Candidatus Accumulibacter phosphatis: Elusive Bacterium Responsible for Enhanced Biological Phosphorus Removal
Publication: Journal of Environmental Engineering
Volume 140, Issue 1
Abstract
Enhanced biological phosphorus removal (EBPR) is an effective nutrient removal process that has been engineered and successfully applied in wastewater treatment for decades. Although the prevalent microorganism performing EBPR in full-scale activated sludge plants was identified nearly a decade ago as Candidatus Accumulibacter phosphatis, it still remains an unculturable organism. In this paper, the authors describe research that has been aimed at identifying and isolating microorganisms responsible for EBPR, along with the fundamental metabolic, genetic, and ecological advances that have been recently made in the characterization of Cand. A. phosphatis. Furthermore, evidence is presented that Cand. A. phosphatis can perform the EBPR cycle under nitrogen-limiting conditions, a characteristic that could be potentially incorporated into strategies to obtain pure cultures of this organism. Finally, possible research directions that could be enabled when a pure culture of this organism is obtained were discussed.
Get full access to this article
View all available purchase options and get full access to this article.
Acknowledgments
This research was partially funded by a Paul L. Busch award from the Water Environment Research Foundation and by Associated Engineering (Burnaby, British Columbia, Canada). We thank Katherine D. McMahon for providing samples from Cand. A. phosphatis enrichment cultures.
References
Albertsen, M., Hansen, L. B. S., Saunders, A. M., Nielsen, P. H., and Nielsen, K. L. (2012). “A metagenome of a full-scale microbial community carrying out enhanced biological phosphorus removal.” ISME J., 6(6), 1094–1106.
Blackall, L. L., Seviour, E. M., Bradford, D., Rossetti, S., Tandoi, V., and Seviour, R. J. (2000). “Candidatus Nostocoida limicola, a filamentous bacterium from activated sludge.” Int. J. Syst. Evol. Microbiol., 50(2), 703–709.
Bond, P. L., Hugenholtz, P., Keller, J., and Blackall, L. L. (1995). “Bacterial community structures of phosphate-removing and non-phosphate-removing activated sludges from sequencing batch reactors.” Appl. Environ. Microbiol., 61(5), 1910–1916.
Brodisch, K. E. U. (1985). “Interaction of different groups of microorganisms in biological phosphate removal.” Water Sci. Technol., 17(11–1), 89–97.
Brown, M. V., Schwalbach, M. S., Hewson, I., and Fuhrman, J. A. (2005). “Coupling 16S-Its rDNA clone libraries and automated ribosomal intergenic spacer analysis to show marine microbial diversity: Development and application to a time series.” Environ. Microbiol., 7(9), 1466–1479.
Buchan, L. (1983). “Possible biological mechanism of phosphorus removal.” Water Sci. Technol., 15(3–4), 87–103.
Burow, L. C., Mabbett, A. N., and Blackall, L. L. (2008). “Anaerobic glyoxylate cycle activity during simultaneous utilization of glycogen and acetate in uncultured Accumulibacter enriched in enhanced biological phosphorus removal communities.” ISME J., 2(10), 1040–1051.
Carvalho, G., Lemos, P. C., Oehmen, A., and Reis, M. A. M. (2007). “Denitrifying phosphorus removal: Linking the process performance with the microbial community structure.” Water Res., 41(19), 4383–4396.
Cloete, T. E., and Steyn, P. L. (1988). “A combined membrane filter-immunofluorescent technique for the in situ identification and enumeration of Acinetobacter in activated sludge.” Water Res., 22(8), 961–969.
Crocetti, G. R., et al. (2000). “Identification of polyphosphate-accumulating organisms and design of 16S rRNA-directed probes for their detection and quantitation.” Appl. Environ. Microbiol., 66(3), 1175–1182.
Crocetti, G. R., Banfield, J. F., Keller, J., Bond, P. L., and Blackall, L. L. (2002). “Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processes.” Microbiology, 148(11), 3353–3364.
Deinema, M. H., Van Loosdrecht, M., and Scholten, A. (1985). “Some physiological-characteristics of Acinetobacter spp accumulating large amounts of phosphate.” Water Sci. Technol., 17(11–1), 119–125.
Dixon, R., and Kahn, D. (2004). “Genetic regulation of biological nitrogen fixation.” Nat. Rev. Microbiol., 2(8), 621–631.
Erhart, R., Bradford, D., Seviour, R. J., Amann, R., and Blackall, L. L. (1997). “Development and use of fluorescent in situ hybridization probes for the detection and identification of Microthrix parvicella in activated sludge.” Syst. Appl. Microbiol., 20(2), 310–318.
Fisher, M. M., and Triplett, E. W. (1999). “Automated approach for ribosomal intergenic spacer analysis of microbial diversity and its application to freshwater bacterial communities.” Appl. Environ. Microbiol., 65(10), 4630–4636.
Flowers, J. J., et al. (2013). “Comparative genomics of two Candidatus Accumulibacter clades performing biological phosphorus removal.” ISME J.,.
Flowers, J. J., He, S. M., Yilmaz, S., Noguera, D. R., and McMahon, K. D. (2009). “Denitrification capabilities of two biological phosphorus removal sludges dominated by different Candidatus Accumulibacter clades.” Environ. Microbiol. Rep., 1(6), 583–588.
Fuhs, G. W., and Chen, M. (1975). “Microbiological basis of phosphate removal in the activated sludge process for treatment of wastewater.” Microb. Ecol., 2(2), 119–138.
Goel, R. K., Sanhueza, P., and Noguera, D. R. (2005). “Evidence of Dechloromonas sp. participating in enhanced biological phosphorus removal (EBPR) in a bench-scale aerated anoxic reactor.” Water Environment Federation 78th Annual Technical Exhibition and Conf., Water Environmenta Federation, Washington, DC, 3864–3871.
Hanada, S., Liu, W. T., Shintani, T., Kamagata, Y., and Nakamura, K. (2002). “Tetrasphaera elongata sp nov., a polyphosphate-accumulating bacterium isolated from activated sludge.” Int. J. Syst. Evol. Microbiol., 52(3), 883–887.
He, S., Gall, D. L., and McMahon, K. D. (2007). “Candidatus Accumulibacter population structure in enhanced biological phosphorus removal sludges as revealed by polyphosphate kinase genes.” Appl. Environ. Microbiol., 73(18), 5865–5874.
He, S., Gu, A. Z., and McMahon, K. D. (2006). “Fine-scale differences between Accumulibacter-like bacteria in enhanced biological phosphorus removal activated sludge.” Water Sci. Technol., 54(1), 111–117.
He, S. M., et al. (2010a). “Metatranscriptomic array analysis of Candidatus Accumulibacter phosphatis-enriched enhanced biological phosphorus removal sludge.” Environ. Microbiol., 12(5), 1205–1217.
He, S. M., Bishop, F. I., and McMahon, K. D. (2010b). “Bacterial community and Candidatus Accumulibacter population dynamics in laboratory-scale enhanced biological phosphorus removal reactors.” Appl. Environ. Microbiol., 76(16), 5479–5487.
He, S. M., and McMahon, K. D. (2011). “Candidatus Accumulibacter gene expression in response to dynamic EBPR conditions.” ISME J., 5(2), 329–340.
Hesselmann, R. P. X., Von Rummell, R., Resnick, S. M., Hany, R., and Zehnder, A. J. B. (2000). “Anaerobic metabolism of bacteria performing enhanced biological phosphate removal.” Water Res., 34(14), 3487–3494.
Hesselmann, R. P. X., Werlen, C., Hahn, D., van der Meer, J. R., and Zehnder, A. J. B. (1999). “Enrichment, phylogenetic analysis and detection of a bacterium that performs enhanced biological phosphate removal in activated sludge.” Syst. Appl. Microbiol., 22(3), 454–465.
Hien, T. T. N., Le, V. Q., Hansen, A. A., Nielsen, J. L., and Nielsen, P. H. (2011). “High diversity and abundance of putative polyphosphate-accumulating tetrasphaera-related bacteria in activated sludge systems.” FEMS Microbiol. Ecol., 76(2), 256–267.
Hiraishi, A., Masamune, K., and Kitamura, H. (1989). “Characterization of the bacterial population structure in an anaerobic–aerobic activated sludge system on the basis of respiratory quinone profiles.” Appl. Environ. Microbiol., 55(4), 897–901.
Jenkins, D., and Tandoi, V. (1991). “The applied microbiology of enhanced biological phosphate removal—accomplishment and needs.” Water Res., 25(12), 1471–1478.
Kang, D.-W., and Noguera, D. R. (2011). “Development of a novel strategy to cultivate enhanced biological phosphorus removal (EBPR) microorganisms.” Water Environment Federation 84th Annual Technical Exhibition and Conf., Water Environment Federation, Los Angeles.
Kiley, P. J., and Donohue, T. J. (2011). “Global responses of bacteria to oxygen deprivation.” Bacterial stress responses, G. Storz and R. Hengge, eds., ASM Press, Washington, DC, 175–204.
Kim, J. M., Lee, H. J., Lee, D. S., and Jeon, C. O. (2013). “Characterization of the denitrification-associated phosphorus uptake properties of Candidatus Accumulibacter phosphatis clades in sludge subjected to enhanced biological phosphorus removal.” Appl. Environ. Microbiol., 79(6), 1969–1979.
Kong, Y. H., Nielsen, J. L., and Nielsen, P. H. (2004). “Microautoradiographic study of Rhodocyclus-related polyphosphate accumulating bacteria in full-scale enhanced biological phosphorus removal plants.” Appl. Environ. Microbiol., 70(9), 5383–5390.
Kong, Y. H., Nielsen, J. L., and Nielsen, P. H. (2005). “Identity and ecophysiology of uncultured actinobacterial polyphosphate-accumulating organisms in full-scale enhanced biological phosphorus removal plants.” Appl. Environ. Microbiol., 71(7), 4076–4085.
Kong, Y. H., Xia, Y., Nielsen, J. L., and Nielsen, P. H. (2007). “Structure and function of the microbial community in a full-scale enhanced biological phosphorus removal plant.” Microbiology, 153(12), 4061–4073.
Kristiansen, R., et al. (2013). “A metabolic model for members of the genus Tetrasphaera involved in enhanced biological phosphorus removal.” ISME J., 7(3), 543–554.
Liu, W. T., Nielsen, A. T., Wu, J. H., Tsai, C. S., Matsuo, Y., and Molin, S. (2001). “In situ identification of polyphosphate- and polyhydroxyalkanoate-accumulating traits for microbial populations in a biological phosphorus removal process.” Environ. Microbiol., 3(2), 110–122.
Lu, H. B., Oehmen, A., Virdis, B., Keller, J., and Yuan, Z. G. (2006). “Obtaining highly enriched cultures of Candidatus Accumulibacter phosphates through alternating carbon sources.” Water Res., 40(20), 3838–3848.
Majed, N., Chernenko, T., Diem, M., and Gu, A. Z. (2012). “Identification of functionally relevant populations in enhanced biological phosphorus removal processes based on intracellular polymers profiles and insights into the metabolic diversity and heterogeneity.” Environ. Sci. Technol., 46(9), 5010–5017.
Marques, R., et al. (2013). “Characterisation of the ecological niches in enhanced biological phosphorus removal processes with Tetrasphaera and Accumulibacter.” 5th Int. Conf.—Microbial Ecology and Water Engineering, International Water Association, Ann Arbor, MI.
Martin, H. G., et al. (2006). “Metagenomic analysis of two enhanced biological phosphorus removal (EBPR) sludge communities.” Nat. Biotechnol., 24(10), 1263–1269.
Maszenan, A. M., et al. (2000). “Three isolates of novel polyphosphate-accumulating Gram-positive cocci, obtained from activated sludge, belong to a new genus, Tetrasphaera gen. nov., and description of two new species, Tetrasphaera japonica sp, nov and Tetrasphaera australiensis sp nov.” Int. J. Syst. Evol. Microbiol., 50(2), 593–603.
Maurer, M., Gujer, W., Hany, R., and Bachmann, S. (1997). “Intracellular carbon flow in phosphorus accumulating organisms from activated sludge systems.” Water Res., 31(4), 907–917.
McIlroy, S. J., et al. (2013). “Metabolic model for the filamentous Candidatus Microthrix parvicella based on genomic and metagenomic analyses.” ISME J., 7(6), 1161–1172.
McMahon, K. D., Dojka, M. A., Pace, N. R., Jenkins, D., and Keasling, J. D. (2002). “Polyphosphate kinase from activated sludge performing enhanced biological phosphorus removal.” Appl. Environ. Microbiol., 68(10), 4971–4978.
Mielczarek, A. T., Nguyen, H. T. T., Nielsen, J. L., and Nielsen, P. H. (2013). “Population dynamics of bacteria involved in enhanced biological phosphorus removal in Danish wastewater treatment plants.” Water Res., 47(4), 1529–1544.
Mino, T., Van Loosdrecht, M. C. M., and Heijnen, J. J. (1998). “Microbiology and biochemistry of the enhanced biological phosphate removal process.” Water Res., 32(11), 3193–3207.
Nakamura, K., Hiraishi, A., Yoshimi, Y., Kawaharasaki, M., Masuda, K., and Kamagata, Y. (1995). “Microlunatus phosphovorus gen-nov, sp-nov, a new Gram-positive polyphosphate-accumulating bacterium isolated from activated-sludge.” Int. J. Syst. Bacteriol., 45(1), 17–22.
Nguyen, H. T. T., Le, V. Q., Hansen, A. A., Nielsen, J. L., and Nielsen, P. H. (2011). “High diversity and abundance of putative polyphosphate-accumulating Tetrasphaera-related bacteria in activated sludge systems.” FEMS Microbiol. Ecol., 76(2), 256–267.
Oehmen, A., et al. (2007). “Advances in enhanced biological phosphorus removal: from micro to macro scale.” Water Res., 41(11), 2271–2300.
Oehmen, A., Carvalho, G., Freitas, F., and Reis, M. A. M. (2010). “Assessing the abundance and activity of denitrifying polyphosphate accumulating organisms through molecular and chemical techniques.” Water Sci. Technol., 61(8), 2061–2068.
Perez-Feito, R., Peccia, J., and Noguera, D. R. (2006). “Comparison between direct microscopy and flow cytometry for rRNA-based quantification of Candidatus Accumulibacter phosphatis in activated sludge.” Water Environ. Res., 78(2), 181–188.
Peterson, S. B., Warnecke, F., Madejska, J., McMahon, K. D., and Hugenholtz, P. (2008). “Environmental distribution and population biology of Candidatus Accumulibacter, a primary agent of biological phosphorus removal.” Environ. Microbiol., 10(10), 2692–2703.
Satoh, H., Mino, T., and Matsuo, T. (1998). “Anaerobic uptake of glutamate and aspartate by enhanced biological phosphorus removal activated sludge.” Water Sci. Technol., 37(4–5), 579–582.
Seviour, R. J., Mino, T., and Onuki, M. (2003). “The microbiology of biological phosphorus removal in activated sludge systems.” FEMS Microbiol. Rev., 27(1), 99–127.
Stante, L., Cellamare, C. M., Malaspina, F., Bortone, G., and Tilche, A. (1997). “Biological phosphorus removal by pure culture of Lampropedia spp.” Water Res., 31(6), 1317–1324.
Streichan, M., Golecki, J. R., and Schon, G. (1990). “Polyphosphate-accumulating bacteria from sewage plants with different processes for biological phosphorus removal.” FEMS Microbiol. Ecol., 73(2), 113–124.
Tandoi, V., Majone, M., May, J., and Ramadori, R. (1998). “The behaviour of polyphosphate accumulating acinetobacter isolates in an anaerobic–aerobic chemostat.” Water Res., 32(10), 2903–2912.
Unden, G., and Bongaerts, J. (1997). “Alternative respiratory pathways of Escherichia coli: energetics and transcriptional regulation in response to electron acceptors.” Biochimica Et Biophysica Acta-Bioenergetics, 1320(3), 217–234.
Wagner, M., et al. (1994). “Development of an ribosomal-rna-targeted oligonucleotide probe specific for the genus Acinetobacter and its application for in-situ monitoring in activated-sludge.” Appl. Environ. Microbiol., 60(3), 792–800.
Wexler, M., Richardson, D. J., and Bond, P. L. (2009). “Radiolabelled proteomics to determine differential functioning of Accumulibacter during the anaerobic and aerobic phases of a bioreactor operating for enhanced biological phosphorus removal.” Environ. Microbiol., 11(12), 3029–3044.
Wilmes, P., et al. (2008). “Community proteogenomics highlights microbial strain-variant protein expression within activated sludge performing enhanced biological phosphorus removal.” ISME J., 2(8), 853–864.
Wright, E. S., Yilmaz, L. S., and Noguera, D. R. (2012). “Decipher, a search-based approach to chimera identification for 16S rRNA sequences.” Appl. Environ. Microbiol., 78(3), 717–725.
Zilles, J. L., Hung, C. H., and Noguera, D. R. (2002a). “Presence of Rhodocyclus in a full-scale wastewater treatment plant and their participation in enhanced biological phosphorus removal.” Water Sci. Technol., 46(1–2), 123–128.
Zilles, J. L., Peccia, J., Kim, M. W., Hung, C. H., and Noguera, D. R. (2002b). “Involvement of Rhodocyclus-related organisms in phosphorus removal in full-scale wastewater treatment plants.” Appl. Environ. Microbiol., 68(6), 2763–2769.
Zilles, J. L., Peccia, J., and Noguera, D. R. (2002c). “Microbiology of enhanced biological phosphorus removal in aerated-anoxic orbal processes.” Water Environ. Res., 74(5), 428–436.
Information & Authors
Information
Published In
Journal of Environmental Engineering
Volume 140 • Issue 1 • January 2014
Pages: 2 - 10
Copyright
© 2014 American Society of Civil Engineers.
History
Received: Apr 15, 2013
Accepted: Sep 4, 2013
Published online: Sep 6, 2013
Published in print: Jan 1, 2014
Discussion open until: Feb 6, 2014
Authors
Metrics & Citations
Metrics
Citations
Download citation
If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download.