Sulfate-reducing Up-flow Sludge Bed Research Articles

A novel biotechnology for enhanced ciprofloxacin removal via bioaugmentation of Paraclostridium sp.

To enhance the removal of ciprofloxacin (CIP, 1000–10,000 μg/L) in pharmaceutical wastewater, an efficient CIP-degrading bacterium, Paraclostridium sp. (strain S2), was immobilized and added into a sulfate-reducing up-flow sludge bed (SRUSB) bioreactor during long-term operation (over 80 days). CIP removal was increased by 18.7 % in the bioaugmented bioreactor with immobilized strain S2 compared to that in the control bioreactor under high CIP concentration (i.e., 10,000 μg/L), and the acute toxicity of the effluent in the bioaugmented bioreactor was significantly decreased by 81.1 % compared to that of the control. These results were attributed to the specific adaptability of strain S2 to environments via the alteration of its microbial physiology (e.g., extracellular polymeric substance (EPS) secretion, membrane permeability, cell surface charge, and hydrophobicity), which ensured its sustainability in the bioreactor, thus enhancing CIP removal. This study presents a novel biotechnology for CIP-contaminated wastewater treatment via the bioaugmentation by strain S2.

Insight into the Removal of Enoxacin in an Anaerobic Sulfur-Mediated Wastewater Treatment System: Performance, Kinetics and Mechanisms

The removal of enoxacin (ENO), a broad-spectrum fluoroquinolone antibiotic, was firstly examined in a sulfate-reducing up-flow sludge bed (SRUSB) bioreactor over a long-term operation (366 days). Over 94% of the ENO was removed in the SRUSB bioreactor via adsorption and biodegradation at different initial ENO concentrations (i.e., 25–1000 μg/L). Based on the results of the batch tests, the sulfate-reducing sludge exhibited a high ENO adsorption capacity within a kd of 22.7–28.9 L/g-SS. The adsorption of ENO by the sulfate-reducing sludge was a spontaneous (ΔG° < 0 KJ/mol) and exothermic (ΔH° < 0 KJ/mol) process including physisorption and chemisorption (absolute value of ΔH° = 51.882 KJ/mol). Moreover, ENO was effectively biodegraded by the sulfate-reducing sludge within specific rates of 2.5–161.3 μg/g-SS/d. The ENO biodegradation process in the sulfate-reducing sludge system was most accurately described by the first-order kinetic model. Collectively, our findings provide insight into the applicability of a sulfate-reducing sludge system for ENO-contaminated wastewater treatment.

Stress responses of sulfate-reducing bacteria sludge upon exposure to polyethylene microplastics

The stress responses of sulfate-reducing bacteria (SRB) sludge to polyethylene (PE) microplastic exposure were revealed for the first time. In this study, a lab-scale sulfate-reducing up-flow sludge bed reactor was continuously operated with different concentrations of PE microplastics in the feed (20, 100, and 500 microplastic particles (MPs)/L). Exposure to low levels of PE microplastics (i.e., 20 MPs/L) had a limited effect on SRB consortia, whereas higher levels of PE microplastics imposed apparent physiological stresses on SRB consortia. Despite this, the overall reactor performance, i.e., chemical oxygen demand removal and sulfate conversion, was less affected by prolonged exposure to PE microplastics. Moreover, as the concentration of PE microplastics increased, the SRB consortia promoted the production of extracellular polymeric substances to a greater extent, especially the secretion of proteins. As a result, protective effects against the cytotoxicity of PE microplastics were provided. Batch experiments further demonstrated that leaching additives from PE microplastics (including acetyl tri-n‑butyl citrate and bisphenol A, concentrations up to 5 μg/g sludge) exerted only a minor effect on the activity of SRB consortia. Additionally, microbial community analysis revealed active and potentially efficient sulfate reducers at different operational stages. Our results provide insight into the stress responses of SRB sludge under PE microplastic exposure and suggested that SRB consortia can gradually adapt to and resist high levels of PE microplastics. These findings may promote a better understanding of the stable operation of SRB sludge systems under specific environmental stimuli for practical applications.

Insights into the role of biochar on the acidogenic process and microbial pathways in a granular sulfate-reducing up-flow sludge bed reactor

In this study, the effect of biochar on sulfate reduction and anaerobic acidogenic process was explored in a granular sulfate-reducing up-flow sludge bed reactor in both long-term operation and batch tests. Both bioreactors had a high sulfate reduction efficiency of over 95% during the long-term operation, while the reactor with biochar addition showed higher sulfate reduction efficiency and stronger robustness against volatile fatty acids accumulation with a higher organic loading and sulfate loading rate. Batch tests showed that adding biochar significantly lessened the lag phase of the sulfate-reducing process, accelerated the adaption of acidogens, and facilitated both production and utilization of volatile fatty acids. The microbial pathways proved that biochar could regulate the acidification fermentation pathway and facilitate the enrichment of assimilative desulfurization bacteria. Overall, this study revealed that the acidogenic sulfate-reducing metabolic pathway could be enhanced by biochar, offering a potential application for effective sulfate-laden wastewater treatment.

Electrolysis-sulfate-reducing up-flow sludge bed-biological contact oxidation reactor for Norfloxacin removal from wastewater with high sulfate content

This study investigated the treatment of 100-mg/L Norfloxacin (NOR) wastewater containing high concentrations of sulfate through a combination of electrolysis, sulfate-reducing up-flow sludge bed (SRUSB), and biological contact oxidation reactor (BCOR) treatments. Results revealed that after 62 h, the reaction system had processed over 97% of the NOR. Additionally, electrolysis with sodium sulfate as the electrolyte transformed 87.8% of the NOR but only 33.5% of the total organic carbon (TOC). In the SRUSB, the TOC and SO42− contents were simultaneously reduced by 87.4% and 95.6%, respectively, providing a stable environment to the BCOR. In the BCOR, 36.3% and 85.9% of the NOR and TOC were degraded. High-performance liquid chromatography-tandem mass spectrometry analysis identified three possible degradation pathways under the attack of –OH during electrolysis, including defluorination, piperazinyl ring transformation, and quinolone ring transformation. Furthermore, the Illumina HiSeq sequencing results demonstrated that the sulfate-reducing bacteria (represented by Desulfobacter and Desulfobulbus) in the SRUSB and the sulfate-oxidizing bacteria (mainly consisting of Gammaproteobacteria and Alphaproteobacteria) in the BCOR played important roles in carbon chain oxidation and benzene ring opening and thoroughly degraded the electrolysis products. Thus, this method effectively overcomes the incomplete degradation and low removal efficiency issues associated with single electrolysis or biological methods in traditional processes.

Removal of sulfamethoxazole (SMX) in sulfate-reducing flocculent and granular sludge systems

This study investigated sulfamethoxazole (SMX) removal and fate in sulfate-reducing up-flow sludge bed (SRUSB) reactors inoculated with sulfate-reducing bacteria (SRB) granules and flocs. The resilience of SRB granules and flocs against varying pHs and hydraulic retention times (HRTs) was also examined. SRB granules and flocs efficiently removed SMX from wastewater, which was significantly higher than the aerobic sludge. SRB granules achieved significantly (p < 0.05) higher SMX removal (∼13.3 μg/g suspended solids (SS)-d) than the SRB flocs (∼11.2 μg/g SS-d) during 150-day of SRUSB reactors operation. The SMX removal by both granules and flocs was mainly attributed to biodegradation. Sorption also contributed to SMX removal, in which aromatic protein-like substances of extracellular polymeric substances played important role in SMX removal. In addition, SRB granules showed higher resilience than SRB flocs against varying pHs and HRTs. Thus, SRB-mediated biological process, especially SRB granules, could be a promising biotechnology to remove SMX from wastewaters.

Elucidating the effects of starvation and reactivation on anaerobic sulfidogenic granular sludge: Reactor performance and granular sludge transformation

In biological wastewater treatment, the bacteria starvation always challenges the stability of system operation. Yet, the effects of starvation and possibility of reactivation are less understood for anaerobic sulfidogenic system. Sulfidogenic systems use sulfate as electron acceptor for organic chemical oxygen demand (COD) degradation, so it will encounter two kind of starvations: (i) complete stoppage of wastewater flow (named complete food starvation) and (ii) remaining organic COD but with very low level sulfate in the influent (named sulfate starvation). In this study, the relative long-term starvation (over 30-day) and subsequent reactivation were studied in two lab-scale continuous gas recirculation sulfate-reducing upflow sludge bed (CGR-SRUSB) reactors. It was observed that the complete food starvation and sulfate starvation decreased the COD removal rate (in the similar range of 60–65%) and the specific sulfidogenic activity (about 45% and 61% respectively), as well as increasing the sludge flotation potential (SFP) from <15% to 58% and 35% respectively. Moreover, the following restoration experiments proved that the perturbed systems could be reactivated within 10–15 days for both cases. The results of investigating the mechanisms showed the performance deterioration were highly attributed to the starvation-induced granular sludge transitions, with respect to the changing of sludge physico-chemical properties (permeability, porosity, hydrophobicity and viscocity) and microbial stuctures (sulfate-reducting bacteria and extracellular polymeric substances). The outcomes of this study can provide useful information for dealing with the prolonged starvation problems in sulfidogenesis-based systems in industrial and municipal wastewater treatment.

Adsorption Mechanisms of Ciprofloxacin by Extracellular Polymeric Substances of Sulfate-reducing Bacteria Sludge

Extracellular polymeric substances (EPS) in microbial sludge, fulfils a key role in removal of micro-organic pollutants during biological wastewater treatment. In this study, the authors evaluated the removal of ciprofloxacin (CIP) by sulfate-reducing bacteria (SRB) sludge in a sulfate-reducing up-flow sludge bed (SRUSB) reactor, and examined the role of EPS on CIP removal in an SRB sludge system. The results indicated that CIP was removed efficiently through adsorption and biodegradation by SRB sludge, with adsorption the major removal pathway. EPS also played an important role in CIP adsorption by SRB sludge, and the adsorption mechanisms of CIP by EPS were investigated using the three-dimensional excitation-emission matrix fluorescence spectroscopy technologies combined with parallel factor analysis. The functional groups binding CIP onto EPS were identified through Fourier transform infrared (FTIR) spectra analysis. The results suggested that the static quenching of EPS following CIP adsorption led to formation of an EPS-CIP complex, and that the CIP was mainly bound with tryptophan and tyrosine-like protein substances in EPS with the binding constants of 1.43×104 L·mol-1 and 1.02×104 L·mol-1, respectively. The FTIR results suggested that hydroxyl, amino and carboxyl functional groups were mainly responsible for binding of CIP onto EPS. The results revealed the adsorption mechanisms of CIP by EPS in SRB sludge, and enhanced understanding of the role of EPS in sulfur-mediated biological processes for the removal of CIP and other organic micro-pollutants.

Characterization of a new continuous gas-mixing sulfidogenic anaerobic bioreactor: Hydrodynamics and sludge granulation

Continuous gas recirculation (CGR) was demonstrated in this study to be an effective method to mitigate the persistent problem of sludge flotation in high-rate sulfate-reducing upflow sludge bed (SRUSB) reactors that do not produce much gas. The effects of hydraulic- and CGR-mixing on the mixing regime of the SRUSB reactors were investigated over a period of 45 d at the average shear rates of 0.9, 1.5, 2.7, 4.2 and 7.2 s−1 (Phase I). CGR-mixing at 4.2 s−1 resulted in the smallest reactor short-circuiting flow of 1.3 ± 0.1% and the smallest dead zone volume of 0.2 ± 0.01% at a lower power consumption (0.0007 W) than hydraulic mixing. In Phase II, the SRUSB reactor with CGR-mixing at 4.2 s−1 was re-inoculated and operated for 150 days. Within the first 65 days, the sludge transformed into micro-granules (300–350 μm) with a high sulfate-reducing bacteria (SRB) activity (0.62 ± 0.05 g COD/(g MLVSS·day)), a low sludge flotation potential (<20%) and a high settleability (SVI5/SVI30 < 1.3). These results are attributed to the following sludge properties: (i) a low ratio of loosely-bound to tightly-bound extracellular polymeric substances (0.06–0.1), (ii) weakly adhesive surface properties as demonstrated by a strongly negative zeta potential (−23 ± 2 mV), a low hydrophobicity (37 ± 3%) and a low viscosity (0.7 ± 0.1 mPa s), and (iii) small size granules resulting in strong mass transfer (sulfate and COD penetration into the granule core) and a homogeneous structure (SRB detected throughout the granule).

Sulfamethoxazole degradation in anaerobic sulfate-reducing bacteria sludge system

Sulfamethoxazole (SMX) is one of the most commonly used antibiotics. SMX degradation in sulfate-reducing bacteria (SRB) sludge systems has not been reported so far. This research investigated the SMX degradation using SRB sludge in a sulfate-reducing up-flow sludge bed reactor. Moreover, the mechanisms and kinetics of SMX removal were also investigated using SRB sludge via a series of batch experiments. The results showed that SMX removal was characterized by a rapid sorption onto SRB sludge, and desorption from SRB sludge to aqueous phase until achieving equilibrium, and then followed by slow biodegradation. Biodegradation was the dominant route for SMX removal. The sorption process conformed well to a pseudo-second-order kinetic model, meaning that the sorption occurred primarily via a chemical sorption process. The removal of SMX followed the pseudo-zero-order kinetic model with a specific removal rate of 13.2 ± 0.1 μg/L/d at initial SMX concentration 100 μg/L in batch tests. Based on the analysis of metabolites, most of the SMX biotransformation products' structures altered in the isoxazole ring, which were significantly different from that produced by aerobic and anaerobic sludge systems. Thus, SRB sludge system could play an important role in SMX biodegradation, especially in Sulfate-reduction Autotrophic denitrification and Nitrification Integrated (SANI) process for sewage treatment.

A super high-rate sulfidogenic system for saline sewage treatment

This study proposes a novel approach to resolve the challenging issue of sludge bed clogging in a granular sulfate-reducing upflow sludge bed (GSRUSB) reactor by means of introducing intermittent gas sparging to advance it into a super high-rate anaerobic bioreactor. Over a 196-day lab-scale trial, the GSRUSB system was operated from nominal hydraulic retention time of 4-hr to 40-min and achieved the highest organic loading rate of 13.31 kg COD/m3·day which is substantially greater than the typical loading of 2.0–3.5 kg COD/m3·day in a conventional upflow anaerobic sludge bed reactor treating dilute organic strength wastewater. The average organic removal efficiency and total dissolved sulfide of this system were 90 ± 4.2% and 158 ± 28 mg S/L, while organics residual in the effluent was 34 ± 14 mg COD/L. The control stage (without gas sparging) revealed that the sludge bed clogging happened concomitantly with the significant drop in extracellular polymeric substance content of granular sludge, through relevant chemical measurements and confocal laser scanning microscopy analyses. On the other hand, compared with increasing the effluent recirculation ratio (from 1.4 to 5), the three-dimensional computational fluid dynamics modeling in combination with energy dissipation analysis demonstrated that the gas sparging (at a superficial gas velocity of 0.8 m s−1) can create a 23 times higher liquid shear as well as enhanced particle attrition. Overall, this study not only developed a super high-rate anaerobic bioreactor for saline sewage treatment, but also shed light on the role of intermittent gas sparging in control of sludge bed clogging for anaerobic bioreactors.

Physicochemical and biological characterization of long-term operated sulfate reducing granular sludge in the SANI® process

The SANI® process (Sulfate reduction, Autotrophic denitrification and Nitrification Integrated) is a treatment system with low energy demands. The major bioreactor of this new technology is a sulfate-reducing up-flow sludge bed (SRUSB) that converts organics and provides electron donors for subsequent autotrophic denitrification. This research characterizes the granules inside the SRUSB, with the aim of improving its efficiency, maximizing its operational flexibility, and minimizing its footprint. The unique sulfate-reducing bacteria (SRB) granules serving in the SRUSB were found to increase the resilience and compactness of the SRUSB. The granules, with a compact and porous structure, showed high cohesion resisting breakage with a shear force G > 3400 s−1. The hydrophobicity of the external surface of the mature granules remained stable at around 70% and acid volatile sulfide (AVS) accumulated at the bottom of the SRUSB. 16s rRNA gene analysis of the microbial communities revealed that Desulfobulbus (42.1%), Prosthecochloris (19%) and Trichococcus (12%) dominated the mature granular sludge. Fluorescence in situ hybridization (FISH) further showed that SRB organisms were located internally and then surrounded by non-SRB. According to the FISH results, the spatial distribution of extracellular polymeric substances (EPS) displayed protein and α-polysaccharides in the exterior and β-polysaccharide in the core of the granules. Such biological structure suggests that each SRB granule acts as an efficient and independent unit, capable of achieving both fermentation and organic conversion. The present investigation sheds light on the physicochemical and biological characteristics of the SRB granulate. This information provides valuable information for scaling-up the SANI® process to treat real saline sewage in Hong Kong.

Characterization of sulfate-reducing granular sludge in the SANI® process

Hong Kong practices seawater toilet flushing covering 80% of the population. A sulfur cycle-based biological nitrogen removal process, the Sulfate reduction, Autotrophic denitrification and Nitrification Integrated (SANI®) process, had been developed to close the loop between the hybrid water supply and saline sewage treatment. To enhance this novel process, granulation of a Sulfate-Reducing Up-flow Sludge Bed (SRUSB) reactor has recently been conducted for organic removal and provision of electron donors (sulfide) for subsequent autotrophic denitrification, with a view to minimizing footprint and maximizing operation resilience. This further study was focused on the biological and physicochemical characteristics of the granular sulfate-reducing sludge. A lab-scale SRUSB reactor seeded with anaerobic digester sludge was operated with synthetic saline sewage for 368 days. At 1 h nominal hydraulic retention time (HRT) and 6.4 kg COD/m3-d organic loading rate, the SRUSB reactor achieved 90% COD and 75% sulfate removal efficiencies. Granular sludge was observed within 30 days, and became stable after 4 months of operation with diameters of 400–500 μm, SVI5 of 30 ml/g, and extracellular polymeric substances of 23 mg carbohydrate/g VSS. Fluorescence in situ hybridization (FISH) analysis revealed that the granules were enriched with abundant sulfate-reducing bacteria (SRB) as compared with the seeding sludge. Pyrosequencing analysis of the 16S rRNA gene in the sulfate-reducing granules on day 90 indicated that the microbial community consisted of a diverse SRB genera, namely Desulfobulbus (18.1%), Desulfobacter (13.6%), Desulfomicrobium (5.6%), Desulfosarcina (0.73%) and Desulfovibrio (0.6%), accounting for 38.6% of total operational taxonomic units at genera level, with no methanogens detected. The microbial population and physicochemical properties of the granules well explained the excellent performance of the granular SRUSB reactor.

Granulation of anaerobic sludge in the sulfate-reducing up-flow sludge bed (SRUSB) of SANI® process

This study reports on anaerobic sludge granulation in a laboratory-scale sulfate-reducing up-flow sludge bed (SRUSB) in a novel sulfate reduction, autotrophic denitrification and nitrification integrated (SANI(®)) process for treatment of saline sewage. Granulation occurred in 30 d and reached full development in 90 d. The sulfate-reducing granules grew up to around 1 mm after 90 d with 21 mL/g SVI5 (sludge volume index measured after 5 min) and the biomass concentration reached 29 g/L after 4 months' operation. The reactor removed 89% chemical oxygen demand (COD) and reduced 75% sulfate within 1 h of hydraulic retention time, under a COD loading rate of up to 6.4 kg COD/(m(3) · d).

Microbial community of sulfate-reducing up-flow sludge bed in the SANI® process for saline sewage treatment

This study investigated the microbial community of the sulfate-reducing up-flow sludge bed (SRUSB) of a novel sulfate reduction, autotrophic denitrification, and nitrification integrated (SANI®) process for saline sewage treatment. The investigation involved a lab-scale SANI® system treating synthetic saline sewage and a pilot-scale SANI® plant treating 10m(3)/day of screened saline sewage. Sulfate-reducing bacteria (SRB) were the dominant population, responsible for more than 80% of the chemical oxygen demand removal, and no methane-producing archaea were detected in both SRUSBs. Thermotogales-like bacteria were the dominant SRB in the pilot-scale SRUSB while Desulforhopalus-like bacteria were the major species in the lab-scale SRUSB.

Fecal coliform removal in a sulfate reduction, autotrophic denitrification and nitrification integrated (SANI) process for saline sewage treatment

The Sulfate Reduction, Autotrophic Denitrification, Nitrification Integrated (SANI) process has been specially designed to treat saline wastewater. In the process no biological excess sludge is produced. SANI process also has the added advantages of cost and footprint reduction when compared to traditional activated sludge processes. In the SANI pilot plant, the fecal coliform removal efficiency in the sulfate reducing up-flow sludge bed (SRUSB) was found to be 1.4 log, whereas that in the subsequent anoxic and aerobic reactors it was 0.6 and 0.2 log, respectively, leading to a relatively high overall coliform removal of 2.2 log. Sulfide was confirmed to be toxic to fecal coliform and the contact time between the sulfide produced and coliform in the SRUSB played an important role in the removal.