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Analysis and succession of nitrifying bacteria community structure in sequencing biofilm batch reactor

  • Applied genetics and molecular biotechnology
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Abstract

To reveal the succession procedure of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) community structure in sequencing biofilm batch reactor (SBBR), the molecular biological techniques of denaturing gradient gel electrophoresis (DGGE), cloning, and real-time PCR were applied. DGGE showed that the structural diversity of the bacterial community increased during the biofilm formation period, and some kinds of populations had been highly preponderant consistently. The results of cloning and sequencing revealed that Nitrosomonas was the dominant species. The real-time PCR analysis indicated that the amount of the AOB increased significantly after the cultivation period, and the NOB gradually decreased. The AOB content on the 25th day was 17 times that of the 6th day. It also showed the biofilm formed successfully with accumulating nitrite and prepared to achieve the achievement of simultaneous nitrification and denitrification in SBBR. Furthermore, the ammonia-oxidizing rate was in correspondence with the NH4 +-N removal efficiency.

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References

  • Anthonisen AC, Loehr RC, Prakasam TB, Srinath EG (1976) Inhibition of nitrification by ammonia and nitrous acid. Water Pollut Control Fed 48(5):835–852

    CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • Broude NE (2002) Stem-loop oligonucleotides: a robust tool for molecular biology and biotechnology. Trends Biotechnol 6:249–256

    Article  Google Scholar 

  • Cesar M, Melanie AH, Jennifer AR, Jay JC, de los Reyes FL III (2005) Effects of aeration cycles on nitrifying bacterial populations and nitrogen removal in intermittently aerated reactors. Appl Environ Microbiol 71(12):8565–8572

    Article  CAS  Google Scholar 

  • Dejonghe W, Boon N, Seghers D, Top EM, Verstraete W (2001) Bioaugmentation of soils by increasing microbial richness: missing links. Environ Microbiol 3:649–657

    Article  PubMed  CAS  Google Scholar 

  • Dionisi HM, Layton AC, Harins G, Gregory IR, Robinson KG, Sayler GS (2002) Quantification of Nitrosomonas oligotropha-like ammonia-oxidizing bacteria and Nitrospira spp. from full-scale wastewater treatment plants by competitive PCR. Appl Environ Microbiol 68:245–253

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Fang F, Bing-Jie N, Xiao-Yan L, Guo-Ping S, Han-Qing Y (2009) Kinetic analysis on the two-step processes of AOB and NOB in aerobic nitrifying granules. Appl Microbiol Biotechnol 83:1159–1169

    Article  PubMed  CAS  Google Scholar 

  • George O, Heidi BK, Roberto K (2000) Biofilm formation as microbial development. Annu Rev Microbiol 54:49–79

    Article  Google Scholar 

  • Gerards S, Duyts H, Laanbroek HJ, Gerards S, Duyts H, Laanbroek HJ (1998) Ammonium-induced inhibition of ammonium-starved Nitrosomonas europaea cells in soil and sand slurries. FEMS Microbiol Ecol 26:269–280

    Article  CAS  Google Scholar 

  • Gerda H, Alice CL, Hebe MD, Igrid RG, Victoria MG, Shawn AH, Kevin GR, Gary SS (2003) Real-time PCR quantification of nitrifying bacteria in a municipal wastewater treatment plant. Environ Sci Technol 37:343–351

    Article  CAS  Google Scholar 

  • Hanaki K, Chalermraj W, Shinichiro O (1990) Nitrification at low levels of dissolved oxygen with and without organic loading in a suspended-growth reactor. Water Res 24:297–302

    Article  CAS  Google Scholar 

  • Jae-Hoon C, Sang L, Kensuke F, Kazuo Y (2007) Comparison of sludge characteristics and PCR–DGGE based microbial diversity of nanofiltration and microfiltration membrane bioreactors. Chemosphere 67:1543–1550

    Article  CAS  Google Scholar 

  • Juan CL, Sonia H, Stephanie H, Fabrice ML, Gerard C, Laurent P (2004) Quantification of a novel group of nitrate-reducing bacteria in the environment by real-time PCR. J Microbiol Methods 57(3):399–407

    Article  CAS  Google Scholar 

  • Jubany I, Baeza JA, Carrera J, Lafuente J (2005) Respirometric calibration and validation of a biological nitrite oxidation model including biomass growth and substrate inhibition. Water Res 39:4574–4584

    Article  PubMed  CAS  Google Scholar 

  • Kindaichi T, Ito T, Okabe S (2004) Ecophysiological interaction between nitrifying bacteria and heterotrophic bacteria in autotrophic nitrifying biofilms as determined by microautoradiography-fluorescence in situ hybridization. Appl Environ Microbiol 70(3):1641–1650

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Kowalchuk GA, Stephen JR, De BOER W, Prosser JI, Embley TM, Woldendorp JW (1997) Analysis of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in coastal sand dunes by denaturing gradient gel electrophoresis and sequencing of PCR-amplified 16S ribosomal DNA fragments. Appl Environ Microbiol 63(4):1489–1497

    PubMed Central  PubMed  CAS  Google Scholar 

  • Limpiyakom T, Shinohara Y, Kurisu F, Yag O (2005) Communities of ammonia-oxidizing bacteria in activated sludge of various sewage treatment plants in Tokyo. FEMS Microbiol Ecol 54(2):205–217

    Article  CAS  Google Scholar 

  • Limpiyakorn T, Kurisu F, Yagi O (2006) Development and application of real-time PCR for quantification of specific ammonia-oxidizing bacteria in activated sludge of sewage treatment systems. Appl Microbiol Biotechnol 72:1004–1013

    Article  PubMed  CAS  Google Scholar 

  • Ling W, Tian L (2011) Anaerobic ammonium oxidation in constructed wetlands with biocontact oxidation as pretreatment. Ecol Eng 37:1225–1230

    Article  Google Scholar 

  • Livak KJ, Flood SJA, Marmaro J, Giusti W, Deetz K (1995) Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Methods Appl 4:357–362

    Article  PubMed  CAS  Google Scholar 

  • Marion DE, Guy S, Fabrice ML (2004) Real-time reverse transcription PCR analysis of expression of atrazine catabolism genes in two bacterial strains isolated from soil. J Microbiol Methods 56(1):3–15

    Article  CAS  Google Scholar 

  • Nicolaisen MH, Ramsing NB (2002) Denaturing gradient gel electrophoresis (DGGE) approaches to study the diversity of ammonia-oxidizing bacteria. J Microbiol Methods 50(2):189–203

    Article  PubMed  CAS  Google Scholar 

  • Nubel U, Engelen B, Felske A, Snaidr J, Wieshuber A, Amann RI, Ludwig W, Backhaus H (1996) Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. J Bacteriol 178:5636–5643

    PubMed Central  PubMed  CAS  Google Scholar 

  • Thomas E, Freitag, Lisa C, Christopher DC, James I (2005) Influence of inorganic nitrogen management regime on the diversity of nitrite-oxidizing bacteria in agricultural grassland soils. Appl Environ Microbiol 71(12):8323–8334

    Article  CAS  Google Scholar 

  • Tyagi S, Kramer FR (1996) Molecular beacons: probes that fluoresce upon hybridization. Nat Biotechnol 14:303–308

    Article  PubMed  CAS  Google Scholar 

  • Wiesmann U (1994) Biological nitrogen removal from wastewater. Adv Biochem Eng Biotechnol 51:113–154

    PubMed  CAS  Google Scholar 

  • Ying Y, Zhenmei LV, Mim H, Cheng J (2008) Dynamic changes of microbial community diversity in a photohydrogen producing reactor monitored by PCR-DGGE. J Environ Sci 20:1118–1125

    Article  CAS  Google Scholar 

  • Yutaka O, Krassimira RH, Christian ML, Louise EJ, De Ford R, Binyam G, David L, Kate MS (2003) Application of real-time PCR to study effects of ammonium on population size of ammonia-oxidizing bacteria in soil. Appl Environ Microbiol 70(2):1008–1016

    Google Scholar 

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (grant no. 51178125) and S&T Planned Project of Guangdong Province (grant no. 2012B030800005).

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Authors and Affiliations

  1. Key Laboratory for Water Quality Security and Protection in Pearl River Delta, Ministry of Education and Guangdong Province, Guangzhou University, Guangzhou, 510006, China

    Jingyin Wang, Chaosheng Zhang & Hongwei Rong

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  1. Jingyin Wang
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  2. Chaosheng Zhang
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  3. Hongwei Rong
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Correspondence to Chaosheng Zhang.

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Wang, J., Zhang, C. & Rong, H. Analysis and succession of nitrifying bacteria community structure in sequencing biofilm batch reactor. Appl Microbiol Biotechnol 98, 4581–4587 (2014). https://doi.org/10.1007/s00253-014-5537-6

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  • DOI: https://doi.org/10.1007/s00253-014-5537-6

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