Sludge Microorganisms Research Articles

Microbial transformation of lignite into methane: Insights from anaerobic-activated sludge systems

Lignite pretreated by H2O2 was degraded by microorganisms in anaerobic activated sludge to produce methane, and the microorganisms were analyzed by high-throughput techniques. Samples fermented for 60 days were analyzed by high performance liquid chromatography (HPLC) and gas chromatography–mass spectrometry (GC–MS). Methane production was found to reach a maximum rate of 2.25 mL/g·d−1 at pH 7 and 35 °C, which was synchronized with the efficient consumption of organic acids, with a peak organic acid production of 202.13 COD/mg·L−1. A total of 37 compounds including alkanes, ketones, esters and polycyclic aromatic hydrocarbons were detected in the fermentation broth, and the species compositions of anaerobic sludge microbiota were fungal, bacterial and archaeal are Ascomycota, Bacteroidetes and Methanosaeta respectively in the fermentation system. Based on the analysis of above results, it is initially speculated that the biochemical pathway for lignite degradation and methane production by anaerobic sludge microorganisms is the acetate pathway.

Nanostructured materials as hazardous wastewater micropullutants: Sources, behavior, and impact on functional bacterial community of activated sludge

Unique properties of nanoscale materials make them attractive for industrial, medical, agricultural, and environmental applications. Nevertheless, the release of nanoparticles into the environment is a major concern due to the lack of knowledge about their behavior in the environment and potential widespread environmental impacts. On the one hand, nanomaterials are perceived as pollutants that may affect activated sludge microorganisms and, consequently, the efficiency of wastewater treatment processes. On the other hand, some nanomaterials can be intentionally added to activated sludge systems to improve their performance in terms of, e.g., sludge settling and removing heavy metals or organic pollutants. As a result, nanoparticles are frequently accumulated in wastewater, which is considered to be a major source of nanoparticle release to the surrounding environment. Processes that involve the action of activated sludge are used worldwide in wastewater treatment plants due to their excellent capacity of removing nutrients, degrading toxins, and retaining biomass. High concentrations of nanoparticles entering activated sludge systems can affect their growth and metabolism. The research studies, which are reviewed in the present article, show that nanoparticles significantly reduce the relative abundance of the activated sludge microbial community associated with nitrification, denitrification, and phosphorus removal. The knowledge about the structure of the activated sludge microbial community with an assessment of nanomaterial toxicity can contribute to optimizing the sludge population and improving the performance of wastewater treatment plants.

The predicted secreted proteome of activated sludge microorganisms indicates distinct nutrient niches.

In wastewater treatment plants (WWTPs), complex microbial communities process diverse chemical compounds from sewage. Secreted proteins are critical because many are the first to interact with or degrade external (macro)molecules. To better understand microbial functions in WWTPs, we predicted secreted proteomes of WWTP microbiota from more than 1,000 high-quality metagenome-assembled genomes (MAGs) from 23 Danish WWTPs with biological nutrient removal. Focus was placed on examining secreted catabolic exoenzymes that target major classes of macromolecules. We demonstrate that Bacteroidota has a high potential to digest complex polysaccharides, but also proteins and nucleic acids. Poorly understood activated sludge members of Acidobacteriota and Gemmatimonadota also have high capacities for extracellular polysaccharide digestion. Secreted nucleases are encoded by 61% of MAGs indicating an importance for extracellular DNA and/or RNA digestion in WWTPs. Secreted lipases were the least common macromolecule-targeting enzymes predicted, encoded mainly by Gammaproteobacteria and Myxococcota. In contrast, diverse taxa encode extracellular peptidases, indicating that proteins are widely used nutrients. Diverse secreted multi-heme cytochromes suggest capabilities for extracellular electron transfer by various taxa, including some Bacteroidota that encode undescribed cytochromes with >100 heme-binding motifs. Myxococcota have exceptionally large secreted protein complements, probably related to predatory lifestyles and/or complex cell cycles. Many Gammaproteobacteria MAGs (mostly former Betaproteobacteria) encode few or no secreted hydrolases, but many periplasmic substrate-binding proteins and ABC- and TRAP-transporters, suggesting they are mostly sustained by small molecules. Together, this study provides a comprehensive overview of how WWTPs microorganisms interact with the environment, providing new insights into their functioning and niche partitioning.IMPORTANCEWastewater treatment plants (WWTPs) are critical biotechnological systems that clean wastewater, allowing the water to reenter the environment and limit eutrophication and pollution. They are also increasingly important for the recovery of resources. They function primarily by the activity of microorganisms, which act as a "living sponge," taking up and transforming nutrients, organic material, and pollutants. Despite much research, many microorganisms in WWTPs are uncultivated and poorly characterized, limiting our understanding of their functioning. Here, we analyzed a large collection of high-quality metagenome-assembled genomes from WWTPs for encoded secreted enzymes and proteins, with special emphasis on those used to degrade organic material. This analysis showed highly distinct secreted proteome profiles among different major phylogenetic groups of microorganisms, thereby providing new insights into how different groups function and co-exist in activated sludge. This knowledge will contribute to a better understanding of how to efficiently manage and exploit WWTP microbiomes.

Carbon chain elongation characterizations of electrode-biofilm microbes in electro-fermentation

The higher efficiency of electro-fermentation in synthesizing medium-chain fatty acids (MCFAs) compared to traditional fermentation has been acknowledged. However, the functional mechanisms of electrode-biofilm enhancing MCFAs synthesis remain research gaps. To address this, this study proposed a continuous flow electrode-biofilm reactor for chain elongation (CE). After 225 days of operation, stable electrode-biofilms formed and notably improved caproate yield by more than 38 %. The electrode-biofilm was enriched with more CE microorganisms and electroactive bacteria compared to the suspended sludge microorganisms, including Caproicibacterium, Oscillibacter and Pseudoramibacter. Besides, the upregulated CE pathways were evaluated by metagenomic analysis, and the results indicated that the pathways such as acetyl-CoA and malonyl-[acp] formation, reverse beta-oxidation, and fatty acid biosynthesis pathway were all markedly enhanced in cathodic biofilm, more than anodic biofilm and suspended microorganisms. Moreover, microbial community regulated processes like bacterial chemotaxis, flagellar assembly and quorum sensing, crucial for electrode-biofilm formation. Electron transfer, energy metabolism, and microbial interactions were found to be prominently upregulated in the cathodic biofilm, surpassing levels observed in anodic biofilm and suspended sludge microorganisms, which further enhanced CE efficiency. In addition, the statistical analyses further highlighted key microbial functions and interactions within the cathodic biofilm. Oscillospiraceae_bacterium was identified to be the most active microbe, alongside pivotal roles played by Caproiciproducens_sp._NJN-50, Clostridiales_bacterium, Prevotella_sp. and Pseudoclavibacter_caeni. Eventually, the proposed microbial collaboration mechanisms of cathodic biofilm were ascertained. Overall, this study uncovered the biological effects of the electrode-biofilm on MCFAs electrosynthesis, thereby advancing biochemicals production and filling the knowledge gaps in CE electroactive biofilm reactors.

A yak gut probiotic Lactobacillus paracasei T1-9 displays superior degradation of poly(butylene adipate-co-terephthalate) bioplastic

Poly(butylene adipate-co-terephthalate) (PBAT) has gained significant attention for its exceptional processing properties and biodegradability. However, PBAT displays low biodegradability in natural environment. Many studies found degradable microorganisms in wastewater sludge, soil, compost, etc., but most are harmful to humans. This work aimed to explore the potential degradation of PBAT by probiotics. We screened 47 kinds of safety microbes for PBAT degradation, five probiotics showed positive degradation effects on PBAT. Among these, Lactobacillus paracasei T1–9 exhibited superior ability to degrade PBAT, achieving the highest percentage of weight loss at 1.77 ± 0.08 %, along with highly efficient growth in liquid culture. The biodegradability of PBAT was evaluated by using a multifaceted approach encompassing techniques including SEM, FTIR, XPS, and LC-MS. To improve the degradation efficiency, various factors (pH, the addition of gelatin and carbon source) were investigated. The additional gelatin improved the degradation of PBAT at a 3.43 ± 0.1 % weight loss. As the carbon source in medium, 1, 4-butanediol contributed the highest biodegradation effect compared to the other two monomers of PBAT. Interestingly, the supernatants of T1–9 incubated with PBAT displayed the highest lipase activity with 3.99 ± 0.03 U/mL. In conclusion, the probiotic T1–9 processed excellent capabilities in degrading PBAT, with the primary enzyme hypothesized to belong to the lipase group.

Integration of mixotrophic denitrification and anammox in Thiothrix-hydroxyapatite coupled particles treating municipal wastewater against low temperature and organic load shocks

Sulfur-driven autotrophic denitrification coupled with anammox (SPDA) is a promising process for autotrophic nitrogen removal, but autotrophic microorganisms are challenging to retain in wastewater treatment units. In this study, a single-stage mixotrophic denitrification coupled with anammox (MDA) process was established in an up-flow anaerobic sludge bed reactor with Thiothrix-hydroxyapatite coupled particles. The advancement of the sludge granulation process effectively strengthened the resistance of the MDA system to environmental stresses, and the SPDA-dominated nitrogen removal process achieved a total nitrogen removal efficiency of 88.6 %, a nitrogen removal rate of 0.432 KgN/m3/d at temperatures as low as 12.8 °C and an organic loading rate of 0.63 KgCOD/m3/d. Interestingly, the ecological niche integration of sulfur oxidizing bacteria (SOB), anammox bacteria and heterotrophic denitrifying bacteria in the granular sludge was observed during long-term operation. The FISH-CLSM results revealed the spatial distribution of functional microorganisms in the granular sludge and the potential pollutant transformation mechanism. The aggressive growth of SOB dominated by Thiothrix constructed a biological wall at the periphery of the granules, which ensured their priority for nitrate acquisition, supplied nitrite to the internal anammox bacteria, and acted as an important barrier against the shock of organic loads. Overall, this study presents a novel granulation approach for the biological treatment of sulfur-containing wastewater, highlighting the efficacy of functional integration within Thiothrix-hydroxyapatite coupled particles as an effective strategy to resist environmental stress.

A novel three-dimensional biofilm-electrode/anoxic-oxic process for nitrogen and phosphorus removal: Performance, mechanism and microbial community

Traditional anoxic-oxic (AO) process cannot meet increasingly high pollutants emission standards in China. In order to improve removal efficiencies of the pollutants and the electron transfer rate, graphite-graphite electrodes and iron carbon fillers were added into AO reactor to form three-dimensional biofilm electrode/anoxic aerobic reactor (3DBER/AO). Removal efficiencies of the pollutants, sludge properties and microbial community structure were investigated under different current intensities. The results showed that microbial activity and removal efficiencies of the pollutants were the maximum, and mass percentage of Fe reached the highest value of 5.11 % under the current of 40 mA. Excessive or insufficient current intensity could lead to a decrease in sludge activity and removal efficiencies of the pollutants. The removal of nitrogen mainly depended on the nitrification and denitrification of microorganisms, and dissolved phosphorus was removed through chemical precipitation. Current intensity affected microbial communities and biosynthesis. AKYH767 and Saccharmonadales were dominant microorganisms in the cathode and anode sludge, respectively, under the current of 40 mA. However, Azospira and Dechlorobacter were dominant microorganisms in the cathode sludge, and Hydrogenophaga was dominant microorganisms in the anode sludge under the current of 60 mA. These microorganisms were closely related to the removal of organic matter, nitrogen and phosphorus. Therefore, microbial community structure and removal efficiencies of the pollutants could be regulated by controlling the current intensity in the 3DBER/AO process.

Study on the ability of activated sludge bacteria to form biofilms <i>in vitro</i>

The study aims to comparatively characterize in vitro biofilm formation in bacterial cultures isolated from activated sludge, as well as archival cultures capable of xenobiotics biodegradation: Alcaligenes faecalis 2, Acinetobacter guillouiae 11h, Rhodococcus erythropolis ILBIO, and Achromobacter pulmonis PNOS. An analysis of the 16S rRNA nucleotide sequence identified strains isolated from activated sludge: Paenibacillus odorifer, Bacillus subtilis, Micrococcus yunnanensis, and Bacillus proteolyticus. The formation of biofilms by microorganisms was studied on LB medium and synthetic culture medium (with sodium acetate as a carbon source). With cell growth on LB medium, an increase in biofilm biomass was observed in Paenibacillus odorifer, Bacillus subtilis, Alcaligenes faecalis 2, and Achromobacter pulmonis PNOS. The cultivation stage duration (72 and 144 h), as well as the additional dosing of substrates, had an effect on the biofilm formation process: by 144 h of cultivation, the biomass values amounted to 0.6–1.3 optical units. An average 63–77% increase in biofilm biomass was noted for Bacillus subtilis and Paenibacillus odorifer cells as compared to the 72-hour process. At the final stage of cultivation (144 h), the values of exopolysaccharides in the matrix amounted to over 0.02 optical units for Bacillus subtilis and Paenibacillus odorifer. The metabolic activity of activated sludge bacteria forming the biofilm reached 628–3609 Fl./OD540. Thus, activated sludge microorganisms forming the biofilm were shown to retain viability and metabolic activity during growth under in vitro conditions.

From "resistance genes expression" to "horizontal migration" as well as over secretion of Extracellular Polymeric Substances: Sludge microorganism’s response to the increasing of long-term disinfectant stress

Glutaraldehyde-Didecyldimethylammonium bromides (GDs) has been frequently and widely employed in livestock and poultry breeding farms to avoid epidemics such as African swine fever, but its long-term effect on the active sludge microorganisms of the receiving wastewater treatment plant was keep unclear. Four simulation systems were built here to explore the performance of aerobic activated sludge with the long-term exposure of GDs and its mechanism by analyzing water qualities, resistance genes, extracellular polymeric substances and microbial community structure. The results showed that the removal rates of CODCr and ammonia nitrogen decreased with the exposure concentration of GDs increasing. It is worth noting that long-term exposure to GDs can induce the horizontal transfer and coordinated expression of a large number of resistance genes, such as qacE, sul1, tetx, and int1, in drug-resistant microorganisms. Additionally, it promotes the secretion of more extracellular proteins, including arginine, forming a "barrier-like" protection. Therefore, long-term exposure to disinfectants can alter the treatment capacity of activated sludge receiving systems, and the abundance of resistance genes generated through horizontal transfer and coordinated expression by drug-resistant microorganisms does pose a significant threat to ecosystems and health. It is recommended to develop effective pretreatment methods to eliminate disinfectants.

Water reuse from wastewater: Experiences and challenges of implementing a full-scale water reuse system in a food industry plant

This paper presents experiences and challenges related to the implementation and successful 3-year operation of a full-scale station for water reuse from wastewater in one of the largest poultry slaughterhouses in Europe. Solved problems for implementing deammonification, as well as automatic coagulant dosing for wastewater pre-treatment, improvement of activated sludge separation, and digestate management, are presented and discussed. Two major limitations of stable water reclamation were a limit of 30 mg N/L in the effluent to the river and the passing of cationic flocculants and natural polymers formed by activated sludge microorganisms to the ultrafiltration units. The limit of 30 mg N/L was achieved by implementing i) a unique deammonification system to treat leachate from the anaerobic digester, ii) automatic coagulant dosing to mitigate high nitrogen peaks and balance organic compounds between biogas production and efficient denitrification. Flocculants and polymers passing to the ultrafiltration units and causing a sharp drop in water recovery were eliminated by stabilizing the zeta potential of the sludge, adding an antiscalant before the ultrafiltration, and automatic control of wastewater coagulation at primary treatment, responding to COD peaks. With improvements described in the article, the water reuse system provides a stable supply of 4000 m3/d of high-quality water, resulting in over 4 million m3 of water reclaimed since 2020.

The water-soluble fraction of extracellular polymeric substances from a resource recovery demonstration plant: characterization and potential application as an adhesive.

Currently, there is a growing interest in transforming wastewater treatment plants (WWTPs) into resource recovery plants. Microorganisms in aerobic granular sludge produce extracellular polymeric substances (EPS), which are considered sustainable resources to be extracted and can be used in diverse applications. Exploring applications in other high-value materials, such as adhesives, will not only enhance the valorization potential of the EPS but also promote resource recovery. This study aimed to characterize a water-soluble fraction extracted from the EPS collected at the demonstration plant in the Netherlands based on its chemical composition (amino acids, sugar, and fatty acids) and propose a proof-of-concept for its use as an adhesive. This fraction comprises a mixture of biomolecules, such as proteins (26.6 ± 0.3%), sugars (21.8 ± 0.2%), and fatty acids (0.9%). The water-soluble fraction exhibited shear strength reaching 36-51 kPa across a pH range of 2-10 without additional chemical treatment, suggesting a potential application as an adhesive. The findings from this study provide insights into the concept of resource recovery and the valorization of excess sludge at WWTPs.

Potential pathogenic microorganisms in rural wastewater treatment process: Succession characteristics, concentration variation, source exploration, and risk assessment

Pathogenic microorganisms can cause infection, sepsis, and other diseases in humans. Although municipal wastewater plants are important sources and sinks for potential pathogenic microorganisms, data on rural wastewater treatment processes are limited. The proximity of rural wastewater facilities to human settlements and the trend toward wastewater resourcing could pose risks to humans. Here, a typical village in southern China was selected to analyze potential pathogenic microorganisms in wastewater, sewage sludge, and aerosols during the collection, treatment, and discharge of domestic wastewater. The succession characteristics and concentration variations of potential pathogenic microorganisms throughout the wastewater treatment process were identified using high-throughput sequencing and culture methods. Bacteria-associated health risks in facility aerosols were estimated based on average daily dose rates from inhalation and dermal exposure. Lower amounts of pathogenic bacteria and pathogenic fungi were detected in the effluent of the 1-ton treatment scale and the 10-ton treatment scale facilities, compared to those in the influent. Pathogen effluent concentrations were significantly lower than influent concentrations after treatment in rural wastewater facilities. 16 and 29 potential pathogenic bacteria and fungi were detected in aerosols from wastewater treatment facilities, respectively. Furthermore, the potential pathogen concentrations were higher than those in the background air. Aerobic units are the main source of pathogen emissions from aerosols. There were 42 potential pathogenic bacteria and 34 potential pathogenic fungi in the sewage sludge. Biochemical units were the main source of potential pathogens in sewage sludge, and more potential airborne pathogens originated from wastewater. In rural wastewater resourcing processes with greater pollutant exposure, the effluent of rural wastewater treatment facilities (WWTFs), downstream rivers, and facility aerosols, could be important potential sources of microbial risk. Inhalation is the main pathway of human exposure to airborne bacteria. Therefore, more attention should be focused on microbiological risk in rural wastewater treatment processes.

Effect of UV pretreatment on the source control of floR during subsequent biotreatment of florfenicol wastewater.

UV photolysis has been recommended as an alternative pretreatment method for the elimination of antibacterial activity of antibiotics against the indicator strain, but the pretreated antibiotic intermediates might not lose their potential to induce antibiotic resistance genes (ARGs) proliferation during subsequent biotreatment processes. The presence of florfenicol (FLO) in wastewater seriously inhibits the metabolic performance of anaerobic sludge microorganisms, especially the positive correlation between UV irradiation doses and ATP content, while it did not significantly affect the organics utilization ability and protein biosynthetic process of aerobic microorganisms. After sufficient UV pretreatment, the relative abundances of floR from genomic or plasmid DNA in subsequent aerobic and anaerobic biotreatment processes both decreased by two orders of magnitude, maintained at the level of the groups without FLO selective pressure. Meanwhile, the abundances of floR under anaerobic condition were always lower than that under aerobic condition, suggesting that anaerobic biotreatment systems might be more suitable for the effective control of target ARGs. The higher abundance of floR in plasmid DNA than in genome also indicated that the potential transmission risk of mobile ARGs should not be ignored. In addition, the relative abundance of intI1 was positively correlated with floR in its corresponding genomic or plasmid DNA (p < 0.05), which also increased the potential horizontal transfer risk of target ARGs. This study provides new insights into the effect of preferential UV photolysis as a pretreatment method for the enhancement of metabolic performance and source control of target ARGs in subsequent biotreatment processes. KEY POINTS: • Sufficient UV photolytic pretreatment efficiently controlled the abundance of floR • A synchronous decrease in abundance of intI1 reduced the risk of horizontal transfer • An appreciable abundance of floR in plasmid DNA was a potential source of total ARGs.

Constructions of quorum sensing signaling network for activated sludge microbial community.

In wastewater treatment systems, the interactions among various microbes based on chemical signals, namely quorum sensing (QS), play critical roles in influencing microbial structure and function. However, it is challenging to understand the QS-controlled behaviors and the underlying mechanisms in complex microbial communities. In this study, we constructed a QS signaling network, providing insights into the intra- and interspecies interactions of activated sludge microbial communities based on diverse QS signal molecules. Our research underscores the role of diffusible signal factors in both intra- and interspecies communication among activated sludge microorganisms, and signal molecules commonly considered to mediate intraspecies communication may also participate in interspecies communication. QS signaling molecules play an important role as communal resources among the entire microbial group. The communication network within the microbial community is highly redundant, significantly contributing to the stability of natural microbial systems. This work contributes to the establishment of QS signaling network for activated sludge microbial communities, which may complement metabolic exchanges in explaining activated sludge microbial community structure and may help with a variety of future applications, such as making the dynamics and resilience of highly complex ecosystems more predictable.

The influence of ultrasonic exposure on the biocenosis of activated sludge from treatment facilities of domestic and industrial wastewater

Today, in the context of industrial development, an integral part is the formation of wastewater with the need for its subsequent treatment. The biological method of wastewater treatment is the most effective, safe and reliable. This method is based on the work of activated sludge, which is a biocenosis of microorganisms that absorb pollutants from wastewater. Different quantitative and qualitative composition of wastewater and physical impacts can affect the work of the biological agent - activated sludge, therefore research in this area is relevant. To increase the efficiency of biological treatment, a number of methods are used, one of which is ultrasonic exposure of sludge to increase its biochemical activity. However, this approach requires studying its effect on activated sludge microorganisms. In this research work, the effect of ultrasonic exposure on flakes and biocenoses of activated sludge from treatment facilities of various wastewaters was studied and the change in the content of heavy metals after acoustic processing of sludge was determined.

Pollution Monitoring: A Solution to Improve the Quality of Wastewater Treatment from Household Emissions

In the context of the growing environmental pressure on water resources caused by anthropogenic factors, the task of increasing the efficiency of wastewater treatment systems becomes urgent. The purpose of this work is to study and determine the optimal wavelength of the photoemitter for measuring the concentration of activated sludge by the optical method in biological wastewater treatment. The process of interaction of the emitted light with the measured medium is theoretically analyzed, the optical properties of the ultraviolet, visible, and near-infrared regions of the spectrum are considered. Based on these analyzes, an experimental study is carried out to find the optimal wavelength of the emitted light. To that end, under laboratory conditions, on the spectrophotometer model EMC-32PCS-UV, spectral measurements were performed on the absorption of emitted light by activated sludge microorganisms in the spectral range from 190 to 1100 nm wavelengths. Samples of activated sludge from the treatment plant in the city of Bukhara, Uzbekistan with different concentration values were utilized. When studying the absorption spectra of activated sludge microorganisms, it was experimentally established that the maximum absorption of light occurs in the infrared region of radiation, specifically at a wavelength of 940 nm. To select the optimal value of the radiation wavelength, the main criteria and requirements were theoretically determined. Based on these criteria, considering the scattering and absorption of light, the optical properties of the photodiodes used, it was concluded that the optimal value of the radiation wavelength is 940 nm. Using the results of the absorption spectra, the radiation wavelength was chosen. Graphical representations of the results of spectral studies with activated sludge microorganisms are given.

Removal of Diclofenac in Wastewater by Activated Sludge in Batch and Moving-bed Biofilm Reactor Experiments

This study aimed to determine the diclofenac removal in wastewater by sludge taken from an operating wastewater treatment facility. Laboratory experiments were conducted in two parts: batch and moving-bed biofilm reactor (MBBR) experiments. Results from batch experiments showed that 0.1–2 mg L-1 of diclofenac could be removed more than 80% within 72 h, and the removal efficiency reduced to less than 60% for higher concentrations. The increase in the removal rate from 0.00058 to 0.16527 mg L-1 h-1 was observed when the initial diclofenac concentration increased from 0.1 to 10 mg L-1, respectively. The average first-order rate constants of 24-h and 72-h degradation were calculated as 4.71 × 10-2 and 1.99 × 10-2 h-1, respectively. The removal of diclofenac by sludge was mainly from biodegradation by microorganisms in sludge, followed by the adsorption onto the sludge biomass. The addition of various metal ions in the studied range did not significantly increase the diclofenac removal; however, the addition of Ca2+, Co2+, Fe3+, Mn2+, and Zn2+ tended to increase both diclofenac removal rate and efficiency. This positive effect was reduced when the metal ion concentrations were increased up to 0.75 ppm. Lastly, results from an initial phase of continuous MBBR showed that sludge addition during the start-up also extended the diclofenac removal efficiency to one week compared with 3 days in the experiment without sludge addition. In conclusion, the findings show the capability of using activated sludge in diclofenac wastewater treatment by the traditional or alternative systems.

Horizontal gene transfer in activated sludge enhances microbial antimicrobial resistance and virulence

Activated sludge (AS) plays a vital role in removing organic pollutants and nutrients from wastewater. However, the risks posed by horizontal gene transfer (HGT) between bacteria in AS are still unclear. Here, a total of 478 high-quality non-redundant metagenome-assembled genomes (MAGs) were obtained. >50 % and 5 % of MAGs were involved in at least one HGT and recent HGT, respectively. Most of the transfers (82.4 %) of antimicrobial resistance genes (ARGs) occurred among the classes of Alphaproteobacteria and Gammaproteobacteria. The bacteria involved in the transfers of virulence factor genes (VFGs) mainly include Alphaproteobacteria (42.3 %), Bacteroidia (19.2 %), and Gammaproteobacteria (11.5 %). Moreover, the number of ARGs and VFGs in the classes of Alphaproteobacteria and Gammaproteobacteria was higher than that in other bacteria (P < 0.001). Mobile genetic elements were important contributors to ARGs and VFGs in AS bacteria. These results have implications for the management of antimicrobial resistance and virulence in activated sludge microorganisms.

Oxygen Uptake Rate as an Indicator of the Substrates Utilized by Candidatus Accumulibacter

Candidatus Accumulibacter belongs to phosphate-accumulating organisms (PAOs) which exhibit a cyclic metabolism and are capable of intracellular polyphosphate accumulation and their hydrolysis under feast-famine anaerobic-aerobic cycling. In consortia of activated sludge microorganisms, these bacteria are responsible for enhanced biological phosphorus removal (EBPR). The spectrum of the substrates used by Ca. Accumulibacter remains insufficiently studied. It was investigated by measuring the oxygen uptake rates (OUR) of Ca. Accumulibacter-enriched culture supplemented with 17 different organic substrates. The highest oxygen uptake rate values were observed in the presence of tryptone, volatile fatty acids (acetate, propionate, and butyrate), succinate, pyruvate, and amino acids (aspartate and glutamate). Phosphate dynamics in the medium under shifts from anaerobic to aerobic cultivation in batch experiments were studied for these compounds (except for tryptone). All tested substrates were shown to cause phosphate cycling (release in the anaerobic phase and uptake in the aerobic one), with OURs for the substrates correlating with the number of phosphates consumed during the aerobic phase. It was concluded that OUR may be used as an indicator of the monosubstrates used by Ca. Accumulibacter in the anaerobic/aerobic cycle. The possible pathways for substrate transport and metabolism by Ca. Accumulibacter are discussed using stoichiometric data and the results of metagenomic analysis.

Effects of lysed sludge reflux point on ultrasound lysis-cryptic growth in anaerobic/aerobic (A/O) wastewater treatment: Sludge reduction, microbial community, and metabolism

Ultrasonication allows sludge reduction to be performed in situ during wastewater treatment, and the reflux point of the lysed sludge affects this performance. This study investigated the effects of reflux point (anaerobic stage, carbon/nitrogen (C/N) lowest stage, and aerobic stage) on sludge lysis-cryptic growth in an anaerobic/aerobic reactor and variations in the sludge and microbial community. The best reflux point occurred at the lowest C/N ratio stage, and a 50.96% reduction in excess sludge was achieved. The reflux of the lysed sludge to the aerobic stage reduced nitrogen and phosphorus removal. The reflux of the lysed sludge decreased the average sludge size, reaching 29.2 μm when reflux to the aerobic stage. Scanning electron microscopy showed that the sludge surface was unaffected by the reflux point. The Fourier-transform infrared spectrometry and X-ray photoelectron spectroscopy results showed that the most prominent variation in the intensity of the sludge functional groups occurred when the reflux was at the lowest C/N stage. The amount of extracellular polymeric substances decreased the most during reflux to the anaerobic stage. The sludge microbial communities varied with the reflux point, and the dominant phyla during reflux to the anaerobic, lowest C/N, and aerobic stages were Bacteroidetes, Firmicutes, and Bacteroidetes, respectively. Furthermore, the reflux point did not alter the metabolic pathway of sludge microorganisms but increased the number of enzymes in metabolic pathways.