNPs have distinct expression profiles suggesting different modes of toxicity. Major biological processes disrupted by the AgNPs include protein metabolism and signal transduction. In contrast, AgNO3 caused a downregulation of developmental processes, particularly in sensory development. Metal responsive and DNA damage repair genes were induced by the PVP AgNPs, but not the other treatments. In addition, two specific biomarkers were developed for the environmental detection of PVP AgNPs; although further verification under different environmental conditions is needed.
Transcriptomics, high-throughput assays, and adverse outcome pathways (AOP) are promising approaches applied to toxicity monitoring in the 21st century, but development of these methods is ...challenging for nonmodel organisms and emerging contaminants. For example, Endocrine Disrupting Compounds (EDCs) may cause reproductive impairments and feminization of male bivalves; however, the mechanism linked to this adverse outcome is unknown. To develop mechanism-based biomarkers that may be linked through an AOP, we exposed Mytilus edulis to 17-alpha-ethinylestradiol (5 and 50 ng/L) and 4-nonylphenol (1 and 100 μg/L) for 32 and 39 days. When mussels were exposed to these EDCs, we found elevated female specific transcripts and significant female-skewed sex ratios using a RT-qPCR assay. We performed gene expression analysis on digestive gland tissue using an M. edulis microarray and through network and targeted analyses identified the nongenomic estrogen signaling pathway and steroidogenesis pathway as the likely mechanisms of action for a putative AOP. We also identified several homologues to genes within the vertebrate steroidogenesis pathway including the cholesterol side chain cleavage complex. From this AOP, we designed the Coastal Biosensor for Endocrine Disruption (C-BED) assay which was confirmed in the laboratory and tested in the field.
•Blue mussels Mytilus edulis were deployed at four locations in Boston Harbor.•We identified transcriptomic biomarkers that correlated with PCBs and PAHs.•We targeted endocrine disrupting compounds ...(EDCs) with the C-BED assay.•An Integrated Biomarker Response (IBR) approach identified sites impacted by EDCs.
Within monitoring frameworks, biomarkers provide several benefits because they serve as intermediates between pollutant exposure and effects, and integrate the responses of contaminants that operate through the same mechanism of action. This study was designed to verify the use of transcriptomic biomarkers developed in our prior work (i.e., Coastal Biosensor of Endocrine Disruption; C-BED assay) on Mytilus edulis and identify additional biomarkers for legacy pollutants. M. edulis were collected from a reference site in Pemaquid, ME, USA and deployed by the Massachusetts Water Resources Authority (MWRA) at locations in and outside Boston Harbor, MA, USA: including (1) Boston Inner Harbor (IH), (2) the current outfall (OS), (3) 1 km away from the current outfall (LNB), and (4) Deer Island (DI), the site where untreated wastewater was formerly discharged into the bay. Differential gene expression was quantified with a high density microarray. Seven genes significantly correlated with whole tissue concentration of PAHs, and six genes significantly correlated with whole body concentrations of PCBs, two groups of legacy contaminants that were elevated at stations IH, OS, and DI. Enrichment analysis indicated that IH mussels had the highest induction of stress response genes, which correlated with the higher levels of contaminants measured at this site. Based on the C-BED assay gene analysis, stations IH and OS exhibited signs of endocrine disruption, which were further confirmed by incorporating the results for the C-BED assay within the Integrated Biomarker Response (IBR) approach. This study successfully demonstrated the potential use of transcriptomic biomarkers within a monitoring program to identify the presence and organismal responses to endocrine disrupting and legacy contaminant classes.
Endocrine-disrupting compounds (EDCs), including 17α-ethinyl estradiol (EE2) and 4-nonylphenol (4-NP), enter coastal environments primarily in effluents of wastewater treatment facilities and have ...become ubiquitous in marine surface waters, sediments, and biota. Although EE2 and 4-NP have been detected in marine shellfish, the kinetics of bioconcentration and their tissue distribution have not been thoroughly investigated. The authors performed bioconcentration and depuration experiments in the blue mussel, Mytilus edulis, with 3.37 nM EE2 (0.999 μg/L) and 454 nM 4-NP (100.138 µg/L). Mussels and seawater were sampled throughout a 38-d exposure and a 35-d depuration period, and 6 tissues were individually assayed. Uptake of EE2 and 4-NP was curvilinear throughout exposure and followed a similar uptake pattern: digestive gland > gill ≥ remaining viscera > gonad > adductor > plasma. Depuration varied, however, with half-lives ranging from 2.7 d (plasma) to 92 d (gill) for EE2 and 15 d (plasma) to 57 d (gill) for 4-NP. An innovative modeling approach, with 3 coupled mathematical models, was developed to differentiate the unique roles of the gill and plasma in distributing the EDCs to internal tissues. Plasma appears pivotal in regulating EDC uptake and depuration within the whole mussel.
•Gene expression and metal tissue concentrations were compared in Mytilus edulis.•Expression levels of several transcripts correlated with metal concentrations.•Transcripts involved in the unfolded ...protein response (UPR) were induced.•Integration of transcriptomics and tissue levels provides insight to toxicity.
Marine biomonitoring programs in the U.S. and Europe have historically relied on monitoring tissue concentrations of bivalves to monitor contaminant levels and ecosystem health. By integrating ‘omic methods with these tissue residue approaches we can uncover mechanistic insight to link tissue concentrations to potential toxic effects. In an effort to identify novel biomarkers and better understand the molecular toxicology of metal bioaccumulation in bivalves, we exposed the blue mussel, Mytilus edulis L., to sub-lethal concentrations (0.54μM) of cadmium, lead, and a Cd+Pb mixture. Metal concentrations were measured in gill tissues at 1, 2, and 4 weeks, and increased linearly over the 4 week duration. In addition, there was evidence that Pb interfered with Cd uptake in the mixture treatment. Using a 3025 sequence microarray for M. edulis, we performed transcriptomic analysis, identifying 57 differentially expressed sequences. Hierarchical clustering of these sequences successfully distinguished the different treatment groups demonstrating that the expression profiles were reproducible among the treatments. Enrichment analysis of gene ontology terms identified several biological processes that were perturbed by the treatments, including nucleoside phosphate biosynthetic processes, mRNA metabolic processes, and response to stress. To identify transcripts whose expression level correlated with metal bioaccumulation, we performed Pearson correlation analysis. Several transcripts correlated with gill metal concentrations including mt10, mt20, and contig 48, an unknown transcript containing a wsc domain. In addition, three transcripts directly involved in the unfolded protein response (UPR) were induced in the metal treatments at 2 weeks and were further up-regulated at 4 weeks. Overall, correlation of tissue concentrations and gene expression responses indicates that as mussels accumulate higher concentrations of metals, initial stress responses are mobilized to protect tissues. However, given the role of UPR in apoptosis, it serves as an early indicator of stress, which once overwhelmed will result in adverse physiological effects.
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