A new strategy for accelerating recovery of anaerobic granular sludge after low-temperature shock: In situ regulation of quorum sensing microorganisms embedded in polyvinyl alcohol sodium alginate.

Low-temperature could inhibit the performance of anaerobic granular sludge (AnGS). Quorum sensing (QS), as a communication mode between microorganisms, can effectively regulate AnGS. In this study, a kind of embedded particles (PVA/SA@Serratia) based on signal molecule secreting bacteria was prepared by microbial immobilization technology based on polyvinyl alcohol and sodium alginate to accelerate the recovery of AnGS system after low temperature. Low-temperature shock experiment verified the positive effect of PVA/SA@Serratia on restoring the COD removal rate and methanogenesis capacity of AnGS. Further analysis by metagenomics analysis showed that PVA/SA@Serratia stimulated higher QS activity and promoted the secretion of extracellular polymeric substance (EPS) in AnGS. The rapid construction of EPS protective layer effectively accelerated the establishment of a robust microbial community structure. PVA/SA@Serratia also enhanced multiple methanogenic pathways, including direct interspecies electron transfer. In conclusion, this study demonstrated that PVA/SA@Serratia could effectively strengthen AnGS after low-temperature shock.

Insights into the influence of polystyrene microplastics on the bio-degradation behavior of tetrabromobisphenol A in soil.

Soil contamination by emerging pollutants tetrabromobisphenol A (TBBPA) and microplastics has become a global environmental issue in recent years. However, little is known about the effect of microplastics on degradation of TBBPA in soil, especially aged microplastics. In this study, the effect of aged polystyrene (PS) microplastics on the degradation of TBBPA in soil and the mechanisms were investigated. The results suggested that the aged microplastics exhibited a stronger inhibitory effect on the degradation of TBBPA in soil than the pristine microplastics, and the degradation efficiency of TBBPA decreased by 21.57% at the aged microplastic content of 1%. This might be related to the higher TBBPA adsorption capacity of aged microplastics compared to pristine microplastics. Aged microplastics strongly altered TBBPA-contaminated soil properties, reduced oxidoreductase activity and affected microbial community composition. The decrease in soil oxidoreductase activity and relative abundance of functional microorganisms (e.g., Bacillus, Pseudarthrobacter and Sphingomonas) caused by aged microplastics interfered with metabolic pathways of TBBPA. This study indicated the importance the risk assessment and soil remediation for TBBPA-contaminated soil with aged microplastics.

Effect of high corn straw loads on short-chain fatty acid production in semi-continuous rumen reactor.

Ruminal microorganisms can efficiently hydrolyze biomass waste for short-chain fatty acid (SCFA) production. However, the continuous SCFA production by ruminal microorganisms at high loads is unclear. In this study, the effectiveness of a rumen semi-continuous reactor at high load for SCFA production was explored. Results showed that SCFA concentration reached 13.3 g/L at 8 % (w/v) corn straw load. The higher the corn straw load, the lower the volatile solid removal. Rumen microbial community composition changed significantly with increasing corn straw load. A significant decrease in bacterial diversity and abundance was observed at 8 % corn straw load. Some core genera such as Prevotella, Saccharofermentans, and Ruminococcus significantly increased. As corn straw loads increased, the expression of functional genes related to hydrolysis and acidogenesis gradually increased. Thus, the 8.0 % load is suitable for SCFA production. These findings provide new insights into high load fermentation of ruminal microorganisms.

Recent advances in ultrasound-Fenton/Fenton-like technology for degradation of aqueous organic pollutants.

Organic pollutants in water are a serious problem because of their widespread presence, harming the ecosystem and human health. Of the commonly used advanced oxidation processes, a hybrid of ultrasound and the Fenton/Fenton-like technology has received increasing attention in treatment of aqueous organic pollutants. This hybrid is effective in degradation of organic pollutants, but its application has not been summarised. Herein, first, the application and influencing factors of this hybrid technology for organic pollutants degradation are introduced. Second, the mechanism of its action is discussed. Third, the current challenges and future perspectives associated with this technology are proposed. This review provides valuable information regarding this technology, deepens the understanding of its mechanisms of organic pollutants degradation and provides a reference for its use in treatment of aquatic environments.

Rumen microbes, enzymes, metabolisms, and application in lignocellulosic waste conversion - A comprehensive review.

The rumen of ruminants is a natural anaerobic fermentation system that efficiently degrades lignocellulosic biomass and mainly depends on synergistic interactions between multiple microbes and their secreted enzymes. Ruminal microbes have been employed as biomass waste converters and are receiving increasing attention because of their degradation performance. To explore the application of ruminal microbes and their secreted enzymes in biomass waste, a comprehensive understanding of these processes is required. Based on the degradation capacity and mechanism of ruminal microbes and their secreted lignocellulose enzymes, this review concentrates on elucidating the main enzymatic strategies that ruminal microbes use for lignocellulose degradation, focusing mainly on polysaccharide metabolism-related gene loci and cellulosomes. Hydrolysis, acidification, methanogenesis, interspecific H2 transfer, and urea cycling in ruminal metabolism are also discussed. Finally, we review the research progress on the conversion of biomass waste into biofuels (bioethanol, biohydrogen, and biomethane) and value-added chemicals (organic acids) by ruminal microbes. This review aims to provide new ideas and methods for ruminal microbe and enzyme applications, biomass waste conversion, and global energy shortage alleviation.

Bioconversion of volatile fatty acids from organic wastes to produce high-value products by photosynthetic bacteria: A review.

Anaerobic fermentation of organic waste to produce volatile fatty acids (VFAs) production is a relatively mature technology. VFAs can be used as a cheap and readily available carbon source by photosynthetic bacteria (PSB) to produce high value-added products, which are widely used in various applications. To better enhance the VFAs obtained from organic wastes for PSB to produce high value-added products, a comprehensive review is needed, which is currently not available. This review systematically summarizes the current status of microbial proteins, H2, poly-ß-hydroxybutyrate (PHB), coenzyme Q10 (CoQ10), and 5-aminolevulinic acid (ALA) production by PSB utilizing VFAs as a carbon resource. Meanwhile, the metabolic pathways involved in the H2, PHB, CoQ10, and 5-ALA production by PSB were deeply explored. In addition, a systematic resource utilization pathway for PSB utilizing VFAs from anaerobic fermentation of organic wastes to produce high value-added products was proposed. Finally, the current challenges and priorities for future research were presented, such as the screening of efficient PSB strains, conducting large-scale experiments, high-value product separation, recovery, and purification, and the mining of metabolic pathways for the VFA utilization to generate high value-added products by PSB.

Response of rumen microorganisms to pH during anaerobic hydrolysis and acidogenesis of lignocellulose biomass.

Rumen microorganisms can efficiently degrade lignocellulosic wastes to produce volatile fatty acids (VFAs). pH is a key factor in controlling the type and yield of VFAs by affecting the microorganisms involved in rumen fermentation. However, the effects of different pH on rumen microbial diversity, communities, and mechanisms are unclear. In this study, the hydrolysis and acidogenesis of corn straw and diversity, communities, and mechanisms of rumen microorganisms were explored at different initial pHs. Results showed that the highest hemicellulose, cellulose, and lignin degradation efficiency of corn straw was 55.2 %, 38.3 %, and 7.01 %, respectively, and VFA concentration was 10.2 g/L at pH 7.0. Low pH decreased the bacterial diversity and increased the fungal diversity. Rumen bacteria and fungi had different responses to initial pHs, and the community structure of bacteria and fungi had obviously differences at the genus level. The core genera Succiniclasticum, Treponema, and Neocallimastix relative abundance at initial pH 7.0 samples were significantly higher than that at lower initial pHs, reaching 6.01 %, 1.61 %, and 5.35 %, respectively. The bacterial network was more complex than that of fungi. pH, acetic acid, and propionic acid were the main factors influencing the bacterial and fungal community structure. Low pH inhibited the expression of functional genes related to hydrolysis and acidogenesis, explaining the lower hydrolysis and acidogenesis efficiency. These findings will provide a better understanding for rumen fermentation to produce VFAs.

A global perspective on microbial risk factors in effluents of wastewater treatment plants.

Effective monitoring and management of microbial risk factors in wastewater treatment plants (WWTPs) effluents require a comprehensive investigation of these risks. A global survey on microbial risk factors in WWTP effluents could reveal important insights into their risk features. This study aims to explore the abundance and types of antibiotic resistance genes (ARGs), virulence factor genes (VFGs), the vector of ARG/VFG, and dominant pathogens in global WWTP effluents. We collected 113 metagenomes of WWTP effluents from the Sequence Read Archive of the National Center for Biotechnology Information and characterized the microbial risk factors. Our results showed that multidrug resistance was the dominant ARG type, while offensive virulence factors were the most abundant type of VFGs. The most dominant types of ARGs in the vector of plasmid and phage were both aminoglycoside resistance, which is concerning as aminoglycosides are often a last resort for treating multi-resistant infections. Acinetobacter baumannii was the most dominant pathogen, rather than Escherichia coli, and a weak negative correlation between Escherichia coli and two other dominant pathogens (Acinetobacter baumannii and Bacteroides uniformis) suggests that using Escherichia coli as a biological indicator for all pathogens in WWTP effluents may not be appropriate. The Getah virus was the most dominant virus found in global WWTP effluents. Our study presents a comprehensive global-scale investigation of microbial risk factors in WWTP effluents, providing valuable insights into the potential risks associated with WWTP effluents and contributing to the monitoring and control of these risks.

Adsorption behaviors and bioavailability of tetrabromobisphenol A in the presence of polystyrene microplastic in soil: Effect of microplastics aging.

Microplastics, a kind of emerging pollutant, have become a global environmental research hotspot in recent years due to its wide distribution in soil and its impact on soil ecosystems. However, little information is available on the interactions between microplastics and organic contaminants in soil, especially after microplastic aging. The impact of polystyrene (PS) microplastic aging on the sorption of tetrabromobisphenol A (TBBPA) in soil and the desorption characteristics of TBBPA-loaded microplastics in different environments were studied. The results showed a significant increase of 76.3% in adsorption capacity of TBBPA onto PS microplastics after aging for 96 h. Based on the results of characterization analysis and density functional theory (DFT) calculation, the mechanisms of TBBPA adsorption changed mainly from hydrophobic and π-π interactions on pristine PS microplastics to hydrogen bond and π-π interactions on aged PS microplastics. The presence of PS microplastics increased the TBBPA sorption capacity onto soil-PS microplastics system and significantly altered the distribution of TBBPA on soil particles and PS microplastics. The high TBBPA desorption over 50% from aged PS microplastics in simulated earthworm gut environment suggested that TBBPA contamination combined with PS microplastics might pose a higher risk to macroinvertebrates in soil. Overall, these findings contribute to the understanding of impact of PS microplastic aging in soil on the environmental behaviors of TBBPA, and provide valuable reference for evaluating the potential risk posed by the co-existence of microplastics with organic contaminants in soil ecosystems.

Metagenomic analysis reveals the efficient digestion mechanism of corn stover in Angus bull rumen: Microbial community succession, CAZyme composition and functional gene expression.

Ruminant rumen is a biological fermentation system that can efficiently degrade lignocellulosic biomass. The knowledge about mechanisms of efficient lignocellulose degradation with rumen microorganisms is still limited. In this study, composition and succession of bacteria and fungi, carbohydrate-active enzymes (CAZymes), and functional genes involved in hydrolysis and acidogenesis were revealed during fermentation in Angus bull rumen via metagenomic sequencing. Results showed that degradation efficiency of hemicellulose and cellulose reached 61.2% and 50.4% at 72 h fermentation, respectively. Main bacterial genera were composed of Prevotella, Butyrivibrio, Ruminococcus, Eubacterium, and Fibrobacter, and main fungal genera were composed of Piromyces, Neocallimastix, Anaeromyces, Aspergillus, and Orpinomyces. Principal coordinates analysis indicated that community structure of bacteria and fungi dynamically changed during 72 h fermentation. Bacterial networks with higher complexity had stronger stability than fungal networks. Most CAZyme families showed a significant decrease trend after 48 h fermentation. Functional genes related to hydrolysis decreased at 72 h, while functional genes involved in acidogenesis did not change significantly. These findings provide a in-depth understanding of mechanisms of lignocellulose degradation in Angus bull rumen, and may guide the construction and enrichment of rumen microorganisms in anaerobic fermentation of waste biomass.

A global perspective on compositions, risks, and ecological genesis of antibiotic resistance genes in biofilters of drinking water treatment plants.

Antibiotic resistance genes (ARGs) in biofilters of drinking water treatment plants (DWTPs) are regarded to be a remarkable potential health risk to human. A global survey on ARGs in biofilters may help evaluate their risk features as a whole. This study aims to explore the compositions, risks, and ecological genesis of ARGs in the biofilters of DWTPs. In total, 98 metagenomes of DWTP biofilters were collected from Sequence Read Archive  (SRA)  of National center for Biotechnology Information (NCBI), and the main ARG types were recognized, with multidrug, bacitracin, and beta-lactam as the first three types. Source water types (surface water vs. groundwater) were found to significantly influence antibiotic resistome, overpassing biofilter media and locations. Although ARG abundances of surface water biofilters were approximately five times higher than that of groundwater biofilters, the risk pattern of ARGs was highly similar between surface water biofilters and groundwater biofilters, and up to 99.61% of the ARGs on average belong to the least risk and unassessed ranks, and only 0.23% the highest risk rank. Monobactam biosynthesis pathway and prodigiosin biosynthesis pathway, two antibiotics biosynthesis pathways, were observed to be positively correlated with several ARG types and total ARG abundance in samples of surface water and groundwater biofilters, respectively, suggesting their potential roles in ecological genesis of ARGs. Overall, the results of this study would deepen our understanding of the ARG risks in biofilters of DWTPs and shed light on their ecological genesis inside.

Polysaccharide from fermented mycelium of Inonotus obliquus attenuates the ulcerative colitis and adjusts the gut microbiota in mice.

Ulcerative colitis (UC) is a disease characterized by chronic inflammation of the colon. Polysaccharides not only have biological activities but also can regulate gut microbiota to alleviate the symptoms of UC. In this study, polysaccharide extracted from mycelium of Inonotus obliquus (IOP) was prescribed to treat UC induced by dextran sodium sulfate (DSS) in mice. Compared to model control group (MC), IOP-Low, IOP-Medium and IOP-High (IOP-L, IOP-M and IOP-H) treatment groups increased the body weight rate by 6.0%-9.6%, colon length by 8.57%-25.14% and superoxide dismutase (SOD) activity by 53.8-110.4 U/mg, while decreased the malondialdehyde (MDA) content by 37.4%-64.8%, myeloperoxidase (MPO) activity by 29.0%-46.9%, and the concentration of nitric oxide (NO) by 24.8-35.6 µmol/L. IOP treatment also promoted the secretion of interleukin (IL)-10 but suppressed those of interleukin (IL)-6, interleukin (IL)-1ß and tumor necrosis factor (TNF)-α. Simultaneously, analysis of high-throughput sequencing indicated that IOP reduced the ratio of Firmicutes to Bacteroidetes (F/B) at phylum level, and increased the relative abundance of Bacteroides and Lactobacillus at genus level. In brief, IOP may be a promising alternative medicine for UC remedy by regulating the anti-inflammatory level, the anti-oxidative ability and the gut microbiota composition.

Risk assessment of three sheep stocking modes via identification of bacterial genomes carrying antibiotic resistance genes and virulence factor genes.

In order to protect the prairie ecological environment, intensive farming has become a prevalent method of sheep stocking. However, the link between captivity stocking mode and ecological risk of sheep feces is still poorly understood. In this study, metagenomics was used to identify the environmental risk of sheep feces among three stocking modes. Our results showed that captivity mode (C) elevated antibiotic resistance in feces, with the abundance of antibiotic resistance genes (ARGs) (5.381 copies/cell) higher than that of half-pen stocking (Fh) (1.093 copies/cell) and grazing mode (Fr) (0.315 copies/cell) (Duncan's test, P < 0.05). Virulence factor genes (VFGs) analysis showed offensive virulence factors had the highest abundance in captivity feces (C: 3.826 copies/cell, Fh: 0.342 copies/cell, Fr: 0.163 copies/cell) (Duncan's test, P < 0.05). 15 metagenome-assembled genomes (MAGs) were identified as potential pathogenic antibiotic resistant bacteria (PARB) and revealed that Escherichia, Klebsiella may be the main host of ARGs and VFGs in sheep feces. Furthermore, the minimal inhibition concentrations (MIC) of tetracycline of E. coli in the captivity feces was 8.6 times and 4.7 times than that of grazing and half-pen stocking samples, respectively. The Non-metric multidimensional scaling (NMDS) revealed that high stocking density leads to feces causing increased harm to the environment. Although feces from sheep raised in captivity and half-pen stocking modes are easier to collect, they are more harmful to the environment and aerobic composting should be done before their application to farmland. This work provides a guideline for better control of the environmental risk of sheep feces from different stocking modes.

Combining high pressure homogenization with free nitrous acid pretreatment to improve anaerobic digestion of sewage sludge.

Single pretreatment of sewage sludge, either physical, chemical or biological, has its own drawbacks in term of poor sanitization, energy intensity and high operational and capital cost. To tackle these drawbacks, combined high pressure homogenization (HPH) and free nitrous acid (FNA) pretreatment for sludge solubilization and further biodegradation in anaerobic digestion was investigated. Synergistic effect of combined HPH (40 MPa) and FNA (2.49 mg/L) pretreatment (HPH-FNA) for improving anaerobic digestion was evaluated, and its effect on archaeal and bacterial community structure was analyzed. Compared with single HPH and FNA pretreatments, HPH-FNA pretreatment efficiently solubilized wasted activated sludge (WAS), subsequently improved anaerobic digestion. Cumulative biogas production from sewage sludge pretreated with HPH-FNA was 154%, 108% and 284% more than that with single pretreatment of FNA, HPH and raw sludge, respectively. In addition, volumetric biogas production of combined pretreatment system (815 ml) was more than the sum from single pretreatment (710 ml). Methane content in biogas for raw sludge, FNA, HPH and HPH-FNA pretreated sludge was 45%, 51%, 55% and 65%, respectively. Illumina MiSeq sequencing analysis revealed that HPH-FNA pretreatment promoted bacterial growth of phyla Bacteroidetes, Firmicutes and Synergistetes and archaeal genera Methanospirillum and Methanosaeta. Overall, combined HPH-FNA pretreatment of sewage sludge, prior to anaerobic digestion, is an environmentally-friendly and potentially economic technology.

Toxicological effects, environmental behaviors and remediation technologies of herbicide atrazine in soil and sediment: A comprehensive review.

Atrazine has become one of the most popular applied triazine herbicides in the world due to its high herbicidal efficiency and low price. With its large-dosage and long-term use on a global scale, atrazine can cause widespread and persistent contamination of soil and sediment. This review systematically evaluates the toxicological effects, environmental risks, environmental behaviors (adsorption, transport and transformation, and bioaccumulation) of atrazine, and the remediation technologies of atrazine-contaminated soil and sediment. For the adsorption behavior of atrazine on soil and sediment, the organic matter content plays an extremely important role in the adsorption process. Various models and equations such as the multi-media fugacity model and solute transport model are used to analyze the migration and transformation process of atrazine in soil and sediment. It is worth noting that certain transformation products of atrazine in the environment even have stronger toxicity and mobility than its parent. Among various remediation technologies, the combination of microbial remediation and phytoremediation for atrazine-contaminated soil and sediment has wide application prospects. Although other remediation technologies such as advanced oxidation processes (AOPs) can also efficiently remove atrazine from soil, some potential problems still need to be further clarified. Finally, some related challenges and prospects are proposed.

A critical review on interaction of microplastics with organic contaminants in soil and their ecological risks on soil organisms.

The pollution of microplastics (MPs) in soil has become a global environmental problem. Due to high sorption capacity and persistence in environment, the MPs exhibit combined effects with organic pollutants in soil, thereby posing a potential risk to soil ecology and human health. However, limited reviews are available on this subject. Therefore, in response to this issue, this review provides an in-depth account of interaction of MPs with organic contaminants in soil and the combined risks to soil environment. The sorption of organic contaminants onto MPs is mainly through hydrophobic and π-π interactions, hydrogen bonding, pore filling and electrostatic and van der Waals forces. The intrinsic characteristics of MPs, organic contaminants and soil are the key factors influencing the sorption of organic pollutants onto MPs. Importantly, the presence of MPs changes the sorption, degradation and transport behaviors of organic contaminants in soil, and affects the toxic effects of organic contaminants on soil organisms including animals, plants and soil microorganisms through synergistic or antagonistic effects. Source control, policy implementation and plastic removal are the main preventive and control measures to reduce soil MPs pollution. Finally, priorities for future research are proposed, such as field investigations of co-pollution, contribution of plastisphere to organic contaminant degradation, and mechanisms of MPs effects on organic contaminant toxicity.

Long-term rumen microorganism fermentation of corn stover in vitro for volatile fatty acid production.

Rumen microorganisms have the ability to efficiently hydrolyze and acidify lignocellulosic biomass. The effectiveness of long-term rumen microorganism fermentation of lignocellulose in vitro for producing volatile fatty acids (VFAs) is unclear. The feasibility of long-term rumen microorganism fermentation of lignocelluose was evaluated in this study, and a stable VFA production was successfully realized for 120 d. Results showed that VFA concentration reached to 5.32-8.48 g/L during long-term fermentation. Hydrolysis efficiency of hemicellulose and cellulose reached 36.5%-52.2% and 29.4%-38.4%, respectively. A stable bacterial community was mainly composed of Prevotella, Rikenellaceae_RC9_gut_group, Ruminococcus, and Succiniclasticum. VFA accumulation led to a pH decrease, which caused the change of bacterial community structure. Functional prediction showed that the functional genes related to hydrolysis and acidogenesis of corn stover were highly expressed during long-term fermentation. The successful long-term rumen fermentation to produce VFAs is of great significance for the practical application of rumen microorganisms.

Improvement of Direct Interspecies Electron Transfer via Adding Conductive Materials in Anaerobic Digestion: Mechanisms, Performances, and Challenges.

Anaerobic digestion is an effective and sustainable technology for resource utilization of organic wastes. Recently, adding conductive materials in anaerobic digestion to promote direct interspecies electron transfer (DIET) has become a hot topic, which enhances the syntrophic conversion of various organics to methane. This review comprehensively summarizes the recent findings of DIET mechanisms with different mediating ways. Meanwhile, the influence of DIET on anaerobic digestion performance and the underlying mechanisms of how DIET mediated by conductive materials influences the lag phase, methane production, and system stability are systematically explored. Furthermore, current challenges such as the unclear biological mechanisms, influences of non-DIET mechanisms, limitations of organic matters syntrophically oxidized by way of DIET, and problems in practical application of DIET mediated by conductive materials are discussed in detail. Finally, the future research directions for practical application of DIET are outlined.

High salinity slowed organic acid production from acidogenic fermentation of kitchen wastewater by shaping functional bacterial community.

The high salinity of kitchen wastewater might have adverse effects on the production of short-chain fatty acids (SCFAs) in anaerobic fermentation. The effects and mechanisms of salinity on SCFA production in the anaerobic fermentation of kitchen wastewater were studied by varying the salt concentration, as follows: 0 g/L (S0), 2 g/L (S2), 6 g/L (S6), 10 g/L (S10), 15 g/L (S15), and 20 g/L (S20). Experimental results showed that hypersaline conditions (>10 g NaCl/L) accelerated the release of soluble proteins at the initial stage of anaerobic fermentation. They also significantly prohibited the hydrolysis and degradation of soluble proteins and carbohydrates. Compared with low salinity tests, the SCFA concentrations under hypersaline conditions (>10 g NaCl/L) only reached approximately 43% of the highest concentration on day 10, although the SCFA concentrations in all tests were very close on day 10 (14 g COD/L). High salinity delayed the production of n-butyric acid but did not change the composition of the total SCFAs. High salinity enriched Enterococcus and Bifidobacterium, the relative abundance levels of which reached 27.57% and 49.71%, respectively, before the depletion of substrate. High salinity showed a negative correlation with the relative abundance of the genera Clostridium_sensu_stricto_1, Prevotella and unclassified_f_Oscillospiraceae which are responsible for SCFA production. This study provided a theoretical basis for the fficient utilization of kitchen wastewater.

Siderophores provoke extracellular superoxide production by Arthrobacter strains during carbon sources-level fluctuation.

Superoxide and other reactive oxygen species (ROS) shape microbial communities and drive the transformation of metals and inorganic/organic matter. Taxonomically diverse bacteria and phytoplankton produce extracellular superoxide during laboratory cultivation. Understanding the physiological reasons for extracellular superoxide production by aerobes in the environment is a crucial question yet not fully solved. Here, we showed that iron-starving Arthrobacter sp. QXT-31 (A. QXT-31) secreted a type of siderophore [deferoxamine (DFO)], which provoked extracellular superoxide production by A. QXT-31 during carbon sources-level fluctuation. Several other siderophores also demonstrated similar effects to A. QXT-31. RNA-Seq data hinted that DFO stripped iron from iron-bearing proteins in electron transfer chain (ETC) of metabolically active A. QXT-31, resulting in electron leakage from the electron-rich (resulting from carbon sources metabolism by A. QXT-31) ETC and superoxide production. Considering that most aerobes secrete siderophore(s) and undergo carbon sources-level fluctuation, the superoxide-generation pathway is likely a common pathway by which aerobes produce extracellular superoxide in the environment, thus influencing the microbial community and cycling of elements. Our results pointed that the ubiquitous siderophore might be the potential driving force for the microbial generation of superoxide and other ROS and revealed the important role of iron physiology in microbial ROS generation.