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Miniaturizing MICREDOX A micro-platform for environmental monitoring - applications to BOD and Toxicity

Outline. Microbial sensorsBOD5 - MICREDOX? BODMiniaturizationBODToxicity. Microbial biosensors. use whole-cellsenvironmental applicationsanalyte typedetect target chemical(s)aggregate e.g. BODselective e.g. priority contaminanteffect typereport biological impact e.g. toxicit

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Miniaturizing MICREDOX A micro-platform for environmental monitoring - applications to BOD and Toxicity

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    1. Miniaturizing MICREDOX® A micro-platform for environmental monitoring - applications to BOD and Toxicity

    2. Outline Microbial sensors BOD5 - MICREDOX® BOD Miniaturization BOD Toxicity

    3. Microbial biosensors use whole-cells environmental applications analyte type detect target chemical(s) aggregate e.g. BOD selective e.g. priority contaminant effect type report biological impact e.g. toxicity

    4. Biochemical oxygen demand (BOD5) BOD - international regulatory index for monitoring organic pollutants in water unknown composite substrate - is experimentally inaccessible Dissolved oxygen is the basis of the analytical signal 5-day duration - precludes active intervention GGA standard BOD5 200 mg O2 consumed per litre 60.5% conversion.BOD - international regulatory index for monitoring organic pollutants in water unknown composite substrate - is experimentally inaccessible Dissolved oxygen is the basis of the analytical signal 5-day duration - precludes active intervention GGA standard BOD5 200 mg O2 consumed per litre 60.5% conversion.

    5. MICREDOX® Mediator more soluble than oxygen (1000 fold). Background reduced mediator is zero and its accumulation is a register of the amount of bioconversion. Can increase microbial cell concentration without risk of co-substrate depletion. Initial application BODMediator more soluble than oxygen (1000 fold). Background reduced mediator is zero and its accumulation is a register of the amount of bioconversion. Can increase microbial cell concentration without risk of co-substrate depletion. Initial application BOD

    6. Miniaturization of assay

    7. MICREDOX® miniaturization

    9. MICREDOX® BOD GGA glucose/glutamic acid simple substrate 1º BOD5 standard unsuitable as rapid BOD standard BOD5 100 mg/l OECD Peptone/meat extract complex substrate 2º BOD5 standard recommended as rapid BOD standard BOD5 185 mg/l

    10. Strategy Multivariate analysis Microbial strain (4) Growth media composition (3) Growth media strength (2) Harvest time (2) 48 factor-level combinations (quadruplicate) Measure OECD and GGA standard responses at each level

    11. Main factor effects on MICREDOX® BOD

    12. Main factor trends

    13. MICREDOX® toxicity Reduced mediator is basis of analytical signal Inhibition measured relative to 2 controls without toxin C1 - no added substrate C2 - fixed amount of added substrateReduced mediator is basis of analytical signal Inhibition measured relative to 2 controls without toxin C1 - no added substrate C2 - fixed amount of added substrate

    14. Attenuation of signal by toxin (E.coli / GGA / 3,5-DCP)

    15. 3,5-dichlorophenol

    16. EC50 3,5-dichlorophenol

    17. Conclusions MICREDOX® is amenable to miniaturisation BOD, toxicity Miniaturization significantly boosts throughput of assay BOD quantitative signal (amount of biodegredation) simple substrates can dominate short term signal Toxicity relative signal EC50’s consistent with literature data selection of measurement range and replication allow for estimation of confidence interval

    18. Thanks

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