Extracellular Polymeric Substances Production Research Articles

Role of biochar addition to improve anaerobic membrane bioreactors to resist oil stress in synthetic food wastewater treatment

This study investigated biochar addition to improve the performance and stability of anaerobic membrane bioreactor (AnMBR) to resist different oil contents in food wastewater treatment. Results show that an increase in oil content in synthetic food wastewater from 2 to 4 g/L of oleate-Na aggravated AnMBR performance in terms of methane (CH4) production and membrane fouling as showed by an obvious increase in transmembrane pressure (TMP). The reduced performance was caused by the accumulation of volatile fatty acids (VFAs) to inhibit the growth and activity of Methanobacterium in response to increased oil content. Nevertheless, biochar addition improved AnMBR resistance to against oil increase. The dominance of Methanobacterium and their potential mutualism with genus T78 were enhanced by biochar to sustain CH4 yield. Moreover, biochar reduced the production of extracellular polymeric substance (EPS) to alleviate membrane fouling.

Microbial community and extracellular polymeric substance dynamics in arid–zone temporary pan ecosystems

Microbial extracellular polymeric substances (EPS) are an important component in sediment ecology. However, most research is highly skewed towards the northern hemisphere and in more permanent systems. This paper investigates EPS (i.e., carbohydrates and proteins) dynamics in arid Austral zone temporary pans sediments. Colorimetric methods and sequence–based metagenomics techniques were employed in a series of small temporary pan ecosystems characterised by alternating wet and dry hydroperiods. Microbial community patterns of distribution were evaluated between seasons (hot–wet and cool–dry) and across depths (and inferred inundation period) based on estimated elevation. Carbohydrates generally occurred in relatively higher proportions than proteins; the carbohydrate:protein ratio was 2.8:1 and 1.6:1 for the dry and wet season respectively, suggesting that EPS found in these systems was largely diatom produced. The wet– hydroperiods (Carbohydrate mean 102 μg g−1; Protein mean 65 μg g−1) supported more EPS production as compared to the dry– hydroperiods (Carbohydrate mean 73 μg g−1; Protein mean 26 μg g−1). A total of 15,042 Unique Amplicon Sequence Variants (ASVs) were allocated to 51 bacterial phyla and 1127 genera. The most abundant genera had commonality in high temperature tolerance, with Firmicutes, Actinobacteria and Proteobacteria in high abundances. Microbial communities were more distinct between seasons compared to within seasons which further suggested that the observed metagenome functions could be seasonally driven. This study's findings implied that there were high levels of denitrification by mostly nitric oxide reductase and nitrite reductase enzymes. EPS production was high in the hot–wet season as compared to relatively lower rates of nitrification in the cool–dry season by ammonia monooxygenases. Both EPS quantities and metagenome functions were highly associated with availability of water, with high rates being mainly associated with wet– hydroperiods compared to dry– hydroperiods. These data suggest that extended dry periods threaten microbially mediated processes in temporary wetlands, with implications to loss of biodiversity by desiccation.

Quantitative image analysis for assessing extracellular polymeric substances in activated sludge under atrazine exposure

Extracellular polymeric substances (EPS) play a vital role in biological wastewater treatment systems. This study investigates the impact of herbicide atrazine (ATZ) on the overall performance, EPS yield, composition, and sludge morphology in an activated sludge (AS) system operated in a sequencing batch reactor (SBR). Since conventional methods for analyzing EPS are time-consuming and releases residues, a new approach was developed in this work to evaluate the EPS fractions and components, based on the morphological characterization of the biomass using quantitative image analysis (QIA) technique coupled with multivariate statistics.Results showed that exposure to ATZ inhibit biomass activity in terms of organic matter (COD) and nitrogen removal. Moreover, both tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) increased under ATZ, indicating that microorganisms release EPS as a defense mechanism against environmental changes. The PN/PS ratio also increases, indicating likely increased hydrophobicity in ATZ phases. Furthermore, ATZ phases exhibit a predominance of large aggregates compared to intermediate and small ones, indicating a change in aggregate morphological structure associated with EPS production. The new approach using QIA coupled with partial least squares (PLS) modeling provides accurate predictions of EPS content. The increase in TB-EPS is closely related to the rise of large aggregates in phases exposed to higher ATZ concentrations. The PLS models demonstrate high accuracy for EPS prediction (coefficients of determination, R2 above 0.86), showcasing the feasibility of using QIA for EPS assessment in AS systems. This approach offers significant potential for regular process monitoring and management, providing a more environmentally friendly methodology by eliminating the need for chemical usage.

The cumulative impact of temperature and nitrogen availability on the potential nitrogen fixation and extracellular polymeric substances secretion by Dolichospermum

Nitrogen-fixing cyanobacteria not only cause severe blooms but also play an important role in the nitrogen input processes of lakes. The production of extracellular polymeric substances (EPS) and the ability to fix nitrogen from the atmosphere provide nitrogen-fixing cyanobacteria with a competitive advantage over other organisms. Temperature and nitrogen availability are key environmental factors in regulating the growth of cyanobacteria. In this study, Dolichospermum (formerly known as Anabaena) was cultivated at three different temperatures (10°C, 20°C, and 30°C) to examine the impact of temperature and nitrogen availability on nitrogen fixation capacity and the release of EPS. Initially, confocal laser scanning microscopy (CLSM) and the quantification of heterocysts at different temperatures revealed that lower temperatures (10°C) hindered the differentiation of heterocysts under nitrogen-deprived conditions. Additionally, while heterocysts inhibited the photosynthetic activity of Dolichospermum, the secretion of EPS was notably affected by nitrogen limitation, particularly at 30°C. Finally, real-time quantitative polymerase chain reaction (qPCR) was used to measure the expression of nitrogen-utilizing genes (ntcA and nifH) and EPS synthesis-related genes (wzb and wzc). The results indicated that under nitrogen-deprived conditions, the expression of each gene was upregulated, and there was a significant correlation between the upregulation of nitrogen-utilizing and EPS synthesis genes (P < 0.05). Our findings suggested that Dolichospermum responded to temperature variation by affecting the formation of heterocysts, impacting its potential nitrogen fixation capacity. Furthermore, the quantity of EPS released was more influenced by nitrogen availability than temperature. This research enhances our comprehension of interconnections between nitrogen deprivation and EPS production under the different temperatures.

Varying removal of emerging organic contaminants in drinking water treatment residue-based biofilter systems: Influence of hydraulic loading rates on microbiology

Drinking water treatment residue (DWTR)-based biofilter is an environment-friendly and cost-effective technology for eliminating nutrients and emerging organic contaminants (EOCs) from natural water. This study explored the removal efficiencies of five trace-level EOCs and the associated microbial response of biofilm in two identical DWTR-based biofilters under varying hydraulic loading rate (HLR) conditions over six months of operation. The results showed that the most recalcitrant EOC (carbamazepine) was mainly removed by DWTR adsorption (7.8–11.0 ng g−1), while the response of the biofilm community to HLR significantly affected the degradation of 17α-ethinylestradiol, sulfamethoxazole, roxithromycin, and sulfathiazole (P < 0.001). A higher HLR significantly stimulated the production of extracellular polymeric substance and increased the stochasticity and diversity of community assembly, improving denitrification and the removal efficiency of DWTR-based biofilm for biodegradable roxithromycin (80 %) and sulfamethoxazole (76 %). In contrast, lower HLR led to carbon limitation and exerted substrate selection pressures, resulting in more deterministic and stable community assembly, with keystone species such as Bacillus enriched in EOC degradation-related functional genes. Moreover, co-occurrence network analysis revealed a simpler but more intensive interaction network among EOC degraders under the lower HLR condition. This interaction network facilitated the co-metabolism of influent organic carbon and the removal of more recalcitrant contaminants, 17α-ethinylestradiol (48 %) and sulfathiazole (37 %). This study offered a novel insight into biofilter management from a hybrid perspective of environmental microbiology and engineering science, highlighting the dynamic adaptation of microbes to specific EOC degradation under varying HLR conditions.

Bioremediation of heavy metals-contaminated sites by microbial extracellular polymeric substances – A critical view

Heavy metal contamination in the atmosphere poses substantial threats to ecosystems. Traditional methods for the removal of HM-contaminated sites, such as physical and chemical treatments, often have limitations in terms of efficiency, cost, and potential negative impacts on the environment. As a result, there is growing interest in exploring alternative and sustainable approaches, such as bioremediation. Bioremediation is a promising technology that utilizes the inherent abilities of microorganisms to degrade or transform pollutants into less harmful forms. Microbial exopolysaccharides have gained attention as potential tools in bioremediation strategies due to their unique physicochemical properties and their ability to interact with heavy metals. This work aims to provide an overview of the role of microbial extracellular polymeric substances (EPS) in the bioremediation of HM-contaminated sites. First, the mechanisms by which EPS facilitates heavy metal sequestration are discussed. EPS can form metal-complexing agents, such as chelators and ion-binding sites, which enhance the immobilization and precipitation of heavy metals, reducing their bioavailability and toxicity. EPS can act as a protective barrier, shielding microorganisms from heavy metal stress and facilitating their survival and growth. Additionally, EPS production can be induced or enhanced in response to heavy metal exposure, potentially increasing the efficiency of bioremediation processes.

Membrane fouling during the harvesting of microalgae using static microfiltration

A challenge in the filtration of algal suspensions is the reduction in filtrate flow rate with time, associated with membrane fouling and filtercake formation. These two fouling mechanisms may occur simultaneously and have different effects on the scaling relationship of filtration rate with time, making quantitative predictions of the filtration behaviour complex. The foulants include suspended microalgal cells and their presumed Extracellular Polymeric Substances (EPS). To investigate this problem, we perform static microfiltration experiments to harvest oil-rich marine microalgae Nannochloropsis oculata. Batch filtration experiments of microalgal suspensions using glass-fibre membranes are conducted under filtration pressures varying between 0.5 and 200 kPa. Here we investigate the relative importance of potential fouling mechanisms, including pore plugging, entrance blocking, and filtercake formation. We examine variations in filtrate flux, using a novel approach involving numerical differentiation of the filtrate volume to identify how its scaling relationship with time and compare with the traditional root-time behaviour. These results are analysed with reference to the blocking filtration laws to determine the potential fouling mechanism. Rheology tests of both filtration feeds and filtrate are performed, and both optical and scanning electron microscopy are used to observe the filtercake. Our results show a significant drop in the filtrate flux after a spurt loss phase under pressure. The scaling analysis demonstrates a power law relationship between cumulative filtrate volume and time in the post-spurt phase. We show here, for the first time, that the scaling exponent varies with time, approaching a value close to 0.5 (i.e. the root-time behaviour) after a long period of time. Using the analysis, we conclude that the filtration process can be divided into various stages; a spurt loss phase when the pressure is initially applied, followed by a stage in which both internal and external filtercake formation occurs, and a final stage which is dominated by external filtercake formation (at which point root-time behaviour is observed). Furthermore, our findings suggest that fresh microalgal suspensions experienced a larger spurt loss compared to aged suspensions in microfiltration. This difference may be attributed to the limited production of EPS and microalgal debris during shorter cultivation periods. The microscopic observations reveal the invasion of microalgal cells into the membrane, which verifies the formation of an internal filtercake suggested by our scaling analysis. The contamination of the membrane increases with higher filtration pressures. Finally, we demonstrate that Fe3+ coagulant can be used to increase the microalgal particulate size before filtration, resulting in a much higher filtrate flux compared to uncoagulated suspensions. This result provides opportunities to reduce membrane contamination to a negligible level.

Sustainable production of extracellular polymeric substances and iron or copper complex from glutinous rice processing wastewater

Essential trace minerals play vital roles in maintaining human and animal health. However, an overdose of the existing inorganic trace minerals is prone to induce detrimental effects that outweigh positive benefits. In this study, an extracellular polymeric substances (EPS)-producing bacterium, identified as Bacillus licheniformis CCTCC M2020298, was isolated from marine using glutinous rice processing wastewater as enrichment medium. The EPS yield of Bacillus licheniformis CCTCC M2020298 could reach 8.62 g/L by using glutinous rice-processing wastewater containing medium. Furthermore, the potential of the EPS as a carrier for synthesizing EPS-iron (Fe) and EPS-copper (Cu) complex was explored. The results showed that the optimum condition for the synthesis EPS-Fe were the reaction temperature 70°C, pH 8.5–9.0 and mass ratio of EPS to trisodium citrate 2:1. The iron content of EPS-Fe reached 77.4 mg/g. Under the same condition, the copper content of EPS-Cu reached 90.7 mg/g. The elemental composition, functional groups and valence state of the mineral elements of EPS-Fe and EPS-Cu were well characterized. The EPS-Fe and EPS-Cu exhibited antioxidant activity in scavenging ·OH, DPPH and ·O2− free radicals, thereby leading to reduced oxidative stress and apoptosis levels in human colonic epithelial cells in vitro. They also inhibited the proliferation of mouse hepatocellular carcinoma H22 and the growth of intestinal pathogens in vitro. This study provided an effective avenue for EPS production from glutinous rice processing wastewater and proved the potential of EPS-Fe and EPS-Cu complexes as a new-type comprehensive essential trace mineral supplement.

The effect of cadmium on a semi-self-sustaining microalgal-bacterial granular sludge process for wastewater treatment

Microalgal-bacterial granular sludge (MBGS) is an innovative and sustainable technology for wastewater treatment. This research investigated the effect of Cd2+ on the performance and characteristics of such semi-self-sustaining MBGS reactors (with additional intermittent aeration). After 30 days of exposure to 0, 2, and 10 mg·l−1 Cd2+, the three reactors consistently exhibited robust removal efficiencies, achieving 89 % removal for chemical oxygen demand (COD) and 99 % removal for NH4+-N. However, the MBGS in control exhibited better performance in T-N removal compared to MBGS exposed to 10 mg·l−1 Cd2+ (especially during the light phase), indicating a potential impact of Cd2+ on the N-assimilation process of microalgae. The high concentration of Cd2+ also negatively affected the MBGS structure and increased the production of extracellular polymeric substances. Further investigations of the dissolved oxygen profile were conducted under phototrophic conditions, the maximum oxygen production rate contributed to microalgae in the control group was 20.9 % and 46.4 % higher than that of groups with 2 and 10 mg·l−1 Cd2+. The results show that Cd2+ mainly impacts the microalgae in the outer layer of the structure, and additional aeration helps compensate for the decrease in oxygen production caused by Cd2+ in MBGS, thereby making the system more stable in removing COD and NH4+-N.

Regulation of quorum sensing activities by the stringent response gene rsh in sphingomonads is species-specific and culture condition dependent.

Stringent response and quorum sensing (QS) are two essential mechanisms that control bacterial global metabolism for better survival. Sphingomonads are a clade of bacteria that survive successfully in diverse ecosystems. In silico survey indicated that 36 out of 79 investigated sphingomonads strains contained more than one luxI homolog, the gene responsible for the biosynthesis of QS signal acyl homoserine lactones (AHLs). Investigation of the regulatory effects of the stringent response gene rsh on QS related bioactivities were carried out using rsh mutants of Sphingobium japonicum UT26 and Sphingobium sp. SYK-6, both had three luxI homologs. Results indicated that deletion of rsh upregulated the overall production of AHLs and extracellular polymeric substances (EPS) in both UT26 and SYK-6 in rich medium, but affected expressions of these luxI/luxR homologs in different ways. In the poor medium (1% LB), rsh mutant of SYK-6 significantly lost AHLs production in broth cultivation but not in biofilm cultivation. The regulatory effects of rsh on QS activities were growth phase dependent in UT26 and culture condition dependent in SYK-6. Our results demonstrated the negative regulatory effect of rsh on QS activities in sphingomonads, which were very different from the positive effect found in sphingomonads containing only one luxI/R circuit. This study extends the current knowledge on the intricate networks between stringent response and QS system in sphingomonads, which would help to understand their survival advantage.

Development of a flow-cell bioreactor for immobilized sulfidogenic sludge characterization using electrochemical H2S microsensors

The sulfate-reduction process plays a crucial role in the biological valorization of SOx gases. However, a complete understanding of the sulfidogenic process in bioreactors is limited by the lack of technologies for characterizing the sulfate-reducing activity of immobilized biomass. In this work, we propose a flow-cell bioreactor (FCB) for characterizing sulfate-reducing biomass using H2S microsensors to monitor H2S production in real-time within a biofilm. To replace natural immobilization through extracellular polymeric substance production, sulfidogenic sludge was artificially immobilized using polymers. Physical and sulfate-reducing activity studies were performed to select a polymer-biomass matrix that maintained sulfate-reducing activity of biomass while providing strong microbial retention and mechanical strength. Several operational conditions of the sulfidogenic reactor allowed to obtain a H2S profiles under different inlet sulfate loads and, additionally, 3D mapping was assessed in order to perform a hydraulic characterization. Besides, the effects of artificial immobilization on biodiversity were investigated through the characterization of microbial communities. This study demonstrated the appropriateness of immobilized-biomass for characterization of sulfidogenic biomass in FCB using H2S electrochemical microsensors, and beneficial microbiological communities shifts as well as enrichment of sulfate-reducing bacteria have been confirmed.

Beyond feast and famine: Cultivating hydrodynamic oxygenic photogranules with better performances under permanent feast regime

Oxygenic photogranules (OPGs) are currently obtained in permanent famine or cyclic feast-famine regimes. Whether photogranulation occurs under a permanent feast regime and how these regimes impact OPGs are unknown. Herein, the three regimes, each applied in two replicate hydrodynamic reactors, were established by different feeding frequencies. Results showed that OPGs were successfully cultivated in all regimes after 24–36 days of photogranulation phases with similar microbial community functions, including filamentous gliding, extracellular polymeric substances production, and carbon/nitrogen metabolism. The OPGs were then operated under the same sequencing batch mode and all achieved efficient removal of chemical oxygen demand (>91 %), ammonium (>96 %), and total nitrogen (>76 %) after different adaptation periods (19–41 days). Notably, the permanent feast regime obtained OPGs with the best physicochemical properties, the shortest adaptation period, and the lowest effluent turbidity, thus representing a novel means of hydrodynamic cultivating OPGs with better performances for sustainable wastewater treatment.

Roles of extracellular polymeric substances in arsenic accumulation and detoxification by cell wall intact and mutant strains of Chlamydomonas reinhardtii

Arsenic (As) pollution seriously threatens human and ecological health. Microalgal cell wall and extracellular polymeric substances (EPS) are known to interact with As, but their roles in the As resistance, accumulation and speciation in microalgae remain unclear. Here, we used two strains of Chlamydomonas reinhardtii, namely CC-125 (wild type) and CC-503 (cell wall-deficient mutant), to examine the algal growth, EPS synthesis, As adsorption, absorption and transformation under 10–1000 µg/L As(III) and As(V) treatments for 96 h. In both strains, the As absorption increased after the EPS removal, but the growth, As adsorption, and transformation of C. reinhardtii declined. The CC-125 strain was more tolerant to As stress and more efficient in EPS production, As accumulation, and redox transformation than CC-503, irrespective of EPS presence or absence. Three-dimension excitation-emission matrix (3D-EEM) and attenuated total reflectance infrared spectroscopy (ATR-IR) analyses showed that As was bound with functional groups in the EPS and cell wall, such as -COOH, -NH and -OH in proteins, polysaccharides and amino acids. Together, this study demonstrated that EPS and cell wall acted as barriers to lower the As uptake by C. reinhardtii. However, the cell wall mutant strain was more susceptible to As toxicity due to lower EPS induction and higher As absorption.

Exploring Non-Thermal Plasma and UV Radiation as Biofilm Control Strategies against Foodborne Filamentous Fungal Contaminants.

In recent years, non-thermal plasma (NTP) has emerged as a promising tool for decontamination and disinfection within the food industry. Given the increasing resistance of microbial biofilms to conventional disinfectants and their adverse environmental effects, this method has significant potential for eliminating biofilm formation or mitigating the metabolic activity of grown biofilms. A comparative study was conducted evaluating the efficacy of UV radiation and NTP in eradicating mature biofilms of four common foodborne filamentous fungal contaminants: Alternaria alternata, Aspergillus niger, Fusarium culmorum, and Fusarium graminearum. The findings reveal that while UV radiation exhibits variable efficacy depending on the duration of exposure and fungal species, NTP induces substantial morphological alterations in biofilms, disrupting hyphae, and reducing extracellular polymeric substance production, particularly in A. alternata and F. culmorum. Notably, scanning electron microscopy analysis demonstrates significant disruption of the hyphae in NTP-treated biofilms, indicating its ability to penetrate the biofilm matrix, which is a promising outcome for biofilm eradication strategies. The use of NTP could offer a more environmentally friendly and potentially more effective alternative to traditional disinfection methods.

Delving into the Impacts of Different Easily Degradable Carbon Sources on the Degradation Characteristics of 2,4,6-Trichlorophenol and Microbial Community Properties

In chlorophenol wastewater treatment, adding easily degradable carbon sources, such as methanol, ethanol, sodium acetate, and sodium propionate, significantly improves the chlorophenol removal efficiency. This study systematically compares these conventional carbon sources in different sequencing batch reactors to understand their specific effects on both 2,4,6-trichlorophenol (2,4,6-TCP) degradation efficiency and microbial abundance. In a 35-day experiment, as a carbon source, ethanol exhibited a lower 2,4,6-TCP degradation concentration (77.56 mg/L) than those of methanol, sodium acetate, and sodium propionate, which achieved higher degradation concentrations: 123.89 mg/L, 170.96 mg/L, and 151.79 mg/L, respectively. As a carbon source, sodium acetate enhanced extracellular polymeric substance production (200.80 mg/g·VSS) by microorganisms, providing protection against the toxicity of chlorophenol and resulting in a higher 2,4,6-TCP removal concentration. Metagenomics identified crucial metabolic genes, including PcpA, chqB, Mal-r, pcaI, pcaF, and fadA. The abundance of genera containing the chqB gene correlated positively with the metabolic capacity for 2,4,6-TCP. Moreover, small molecular carbon sources such as methanol, sodium acetate, and sodium propionate promoted the enrichment of genera with functional genes.

Bioprospecting of Chlamydomonas reinhardtii for boosting biofuel-related products production based on novel aggregation-induced emission active extracellular polymeric substances nanoprobes

Biofuel production from microalgae has been greatly restricted by low biomass productivity and long-term photosynthetic efficacy. Here, a novel strategy for selecting high-growing, stress-resistant algal strains with high photosynthetic capacity was proposed based on biocompatible extracellular polymeric substances (EPS) probes with aggregation-induced emission (AIE) properties. Specifically, AIE active EPS probes were synthesized for in-situ long-term monitoring of the EPS productivity at different algal growth stages. By coupling the AIE-based fluorescent techniques, algal cells were classified into four diverse populations based on their chlorophyll and EPS signals. Mechanistic studies on the sorted algal cells revealed their remarkable stress resistance and high expression of cell division, biopolymer production and photosynthesis-related genes. The sorted and subcultured algal cells consistently exhibited relatively higher growth rates and photosynthetic capacities, resulting in an increased (1.2 to 1.8-fold) algal biomass production, chlorophyll, and lipids. This study can potentially open new strategies to boost microalgal-based biofuel production.

Reactive oxygen species mediated extracellular polymeric substances production assisting the recovery of Thalassiosira pseudonana from polystyrene micro and nanoplastics exposure

As emerging pollutants in the aquatic environments, micro- and nano-plastics (MNPs) aroused widespread environmental concerns for their potential threats to the ecological health. Previous research has proved that microalgae growth could recover from the MNPs toxicities, in which the extracellular polymeric substances (EPS) might play the key role. In order to comprehensively investigate the recovery process of microalgae from MNPs stress and the effecting mechanisms of EPS therein, this study conducted a series of experiments by employing two sizes (0.1 and 1 μm) of polystyrene (PS) MNPs and the marine model diatom Thalassiosira pseudonana during 14 days. The results indicated: the pigments accumulations and photosynthetic recovery of T. pseudonana under MPs exposure showed in the early stage (4–5 days), while the elevation of reactive oxygen species (ROS) and EPS contents lasted longer time period (7–8 days). EPS was aggregated with MNPs particles and microalgal cells, corresponding to the increased settlement rates. More increase of soluble (SL)-EPS contents was found than bound (B)-EPS under MNPs exposure, in which the increase of the protein proportion and humic acid-like substances in SL-EPS was found, thus facilitating aggregates formation. ROS was the signaling molecule mediating the overproduction of EPS. The transcriptional results further proved the enhanced EPS biosynthesis on the molecular level. Therefore, this study elucidated the recovery pattern of microalgae from MNPs stress and linked “ROS-EPS production changes-aggregation formation” together during the growth recovery process, with important scientific and environmental significance.

Beyond growth: The significance of non-growth anabolism for microbial carbon-use efficiency in the light of soil carbon stabilisation

Microbial carbon-use efficiency (CUE) in soils captures carbon (C) partitioning between anabolic biosynthesis of microbial metabolites and catabolic C emissions (i.e. respiratory C waste). The use of C for biosynthesis provides a potential for the accumulation of microbial metabolic residues in soil. Recognised as a crucial control in C cycling, microbial CUE is implemented in the majority of soil C models. Due to the models' high sensitivity to CUE, reliable soil C projections demand accurate CUE quantifications. Current measurements of CUE neglect microbial non-growth metabolites, such as extracellular polymeric substances (EPS) or exoenzymes, although they remain in soil and could be quantitatively important. Here, we highlight that disregarding non-growth anabolism can lead to severe underestimations of CUE. Based on two case studies, we demonstrate that neglecting exoenzyme and EPS production underestimates CUE by more than 100% and up to 30%, respectively. By incorporating these case-specific values in model simulations, we observed that the model projects up to 34% larger SOC stocks over a period of 64 years when non-growth metabolites are considered for estimating CUE, highlighting the crucial importance of accurate CUE quantification. Our considerations outlined here challenge the current ways how CUE is measured and we suggest improvements concerning the quantification of non-growth metabolites. Research efforts should focus on (i) advancing CUE estimations by capturing the multitude of microbial C uses, (ii) improving techniques to quantify non-growth metabolic products in soil, and (iii) providing an understanding of dynamic metabolic C uses under different environmental conditions and over time. In the light of current discussion on soil C stabilisation mechanisms, we call for efforts to open the ‘black box’ of microbial physiology in soil and to incorporate all quantitative important C uses in CUE measurements.

Effect of Matricaria aurea Essential Oils on Biofilm Development, Virulence Factors and Quorum Sensing-Dependent Genes of Pseudomonas aeruginosa.

The emergence of drug-resistant microorganisms presents a substantial global public health threat. The increase in pathogens resistant to commonly prescribed antibiotics underscores the urgent requirement to explore alternative treatment strategies. This study adopts a novel approach by harnessing natural resources, specifically essential oils (EO), to combat bacterial pathogenicity. The primary aim of this research was to analyze the chemical composition of the aerial part of the Matricaria aurea (M. aureas) EO and evaluate its potential for inhibiting quorum sensing (QS) and disrupting biofilm formation in Pseudomonas aeruginosa (P. aeruginosa). The gas chromatography-mass spectrometry (GCMS) analysis unveiled that α-bisabolol oxide A constituted the predominant portion, comprising 64.8% of the total, with β-bisabolene at 6.3% and α-farnesene at 4.8% following closely behind. The antibiofilm efficacy was observed at concentrations of 0.3, 0.15, and 0.08 mg/mL, demonstrating negligible effects on cell viability. Furthermore, the EO from M. aurea effectively inhibited the formation of P. aeruginosa biofilms by diminishing aggregation, hydrophobicity, and swarming motility. Significantly, the EO treatment resulted in a conspicuous decrease in the production of pyocyanin, rhamnolipid, and extracellular polymeric substances (EPS), along with a reduction in the enzymatic activity of protease and chitinase. The EO effectively hindered QS by disrupting QS mechanisms, resulting in a marked decline in the secretion of N-Acyl homoserine lactone (AHL) molecules and the expression of phazA1 and aprA genes. This investigation offers compelling evidence supporting the potential of M. aurea EO as a promising therapeutic candidate for addressing infectious diseases induced by biofilm formation.

The potential effects of N-Acyl homoserine lactones on aerobic sludge granulation during phenolic wastewater treatment

The formation of aerobic granular sludge (AGS) is relatively difficult during the treatment of refractory wastewater, which generally shows small granular sizes and poor stability. The formation of AGS is regulated by N-Acyl homoserine lactones (AHLs)-mediated quorum sensing (QS). However, the potential role of AHLs in AGS formation under the toxic stress of refractory pollutants and the heterogeneity in the distribution and function of AHLs across different aggregates are not well understood. This study investigated the potential effects of AHLs on the formation of AGS during phenolic wastewater treatment. The distribution and succession of AHLs across varying granular sizes and development stages of AGS were investigated. Results showed that AGS was successfully formed in 13 days with an average granular size of 335 ± 39 μm and phenol removal efficiency of >99%. The levels of AHLs initially increased and then decreased. C4-HSL and 3-oxo-C10-HSL were enriched in large granules, suggesting they may play a pivotal role in regulating the concentration and composition of extracellular polymeric substances (EPS). The content of EPS constantly increased to 149.4 mg/gVSS, and protein (PN) was enriched in small and large granules. Luteococcus was the dominant genus constituting up to 62% after the granulation process, and exhibited a strong association with C4-HSL. AHLs might also regulate the bacterial community responsible for EPS production, and pollutant removal, and facilitate the proliferation of slow-growing microorganisms, thereby enhancing the formation of AGS. The synthesis and dynamics of AHLs were mainly governed by AHLs-producing bacterial strains of Rhodobacter and Pseudomonas, and AHLs-quenching strains of Flavobacterium and Comamonas. C4-HSL and 3-oxo-C10-HSL might be the major contributors to promoting sludge granulation under phenol stress and play critical roles in large granules. These findings enhance our understanding of the roles that AHLs play in sludge granulation under toxic conditions.