extracellular polymeric substances (EPSs) excreted by bacteria andform firmly attached initial granules [11]. The selective pressure,chemical oxygen demand (COD) loading rate, and settling time ofthe divalent metal ions and many other factors affect the start-upof aerobic granulation [12–15].
The presence of divalent metal ions, such as Ca2+and Mg2+, is animportant factor that reinforces the granulation through the strongeffect of the ions on the self-immobilization of microbial biomassduring start-up aerobic granulation. EPSs would rather bind withmultivalent metals to form stable complexes; e.g., Ca2+affects thephysical characteristics of aerobic granules, while the addition ofMg2+increases microbial diversity in mature granules [15,16].Previous studies showed that iron (Fe2+) increases the granula-tion of anaerobic granular sludge in UASB reactors [17]. The effectsof zero-valent iron (ZVI) on anaerobic granular sludge were alsoinvestigated. The observations are explained by (i) the ZVI reac-tion benefiting the growth of methanogens through acid bufferingand the reduction of the oxidation–reduction potential and (ii)Fe2+leaching from ZVI advancing the granule aggregation [18].However, there is no information on the effects of ZVI on aerobicgranulation.
Iron, an essential element for nearly all organisms, participatesin most important metabolic reactions, such as the syntheses ofhemoglobin, iron-sulfur protein, and other iron-bearing proteins.The iron metal is of low cost, is easy to obtain, and emerging effec-tively degraded contaminants [19]. Additionally, the use of ZVI haslow operation and maintenance costs compared with the use ofother divalent metal ions. For example, nitrate can be convertedto ammonium ion, and ammonia can then be stripped under astrong alkaline condition; hence the total aqueous nitrogen amountcan be reduced by adding ZVI. Moreover, some major reactions,which involve nitrate reduction, ammonia production, and ammo-nia stripping, can be modeled by pseudo first-order kinetics. ZVIhas been investigated as a new medium for the treatment of con-taminated water [20]. Thus, the use of ZVI as a reactive medium forwastewater treatment is a promising technique.We assumed the enhancement of aerobic granulation via ZVIreaction. This study investigates the effects of ZVI on aerobic gran-ulation in a sequencing batch airlift reactor (SBAR). The start-uptime and physical and chemical properties of aerobic granulationwere determined. Furthermore, EPSs were studied using three-dimensional excitation emission matrix (3D-EEM) fluorescencespectra technology targeting the fluorescence of EPSs and com-ponents. Polymerase chain reaction (PCR) denaturing gradient gelelectrophoresis (DGGE) technology was employed to reveal thestructure of the microbial community. Hence, this work could beuseful for the development of aerobic granulation systems and thewide application of ZVI in the field of biotechnology. To the best ofour knowledge, this is the first attempt to enhance aerobic granu-lation by adding ZVI.
2. Materials and methods
2.1. Experimental set-up and operationExperiments were performed in parallel using two double-walled, cylindrical column SBARs with internal diameter of 8 cm.The SBARs contained an internal riser (bottom clearance of 5 cm,internal diameter of 1.5 cm, and height of 80 cm) with a workingvolume of 3.6 L. Air was introduced via a fine bubble aerator througha porous stone diffuser at the bottom of the reactor at an airflowrate of 3 L per minute during the aeration phase.Both reactors were operated in successive cycles with each cyclelasting 6 h. One cycle consisted of five successive phases:(1)10 minfeeding,(2)310 min aerobic reaction,(3)10 min settling,(4)5 mindischarging
extracellular polymeric substances (EPSs) excreted by bacteria andform firmly attached initial granules [11]. The selective pressure,chemical oxygen demand (COD) loading rate, and settling time ofthe divalent metal ions and many other factors affect the start-upof aerobic granulation [12–15].The presence of divalent metal ions, such as Ca2+and Mg2+, is animportant factor that reinforces the granulation through the strongeffect of the ions on the self-immobilization of microbial biomassduring start-up aerobic granulation. EPSs would rather bind withmultivalent metals to form stable complexes; e.g., Ca2+affects thephysical characteristics of aerobic granules, while the addition ofMg2+increases microbial diversity in mature granules [15,16].Previous studies showed that iron (Fe2+) increases the granula-tion of anaerobic granular sludge in UASB reactors [17]. The effectsof zero-valent iron (ZVI) on anaerobic granular sludge were alsoinvestigated. The observations are explained by (i) the ZVI reac-tion benefiting the growth of methanogens through acid bufferingand the reduction of the oxidation–reduction potential and (ii)Fe2+leaching from ZVI advancing the granule aggregation [18].However, there is no information on the effects of ZVI on aerobicgranulation.Iron, an essential element for nearly all organisms, participatesin most important metabolic reactions, such as the syntheses ofhemoglobin, iron-sulfur protein, and other iron-bearing proteins.The iron metal is of low cost, is easy to obtain, and emerging effec-tively degraded contaminants [19]. Additionally, the use of ZVI haslow operation and maintenance costs compared with the use ofother divalent metal ions. For example, nitrate can be convertedto ammonium ion, and ammonia can then be stripped under astrong alkaline condition; hence the total aqueous nitrogen amountcan be reduced by adding ZVI. Moreover, some major reactions,which involve nitrate reduction, ammonia production, and ammo-nia stripping, can be modeled by pseudo first-order kinetics. ZVIhas been investigated as a new medium for the treatment of con-taminated water [20]. Thus, the use of ZVI as a reactive medium forwastewater treatment is a promising technique.We assumed the enhancement of aerobic granulation via ZVIreaction. This study investigates the effects of ZVI on aerobic gran-ulation in a sequencing batch airlift reactor (SBAR). The start-uptime and physical and chemical properties of aerobic granulationwere determined. Furthermore, EPSs were studied using three-dimensional excitation emission matrix (3D-EEM) fluorescencespectra technology targeting the fluorescence of EPSs and com-ponents. Polymerase chain reaction (PCR) denaturing gradient gelelectrophoresis (DGGE) technology was employed to reveal thestructure of the microbial community. Hence, this work could beuseful for the development of aerobic granulation systems and thewide application of ZVI in the field of biotechnology. To the best ofour knowledge, this is the first attempt to enhance aerobic granu-lation by adding ZVI.2. Materials and methods2.1. Experimental set-up and operationExperiments were performed in parallel using two double-walled, cylindrical column SBARs with internal diameter of 8 cm.The SBARs contained an internal riser (bottom clearance of 5 cm,internal diameter of 1.5 cm, and height of 80 cm) with a workingvolume of 3.6 L. Air was introduced via a fine bubble aerator througha porous stone diffuser at the bottom of the reactor at an airflowrate of 3 L per minute during the aeration phase.Both reactors were operated in successive cycles with each cyclelasting 6 h. One cycle consisted of five successive phases:(1)10 minfeeding,(2)310 min aerobic reaction,(3)10 min settling,(4)5 mindischarging
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