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Understanding the Effects of Environmental Stress from the Cell to the Community Ken M. Jeffries and Richard E. Connon What is environmental stress? • Environmental disturbances caused by one or more stressors. – Stressors cause stress. • Types of stressors: – Natural (e.g., temperature, salinity, predation, disease). – Non-natural (e.g., pollution, dams, fishing). • Disrupts normal processes (i.e., homeostasis). – Organism responds to the stressor(s). • Measurable responses can occur at various levels of biological organism. Different Levels of Biological Organization Ecosystem Effects - Productivity, food webs, ecosystem services Community Effects - Species diversity, predator-prey interactions Population Effects -Abundance, sex-ratios Whole Organism Effects -Mortality, metabolic rates Tissue Effects -Gonad growth, development Cellular and Molecular Effects -Proteins, gene expression, “omics” Modified from: Geist (2011) Ecological Indicators 11: 1507-1516. Ecosystem Effects - Productivity, food webs, ecosystem services Community Effects - Species diversity, predator-prey interactions Population Effects -Abundance, sex-ratios Mesocosms Realistic exposures in “environmental” setting. Long-term community effects (direct/indirect) of multiple species. H. azteca Abundance Determine ecologically-relevant responses over a long exposure period!! p < 0.05 Exposures to a mixture of 3 pesticides (chlorpyrifos, permethrin, lambda cyhalothrin) Applied at concentrations that are equally toxic as determined for hyalella and chironomids. S. Hasenbein, Ph.D. thesis, in progress Ecosystem Effects - Productivity, food webs, ecosystem services Mesocosms Tolerance Population Effects -Abundance, sex-ratios Sensitivity Community Effects - Species diversity, predator-prey interactions S. Hasenbein, Ph.D. thesis, in progress Mortality (e.g., 24 h, 96 h, 14 d). Whole Organism Effects -Mortality, metabolic rates Tissue Effects -Gonad growth, development Control Behavior > (ammonia exposure). Exposed Behavior (ammonia exposure). Control (normal) Exposed (aberrant) Delta Smelt: thermal tolerance 30 (B) A Critical Thermal Maximum (°C) a B a a a 28 26 24 Acute temperature tolerance decreases with age!! B,C C b b b b D 25.4°C Acclimation Temperature (°C) Low Medium High Larval cat.temp high medium low Late-Larval Swanson et al. (2000) Oecologia 123: 384-390 c Juvenile Adult PS-Adult Ontogenetic Stage Age Komoroske et al. (2014) Conservation Physiology 2: cou008 Pesticide exposure (Bifenthrin) Egg protein Log2 Fold-change Reproduction Tissue Effects -Gonad growth, development Cellular and Molecular Effects -Proteins, gene expression, “omics” Brander et al., unpublished data Case Study: Effects of Ibuprofen on Inland Silverside • Often detected in municipal wastewater effluent. • Ibuprofen is one of the most commonly detected pharmaceuticals in the environment. – May have unknown consequences on fish populations. • Emerging contaminant of concern in California. • Ibuprofen is cleared from the body quickly and much of the dose is not used. • Continually reintroduced into aquatic systems. – “Pseudo” persistent in aquatic systems. Inland Silverside • Silverside species are abundant in estuaries throughout North America. – US EPA approved estuarine species for toxicity testing (inland silverside). – Can be raised and spawned in the laboratory. – Useful species for monitoring the effects of contaminants in estuaries. • Attempt to understand how ibuprofen affects inland silversides at multiple levels of biological organization. Reproductive Effects Gonad size Genomics!!!! • Genomics revolution. • Studying biology at genome-wide scales. – Many types of “omics” research. – LOTS of data! • Human genome project. – Map the genome. • 1990 – 2003. • Only take a couple of months now! • Recent abalone die off on CA coast. Genomics football analogy: you heard it here first…. Microarrays!!!! • Microarrays are a tool to screen the expression of thousands of genes at the same time. - “Omics” technology. - Understand biological pathways involved in responses to stressors (football analogy). - Finding unexpected results! • Sequenced molecular information for inland silverside. • Sequences used to develop a species-specific microarray. • 14,393 unique genes (in triplicate) this custom designed microarray. Different Levels of Biological Organization Ecosystem Effects - Productivity, food webs, ecosystem services Community Effects - Species diversity, predator-prey interactions Population Effects -Abundance, sex-ratios Whole Organism Effects -Mortality, metabolic rates Tissue Effects -Gonad growth, development Future Cellular and Molecular Effects Directions -Proteins, gene expression, “omics” Modified from: Geist (2011) Ecological Indicators 11: 1507-1516. Summary • Important to evaluate the impacts of exposure at multiple levels of biological organization. • Mesocosms – monitor responses over long periods of time. – Ecologically-relevant. • Whole organism responses – monitor fitness-level consequences. – Reproductive output – may lead to population declines. • Cellular responses – very sensitive, mechanistic effects. – Understand why responses happen. • Combining these approaches can help us understand complex environmental problems relevant to Northern California. – e.g., ibuprofen case study. Acknowledgements • Connon Lab. – – – – – Bryan Cole Lisa Komoroske Matthias Hasenbein Simone Hasenbein Jen Truong • Nann Fangue Lab. – Dennis Cocherell • Susanne Brander Lab. – Bethany DeCourten • Stephanie Fong, Valerie Connor • UC Davis Aquatic Toxicology Laboratory. – Linda Deanovic, Marie Stillway. • Delta Science Postdoctoral Fellowship. • California Fish & Wildlife. • California Department of Water Resources. • California Department of Pesticide Regulation. Delta Stewardship Council