Optimization and Application of Fluorescence in Situ Hybridization Assay for Detecting Polyphosphate - Accumulating Microorganisms

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Laboratory-scale sequencing batch reactors (SBRs) were operated on activated sludge processes were used to study enhanced biological phosphorus removal (EBPR) from wastewater. Polyphosphate-accumulating microorganisms (PAOs) play an important role during the enhanced biological phosphorus removal (EBPR) process. Fluorescence in situ hybridization (FISH) was applied to assess the proportions of microorganisms in the sludge. The aim of this study was to optimize hybridization of PAOMIX and RHC439 probes by orthogonal design. Orthogonal optimization test of the four factors were conducted under the individual three levels. The optimal hybridizition conditions were as follow: hybridization temperature 46°C, hybridization time 2.5h, washing time 15min, formamide concentration 35%(PAOMIX probe); hybridization temperature 50°C, hybridization time 2.5h, washing time 20min, formamide concentration 20% (RHC439 probe).

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Advanced Materials Research (Volumes 183-185)

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1369-1373

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January 2011

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© 2011 Trans Tech Publications Ltd. All Rights Reserved

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[1] Y.H. Kong, J.L. Nielsen and P.H. Nielsen: Appl. Environ. Microbiol. Vol. 70(2004), pp.5383-5390.

Google Scholar

[2] M. Pijuan, A.M. Saunders, A. Guisasola, J.A. Baeza, C. Casas and L.L. Blackall: Biotechnol. Bioeng. Vol. 85(2004), pp.56-67.

DOI: 10.1002/bit.10813

Google Scholar

[3] K.D. McMahon M.A. Dojka N.R. Pace,D. Jenkins and J.D. Keasling: Appl. Environ. Microbiol. Vol. 68(2004), pp.4971-4978.

Google Scholar

[4] C.M. Lopez-Vazquez, Y.I. Song, C.M. Hooijmans, D. Brdjanovic, M.S. Moussa, H.J. Gijzen and M.C. M van Loosdrecht: Biotechnol. Bioeng. Vol. 101(2008), pp.295-306.

DOI: 10.1002/bit.21892

Google Scholar

[5] Y. Zhou, M. Pijuan, R.J. Zeng, H.B. Lu and Z.G. Yuan: Water Res. Vol. 42(2008), pp.2361-2368.

Google Scholar

[6] R.J. Zeng, A.M. Saunders, Z.G. Yuan L.L. Blackall and J. Keller: Biotechnol. Bioeng. Vol. 83(2003), pp.140-148.

Google Scholar

[7] A. Oehmen, R.J. Zeng, Z.G. Yuan, and J. Keller: Biotechnol. Bioeng. Vol. 91(2005), pp.43-53.

Google Scholar

[8] H.B. Lu, A. Oehmen,B. Virdis, J. Keller and Z.G. Yuan: Water Res. Vol. 40(2006), pp.3838-3848.

Google Scholar

[9] H.S. Park, R. Schumacher and J. J. Kilbane:J. Ind. Microbiol. Biotechnol. Vol. 32 (2005), pp.94-102.

Google Scholar

[10] A. Moter and U.B. Göbel: J. Microbiol. Methods, Vol. 41 (2000), p.85–112.

Google Scholar

[11] K. Soejima, K. Oki , A. Terada , S. Tsuneda and A. Hirata: Bioprocess and Biosystems Engineering  Vol. 29 (2006), pp.305-313.

DOI: 10.1007/s00449-006-0079-7

Google Scholar

[12] G.J.F. Smolders, J. Vandermeij, M.C.M. van Loosdrecht and J.J. Heijnen: Biotechnol. Bioeng. Vol. 43 (1994), pp.461-470.

Google Scholar

[13] P.L. Bond, , R. Erhart, M. Wagner, J. Keller, and L.L. Blackall: Appl. Environ. Microbiol. Vol. 65 (1999), pp.4077-4084.

Google Scholar

[14] R.I. Amann, In: Akkermans ADC, van Elsas JD, de Brujin FJ (eds) Molecular microbial ecology manual. Kluwer, Dordrecht (1995).

Google Scholar

[15] H. Daims, A. Bruhl, R. Amann, K. -H. Schleifer and M. Wagner: Syst. Appl. Microbiol. Vol. 22 (1999), pp.434-444.

Google Scholar

[16] G.R. Crocetti, P. Hugenholtz, P.L. Bond, A. Schuler, J. Keller, D Jenkins and L.L. Blackall: Appl. Environ. Microbiol.  Vol. 66 (2000), pp.1175-1182.

DOI: 10.1128/aem.66.3.1175-1182.2000

Google Scholar

[17] R.I. Amann, L. Krumholz and D.A. Stahl: J. Bacteriol. Vol. 172 (1990), p.762–770.

Google Scholar

[18] R.P.X. Hesselmann, C. Werlen, D. Hahn, J. R. Van Der Meer and A.J.B. Zehnder: Syst. Appl. Microbiol. Vol. 22 (1999), pp.454-465.

Google Scholar

[19] Y.H. Wang, Zh.T. Feng and J.P. Merlio, in: Theory and Technology of Molecular Hybridization, edtied by Science Press, Bei Jing (2005), in press.

Google Scholar

[20] M. Meganck, D. Malnou and P. LeFlohic: Water Sci. Technol. Vol. 17 (1985), pp.199-121.

Google Scholar