Extracellular Polymeric Substances Content Research Articles

Enhancing sludge methanogenesis with changed micro-environment of anaerobic microorganisms by Fenton iron mud

Conductive materials have been demonstrated to enhance sludge methanogenesis, but few researches have concentrated on the interaction among conductive materials, microorganisms and their immediate living environment. In this study, Fenton iron mud with a high abundance of Fe(III) was recycled and applied in anaerobic reactors to promote anaerobic digestion (AD) process. The results show that the primary content of extracellular polymeric substances (EPS) such as polysaccharides and proteins increased significantly, possibly promoting microbial aggregation. Furthermore, with the increment of redox mediators including humic substances in EPS and Fe(III) introduced by Fenton iron mud, the direct interspecies electron transfer (DIET) between methanogens and interacting bacteria could be accelerated, which enhanced the rate of methanogenesis in anaerobic digestion (35.21 ± 4.53% increase compared to the control). The further analysis of the anaerobic microbial community confirmed the fact that Fenton iron mud enriched functional microorganisms, such as the abundance of CO2-reducing (e.g. Chloroflexi) and Fe(III)-reducing bacteria (e.g., Tepidimicrobium), thereby expediting the electron transfer reaction in the AD process via microbial DIET and dissimilatory iron reduction (DIR). This work will make it possible for using the recycled hazardous material – Fenton iron mud to improve the performance of anaerobic granular sludge during methanogenesis.

A new easy method for determination of surface adhesion of phototrophic biofilms.

Terrestrial cyanobacteria grow as phototrophic biofilms and offer a wide spectrum of interesting products. For cultivation of phototrophic biofilms different reactor concepts have been developed in the last years. One of the main influencing factors is the surface material and the adhesion strength of the chosen production strain. In this work a flow chamber was developed, in which, in combination with optical coherence tomography and computational fluid dynamics simulation, an easy analysis of adhesion forces between different biofilms and varied surface materials is possible. Hereby, differences between two cyanobacteria strains and two surface materials were shown. With longer cultivation time of biofilms adhesion increased in all experiments. Additionally, the content of extracellular polymeric substances was analyzed and its role in surface adhesion was evaluated. To test the comparability of obtained results from the flow chamber with other methods, analogous experiments were conducted with a rotational rheometer, which proved to be successful. Thus, with the presented flow chamber an easy to implement method for analysis of biofilm adhesion was developed, which can be used in future research for determination of suitable combinations of microorganisms with cultivation surfaces on lab scale in advance of larger processes.

Characteristics variations of size-fractionated anammox granules and identification of the potential effects on these evolutions

Anaerobic ammonium oxidation (anammox) granulation which contributed to system stabilization and performance improvement has great potential in the field of wastewater nitrogen removal. The researchers fractionated anammox granules into small-size (0.5–0.9 mm), medium-size (1.8–2.2 mm), and large-size (2.8–3.5 mm) categories to examine their properties and mechanisms. Various analyses, including high-throughput sequencing, determination of inorganic elements and extracellular polymeric substances (EPS), and microbial function prediction, were conducted to characterize these granules and understand their impact. The results revealed distinct characteristics among the different-sized granules. Medium-size granules exhibited the highest sphericity, EPS content, and anammox abundance. In contrast, large-size granules had the highest specific surface area, heme c content, specific anammox activity, biodiversity, and abundance of filamentous bacteria. Furthermore, the precipitates within the granules were identified as CaCO3 and MgCO3, with the highest inorganic element content found in the large-size granules. Microbial community and function annotation also varied with granule size. Based on systematic analysis, the researchers concluded that cell growth, chemical precipitation, EPS secretion, and interspecies interaction all played a role in granulation. Small-size granules were primarily formed through cell growth and biofilm formation. As granule size increased, EPS secretion and chemical precipitation became more influential in the granulation process. In the large-size granules, chemical precipitation and interspecies interaction, including synergistic effects with nitrifying, denitrifying, and filamentous bacteria, as well as metabolic cross-feeding, played significant roles in aggregation. This interplay ultimately contributed to higher anammox activity in the large-size granules. By fully understanding the mechanisms involved in granulation, this study provides valuable insights for the acclimation of anammox granules with optimal sizes under different operational conditions.

Enrichment of biofertilizer-type hydrogen-oxidizing bacteria on urea containing Cu(II)

With the utilization of pesticides and fertilizers (e.g. urea), the presence of nitrogen and heavy metals (e.g. copper) can enter and pollute the environment. Biofertilizers can be used to replace chemical fertilizers to increase crop yields and reduce environmental stress. The utilization of hydrogen-oxidizing bacteria (HOB) to be biofertilizers has recently attracted more attention. However, the enrichment of HOB on urea and the effect of copper are undetermined. HOB were successfully enriched using urea in this investigation. The average urea conversion rate (AUCR) was 180.08 mgN/L/d with a hydraulic retention time of 10 h. Microbial community (R1) was dominated by Hydrogenophaga (83.92%), a biofertilizer-type HOB. After addition of 5.47 mg/L Cu2+, the AUCR was decreased by 16%–151.18 mgN/L/d, and the growth of HOB was inhibited by 48%. Meanwhile, inhibition was also reflected by the increase of polysaccharide content (20.27 ± 0.57 to 33.45 ± 2.53 mg/gVSS) and protein content (106.19 ± 19.39 to 125.14 ± 24.73 mg/gVSS) of extracellular polymeric substances in the HOB. The resulting microbial community (R2) was changed to Azospiralium-dominated flora (91.33%). Both enriched microbial communities (R1 and R2) exhibited the abilities of ACC degradation and phosphate solubilization. This study demonstrates that employing urea can directly enrich biofertilizer-type HOB and copper-tolerant HOB can be obtained in a 5.47 mg/L Cu2+ environment. The results provide potential methods to obtain biofertilizer from copper-containing urea wastewater via HOB.

Role of Nano-Fe3O4 for enhancing nitrate removal in microbial electrolytic cells: Characterizations and microbial patterns of cathodic biofilm

Conductive magnetite nanoparticle (Nano-Fe3O4) can facilitate numerous biological reduction reactions as an outstanding electron mediator for electron transfer. The positive role of Nano-Fe3O4 for nitrate removal has gradually gained attention recent years, however, it has not been clarified for the persistence of the promoting effect under different concentrations addition. Performance of nitrogen removal and characteristics of cathodic biofilm were evaluated in this study after Nano-Fe3O4 addition with gradient concentration of 100∼500 mg L−1 in microbial electrolytic cells (MEC). Our study illustrated that the optimal concentration was 200 mg L−1 as the removal rate of nitrate increased by 24.76% and the removal rate of total dissolved nitrogen by 29.72%. At the optimal concentration, Nano-Fe3O4 increased cathodic biofilm DNA concentration by 61.04%, enhanced electron transport system activity, enriched iron redox bacteria, denitrifying bacteria and genes, as well as increased extracellular polymeric substances (EPS) amount, especially the protein content of soluble-EPS. However, promoting effect on nitrate removal was not visible in high concentration (500 mg L−1) addition, its electron transport system activity and EPS content were even declined. XPS results indicated that high concentration of Nano-Fe3O4 may reduce the availability of electrons to cathodic biofilm by competing for electrons, which inhibit nitrate removal.

Effect of micronutrient iron on bioactive compounds isolated from cryptophytes.

Iron is one of the important micronutrients affecting algal growth due to its fundamental role in the physiological processes, including photosynthetic electron transport, respiration, and nitrogen fixation. In this study, the effect of different iron levels on growth and the production of bioactive compounds (phycoerythrin (PE), extracellular polymeric substances (EPS), and phenolic compounds (PCs)) of five cryptophyte strains were investigated. Also, the antioxidant capacity of the bioactive compounds was explored. The results showed species-specific responses to the impact of iron on growth of cryptophytes and accumulation of bioactive compounds. The growth rates of C. pyrenoidifera and Cryptomonas sp. varied significantly at different iron levels, and a reduction in the PE content was observed for several cryptophytes cultured at the highest iron level. However, no significant differences were detected in EPS content at different iron levels. Differences in PC contents of C. pyrenoidifera and Cryptomonas sp. at medium iron level were statistically significant compared with the other two treatments. The results also revealed species-specific differences in antioxidant activity at different iron levels; each studied strain followed its own pattern in response to change in iron level, and each bioactive compound had a different antioxidant activity. Overall, however, PCs demonstrated higher antioxidant activity than PE and EPS. In summary, iron has an impact on growth, bioactive compound accumulation, and antioxidant activity. However, the species-specific responses to changes in iron level should not be ignored when modifying culture conditions for optimal harvest of bioactive compounds.

Rhamnolipid-induced alleviation of bioclogging in Managed Aquifer Recharge (MAR): Interactions with bacteria and porous media

The prevention and treatment of bioclogging is of great significance to the application of Managed Aquifer Recharge (MAR). This study investigated the alleviating effect of biosurfactant rhamnolipid (RL) on bioclogging by laboratory-scale percolation experiments. The results show that the addition of RL greatly reduced bioclogging. Compared with the group without RL, the relative hydraulic conductivity (K′) of the 100 mg/L RL group increased 5 times at the end of the experiment (23 h), while the bacterial cell amount and extracellular polymeric substances (EPS) content on the sand column surface (0–2 cm) decreased by 60.8% and 85.7%, respectively. In addition, the richness and diversity of the microbial communities within the clogging matter decreased after the addition of RL. A variety of bacterial phyla were found, among which Proteobacteria were predominant in all groups. At the genus level, RL reduced the relative abundance of Acinetobacter, Bacillus, Klebsiella, and Pseudomonas. These microbes are known as strong adhesion, large size, and easy to form biofilms, therefore playing a critical role during MAR bioclogging. Moreover, RL changed the surface properties of bacteria and porous media, which results in the increase of electrostatic repulsion and decrease of hydrophobic interaction between them. Therefore, RL mediated the bacteria-porous media interaction to reduce biomass in porous media, thereby alleviating bioclogging. This study implies that RL's addition is an environmentally friendly and effective method to alleviate the bioclogging in MAR.

Revealing the Bacterial Quorum-Sensing Effect on the Biofilm Formation of Diatom Cylindrotheca sp. Using Multimodal Imaging.

Diatoms contribute to carbon fixation in the oceans by photosynthesis and always form biofouling organized by extracellular polymeric substances (EPS) in the marine environment. Bacteria-produced quorum-sensing signal molecules N-acyl homoserine lactones (AHLs) were found to play an important role in the development of Cylindrotheca sp. in previous studies, but the EPS composition change was unclear. This study used the technology of alcian blue staining and scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to directly observe the biofilm formation process. The results showed that AHLs promote the growth rates of diatoms and the EPS secretion of biofilm components. AHLs facilitated the diatom-biofilm formation by a forming process dependent on the length of carbon chains. AHLs increased the biofilm thickness and the fluorescence intensity and then altered the three-dimensional (3D) structures of the diatom-biofilm. In addition, the enhanced EPS content in the diatom-biofilm testified that AHLs aided biofilm formation. This study provides a collection of new experimental evidence of the interaction between bacteria and microalgae in fouling biofilms.

Microplastic biofilm: An important microniche that may accelerate the spread of antibiotic resistance genes via natural transformation

Microplastic (MP) biofilms provide a specific microniche for microbial life and are a potential hotspot for the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs). Nevertheless, the acquisition of ARGs in MP biofilms via natural transformation mediated by extracellular DNA (eDNA) has been rarely explored. This study demonstrated that MP biofilms promoted the natural transformation of extracellular ARGs at the single-cell and multi-species levels, compared to natural substrate (NS) biofilms and bacterioplankton. The transformation frequency on MP biofilms was up to 1000-fold compare to that on NS. The small MPs and aged MPs enhanced the ARG transformation frequencies up to 77.16-fold and 32.05-fold, respectively, compared with the large MPs and pristine MPs. The transformation frequencies on MP biofilms were significantly positively correlated with the bacterial density and extracellular polymeric substance (EPS) content (P < 0.05). Furthermore, MPs significantly increased the expression of the biofilm formation related genes (motA and pgaA) and DNA uptake related genes (pilX and comA) compared to NS and bacterioplankton. The more transformants colonized on MPs contributed to the enhanced transformation frequencies at the community-wide level. Overall, eDNA-mediated transformation in MP biofilms may be an important path of ARG spread, which was promoted by heterogeneous biofilm.

The rhizosphere soil properties and bacteria community of poplar are affected by magnetic field under salt condition

Magnetic fields (MFs) are konwn to affect plant growth, but their effects on soil conditions remain obscure. In this study, poplar seedlings (Populus×euramericana 'Neva') were subjected to a MF under saline conditions, and the rhizosphere soil properties and bacterial communities were then examined. Salt stress detrimentally influenced rhizosphere soil nutrients availability and bacterial community structure. However, MF treatments showed several positive effects, such as increasing the ammonium nitrogen (NH4+-N), amorphous iron oxide (Feo) and available phosphorus (AP) contents in the soil. Moreover,. soil urease and acid phosphatase activity increased by 59% and 63%, respectively. Meanwhile, MF treatments significantly improved the contents of extracellular polymeric substances (EPS) in rhizosphere soils and increased the relative abundance of dominant bacteria phyla (e.g., Actinobacteria, Gemmatimonadetes, and Proteobacteria). Furthermore, a co-occurrence network analysis revealed greater modular clustering in the MF treatments, however, the network stability did not improve. In conclusion, MFs increased rhizobacterial community modularity and promoted the production of bacterial EPS, these effects that then enhanced soil nutirent availability and poplar salt tolerance.

Responses of halophilic microbial communities to changes in salt composition: Comparison between autotrophic nitrification and heterotrophic ammonia assimilation biosystems

The concentration and proportion of chlorine (Cl−) and sulfate ions (SO42−) in actual high salinity wastewater exhibit significant fluctuations due to their diverse sources. This study compared the response of halophilic autotrophic nitrification (AN) and heterotrophic ammonia assimilation (HAA) sludges to changes in salt composition. The results demonstrated that both the AN and HAA systems maintained high ammonia removal efficiency even when exposed to mixed salt ions or pure sulfate conditions. Increasing the concentration of SO42− resulted in an increase in extracellular polymeric substances content, sludge settleability, sludge hydrophobicity, and the relative abundance of Nitrosomonas in the AN system (from 2.3% to 10.4%). The dominant heterotrophic bacteria in the HAA system underwent turnover in response to changes in salt composition conditions. The robustness and the cooperation between microorganisms of the HAA system surpassed those of the AN system. This study provides scientific foundation for treating multi-ion high salinity wastewater.

Change in Granulation Potential and Microbial Enrichment Characteristics of Sludge Induced by Microplastics

Microplastics (MPs), as a new type of pollutant, are widely detected in sewage treatment plants. Currently, research on MPs in traditional sewage treatment systems has mainly been focused on the pollution level and distribution characteristics, with a lack of studying the impact of MPs on the sludge granulation. In order to explore the effect of MPs on the granulation process, a microplastic exposure test was conducted by adding polyethylene terephthalate microplastics (PET-MPs), which are widespread in the environment. The operating performance of the system, extracellular polymeric substance (EPS) composition, and flora enrichment were analyzed on the sludge granulation. The results showed that the exposure of PET-MPs significantly accelerated the sludge granulation process, whereas the increase in EPS content dominated by PN enhanced the sludge surface hydrophobicity; the granulation rate and EPS secretion were proportional to the exposed particle size. Microplastics and EPS secretions synergistically promoted the formation of granular sludge. However, continuous microplastic exposure led to deterioration of the system decontamination performance and inhibited the degradation process of pollutants, with the most negative effect of nitrite nitrogen accumulation under 250 μm PET-MPs exposure, as high as (5.08±0.24) mg·L-1. The high-throughput sequencing revealed that the microbial community diversity fell in the experimental group. The dominant bacteria at the phylum level were Proteobacteria and Bacteroidota on the sludge granulation. Rhodocyclaceae, Sphingomonadaceae, Flavobacteriaceae, and Rhodanobacteraceae promoted flocculation by increasing EPS secretion. The decrease in Comamonadaceae and Chitinophagaceae weakened the ammonia and nitrite oxidation capacity of the system, whereas the decrease in Rhodobacteraceae, Hyphomonadaceae, and Xanthomonadaceae inhibited the removal of nitrate nitrogen.

Insights into the response of Anammox sludge to the individual and combined stress of neodymium and salinity and the role of hydroxylamine regulation

Neodymium (Nd(III)) and sulfate (Na2SO4) are commonly present in neodymium mining wastewater. However, the effects of their coexistence on microorganisms responsible for removing nitrogen from wastewater have been poorly reported. In this study, the combined inhibition of Nd(III) and Na2SO4 on anaerobic ammonium oxidation (Anammox) and its recovery were investigated in terms of performance, kinetics, Nd(III) distribution characteristics and sludge morphology. The results shown that ≥ 150 mg·L−1 Nd(III) or ≥ 3 g·L−1 Na2SO4 decreased specific Anammox activity (SAA), and intracellular Nd(III) was identified as the primary factor for individual Nd(III) inhibition. Furthermore, the combined inhibition of Nd(III) and Na2SO4 on Anammox was stronger than individual inhibition, but low doses of hydroxylamine (NH2OH) could restore the Anammox. Notably, the combined effect of low doses of Na2SO4 (1 g·L−1) and Nd III (10 mg·L−1) on the long-term performance of Anammox reactor was not significant. However, when the dose of Nd(III) was kept constant and the dose of Na2SO4 was increased to 7 g·L−1, the total nitrogen removal efficiency (TNRE) decreased by 56.88%. Following the elimination of inhibitors and the addition of 5 mg·L−1 NH2OH, the TNRE of the Anammox reactor was increased from 20.83% to 29.78%. The Hill, modified Michaelis-Menten, and modified Boltzmann models were able to properly describe the inhibition effects of Nd(III) and Na2SO4 and recovery process of Anammox. In addition, scanning electron microscopy (SEM) and energy-dispersive X-ray (EDS) analysis revealed that the content of extracellular polymeric substances (EPS) and neodymium increased with increasing Na2SO4 concentration.

Comparative analysis of membrane fouling mechanisms induced by operation modes of membrane bioreactors with aerobic granular sludge

This experimental work investigated fouling characteristics induced by two different configurations of membrane bioreactor (MBR), which are submerged MBR and sidestream MBR with aerobic granular sludge. Submerged membrane bioreactor with granular sludge (Sub-MGSBR) ran the longest operation time 61 days with a steady overall TMP increase rate; Sidestream membrane bioreactor with granular sludge (SS-MGSBR) performed only 39 days, which exhibited Sub-MGSBR had more efficiently retarding membrane fouling. In both membrane bioreactors with flocculent sludge (MFSBRs) as a control, membrane foulants were compact, and cake resistance was the dominant fouling factor. In MGSBRs, however, pore blocking resistance turned out the key fouling factor. Especially in Sub-MGSBR, it went beyond 75%, and there was the most conglomeration of microorganisms of foulants with the highest porosity. Extracellular polymeric substances (EPS) content of foulants proved membrane fouling was hardly just for granules accumulation into cake but microorganisms’ growth in MGSBRs.

The presence of copper ions alters tetracycline removal pathway in aerobic granular sludge: Performance and mechanism

This study investigated the removal characteristics of tetracycline (TC) in the presence of copper ions (Cu2+) in aerobic granular sludge by analyzing the TC removal pathway, composition and functional group changes of extracellular polymeric substances (EPS), and microbial community structure. The TC removal pathway changed from cell biosorption to EPS biosorption, and the microbial degradation rate of TC was reduced by 21.37% in the presence of Cu2+. Cu2+ and TC induced enrichment of denitrifying bacteria and EPS-producing bacteria by regulating the expression of signaling molecules and amino acid synthesis genes to increase the content of EPS and –NH2 groups in EPS. Although Cu2+ reduced the content of acidic hydroxyl functional groups (AHFG) in EPS, an increase in TC concentration stimulated the secretion of more AHFG and –NH2 groups in EPS. The long-term presence of TC presence of the relative abundances of Thauera, Flavobacterium and Rhodobacter and improved the removal efficiency.

Starvation combined with constant anode potential triggers intracellular electron storage in electro-active biofilms

The accumulation of electrons in the form of Extracellular Polymeric Substances (EPS) and poly-hydroxyalkanoates (PHA) has been studied in anaerobic processes by adjusting the access of microorganisms to the electron donor and final electron acceptor. In Bio-electrochemical systems (BESs), intermittent anode potential regimes have also recently been used to study electron storage in anodic electro-active biofilms (EABfs), but the effect of electron donor feeding mode on electron storage has not been explored. Therefore, in this study, the accumulation of electrons in the form of EPS and PHA was studied as a function of the operating conditions. EABfs were grown under both constant and intermittent anode potential regimes and fed with acetate (electron donor) continuously or in batch. Confocal Laser Scanning Microscopy (CLSM) and Fourier-Transform Infrared Spectroscopy (FTIR) were used to assess electron storage. The range of Coulombic efficiencies, from 25 to 82%, and the biomass yields, between 10 and 20%, indicate that storage could have been an alternative electron consuming process. From image processing, a 0.92 pixel ratio of poly-hydroxybutyrate (PHB) and amount of cells was found in the batch fed EABf grown under a constant anode potential. This storage was linked to the presence of living Geobacter and shows that energy gain and carbon source starvation were the triggers for intracellular electron storage. The highest EPS content (extracellular storage) was observed in the continuously fed EABf under an intermittent anode potential, showing that constant access to electron donor and intermittent access to the electron acceptor leads to the formation of EPS from the excess energy gained. Tailoring operating conditions can thus steer the microbial community and result in a trained EABf to perform a desired biological conversion, which can be beneficial for a more efficient and optimized BES.

Understanding the cometabolic degradation of sulfadiazine by an enriched ammonia oxidizing bacteria culture from both extracellular and intracellular perspectives

Antibiotics are widely used drugs in the world and pose serious threats to ecosystems and human health. Although it has been reported that ammonia oxidizing bacteria (AOB) can cometabolize antibiotics, little has been reported on how AOB would respond to the exposure of antibiotics on extracellular and enzymatic levels, as well as the impact of antibiotics on the bioactivity of AOB. Therefore, in this study, a typical antibiotic, sulfadiazine (SDZ), was selected, and a series short-term batch tests using enriched AOB sludge were conducted to investigate the intracellular and extracellular responses of AOB along the cometabolic degradation process of SDZ. The results showed the cometabolic degradation of AOB made the main contribution to SDZ removal. When the enriched AOB sludge was exposed to SDZ, ammonium oxidation rate, ammonia monooxygenase activity, adenosine triphosphate concentration and dehydrogenases activity were negatively affected. The amoA gene abundance increased 1.5 folds within 24 h, which may enhance the uptake and utilization of substrates and maintain stable metabolic activity. In the tests with and without ammonium, the concentration of total EPS increased from 264.9 to 231.1 mg/gVSS to 607.7 and 538.2 mg/gVSS, respectively, under the exposure to SDZ, which was mainly contributed by the increase of proteins in tightly bound extracellular polymeric substances (EPS) and polysacharides in tightly bound EPS and soluble microbial products. The proportion of tryptophan-like protein and humic acid-like organics in EPS also increased. Moreover, SDZ stress stimulated the secretion of three quorum sensing signal molecules, C4-HSL (from 140.3 to 164.9 ng/L), 3OC6-HSL (from 17.8 to 42.4 ng/L) and C8-HSL (from 35.8 to 95.9 ng/L) in the enriched AOB sludge. Among them, C8-HSL may be a key signal molecule that promoted the secretion of EPS. The findings of this study could shed more light on the cometabolic degradation of antibiotics by AOB.

Response of nitrite accumulation to elevated C/NO– 3-N ratio during partial denitrification process: Insights of extracellular polymeric substance, microbial community and metabolic function

This study investigated the response of nitrite accumulation to elevated COD/NO3–-N ratio (C/N) during partial denitrification (PD). Results indicated nitrite was gradually accumulated and remained stable (C/N = 1.5 ∼ 3.0), while that rapidly declined after reaching the peak (C/N = 4.0 ∼ 5.0). The polysaccharide (PS) and protein (PN) content of tightly-bound extracellular polymeric substances (TB-EPS) reached the maximum at C/N of 2.5 ∼ 3.0, which might be stimulated by high level of nitrite. Illumina MiSeq sequencing showed Thauera and OLB8 were dominated denitrifying genera at C/N of 1.5 ∼ 3.0, while Thauera was further enriched with fading OLB8 at C/N of 4.0 ∼ 5.0. Meanwhile, the highly-enriched Thauera might enhance the activity of nitrite reductase (nirK) promoting further nitrite reduction. Redundancy analysis (RDA) showed positive correlations between nitrite production and PN content of TB-EPS, denitrifying bacteria (Thauera and OLB8) and nitrate reductases (narG/H/I) in low C/N. Finally, their synergistic effects for driving nitrite accumulation were comprehensively elucidated.

Anaerobic fermentation of aerobic granular sludge: Insight into the effect of granule size and sludge structure on hydrolysis and acidification

Aerobic granular sludge (AGS) has different physicochemical properties and microbial communities compared to conventional activated sludge (CAS), which may result in different behaviors during anaerobic fermentation and require further investigation. This study investigated the effect of granule size and sludge structure on the hydrolysis and acidification of AGS. Experimental results show that AGS exhibited significantly higher soluble chemical oxygen demand (SCOD) dissolution and total volatile fatty acids (TVFA) production (330.6–430.3 mg/gVSS and 231.0–312.5 mgCOD/gVSS) compared to conventional activated sludge (CAS) (167.0 mg/gVSS and 133.3 mgCOD/gVSS). This is because AGS (90.6–96.9 mg/gVSS) had higher extracellular polymeric substances (EPS) content than CAS (81.2 mg/gVSS). EPS can not only serve as substrates but also release the trapped hydrolases. Moreover, the relative abundances of hydrolytic/acidogenic bacteria and genes were higher in AGS (0.46%–3.60% and 3.01 × 10−3%-4.04 × 10−3%) than in CAS (0.30% and 1.23 × 10−3%). The optimal granule size for AGS fermentation was found to be 500–1600 μm. The crushing of granule structure promoted the dissolution of small amounts of EPS and the release of some trapped hydrolases, thereby potentially enhancing the enzyme-substrate contacts and bacteria-substrate interactions. Therefore, the highest SCOD dissolution (510.6 mg/gVSS) and TVFA production (352.1 mgCOD/gVSS) from crushed 500–1600 μm AGS were observed. Overall, the findings of this study provide valuable insights into the recovery of organic carbon from AGS via anaerobic fermentation.

Long-term operation and biofouling of graphene oxide membrane in practical water treatment: Insights from performance and biofilm characteristics

The widespread research of graphene oxide (GO) membranes has revealed the need to investigate their long-term performance and associated biofouling behavior in practical water treatment conditions. In this study, we have evaluated the long-term operation of GO membranes from the perspective of filtration performance and biofilm characteristics, based on two representative GO membranes (V-GO and P-GO, fabricated by vacuum and pressure filtration, respectively) with different surface properties and a natural surface water, using a gravity driven membrane system. The results showed that the GO membranes were capable of achieving sustainable water purification over 45 days of operation, associated with a complete rejection for biopolymers and desirable removal for fluorescent organic matters. Furthermore, the V-GO membrane with a rougher surface, reduced hydrophilicity and higher sheet-edge exposure than that of the P-GO membrane formed a highly heterogeneous and porous biofilm with high permeability, despite its greater biofilm thickness and extracellular polymeric substances (EPS) contents. In contrast, the P-GO membrane biofilm exhibited a thin but dense structure, and therefore an increased hydraulic resistance (∼1.5 times greater than V-GO membrane). The microbial community analysis revealed that bacteria related to biofilm formation was richer in V-GO biofilm. While bacteria associated with the degradation of organic matters and the aggregation of microorganisms accounted for a greater proportion in P-GO biofilm. These factors were responsible for forming a thick biofilm for V-GO, and a thin but high resistance biofilm for P-GO membrane. Our findings highlight the sustainable water purification performance of the GO membranes, and the P-GO membrane can alleviate biofilm formation but not necessarily reduce biofouling during the long-term filtration, while this was opposite in case of the V-GO membrane.