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InthespiritofGENERA
SharonHook|SeniorEcotoxicologist 7May2012
LANDANDWATER

Rapidsublethalsedimentbioassays

250 m

120 Reproduction, % control l 100 80 60 40 20 0 0 01 0.01 Melita plumulosa Nitocra spinipes 01 0.1 1 Mean SQG-(TV) Quotient (Total metals) 10

4 DAYS DURATION Day 0 Day 4 Day 4 Survival Gravids 5 ml

7 DAYS DURATION Day 11

5 Gravids

Nauplii Copepodytes

Reference: Perez-Landa and Simpson. (2011)

Significantly more sensitive, and twice as rapid, than all previous methods while remaining robust
Simpson,S.L.,Spadaro,D.A.(2011)Performanceandsensitivityofrapidsublethal sediment toxicitytestswiththeamphipodMelita plumulosa andcopepodNitocra spinipes Presentationtitle|Presentername|Page2 EnvironmentalToxicologyandChemistry30,23262334.
5 mm

Algal Growth Inhibition Bioassay

Count cells I Incubation b ti

Culture algae

Add to assay flasks

72 hour later

Flow cytometer

Data p processing g % growth inhibition

Unicellular freshwater alga Chlamydomonas reinhardtii initial cell density 104 cells/ml Test medium: US EPA medium (2002) without EDTA, modified to contain low chloride concentrations concentrations, buffered to pH 7 7.5 5 with 2 mM PIPES, hardness 15 mg/L 72-hour 72 h growth th i inhibition hibiti t test t on nanoparticulate ti l t Ag, A bulk b lk A Ag and dissolved Ag (as AgNO3) Cells were counted daily using flow cytometry
(Franklin et al., 2004)

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Howcanweintegratemodern moleculartechniques?

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Wevedonesomeproofofconceptwork with ithmicroarrays i

In both diatoms and amphipods, see toxicant specific gene expression profiles In amphipods, gene expression profiles change with exposure route

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Wantedtobeabletoapplyadvancesinomictechnologyto localtestspecies But*no* * *apriori sequenceinformationwas available Used U d454pyrosequencing i to t capture t the th t transcriptome i t f forMelita M lit plumulosa,Saccostreaglomerata,Nitschiaclosterium,Chlorella p 12 sp.

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Thankyou y
SharonHook SeniorEcotoxicologist t +61 61297106839 e Sharon.Hook@csiro.au

LANDANDWATER