Tetrasphaera have been recently identified based on the 16S ribosomal RNA(rRNA)gene as among the most abundant polyphosphate-accumulating organisms(PAOs)in global full-scale wastewater treatment plants(WWTPs)with enha...Tetrasphaera have been recently identified based on the 16S ribosomal RNA(rRNA)gene as among the most abundant polyphosphate-accumulating organisms(PAOs)in global full-scale wastewater treatment plants(WWTPs)with enhanced biological phosphorus removal(EBPR).However,it is unclear how Tetrasphaera PAOs are selectively enriched in the context of the EBPR microbiome.In this study,an EBPR microbiome enriched with Tetrasphaera(accounting for 40%of 16S sequences on day 113)was built using a top-down design approach featuring multicarbon sources and a low dosage of allylthiourea.The microbiome showed enhanced nutrient removal(phosphorus removal~85%and nitrogen removal~80%)and increased phosphorus recovery(up to 23.2 times)compared with the seeding activated sludge from a local full-scale WWTP.The supply of 1 mg·L^(-1)allylthiourea promoted the coselection of Tetrasphaera PAOs and Microlunatus PAOs and sharply reduced the relative abundance of both ammonia oxidizer Nitrosomonas and putative competitors Brevundimonas and Paracoccus,facilitating the establishment of the EBPR microbiome.Based on 16S rRNA gene analysis,a putative novel PAO species,EBPR-ASV0001,was identified with Tetrasphaera japonica as its closest relative.This study provides new knowledge on the establishment of a Tetrasphaera-enriched microbiome facilitated by allylthiourea,which can be further exploited to guide future process upgrading and optimization to achieve and/or enhance simultaneous biological phosphorus and nitrogen removal from high-strength wastewater.展开更多
To elucidate the phosphorus removal and metabolism under various COD / P ratio,a sludge highly enriched in PAOs was used to investigate the impacts of COD / P in batch tests under different carbon supply conditions. A...To elucidate the phosphorus removal and metabolism under various COD / P ratio,a sludge highly enriched in PAOs was used to investigate the impacts of COD / P in batch tests under different carbon supply conditions. Acetate,propionate and a mixture of acetate and propionate at a ratio of 3 ∶ 1( COD basis) was used as carbon sources with the COD / P of 20,15,10 and 5. 0 g COD /gP,respectively. The minimum COD / P ratios for complete P removal were found to be 8. 24 g COD /gP for acetate,11. 40 g COD /gP for propionate and9. 10 g COD /gP for the 3 ∶ 1 mixture of acetate and propionate. Converted to a mass basis,all three cases had a very similar ratio of 7. 7 g VFA /gP,which represented a useful guide for operation of EBPR plants to identify possible shortages in VFAs. The trend in PHV accumulation during the anaerobic period along with the decrease of COD / P ratios suggested that,PAOs may use the TCA pathway for anaerobic VFA uptake to maintain the required NADH production with reduced glycogen degradation. During the aerobic phase,the glycogen pool was reduced but remained enough compared to the requirement for anaerobic VFA uptake,and the synthesis and degradation of glycogen was not the inhibition factor of PAOs.展开更多
Enhanced biological phosphorus removal(EBPR) was investigated in an anaerobic/aerobic sequencing batch reactor(SBR) supplied with glucose as a single organic substrate.The results illustrated that EBPR process could a...Enhanced biological phosphorus removal(EBPR) was investigated in an anaerobic/aerobic sequencing batch reactor(SBR) supplied with glucose as a single organic substrate.The results illustrated that EBPR process could also occur successfully with glucose other than short chain fatty acids(SCFAs).High phosphorus release and poly-hydroxyalkanoate(PHA) accumulation in the anaerobic phase was found vital for the removal of phosphorus during the aerobic phase.The measurement of intracellular reserves revealed that glycogen had a higher chance to replace the energy role of poly-P under anaerobic conditions.Moreover,glycogen was also utilized as the carbon source for PHA synthesis,as well as a reducing power as reported earlier.The accumulated PHA in this system was mainly in the form of poly-hydroxyvalerate(PHV) instead of poly-hydroxybutyrate(PHB),and was inferred to be caused by the excess reducing power contained in glucose.Lactate as a fermentation product was also found released into the bulk solution.Applying fundamental biochemistry knowledge to the experimental results,a conceptual biochemical model was developed to explain the metabolism of the glucose-induced EBPR.展开更多
Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were stu...Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were studied.Most YSR-3 cells stored one or two polyP granules in regular appearance and high-density.The diameter of the granules was about 400 nm measuring by a transmission electron microscope(TEM).After stained with 4,6-diamidino-2-phenylindole(DAPI)and visualized by a fluorescence microscope,the cells turned blue and the granules were bright yellow.The composition of granules includes P(major ingredient),Mg,S,K,and Ca as detected by an energy dispersive X-ray spectrometer(EDS).When inorganic phosphorus(po34-)and ferric ion(Fe3+)were added into media,the biomass increased and the cells formed intracellular polyP granules owing to the phosphorus assimilation from media.The YSR-3 obtained higher biomass by adding 0.02 g/L FePO4 than 0.005 g/L and 0.01 g/L FePO4;however,the phosphorus absorption was higher with 0.01 g/L FePO4 than 0.005 g/L and 0.02 g/L FePO4.The optical density at wavelength 480 nm(OD480nm)was 0.79 and 100%cells could form intracellular polyP granules.These results show that strain YSR-3 is able to acquire higher biomass and absorb more inorganic phosphorus when 0.01 g/L FePO4 is added.The characteristics of absorbing and storing phosphorus as intracellular inorganic polyP granules have a potential for application in high-efficiency phosphorus removal in wastewater treatment.展开更多
In this study, the influence of azo dye of methyl red (MR) on COD, dye and phosphorus removal and the transformation of polyhydroxyalkanoate (PHA) and glycogen of phosphate accumulating organisms in enhanced biologica...In this study, the influence of azo dye of methyl red (MR) on COD, dye and phosphorus removal and the transformation of polyhydroxyalkanoate (PHA) and glycogen of phosphate accumulating organisms in enhanced biological phosphorus removal (EBPR) system were investigated. The results indicated COD and dye removal efficiencies were decreased from 97.9% to 72.8% and 99.7% to 82.0%, respectively, when MR concentration was increased from 0 to 40 mg/L. Low MR concentration (5 mg/L) had no influence on P removal and transformation of PHA and glycogen. However, P removal, PHA production and consumption, and glycogen replenishment were seriously inhibited at high MR concentration, while glycogen hydrolysis was simulated at MR concentration of 20 and 40 mg/L. The transformations of PHA and glycogen at aerobic condition were more sensitive to those at anaerobic condition at high MR concentration. These results demonstrated dye and its intermediate products would inhibit the metabolism of polyphosphate accumulating organisms, which should be taken into account in future work.展开更多
Nitrous oxide （N2O） emission has been reported to be enhanced during denitrification when internally-stored compounds are used as carbon sources. However, negligible N2O emissions have been detected in the few studi...Nitrous oxide （N2O） emission has been reported to be enhanced during denitrification when internally-stored compounds are used as carbon sources. However, negligible N2O emissions have been detected in the few studies where polyhydroxyalkanoates （PHA） were specifically used. This study investigated and compared the potential enhancement of N2O production, based on utilization of an internally-stored polymer and external carbon （acetate） by a denitrifying phosphorus removal culture. Results indicated that at relatively low chemical oxygen demand-to-nitrogen （COD/N） ratios, more nitrite was reduced to N2O in the presence of an external carbon source as compared to an internal carbon source （PHA）. At relatively higher COD/N ratios, similar N2O reduction rates were obtained in all cases regardless of the type of carbon source available. N2O reduction rates were, however, generally higher in the presence of an internal carbon source. Results from the study imply that when the presence of an external carbon source is not sufficient to support denitrification, it is likely competitively utilized by different metabolic pathways of denitrifying polyphosphate accumulating organisms （DPAOs） and other ordinary denitfifiers. This study also reveals that the consumption of PHA is potentially the rate-limiting step for N2O reduction during denitrification.展开更多
基金supported by the Key Research and Development Program of Zhejiang(2022C03075)National Natural Science Foundation of China(22241603)Zhejiang Provincial Natural Science Foundation of China(LR22D010001)。
文摘Tetrasphaera have been recently identified based on the 16S ribosomal RNA(rRNA)gene as among the most abundant polyphosphate-accumulating organisms(PAOs)in global full-scale wastewater treatment plants(WWTPs)with enhanced biological phosphorus removal(EBPR).However,it is unclear how Tetrasphaera PAOs are selectively enriched in the context of the EBPR microbiome.In this study,an EBPR microbiome enriched with Tetrasphaera(accounting for 40%of 16S sequences on day 113)was built using a top-down design approach featuring multicarbon sources and a low dosage of allylthiourea.The microbiome showed enhanced nutrient removal(phosphorus removal~85%and nitrogen removal~80%)and increased phosphorus recovery(up to 23.2 times)compared with the seeding activated sludge from a local full-scale WWTP.The supply of 1 mg·L^(-1)allylthiourea promoted the coselection of Tetrasphaera PAOs and Microlunatus PAOs and sharply reduced the relative abundance of both ammonia oxidizer Nitrosomonas and putative competitors Brevundimonas and Paracoccus,facilitating the establishment of the EBPR microbiome.Based on 16S rRNA gene analysis,a putative novel PAO species,EBPR-ASV0001,was identified with Tetrasphaera japonica as its closest relative.This study provides new knowledge on the establishment of a Tetrasphaera-enriched microbiome facilitated by allylthiourea,which can be further exploited to guide future process upgrading and optimization to achieve and/or enhance simultaneous biological phosphorus and nitrogen removal from high-strength wastewater.
基金Sponsored by the National Natural Science Foundation of China(Grant No.51278143,51408162)the Postdoctoral Foundation of China(Grant No.2013M530161)
文摘To elucidate the phosphorus removal and metabolism under various COD / P ratio,a sludge highly enriched in PAOs was used to investigate the impacts of COD / P in batch tests under different carbon supply conditions. Acetate,propionate and a mixture of acetate and propionate at a ratio of 3 ∶ 1( COD basis) was used as carbon sources with the COD / P of 20,15,10 and 5. 0 g COD /gP,respectively. The minimum COD / P ratios for complete P removal were found to be 8. 24 g COD /gP for acetate,11. 40 g COD /gP for propionate and9. 10 g COD /gP for the 3 ∶ 1 mixture of acetate and propionate. Converted to a mass basis,all three cases had a very similar ratio of 7. 7 g VFA /gP,which represented a useful guide for operation of EBPR plants to identify possible shortages in VFAs. The trend in PHV accumulation during the anaerobic period along with the decrease of COD / P ratios suggested that,PAOs may use the TCA pathway for anaerobic VFA uptake to maintain the required NADH production with reduced glycogen degradation. During the aerobic phase,the glycogen pool was reduced but remained enough compared to the requirement for anaerobic VFA uptake,and the synthesis and degradation of glycogen was not the inhibition factor of PAOs.
基金Science and Technology Project of Zhejiang and Hangzhou (No2007C13081, No20062912A06)
文摘Enhanced biological phosphorus removal(EBPR) was investigated in an anaerobic/aerobic sequencing batch reactor(SBR) supplied with glucose as a single organic substrate.The results illustrated that EBPR process could also occur successfully with glucose other than short chain fatty acids(SCFAs).High phosphorus release and poly-hydroxyalkanoate(PHA) accumulation in the anaerobic phase was found vital for the removal of phosphorus during the aerobic phase.The measurement of intracellular reserves revealed that glycogen had a higher chance to replace the energy role of poly-P under anaerobic conditions.Moreover,glycogen was also utilized as the carbon source for PHA synthesis,as well as a reducing power as reported earlier.The accumulated PHA in this system was mainly in the form of poly-hydroxyvalerate(PHV) instead of poly-hydroxybutyrate(PHB),and was inferred to be caused by the excess reducing power contained in glucose.Lactate as a fermentation product was also found released into the bulk solution.Applying fundamental biochemistry knowledge to the experimental results,a conceptual biochemical model was developed to explain the metabolism of the glucose-induced EBPR.
基金Supported by the National Natural Science Foundation of China(Nos.51308245,31570061)the Open Project of Jiangsu Provincial Engineering Laboratory for Advanced Materials of Salt Chemical Industry(No.SF201408)the Open Project of Jiangsu Provincial Key Construction Laboratory of Probiotics Preparation(No.JSYSZJ2017006)
文摘Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were studied.Most YSR-3 cells stored one or two polyP granules in regular appearance and high-density.The diameter of the granules was about 400 nm measuring by a transmission electron microscope(TEM).After stained with 4,6-diamidino-2-phenylindole(DAPI)and visualized by a fluorescence microscope,the cells turned blue and the granules were bright yellow.The composition of granules includes P(major ingredient),Mg,S,K,and Ca as detected by an energy dispersive X-ray spectrometer(EDS).When inorganic phosphorus(po34-)and ferric ion(Fe3+)were added into media,the biomass increased and the cells formed intracellular polyP granules owing to the phosphorus assimilation from media.The YSR-3 obtained higher biomass by adding 0.02 g/L FePO4 than 0.005 g/L and 0.01 g/L FePO4;however,the phosphorus absorption was higher with 0.01 g/L FePO4 than 0.005 g/L and 0.02 g/L FePO4.The optical density at wavelength 480 nm(OD480nm)was 0.79 and 100%cells could form intracellular polyP granules.These results show that strain YSR-3 is able to acquire higher biomass and absorb more inorganic phosphorus when 0.01 g/L FePO4 is added.The characteristics of absorbing and storing phosphorus as intracellular inorganic polyP granules have a potential for application in high-efficiency phosphorus removal in wastewater treatment.
文摘In this study, the influence of azo dye of methyl red (MR) on COD, dye and phosphorus removal and the transformation of polyhydroxyalkanoate (PHA) and glycogen of phosphate accumulating organisms in enhanced biological phosphorus removal (EBPR) system were investigated. The results indicated COD and dye removal efficiencies were decreased from 97.9% to 72.8% and 99.7% to 82.0%, respectively, when MR concentration was increased from 0 to 40 mg/L. Low MR concentration (5 mg/L) had no influence on P removal and transformation of PHA and glycogen. However, P removal, PHA production and consumption, and glycogen replenishment were seriously inhibited at high MR concentration, while glycogen hydrolysis was simulated at MR concentration of 20 and 40 mg/L. The transformations of PHA and glycogen at aerobic condition were more sensitive to those at anaerobic condition at high MR concentration. These results demonstrated dye and its intermediate products would inhibit the metabolism of polyphosphate accumulating organisms, which should be taken into account in future work.
文摘Nitrous oxide （N2O） emission has been reported to be enhanced during denitrification when internally-stored compounds are used as carbon sources. However, negligible N2O emissions have been detected in the few studies where polyhydroxyalkanoates （PHA） were specifically used. This study investigated and compared the potential enhancement of N2O production, based on utilization of an internally-stored polymer and external carbon （acetate） by a denitrifying phosphorus removal culture. Results indicated that at relatively low chemical oxygen demand-to-nitrogen （COD/N） ratios, more nitrite was reduced to N2O in the presence of an external carbon source as compared to an internal carbon source （PHA）. At relatively higher COD/N ratios, similar N2O reduction rates were obtained in all cases regardless of the type of carbon source available. N2O reduction rates were, however, generally higher in the presence of an internal carbon source. Results from the study imply that when the presence of an external carbon source is not sufficient to support denitrification, it is likely competitively utilized by different metabolic pathways of denitrifying polyphosphate accumulating organisms （DPAOs） and other ordinary denitfifiers. This study also reveals that the consumption of PHA is potentially the rate-limiting step for N2O reduction during denitrification.
