Linoleic acid inhibits Pseudomonas aeruginosa biofilm formation by activating diffusible signal factor-mediated quorum sensing.

Bacterial biofilm formation causes serious problems in various fields of medical, clinical, and industrial settings. Antibiotics and biocide treatments are typical methods used to remove bacterial biofilms, but biofilms are difficult to remove effectively from surfaces due to their increased resistance. An alternative approach to treatment with antimicrobial agents is using biofilm inhibitors that regulate biofilm development without inhibiting bacterial growth. In the present study, we found that linoleic acid (LA), a plant unsaturated fatty acid, inhibits biofilm formation under static and continuous conditions without inhibiting the growth of Pseudomonas aeruginosa. LA also influenced the bacterial motility, extracellular polymeric substance production, and biofilm dispersion by decreasing the intracellular cyclic diguanylate concentration through increased phosphodiesterase activity. Furthermore, quantitative gene expression analysis demonstrated that LA induced the expression of genes associated with diffusible signaling factor-mediated quorum sensing that can inhibit or induce the dispersion of P. aeruginosa biofilms. These results suggest that LA is functionally and structurally similar to a P. aeruginosa diffusible signaling factor (cis-2-decenoic acid) and, in turn, act as an agonist molecule in biofilm dispersion.

Interlayered Forward Osmosis Membranes with Ti3C2Tx MXene and Carbon Nanotubes for Enhanced Municipal Wastewater Concentration.

Forward osmosis (FO) hybrid systems have the potential to simultaneously recover nutrients and water from wastewater. However, the lack of membranes with high permeability and selectivity has limited the development and scale-up of these hybrid systems. In this study, we fabricated a novel thin-film nanocomposite membrane featuring an interlayer of Ti3C2Tx MXene intercalated with carbon nanotubes (M/C-TFNi). Owing to the enhanced confinement effect on interfacial degassing and increased amine monomer sorption by the interlayer, the resulting M/C-TFNi FO membrane has a greater degree of cross-linking and roughness. In comparison with the thin-film composite (TFC) membrane without an interlayered structure, the M/C-TFNi membrane attained a water flux that was four times higher and a lower specific salt flux. Notably, the M/C-TFNi membrane exhibited excellent concentration efficiency for real municipal wastewater and enhanced rejection of ammonia nitrogen, which breaks the permeability-selectivity upper bound. This study provides a new avenue for the rational design and development of high-performance FO membranes for environmental applications.

A Critical Review on Thin-Film Nanocomposite Membranes with Interlayered Structure: Mechanisms, Recent Developments, and Environmental Applications.

The separation properties of polyamide reverse osmosis and nanofiltration membranes, widely applied for desalination and water reuse, are constrained by the permeability-selectivity upper bound. Although thin-film nanocomposite (TFN) membranes incorporating nanomaterials exhibit enhanced water permeance, their rejection is only moderately improved or even impaired due to agglomeration of nanomaterials and formation of defects. A novel type of TFN membranes featuring an interlayer of nanomaterials (TFNi) has emerged in recent years. These novel TFNi membranes show extraordinary improvement in water flux (e.g., up to an order of magnitude enhancement) along with better selectivity. Such enhancements can be achieved by a wide selection of nanomaterials, ranging from nanoparticles, one-/two-dimensional materials, to interfacial coatings. The use of nanostructured interlayers not only improves the formation of polyamide rejection layers but also provides an optimized water transport path, which enables TFNi membranes to potentially overcome the longstanding trade-off between membrane permeability and selectivity. Furthermore, TFNi membranes can potentially enhance the removal of heavy metals and micropollutants, which is critical for many environmental applications. This review critically examines the recent developments of TFNi membranes and discusses the underlying mechanisms and design criteria. Their potential environmental applications are also highlighted.

Implications of Chemical Reduction Using Hydriodic Acid on the Antimicrobial Properties of Graphene Oxide and Reduced Graphene Oxide Membranes.

The antimicrobial properties of graphene-based membranes such as single-layer graphene oxide (GO) and modified graphene oxide (rGO) on top of cellulose ester membrane are reported in this study. rGO membranes are made from GO by hydriodic acid (HI) vapor treatment. The antibacterial properties are tested after 3 h contact time with selected model bacteria. Complete bacterial cell inactivation is found only after contact with rGO membranes, while no significant bacterial inactivation is found for the control i) GO membrane, ii) the mixed cellulose ester support, and the iii) rGO membrane after additional washing that removes the remaining HI. This indicates that the antimicrobial effect is neither caused by the graphene nor the membrane support. The antimicrobial effect is found to be conclusively linked to the HI eliminating microbial growth, at concentrations from 0.005%. These findings emphasize the importance of caution in the reporting of antimicrobial properties of graphene-based surfaces.

Antibacterial application of covalently immobilized photosensitizers on a surface.

Singlet oxygen produced by irradiating photosensitizers (PSs) can be used to kill pathogens during water treatment. Chemical immobilization of the PSs on surfaces can maintain their disinfection function long-term. In this study, two model PSs (rose bengal (RB) and hematoporphyrin (HP)) were immobilized on a glass surface using a silane coupling agent with an epoxide group, and their antibacterial properties were analyzed. Fourier transform infrared spectroscopy demonstrated that a covalent bond formed between the epoxide group and hydroxyl group in the PSs. A large proportion of the immobilized PSs (approximately 50%) was active in singlet oxygen production, which was evidenced by a comparative analysis with free PSs. RB was more effective at producing singlet oxygen than HP. The immobilized PSs were durable in terms of repeated use. On the other hand, singlet oxygen produced by the PSs was effective at killing bacteria, mostly for Gram-positive bacteria (> 90% death for 2 h of irradiation), by damaging the cell membrane. The preferable antibacterial property against Gram-positive bacteria compared with that against Gram-negative bacteria suggested efficient penetrability of singlet oxygen across the cell membrane, which led to cell death. Taken together, it was concluded that immobilization of PSs on surfaces using the silane coupling agent proposed in this study was effective at killing Gram-positive bacteria by forming singlet oxygen.

Identifying the Nonradical Mechanism in the Peroxymonosulfate Activation Process: Singlet Oxygenation Versus Mediated Electron Transfer.

Select persulfate activation processes were demonstrated to initiate oxidation not reliant on sulfate radicals, although the underlying mechanism has yet to be identified. This study explored singlet oxygenation and mediated electron transfer as plausible nonradical mechanisms for organic degradation by carbon nanotube (CNT)-activated peroxymonosulfate (PMS). The degradation of furfuryl alcohol (FFA) as a singlet oxygen (1O2) indicator and the kinetic retardation of FFA oxidation in the presence of l-histidine and azide as 1O2 quenchers apparently supported a role of 1O2 in the CNT/PMS system. However, the 1O2 scavenging effect was ascribed to a rapid PMS depletion by l-histidine and azide. A comparison of CNT/PMS and photoexcited Rose Bengal (RB) excluded the possibility of singlet oxygenation during heterogeneous persulfate activation. In contrast to the case of excited RB, solvent exchange (H2O to D2O) did not enhance FFA degradation by CNT/PMS and the pH- and substrate-dependent reactivity of CNT/PMS did not reflect the selective nature of 1O2. Alternatively, concomitant PMS reduction and trichlorophenol oxidation were achieved when PMS and trichlorophenol were physically separated in two chambers using a conductive vertically aligned CNT membrane. This result suggested that CNT-mediated electron transfer from organics to persulfate was primarily responsible for the nonradical degradative route.

Expression of the nirS, hzsA, and hdh Genes in Response to Nitrite Shock and Recovery in Candidatus Kuenenia stuttgartiensis.

In this study, Candidatus Kuenenia stuttgartiensis were subjected to distinct nitrite shocks (66 (control), 200, 300, 400, and 500 mg N/L), and the responses of mRNA levels of cytochrome cd1 nitrite/nitric oxide oxidoreductase (nirS), hydrazine synthase (hzsA), and hydrazine dehydrogenase (hdh) were assessed. Changes in the hydrazine dehydrogenase (HDH) protein level were monitored. At 200 mg NO2(-)-N/L, the normalized specific anaerobic ammonium-oxidizing activity (nSAA) slightly increased relative to the control despite a significant decrease in nirS, hzsA, and hdh mRNA levels. When nitrite increased to 300 and 400 mg N/L, increased nirS, hzsA, and hdh mRNA levels were observed, but the nSAA decreased, relative to the 200 mg NO2(-)-N/L exposure. HDH protein detection revealed that Candidatus Kuenenia stuttgartiensis attempted to yield high enzyme levels by stimulating mRNA synthesis to resist the nitrite-induced stress. On 500 mg NO2(-)-N/L shock, the nirS, hzsA, and hdh mRNA levels decreased, alongside decreased nSAA and HDH levels. Although the mRNA levels did not always coincide with activities, our findings advance understanding of the mechanisms that anammox bacteria use to cope with nitrite inhibition at the transcriptional and translational levels, which will improve the diagnostic accuracy of bioreactor failures when nitrite accumulation occurs.

Metagenomes reveal microbial structures, functional potentials, and biofouling-related genes in a membrane bioreactor.

Metagenomic sequencing was used to investigate the microbial structures, functional potentials, and biofouling-related genes in a membrane bioreactor (MBR). The results showed that the microbial community in the MBR was highly diverse. Notably, function analysis of the dominant genera indicated that common genes from different phylotypes were identified for important functional potentials with the observation of variation of abundances of genes in a certain taxon (e.g., Dechloromonas). Despite maintaining similar metabolic functional potentials with a parallel full-scale conventional activated sludge (CAS) system due to treating the identical wastewater, the MBR had more abundant nitrification-related bacteria and coding genes of ammonia monooxygenase, which could well explain its excellent ammonia removal in the low-temperature period. Furthermore, according to quantification of the genes involved in exopolysaccharide and extracellular polymeric substance (EPS) protein metabolism, the MBR did not show a much different potential in producing EPS compared to the CAS system, and bacteria from the membrane biofilm had lower abundances of genes associated with EPS biosynthesis and transport compared to the activated sludge in the MBR.

A survey of biofilms on wastewater aeration diffusers suggests bacterial community composition and function vary by substrate type and time.

Aeration diffusers in wastewater treatment plants generate air bubbles that promote mixing, distribution of dissolved oxygen, and microbial processing of dissolved and suspended matter in bulk solution. Biofouling of diffusers represents a significant problem to wastewater treatment plants because biofilms decrease oxygen transfer efficiency and increase backpressure on the blower. To better understand biofouling, we conducted a pilot study to survey the bacterial community composition and function of biofilms on different diffuser substrates and compare them to those in the bulk solution. DNA was extracted from the surface of ethylene-propylene-diene monomer (EPDM), polyurethane, and silicone diffusers operated for 15 months in a municipal treatment plant and sampled at 3 and 9 months. The bacterial community composition and function of the biofilms and bulk solution were determined by amplifying the 16S rRNA genes and pyrosequencing the amplicons and raw metagenomic DNA. The ordination plots and dendrograms of the 16S rRNA and functional genes showed that while the bacterial community composition and function of the bulk solution was independent of sampling time, the composition and function of the biofilms differed by diffuser type and testing time. For the EPDM and silicone diffusers, the biofilm communities were more similar in composition to the bulk solution at 3 months than 9 months. In contrast, the bacteria on the polyurethane diffusers were more dissimilar to the bulk solution at 3 months than 9 months. Taken together, the survey showed that the community composition and function of bacterial biofilms depend on the diffuser substrate and testing time, which warrants further elucidation.

Design, synthesis and biological evaluation of 4-(alkyloxy)-6-methyl-2H-pyran-2-one derivatives as quorum sensing inhibitors.

Novel pyrone-derived quorum sensing (QS) ligands to inhibit the binding of OdDHL to the LasR of Pseudomonas aeruginosa were designed, synthesized and evaluated. Among the analogs, the most potent compound 8 exhibited strong in vitro inhibitory activities against biofilm formation and down-regulated OdDHL/LasR-associated genes by 35-67%. The binding mode of 8 in silico was highly similar to that of the crystal ligand OdDHL in the active site of LasR.

Use of rhamnolipid biosurfactant for membrane biofouling prevention and cleaning.

Rhamnolipids were evaluated as biofouling reducing agents in this study. The permeability of the bacterial outer membrane was increased by rhamnolipids while the growth rate of Pseudomonas aeruginosa was not affected. The surface hydrophobicity was increased through the release of lipopolysaccharides and extracellular polymeric substances from the outer cell membrane. Rhamnolipids were evaluated as agents for the prevention and cleaning of biofilms. A high degree of biofilm detachment was observed when the rhamnolipids were used as a cleaning agent. In addition, effective biofilm reduction occurred when rhamnolipids were applied to various species of Gram-negative bacteria isolated from seawater samples. Biofilm reduction using rhamnolipids was comparable to commercially available surfactants. In addition, 20% of the water flux was increased after rhamnolipid treatment (300 µg ml(-1), 6 h exposure time) in a dead-end filtration system. Rhamnolipids appear to have promise as biological agents for reducing membrane biofouling.

Inhibition of biofouling by modification of forward osmosis membrane using quaternary ammonium cation.

In the operation of the forward osmosis (FO) process, biofouling of the membrane is a potentially serious problem. Development of an FO membrane with antibacterial properties could contribute to a reduction in biofouling. In this study, quaternary ammonium cation (QAC), a widely used biocidal material, was conjugated with a silane coupling agent (3-(trimethoxysilyl)-propyldimethyloctadecyl ammonium chloride) and used to modify an FO membrane to confer antibacterial properties. Fourier transform infrared spectroscopy (FT-IR) demonstrated that the conjugated QAC was successfully immobilized on the FO membrane via covalent bonding. Bacterial viability on the QAC-modified membrane was confirmed via colony count method and visualized via bacterial viability assay. The QAC membrane decreased the viability of Escherichia coli to 62% and Staphylococcus aureus to 77% versus the control membrane. Inhibition of biofilm formation on the QAC modified membrane was confirmed via anti-biofilm tests using the drip-flow reactor and FO unit, resulting in 64% and 68% inhibition in the QAC-modified membrane against the control membrane, respectively. The results demonstrate the effectiveness of the modified membrane in reducing bacterial viability and inhibiting biofilm formation, indicating the potential of QAC-modified membranes to decrease operation costs incurred by biofouling.

Comparison of bacterial communities of biofilms formed on different membrane surfaces.

Bacterial biofilm communities formed on different membrane surfaces were investigated based on 16S rRNA gene sequence analysis. The biofilm communities were distinct from those of mixed-liquor and consisted mainly of Beta- and Gammaproteobacteria. Sequences of Xathomonas and Aquabacterium were mostly retrieved from the biofilm samples rather than from the mixed liquor. Furthermore, statistical analyses demonstrated the importance of a physico-chemical property of membrane, surface roughness, in structuring the bacterial biofilm communities.

Isolation and characterization of novel 3,3'-iminodipropionitrile biodegrading Paracoccus communis, from an industrial wastewater treatment bioreactor.

Until now, bacteria able to degrade, 3,3'-iminodipropionitrile (IDPN), a neurotoxin that destroys vestibular hair cells, causing ototoxicity, culminating in irreversible movement disorders, had never been isolated. The aim of this study was to isolate a novel IDPN-biodegrading microorganism and characterize its metabolic pathway. Enrichment was performed by inoculating activated sludge from a wastewater treatment bioreactor that treated IDPN-contaminated wastewater in M9 salt medium, with IDPN as the sole carbon source. A bacterial strain with a spherical morphology that could grow at high concentrations was isolated on a solid medium. Growth of the isolated strain followed the Monod kinetic model. Based on the 16S rRNA gene, the isolate was Paracoccus communis. Whole-genome sequencing revealed that the isolated P. communis possessed the expected full metabolic pathway for IDPN biodegradation. Transcriptome analyses confirmed the overexpression of the gene encoding hydantoinase/oxoprolinase during the exponential growth phase under IDPN-fed conditions, suggesting that the enzyme involved in cleaving the imine bond of IDPN may promote IDPN biodegradation. Additionally, the newly discovered P. communis isolate seems to metabolize IDPN through cleavage of the imine bond in IDPN via nitrilase, nitrile hydratase, and amidase reactions. Overall, this study lays the foundation for the application of IDPN-metabolizing bacteria in the remediation of IDPN-contaminated environments.

General and rare bacterial taxa demonstrating different temporal dynamic patterns in an activated sludge bioreactor.

Temporal variation of general and rare bacterial taxa was investigated using pyrosequencing of 16S rRNA gene from activated sludge samples collected bimonthly for a two-year period. Most of operational taxonomic units (OTUs) were allocated to rare taxa (89.6%), but the rare taxa comprised a small portion of the community in terms of abundance of sequences analyzed (28.6%). Temporal variations in OTUs richness significantly differed between the two taxa groups in which the rare taxa showed a higher diversity and a more fluctuating pattern than the general taxa. Furthermore, the two taxa groups were constrained by different explanatory variables: influent BOD, effluent BOD, and DO were the significant (P < 0.05) parameters affecting the pattern of the general taxa, while temperature was the factor for the rare taxa. Over the test period, the general taxa persisted for a longer time (i.e., lower turnover rate) in the bioreactor than the rare taxa. In conclusion, this study demonstrated clear differences in temporal dynamic patterns for the general and rare bacterial taxa in an activated sludge bioreactor, which would be a foundation for better understanding the bacterial ecology of activated sludge.

Rapid enrichment of (homo)acetogenic consortia from animal feces using a high mass-transfer gas-lift reactor fed with syngas.

A gas-lift reactor having a high mass transfer coefficient (k(L)a = 80.28 h(-1)) for a relatively insoluble gas (carbon monoxide; CO) was used to enrich (homo)acetogens from animal feces. Samples of fecal matter from cow, rabbit, chicken, and goat were used as sources of inoculum for the enrichment of CO and H(2) utilizing microbial consortia. To confirm the successful enrichment, the Hungate roll tube technique was employed to count and then isolate putative CO utilizers. The results of this work showed that CO and H(2) utilizing consortia were established for each inoculum source after 8 days. The number of colony-forming units in cow, rabbit, chicken, and goat fecal samples were 3.83 × 10(9), 1.03 × 10(9), 8.3 × 10(8), and 3.25 × 10(8) cells/ml, respectively. Forty-two colonies from the animal fecal samples were screened for the ability to utilize CO/H(2). Ten of these 42 colonies were capable of utilizing CO/H(2). Five isolates from cow feces (samples 5, 6, 8, 16, and 22) were highly similar to previously unknown (homo)acetogen, while cow-7 has shown 99 % similarity with Acetobacterium sp. as acetogens. On the other hand, four isolates from chicken feces (samples 3, 8, 10, and 11) have also shown high CO/H(2) utilizing activity. Hence, it is expected that this research could be used as the basis for the rapid enrichment of (homo)acetogenic consortia from various environmental sources.

Mitigating from COVID-19 during Intercollegiate Sports Match.

The coronavirus disease 2019 pandemic has posed a significant threat not only to health outcomes but also to other societal sectors, including the educational system. Apart from youth education, colleges and universities are characterized by the integration of in-depth theoretical and practical knowledge in young adulthood. Our observations in this study suggest that college fairs, sports matches, and extracurricular activities can be safely resumed when population-level immunity has reached herd protection.

Rapid and sensitive detection of gram-negative bacteria using surface-immobilized polymyxin B.

Although detection of gram-negative bacteria (GNB) in body fluids is important for clinical purpose, traditional gram staining and other recently developed methods have inherent limitations in terms of accuracy, sensitivity, and convenience. To overcome the weakness, this study proposed a method detecting GNB based on specific binding of polymyxin B (PMB) to lipopolysaccharides (LPS) of GNB. Fluorescent microscopy demonstrated that surface immobilized PMB using a silane coupling agent was possible to detect fluorescent signal produced by a single Escherichia coli (a model GNB) cell. Furthermore, the signal was selective enough to differentiate between GNB and gram-positive bacteria. The proposed method could detect three cells per ml within one hour, indicating the method was very sensitive and the sensing was rapid. These results suggest that highly multifold PMB binding on each GNB cell occurred, as millions of LPS are present on cell wall of a GNB cell. Importantly, the principle used in this study was realized in a microfluidic chip for a sample containing E. coli cells suspended in porcine plasma, demonstrating its potential application to practical uses. In conclusion, the proposed method was accurate, sensitive, and convenient for detecting GNB, and could be applied clinically.

Biofilm characteristics for providing resilient denitrification in a hydrogen-based membrane biofilm reactor.

In a hydrogen-based membrane biofilm reactor (H2-MBfR), the biofilm thickness is considered to be one of the most important factors for denitrification. Thick biofilms in MBfRs are known for low removal fluxes owing to their resistance to substrate transport. In this study, the H2-MBfR was operated under various loading rates of oxyanions, such as NO3-N, SO4-S, and ClO4- at an H2 flux of 1.06 e- eq/m2-d. The experiment was initiated with NO3-N, SO4-S, and ClO4- loadings of 0.464, 0.026, and 0.211 e- eq/m2-d, respectively, at 20 °C. Under the most stressful conditions, the loading rates increased simultaneously to 1.911, 0.869, and 0.108 e- eq/m2-d, respectively, at 10 °C. We observed improved performance in significantly thicker biofilms (approximately 2.7 cm) compared to previous studies using a denitrifying H2-MBfR for 120 days. Shock oxyanion loadings led to a decrease in total nitrogen (TN) removal by 20 to 30%, but TN removal returned to 100% within a few days. Similarly, complete denitrification was observed, even at 10 °C. The protective function and microbial diversity of the thick biofilm may allow stable denitrification despite stress-imposing conditions. In the microbial community analysis, heterotrophs were dominant and acetogens accounted for 11% of the biofilm. Metagenomic results showed a high abundance of functional genes involved in organic carbon metabolism and homoacetogenesis. Owing to the presence of organic compounds produced by acetogens and autotrophs, heterotrophic denitrification may occur simultaneously with autotrophic denitrification. As a result, the total removal flux of oxyanions (1.84 e- eq/m2-d) far exceeded the H2 flux (1.06 e- eq/m2-d). Thus, the large accumulation of biofilms could contribute to good resilience and enhanced removal fluxes.

Inhibiting bacterial biofilm formation by stimulating c-di-GMP regulation using citrus peel extract from Jeju Island.

Biofilms consist of single or multiple species of bacteria embedded in extracellular polymeric substances (EPSs), which affect the increase in antibiotic resistance by restricting the transport of antibiotics to the bacterial cells. An alternative approach to treatment with antimicrobial agents is using biofilm inhibitors that regulate biofilm development without inhibiting bacterial growth. In this study, we found that citrus peel extract from Jeju Island (CPEJ) can inhibit bacterial biofilm formation. According to the results, CPEJ concentration-dependently reduces biofilm formation without affecting bacterial growth. Additionally, CPEJ decreased the production of extracellular polymeric substances but increased bacterial swarming motility. These results led to the hypothesis that CPEJ can reduce intracellular bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) concentration. The results showed that CPEJ significantly reduced the c-di-GMP level through increased phosphodiesterase activity. Altogether, these findings suggest that CPEJ as a biofilm inhibitor has new potential for pharmacological (e.g. drug and medication) and industrial applications (e.g. ship hulls, water pipes, and membrane processes biofouling control).