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Oxygen effect on Dehalococcoides viability and biomarker quantification.

Benjamin K Amos¹, Kirsti M Ritalahti, Claribel Cruz-Garcia

¹School of Civil and Environmental Engineering and School of Biology, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, Georgia 30332-0512, USA.

Environmental Science & Technology

|August 30, 2008

Summary

Oxygen exposure inhibits Dehalococcoides bacteria crucial for chlorinated contaminant detoxification. Molecular tools struggle to differentiate viable from inhibited cells, highlighting the need for improved bioremediation monitoring methods.

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Area of Science:

Environmental microbiology
Bioremediation science
Molecular biology

Background:

Dehalococcoides bacteria are vital for detoxifying chlorinated contaminants.
Monitoring bioremediation requires quantifying Dehalococcoides DNA and RNA biomarkers.
Oxygen's impact on Dehalococcoides activity and viability is not fully understood.

Purpose of the Study:

To investigate the effects of dissolved oxygen on Dehalococcoides activity, viability, and biomarker quantification.
To assess the reversibility of oxygen inhibition on Dehalococcoides.
To evaluate the efficacy of current molecular tools (qPCR, RT-qPCR) for monitoring oxygen-stressed Dehalococcoides.

Main Methods:

Batch experiments using a Dehalococcoides consortium (BDI) exposed to varying dissolved oxygen levels (< or = 4 mg/L).
Quantification of Dehalococcoides DNA and RNA biomarkers using qPCR and RT-qPCR.
Reversibility experiments involving oxygen exposure followed by oxygen removal.

Main Results:

Oxygen significantly inhibited reductive dechlorination, leading to incomplete degradation to vinyl chloride (VC).
VC dechlorination to ethene was particularly susceptible to oxygen inhibition.
qPCR showed decreased biomarker gene copies, but could not distinguish viable from nonviable cells.
RT-qPCR detected gene transcripts in non-dechlorinating cultures, indicating a disconnect between transcription and activity.

Conclusions:

Dehalococcoides strains exhibit differential sensitivity to oxygen, with VC-to-ethene degraders being more vulnerable.
Current molecular methods (qPCR, RT-qPCR) are insufficient for accurately assessing Dehalococcoides viability and activity post-oxygen exposure.
Development of enhanced molecular tools is necessary for reliable bioremediation monitoring.