Journal of Applied & Environmental Microbiology
ISSN (Print): 2373-6747 ISSN (Online): 2373-6712 Website: http://www.sciepub.com/journal/jaem Editor-in-chief: Sankar Narayan Sinha
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Journal of Applied & Environmental Microbiology. 2014, 2(4), 176-184
DOI: 10.12691/jaem-2-4-11
Open AccessArticle

An Application of Sequencing Batch Reactors in the Identification of Microbial Community Structure from an Activated Sludge

M. Shah1,

1Industrial Waste Water Research Laboratory Division of Applied & Environmental Microbiology Enviro Technology Limited Gujarat, India

Pub. Date: June 15, 2014

Cite this paper:
M. Shah. An Application of Sequencing Batch Reactors in the Identification of Microbial Community Structure from an Activated Sludge. Journal of Applied & Environmental Microbiology. 2014; 2(4):176-184. doi: 10.12691/jaem-2-4-11

Abstract

In this study, the bacterial community structures of phosphate- and non-phosphate-removing activated sludges were compared. Sludge samples were obtained from two sequencing batch reactors (SBRs), and 16S rDNA clone libraries of the bacterial sludge populations were established. Community structures were determined by phylogenetic analyses of 97 and 92 partial clone sequences from SBR1 (phosphate-removing sludge) and SBR2 (non-phosphate-removing sludge), respectively. For both sludges, the predominant bacterial group with whichclones were affiliated was the beta subclass of the proteobacteria. Other major groups represented were the alpha proteobacterial subclass, planctomycete group, and Flexibacter-Cytophaga-Bacteroides group. In addition, several clone groups unaffiliated with known bacterial assemblages were identified in the clone libraries. Acinetobacter spp., thought to be important in phosphate removal in activated sludge, were poorly represented by clone sequences in both libraries. Differences in community structure were observed between the phosphate and non-phosphate-removing sludges; in particular, the Rhodocyclus group within the beta subclass was represented to a greater extent in the phosphate-removing community. Such differences may account for the differing phosphate-removing capabilities of the two activated sludge communities.

Keywords:
activated sludge rhodocyclus acinetobacter phosphate

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References:

[1]  Seviour R, Nielsen PH (2010) Microbial ecology of activated sludge. London: IWA Publishing Company. 688 p.
 
[2]  Bitton G (2011) Wastewater microbiology. Hoboken, NJ: John Wiley and Sons. 746 p.
 
[3]  Graham DW, Smith VH (2004) Designed ecosystem services: Application of ecological principles in wastewater treatment engineering. Front Ecol Environ 4: 199-206.
 
[4]  Wang X, Wen X, Yan H, Ding K, Zhao F, et al. (2011) Bacterial community dynamics in a functionally stable pilot-scale wastewater treatment plant. Bioresour Technol 102: 2352-2357.
 
[5]  Curtis TP, Head IM, Graham DW (2003) Theoretical ecology in engineering biology. Environ Sci Technol 37: 64A-70A.
 
[6]  Wells GF, Park HD, Eggleston B, Francis CA, Criddle CS (2011) Fine-scale bacterial community dynamics and the taxa-time relationship within a full-scale activated sludge bioreactor. Water Res 45: 5476-5488.
 
[7]  Wagner M, Loy A, Nogueira R, Purkhold U, Lee N, et al. (2002) Microbial community composition and function in wastewater treatment plants. Antonie van Leeuwenhoek 81: 665-680.
 
[8]  Kaewpipat K, Grady CPL (2002) Microbial population dynamics in laboratory scale activated sludge reactors. Water Sci Technol 46: 19-27.
 
[9]  Padayachee P, Ismail A, Bux F (2006) Elucidation of the microbial community structure within a laboratory-scale activated sludge process using molecular techniques. Water SA 32: 679-686.
 
[10]  Jenkins D (2008) From total suspended solids to molecular biology toolsa personal view of biological wastewater treatment process population dynamics. Water Environ Res 80: 677-687.
 
[11]  Saikaly PE, Stroot PG, Oerther DB (2005) Use of 16S rRNA gene terminal restriction fragment analysis to assess the impact of solids retention time on the bacterial diversity of activated sludge. Appl Environ Microbiol 71: 5814-5822.
 
[12]  Ofit¸eru ID, Lunn M, Curtis TP, Wells GF, Criddle CS, et al. (2010) Combined niche and neutral effects in a microbial wastewater treatment community. Proc Natl Acad Sci U S A 107: 15345-15350.
 
[13]  Sanapareddy N, Hamp TJ, Gonzalez LC, Hilger HA, Fodor AA, et al. (2009) Molecular diversity of a North Carolina wastewater treatment plant as revealed by pyrosequencing. Appl Environ Microbiol 75: 1688-1696.
 
[14]  Fierer N, Lennon JT (2011) The generation and maintenance of diversity in microbial communities. Am J Bot 98: 439-448.
 
[15]  Nemergut DR, Costello EK, Hamady M, Lozupone C, Jiang L, et al. (2011) Global patterns in the biogeography of bacterial taxa. Environ Microbiol 13: 135-144.
 
[16]  Benedict RG, Carlson DA (1971) Aerobic heterotrophic bacteria in activated sludge. Water Res 5: 1023-1030.
 
[17]  Dias FF, Bhat JV (1964) Microbial ecology of activated sludge. Appl Microbiol 12: 412-417.
 
[18]  Lighthart B, Oglesby RT (1969) Bacteriology of an activated sludge wastewater treatment plant: A guide to methodology. J Water Pollut Control Fed 41: R267-R281.
 
[19]  van Veen W (1973) Bacteriology of activated sludge, in particular the filamentous bacteria. Antonie van Leeuwenhoek 39: 189-205.
 
[20]  Eschenhagen M, Schuppler M, Ro¨ske I (2003) Molecular characterization of the microbial community structure in two activated sludge systems for the advanced treatment of domestic effluents. Water Res 37: 3224-3232.
 
[21]  Snaidr J, Amann R, Huber I, Ludwing W, Schleifer K (1997) Phylogenetic analysis and in-situ identification of bacteria in activated sludge. Appl Environ Microbiol 63: 2884-2896.
 
[22]  Wagner M, Amann R, Lemmer H, Schleifer K (1993) Probing activated sludge with oligonucleotides specific for proteobacteria: Inadequacy of culturedependent methods for describing microbial community structure. Appl Environ Microbiol 59: 1520-1525.
 
[23]  Watanabe K, Yamamoto S, Hino S, Harayama S (1998) Population dynamics of phenol-degrading bacteria in activated sludge determined by gyrB-targeted quantitative PCR. Appl Environ Microbiol 64: 1203-1209.
 
[24]  Green JL, Bohannan BJM (2006) Spatial scaling of microbial biodiversity. Trends Ecol Evol 21: 501-507.
 
[25]  Martiny JBH, Bohannan BJM, Brown JH, Colwell RK, Fuhrman JA, et al. (2006) Microbial biogeography: putting microorganisms on the map. Nat Rev Microbiol 4: 102-112.
 
[26]  van der Gast CJ, Jefferson B, Reid E, Robinson T, Bailey MJ, et al. (2006) Bacterial diversity is determined by volume in membrane bioreactors. Environ Microbiol 8: 1048-1055.
 
[27]  Wang X, Wen X, Criddle C, Wells G, Zhang J, et al. (2010) Community analysis of ammonia-oxidizing bacteria in activated sludge of eight wastewater treatment systems. J Environ Sci (China) 22: 627-634.
 
[28]  Loy A, Daims H, Wagner M (2002) Activated sludge: molecular techniques for determining community composition. In: Bitton G, editor. The Encyclopedia of environmental microbiology. Hoboken, NJ: John Wiley and Sons. 26-43.
 
[29]  Onuki M, Satoh H, Mino T, Matsuo T (2000) Application of molecular methods to microbial community analysis of activated sludge. Water Sci Technol 42: 17-22.
 
[30]  Wilderer PA, Bungartz HJ, Lemmer H, Wagner M, Keller J, et al. (2002) Modern scientific methods and their potential in wastewater science and technology. Water Res 36: 370-393.
 
[31]  de los Reyes FL III (2010) Challenges in determining causation in structurefunction studies using molecular biological techniques. Water Res 44: 4948-4957.
 
[32]  Jones PA, Schuler AJ (2010) Seasonal variability of biomass density and activated sludge settleability in full-scale wastewater treatment systems. Chem Eng J 164: 16-22.
 
[33]  Carvalho G, Lemos PC, Oehmen A, Reis MAM (2007) Denitrifying phosphorus removal: Linking the process performance with the microbial community structure. Water Res 41: 4383-4396.
 
[34]  Briones A, Raskin L (2003) Diversity and dynamics of microbial communities in engineered environments and their implications for process stability. Curr Opin Biotechnol 14: 270-276.
 
[35]  Gentile ME, Jessup CM, Nyman JL, Criddle CS (2007) Correlation of functional instability and community dynamics in denitrifying dispersed-growth reactors. Appl Environ Microbiol 73: 680-690.
 
[36]  Curtis TP, Sloan WT (2006) Towards the design of diversity: stochastic models for community assembly in wastewater treatment plants. Water Sci Technol 54: 227-236.
 
[37]  Daims H, Taylor MW, Wagner M (2006) Wastewater treatment: a model system for microbial ecology. Trends Biotechol 24: 483-489.
 
[38]  Prosser JI, Bohannan BJ, Curtis TP, Ellis RJ, Firestone MK, et al. (2007) The role of ecological theory in microbial ecology. Nat Rev Microbiol 5: 384-392.
 
[39]  Lane, D. J. (1991). 16S/23S rRNA sequencing, p. 115-175. In E. Stackebrandt and M. Goodfellow (ed.), Nucleic acid techniques in bacterial systematics. John Wiley & Sons, New York.
 
[40]  Sambrook, J., E. F. Fritsch, and T. Maniatis. (1989). Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
 
[41]  Lane, D. J. (1991). 16S/23S rRNA sequencing, p. 115-175. In E. Stackebrandt and M. Goodfellow (ed.), Nucleic acid techniques in bacterial systematics. John Wiley & Sons, New York.
 
[42]  32. Larsen, N., G. J. Olsen, B. L. Maidak, M. J. McCaughey, R. Overbeek, T. J. Macke, T. L. Marsh, and C. R. Woese. (1993). The ribosomal database project. Nucleic Acids Res. 21: 3021-3023.
 
[43]  Blackall, L. L., S. C. Barker, and P. Hugenholtz. (1994). Phylogenetic analysis and taxonomic history of Nocardia pinensis and Nocardia amarae. Syst. Appl. Microbiol. 17: 519-525.
 
[44]  Mino, T., T. Kawakami, and T. Matsuo. (1985). Behaviour of intracellular polyphosphate in the biological phosphate removal process. Water Sci. Technol. 17 (12): 11-21.
 
[45]  Toerien, D. F., A. Gerber, L. H. Lo¨tter, and T. E. Cloete. (1990). Enhanced biological phosphorus removal in activated sludge systems. Adv. Microb. Ecol. 11: 173-230.
 
[46]  Liesack, W., R. So¨ller, T. Stewart, H. Haas, S. Giovannoni, and E. Stackebrandt. (1992). The influence of tachytelically (rapidly) evolving sequences on the topology of phylogenetic trees intrafamily relationships and the phylogenetic position of Planctomycetaceae as revealed by comparative analysis of 16S ribosomal RNA sequences. Syst. Appl. Microbiol. 15: 357-362.
 
[47]  Bayly, R. C., J. W. May, W. G. C. Raper, A. Duncan, N. H. Pilkington, and G. E. Vasiliadis. (1989). The effect of primary fermentation on biological nutrient removal, p. 162-166. In Australian Water and Wastewater Association 13th Federal Convention. Australian Water and Wastewater Association, Canberra, Australia.
 
[48]  Lo¨tter, L. H., and A. P. Pitman. (1992). Improved biological phosphorus removal resulting from the enrichment of reactor feed with fermentation products. Water Sci. Technol. 26 (5-6): 943-953.
 
[49]  Winter, C. T. (1989). The role of acetate in denitrification and biological phosphate removal in modified bardenpho systems. Water Sci. Technol. 21 (4-9): 375-385.
 
[50]  Brock, T. D. (1987). The study of microorganisms in situ: progress and problems. Symp. Soc. Gen. Microbiol. 41: 1-17.
 
[51]  Manz, W., M. Wagner, R. Amann, and K. H. Schleifer. (1994). In situ characterization of the microbial consortia active in two wastewater treatment plants. Water Res. 28: 1715-1723.
 
[52]  Wagner, M., R. Amann, H. Lemmer, and K. H. Schleifer. (1993). Probing activated sludge with oligonucleotides specific for proteobacteria: inadequacy of culture-dependent methods for describing microbial community structure. Appl. Environ. Microbiol. 59: 1520-1525.
 
[53]  DeLong, E. F., D. G. Franks, and A. L. Alldredge. (1993). Phylogenetic diversity of aggregate-attached vs. free-living marine bacterial assemblages. Liminol. Oceanogr. 38: 924-934.
 
[54]  16. Ekendahl, S., J. Arlinger, F. Stahl, and K. Pedersen. (1994). Characterisation of attached bacterial populations in deep granitic groundwater from the Stripa research mine by 16S rRNA gene sequencing and scanning electron microscopy. Microbiology 140: 1575-1583.
 
[55]  Liesack, W., H. Weyland, and E. Stackebrandt. (1991). Potential risks of gene amplification by PCR as determined by 16S rDNA analysis of a mixedculture of strict barophilic bacteria. Microb. Ecol. 21: 191-198.
 
[56]  Pace, N. R., D. A. Stahl, D. J. Lane, and G. J. Olsen. (1986). The analysis of natural microbial populations by ribosomal RNA sequences. Adv. Microb. Ecol. 9: 1-55.
 
[57]  Schmidt, T. M., E. F. DeLong, and N. R. Pace. (1991). Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J. Bacteriol. 173: 4371-4378.
 
[58]  Straub, T. M., I. L. Pepper, M. Abbaszadegan, and C. P. Gerba. (1994). A method to detect enteroviruses in sewage sludge-amended soil using the PCR. Appl. Environ. Microbiol. 60: 1014-1017.
 
[59]  Emberly, T. M., R. P. Hirt, and D. M. Williams. (1994). Biodiversity at the molecular level: the domains, kingdoms and phyla of life. Philos. Trans. R. Soc. London B Biol. Sci. 345: 21-33.
 
[60]  Wentzel, M. C., R. E. Loewenthal, G. A. Ekama, and G. R. Marais. (1988). Enhanced polyphosphate organism cultures in activated sludge systems. Part 1. Enhanced culture development. Water S. A. (Pretoria) 14: 81-92.
 
[61]  Hiraishi, A., K. Masamune, and H. Kitamura. (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: 897-901.
 
[62]  25. Hiraishi, A., and Y. Morishima. (1990). Capacity for polyphosphate accumulation of predominant bacteria in activated sludge showing enhanced phosphate removal. J. Ferment. Bioeng. 69: 368-371.
 
[63]  Wagner, M., R. Erhart, W. Manz, R. Amann, H. Lemmer, D. Wedi, and K. H. Schleifer. (1994). Development of an rRNA-targeted oligonucleotide probe specific for the genus Acinetobacter and its application for in situ monitoring in activated sludge. Appl. Environ. Microbiol. 60: 792-800.
 
[64]  Larsen, N., G. J. Olsen, B. L. Maidak, M. J. McCaughey, R. Overbeek, T. J. Macke, T. L. Marsh, and C. R. Woese. (1993). The ribosomal database project. Nucleic Acids Res. 21: 3021-3023.
 
[65]  Hiraishi, A., and H. Kitamura. (1984). Distribution of phototrophic purple nonsulfur bacteria in activated sludge systems and other aquatic environments. Bull. Jpn. Soc. Sci. Fish. 50: 1929-1937.
 
[66]  Shin, Y. K., A. Hiraishi, and J. Sugiyama. (1993). Molecular systematics of the genus Zoogloea and emendation of the genus. Int. J. Syst. Bacteriol. 43: 826-831.
 
[67]  van Niekerk, A. M., D. Jenkins, and M. D. Richard. (1987). The competitive growth of Zoogloea ramigera and Type 021N in activated sludge and pure culture
 
[68]  Schuppler, M., F. Mertens, G. Scho¨n, and U. B. Go¨bel. (1995). Molecular characterization of nocardiform actinomycetes in activated sludge by 16S rRNA analysis. Microbiology 141: 513-521.
 
[69]  Siefert, E., R. L. Irgens, and N. Pfennig. (1978). Phototrophic purple and green bacteria in a sewage treatment plant. Appl. Environ. Microbiol. 35: 38-44.
 
[70]  Hiraishi, A., J. L. Shi, and H. Kitamura. (1989). Effects of organic nutrient strength on the purple nonsulfur bacterial content and metabolic activity of photosynthetic sludge for wastewater treatment. J. Ferment. Bioeng. 68: 269-276.
 
[71]  Toerien, D. F., A. Gerber, L. H. Lo¨tter, and T. E. Cloete. (1990). Enhanced biological phosphorus removal in activated sludge systems. Adv. Microb. Ecol. 11: 173-230.
 
[72]  Hiraishi, A., and H. Kitamura. (1984). Differences in phototrophic growth on high phosphate concentrations among Rhodopseudomonas species. J. Ferment. Technol. 62: 293-296.
 
[73]  Hiraishi, A., A. Yanase, and H. Kitamura. (1991). Polyphosphate accumulation by Rhodobacter sphaeroides grown under different environmental conditions with special emphasis on the effect of external phosphate concentrations. Bull. Jpn. Soc. Microb. Ecol. 6: 25-32.
 
[74]  Kristiansen, J. (1971). On Planctomyces bekefii and its occurrence in Danish lakes and ponds. Bot. Tidsskr. 66: 293-302.
 
[75]  Schlesner, H. (1994). The development of media suitable for the microorganisms morphologically resembling Planctomyces spp., Pirellula spp., and other Planctomycetales from various aquatic habitats using dilute media. Syst. Appl. Microbiol. 17: 135-145.
 
[76]  Hiraishi, A. (1988). Respiratory quinone profiles as tools for identifying different bacterial populations in activated sludge. J. Gen. Appl. Microbiol. 34: 39-56.
 
[77]  Takeuchi, J. (1991). Influence of nitrate on the bacterial flora of activated sludge under anoxic condition. Water Sci. Technol. 23 (4-6): 765-772.
 
[78]  Cech, J. S., P. Hartman, and J. Wanner. (1993). Competition between PolyP and non-PolyP bacteria in an enhanced phosphate removal system. Water Environ. Res. 65: 690-693.
 
[79]  Brodisch, K. E. U. (1985). Interaction of different groups of microorganisms in biological phosphate removal. Water Sci. Technol. 17 (12): 89-97.
 
[80]  8. Buchan, L. (1983). Possible biological mechanism of phosphorus removal. Water Sci. Technol. 15 (3-4): 87-103.
 
[81]  Bux, F., and H. C. Kasan. (1994). A microbiological survey of ten activated sludge plants. Water S. A. (Pretoria) 20: 61-72.
 
[82]  Pickup, R. W. (1991). Development of molecular methods for the detection of specific bacteria in the environment. J. Gen. Microbiol. 137: 1009-1019.
 
[83]  Trebesius, K., R. Amann, W. Ludwig, K. Mu¨hlegger, and K. H. Schleifer. (1994). Identification of whole fixed bacterial cells with nonradioactive 23S rRNA-targeted polynucleotide probes. Appl. Environ. Microbiol. 60: 3228-3235.