Grantee Research Project Results
2018 Progress Report: Development of a larval fish neurobehavior adverse outcome pathway to predict effects of contaminants at the ecosystem level and across multiple ecologically relevant taxa
EPA Grant Number: R835798Title: Development of a larval fish neurobehavior adverse outcome pathway to predict effects of contaminants at the ecosystem level and across multiple ecologically relevant taxa
Investigators: Murphy, Cheryl A. , Garcia-Reyero, Natàlia , Carvan, Michael , Jones, Michael
Institution: Michigan State University , University of Wisconsin - Milwaukee , Mississippi State University
EPA Project Officer: Spatz, Kyle
Project Period: June 1, 2015 through May 31, 2018 (Extended to May 31, 2021)
Project Period Covered by this Report: May 1, 2018 through April 30,2019
Project Amount: $800,000
RFA: Systems-Based Research for Evaluating Ecological Impacts of Manufactured Chemicals (2014) RFA Text | Recipients Lists
Research Category: Chemical Safety for Sustainability
Objective:
Objective(s) of the Research Project:
The overall objective of this project is to advance the adverse outcome pathway framework to predict effects of contaminants with different modes of action on the neurobehavior of larval fish from three different species and to determine what Adverse Outcome Pathways (AOPs) are common between species
Objective 1: Identify genes predictive of neurobehavior toxicity in response to exposure to two different chemicals with different molecular initiating events and modes of action in order to identify neurobehavior AOPs using a reverse engineering approach on zebrafish
Objective 2: Determine the effects of PCB126 and MeHg on gene expression and behavior of the larval stage of two species of ecological relevance (fathead minnow and yellow perch).
Objective 3: Incorporate behavioral effects and transcriptomics data from fathead minnow and yellow perch into an individual-based model (IBM) to predict changes in growth and survival to complete the neurobehavior AOP suitable for ecological risk assessment for MeHg and PCB126.
Objective 4: Define and compare neurobehavioral AOPs between species and contaminants to determine their similarities and to elucidate what kind of information is lost or gained by using a typical laboratory model to inform on environmentally relevant species at the population level.
Progress Summary:
Objective 1 and Objective 2.
We have completed exposures for both contaminants and videotaped behavioral assays for all four fish species: yellow perch, zebrafish, killifish and fathead minnows. The behavioral assay data has been collected and analyzed completely for yellow perch and has recently begun for zebrafish and killifish. The brains for all four species were isolated using our newly developed microdissection technique. The brains of fathead minnows were sent off for RNA extraction and the killifish samples are being prepared for RNA extraction. The RNA was extracted from the yellow perch and zebrafish and was in excellent condition, and the data has been received. Currently we are conducting transcriptomic analysis on yellow perch and zebrafish samples. The manuscript describing the yellow perch genome is still in preparation.
The graduate student (Janice Albers) has completed the QA/QC of the yellow perch assay data and has preliminary results of the modeling of six different behavioral endpoints (Figure 1). Additionally, we continue collecting data on the zebrafish locomotion assays, with 230 of the 1800 Zebrafish tracked and finished QA/QC to follow; 170 zebrafish tracked with QA/QC to follow. Finally, we started collecting data from the zebrafish feeding assays with 4 out of 270 finished.
Species | VMR fish count | Locomotion fish count | Feeding fish count |
Zebrafish | 838 | 1800 | 270 |
Yellow Perch | Not possible | 2440 | 351 |
Killifish | 166 | 277 | 206 |
As an example of the type of data collected from the locomotion assays, we have summarized the Yellow Perch behavioral assay results (preliminary since analysis is still in progress) and do see changes in some behaviors in the differing dose levels and chemicals (Figure 1). Currently, are analyzing the same behavioral endpoints for zebrafish larvae using the same analytical procedure.
Figure 1. Mean trends in response variables in yellow perch larvae behavior from locomotion and feeding assays. Y-axis scales are the same for both chemicals, but some y-axis values are back-transformed for those behavior responses that required transformation to meet normality assumptions. * indicates P-values between 0.05-0.1 for tests between treatment and control groups. Error bars represent the 2.5 and 97.5th percentiles of the estimated population posterior distributions. Grey bars represent the 2.5 and 97.5th percentiles of the estimated predicted individual posterior distributions.
Objective 3
The IBM sensitivity analysis has been finalized and the model and a short version of this analysis was presented at SETAC in Nov 2018. This process included much debugging of the code. The IBM was completed to be generalizable so that it can move easily between species using input files. The model presented at SETAC was completed for yellow perch. In order to finalize the calibration of the perch growth, it was necessary to work with Janice Albers, the graduate student who was analyzing the perch locomotion and feeding videos, to ensure compatibility between the data analysis and the model code. Recently, final perch swimming speed, capture success, handling time, reactive distance and total consumption from behavioral analysis was completed. Future work will be needed to finalize perch calibration with these input files, run the sensitivity analysis and compare the effects of contaminants on perch cohort growth rates and abundance. Once similar files are available for fathead minnow and killifish, these runs will also be finalized and completed.
Other Progress
We have a Co-operative agreement with Dr. Diane Nacci's group at the EPA in Narragansett RI. They have completed parallel experiments in killifish. We are going to analyze the behavior and sequence the brains of these killifish. We have been collaborating with Dr. Roger Nisbet and his co-investigators of an EPA STAR grant funded under the same RFP as this grant, through a National Institute of Mathematical and Biological Synthesis (NIMBioS) working group to bridge our adverse outcome pathway work to dynamic energy budgets. This has potential to be quite transformative because it will mean we can screen thousands of chemical to predict impacts on thousands of species.
Future Activities:
We will complete the data collection from the fathead minnow, zebrafish, and killifish behavior assays, analyze the transcriptomic data from all species, and finish analyzing the behavior data. We will complete a neurobehavior AOP.
Journal Articles on this Report : 4 Displayed | Download in RIS Format
Other project views: | All 26 publications | 9 publications in selected types | All 6 journal articles |
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Ivan LN, Schmitt BR, Rose KA, Riley SC, Bose JB, Murphy CA. Evaluation of the thiamine dose-response relationship for lake trout (Salvelinus namaycush) fry using an individual based model. Journal of Great Lakes Research 2018;44:1392-1404. |
R835798 (2018) R835798 (Final) |
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Mueller EB, Lika K, Nisbet RM, Schultz IR, Casas J, Gergs A, Murphy CA, Nacci D, Watanabe KH. Regulation of reproductive processes with dynamic energy budgets. Functional Ecology 2019; 33(5):819-832. |
R835798 (2018) |
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Murphy CA, Nisbet RM, Antczak P, Garcia-Reyero N, Gergs A, Lika K, Mathews T, Muller EB, Nacci D, Peace A, Remien CH, Schultz IR, Stevenson LM, Watanabe KH. Incorporating suborganismal processes into dynamic energy budget models for ecological risk assessment. Integrated Environmental Assessment and Management 2018;14(5):615-624. |
R835798 (2017) R835798 (2018) R835798 (Final) R835797 (2017) |
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Legradi J, DiPaola C, Kraak M, van der Geest H, Schymanski E, Williams A, Dingemans M, Massei R, Brack W, Cousin X, Begout M-L, van der Ost R, Carion A, Suarez Ulloa V, Silvestre F, Escher B, Engwall M, Nilen G, Keiter S, Pollet D, Waldmann P, Kienle C, Werner I, Haigis AC, Knapen D, Vergauwen L, Spher M, Schulz W, Busch W, Leuthold D, Scholz S, vom Berg C, Basu N, Murphy CA, Lampert A, Kuckelkorn J, Grummt T, Hollert H. An ecotoxicological view on neurotoxicity assessment. Environmental Sciences Europe. 2018; 30; 46. |
R835798 (2018) R835798 (Final) |
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Supplemental Keywords:
transcriptomics, larval fish, fathead minnow, yellow perch, zebrafish, neurobehavior, MeHg, PCBs, Adverse outcome pathways, individual-based models, ecological risk assessment, uncertainty, riskProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.
Project Research Results
- Final Report
- 2019 Progress Report
- 2017 Progress Report
- 2016 Progress Report
- 2015 Progress Report
- Original Abstract
6 journal articles for this project