Comparison of primary and secondary sludge carbon sources derived from hydrolysis or acidogenesis for nitrate reduction and denitrification kinetics: Organics utilization and microbial community shift.

Seeking available and economical carbon sources for denitrification process is an intractable issue for wastewater treatment. However, no study compared different types of waste sludge as carbon source from denitrification mechanism, organics utilization and microbial community aspects. In this study, primary and secondary sludge were pretreated by thermophilic bacteria (TB), and its hydrolysis or acidogenic liquid were prepared as carbon sources for denitrification. At C/N of 8-3, the variations of NO3--N and NO2--N were profiled in typical cycles and denitrification kinetics was analyzed. Primary sludge achieved a competitive NOX-N removal efficiency with less dosage than secondary sludge. Fourier transform infrared (FTIR) spectroscopy was introduced to analyze organic composition from functional-group perspective and the utilization of organic matters in different sludge carbon sources was investigated. To further analyze the microbial community shift in different denitrification systems, high-throughput sequencing technology was applied. Results showed that denitrifier Thauera, belonging to Proteobacteria, was predominant, and primary sludge acidogenic liquid enriched Thauera most intensively with relative abundance of 47.3%.

Regulation of carbon source metabolism in mixotrophic microalgae cultivation in response to light intensity variation.

Microalgae are one of the promising sources for renewable energy production, and the light intensity variation can affect the biofuel generation and carbon assimilation of mixotrophic microalgae. To reveal the response of carbon assimilation to light intensity, the effect of light intensity on the carbon source metabolism of Chlorella vulgaris under mixotrophic cultivation was investigated in this study. Moreover, the optimal carbon source composition for mixotrophic microalgae cultivation was evaluated using bicarbonate (HCO3-) and carbonate (CO32-) as inorganic carbon sources, and glucose and acetate as organic carbon sources. The optimal light intensity for Chlorella vulgaris growth was at the range of 8000-12000 lux. For the accumulation of biochemical components, low light intensity was beneficial to protein accumulation, and high light intensity was advantageous for carbohydrate and lipid accumulation. With HCO3- and glucose, the maximum lipid content reached 37.0% at a light intensity of 12000 lux. The citrate synthase activity was negatively correlated with light intensity, showing an opposite trend to biomass production. High light intensity had a positive impact on Rubisco expression, which promoted the microalgae growth and carbon fixing. The energy produced by heterotrophic metabolic activities increased at low light intensity, and the enhancement of biomass production with high light intensity was mainly caused by the improved photoreaction efficiency during the mixotrophic cultivation.

Effect of external carbon addition and enrofloxacin on the denitrification and microbial community of sequencing batch membrane reactor treating synthetic mariculture wastewater.

The effect of sequencing batch membrane bioreactor (SMBR) on external carbon addition and enrofloxacin was investigated to treat synthetic mariculture wastewater. Anoxic/anaerobic and low COD/TN can improve the ammonia oxidation of the system, and the NH4+-N removal efficiency above 99%. External carbon was added and an anoxic environment was set to provide a suitable environment for denitrifying bacteria. When the external carbon source was 50-207 mg/L, the TN removal efficiency (31.82%-37.73%) and the COD of the effluent (28.85-36.58 mg/L) had little change. The partition resistance model showed that cake deposition resistance (RC,irr) and irreversible resistance (RPB) were the main components. And with the increase in cleaning times, the fouling rate of membrane components accelerated. Enrofloxacin can promote the TN removal efficiency (45.66%-93.74%) and had a significant effect on TM7a, Cohaesibacter, Vibrio and Phaeobacter.

Metagenomic analysis and nitrogen removal performance evaluation of activated sludge from a sequencing batch reactor under different salinities.

The effect of salinity on the nitrogen removal performance and microbial community of activated sludge was investigated in a sequencing batch reactor. The NH4+-N removal efficiency was over 95% at 0-4% salinity, indicating that the nitrification performance of activated sludge was slightly affected by lower salinity. The obvious nitrite accumulation was observed with the increment of the salinity to 5%, followed by a notable decline in the nitrogen removal performance at 6% salinity. The salinity inhibited the microbial activity, and the specific rate of nitrification and denitrification was decreased by the increasing salinity obviously. Additionally, the lower activity of superoxide dismutase and peroxidase and higher reactive oxygen species content in activated sludge might account for the deteriorative nitrogen removal performance at 6% salinity. Metagenomics analysis revealed that the genes encoding the ABC-type quaternary amine transporter in the ABC transporter pathway were abundant in the activated sludge at 2% and 4% salinity, and the higher salinity of 6% led to the loss of the genes encoding the p-type Na+ transporter in the ABC transporter pathway. These results indicated that the salinity could weaken the ABC transporter pathway for the balance of osmotic pressure in activated sludge. The microbial activity and nitrogen removal performance of activated sludge were decreased due to the unbalanced osmotic pressure at higher salinity.

Mariculture wastewater treatment with Bacterial-Algal Coupling System (BACS): Effect of light intensity on microalgal biomass production and nutrient removal.

Mariculture wastewater generated from the mariculture industry has increased public concern due to its impact on the sustainability of aquatic environments and aquaculture practices. Herein, the Bacterial-Algal Coupling System was applied for mariculture wastewater treatment. Microalgae growth in heterotrophy and mixotrophy (2000-8000 lux) was first compared. The best microalgal growth and nutrient removal were obtained at 5000 lux, where biomass productivity of microalgae was 0.465 g L-1 d-1, and 98.1% of chemical oxygen demand, 70.7% of ammonia-nitrogen, and 90.0% of total phosphorus were removed. To further understand the nutrient removal through microalgae cultivation, the enzyme activities involved in the Calvin cycle and the Tricarboxylic Acid cycle at different light intensities were determined. Under mixotrophic cultivation, there was a coordination between photosynthesis and heterotrophic metabolism in the agal cell, which resulted in a high algal biomass production and removal efficiency of nutrients. This study provided a novel insight into the bioremediation of mariculture wastewater and microalgae cultivation.

Impact of carbon/nitrogen ratio on the performance and microbial community of sequencing batch biofilm reactor treating synthetic mariculture wastewater.

The differences of cultured organism species, aquaculture model and supervisor mode lead to different carbon/nitrogen ratios in mariculture wastewater. Therefore, the performance, microbial community and enzymatic activity of sequencing batch biofilm reactor were compared in treating synthetic mariculture wastewater at different chemical oxygen demand/nitrogen (COD/N) ratios. Compared with COD/N ratio of 6, the ammonia-oxidizing rate and nitrite-oxidizing rate at COD/N ratio of 5, 4 and 3 increased by 3.66 % and 3.08 %, 11.19 % and 14.95 %, and 24.50 % and 32.54 %, respectively. Similarly, the ammonia monooxygenase and nitrite oxidoreductase activities increased by 3.50 % and 6.76 %, 11.09 % and 16.22 %, and 25.43 % and 39.19 % at COD/N ratio at 5, 4 and 3, respectively. However, the denitrifying rate and denitrification enzymatic activity declined with the decrease of C/N ratio from 6 to 3. The production, protein content and polysaccharide content of loosely bound extracellular polymeric substances (LB-EPS) and tightly bound EPS (TB-EPS) reduced with the decrease of COD/N ratio from 6 to 3. The abundance of nitrifying genera increased with the decrease of COD/N ratio from 6 to 3, whereas most of denitrification genera displayed a decreasing trend. The microbial co-occurrence pattern, keystone taxa and significant difference were altered with the decrease of COD/N ratio. Among the keystone taxa, Thauera, Denitromonas, Nitrosomonas and Denitratisoma had a close link with nitrogen transformation. The present results can provide some theoretical basis for evaluating the effect of carbon/nitrogen ratio on the nitrogen removal of biological wastewater treatment systems.

Comparison of the effects of salinity on microbial community structures and functions in sequencing batch reactors with and without carriers.

This study investigated and compared the microbial communities between a sequencing batch reactor (SBR) without carriers and a hybrid SBR with addition of carriers for the treatment of saline wastewater. The two systems were operated over 292 days with alternating aerobic/anoxic mode (temperature: 28℃, salinity: 0.0-3.0%). High removal efficiency of chemical oxygen demand (COD) and total inorganic nitrogen (TIN) was achieved in both the SBR (above 86.7 and 95.4% respectively) and hybrid SBR (above 84.4 and 94.0%) at 0.0-2.5% salinity. Further increasing salinity to 3.0% decreased TIN removal efficiency to 78.4% in the hybrid SBR. Steep decline of biodiversity and relative abundance of ammonia-oxidizing bacteria (AOB) contributed to the worse performance. More genera related to sulfide-oxidizing and sulfate-reducing bacteria were detected in the hybrid SBR than the SBR at 3.0% salinity. The abundance of halotolerant bacteria increased with the salinity increase for both reactors, summing up to 25.5% in the suspended sludge (S-sludge) from the SBR, 28.9 and 22.9% in the S-sludge and biofilm taken from the hybrid SBR, respectively. Nitrification and denitrification via nitrate was the main nitrogen removal pathway in the SBR and hybrid SBR at 0.0 and 0.5% salinity, while partial nitrification and denitrification via nitrite became the key process for nitrogen removal in the two reactors when the salinity was increased to 1.0-3.0%. Higher abundance of anaerobic ammonium-oxidizing (ANAMMOX) and sulfide-oxidizing autotrophic denitrification (SOAD) bacteria were found in the hybrid SBR at 3.0% salinity.

Effect of salinity and pH on dark fermentation with thermophilic bacteria pretreated swine wastewater.

The utilization of swine wastewater is affected by salinity and pH owing to the extensive use with seawater instead of domestic water as swine farm flushing water in coastal city. Therefore, swine wastewater pretreated with thermophilic bacteria was used as fermentation substrate in this work, the effects of salinity and pH on dark fermentation under mesophilic condition were investigated. The research showed that 1.5% salinity and pH 6.0 were the optimal conditions for hydrogen production with swine wastewater. The activity of hydrogenogen was inhibited at 3.5% salinity and pH 5.0. Soluble organic matter in substrate was accumulated under high salinity and alkaline conditions. The utilization of carbohydrate during dark fermentation was up to 61.1% at 1.5% salinity and 51.5% at pH 9.0. Enhancing of salinity and pH had an advantage in accumulation of total soluble metabolites. Acetate was the main metabolite during dark fermentation, and 1.5% salinity contributed to the formation of butyrate.

Effects of transient 3-chloroaniline shock loading on the performance, microbial community and enzymatic activity of sequencing batch reactor.

Chloroanilines from industrial wastewater can produce adverse effects on biological wastewater treatment systems due to their potential biotoxicity. The performance, nitrogen removal rate, microbial community and enzymatic activity of a sequencing batch reactor (SBR) were evaluated under transient 3-chloroaniline shock loading. After 40 mg/L 3-chloroaniline shock loading of 24 h on day 9, the chemical oxygen demand (COD) removal efficiency decreased from 90.71% on day 8 to 80.57% on day 11, and the NH4+-N removal efficiency reduced from 98.96% on day 8 to 35.51% on day 12. Subsequently, the COD and NH4+-N removal efficiencies gradually recovered to normal value. Compared with the absence of 3-chloroaniline shock loading, the ammonia-oxidizing rate (SAOR), nitrite-oxidizing rate (SNOR), nitrite-reducing rate (SNIRR) and nitrate-reducing rate (SNRR) decreased by 66.19%, 14.49%, 16.20% and 49.38% on day 11, respectively, and then they gradually recovered to normal value. The SAOR, SNOR, SNIRR and SNRR displayed the similar varying trends to the activities of ammonia monooxygenase, nitrite oxidoreductase, nitrite reductase and nitrate reductase, respectively. The appearance of 3-chloroaniline promoted the microbial reactive oxygen species production and lactate dehydrogenase release. The transient 3-chloroaniline shock loading distinctly impacted the microbial richness and diversity. The present research results can provide theoretical basis and technical support for evaluating the effects of transient 3-chloroaniline shock on biological wastewater treatment systems, which is beneficial to take reasonable preventable measures to decrease the adverse effects on the bioreactor performance.

Accelerating waste sludge hydrolysis with alkyl polyglucose pretreatment coupled with biological process of thermophilic bacteria: Hydrolytic enzyme activity and organic matters transformation.

A novel pretreatment method combining thermophilic bacteria (TB) with alkyl polyglucose (APG) was employed to pretreat waste sludge for enhancing the sludge hydrolysis. TB combined with APG pretreatment was effective in the releasing of soluble chemical oxygen demand (SCOD), protein and carbohydrate in extracellular polymeric substances (EPS) when the dosage of APG was below 0.1 g/g TSS. The enhancement of SCOD, carbohydrates and protein in dissolved organic matter (DOM) was promoted by the synthetic effect of APG and TB, which provides more carbon and energy source to the subsequent biochemical processes in sludge digestion. Excitation-emission matrix (EEM) fluorescence spectroscopy revealed that the combined pretreatment was beneficial for the decrease of non-biodegradable materials and the increase of biodegradable materials in DOM, resulting in the enhancement of the biodegradation of waste sludge. The combined use of TB and 0.4 g/g TSS APG achieved the maximal activities of protease (1.8) and α-glucosidase (1.9), and the activities of protease and α-glucosidase were positively correlated to the dosage of APG. The combined pretreatment was advantageous for the sludge reduction and sludge stabilization.

The influence of Fe2+, Fe3+ and magnet powder (Fe3O4) on aerobic granulation and their mechanisms.

This study aimed to develop an aerobic granular sludge and understand the granulation process of the multi-iron ions. Four sequencing batch reactors (SBRs) were applied to elucidate the effect of Fe2+, Fe3+ and Fe3O4 addition on aerobic granulation. The results confirmed that the start-up time of aerobic granulation with Fe3O4 addition (11 days) was notably less than that with Fe2+ (16 days) and Fe3+ (27 days) addition. Larger granules achieved with Fe3O4 addition with a sludge volume index (SVI30) of 28.50 mL/g and settling velocity of 49.68 m/h. Scanning electron microscope (SEM) analysis further revealed that the presence of mineral crystal in the granule core with Fe2+ and Fe3O4 addition accelerated the granule formation and maintained the stability of the structure. Extracellular polymeric substances (EPS) were studied using three-dimensional-excitation emission matrix (3D-EEM) fluorescence spectra technology to gain a comprehensive view of the interactions between EPS and Fe2+, Fe3+ and Fe3O4. Around 94.76% and 97.68% removal rate was noted for COD and ammonia in the granulation process. Finally, the dominant functional species involved in biological nutrients removal and granule formation were identified by high throughput sequencing technology to assess the effects of Fe2+, Fe3+ and Fe3O4 to granule at the molecular level.

Salinity effect on simultaneous nitrification and denitrification, microbial characteristics in a hybrid sequencing batch biofilm reactor.

The effect of increasing salinity on nitrogen removal via simultaneous nitrification and denitrification, microbial activities and extracellular polymeric substances (EPS) were investigated in a hybrid sequencing batch biofilm reactor filled with soft combination carriers. In the influent salinity range from 1.0 to 2.0%, average COD, NH4+-N and TN removal efficiencies were higher than 97.1, 97.8 and 86.4% at the steady state. When salinity was increased to 2.5 and 3.0%, ammonium oxidation was obviously inhibited in the reactor. For both suspended sludge (S-sludge) and biofilm, specific ammonium oxidation rate, specific nitrite oxidation rate, specific oxygen uptake rate and dehydrogenase activity reduced with the increase of salinity. The quantity of total EPS increased with the increase of salinity from 1.0 to 2.0%. Generally, humic substances were the dominant composition of EPS in both S-sludge and biofilm, with the percentages of 43.9-54.0 and 43.8-64.6% in total EPS.

Effect of magnesium oxide nanoparticles on microbial diversity and removal performance of sequencing batch reactor.

The performance, microbial enzymatic activity and microbial community of a sequencing batch reactor (SBR) have been explored under magnesium oxide nanoparticles (MgO NPs) stress. The NH4+-N removal efficiency kept relatively stable during the whole operational process. The MgO NPs at 30-60 mg/L slightly restrained the removal of chemical oxygen demand (COD), and the presence of MgO NPs also affected the denitrification and phosphorus removal. The specific oxygen uptake rate, nitrifying and denitrifying rates, phosphorus removal rate, and microbial enzymatic activities distinctly varied with the increase of MgO NPs concentration. The appearance of MgO NPs promoted more reactive oxygen species generation and lactate dehydrogenase leakage from activated sludge, suggesting that MgO NPs had obvious toxicity to activated sludge in the SBR. The protein and polysaccharide contents of extracellular polymeric substances from activated sludge increased with the increase of MgO NPs concentration. The microbial richness and diversity at different MgO NPs concentrations obviously varied at the phylum, class and genus levels due to the biological toxicity of MgO NPs.

Effect of Substrate Conversion on Performance of Microbial Fuel Cells and Anodic Microbial Communities.

Performance of microbial fuel cells (MFCs) was monitored during the influent nutrient change from lactate to glucose/acetate/propionate and then to lactate. Meanwhile, anodic microbial communities were characterized by culture-independent molecular biotechnologies. Results showed MFC performance recovered rapidly when the lactate was replaced by one of its metabolic intermediates acetate, while it needed a longer time to recover if lactate substrate was converted to glucose/propionate or acetate to lactate. Secondary lactate feed enhanced the enrichment of bacterial populations dominating in first lactate feed. Electricity-producing bacteria, Geobacter spp., and beneficial helpers, Anaeromusa spp. and Pseudomonas spp., revived from a low abundance as lactate secondary supply, but microbial communities were hard to achieve former profiles in structure and composition. Hence, microbial community profiles tended to recover when outside environmental condition were restored. Different substrates selected unique functional microbial populations.

Long-term effects of cupric oxide nanoparticles (CuO NPs) on the performance, microbial community and enzymatic activity of activated sludge in a sequencing batch reactor.

The long-term effects of cupric oxide nanoparticles (CuO NPs) on the performance, microbial activity and microbial community of activated sludge were investigated in a sequencing batch reactor (SBR). The SBR performance had no evident change at 0-10 mg/L CuO NPs, whereas the CuO NPs concentration at 30-60 mg/L affected the COD, NH4+-N and soluble orthophosphate (SOP) removal, nitrogen and phosphorus removal rate and microbial enzymatic activity of activated sludge. Some CuO NPs might be absorbed on the surface of activated sludge or penetrate the microbial cytomembrane into the microbial cell interior of activated sludge. Compared to 0 mg/L CuO NPs, the reactive oxygen species (ROS) production and lactate dehydrogenase (LDH) release increased by 43.6% and 56.4% at 60 mg/L CuO NPs, respectively. The variations of ROS production and LDH release demonstrated that CuO NPs could induce the toxicity towards the microorganisms and destroy the integrity of microbial cytomembrane in the activated sludge. High throughput sequencing of 16S rDNA indicated that CuO NPs could evidently impact on the microbial richness, diversity and composition of activated sludge in the SBR.

Impacts of silver nanoparticles on performance and microbial community and enzymatic activity of a sequencing batch reactor.

The performance, microbial community and enzymatic activity of a sequencing batch reactor (SBR) were evaluated under silver nanoparticles (Ag NPs) stress. Over 5 mg/L Ag NPs inhibited the COD and phosphorus removals, whereas the NH4+ removal kept stable during the whole operational period. The organic matter, nitrogen and phosphorus removal rates were obviously inhibited under Ag NPs stress, which showed similar varying trends with the corresponding microbial enzymatic activities. The change of Ag content in the activated sludge indicated that some Ag NPs were absorbed by the sludge. The presence of Ag NPs promoted the increase of reactive oxygen species (ROS) and lactate dehydrogenase (LDH) of microorganism due to the microbial response to the Ag NPs toxicity, which could impact on the microbial morphology and physiological functions. The presence of Ag NPs could produce some evident changes in the microbial community.

Characteristics of extracellular polymeric substances from sludge and biofilm in a simultaneous nitrification and denitrification system under high salinity stress.

The composition and distribution of extracellular polymeric substance (EPS) both from suspended sludge and attached biofilm were investigated in a simultaneous nitrification and denitrification (SND) system with the increase of the salinity from 1.0 to 3.0 %. Fourier-transform infrared (FTIR) spectroscopy and three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectroscopy were used to examine proteins (PN), polysaccharides (PS) and humic substances (HS) present in EPS. High total nitrogen removal (above 83.9 %) via SND was obtained in the salinity range of 1.0-2.5 %. Total EPS in the sludge increased from 150.2 to 200.6 mg/gVSS with the increase of salinity from 1.0 to 3.0 %, whereas the corresponding values in the biofilm achieved the maximum of 288.6 mg/g VSS at 2.0 % salinity. Dominant composition of EPS was detected as HS in both sludge and biofilm, having the percentages of 50.6-68.6 and 41.1-69.9 % in total EPS, respectively. Both PN and PS contents in soluble EPS (S-EPS), loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) of sludge and biofilm increased with the increased salinity. The FTIR spectrum and 3D-EEM fluorescence spectroscopy of S-EPS, LB-EPS and TB-EPS in the sludge and biofilm showed the changes of functional groups and conformations of the compositions in EPS with the increase of salinity. The results demonstrated that the characteristics of EPS varied from sludge to biofilm. The obtained results could provide a better understanding of the salinity effect on the EPS characteristics in a SND system.

Performance evaluation and microbial community of a sequencing batch biofilm reactor (SBBR) treating mariculture wastewater at different chlortetracycline concentrations.

The effects of chlortetracycline (CTC) on the performance, microbial activity, extracellular polymeric substances (EPS) and microbial community of a sequencing batch biofilm reactor (SBBR) were investigated in treating mariculture wastewater. Low CTC concentration (less than 6 mg/L) had no obvious effect on the SBBR performance, whereas high CTC concentration could inhibit the chemical oxygen demand (COD) and nitrogen removal of the SBBR. The microbial activity of the biofilm in the SBBR decreased with the increase of CTC concentration from 0 to 35 mg/L. The protein (PN) contents were always higher than the PS contents in both loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) at different CTC concentrations. The chemical compositions of LB-EPS and TB-EPS had obvious variations with the increase of CTC concentration from 0 to 35 mg/L. The high-throughput sequencing revealed the effects of CTC on the microbial communities of the biofilm at phylum, class and genus level. The relative abundances of some genera displayed a decreasing tendency with the increase of CTC concentration from 0 to 35 mg/L, such as Nitrospira, Paracoccus, Hyphomicrobium, Azospirillum. However, the relative abundances of the genera Flavobacterium, Aequorivita, Buchnera, Azonexus and Thioalbus increased with the increase of CTC concentration.

Comparison of multi-enzyme and thermophilic bacteria on the hydrolysis of mariculture organic waste (MOW).

Mariculture organic waste (MOW) is rich in organic matter, which is a potential energy resource for anaerobic digestion. In order to enhance the anaerobic fermentation, the MOW was hydrolyzed by multi-enzyme and thermophilic bacteria. It was advantageous for soluble chemical oxygen demand (SCOD) release at MOW concentrations of 6 and 10 g/L with multi-enzyme and thermophilic bacteria pretreatments. For multi-enzyme, the hydrolysis was not obvious at substrate concentrations of 1 and 3 g/L, and the protein and carbohydrate increased with hydrolysis time at substrate concentrations of 6 and 10 g/L. For thermophilic bacteria, the carbohydrate was first released at 2-4 h and then consumed, and the protein increased with hydrolysis time. The optimal enzyme hydrolysis for MOW was determined by measuring the changes of SCOD, protein, carbohydrate, ammonia and total phosphorus, and comparing with acid and alkaline pretreatments.

Optimization of VFAs and ethanol production with waste sludge used as the denitrification carbon source.

An acidification metabolite such as volatile fatty acids (VFAs) and ethanol could be used as denitrification carbon sources for solving the difficult problem of carbon source shortages and low nitrogen removal efficiency. A proper control of environmental factors could be essential for obtaining the optimal contents of VFAs and ethanol. In this study, suspended solids (SS), oxidation reduction potential (ORP) and shaking rate were chosen to investigate the interactive effects on VFAs and ethanol production with waste sludge. It was indicated that T-VFA yield could be enhanced at lower ORP and shaking rate. Changing the SS, ORP and shaking rate could influence the distribution of acetic, propionic, butyric, valeric acids and ethanol. The optimal conditions for VFAs and ethanol production used as a denitrification carbon source were predicted by analyzing response surface methodology (RSM).