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New tools to detect ecological effects of contaminants in estuaries
ARC Linkage project
Partners: Melbourne Water, CSIRO and Metabolomics Australia
Sara Long and Allyson O’Brien
Background
• Contaminants (usually in mixtures) can have major impacts on estuaries
• Current assessment methods are: – Do not accurately measure ecological responses – Identify the most severe threat – Expensive
• New molecular techniques may help with this
Aims
• Develop molecular tools (metabolomics and metabarcoding) for monitoring and assessing pollution stress in estuarine ecosystems
• Validation using field experiments
Estuaries • 81% of Australia’s population
live within 50 km of estuaries • Provide valuable ecosystem
services • Impacted by urban
development (metals & pesticides) and agricultural run off (fungicides, insecticides & herbicides)
Environmental Metabolomics • Small molecular weight molecules • Simultaneously measure 50-1000
metabolites • Holistic • A range of methodologies
• Sensitive • Effects observed at lower concentrations
than other endpoints
• Diagnostic for specific chemicals • Cost effective
Collaboration with Dr Dedreia Tull, Dr Con Kouremonos, David De Souza and Prof Malcolm McConville at Bio21 Institute
Research Questions • To what extent might
different taxa respond in the same way to chemicals?
• Are there any chemical-specific metabolites? Little River Estuary. Source: Georgia Sinclair
Approach
• Snail Nassarius burchardi and worm Simplisetia aequisetis
• Common and abundant in estuarine intertidal mudflats
• Easy to collect and maintain in the laboratory • Exposure to copper, zinc and boscalid • Metabolomics analysis
Comparing metabolite responses between species and metals
Trends in metabolite changes - Zinc
Decreased abundance – Tyrosine – Epinephrine – Normetanephrine – Phenylalanine – Tryptophan – Leucine
Photo : Georgia Sinclair
Conclusions • Two species responded
differently to the same chemicals – Worms more sensitive than snails – Family/traits
• Handful of metabolites responded to specifically to zinc
• Metabolomics promising tool to detect effects of metal exposure in estuarine organisms
Metabarcoding
• DNA metabarcoding OR eDNA • Less commonly used in ecological research • Estuarine sediments • Monitoring and diagnostic tool Collaboration with
Dr. Anthony Chariton and Sarah Stephenson
CSIRO Oceans and Atmosphere
Chariton et al. (2010) Frontiers in Ecology and Environment
Field survey
Mesocosms
Field experiments using mesocosms (experimental units)
O’Brien and Keough 2013
Field experiments Individual and community response measured using metabolomics and
metabarcoding 1. Two pesticides
– Diuron (herbicide) – Boscalid (fungicide)
2. Metal and pesticide – Copper in sediment – Pesticide applied by spraying
Outcomes
• Adaptable experimental approach using mesocosms
• Biologically relevant endpoints (individuals & communities)
• Cost-effective compared to traditional counting and identification approach
• Ecological relevant indicators of change
Future directions & opportunities
• Improving details of each approach. • “Community metabolomics &
metabarcoding”. • Test in estuaries along a pollution gradient. • Working with our industry partner, Melbourne
Water, to make these tools useable.
Acknowledgements CAPIM Georgia Sinclair Gigi Woods Rhianna Boyle Kathryn Hassell Ruther Lee Claudette Kellar Liz Morris Katy Jeppe Vin Pettigrove CAPIM team
Funding
• Australian Research Council
ARC LP project LP140100565 • Melbourne Water: Dr
Rhys Coleman & Dr Melita Stevens
• CSIRO
Metabolomics Australia (Bio21) Komal Kanoija Saravanan Dayalan Sean O’Callaghan