Extracellular Substances Research Articles

Mechanistic Insight into the Anti-Bacterial/Anti-Biofilm Effects of Low Chlorhexidine Concentrations on Enterococcus faecalis—In Vitro Study

Background: Endodontic treatment failures are often linked to the persistence of Enterococcus faecalis in the root canal system. This study aimed to investigate the antibacterial/antibiofilm mechanism of chlorhexidine (CHX), particularly at low concentrations, against E. faecalis, to improve endodontic treatment protocols. Methods: The antibacterial activity of CHX (0.125–20 μg/mL) was evaluated against E. faecalis ATCC 29212 using various assays, including planktonic growth inhibition, colony-forming units (CFUs), membrane permeability and potential assays, high-resolution scanning electron microscopy (HR-SEM), confocal laser scanning microscopy of biofilms, biomass and metabolic activity assays on matured biofilm, and quantitative real-time PCR for gene expression. Statistical analysis was performed using Student’s t-test and ANOVA. Results: CHX demonstrated concentration-dependent inhibition of E. faecalis, significantly reducing planktonic growth and CFUs. Membrane assays showed increased permeability and depolarization, indicating damage. HR-SEM revealed morphological changes, such as pore formation, while confocal microscopy showed a reduction in biofilm mass and extracellular substances. Gene expression analysis indicated the downregulation of virulence genes and upregulation of stress response genes. Conclusions: CHX at low concentrations disrupts E. faecalis at multiple levels, from membrane disruption to gene expression modulation, affecting mature biofilm. These findings support the refinement of endodontic disinfection protocols to reduce microbial persistence.

Adaptation of Chlorella vulgaris immobilization on rice straw with liquid manure to create a sustainable feedstock for biogas production and potential feed applications

Rice straw (RS) is a widely available agricultural residue with significant potential for biogas production and feed applications; however, its poor digestibility and nutritional value limit its utilization. This study explores an innovative approach to enhance the digestibility and nutritional value of RS by cultivating Chlorella vulgaris through immobilization technology on RS, using liquid manure (LM) as an alternative to the traditional BG11 medium. The results showed an increase in chlorophyll a (Chl a) after 12 days for both the BG11 medium and LM-based treatments, from 0.13 to 0.34 and 0.24 mg Chl a/g product (DM), respectively. Additionally, the immobilized microalgal biomass increased to 284.18 and 170.14 mg algal biomass/g product (DM), respectively. Soaking under microaerobic conditions during cultivation led to the partial degradation of RS. This, combined with the formed microalgal biofilm, contributed to an improved digestibility of the dry matter, reaching 69.1% and 65.9% for the final products based on the BG11 medium and LM mediums, respectively, compared to 52.1% for the raw RS. Furthermore, the crude protein and lipids contents were significantly improved with the potential for applications in feed, reaching 21.4% and 4.1% for the BG11 medium-based product, while they were observed to be 12.8% and 3.0%, respectively, for the LM-based product. Additionally, carbon-to-nitrogen ratio was significantly reduced compared to the raw RS. The higher digestibility and improved nutritional value contributed to increased biogas production, reaching 129.3 and 118.7 mL/g (TS) for the products based on the traditional medium and LM medium, respectively, compared to 86.7 mL/g (TS) for the raw RS. The immobilization mechanism and biofilm development could be attributed to the roughness of the RS and extracellular polymer substances. This study demonstrates that integrating C. vulgaris cultivation on RS with LM as a nutrient source not only enhances the digestibility and nutritional value of RS but also offers a sustainable waste management solution with potential applications in biogas production and animal feed.

Effect of low-frequency alternating magnetic field on exopolysaccharide production and antioxidant capacity of Pleurotus citrinopileatus by submerged fermentation.

The objective of this study was to investigate the effect of low-frequency alternating magnetic field (LF-AMF) on the production of extracellular polysaccharide (EPS) by submerged fermentation of Pleurotus citrinopileatus. The fermentation conditions optimized by the central composite design method were as follows: fermentation time of 6.18days, temperature of 28.28°C, shaking speed of 149.04 r/min, and inoculum amount of 8.43%. Under these conditions, a LF-AMF was applied to the submerged fermentation of P. citrinopileatus. When the intensity of LF-AMF was 40 Gs, the initial intervention time was 24h after inoculation, and the treatment time was 6h at one time, the mycelial biomass of P. citrinopileatus increased by 11.30%, and the EPS yield increased by 23.09% compared with the fermentation without LF-AMF treatment. The morphology of mycelium after LF-AMF treatment was observed by scanning electron microscopy. It was found that the surface of mycelium was wrinkled, and the structure of mycelium was loose, which might be more conducive to the production of EPS. Mycelium diameter decreased, and ATPase activity increased, indicating that LF-AMF had a positive effect on the production of EPS by P. citrinopileatus fermentation. Moreover, LF-AMF could improve the permeability of the mycelial cell membrane, facilitate the exchange of intracellular and extracellular substances, and increase the metabolic capacity of P. citrinopileatus. In vitro antioxidant test of EPS showed that LF-AMF treatment also improved its antioxidant capacity.

Surface-Adhered Microbubbles Enhance the Resistance of ANAMMOX Granular Sludge to Sulfamethoxazole Stress.

The granule-based anammox process has been reported to be more resistant to the stress of antibiotics; however, the underlying resistance mechanism is still not fully understood. In this study, we found that more microbubbles stably adhered to the surface layer of anammox granular sludge (AnGS, Gs) operating under long-term sulfamethoxazole (SMZ) stress of 2 mg/L, compared to that in the control reactor (Gc). The difference in covering content can be up to over three times (1.0 ± 0.1% vs 0.3 ± 0.0%). Batch tests showed that the coverage ratio of microbubbles on Gs reached approximately 32.5%, which significantly reduced SMZ transfer into AnGS due to the adsorption of SMZ by bubbles, thus alleviating the inhibition of anammox bacterial activity by 36.5%. The adhesion force between the microbubbles and the surface layer of Gs was found to be largely enhanced by 75.0% compared to that of Gc. The increased hydrophobicity of surface layer due to the increased extracellular polymer substance (EPS, mainly proteins) content, and the larger capillary force of surface layer, owing to the unique micronano structure, were identified as key factors responsible for the stable adhesion of microbubbles on the Gs. Consequently, this study demonstrated the vital roles of the surface-adhered microbubbles in resisting the stress of SMZ and shed light on the regulation and development of robust granule-based anammox processes.

Self-aggregation effect of the ternary system “Alga EPS-DOM-HMs” and the characterization of the self-adaptation metabolic response of microalgae

Heavy metals (HMs) present in the natural aquatic environment can form a ternary aggregate of “EPS-DOM-HMs” with the prevalent microalgae extracellular polymers substances (EPS) and macromolecular dissolved organic matters (DOMs), which show special molecular structure and biological interaction. This study reveals the formation of “EPS-TA-HMs” and the mechanism of their physiological and metabolic effects on Raphidocelis subcapitata. Results indicate that TA-Cr(III) can bind to EPS to form ternary aggregates with substances coexisting large and small hydrodynamic diameters and that the interactions are dominated by hydrophobic interactions of the protein binding to the pyrrole ring of the polyphenol and hydrogen bonding interactions formed by OC–(N R O). The protein structure of EPS has the largest proportion of proline, glycine, aspartic acid, and tryptophan. These interactions promoted the secretion of EPS components and reduced the growth inhibition of Raphidocelis subcapitata by 45.9 % compared with Cr(III) exposure. TEM analysis combined with EDS analysis indicated that Cr(III) was taken intracellularly and TA-Cr(III) was not. In addition, metabolomics analyses revealed that microalgae initiate adaptive mechanisms via the activation of a two-component system (i.e., maintenance of high metabolic activity). This study underscored the morphology of HMs in real aquatic environments and the mechanisms of metabolic effects on aquatic organisms.

The Predatory Properties of Bradymonabacteria, the Representative of Facultative Prey-Dependent Predators

Bradymonabacteria, as the representative of the facultative prey-dependent predators, were re-classified from the preceding Deltaproteobacteria into the phylum Myxococcota and proposed as a novel class named Bradymonadia. However, it was ambiguous whether their predatory pattern and properties were similar to those of the other myxobacterial predators. Therefore, the physiologic features were compared to determine the similarities and differences during the process of group attack and kin discrimination. Comparative genomic analyses were performed to conclude the core genome encoded commonly by bradymonabacteria, Myxococcia, and Polyangia. In conclusion, we proposed that bradymonabacteria have a predation pattern similar to the that of the representative of opportunistic predators like Myxococcus xanthus but with some subtle differences. Their predation was predicted to be initiated by the needle-less T3SS*, and the S-motility mediated by T4P also participated in the process. Meanwhile, their group attacks relied on cell contact and cell destiny. Inter-species (strains) kin discriminations occurred without the existence of T6SS. However, no extracellular lethal substance was detected in the fermentation liquor culture of bradymonabacteria, and the death of prey cells could only be observed when touched by their cells. Moreover, the prey-selective predation was observed when the predator encountered certain prey from Bacillus (G+), Algoriphagus (G−), and Nocardioides (G+). Bradymonabacteria can be regarded as a potential consumer and decomposer, and preying on many sea-dwelling or human pathogenic bacteria allows this group a broad application prospect in marine culture and clinical disease control. Our study will provide more evidence for its exploitations and applications.

Influence of solid residence time on the phosphorus adsorption of extracellular polymer substances

Influence of solid residence time (SRT) on phosphorus (P) adsorption of extracellular polymer substances (EPS) was investigated from the perspectives of the contents, compositions and properties of loosely bound and tightly bound extracellular polymers (i.e., LB-EPS and TB-EPS) and microbial community structure. The TP contents of EPS were 59.6±6.0 - 87.4±4.4 mg·g-1 VSS and 63.7% - 68.1% of those of sludge, which was still underestimated due to the residual TB-EPS protecting cell membrane integrity. The influences of SRT on the yield and compositions of TB-EPS were obviously greater than those of LB-EPS. Compared with the SRTs of 12 d and 18 d, the significant P storage enhancement of biological P removal (BPR) sludge with the SRT of 25 d was related with the increases of TB-EPS yield, polyP content and average chain length in TB-EPS. The P adsorption property of TB-EPS was slightly increased through increasing SRT, while the K, Mg and Ca adsorption properties were markedly reduced. Increasing extracellular P content could kept good BPR efficiency at longer SRTs, but the loss risk of phosphorus accumulating organism dominance was increased. Additionally, adding Mg and Ca salts might improve extracellular P adsorption at longer SRTs.

3D-printed controllable bio-accelerators with sustained release property to boost chromium (VI) inhibited denitrification recovery

Although soluble bio-accelerators have proven effective in mitigating Cr(VI) inhibition within denitrification system, issues persist in immobilizing bio-accelerators and making them slow-release for sustained regulation. In this study, a novel strategy was proposed to fabricate immobilized bio-accelerators with controlled structure, sustained release property by 3D printing technology. Notably, the sustained release of bio-accelerators from 3D-printed bio-accelerators (3DP-B) lasted for at least 144 h. Compared to control group, 3DP-B with basic components (3DP-BB) shortened the recovery time by 1.4 folds, and the COD and NO3--N removal efficiency was 36.5 % and 38.0 % higher than that of natural recovery. Correspondingly, the activity of key enzymes (nitrate reductase, nitrite reductase, nitric oxide reductase, and nitrous oxide reductase), electron transfer system activity and extracellular polymer substances of denitrification biofilm maintained at relatively high levels. Furthermore, introducing 60 mg·L-1 anthraquinone-2,6-disulfonate (AQDS) into the ink showed noticeable superiority on the bio-inhibition release over 1000 mg·L-1 AQDS. The released AQDS facilitated the electron transport capacity by 1.25 times compared with control group. The groundbreaking findings of this study could advance the development of 3D printing technology and utilization of bio-accelerators in the field of wastewater treatment.

Reverse Solute Diffusion Enhances Sludge Dewatering in Dead-End Forward Osmosis.

Wastewater treatment plants produce high quantities of excess sludge. However, traditional sludge dewatering technology has high energy consumption and occupies a large area. Dead-end forward osmosis (DEFO) is an efficient and energy-saving deep dewatering technology for sludge. In this study, the reverse osmosis of salt ions in the draw solution was used to change the sludge cake structure and further reduce its moisture content in cake by releasing the bound water in cell. Three salts, NaCl, KCl, and CaCl2, were added to the excess sludge feed solution to explore the roles of the reverse osmosis of draw solutes in DEFO. When the added quantities of NaCl and CaCl2 were 15 and 10 mM, respectively, the moisture content of the sludge after dewatering decreased from 98.1% to 79.7% and 67.3%, respectively. However, KCl did not improve the sludge dewatering performance because of the "high K and low Na" phenomenon in biological cells. The water flux increased significantly for the binary draw solute involving NaCl and CaCl2 compared to the single draw solute. The extracellular polymer substances in the sludge changed the structure of the filter cake to improve the formation of water channels and decrease osmosis resistance, resulting in an increase in sludge dewatering efficiency. These findings provide support for improving the sludge dewatering performance of DEFO.

Flagellar point mutation causes social aggregation in laboratory-adapted Bacillus subtilis under conditions that promote swimming.

Motility allows microbes to explore and maximize success in their environment; however, many laboratory bacterial strains have a reduced or altered capacity for motility. Swimming motility in Bacillus subtilis depends on peritrichous flagella and is carried out individually as cells move by biased random walks toward attractants. Previously, we adapted Bacillus subtilis strain 3610 to the laboratory for 300 generations in lysogeny broth (LB) batch culture and isolated lab-adapted strains. Strain SH2 is motility-defective and in broth culture forms large, frequently spherical aggregates of cells. A single point mutation in the flagellin gene hag that causes amino acid 259 to switch from A to T is necessary and sufficient to cause these social cell aggregates, and aggregation occurs between flagellated cells bearing this point mutation regardless of the strain background. Cells associate when bearing this mutation, but flagellar rotation is needed to pull associating cells into spherical aggregates. Using electron microscopy, we are able to show that the SH2 flagellar filament has limited polymorphism when compared to other flagellar structures. This limited polymorphism hinders the flagellum's ability to function as a motility apparatus but appears to alter its function to that of cell aggregation/adhesion. We speculate that the genotype-specific aggregation of cells producing HagA259T flagella could have increased representation in a batch-culture experiment by allowing similar cells to go through a transfer together and also that this mutation could serve as an early step to evolve sociality in the natural world.IMPORTANCEThe first life forms on this planet were prokaryotic, and the earliest environments were aquatic, and from these relatively simple starting conditions, complex communities of microbes and ultimately multicellular organisms were able to evolve. Usually, motile cells in aqueous environments swim as individuals but become social by giving up motility and secreting extracellular substances to become a biofilm. Here, we identify a single point mutation in the flagellum that is sufficient to allow cells containing this mutation to specifically form large, suspended groups of cells. The specific change in the flagellar filament protein subunits causes a unique change in the flagellar structure. This could represent a distinct way for closely related cells to associate as an early precursor to sociality.

The effect of extracellular polymeric substances on the distribution and transmission of antibiotic resistance genes treating antibiotic wastewater via microbial electrolysis cells

Microbial electrolysis cells (MEC) have emerged as a prominent technology for the treatment of antibiotics-containing wastewater in recent years. However, there remains a dearth of comprehensive exploration regarding the influence of extracellular polymers substances (EPS) on the distribution and transmission of antibiotic resistance genes (ARGs) in MEC. In this study, we quantified the distribution of ARGs in MEC by Fluorescence quantitative polymerase chain reaction and explored with emphasis on impact of EPS component on ARGs transmission at under different concentrations of roxithromycin. Results showed that the absolute abundance of ARGs in the electrode biofilm was 1–2 orders of magnitude higher than that in the anolyte. Specifically, EPS-associated ARGs accounted for 2.31%–11.18% of ARGs in electrode biofilm. The presence of elevated roxithromycin concentration led to electroactive microorganisms (Geobacter and Geothrix) as potential hosts of ARGs. In addition, both protein and polysaccharide content in the electrode biofilm increased with increasing roxithromycin concentration and showed positive correlations with EPS-associated ARGs. Fluorescence quenching experiments further elucidated that tryptophan and tyrosine residues in EPS could bind to ARGs effectively, contributing the hindering the ARGs transmission between hosts. Therefore, increased EPS content within electrode biofilm could reduce the concentration of ARGs present in anolyte while also influencing ARGs distribution throughout MEC. This study provides valuable insights into the distribution of ARGs in MEC systems and the role of EPS in regulating ARGs migration.

Catalyst-free regeneration of plasma-activated water via ultrasonic cavitation: Removing aggregation concealment of antibiotic-resistant bacteria with enhanced wastewater sustainability

Aggregation is a crucial factor in bacterial biofilm formation, and comprehending its properties is vital for managing waterborne antibiotic-resistant bacteria. In this study, we examined Methicillin-resistant Staphylococcus aureus (MRSA) cell aggregation under varying conditions and assessed the inactivation efficiency of a novel disinfection method, micro-nano bubbles plasma-activated water via ultrasonic stirring cavitation (MPAW-US), on aggregated MRSA cells. Aggregation efficiency increased over time and at low salt concentrations but diminished at higher concentrations. Elevated MRSA cell aggregation in actual water samples represented significant real-life biohazard risks. Unlike conventional disinfection, MPAW-US treatment exhibited minimal change in the inactivation rate constant despite protective outer layers. Enhanced inactivation efficiency results from the synergistic effects of increased intracellular oxidative stress damage and extracellular substance disruption, triggered by ultrasound-activated micro-nano bubbles that improve PAW reactivity and applicability. This approach neither induced MRSA cross-resistance to unfavorable conditions nor increased toxicity or regrowth potential of aggregative MRSA, utilizing ATP levels as potential regrowth capability indicators. Ultimately, this energy-efficient disinfection technology functions effectively across diverse temperature ranges, showcasing exceptional sterilization and nutritional bean sprout production after cyclic filtering, thereby promoting wastewater sustainability amidst carbon emission concerns.

A comparison study on membrane fouling in A/O-MBR and A/A-MBR at different mixed liquor-suspended solids concentrations

ABSTRACT Membrane fouling leads to decreased membrane flux, increases the frequency of membrane tissue replacement and membrane cleaning, and increases the operating cost of membrane bioreactor. In this study, the pollutant removal effects, membrane fouling differences and microbial characteristics of anaerobic/aerobic MBR (A/O-MBR) and anaerobic/anoxic MBR (A/A-MBR) were investigated at different mixed liquor suspended solids (MLSS) concentrations. The results showed that the chemical cleaning cycle of membrane contamination was 12, 28, 44 h and 24, 40, 104 h, respectively, and the cycle was prolonged with the increase of MLSS concentration (from 6000 to 9000 mg L−1). A/O-MBR was 1.4–2.4 times the rate of membrane fouling of A/A-MBR. In irreversible resistance, extracellular polymer substances (EPS) were the most significant contributors to membrane fouling. EPS concentration in A/A-MBR (118.33, 73.75, 54.26 mg/gMLSS) was lower than that in A/O-MBR (171.68, 91.92, 62.33 mg/gMLSS). Therefore, increasing MLSS concentration could mitigate membrane fouling. 16S rRNA high-throughput sequencing demonstrated that filamentous bacteria was the primary reason for the membrane fouling difference. Filamentous bacteria were more likely to be attached to the surface of the membrane, causing membrane fouling. The abundance percentage of filamentous bacteria in A/A-MBR was smaller than that in A/O-MBR. In summary, The excellent performance of A/A-MBR in membrane fouling behaviour, resistance analysis, EPS and microorganisms proved that A/A-MBR is more promising than A/O-MBR in wastewater nitrogen and phosphorus removal. This study can provide a theoretical basis for the application of MBR in the field of sewage treatment.

Toxic effects of the emerging Alexandrium pseudogonyaulax (Dinophyceae) on multiple trophic levels of the pelagic food web

The dinoflagellate Alexandrium pseudogonyaulax, a harmful algal bloom species, is currently appearing in increasing frequency and abundance across Northern European waters, displacing other Alexandrium species. This mixotrophic alga produces goniodomins (GDs) and bioactive extracellular substances (BECs) that may pose a threat to coastal ecosystems and other marine resources. This study demonstrated the adverse effects of A. pseudogonyaulax on four marine trophic levels, including microalgae (Rhodomonas salina), microzooplankton (Polykrikos kofoidii) and mesozooplankton (Acartia tonsa), as well as fish gill cells (RTgill-W1, Oncorhynchus mykiss), ultimately leading to enhanced mortality and cell lysis. Furthermore, cell-free supernatants collected from A. pseudogonyaulax cultures caused complete loss of metabolic activity in the RTgill-W1 cell line, indicating ichthyotoxic properties, while all tested GDs were much less toxic. In addition, cell-free supernatants of A. pseudogonyaulax led to cell lysis of R. salina, while all tested GDs were non-lytic. Finally, reduced egg hatching rates of A. tonsa eggs exposed to cell-free supernatants of A. pseudogonyaulax and impaired mobility of P. kofoidii and A. tonsa exposed to A. pseudogonyaulax were also observed. Altogether, bioassay results suggest that the toxicity of A. pseudogonyaulax is mainly driven by BECs and not by GDs, although further research into factors modulating the lytic activity of Alexandrium spp. are needed.

Correlative Effects on Nanoplastic Aggregation in Model Extracellular Biofilm Substances Investigated with Fluorescence Correlation Spectroscopy.

Recent studies show that biofilm substances in contact with nanoplastics play an important role in the aggregation and sedimentation of nanoplastics. Consequences of these processes are changes in biofilm formation and stability and changes in the transport and fate of pollutants in the environment. Having a deeper understanding of the nanoplastics-biofilm interaction would help to evaluate the risks posed by uncontrolled nanoplastic pollution. These interactions are impacted by environmental changes due to climate change, such as, e.g., the acidification of surface waters. We apply fluorescence correlation spectroscopy (FCS) to investigate the pH-dependent aggregation tendency of non-functionalized polystyrene (PS) nanoparticles (NPs) due to intermolecular forces with model extracellular biofilm substances. Our biofilm model consists of bovine serum albumin (BSA), which serves as a representative for globular proteins, and the polysaccharide alginate, which is a main component in many biofilms, in solutions containing Na+ with an ionic strength being realistic for fresh-water conditions. Biomolecule concentrations ranging from 0.5 g/L up to at maximum 21 g/L are considered. We use non-functionalized PS NPs as representative for mostly negatively charged nanoplastics. BSA promotes NP aggregation through adsorption onto the NPs and BSA-mediated bridging. In BSA-alginate mixtures, the alginate hampers this interaction, most likely due to alginate-BSA complex formation. In most BSA-alginate mixtures as in alginate alone, NP aggregation is predominantly driven by weaker, pH-independent depletion forces. The stabilizing effect of alginate is only weakened at high BSA contents, when the electrostatic BSA-BSA attraction is not sufficiently screened by the alginate. This study clearly shows that it is crucial to consider correlative effects between multiple biofilm components to better understand the NP aggregation in the presence of complex biofilm substances. Single-component biofilm model systems based on comparing the total organic carbon (TOC) content of the extracellular biofilm substances, as usually considered, would have led to a misjudgment of the stability towards aggregation.

Application of Nanomaterials and Related Drug Delivery Systems in Autophagy.

Autophagy, a lysosomal self-degradation pathway, plays a critical role in cellular homeostasis by degrading endogenous damaged organelles and protein aggregates into recyclable biological molecules. Additionally, it detoxifies extracellular toxic substances, including drugs and toxic materials, thereby preserving the stability of the intracellular environment. The swift progression of nanotechnology has led to an increased focus on understanding the relationship between nanomaterials and autophagy. The effects of various nanomaterials and nano drug delivery systems on autophagy and their biological functions have been preliminarily assessed, revealing that modulation of intracellular autophagy levels by these agents represents a novel cellular response mechanism. Notably, autophagy regulation based on nanomaterials or nano drug delivery systems for a range of diseases is currently the subject of extensive research. Given the close association between autophagy levels and tumors, the regulation of autophagy has emerged as a highly active area of research in the development of innovative tumor therapies. This review synthesizes the current understanding of the application of nanomaterials or nano drug delivery systems on autophagy and their potential biological functions, suggesting a new avenue for nanomaterial-based autophagy regulation.

Novel biostimulant bacterial exopolysaccharides production via solid-state fermentation as a valorisation strategy for agri-food waste.

Bacterial exopolysaccharides (EPS) are extracellular polymer-based substances recently defined as potential plant biostimulants, as they can increase nutrient uptake, water retention, and resistance to abiotic stress. As sugar-based substances, the bacteria producing them need to grow in a sugar-rich substrate. Hence, some agri-food by-products could be used as suitable carbon sources for EPS production as a cost-effective and more sustainable alternative to conventional substrates. Thus, this study aimed to produce EPS from specific bacterial strains through solid-state fermentation (SSF) using agri-food waste as a low-cost substrate. Six residues and five bacterial strains were tested in a lab-scale SSF system. From the assessed substrate-strain combinations, Burkholderia cepacia with ginger juice waste (GJW) resulted in the most promising considering several process parameters (EPS production, cumulative oxygen consumption, biomass growth, reducing sugars consumption). Also, dynamic monitoring of the system allowed for establishing 5days as a suitable fermentation time. Then, using response surface methodology (Box-Behnken design), the process was optimised based on airflow rate (AF), inoculum size (IS), and micronutrient concentration (MN). In this stage, the best conditions found were at 0.049 (± 0.014) L h-1 per gram of dry matter (DM) for AF, 8.4 (± 0.9) E + 09CFUg-1 DM for IS, and 0.07 (± 0.01) mL g-1 DM for MN, reaching up to 71.1 (± 3.2) mg crude EPS g-1 DM. Results show the potential of this approach to provide a new perspective on the value chain for the agri-food industry by introducing it to a circular economy framework.

Piedraia hortae: biofilm formation and its importance in the pathogenesis of Piedra nigra (black piedra)

BackgroundLittle is known about the ultrastructure of Piedraia hortae. ObjectiveTo examine a P. hortae colony with scanning electron microscopy and investigate possible contributions to its pathogenesis. ResultsOn low magnifications, two distinct aspects of the colony are identified, a compact area and a filamentous area. Analysis of the filamentous area demonstrates hyphae adhered by a thin reticular substance. A recurring finding is the adhesion between the fungal filaments in parallel. On high magnifications, the micro fibrillar substance adhering the hyphae to each other becomes very evident. Examination of the compact area shows the hyphae embedded in the reticular matrix forming a biofilm and the colony well adhered. On high magnification, it can be observed that the hyphae are within this fibrillar matrix, which has the same appearance as the filamentous substance that adheres the hyphae to each other. Study limitationsOnly one strain was examined. ConclusionsThe formation of biofilm with fungal structures and reticulated extracellular substance is important in the pathogenesis of Piedra nigra.

Insight into enhancing the performance of sludge dewatering using a novel flocculant CS–TA prepared through free radical-mediated conjugation

ABSTRACT Flocculation is one of the most significant conditioning methods for sludge dewatering. In the study, a novel flocculant CS–TA, prepared through free radical-mediated conjugation of tannic acid (TA) and chitosan (CS), was proposed to improve sludge dewatering. The characterisation using Fourier transform infra-red spectroscopy and X-ray diffraction analysis shows that the CS chain was the backbone of CS–TA, and the presence of CS–TA aromatic rings confirmed the conjugation of CS with TA. Moreover, the conditioning of CS–TA yielded the best dewatering performance at 30 mg g TS−1 with the water content of sludge cake by press filtration (Wsc) of 59.78% ± 0.3% and capillary suction time (CST) of 11.8s ± 0.35 s, compared to 98.2% ± 0.15% and 56.2 s ± 0.16 in raw sludge. The results of different influencing factors (e.g. pH and temperature) on flocculation efficiency indicated that CS–TA possessed the capacity for enhancing sludge dewaterability over a wide range of pH, and the optimal temperature was observed to be 35 °C. Furthermore, the increase of particle size and zeta potential implied the addition of CS–TA favoured the formation of larger particles charge neutralisation and adsorption bridging effect. In addition, extracellular polymer substances (EPS) analysis indicated that the decrease in the polysaccharide and protein contents in EPS after CS–TA addition could increase the relative hydrophobicity of sludge. Moreover, the contents of heavy metals in sludge and their leaching toxicity and environmental risk were reduced. This study provides comprehensive insights into the exploration of CS–TA for sludge dewatering and the maintenance of ecological security in an eco-friendly.

Simultaneous removal of nitrate nitrogen and ammonium nitrogen by partial denitrification coupled with anammox synergy by zero-valent iron

The implications of zero-valent iron (ZVI) on performance of partial denitrification (PD) coupled with anammox process (PD-anammox) and microbial community structures were assessed in present investigation. The results indicated that removal efficiencies of nitrate nitrogen, ammonium nitrogen and total nitrogen (TN) reached 86.28 ± 0.04%, 81.05 ± 0.06% and 82.90 ± 0.80%, respectively, with 30-day addition of ZVI under ratio of ZVI to nitrate (ZVI/N), carbon to nitrate (C/N) respectively for 3 and 2.3. Moreover, the content of extracellular polymer substance elevated by ZVI, and which could substitute carbon sources, mediated electron transfer and promoted nitrogen removal efficiency. Additionally, microbiological analysis revealed that ZVI enhanced nitrogen removal process by promoting the growth of genus Ignavibacterium and Denitratisoma, which coexist with anaerobic ammonia oxidation bacterium (AnAOB) and also induced other nitrogen removal process such as heterotrophic nitrate reduction to ammonium (DNRA), nitrate-dependent anaerobic ferrous oxide (NAFO), and iron-based anammox (Feammox) process. This study offers new insights into the enhancement of the nitrogen removal process by ZVI in PD-anammox process.