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Bioactivity Profiling of Chemical Mixtures for Hazard Characterization

Abstract The assessment and regulation of chemical toxicity to protect human health and the environment are done one chemical at a time and seldom at environmentally relevant concentrations. However, chemicals are found in the environment as mixtures, and their toxicity is largely unknown. Understanding the hazard posed by chemicals within the mixture is critical to enforce protective measures. Here, we demonstrate the application of bioactivity profiling of environmental water samples using the sentinel and ecotoxicology model species Daphnia to reveal the biomolecular response induced by exposure to real-world mixtures. We exposed a Daphnia strain to 30 sampled waters of the Chaobai River and measured the gene expression response profiles. Using a multiblock correlation analysis, we establish correlations between chemical mixtures identified in 30 water samples with gene expression patterns induced by these chemical mixtures. We identified 80 metabolic pathways putatively activated by mixtures of inorganic ions, heavy metals, polycyclic aromatic hydrocarbons, industrial chemicals, and a set of biocides, pesticides, and pharmacologically active substances. Our data-driven approach discovered both known bioactivity signatures with previously described modes of action and new pathways linked to undiscovered potential hazards. This study demonstrates the feasibility of reducing the complexity of real-world mixture toxicity to characterize the biomolecular effects of a defined number of chemical components based on gene expression monitoring of the sentinel species Daphnia.

Engineering↗

Sequim Campus Master Plan

Based in Richland, Washington, Pacific Northwest National Laboratory (PNNL) is one of ten U.S. Department of Energy (DOE) Office of Science (SC) national laboratories. PNNL is DOE’s premier chemistry, earth sciences, and data analytics laboratory, delivering vital mission impacts in energy resiliency and national security. PNNL has 19 core capabilities, each a powerful combination of world-class staff, state-of-the- art equipment, and mission-ready facilities. These capabilities represent a collective set of skills and a body of world-leading scientific and engineering work that provides exceptional value and mission delivery to DOE, the National Nuclear Security Administration (NNSA), the U.S. Department of Homeland Security (DHS), and other federal agencies and industry through strategic partnership projects on a not-to-interfere basis. Currently operated by Battelle Memorial Institute, PNNL occupies more than 2.3 million gross square feet (GSF) of buildings and employs approximately 4,700 staff members. PNNL has implemented a rolling 10-year campus strategy, initiated in fiscal year 2013, designed to deliver the foundation for the multidisciplinary science and engineering expertise, equipment, instrumentation, facilities, and infrastructure required to continue providing exceptional value and mission delivery. This strategy is incorporated into PNNL’s annual Laboratory Plan, which serves as DOE-SC’s equivalent of the Five- Year Site Plan required by DOE Order 430.1C Chg. 1, Real Property Asset Management. The PNNL Sequim Campus is located in Clallam County, just outside of Sequim, Washington. PNNL Sequim builds upon a rich history of research related to marine and coastal resources, environmental chemistry, water resources modeling, ecotoxicology, biotechnology, and national security. Facilities at the PNNL Sequim Campus were designed to take advantage of two unique locational assets: the pristine coastal ocean water in Sequim Bay and the region’s exceptionally clean air. Research facilities on the waterfront are supplied with up to 200 gallons per minute (gpm) of seawater and/or groundwater that enables research on aquatic systems ranging from fully seawater to fresh water, with an onsite water treatment system that removes all biological and hazardous chemical residues prior to discharge. Multiple large seawater tanks outside the waterfront laboratories, along with a boat ramp, pier, and floating dock, enable lab-to-field research. The uplands facility takes advantage of the amazing air quality and is built and operated to enable ultratrace chemical/radiological analytical chemistry. This combination of capabilities is not available at any other marine laboratory in the nation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Stoichiometric Ecotoxicology for a Multisubstance World

Nutritional and contaminant stressors influence organismal physiology, trophic interactions, community structure, and ecosystem-level processes; however, the interactions between toxicity and elemental imbalance in food resources have been examined in only a few ecotoxicity studies. Integrating well-developed ecological theories that cross all levels of biological organization can enhance our understanding of ecotoxicology. In the present article, we underline the opportunity to couple concepts and approaches used in the theory of ecological stoichiometry (ES) to ask ecotoxicological questions and introduce stoichiometric ecotoxicology, a subfield in ecology that examines how contaminant stress, nutrient supply, and elemental constraints interact throughout all levels of biological organization. This conceptual framework unifying ecotoxicology with ES offers potential for both empirical and theoretical studies to deepen our mechanistic understanding of the adverse outcomes of chemicals across ecological scales and improve the predictive powers of ecotoxicology.

60 APPLIED LIFE SCIENCES↗

Extrapolating Contaminant Effects from Individuals to Populations: A Case Study on Nanoparticle Toxicity to Daphnia Fed Environmentally Relevant Food Levels

Ecological risk assessment (ERA) is charged with assessing the likelihood a chemical will have adverse environmental or ecological effects. When assessing the risk of a potential contaminant to biological organisms, ecologists are most concerned with the sustainability of populations of organisms, rather than protecting every individual. However, ERA most commonly relies on data on the effect of a potential contaminant on individuals because these experiments are more feasible than costly population-level exposures. In this work, we address the challenge of extrapolating these individual-level results to predict population-level effects. Previous per-capita population growth rate estimates calculated from individual-level exposures of Daphnia pulicaria to silver nanoparticles (AgNPs) at different food rations predict a critical daily food requirement for daphnid populations exposed to 200 μg/L AgNPs to avoid extinction. To test this, we exposed daphnid populations to the same AgNP concentration at three different food inputs, with the lowest ration close to the extinction threshold predicted from data on individuals. The two populations with the higher food inputs persisted, and the population with the lowest food input went extinct after 50 days but did persist through two generations. Further, we demonstrate that we can extrapolate between these levels of biological organization by parameterizing an individual-level biomass model with data on individuals’ response to AgNPs and using these parameters to predict the outcome for control and AgNP-exposed populations. Key to successful extrapolation is careful modeling of temporal changes in resource density, driven by both the experimental protocols and feedback from the consumer. The implication for ecotoxicology is that estimates of extinction thresholds based on studies of individuals may be reliable predictors of population outcomes, but only with careful treatment of resource dynamics.

54 ENVIRONMENTAL SCIENCES↗

Relationships between maternally-transferred mercury and hatchling development, behavior, and survival in the American alligator ( Alligator mississippiensis )

Mercury is a toxic and pervasive environmental contaminant that can be transferred from mother to offspring during development. Consequences of maternally-transferred mercury have been observed in vertebrate taxa, including reduced clutch viability, reduced offspring size, and behavioral alterations. These sublethal effects have been assumed to decrease survivorship, though this is seldom assessed. Here, we examined how maternally-transferred mercury interacts with incubation temperature to influence reproductive success, offspring behavior, and subsequent survival in the American alligator (Alligator mississippiensis). We collected nine clutches of eggs from a mercury contaminated reservoir on the Savannah River Site, South Carolina, and incubated eggs at either female- or male-promoting temperatures. Clutch-averaged mercury in egg yolk was high relative to other studies in crocodilians and ranged from 0.248 to 0.554 ppm compared to 0.018–0.052 ppm at a site with low levels of mercury contamination; mercury levels in hatchling blood ranged from 0.090 to 0.490 ppm ($\overline{x}$= 0.240 ppm, n = 158). We found few, mostly negligible correlations between life history traits and mercury but noted a positive relationship with egg mass, possibly mediated by correlated maternal effects such as resource provisioning. Incubation temperature exerted strong effects on hatchling phenotypes, with warmer, male-promoting temperatures producing larger and bolder hatchlings. Presumptive females, produced from cooler incubation temperatures, spent more time in warm areas during behavior trials. Hatchlings were released 10–15 days post-hatch and surveyed over eight months to assess survival. Survivorship was positively correlated with hatchling size and negatively correlated with proportional time spent in warm areas. Presumptive females had much lower survival, and overall survivorship for the eight-month period was 0.185–0.208, depending on the modelling approach. Finally, our study suggests that, within the range of concentrations we observed, incubation temperature has a stronger effect on offspring behavior and survival than maternally-transferred mercury pollution in American alligators.

59 BASIC BIOLOGICAL SCIENCES↗