enzymatic, catalytic and chemical routes. A possible route bilization by sulfate-reducing biofilms. Environ. Sci.

for uranium reduction in anoxic conditions leading to pre- Technol.

cipitated uraninite UO2(s), which is likely to happen in the 38, 2067–2074.
multispecies biofilms under investigation is the reduction Bishop P. L. and Yu T. (1999) A microelectrode study of
redox potential change in biofilms. Water Sci. Technol.
of uranyl(VI) by EPS components, e.g., glucose and the
39(7), 179–
simultaneous oxidation of glucose to gluconic acid: 185.
Bouwer E. J., Rijnaarts H. H. M., Cunningham A. B. and Gerlach
R. (2000) Biofilms in porous media. In Biofilms II: Analysis,
C6 H12 O6 þ UO22þ þ H2 O ¼ C6 H12 O7 þ UO2ðsÞ þ Processes, and Applications (ed. J. D. Bryers). Wiley-Liss, New
York, pp. 123–158.
2Hþ
Under oxic conditions strong oxidizing agents, e.g., chro- Beyenal H., Sani R. K., Peyton B. M., Dohnalkova
mium trioxide (Herscovici et al., 1982) are needed to A. C., Amonette J. E. and Lewandowski Z. (2004)
oxidize glucose or other EPS components. However, under Uranium immo-
anoxic conditions the above specified reaction proceeds
without such strong oxidizing agents and UO2 2þ may act as
electron
acceptor leading to precipitated uraninite UO2(s). Since the
increased respiratory activity, as a result of the metabolic
response to the added uranium, induced larger O2 depleted
zones within the biofilms, it will be much easier to immobi-
lize uranium in such zones as precipitated uranium. Conse-
quently, such expanded anoxic zones may be responsible
for an increased immobilization of uranium from the sur-
rounding bulk solution. Our studies indicated that the
microbial influence on the migration behavior of uranium
in the environment has to be included in performance
assessment studies to predict more realistic uranium migra-
tion scenarios.

ACKNOWLEDGMENTS

The authors thank Peter Berg for valuable discussions concern-

ing the numerical model PROFILE, Katrin Flemming for 16S
rDNA analysis. Roy Wogelius and two anonymous reviewer are
thanked for constructive suggestions helping to improve the man-
uscript. In addition, funding by the German Research Council
(DFG) under the Contract No. AR 584/1-1 and by the EU within
the FUNMIG project is gratefully acknowledged.

REFERENCES

Abdelouas A., Lu Y., Lutze W. and Nuttall H. E. (1998)

Reduction of U VI to U IV by indigenous bacteria in
contaminated ground water. J. Contam. Hydrol. 35, 217–
233.
Amann R. and Ku¨ hl M. (1998) In situ methods for
assessment of microorganisms and their activities. Curr.
Opin. Microbiol. 1,
352–358.
Badireddy A. R., Chellam S., Yanina S., Gassmann P. and
Rosso K. M. (2008) Bismuth dimercaptopropanol
(BisBAL) inhibits the expression of extracellular
polysaccharides and proteins by Brevundimonas diminuta:
implications for membrane microfil- tration. Biotechnol.
Bioeng. 99(3), 634–643.
Barton L. L., Choudhury K., Thomson B. M., Steenhoudt
K. and Groffman A. R. (1996) Bacterial reduction of
soluble uranium: the first step of in situ immobilizati on
of uranium. Radioact. Waste Manag. Environ. Restor. 20,
141–151.
Berg P., Risgaard-Petersen N. and Rysgaard S. (1998) Interpreta-
tion of measured concentration profiles in sediment pore water.
Limnol. Oceanogr. 43(7), 1500–1510.
Bernhard G., Geipel G., Brendler V. and Nitsche H.
(1996) Speciation of uranium in seepage waters of a mine
tailing pile studied by time-resolved laser-induced
fluorescence spectros- copy (TRLFS). Radiochim. Acta 74,
87–91.
Broecker W. S. and Peng T.-H. (1974) Gas exchange rates
between air and sea. Tellus 26, 21–35.
Bryers J. D. (2000) Process engineering. In Biofilms II: Analysis,
Processes, and Applications (ed. J. D. Bryers). Wiley-Liss, New
York, pp. 13–44.
Chandler D. P., Fredrickson J. K. and Brockman F. J. (1997)
Effect of PCR template concentration on the
composition and distribution of total community 16S
rDNA clone libraries. Mol. Ecol. 6(5), 475–482.
Costerton J. W., Cheng K.-J., Geesey G. G., Ladd T. I., Nickel J.
C., Dasgupta M. and Marrie T. J. (1987). Annu. Rev. Microbiol.
41, 435–464.
Costerton J. W., Lewandowski Z., Caldwell D. E., Korber D. R.
and Lappin-Scott H. M. (1995) Microbial biofilms. Annu. Rev.
Microbiol. 49, 711–745.
de Beer (2000) Potentiometric microsensors for in situ measure-
ments in aquatic environments. In In Situ Monitoring of Aquatic
Systems (eds. J. Buffle and G. Horvai). Wiley, Chichester, pp.
161–194.
de Beer D. A. and Ku¨ hl M. (2001) Interfacial processes
and activities in biofilms and microbial mats. In The
Benthic Boundary Layer (eds. B. P. Boudreau and B. B.
Jørgensen). Oxford University Press, Oxford, pp. 374–394.
de Beer D., Stoodley P., Roe F. and Lewandowski Z. (1994)
Effects of biofilm structures on oxygen distribution and
mass transport. Biotechnol. Bioeng. 43, 1131–1138.
de Beer D., Schramm A., Santegoeds C. M. and Ku¨ hl M. (1997)
A nitrite microsensor for profiling environmental biofilms.
Appl. Environ. Microbiol. 63, 973–977.
de Wit D. (1992) Oxidation of Sucrose and Glucose to
Chiral Building Blocks. Ph.D. Thesis, Techn. Univ. Delft,
Nederlands, pp. 143.
Flemming H.-C. (1991) Biofilme und Wassertechnologie. Teil 1:
Entstehung, Aufbau, Zusammensetzung und Eigenschaften von
Biofilmen.. gwf-Wasser/Abwasser 132(4), 197–207.
Flemming H.-C., Leis A. and Wingender J. (2004) Biofilms and the
role of extracellular polymeric substances. In Advances in Biofilm
Science and Engineering (eds. R. Jordan, D. Williams and U.
Charaf). Cytergy Publishing, Bozeman, MT, pp. 86–142.
Francis A. J., Dodge C. J., Lu F., Halada G. P. and
Clayton C. R.
(1994) XPS and XANES studies of uranium
reduction by
Clostridium sp. Environ. Sci. Technol. 26, 636–639.
Fredrickson J. K., Zachara J. M., Kennedy D. W., Duff
M. C., Gorby Y. A., Li S. W. and Krupka K. M. (2000)
Reduction of U(VI) in goethite (R-FeOOH) suspensions
by a dissimilatory metalreducing bacterium. Geochim.
Cosmochim. Acta 64, 3085–
3098.
Ganesh R., Robinson K. G., Reed G. D. and Sayler G. S.
(1997) Reduction of hexavalent uranium from organic
complex by sulfate- and iron-reducing bacteria. Appl.
Environ. Microbiol.
63, 4385–4391.
Ganesh R., Robinson K. G., Chu L., Kucsmas D. and Reed
G. D. (1999) Reductive precipitation of uranium by
Desulfovibrio desulfuricans: evaluation of cocontaminant
effects and selective removal. Water Res. 33, 3447–3458.
Geipel G., Thieme M., Bernhard G. and Nitsche H. (1994)
Distribution of uranium and radionuclides in a uranium-mining
 Rockpile in Schlema, Saxony, Germany. Radiochim. Acta(66/ 2620–2624.
67), 305–308. Lloyd J. R. and Macaskie L. E. (2000) Bioremediation of
Geissler A. and Selenska-Pobell S. (2005) Addition of U(VI) radionuclide containing wastewaters. In Environmental
to a uranium mining waste sample and resulting Microbe–Metal Interactions (ed. D. R. Lovley). ASM Press,
changes in the indigenous bacterial community. Geobiology Washington, DC, pp. 277–327.
3(4), 275–285. Lovley D. R. (1995) Microbial reduction of iron,
Geslin C., Llanos J., Prieur D. and Jeanthon C. (2001) manganese, and other metals. In Advances in
The manganese and iron superoxide dismutases protect Agronomy, vol. 54 (eds. L.
Escherichia coli from heavy metal toxicity. Res. Microbiol.
152(10), 901–
905.
Gieseke A. and de Beer D. (2004) Use of
microelectrodes to measure in situ microbial activities in
biofilms, sediments, and microbial mats. In Molecular
Microbial Ecology Manual, second ed. 8.02 (eds. A. D.
L. Akkermans and D. van Elsas). Kluwer Academic
Publishers, Netherlands, pp. 1581–1612.
Grossmann K., Arnold T., Krawczyk-Ba¨ rsch E.,
Diessner S., Wobus A., Bernhard G. and Krawietz R.
(2007) Identification of fluorescent U(V) and U(VI)
microparticles in a multispecies biofilm by confocal laser
scanning microscopy and fluorescence spectroscopy. Environ.
Sci. Technol. 41, 6498–6504.
Hall-Stoodley L., Costerton J. W. and Stoodley P. (2004)
Bacterial biofilms: from the natural environment to
infectious diseases. Nat. Rev. Microbiol. 2, 95–108.
Harrison J. J., Ceri H. and Turner R. J. (2007)
Multimetal
resistance and tolerance in microbial biofilms. Nat.
Rev. Microbiol. 5, 928–938.
Henningsen D. and Katzung G. (2002) Einfu¨ hrung in die
Geologie Deutschlands. Spektrum, Akad. Verl., Heidelberg, pp.
214. Herscovici J., Egron M. J. and Antonakis K. (1982)
New Oxidative
systems for alcohols—molecular-sieves with chromium-
(VI)
reagents. J. Chem. Soc .Perkin Trans. 1(9), 1967–1973.
Holmes D. E., Finneran K. T., O’Neil R. A. and Lovley
D. R. (2002) Enrichment of members of the family
Geobacteraceae associated with stimulation of dissimilatory
metal reduction in uranium contaminated aquifer
sediments. Appl. Environ. Microbiol. 68, 2300–2306.
Hua B., Xu H., Terry J. and Deng B. (2006)
Kinetics of uranium(VI) reduction by hydrogen sulfide in
anoxic aqueous systems. Environ. Sci. Technol. 40, 4666–
4671.
Ji G. Y. and Silver S. (1995) Bacterial resistance
mechanisms for heavy metals of environmental concern. J.
Ind. Microbiol. 14(2),
61–75.
Kelly S. D., Kemner K. M., Carley J., Criddle C.,
Jardine P. M., Marsh T. L., Phillips D., Watson
D. and Wu W.-M. (2008) Speciation of uranium in
sediments before and after in situ biostimulation.
Environ. Sci. Technol. 42,
1558–1564.
Ku¨ hl M. (2005) Optical microsensors for analysis of
microbial communities. In Environmental Microbiology, vol.
397 (ed. J. R. Leadbetter). Methods in Enzymology, pp.
166–199.
Lane D. J. (1991) 16S/23S rRNA sequencing. In Nucleic Acid
Techniques in Bacterial Systematics (eds. E. Stackebrandt and
M. Goodfellow). John Wiley, Chichester, pp. 115–176.
Lawrence J. R., Korber D. R., Hoyle B. D., Costerton
J. W. and
Caldwell D. E. (1991) Optical sectioning of microbial
biofilms.
J. Bacteriol. 173, 6558–6567.
Lawrence J. R., Swerhone G. D. W. and Neu T. R. (2000) A
simple rotating annular reactor for replicated
biofilm studies. J. Microbiol. Methods 42, 215–224.
Lewandoski Z. (2000) Notes on biofilm porosity. Water Res. 34(9),
 Donald and L. Sparks). Academic Press, New York, pp. 175– Pons M. P. and Fuste´ M. C. (1993) Uranium
231. uptake by
Lovley D. R. and Phillips E. J. P. (1986) Organic immobilized cells of Pseudomonas strain EPS 5028.
matter mineralization with reduction of ferric iron Appl. Microbiol. Biotechnol. 39, 661–665.
in anaerobic sediments. Appl. Environ. Microbiol. 51, 683– Raffi J. and Selenska-Pobell S. (2006) Toxic avengers. Nucl. Eng.
689. Int. 51(619), 34–36.
Lovley D. R. and Phillips E. J. P. (1988) Novel mode of Revsbech N. P. (1989) An oxygen microsensor with
microbial energy metabolism: organic carbon oxidation a guard cathode. Limnol. Oceanorg. 34(2), 474–478.
coupled to dissimilatory reduction of iron or manganese.
Appl. Environ. Microbiol. 54, 1472–1480.
Lovley D. R. and Phillips E. J. P. (1992) Reduction of
uranium by
Desulfovibrio desulfuricans. Appl. Environ. Microbiol. 58,
850–
856.
Lovley D. R., Phillips E. J. P., Gorby Y. and Landa E. R.
(1991) Microbial reduction of uranium. Nature
350,
413–415.
Lovley D. R., Giovannoni S. J., White D. C., Champine J. E.,
Phillips E. J., Gorby Y. A. and Goodwin S. (1993a) Geobacter
metallireducens gen. nov. sp. nov., a microorganism capable of
coupling the complete oxidation of organic compounds to the
reduction of iron and other metals. Arch. Microbiol. 159(4),
336–344.
Lovley D. R., Roden E. E., Phillips E. J. P. and
Woodward J. C. (1993b) Enzymatic iron and uranium
reduction by sulfate- reducing bacteria. Mar. Geol. 113,
41–53.
Loy A., Maixner F., Wagner M. and Horn M.
(2007) probeBase—an online resource for rRNA-
targeted oligo- nucleotide probes: new features 2007.
Nucleic Acids Res. 35,
D800–D804.
Macaskie L. E., Bonthrone K. M., Yong P. and Goddard
D. T. (2000) Enzymically mediated bioprecipitation of
uranium by a Citrobacter sp.: a concerted role for
exocellular lipopolysac- charide and associated
phosphatase in biomineral formation. Microbiology 146,
1855–1867.
Manz W., Amann R. I., Ludwig W., Wagner M. and
Schleifer K.-
H. (1992) Phylogenetic oligodeoxynucleotide probes for
the major subclasses of Proteobacteria: problems and
solutions. Syst. Appl. Microbiol. 15, 593–600.
Merroun M. L., Raff J., Rossberg A., Hennig C.,
Reich T. and
Selenska-Pobell S. (2005) Complexation of uranium by
cells and S-layer sheets of Bacillus sphaericus JG-A12. App.
Environ. Microbiol. 71(9), 5532–5543.
Nakano S., Takeshita A., Ohtsuka T. and Nakai D. (2006)
Vertical
profiles of current velocity and dissolved oxygen
saturation in biofilms on artificial and natural substrates.
Limnology 7(3),
213–218.
Nies D. H. (2003) Efflux-mediated heavy metal
resistance in prokaryotes. FEMS Microbiol. 27(2-3), 313–
339.
Nyman J. L., Marsh T. L., Ginder-Vogel M. A.,
Gentile M.,
Fendof S. and Criddle C. S. (2005) Heterogeneous
response to biostimulation for U(VI) reduction in
replicated sediment microcosms. Biodegradation 16, 1–14.
Payne R. B., Gentry D. M., Rapp-Giles B. J., Casalot L. and Wall
J. D. (2002) Uranium reduction by Desulfovibrio desulfuricans
strain G20 and a cytochrome c3 mutant. Appl. Environ.
Microbiol. 68, 3129–3132.
Peacock A. D., Chang Y.-J., Istok J. D., Krumholz L.,
Geyer R.,
Kinsall B., Watson D., Sublette K. L. and White D. C.
(2004) Utilization of microbial biofilms as monitors of
bioremediation. Microb. Ecol. 47, 284–292.
Revsbech N. P. (2005) Analysis of microbial communities Reduction behavior of uranium in the presence of
with electroche mical microsensors and microscale citric acid.
biosensors. Meth- ods Enzymol. 397, 147–166. Radiochim. Acta 94, 579–583.
Revsbech N. P. and Jorgensen B. B. (1986) Microelectrodes: their Tebo B. M. and Obraztsova A. Y. (1998) Sulfate-reducing
use in microbial ecology. In Advances in Microbial Ecology, vol. bacterium grows with CR(VI), U(VI), MN(IV), and Fe(III) as
9 (ed. K. C. Marshall). Plenum, New York, pp. 293–352. electron acceptors. FEMS Microbiol. Lett. 162, 193–198.
Rodriguez G. G., Philips D., Ishiguro K. and Ridgway H. F. Tucker M. D., Barton L. L. and Thomason B. M. (1996)
(1992) Use of fluorescent redox probe for direct Kinetic coefficients for simultaneous reduction of sulfate
visualization of actively respiring bacteria. Appl. Environ. and uranium by Desulfovibrio desulfuricans. Appl. Microbiol.
Microbiol. 58(6), Biotechnol. 46,
1801–1808. 74–77.
Sani R. K., Peyton B. M., Smith W. A., Apel W. A. and Petersen J. Tucker M. D., Barton L. L. and Thomason B. M. (1998)
N. (2002) Dissimilatory reduction of Cr(VI), Fe(III), and U(VI) Removal of U and Mo from water by immobilized
by Cellulomonas isolates. Appl. Microbiol. Biotechnol. 60, 192– Desulfovibrio desulfuricans in column reactors. Biotechnol.
199. Bioeng. 60, 88–96. Turner J. S. and Robinson N. J. (1995)
Schleifer K. H., Amann R. I., Ludwig W., Rothemund C., Springer Cyanobacterial metallo- thioneins—biochemistry and molecular-
N. and Dorn S. (1992) Nucleic acid probes for the identification genetics. J. Ind. Micro-
and in situ detection of pseudomonas. In Pseudomonas: biol. 14(2), 119–125.
Molecular Biology and Biotechnology (eds. E. Galli, S. Silver Ullmann W. J. and Aller R. C. (1982) Diffusion
and B. Withold). American Society for Microbiology, Wash- coefficients in nearshore marine sediments. Limnol.
ington, DC, pp. 127–134. Oceanogr. 27, 552–556.
Schmidt A., Haferburg G., Sineriz M., Merten D., Bu¨ chel Vilchez R., Pozo C., Gomez M. A., Rodelas B. and
G. and Kothe E. (2005) Heavy metal resistance Gonzalez- Lopez J. (2007) Dominance of sphingomonads
mechanisms in actino- bacteria for survival in AMD in a copper- exposed biofilm community for groundwater
contaminated soils. Chem. Erde treatment. Micro- biology 153, 325–337.
65, 131– Viret H., Pringault O. and Duran R. (2006) Impact of
144. zinc and nickel on oxygen consumption of benthic
Shelobolina E. S., Sullivan S. A., O’Neill K. R., Nevin K. microbial communi- ties assessed with microsensors. Sci.
P. and Lovley D. R. (2004) Isolation, characterization, Total Environ. 367, 302–
and U(VI)- reducing potential of a facultatively 311.
anaerobic, acid-resistant bacterium from low-pH, Waterhouse S. and Hall G. M. (1995) The validation of
nitrate- and U(VI)-contaminated subsurface sediment and sterilising grade microfiltration membranes with
description of Salmonella subterranea sp. nov. Appl. Environ. Pseudomonas diminuta a review. J. Membr. Sci. 104, 1–9.
Microbiol. 70, 2959–2965. Wimpenny J., Manz W. and Szewzyk U. (2000)
Sprocati A. R., Alisi C., Segre L., Tasso F., Heterogeneity in biofilms. FEMS Microbiol. Rev. 24(5), 661–
Galletti M. and 671.
Cremisini C. (2006) Investigating heavy metal Wu W.-M., Gu B., Fields M. W., Gentile M., Ku Y.-
resistance, bioaccumulation and metabolic profile of a K., Yan H.,
metallophile microbial consortium native to an Tiquias S., Yan T., Nyman J., Zhou J., Jardine P.
abandoned mine. Sci. Total Environ. 366, 649–658. M. and Criddle C. S. (2005) Uranium (VI) reduction
Stohs S. J. and Bagchi D. (1995) Oxidative mechanisms by denitrifying biomass. Bioremed. J. 9, 1–13.
in the toxicity of metal–ions. Free Radic. Biol. Med. 18(2), Xu K. D., Stewart P. S., Xia F., Huang C.-T. and McFeters
321–336. G. A. (1998) Spatial physiological heterogeneity in
Stoodley P., deBeer D. and Lewandowski Z. (1994) Liquid Pseudomonas aeruginosa biofilm is determined by oxygen
flow in availability. Appl. Environ. Microbiol. 64, 4035–4039.
biofilm systems. Appl. Environ. Microbiol. 60, 2711– Yu T. and Bishop P. L. (2001) Stratification and
2716. oxidation– reduction potential change in an aerobic and
Suzuki Y., Kelly S. D., Kemner K. M. and Banfied J. F. (2002) sulfate-reducing biofilm studied using microelectrodes.
Nanometersize products of uranium bioreduction. Nature 419, Water Environ. Res.
134. 73(3), 368–373.
Suzuki Y., Kelly S. D., Kemner K. M. and Banfied J. F. Zinn M. S.,KirkegaardR.D., Palmer, Jr.,R.J. and WhiteD.C.
(2003) Microbial populations stimulated for hexavalent (1999) Laminar flow chamber for continuous monitoring
uranium reduction in uranium mine sediment. Appl. of biofilm formation and succession. Methods Enzymol. 310,
Environ. Microbiol. 224–232.
69, 1337–
1346.
Suzuki Y., Nankawa T., Yoshida T., Ozaki T., Ohnuki T., Associate editor: Roy A. Wogelius
Francis
A. J., Tsushima S., Enokida Y. and Yamamoto I.
(2006)