Claudette Kellar Research Summit August 2016 Paratya australiensis: Their Use as a Bioindicator of Impairment in Freshwater Streams Claudette Kellar Research Summit August 2016 One of CAPIMs aims is to develop new tools to use in a weight of evidence approach to help understand biological impairment in various catchments. One of the tools developed is assessing impacts on shrimp in a field setting. Today I am going to give an overview of how we have used Paratya in catchment studies to understand impacts from various stressors.

Paratya australiensis Common shrimp throughout eastern Australia Important food source for native biota Contributes to the majority of macroinvertebrate biomass Important role in ecosystem processes Detrital decomposition Graze extensively on biofilms So Paratya is common and abundant throughout eastern australia and contributes to the majority of macroinvertebrate biomass. Hence it is an important food source for many of our native biota, particularly fish species. They play an important role in ecosystem processes including break down of leaf litter and other detritus and graze heavily of biofilms, directly influencing the structure of algal and benthic invertebrate communities.

Catchment Studies One way to measure biological stress in situ Part of a battery of tests to assess stream condition and factors causing biological impairment Condition of P. australiensis assessed in two ways: Cage studies: lethal and sub lethal effects In situ collections: sub lethal effects The idea of using shrimp in catchment studies is to try and measure stress in situ in the environment. The advantage of this is that we can look at effects of stressors on individuals directly and while potentially reducing costs of continual sampling of the water to measure contaminants in the water. Measuring biological stress in shrimp is part of a number of tests that is used together to assess stream condition and the stressors causing biological impairment. The co

Sub-lethal effects: Biomarkers Identify causal link between chemical exposure and biological effects Provide evidence of ecosystem impairment An early warning indicator that organisms under stress prior to higher level effects

Enzyme Biomarkers Carboxylesterase (CBE) Found in hepatopancreas of shrimp Involved in detoxification/biotransformation of organic chemicals, such as synthetic pyrethroids, in a range of organisms Biomarker of SP (increased activity) Glutathione S Transferase (GST) Involved in detoxification and oxidative stress responses General biomarker

Methods: Caging Experiment Shrimp collected from reference site Maintained in laboratory in site water for seven days 6-8 shrimp put in each cage: 5 cages per site Transported to the field and left at each site for 7-10 days Collected and survival counted and stored on dry ice for biomarker analysis

Methods: In situ Collection Shrimp collected from each site Aim to get 30 individuals from each site Collected and stored on dry ice for biomarker analysis

Methods: Biomarker Preparation Each shrimp weighed and length measured Hepatopancreas removed and weighed for each shrimp Two individuals pooled for GST activity measurements One individual used for CBE activity measurements

Catchment Study: Identifying the Factors Influencing Aquatic Ecosystem Health in the Upper Jacksons Creek Catchment 2013/2014 Aims: Determine and identify the key stressors impacting the health of aquatic fauna in Jacksons Creek Jacksons Creek Urban Area upstream and downstream of the RWP Cattle Deep Creek Reference Site

Chemistry Results - x Nutrients high throughout catchment Control Urban RWP Farming Reference Jacksons JPS Jacksons JURWP JDRWP JSC Chemistry - x Nutrients high throughout catchment Pollution Prevalent: Jacksons Creek at Prince Street JPS high TPH, Pb, Zn, synthetic pyrethroid insecticides (bifenthrin), herbicides (atrazine, simazine, diuron) RWP and downstream herbicides: simazine and diuron, estrogens)

Results: Caged Shrimp

Cage Shrimp Biomarkers AB A B (4) (5) A (5) (4) Site No difference in CBE Activity although highest at JPS and JDRWP No difference in GST Activity between sites

Shrimp In situ Biomarkers (12) (5) (20) (15) (16) (7) (5) (12) (14) (16) Site Higher CBE Activity at site JPS compared to reference site No difference in GST Activity between sites although highest at JPS

Conclusions Increased CBE activity in in situ shrimp at the urban site JPS Increased CBE activity and GST in cage shrimp at site JPS and DRWP although not significant Paratya australiensis is a useful organism to detect biological impairment in rivers. Increase exposure time in cages – 7 days to 10 days

Shrimp utilised in catchment studies Westernport 2015/2016: Pesticide Sourcing and Aquatic Flora and Fauna Assessment No survival at one site in western contour (highest pesticides) Higher GST at WES 1 (elevated pesticides) compared to reference Results indicate pollution is having an affect on P. australiensis (lethal and sub lethal effects).

Shrimp utilised in catchment studies Broken 2015/2016: Lower Broken Creek and Nine Mile Creek macroinvertebrate assessment Lethal effects observed in cage studies at two sites No difference in GST activity Results indicate pollution is having an affect on P. australiensis.

Bioindicator: Paratya australiensis Both in situ and caged Paratya australiensis provide useful information about the condition of stream Caging experiments: Sensitive enough to see lethal effects at sites Biomarkers are able to provide some stress responses In situ collections Information on the condition of shrimp useful (length and weight)

Future Work Lab exposures on different aged shrimp: toxicants, temperature, flow Set up a culture of Paratya australiensis Investigate the ecology of Paratya australiensis

Temperature Exposure Lab Experiment Aim: To measure GST in shrimp exposed to different temperatures: 10⁰C, 16 ⁰C, 21 ⁰C Shrimp collected from Deep Creek Experiment ran for 10 days

Temperature Exposure Lab Experiment 10 days