Nanobubble technology to enhance energy recovery from anaerobic digestion of organic solid wastes: Potential mechanisms and recent advancements.

Nanobubble (NB) technology has gained popularity in the environmental field owing to its distinctive characteristics and ecological safety. More recently, the application of NB technology in anaerobic digestion (AD) systems has been proven to promote substrate degradation and boost the production of biogas (H2 and/or CH4). This review presents the recent advancements in the application of NB technology in AD systems. Meanwhile, it also sheds light on the underlying mechanisms of NB technology that contribute to the enhanced biogas production from AD of organic solid wastes. Specifically, the working principles of the NB generator are first summarized, and then the structure of the NB generator is optimized to accommodate the demand for NB characteristics in the AD system. Subsequently, it delves into a detailed discussion of how the addition of nanobubble water (NBW) affects AD performance and the different factors that NB can potentially contribute. As a simple and environmentally friendly additive, NBW was commonly used in the AD process to enhance the fluidity and mass transfer characteristics of digestate. Additionally, NB has the potential to enhance the functionality of different types of microbial enzymes that play crucial roles in the AD process. This includes boosting extracellular hydrolase activities, optimizing coenzyme F420, and improving cellulase function. Finally, it is proposed that NBW has development potential for the pretreatment of substrate and inoculum, with future development being directed towards this aim.

Comparing the effects of magnetite-mediated direct interspecies electron transfer with biogas mixing-driven interspecies hydrogen transfer on anaerobic digestion.

Although the promotive effect of direct interspecies electron transfer (DIET) on methane production has been well-documented, the practical applicability of DIET in different scenarios have not yet been systematically studied. This study compared the effects of magnetite-mediated DIET with conventional biogas mixing-driven interspecies hydrogen transfer (IHT) on anaerobic digestion (AD) of swine manure (SM). Compared with control, magnetite supplementation, biogas circulation, and their integration enhanced the CH4 yield by 19.3%, 25.9%, and 26.2%, respectively. Magnetite mainly enriched DIET-related syntrophic bacteria (Anaerolineae and Synergistia) and methanogens (Methanosarcina) to accelerate acidification and establish DIET, while biogas circulation mainly enriched hydrolytic bacteria (Clostridia) and hydrogenotrophic methanogens (Methanolinea and Methanobacterium) to promote hydrolysis and accelerate IHT. Coupling magnetite addition with biogas circulation led to the enrichment of the above six microorganisms to different extents. The effectiveness of the strategies for lowering the H2 pressure followed: magnetite + biogas circulation ≈ biogas circulation > magnetite. Under stress-free environment, the enhancement effect of magnetite-induced DIET was not even as pronounced as biogas circulation-a simple and common mixing strategy in commercial AD plants, and the promotion effect of magnetite was insignificant in the well-mixed digesters. In short, the magnetite-mediated DIET is not always effective in improving AD of SM.

Reduction of greenhouse gas emissions from closed activated sludge- to aerobic granular sludge-based biosystems via gas circulation.

Greenhouse gas (GHG) emissions from biological treatment units are challenging wastewater treatment plants (WWTPs) due to their wide applications and global warming. This study aimed to reduce GHG emissions (especially N2O) using a gas circulation strategy in a closed sequencing-batch reactor when the biological unit varies from activated sludge (AS) to aerobic granular sludge (AGS). Results show that gas circulation lowers pH to 6.3 ± 0.2, facilitating regular granules but elevating total N2O production. From AS to AGS, N2O emission factor increased (0.07-0.86 %) due to decreasing ammonia-oxidizing rates while the emissions of CO2 (0.3 ± 0.1 kg-CO2/kg-chemical oxygen demand) and CH4 remained in the closed biosystem. The gas circulation decreased N2O emission factor by 63 ± 15 % after granulation higher than 44 ± 34 % before granulation, which is implemented by heterotrophic denitrification. This study provides a feasible strategy to enhance heterotrophic N2O elimination in the biological WWTPs.

Unveiling the role of ferrous ion in driving microalgae granulation from salt-tolerant strains for mariculture wastewater treatment.

Development of microalgal-bacterial granular sludge (MBGS) from saline-adapted microalgae is a promising approach for efficient mariculture wastewater treatment, whereas the elusive mechanisms governing granulation have impeded its widespread adoption. In this study, spherical and regular MBGS were successfully developed from mixed culture of pure Spirulina platensis and Chlorella sp. GY-H4 at 10 mg/L Fe2+ concentration. The addition of Fe2+ was proven to induce the formation of Fe-precipitates which served as nucleation sites for microbial attachment and granulation initiation. Additionally, Fe2+ increased the prevalence of exopolysaccharide-producing cyanobacteria, i.e. Synechocystis and Leptolyngbya, facilitating microbial cell adhesion. Furthermore, it stimulated the secretion of extracellular proteins (particularly tryptophan and aromatic proteins), which acted as structural backbone for the development of spherical granule form microalgal flocs. Lastly, it fostered the accumulation of exogenous heterotrophic functional genera, resulting in the efficient removal of DOC (98 %), PO43--P (98 %) and NH4+-N (87 %). Nevertheless, inadequate Fe2+ hindered microalgal floc transformation into granules, excessive Fe2+ expanded the anaerobic zone within the granules, almost halved protein content in the TB-EPS, and inhibited the functional genes expression, ultimately leading to an irregular granular morphology and diminished nutrient removal. This research provides valuable insights into the mechanisms by which Fe2+ promotes the granulation of salt-tolerant microalgae, offering guidance for the establishment and stable operation of MBGS systems in mariculture wastewater treatment.

MlrA, an Essential Enzyme for Microcystins and Nodularin on First Step Biodegradation in Microcystin-Degrading Bacteria.

Microcystin-degrading bacteria first degrade microcystins by microcystinase A (MlrA) to cleave the cyclic structure of microcystins at the Adda-Arg site of microcystin-LR, microcystin-RR, and microcystin-YR, but the cleavage of the other microcystins was not clear. In our study, the microcystin-degrading bacterium Sphingopyxis sp. C-1 as wild type and that of mlrA-disrupting mutant, Sphingopyxis sp. CMS01 were used for microcystins biodegradation. The results showed MlrA degraded microcystin-LA, microcystin-LW, microcystin-LY, microcystin-LF, and nodularin. MlrA could cleave the Adda-L-amino acid site.

Unveiling the mechanisms of Fe(III)-loaded chitosan composite (CTS-Fe) in enhancing anaerobic digestion of waste activated sludge.

Anaerobic digestion (AD) of waste activated sludge (WAS) is usually limited by the low generation efficiency of methane. Fe(III)-loaded chitosan composite (CTS-Fe) have been reported to effectively enhanced the digestion of WAS, but its role in promoting anaerobic sludge digestion remains unclear. In present study, the effects of CTS-Fe on the hydrolysis and methanogenesis stages of WAS anaerobic digestion were investigated. The addition of CTS-Fe increased methane production potential by 8%-23% under the tested conditions with the addition of 5-20 g/L CTS-Fe. Besides, the results demonstrate that the addition of CTS-Fe could effectively promote the hydrolysis of WAS, evidenced by lower protein or polysaccharides concentration, higher soluble organic carbon in rector adding CTS-Fe, as well as the increased activity of extracellular hydrolase with higher CTS-Fe concentration. Meanwhile, the enrichment of Clostridia abundance (iron-reducing bacteria (IRBs)) was observed in CTS-Fe adding reactor (8.9%-13.8%), which was higher than that in the control reactor (7.9%). The observation further suggesting the acceleration of hydrolysis through dissimilatory iron reduction (DIR) process, thus providing abundant substrates for methanogenesis. However, the presence of CTS-Fe was inhibited the acetoclastic and hydrogenotrophic methanogenesis process, which could be ascribed to the Fe(III) act as electron acceptor coupled to methane for anaerobic oxidation. Furthermore, coenzyme F420 activity in the CTS-Fe added reactor was 34.9% lower than in the blank, also abundance of microorganisms involved in hydrogenotrophic methanogenesis was decreased. Results from this study could provide theoretical support for the practical applications of CTS-Fe.

Photosynthetic oxygen-supported algal-bacterial aerobic granular sludge can facilitate carbon, nitrogen and phosphorus removal from wastewater: Focus on light intensity selection.

Photosynthetic O2 is a promising alternative for mechanical aeration, the major energy-intensive unit in wastewater treatment plants. This study aimed to investigate the effects of light intensity varied from 190 to 1400 µmol·s-1·m-2 on photosynthetic O2-supported algal-bacterial aerobic granular sludge (AGS) system. Results indicate photosynthetic O2 can implement aerobic phosphorus (P) uptake and ammonia oxidation under the test illumination range even at dissolved oxygen concentration < 0.5 mg/L. An obvious O2 accumulation occurred after 60-90% nutrients being removed under 330-1400 µmol·s-1·m-2, and highly efficient ammonia removal, P uptake, and dissolved inorganic carbon removal were achieved under 670-1400 µmol·s-1·m-2. On the other hand, photosynthesis as O2 supplier showed little effect on major ions except for K+. This study provides a better understanding of the roles of light intensity on photosynthetic O2-supported algal-bacterial AGS system, targeting a sustainable wastewater industry.

Coupling Iron Sludge Addition and Intermittent Aeration for Achieving Simultaneous Methanogenesis, Feammox, and Denitrification in a Single Reactor Treating Fish Sludge.

An integrated anaerobic digestion system for the simultaneous removal of carbon and nitrogen from fish sludge was developed by coupling iron sludge supplementation with intermittent aeration. In terms of nitrogen removal, Fe(III) in iron sludge could trigger Feammox reactions and intermittent aeration could drive the Fe(II)/Fe(III) cycle to sustain continuous ammonia removal. Mass balance analysis suggested that nitrate was the main product of Feammox, which was subsequently removed through heterotrophic denitrification. In terms of carbon removal, the Fe(III)-induced dissimilatory iron reduction (DIR) process significantly promoted fish sludge hydrolysis and provided more simple organics for methanogens and denitrifiers, but aeration showed a negative impact on methanogenesis. To promote nitrogen removal and avoid serious methanogenesis inhibition, different aeration intensities were studied. Results showed that compared with the control without aeration or iron sludge addition, aeration for 5 min every 3 days (150 mL/min) contributed to a 29.0% lower NH4+-N concentration and a 12.1% lower total chemical oxygen demand level on day 28, and the decline in methane yield was acceptable (only 13.5% lower). Simultaneous methanogenesis, Feammox, and denitrification in a single reactor treating fish sludge were achieved, which provides a simple and low-cost strategy for the treatment of organic wastewater.

Response of viable bacteria to antibiotics in aerobic granular sludge: Resistance mechanisms and behaviors, bacterial communities, and driving factors.

The assessment of antimicrobial resistance (AMR) risk by DNA-based techniques mainly relies on total bacterial DNA. In this case, AMR risk recognition is restricted to the genotype level, lacking crucial phenotypic information, such as the distribution of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in dead and viable bacteria. This limitation hinders the recognition of AMR behavior. Herein, based on propidium monoazide (PMA) shielding method, this work firstly quantified the intracellular ARGs/MGEs in viable and dead bacteria, and the impact of viable bacteria composition on the formation of intracellular/extracellular polymeric substance-related /cell-free ARGs (i/e/cARGs) and MGEs (i/e/cMGEs) in aerobic granular sludge (AGS). The shielding efficiency of PMA against dead bacteria was optimized to be as high as 97.5% when the MLSS of AGS was 2.0 g/L. Under antibiotic stimulation, 29.0% ∼ 49.0% of iARGs/iMGEs were carried by viable bacteria, and the remaining proportion were carried by dead bacteria. 18 out of the top 20 dominant genera showed a change in abundance by more than 1% after PMA treatment. 29 viable hosts were identified to associate with 52 iARGs, of which 28 and 15 hosts were also linked to 40 eARGs and 26 cARGs. Also, partial least-squares path model and variance partitioning analysis disclosed that viable bacteria and i/e/cMGEs had a positive effect on i/e/cARGs, with both contributing as much as 64.5% to the total ARGs enrichment. These results better visualized the AMR risk carried by viable bacteria and the categories of viable hosts. This work provides a novel insight into analyzing the actual AMR risk and viable hosts, helping to the reduction and control of AMR in wastewater treatment plants.

Enhanced alginate-like exopolymers recovery from algal-bacterial aerobic granular sludge: Optimal cultivation condition and contribution of bacteria and microalgae during the transport/storage period.

Extracting alginate-like exopolymers (ALE) is a promising approach for valuable resources recovery from excess algal-bacterial aerobic granular sludge (AGS) to achieve circular bioeconomy and environmental sustainability in wastewater treatment plants (WWTPs). In this study, six batch cultivation tests were conducted to investigate the optimal cultivation duration or transport/storage period, light intensity, and temperature for algal-bacterial AGS after sampling and before further processing or ALE extraction. At a light intensity of 5 klux, the highest ALE content (36.33 mg/g-VSS) was detected at a low temperature of 10 °C, which increased by 300% from its original level after 6-h cultivation. Results from levofloxacin (LVX) exposure and dark condition imply that microalgae contributed more to ALE synthesis in the algal-bacterial granules. This work not only provides a better understanding of the mechanisms involved in ALE biosynthesis but also offers useful guidance for maintaining or improving ALE recovery after algal-bacterial biomass sampling.

Highly efficient carbon assimilation and nitrogen/phosphorus removal facilitated by photosynthetic O2 from algal-bacterial aerobic granular sludge under controlled DO/pH operation.

Reducing CO2 emission and energy consumption is crucial for the sustainable management of wastewater treatment plants (WWTPs). In this study, an algal-bacterial aerobic granular sludge (AGS) system was developed for efficient carbon (C) assimilation and nitrogen (N)/phosphorus (P) removal without the need for mechanical aeration. The photosynthetic O2 production by phototrophic organisms maintained the dissolved oxygen (DO) level at 3-4 mg/L in the bulk liquid, and an LED light control system reduced 10-30% of light energy consumption. Results showed that the biomass assimilated 52% of input dissolved total carbon (DTC), and the produced O2 simultaneously facilitated aerobic nitrification and P uptake with the coexisting phototrophs serving as a C fixer and O2 supplier. This resulted in a stably high total N removal of 81 ± 7% and an N assimilation rate of 7.55 mg/(g-MLVSS∙d) with enhanced microbial assimilation and simultaneous nitrification/denitrification. Good P removal of 92-98% was maintained during the test period at a molar ∆P/∆C ratio of 0.36 ± 0.03 and high P release and uptake rates of 10.84 ± 0.41 and 7.18 ± 0.24 mg/(g- MLVSS∙h), respectively. Photosynthetic O2 was more advantageous for N and P removal than mechanical aeration. This proposed system can contribute to a better design and sustainable operation of WWTPs using algal-bacterial AGS.

Characteristics of algal-bacterial aerobic granular sludge treating real wastewater: Effects of algal inoculation and alginate-like exopolymers recovery.

Limited information is available on the characteristics of algal-bacterial aerobic granular sludge (AGS) treating real wastewater, especially on its alginate-like exopolymers (ALE) production. In addition, the effect of target microalgae species inoculation on the system performance has not been fully understood. This study aimed to reveal the effect of microalgae inoculation on the characteristics of algal-bacterial AGS and its ALE production potential. Two photo-sequencing batch reactors (PSBR) were employed, namely R1 with activated sludge and R2 with Tetradesmus sp. and activated sludge being inoculated, respectively. Both reactors were fed with locally sourced municipal wastewater and operated for 90 days. Algal-bacterial AGS were successfully cultivated in both reactors. No significant difference was observed between the performances of R1 and R2, reflecting that the inoculation of target microalgae species may not be crucial for the development of algal-bacterial AGS when treating real wastewater. Both reactors achieved an ALE yield of about 70 mg/g of volatile suspended solids (VSS), indicating that a substantial amount of biopolymer can be recovered from wastewater. Interestingly, boron was detected in all the ALE samples, which might contribute to granulation and interspecies quorum sensing. The enrichment of lipids content in ALE from algal-bacterial AGS treating real wastewater reveals its high resource recovery potential. Overall, the algal-bacterial AGS system is a promising biotechnology for simultaneous municipal wastewater treatment and resource (like ALE) recovery.

Field ponding water exacerbates the dissemination of manure-derived antibiotic resistance genes from paddy soil to surrounding waterbodies.

Farmlands fertilized with livestock manure-derived amendments have become a hot topic in the dissemination of antibiotic resistance genes (ARGs). Field ponding water connects rice paddies with surrounding water bodies, such as reservoirs, rivers, and lakes. However, there is a knowledge gap in understanding whether and how manure-borne ARGs can be transferred from paddy soil into field ponding water. Our studies suggest that the manure-derived ARGs aadA1, bla1, catA1, cmlA1-01, cmx(A), ermB, mepA and tetPB-01 can easily be transferred into field ponding water from paddy soil. The bacterial phyla Crenarchaeota, Verrucomicrobia, Cyanobacteria, Choloroflexi, Acidobacteria, Firmicutes, Bacteroidetes, and Actinobacteria are potential hosts of ARGs. Opportunistic pathogens detected in both paddy soil and field ponding water showed robust correlations with ARGs. Network co-occurrence analysis showed that mobile genetic elements (MGEs) were strongly correlated with ARGs. Our findings highlight that manure-borne ARGs and antibiotic-resistant bacteria in paddy fields can conveniently disseminate to the surrounding waterbodies through field ponding water, posing a threat to public health. This study provides a new perspective for comprehensively assessing the risk posed by ARGs in paddy ecosystems.

The role of light wavelengths in regulating algal-bacterial granules formation, protein and lipid accumulation, and microbial functions.

High value-added products recovery from algal-bacterial granular sludge (ABGS) has received great attention recently. This study aimed to explore the role of different light wavelengths in regulating granule formation, protein and lipid production, and microbial functions. Bacterial granular sludge (BGS, R0) was most conducive to forming ABGS under blue (R2) light with the highest chlorophyll a (10.2 mg/g-VSS) and diameter (1800 µm), followed by red (R1) and white (R3) lights. R0-R3 acquired high protein contents (>164.8 mg/g-VSS) with essential amino acids above 44.4%, all of which were suitable for recycling, but R2 was the best. Also, blue light significantly increased total lipid production, while red light promoted the accumulation of some unsaturated fatty acids (C18:2 and C18:3). Some unique algae and dominant bacteria (e.g., Stigeoclonium, Chlamydomonas, and Flavobacteria) enrichment and some key functions (e.g., amino acid, fatty acid, and lipid biosynthesis) up-regulation in R2 might help to improve proteins and lipids quality. Combined, this study provides valuable guidance for protein and lipid recovery from ABGS.

Enhanced fixation of dissolved inorganic carbon by algal-bacterial aerobic granular sludge during treatment of low-organic-content wastewater.

The microalgae-based wastewater treatment technologies are believed to contribute to carbon neutrality. This study investigated the inorganic carbon fixation performance in the algal-bacterial aerobic granular sludge (A-BAGS) process under cultivation at different concentrations of organic carbon (OC) and inorganic carbon (IC). The results indicated that A-BAGS in treating wastewater containing organics of 77 mg-C/L contributed little to the fixation of inorganic carbon, while the highest inorganic carbon removal efficiency of 50 % was achieved at the influent IC of 100 mg/L and OC of 7 mg/L. This high IC condition contributed to enhanced biomass growth rate and enhanced extracellular polymeric substances, while it did not affect the granular stability and nitrification efficiency. The microbial diversity was also largely enhanced. The results demonstrated the great potential of A-BAGS for simultaneous resource recovery in wastewater and inorganic carbon fixation, while operation conditions need to be further optimized.

Biological nitrogen removal mechanisms during anaerobic digestion of swine manure: Effects of biogas circulation and activated carbon addition.

This study investigated the biological nitrogen removal mechanisms during the anaerobic digestion of swine manure and the effects of biogas circulation and activated carbon (AC) addition. Biogas circulation, AC addition, and their combination increased the methane yield by 25.9%, 22.3%, and 44.1%, respectively, when compared to the control. Nitrogen species analysis and metagenomic results indicated that nitrification-denitrification was the dominant ammonia removal pathway in all digesters with little oxygen, while anammox did not occur. Biogas circulation could promote mass transfer and induce air infiltration to enrich nitrification- and denitrification-related bacteria and functional genes. And AC might act as an electron shuttle to facilitate ammonia removal. The combined strategies showed a synergetic effect on the enrichment of nitrification and denitrification bacteria and functional genes, significantly lowering the total ammonia nitrogen by 23.6%. A single digester with biogas circulation and AC addition could enhance methanogenesis and ammonia removal via nitrification and denitrification.

Algal-bacterial aerobic granular sludge for real municipal wastewater treatment: Performance, microbial community change and feasibility of lipid recovery.

Despite various research works on algal-bacterial aerobic granular sludge for wastewater treatment and resource recovery processes, limited information is available on its application in real wastewater treatment in terms of performance, microbial community variation and resource recovery. This study investigated the performance of algal-bacterial aerobic granular sludge on real low-strength wastewater treatment in addition to the characterization of microbial community and fatty acid compositions for biodiesel production. The results demonstrated 71% COD, 77% NH4+-N and 31% phosphate removal efficiencies, respectively. In addition, all the water parameters successfully met the effluent standard A, imposed by the Department of Environment (DOE) Malaysia. Core microbiome analyses revealed important microbial groups (i.e., Haliangium ochraceum, Burkholderiales and Chitinophagaceae) in bacterial community. Meanwhile the photosynthetic microorganisms, such as Oxyphotobacteria and Trebouxiophyceae dominated the algal-bacterial aerobic granular sludge, suggesting their important roles in granulation and wastewater treatment. Up to 12.51 mg/gSS lipid content was recovered from the granules. In addition, fatty acids composition showed high percetages of C16:0 and C18:0, demonstrating high feasibility to be used for biodiesel production application indicated by the cetane number, iodine value and oxidation stability properties.

Use of nanobubble water bioaugmented anaerobically digested sludge for high-efficacy energy production from high-solids anaerobic digestion of corn straw.

An increasing attention has been paid to the secure and sustainable management of agricultural wastes, especially lignocellulosic biomass. Nanobubble water (NBW) contains 106-108 bubbles/mL with diameter <1000 nm. Although previous studies have examined the enhancement effects of NBW on methane production from organic solid wastes, the NBW-based anaerobic digestion (AD) system is still restrained from practical application due to the large increase in AD reactor volume, generation of wastewater, and increase in energy consumption as well. In this study, NBW bioaugmentation of anaerobically digested sludge for the first time was performed for high-solids AD of corn straw. Results show that cellulase, xylanases and lignin peroxidase activities were increased by 2-55% during the NBW bioaugmentation process. Significant enrichment of hydrolytic/acidogenic bacteria and methanogenic archaea were noticed in the NBW bioaugmented sludge. This study clearly demonstrated 47% increase in methane production from high-solids AD of corn straw when O2-NBW bioaugmented sludge was applied, achieving a net energy gain of 5138 MJ/t-volatile solids of corn straw with an energy recovery of 34%. The NBW-based high-solids AD system can provide a novel and sustainable management solution for renewable energy production from agricultural wastes, targeting the reduction of environmental pollution and energy crisis.

Pb(II) bioremediation using fresh algal-bacterial aerobic granular sludge and its underlying mechanisms highlighting the role of extracellular polymeric substances.

Lead (Pb) discharged from rural industries poses a significant threat to the environment and human health. Algal-bacterial aerobic granular sludge (A-B AGS) is a promising alternative for sewage treatment with high efficiency and good settleability. In this study, Pb(II) biosorption using fresh A-B AGS was investigated for the first time. The important role of extracellular polymeric substances (EPS) was revealed with the involved mechanisms being clarified. The desorbents for Pb recovery from Pb-loaded A-B AGS were also screened. Results showed that A-B AGS has an excellent maximum Pb adsorption capacity of 72.4 mg·g-1 at pH 6.0. EPS plays an important role in keeping microbial activity, Pb bonding, and providing metal ions (Ca, Na and Mg) for Pb ion exchanges. Electrostatic interaction, ion exchange, and bonding to functional groups may occur orderly in the Pb biosorption process and the formation of pyromorphite (Pb5(PO4)3Cl) contributes to Pb biosorption. About 66 % of the adsorbed Pb was accumulated in the A-B AGS microbial cells. Na2EDTA (0.05 M) can recover 60.3 % of the loaded Pb with the highest microbial activity of granules being remained. All the findings will provide the theoretical basis for the large-scale application of A-B AGS to bioremediate Pb(II)-containing wastewater.