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Use of Microautoradiography to Study in situ Physiology of Bacteria in Biofilms

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Abstract

Microautoradiography is a technique that enables direct detection of active bacteria in complex microbial systems on a single cell level. When combined with fluorescence in situ hybridization using oligonucleotide probes for identification of the microorganisms, it is possible to link key physiological features to the identity of microorganism. In this way information can be obtained about the physiology of cultured as well as not-yet cultured microorganism in biofilms and other complex ecosystems in a way that presently cannot be provided by other methods. In this small review we describe the basic procedure, the type of information that can be obtained, the most important limitations by using this approach, and some recent examples from biofilms and other complex microbial communities.

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References

  • Andreasen K & Nielsen PH (1997)Application of microautoradiography for the study of substrate uptake by lamentous microorganisms in activated sludge.Appl.Environ.Microbiol.63:3662–3668

    Google Scholar 

  • Andreasen K & Nielsen PH (2000)Growth of Microthrix parvicella in nutrient removal activated sludge plants:Studies of in situ physiology.Water Res.34:1559–1569

    Google Scholar 

  • Bourdier G, Bohatier M & Feuillade J (1989) Amino acid incorporation by natural population of Oscillatoria rubescens. A microautoradiographic study. FEMS Microbiol. Ecol.62:185–190

    Google Scholar 

  • Brock TD & Brock ML (1966)Autoradiography as a tool in microbial ecology.Nature 209:734–736

    Google Scholar 

  • Brock ML & Brock TD (1968)The application of microautoradiographic techniques to microbial ecology.Fur Theor. Ang.Limnol.15:1–29

    Google Scholar 

  • Carman KR (1990)Radioactive labelling of a natural assemblage of marine sedimentary bacteria and microalgae for the trophic studies:An autoradiographic study.Microbial Ecol. 19:279–290

    Google Scholar 

  • Carman K (1993)Microautoradiographic detection of microbial activity.In:Kemp PF, Sherr BF, Sherr EB & Cole JJ (eds)Handbook of Methods in Aquatic Microbial Ecology (pp 397–404).Lewis Publishers, London

    Google Scholar 

  • Cottrell MT & Kirchman DL (2000)Natural assemblage of marine proteobacteria and members of the Cytophaga.avobacter cluster consuming low and high-molecularweight dissolved organic matter.Appl.Enivron.Microbiol.66: 1360–1363

    Google Scholar 

  • Daims H, Bruehl A, Amann R, Schleifer KH & Wagner M (1999)The Domain-speci c probe EUB338 is insufficient for the detection of all Bacteria:Development and evaluation of a more comprehensive probe set.Syst.Appl.Microbiol.22: 434–444

    Google Scholar 

  • Daims H, Nielsen JL, Nielsen PH, Schleifer K-H & Wagner M (2001)In situ characterization of Nitrospira-like nitrite-oxidizing bacteria active in wastewater treatment plants.Appl.Environ.Microbiol.67:5273–5284

    Google Scholar 

  • Fliermans CB & Schmidt EL (1975)Autoradiography and immuno-.uorescence combined for autecological study of single cell activity with Nitrobacter as a model system.Appl. Microbiol.30:676–684

    Google Scholar 

  • Gray ND, Howarth R, Pickup RW, Jones JG & Head IM (2000) Use of combined microautoradiography and fluorescence in situ hybridization to determine carbon metabolism in mixed natural communities of uncultured bacteria from the genus Achromatium. Appl.Environ.Microbiol.66:4518–4522

    Google Scholar 

  • Grossmann S (1994)Bacterial activity in sea ice and open water of the Weddel Sea,Antarctica:A microautoradiographic study.Microbial.Ecol.28:1–18

    Google Scholar 

  • Hermansson M & Dahlbäck B (1983)Bacterial activity at the air/water interface.Microbial Ecol.9:317–328

    Google Scholar 

  • Hoppe H-G (1976)Determination and properties of actively metabolizing heterotrophic bacteria in the sea,investigated by means of micro-autoradiography.Mar.Bio.36:291–302

    Google Scholar 

  • Ito T, Nielsen JL, Okabe S, Watanabe Y & Nielsen PH (2002) Phylogenetic identi cation and substrate uptake patterns of sulfate-reducing bacteria inhabiting an aerobic–anaerobic sewer bio lm.Appl.Environ.Microbiol.68:356–364

    Google Scholar 

  • Lee N, Nielsen PH, Andreasen K, Juretschko S, Nielsen JL, Schleifer K-H & Wagner M (1999)Combination offluorescent in situ hybridization and microautoradiography–a new tool for structure-function analysis in microbial ecology. Appl.Environ.Microbiol.65:1289–1297

    Google Scholar 

  • Lee N, Nielsen PH, Aspegren H, Henze M, Schleifer K-H & la Cour Jansen J (2003)Long-term population dynamics and in situ physiology in activated sludge systems with enhanced biological phosphorus removal operated with and without nitrogen removal.Sys.Appl.Microbiol.26:211–227

    Google Scholar 

  • Meyer-Reil L-A (1978)Autoradiography and epifluorescence microscopy combined for the determination of number and spectrum of actively metabolizing bacteria in natural waters. Appl.Environ.Microbiol.36:506–512

    Google Scholar 

  • Nielsen PH, Andreasen K, Lee N & Wagner M (1999)Use of microautoradiography and fluorescent in situ hybridization for characterization of microbial activity in activated sludge. Water Sci.Technol.39(1):1–9

    Google Scholar 

  • Nielsen PH Aquino de Muro M & Nielsen JL (2000)Studies on the in situ physiology of Thiothrix spp.in activated sludge. Environ.Microbiol.2:389–398

    Google Scholar 

  • Nielsen JL, Christensen D, Kloppenborg M & Nielsen PH (2003)Quanti cation of cell-speci c substrate uptake by probe-de ned bacteria under in situ conditions by microautoradiography and fluorescence in situ hybridization.Environ.Microbiol.5(3):202–211

    Google Scholar 

  • Nielsen JL & Nielsen PH (2002a)Enumeration of acetateconsuming bacteria by microautoradiography under oxygenand nitrate respiring conditions in activated sludge.Water Res.36:421–428

    Google Scholar 

  • Nielsen JL & Nielsen PH (2002b)Enumeration of functional bacterial groups in activated sludge by microautoradiography.Water Sci.Tech.46(1–2):389–395

    Google Scholar 

  • Nielsen PH, Roslev P, Dueholm T & Nielsen JL (2002) Microthrix parvicella, a specialized lipid consumer in anaerobic–aerobic activated sludge plants Water Sci.Technol. 46(1–2):73–80

    Google Scholar 

  • Nilsson C & Sundbäck K (1996)Amino acid uptake in natural microphytobenthic assemblages studied by microautoradiography.Hydrobiologia 332:119–129

    Google Scholar 

  • Ouverney CC & Fuhrman JA (1999)Combined microautoradiography-16S rRNA probe technique for determination of radioisotope uptake by speci c microbial cell type in situ. Appl.Enivron.Microbiol.65:1746–1752

    Google Scholar 

  • Ouverney CC & Fuhrman JA (2000)Marine planktonic Archaea take up amino acids.Appl.Enivron.Microbiol. 66:4829–4833

    Google Scholar 

  • Pearl HW (1984)Alteration of metabolic activities in association with detritus.Bull.Mar.Sci.35:393–408

    Google Scholar 

  • Pearl HW (1991)Ecophysiological and tropical implications of light-stimulated amino acid utilization in marine picoplankton.Appl.Environ.Microbiol.57:473–479

    Google Scholar 

  • Pearl HW & Priscu JC (1998)Microbial phototrophic,heterotrophic and diazotrophic activities associated with aggregates in the permanent ice cover of lake Bonney,Antarctica. Microbial Ecol.36:221–230

    Google Scholar 

  • Pearl HW & Stull EA (1979)In defense of grain density autoradiography.Limnol.Oceanogr.24:1166–1169

    Google Scholar 

  • Pedrós-Alió C & Nedwell S (1989)Microautoradiographic study of thymidine uptake in brackish waters around Sapelo Island,Georgia,USA.Mar.Ecol-Prog.Ser.55:83–94

    Google Scholar 

  • Rogers AW (1979)Techniques of Autoradiography.Elsevier, New York

    Google Scholar 

  • Roszak DB & Colwell RR (1987)Metabolic activity of bacteria cells enumerated by direct viable count.Appl.Environ. Microbiol.53:2889–2983

    Google Scholar 

  • Tabor SP & Neihof RA (1982)Improved microautoradiographic method to determine individual microorganisms active in substrate uptake in natural waters.Appl.Environ. Microbiol.44:945–953

    Google Scholar 

  • Tabor SP & Neihof RA (1984)Direct determination of activities for microorganisms of Chesapeake Bay populations.Appl.Environ.Microbiol.48:1012–1019

    Google Scholar 

  • Thomsen TR, Kjellerup BV, Nielsen JL, Hugenholtz P & Nielsen PH (2002)In situ studies of the phylogeny and physiology of lamentous bacteria with attached growth. Environ.Microbiol.4:383–391

    Google Scholar 

  • Vaniman DA, Furlano S, Chipera J, Thompson K & Triay I (1996)Microautoradiography in studies of Pu(V)sorption by trace and fracture minerals in tuff.Mat.Res.Soc.Symp. Proc.412:639–646

    Google Scholar 

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Nielsen, J.L., Wagner, M. & Nielsen, P.H. Use of Microautoradiography to Study in situ Physiology of Bacteria in Biofilms. Re/Views in Environmental Science and Bio/Technology 2, 261–268 (2003). https://doi.org/10.1023/B:RESB.0000040464.52512.44

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  • DOI: https://doi.org/10.1023/B:RESB.0000040464.52512.44

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