Abstract
A mixed methane-oxidizing biofilm was characterized, concurrently using a number of advanced techniques. Community analysis results by microarray exhibited that type II members dominated the methanotrophic community, in which Methylocystis was most abundant, followed by Methylosinus. Observation results by fluorescent in situ hybridization and confocal microscopy showed multiple biofilm colonies that were irregular, bell-shaped, with mean thickness of approximately 20 μm. Image analysis results indicated that the relative abundance of methanotrophs peaked at a depth of about 5 μm. Although the biofilm colonies differed in size, methanotrophs accounted for 4–9%. Gaussian and linear regression results between the biofilm volumes and types I (r 2 = 0.86) and II volumes (r 2 = 0.92), respectively, revealed that type I members played a role in the growth of the biofilm but only below a threshold volume, whereas type II members supported the overall growth. Geostatistical analyses results revealed concentration of types I and II methanotrophic individuals with decreasing depth, and randomness between the spatial locations and population levels. Collectively, the methane-oxidizing biofilm was a highly organized system with methanotrophs and their cohabitants.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amaral JA, Archambault C, Richards SR, Knowles R (1995) Denitrification associated with groups I and II methanotrophs in a gradient enrichment system. FEMS Microbiol Ecol 18(4):289–298
Beyenal H, Lewandowski Z (2002) Internal and external mass transfer in biofilms grown at various flow velocities. Biotechnol Progr 18(1):55–61. doi:10.1021/bp010129s
Bodrossy L, Stralis-Pavese N, Murrell JC, Radajewski S, Weilharter A, Sessitsch A (2003) Development and validation of a diagnostic microbial microarray for methanotrophs. Environ Microbiol 5(7):566–582
Christensen BB, Sternberg C, Andersen JB, Eberl L, Moller S, Givskov M, Molin S (1998) Establishment of new genetic traits in a microbial biofilm community. Appl Environ Microbiol 64(6):2247–2255
Clapp LW, Regan JM, Ali F, Newman JD, Park JK, Noguera DR (1999) Activity, structure, and stratification of membrane attached methanotrophic biofilms cometabolically degrading trichloroethylene. Water Sci Technol 39(7):153–161
de Beer D, Stoodley P, Roe F, Lewandowski Z (1994) Effects of biofilm structures on oxygen distribution and mass transport. Biotchnol Bioeng 43(11):1131–1138. doi:10.1002/bit.260431118
Eller G, Stubner S, Frenzel P (2001) Group-specific 16S rRNA targeted probes for the detection of type I and type II methanotrophs by fluorescence in situ hybridisation. FEMS Microbiol Lett 198(2):91–97
Gebert J, Gröngröft A, Schloter M, Gattinger A (2004) Community structure in a methanotroph biofilter as revealed by phospholipid fatty acid analysis. FEMS Microbiol Lett 240(1):61–68
Gebert J, Stralis-Pavese N, Alawi M, Bodrossy L (2008) Analysis of methanotrophic communities in landfill biofilters using diagnostic microarray. Environ Microbiol 10(5):1175–1188
Graham DW, Chaudhary JA, Hanson RS, Arnold RG (1993) Factors affecting competition between type I and type II methanotrophs in two-organism, continuous-flow reactors. Microb Ecol 25(1):1–17
Hanson R, Hanson T (1996) Methanotrophic bacteria. Microbiol Rev 60(2):439–471
Henckel T, Roslev P, Conrad R (2000) Effects of O2 and CH4 on presence and activity of the indigenous methanotrophic community in rice field soil. Environ Microbiol 2(6):666–679
Lee E-H, Park H, Cho K-S (2010) Characterization of methane, benzene and toluene-oxidizing consortia enriched from landfill and riparian wetland soils. J Hazard Mater 184(1–3):313–320
Martineau C, Whyte LG, Greer CW (2010) Stable isotope probing analysis of the diversity and activity of methanotrophic bacteria in soils from the Canadian high arctic. Appl Environ Microbiol 76(17):5773–5784. doi:10.1128/aem.03094-09
McDonald IR, Upton M, Hall G, Pickup RW, Edwards C, Saunders JR, Ritchie DA, Murrell JC (1999) Molecular ecological analysis of methanogens and methanotrophs in blanket bog peat. Microb Ecol 38(3):225–233
Modin O, Fukushi K, Yamamoto K (2008) Simultaneous removal of nitrate and pesticides from groundwater using a methane-fed membrane biofilm reactor. Water Sci Technol 58(6):1273–1279
Modin O, Fukushi K, Nakajima F, Yamamoto K (2010) Nitrate removal and biofilm characteristics in methanotrophic membrane biofilm reactors with various gas supply regimes. Water Res 44(1):85–96
Murrell JC, Dalton H (1983) Nitrogen fixation in obligate methanotrophs. J Gen Microbiol 129(11):3481–3486. doi:10.1099/00221287-129-11-3481
Qureshi N, Annous B, Ezeji T, Karcher P, Maddox I (2005) Biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates. Microb Cell Fact 4(1):24
Rainey FA, Ward-Rainey NL, Janssen PH, Hippe H, Stackebrandt E (1996) Clostridium paradoxum DSM 7308T contains multiple 16S rRNA genes with heterogeneous intervening sequences. Microbiology 142(8):2087–2095. doi:10.1099/13500872-142-8-2087
Rittmann BE (2006) Microbial ecology to manage processes in environmental biotechnology. Trends Biotechnol 24(6):261–266
Scheutz C, Kjeldsen P, Bogner JE, De Visscher A, Gebert J, Hilger HA, Huber-Humer M, Spokas K (2009) Microbial methane oxidation processes and technologies for mitigation of landfill gas emissions. Waste Manag Res 27(5):409–455. doi:10.1177/0734242x09339325
Semrau JD, DiSpirito AA, Yoon S (2010) Methanotrophs and copper. FEMS Microbiol Rev 34(4):1–36
Stewart PS (2003) Diffusion in biofilms. J Bacteriol 185(5):1485–1491. doi:10.1128/jb.185.5.1485-1491.2003
Takeda K (1988) Characteristics of a nitrogen-fixing methanotroph, Methylocystis T-1. Antonie Leeuwenhoek 54(6):521–534. doi:10.1007/bf00588388
Tolker-Nielsen T, Molin S (2000) Spatial organization of microbial biofilm communities. Microb Ecol 40(2):75–84
Whittenbury R, Phillips KC, Wilkinson JF (1970) Enrichment, isolation and some properties of methane-utilizing bacteria. J Gen Microbiol 61(2):205–218. doi:10.1099/00221287-61-2-205
Yoon S, Carey J, Semrau J (2009) Feasibility of atmospheric methane removal using methanotrophic biotrickling filters. Appl Microbiol Biotechnol 83(5):949–956. doi:10.1007/s00253-009-1977-9
Acknowledgments
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRL program, R0A-2008-000-20044-0), and RP-Grant 2011 of Ewha Womans University.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(DOCX 2670 kb)
Rights and permissions
About this article
Cite this article
Kim, T.G., Yi, T., Lee, EH. et al. Characterization of a methane-oxidizing biofilm using microarray, and confocal microscopy with image and geostatic analyses. Appl Microbiol Biotechnol 95, 1051–1059 (2012). https://doi.org/10.1007/s00253-011-3728-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-011-3728-y