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Bacterial community dynamics during in-situ bioremediation of petroleum waste sludge in landfarming sites

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Abstract

In-situ bioremediation of petroleum waste sludge in landfarming sites of Motor Oil Hellas (petroleum refinery) was studied by monitoring the changes of the petroleum composition of the waste sludge, as well as the changes in the structure of the microbial community, for a time period of 14 months. The analyses indicated an enhanced degradation of the petroleum hydrocarbons in the landfarming areas. A depletion of n-alkanes of approximately 75–100% was obtained. Marked changes of the microbial communities of the landfarms occurred concomitantly with the degradation of the petroleum hydrocarbons. The results obtained from terminal restriction fragment length polymorphism (T-RFLP) analysis of polymerase chain reaction (PCR) amplified 16S rRNA genes demonstrated that bacteria originating from the refinery waste sludge and newly selected bacteria dominated the soil bacterial community during the period of the highest degradation activity. However, the diversity of the microbial community was decreased with increased degradation of the petroleum hydrocarbons contained in the landfarms. T-RFLP fingerprints of bacteria of the genera Enterobacter and Ochrobactrum were detected in the landfarmed soil over the entire treatment period of 14 months. In contrast, the genus Alcaligenes appeared in significant numbers only within the 10 month old landfarmed soil. Genes encoding catechol 2,3-dioxygenase (subfamily I.2.A) were detected only in DNA of the untreated refinery waste sludge. However, none of the genes known to encode the enzymes alkane hydroxylase AlkB, catechol 2,3-dioxygenase (subfamily I.2.A) and naphthalene dioxygenase nahAc could be detected in DNA of the landfarmed soils.

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Abbreviations

OTUs:

Operational Taxonomic Units

PCR:

Polymerase chain reaction

T-RFLP:

Terminal restriction fragment length polymorphism fingerprinting

References

  • RI Amann W Ludwig K-H Schleifer (1995) ArticleTitlePhylogenetic identification and in situ detection of individual microbial cells without cultivation Microbiol. Rev. 59 143–169 Occurrence Handle1:CAS:528:DyaK2MXkvVGmurk%3D

    CAS  Google Scholar 

  • J Borneman PW Skroch KM O’Sullivan JA Palus NG Rumjanek JL Jansen J Nienhuis E Triplett (1996) ArticleTitleMolecular microbial diversity of an agricultural soil in Wisconsin Appl. Environ. Microbiol. 62 IssueID6 1935–1943 Occurrence Handle1:CAS:528:DyaK28XjtlGgsr0%3D

    CAS  Google Scholar 

  • TD Brock (1987) ArticleTitleThe study of microorganisms in situ: progress and problems Symp. Soc. Gen. Microbiol. 41 1–17

    Google Scholar 

  • A Felske H Rheims A Wolterink E Stackebrandt ADL Akkermanns (1997) ArticleTitleRibosome analysis reveals prominent activity of an uncultured member of the class Actinobacteria in grassland soils Microbiology 143 2983–2989 Occurrence Handle10.1099/00221287-143-9-2983 Occurrence Handle1:CAS:528:DyaK2sXmt1yksL0%3D

    Article  CAS  Google Scholar 

  • N Hamamura CM Yeager DJ Arp (2001) ArticleTitleTwo distinct monooxygenases for alkane oxidation in Nocardioides sp strain CF8 Appl. Environ. Microbiol. 67 IssueID11 4992–4998 Occurrence Handle1:CAS:528:DC%2BD3MXotlSgurs%3D

    CAS  Google Scholar 

  • I Head JR Saunders RW Pickup (1998) ArticleTitleMicrobial evolution, diversity, and ecology: a decade of ribosomal RNA analysis of uncultivated microorganisms Microbiol. Ecol. 35 1–21 Occurrence Handle1:CAS:528:DyaK1cXks1SitA%3D%3D

    CAS  Google Scholar 

  • BP Hedlung AD Geiselbrecht TJ Bair JT Staley (1999) ArticleTitlePolycyclic aromatic hydrocarbon degradation by a new marine bacterium, Neptunomonas naphthovorans gen nov., sp. nov. Appl. Environ. Microbiol. 65 IssueID1 251–259

    Google Scholar 

  • H Heuer M Krsek P Baker K Smalla EMH Wellington (1997) ArticleTitleAnalysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoresis separation in denaturing gradients Appl. Environ. Microbiol. 63 3233–3241 Occurrence Handle1:CAS:528:DyaK2sXlt1Wis7c%3D

    CAS  Google Scholar 

  • CW Kaplan CL Kitts (2004) ArticleTitleBacterial succession in a petroleum land treatment unit Appl. Environ. Microbiol. 70 IssueID3 1777–1786 Occurrence Handle1:CAS:528:DC%2BD2cXisVKisLc%3D

    CAS  Google Scholar 

  • E Katsivela ERB Moore N Kalogerakis (2003a) ArticleTitleBiodegradation of aliphatic and aromatic hydrocarbons: specificity among bacteria isolated from refinery waste sludge Water, Air Soil Poll.: Focus 3 103–115 Occurrence Handle1:CAS:528:DC%2BD3sXktFGrsLg%3D

    CAS  Google Scholar 

  • Katsivela E, Maroukli D, Moore ERB, Weller R, Heuer H, Pieper D & Kalogerakis N (2003b) Degradation of petroleum hydrocarbons by a newly isolated Alcaligenes species and monitoring of its relative abundance in landfarming sites. In: Proceedings of the Second European Bioremediation Conference, Chania, Greece, June 30--July 4, 2003, pp. 416--419

  • E Katsivela ERB Moore N Kalogerakis (2004) ArticleTitleMonitoring of the degradation activities and the diversity of the microbial community degrading petroleum waste sludge Water, Air Soil Poll.: Focus 4 75–85 Occurrence Handle1:CAS:528:DC%2BD2cXosVyjtL0%3D

    CAS  Google Scholar 

  • D-H Lee Y-G Zo S-J Kim (1996) ArticleTitleNonradioactive method to study genetic profiles of natural bacterial communities by PCR-single-strand-conformation polymorphism Appl. Environ. Microbiol. 62 3112–3120 Occurrence Handle1:CAS:528:DyaK28XlsFert70%3D

    CAS  Google Scholar 

  • WT Liu TL Marsh H Cheng LJ Forney (1997) ArticleTitleCharacterization of microbial diversity by determining terminal restriction fragment length polymorphism of genes encoding 16S rRNA Appl. Environ. Microbiol. 63 4516–4522 Occurrence Handle1:CAS:528:DyaK2sXnt12ntbs%3D

    CAS  Google Scholar 

  • BM Mesarch HC Nakatsu L Nies (2000) ArticleTitleDevelopment of catechol 2,3-dioxygenase-specific primers for monitoring bioremediation by competitive quantitative PCR Appl. Environ. Microbiol. 66 IssueID2 678–683 Occurrence Handle1:CAS:528:DC%2BD3cXhtFeqsrY%3D

    CAS  Google Scholar 

  • Moore ERB, Arnscheidt A, Krüger A, Strömpl C, Mau M (1999) Simplified protocols for the preparation of genomic DNA from bacterial cultures. In: Akkermans ADL, van Elsas JD & de Bruijn FJ (Eds) Molecular Microbial Ecology Manual, Vol 1.6.1 (pp 1--15). Kluwer Academic Press

  • ERB Moore M Mau A Arnscheidt EC Böttger RA Hutson MD Collins Y Peer ParticleVan De R Wachter ParticleDe KN Timmis (1996) ArticleTitleThe determination and comparison of the 16S rRNA gene sequences of species of the genus Pseudomonas (sensu stricto) and estimation of the natural intrageneric relationships System. Appl. Microbiol. 19 476–492

    Google Scholar 

  • G Muyzer EC Waal ParticleDe AG Uitterlinden (1993) ArticleTitleProfiling of complex microbial populations by denaturing gradient gel electrophoresis analysis by polymerase chain reaction-amplified genes coding for 16S rRNA Appl. Environ. Microbiol. 59 695–700 Occurrence Handle1:CAS:528:DyaK3sXit1Kktrk%3D

    CAS  Google Scholar 

  • B Nogales ERB Moore E Llobet-Brossa R Rosselo-Mora R Amann KN Timmis (2001) ArticleTitleCombined use of 16S ribosomal DANN and 16S rRNA to study the bacterial community of polychlorinated biphenyl-polluted soil Appl. Environ. Microbiol. 67 IssueID4 1874–1884 Occurrence Handle1:CAS:528:DC%2BD3MXis1egtbk%3D

    CAS  Google Scholar 

  • MA Osborn ERB Moore KN Timmis (2000) ArticleTitleAn evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics Environ. Microbiol. 2 IssueID1 39–50 Occurrence Handle1:CAS:528:DC%2BD3cXhs12ht7k%3D

    CAS  Google Scholar 

  • NR Pace (1997) ArticleTitleA molecular view of microbial diversity and the biosphere Science 276 734–740 Occurrence Handle1:CAS:528:DyaK2sXivFOltLc%3D

    CAS  Google Scholar 

  • F Schwieger CC Tebbe (1998) ArticleTitleA new approach to utilize PCR-single-strand-conformation polymorphism for 16S rRNA gene-based microbial community analysis Appl. Environ. Microbiol. 64 4870–4876 Occurrence Handle1:CAS:528:DyaK1cXnvF2htrs%3D

    CAS  Google Scholar 

  • MJ Simon TD Osslund R Saunders BD Ensley S Suggs A Harcourt W-C Suen DL Cruden DT Gibson GJ Zylstra (1993) ArticleTitleSequences of genes encoding naphthalene dioxygenase in Pseudomonas putida strains G7 and NCIB 9816-4 Gene 127 31–37 Occurrence Handle1:CAS:528:DyaK3sXks1Cls7g%3D

    CAS  Google Scholar 

  • THM Smits M Röthlisberger B Witholt JB Beilen Particlevan (1999) ArticleTitleMolecular screening for alkane hydroxylase genes in gram-negative and gram-positive strains Environ. Microbiol. 1 IssueID4 307–317 Occurrence Handle1:CAS:528:DyaK1MXmtFOrtbk%3D

    CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Biochemical Engineering & Environmental Biotechnology, Department of Environmental Engineering, Technical University of Crete, Polytechneioupolis, Chania, GR-73100, Greece

    E. Katsivela, D. Maroukli & N. Kalogerakis

  2. Department of Natural Resources and Environment, Technological Educational Institute of Crete, Romanou 3, Halepa, GR- 73133, Chania, Crete, Greece

    E. Katsivela & D. Maroukli

  3. Macaulay Institute, AB15 8QH, Craigiebuckler, Aberdeen, Scotland, UK

    E. R. B. Moore

  4. Division of Microbiology, National Research Centre for Biotechnology (GBF), Mascheroder Weg 1, D-38124, Braunschweig, Germany

    C. Strömpl & D. Pieper

Authors
  1. E. Katsivela
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  2. E. R. B. Moore
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  3. D. Maroukli
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  4. C. Strömpl
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  5. D. Pieper
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  6. N. Kalogerakis
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Correspondence to E. Katsivela.

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Katsivela, E., Moore, E.R.B., Maroukli, D. et al. Bacterial community dynamics during in-situ bioremediation of petroleum waste sludge in landfarming sites. Biodegradation 16, 169–180 (2005). https://doi.org/10.1007/s10532-004-4883-y

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  • DOI: https://doi.org/10.1007/s10532-004-4883-y

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