Microbial community variation and functions to excess sludge reduction in a novel gravel contact oxidation reactor.

Excess biomass produced within the degradation processes of organic pollutants is creating environmental challenges. The gravel contact oxidation reactor (GCOR) filled with crushed stone globular aggregates as carriers, has been demonstrated capable of reducing the excess sludge effectively in some pilot and small-scale engineering studies. In order to evaluate the variation and structure of the microbial community and their functions to excess sludge reduction in GCOR, a conventional activated sludge reactor (ASR) was studied as a comparison. The 16S rDNA library of the universal bacteria was constructed, Shannon's diversity index (H) and Species evenness (E) were calculated with distance-based operational taxonomic unit and richness (DOTUR) for microbial diversity. Real-time quantity PCR and optical microscope were used for absolute bacterial DNA concentration and eukarya identification, respectively. Meanwhile, the suspended solid index in GCOR and ASR was detected for assessing the excess sludge production. The results indicated that the most abundant bacteria in GCOR were those related to the beta-Proteobacteria group, then gamma-Proteobacteria and to Cytophaga-Flexibacter-Bacteriode (CFB). In the ASR samples major bacteria were in the closest match with gamma-Proteobacteria, then beta-Proteobacteria and CFB. Shannon's index (H) was higher (3.41) for diversity of bacteria extracted from the carrier samples in GCOR than that (2.71) from the sludge sample in ASR. Species evenness (E) for the isolates from GCOR and ASR samples was 0.97 and 0.96, respectively. Comparison of the universal bacteria population in GCOR and ASR shows that the total bacterial DNA concentration on the GCOR carriers were 8.98 x 10(5) microg/ microl, twice that in ASR of 4.67 x 10(5) microg/ microl under normal operation of two reactors. But the MLSS in GCOR was only 4.5mg/L, 25 times less than that in ASR of 115.4 mg/L. The most representative eukarya were protozoa both in GCOR (15 no. per 20 ml) and in ASR (15 no. per 20 ml); the next abundant group was attachment plants 10 no. per 20 ml in GCOR and 4 no. per 20 ml in ASR, respectively. Rotifers and copepoda belonging to metazoan were only present in GCOR (8 no. per 20 ml for both rotifers and copepoda). The microbial diversity and population difference both in the GCOR carriers and ASR sludge indicated that the diverse microbes, a large amount of biomass forming longer microbial food chains attached on the carriers may be the main functions for the excess sludge reduction in GCOR.

[1]  P. Hugenholtz,et al.  The use of 16S rDNA clone libraries to describe the microbial diversity of activated sludge communities , 1998 .

[2]  G. Hamer,et al.  Death and lysis during aerobic thermophilic sludge treatment: Characterization of recalcitrant products , 1994 .

[3]  P. Madoni,et al.  Toxic effect of heavy metals on the activated sludge protozoan community , 1996 .

[4]  E. Topp,et al.  Development of a rapid quantitative PCR assay for direct detection and quantification of culturable and non-culturable Escherichia coli from agriculture watersheds. , 2007, Journal of microbiological methods.

[5]  Dick B Janssen,et al.  Efficient recovery of environmental DNA for expression cloning by indirect extraction methods. , 2003, FEMS microbiology ecology.

[6]  Aditi A. Moharikar,et al.  A novel approach for extraction of PCR-compatible DNA from activated sludge samples collected from different biological effluent treatment plants. , 2003, Journal of microbiological methods.

[7]  J. Handelsman,et al.  Introducing DOTUR, a Computer Program for Defining Operational Taxonomic Units and Estimating Species Richness , 2005, Applied and Environmental Microbiology.

[8]  R. E. Buchanan,et al.  Bergey's Manual of Determinative Bacteriology. , 1975 .

[9]  M. Wagner,et al.  The microbial community composition of a nitrifying-denitrifying activated sludge from an industrial sewage treatment plant analyzed by the full-cycle rRNA approach. , 2002, Systematic and applied microbiology.

[10]  J. Harel,et al.  Optimization of microbial DNA extraction and purification from raw wastewater samples for downstream pathogen detection by microarrays. , 2005, Journal of microbiological methods.

[11]  C. Jeanthon,et al.  Microbial diversity in production waters of a low-temperature biodegraded oil reservoir. , 2005, FEMS microbiology ecology.

[12]  C. Amblard,et al.  Changes in the structure and metabolic activities of periphytic communities in a stream receiving treated sewage from a waste stabilization pond , 1998 .

[13]  H Geissler,et al.  Estimation of the growth rate of mixed ruminal bacteria from short-term DNA radiolabeling. , 1998, Anaerobe.

[14]  Guang-Hao Chen,et al.  Possible cause of excess sludge reduction in an oxic-settling-anaerobic activated sludge process (OSA process). , 2003, Water research.

[15]  L. Paulin,et al.  Paucibacter toxinivorans gen. nov., sp. nov., a bacterium that degrades cyclic cyanobacterial hepatotoxins microcystins and nodularin. , 2005, International journal of systematic and evolutionary microbiology.

[16]  T. García,et al.  Real-time PCR for detection and quantification of red deer (Cervus elaphus), fallow deer (Dama dama), and roe deer (Capreolus capreolus) in meat mixtures. , 2008, Meat science.

[17]  E. Stackebrandt,et al.  The phylogenetic position of Serratia, Buttiauxella and some other genera of the family Enterobacteriaceae. , 1999, International journal of systematic bacteriology.

[18]  R Amann,et al.  Phylogenetic analysis and in situ identification of bacteria in activated sludge , 1997, Applied and environmental microbiology.

[19]  A. Sadaie,et al.  Reducing Sludge Production and the Domination of Comamonadaceae by Reducing the Oxygen Supply in the Wastewater Treatment Procedure of a Food-Processing Factory , 2007, Bioscience, biotechnology and biochemistry.

[20]  Willy Verstraete,et al.  Reduced sludge production in a two-stage membrane-assisted bioreactor , 2000 .

[21]  Ji‐Zheng He,et al.  Pre-lysis washing improves DNA extraction from a forest soil , 2005 .

[22]  A. Magurran Ecological Diversity and Its Measurement , 1988, Springer Netherlands.

[23]  M. Kimura,et al.  DGGE method for analyzing 16S rDNA of methanogenic archaeal community in paddy field soil. , 2004, FEMS microbiology letters.

[24]  W. D. de Vos,et al.  Response of a Soil Bacterial Community to Grassland Succession as Monitored by 16S rRNA Levels of the Predominant Ribotypes , 2000, Applied and Environmental Microbiology.

[25]  Thomas P. Curtis,et al.  Uncoupling of metabolism to reduce biomass production in the activated sludge process , 2000 .

[26]  H. Chase,et al.  Reducing production of excess biomass during wastewater treatment , 1999 .