Skip to main content
SpringerLink
Log in

Assessing the correlation between anaerobic toluene degradation activity and bssA concentrations in hydrocarbon-contaminated aquifer material

  • Original Paper
  • Published:
Biodegradation Aims and scope Submit manuscript

Abstract

The assessment of biodegradation activity in contaminated aquifers is critical to demonstrate the performance of bioremediation and natural attenuation and to parameterize models of contaminant plume dynamics. Real time quantitative PCR (qPCR) was used to target the catabolic bssA gene (coding for benzylsuccinate synthase) and a 16S rDNA phylogenetic gene (for total Bacteria) as potential biomarkers to infer on anaerobic toluene degradation rates. A significant correlation (P = 0.0003) was found over a wide range of initial toluene concentrations (1–100 mg/l) between toluene degradation rates and bssA concentrations in anaerobic microcosms prepared with aquifer material from a hydrocarbon contaminated site. In contrast, the correlation between toluene degradation activity and total Bacteria concentrations was not significant (P = 0.1125). This suggests that qPCR targeting of functional genes might offer a simple approach to estimate in situ biodegradation activity, which would enhance site investigation and modeling of natural attenuation at hydrocarbon-contaminated sites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Allen JP, Atekwana EA, Duris JW, Werkema DD, Rossbach S (2007) The microbial community structure in petroleum contaminated sediments corresponds to geophysical signatures. Appl Environ Microbiol 73:2860–2870

    Article  CAS  PubMed  Google Scholar 

  • Alvarez PJJ, Illman W (2006) Bioremediation and natural attenuation: process fundamentals and mathematical models. In: Schnoor JL, Zehnder A (eds) Environmental science and technology. Wiley, Hoboken, pp 1–608

    Google Scholar 

  • Baelum J, Henriksen T, Hansen HC, Jacobsen CS (2006) Degradation of 4-chloro-2-methylphenoxyacetic acid in top- and subsoil is quantitatively linked to the class III tfdA gene. Appl Environ Microbiol 72:1476–1486

    Article  CAS  PubMed  Google Scholar 

  • Beller HR, Edwards EA (2000) Anaerobic toluene activation by benzylsuccinate synthase in a highly enriched methanogenic culture. Appl Environ Microbiol 66:5503–5505

    Article  CAS  PubMed  Google Scholar 

  • Beller HR, Spormann AM (1997) Anaerobic activation of toluene and o-xylene by addition to fumarate in denitrifying strain T. J Bacteriol 179:670–676

    CAS  PubMed  Google Scholar 

  • Beller HR, Kane SR, Legler TC, Alvarez PJJ (2002) A real-time polymerase chain reaction method for monitoring anaerobic, hydrocarbon-degrading bacteria based on a catabolic gene. Environ Sci Technol 36:3977–3984

    Article  CAS  PubMed  Google Scholar 

  • Beller HR, Kane SR, Legler TC, McKelvie JR, Lollar BS, Pearson F, Balser L, Mackay DM (2008) Comparative assessment of benzene, toluene, and xylene natural attenuation by quantitative polymerase chain reaction analysis of a catabolic gene, signature metabolites, and compound-specific isotope analysis. Environ Sci Technol 42:6065–6072

    Article  CAS  PubMed  Google Scholar 

  • Biegert T, Fuchs G, Heider J (1996) Evidence that anaerobic oxidation of toluene in the denitrifying bacterium Thauera aromatica is initiated by formation of benzylsuccinate from toluene and fumarate. Eur J Biochem 238:661–668

    Article  CAS  PubMed  Google Scholar 

  • Chin K-J, Esteve-Nunez A, Leang C, Lovley DR (2004) Direct correlation between rates of anaerobic respiration and levels of mRNA for key respiratory genes in Geobacter sulfurreducens. Appl Environ Microbiol 70:5183–5189

    Article  CAS  PubMed  Google Scholar 

  • Da Silva MLB, Alvarez PJJ (2004) Enhanced anaerobic biodegradation of benzene–toluene–ethylbenzene–xylene–ethanol mixtures in bioaugmented aquifer columns. Appl Environ Microbiol 70:4720–4726

    Article  CAS  PubMed  Google Scholar 

  • Da Silva MLB, Alvarez PJJ (2008) Exploring the correlation between halorespirer biomarker concentrations and TCE dechlorination rates. J Environ Eng 134:895–901

    Article  CAS  Google Scholar 

  • Fleming JT, Sanseverino J, Sayler GS (1993) Quantitative relationship between naphthalene catabolic gene frequency and expression in predicting PAH degradation in soils at town gas manufacturing sites. Environ Sci Technol 27:1068–1074

    Article  CAS  Google Scholar 

  • Hutchins SR (1997) Effects of microcosm preparation on rates of toluene biodegradation under denitrifying conditions. J Ind Microbiol Biotech 18:170–176

    Google Scholar 

  • Illman W, Alvarez PJJ (2009) Performance assessment of bioremediation and natural attenuation. Crit Rev Environ Sci Technol 39:209–270

    Article  CAS  Google Scholar 

  • Jeffrey WH, Nazaret S, Barkay T (1996) Detection of the merA gene and its expression in the environment. Microb Ecol 32:293–303

    Article  CAS  PubMed  Google Scholar 

  • Kane SR, Beller HR, Legler TC, Anderson RT (2002) Biochemical and genetic evidence of benzylsuccinate synthase in toluene-degrading, ferric iron-reducing Geobacter metallireducens. Biodegradation 13:149–154

    Article  CAS  PubMed  Google Scholar 

  • Krajmalnik-Brown R, Holscher T, Thomson IN, Saunders MF, Ritalahti KM, Loffler FE (2004) Genetic identification of a putative vinyl chloride reductase in Dehalococcoides sp. strain BAV1. Appl Environ Microbiol 70:6347–6351

    Article  CAS  PubMed  Google Scholar 

  • Nakatsu CH, Carmosini N, Baldwin B, Beasly F, Kourtev P, Konopka A (2005) Soil microbial community responses to additions of organic carbon substrates and heavy metals (Pb and Cr). Appl Environ Microbiol 71:7679–7689

    Article  CAS  PubMed  Google Scholar 

  • Nazaret S, Jeffrey WH, Saouter E, Haven RV, Barkay T (1994) merA Gene expression in aquatic environments measured by mRNA production and Hg(II) volatilization. Appl Environ Microbiol 60:4059–4065

    CAS  PubMed  Google Scholar 

  • Neidhardt FC (1987) Chemical composition of Escherichia coli. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium: cellular and molecular biology, edn 1, vol 1. American Society for Microbiology, Washington, DC, pp 3–6

    Google Scholar 

  • Park JW, Crowley DE (2006) Dynamic changes in nahAc gene copy numbers during degradation of naphthalene in PAH-contaminated soils. Appl Microbiol Biotechnol 72:1322–1329

    Article  CAS  PubMed  Google Scholar 

  • Porteous LA, Seidler RJ, Watrud LS (1997) An improved method for purifying DNA from soil for polymerase chain reaction amplification and molecular ecology applications. Mol Ecol 6(8):787–791

    Article  CAS  Google Scholar 

  • Rabus R, Heider J (1998) Initial reactions of anaerobic metabolism of alkylbenzenes in denitrifying and sulfate-reducing bacteria. Arch Microbiol 170:377–384

    Article  CAS  Google Scholar 

  • Ritalahti KM, Amos KK, Sung Y, Wu Q, Koenigsberg SS, Löffler FE (2006) Quantitative PCR targeting 16S rRNA and reductive dehalogenase genes simultaneously monitors multiple Dehalococcoides strains. Appl Environ Microbiol 72:2765–2774

    Article  CAS  PubMed  Google Scholar 

  • Scow KM, Hicks KA (2005) Natural attenuation and enhanced bioremediation of organic contaminants in groundwater. Curr Opin Biotechnol 16:246–253

    Article  CAS  PubMed  Google Scholar 

  • Spormann AM, Widdel F (2000) Metabolism of alkylbenzenes, alkanes, and other hydrocarbons in anaerobic bacteria. Biodegradation 11:85–105

    Article  CAS  PubMed  Google Scholar 

  • Suzuki MT, Taylor LT, Delong EF (2000) Quantitative analysis of small-subunit rRNA genes in mixed microbial populations via 59-nuclease assays. Appl Environ Microbiol 66:4605–4614

    Article  CAS  PubMed  Google Scholar 

  • Von Gunten U, Zobrist J (1993) Biogeochemical changes in groundwater-infiltration systems: column studies. J Geochim Cosmochim Acta 57:3895–3906

    Article  Google Scholar 

  • Weiss JV, Cozzarelli IM (2008) Biodegradation in contaminated aquifers: incorporating microbial molecular methods. Ground Water 46:305–322

    Article  CAS  PubMed  Google Scholar 

  • Winderl C, Schaefer S, Lueders T (2007) Detection of anaerobic toluene and hydrocarbon degraders in contaminated aquifers using benzylsuccinate synthase (bssA) genes as a functional marker. Environ Microbiol 9:1035–1046

    Article  CAS  PubMed  Google Scholar 

  • Winderl C, Anneser B, Griebler C, Meckenstock RU, Lueders T (2008) Depth-resolved quantification of anaerobic toluene degraders and aquifer microbial community patterns in distinct redox zones of a tar oil contaminated plume. Appl Environ Microbiol 74:792–801

    Article  CAS  PubMed  Google Scholar 

  • Zengler K, Heider J, Rossello-Mora R, Widdel F (1999) Phototrophic utilization of toluene under anoxic conditions by a new strain of Blastochloris sulfoviridis. Arch Microbiol 172:204–212

    Article  CAS  PubMed  Google Scholar 

  • Zhou JZ, Bruns MA, Tiedje JM (1996) DNA recovery from soils of diverse composition. Appl Environ Microbiol 62(2):316–322

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Rick Ahlers for providing the aquifer material samples used in this work. Funding for this was provided by NSF (CBET-0729700). Amy Monier was partially supported by the American Petroleum Institute.

Author information

Author notes

  1. Sufia K. Kazy

    Present address: Department of Biotechnology, National Institute of Technology Durgapur, Mahatma Gandhi Avenue, Durgapur, WB, 713209, India

Authors and Affiliations

  1. Department of Civil and Environmental Engineering, Rice University, MS 317; 6100 Main Street, Houston, TX, 77005, USA

    Sufia K. Kazy, Amy L. Monier & Pedro J. J. Alvarez

Authors
  1. Sufia K. Kazy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amy L. Monier
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pedro J. J. Alvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pedro J. J. Alvarez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kazy, S.K., Monier, A.L. & Alvarez, P.J.J. Assessing the correlation between anaerobic toluene degradation activity and bssA concentrations in hydrocarbon-contaminated aquifer material. Biodegradation 21, 793–800 (2010). https://doi.org/10.1007/s10532-010-9344-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10532-010-9344-1

Keywords

Search

Navigation