Synthesis Of Extracellular Polymeric Substances Research Articles

Role of Lactobacillus biofilms in Listeria monocytogenes adhesion to glass surfaces

Listeria monocytogenes can form long-lasting biofilms on food-contact surfaces. Lactic acid bacteria (LAB) have shown promise in antagonizing this microorganism in liquid media. However, the ecological relationships differ when cells are forming biofilms. In this work, we propose the use of Lactobacillus biofilms as surface “conditioners” to modulate the adhesion of L. monocytogenes. For this, the biofilm formation ability of Lactobacillus fermentum MP26 and Lactobacillus salivarius MP14 (human milk origin), fluorescently labeled by transfer of the mCherry-encoding pRCR12 plasmid, was first evaluated. Then, mature biofilms of these strains transformed with pRCR12 for expressing the fluorescent protein mCherry were used as adhesion substrate for GFP-tagged L. monocytogenes Scott A. The resulting biofilms were studied in terms of cellular population and attached biomass (cells plus matrix). Species distribution inside the biofilm structure was revealed by confocal laser scanning microscopy (CLSM). Although none of the Lactobacillus spp. strains reduced the adhesion of L. monocytogenes Scott A, species interactions seem to interfere with the synthesis of extracellular polymeric substances and species distribution inside the biofilms. In dual-species biofilms, CLSM images revealed that Lactobacillus cells were trapping those of L. monocytogenes Scott A. When surfaces were conditioned with Lactobacillus biofilms, the spatial distribution of L. monocytogenes Scott A cells was species-specific, suggesting these interactions are governing the ultimate biofilm structure. The results here obtained open new possibilities for controlling L. monocytogenes dispersal using these Lactobacillus spp. biofilms as a “natural” immobilization way. Whether species interactions could modify the virulence of L. monocytogenes still remains unclear.

The substrate-dependent regulatory effects of the AfeI/R system in Acidithiobacillus ferrooxidans reveals the novel regulation strategy of quorum sensing in acidophiles.

SummaryA LuxI/R‐like quorum sensing (QS) system (AfeI/R) has been reported in the acidophilic and chemoautotrophic Acidithiobacillus spp. However, the function of AfeI/R remains unclear because of the difficulties in the genetic manipulation of these bacteria. Here, we constructed different afeI mutants of the sulfur‐ and iron‐oxidizer A. ferrooxidans, identified the N‐acyl homoserine lactones (acyl‐HSLs) synthesized by AfeI, and determined the regulatory effects of AfeI/R on genes expression, extracellular polymeric substance synthesis, energy metabolism, cell growth and population density of A. ferrooxidans in different energy substrates. Acyl‐HSLs‐mediated distinct regulation strategies were employed to influence bacterial metabolism and cell growth of A. ferrooxidans cultivated in either sulfur or ferrous iron. Based on these findings, an energy‐substrate‐dependent regulation mode of AfeI/R in A. ferrooxidans was illuminated that AfeI/R could produce different types of acyl‐HSLs and employ specific acyl‐HSLs to regulate specific genes in response to different energy substrates. The discovery of the AfeI/R‐mediated substrate‐dependent regulatory mode expands our knowledge on the function of QS system in the chemoautotrophic sulfur‐ and ferrous iron‐oxidizing bacteria, and provides new insights in understanding energy metabolism modulation, population control, bacteria‐driven bioleaching process, and the coevolution between the acidophiles and their acidic habitats.

Synthesized effects of proteomic and extracellular polymeric substance (EPS) revealing the enhanced hydrogen production by formed biofilm of photo-fermentative bacteria

Photo-fermentative hydrogen production, the new energy production alternative, was greatly enhanced by formed biofilm. To understand the mechanism of enhancement, the intracellular proteome and extracellular polymeric substance (EPS)iiEPS: extracellular polymeric substanceiiPFHP: Photo-fermentative hydrogen productioniiiPFB: photo-fermentative bacteriaivQS: quorum sensingvCLSM: Confocal laser scanning microscopyviCR: Control reactorviiBR: Biofilm reactorviiiTEM: Transmission electron microscopeixXPS: X-ray photoelectron spectroscopyxFTIR: Fourier transform infrared spectroscopyxiSM: Supplementary materialsxiiDEPs: Differentially expressed proteinsxiiiTCA cycle: Tricarboxylic acid cyclexivGO: Gene ontologyxvKEGG: Kyoto encyclopedia of genes and genomes during biofilm formation were investigated in this work. Experimental results indicated that a possible and effective altered system could transfer light to hydrogen. Proteins were significantly regulated, for example those related with nitrogenase, flagellin, EPS transportation and DNA duplication were up-regulated while those concerned photosystem were down-regulated. It revealed these changes of proteins contributed to positive activity of key enzymes, improved communication system and increased total light utilization efficiency thus leading to enhanced capacity of hydrogen production. Besides above metabolic changes inside the cells, EPS secreted by the bacteria played an important role in hydrogen production and its yield decided the release of hydrogen. When EPS descended to a lower concentration during biofilm formation, it meant carbon source for EPS synthesis was reduced, and more energy and reducing power could be transferred into hydrogen energy. More importantly, this work found that composition and structure of EPS were efficiently influenced by the formation of biofilm, such as benzene and O-H structure, secondary protein structure and the kinds of protein, which were important to stable biofilm and efficient hydrogen production. Therefore, final hydrogen yield was improved by altered protein and EPS resulted from biofilm formation. This study demonstrated that formation of biofilm is an efficient, ecological and attracting way to the future bio-hydrogen production.

Links between S-adenosylmethionine and Agr-based quorum sensing for biofilm development in Listeria monocytogenes EGD-e.

Listeria monocytogenes is the causative agent of human listeriosis which has high hospitalization and mortality rates for individuals with weakened immune systems. The survival and dissemination of L. monocytogenes in adverse environments can be reinforced by the formation of biofilms. Therefore, this study aimed to understand the mechanisms underlying listerial biofilm development. Given that both nutrient availability and quorum sensing (QS) have been known as the factors influencing biofilm development, we hypothesized that the signal from a sentinel metabolite S‐adenosylmethionine (SAM) and Agr‐based QS could be synchronous in L. monocytogenes to modulate nutrient availability, the synthesis of extracellular polymeric substances (EPSs), and biofilm formation. We performed biofilm assays and quantitative real‐time PCR to investigate how biofilm volumes and the expression of genes for the synthesis of EPS were affected by SAM supplementation, agr deletion, or both. We found that exogenously applied SAM induced biofilm formation and that the expression of genes encoding the EPS synthesis machineries was regulated by SAM and/or Agr QS. Moreover, the gene transcription of components acting in the methyl cycle for SAM synthesis and Agr QS was affected by the signals from the other system. In summary, we reveal an interconnection at the transcriptional level between metabolism and QS in L. monocytogenes and highlight the critical role of metabolite‐oriented QS in biofilm development.

Antibiofilm activity of symbiotic Bacillus species associated with marine gastropods

PurposeGenerally, symbiotic marine bacteria are renowned for the synthesis of compounds with bioactive properties, and this has been documented in many previous studies. Therefore, the present study was aimed to isolate novel bacterial symbionts of gastropods that have the ability to synthesize bioactive compounds. These bioactive compounds could be used effectively as antibiofilm agents in order to overcome the problems associated with biofilm.MethodsThe bacteria associated with the surface of marine gastropods were isolated and characterized. Following this, the bacterial metabolites were extracted and their antibiofilm effect was evaluated on biofilm-forming bacteria on artificial substrates. Moreover, the biofilm-forming bacterium Alteromonas sp. was treated with the extracts of symbiotic bacteria in order to evaluate the influence of extracts over the synthesis of extracellular polymeric substance (EPS). Besides, the biologically active chemical constituents of the extracts were separated using thin-layer chromatography and subjected to gas chromatography and mass spectrometry (GC-MS) analysis for characterization.ResultsThree bacterial strains belonging to the species Bacillus firmus, Bacillus cereus and Bacillus subtilis were identified from the bacterial community associated with the gastropods. The antibiofilm assays revealed that the extract of three symbiotic bacteria significantly (p < 0.05) reduced the biofilm formation by the marine bacterium Alteromonas sp. on artificial materials. Also, the EPS synthesis by Alteromonas sp. was significantly inhibited due to symbiotic bacterial extract treatment. The chemical composition of the bioactive fraction isolated from the symbiotic bacteria extract revealed that most of the detected compounds were belonging to aromatic acid, fatty acid and carboxylic acid.ConclusionThe results of this study clearly revealed that the bacteria belonging to the above listed Bacillus species can be considered as a promising source of natural antibiofilm agents.

Novel Strategy for Improvement of the Bioleaching Efficiency of Acidithiobacillus ferrooxidans Based on the AfeI/R Quorum Sensing System

Acidithiobacillus ferrooxidans is an acidophilic and chemolithotrophic sulfur- and iron-oxidizing bacterium that has been widely used in the bioleaching process for extracting metals. Extracellular polymeric substances (EPS) are essential for bacteria-ore interactions, and the regulation of EPS synthesis could be an important way of influencing the efficiency of the bioleaching process. Therefore, exploring and utilizing the regulatory pathways of EPS synthesis to improve the bacterial bioleaching capability have posed a challenge in the study and application of bioleaching bacteria. Here, several engineering strains were constructed using genetic manipulation methods. And we revealed the regulatory function of the AfeI/R quorum sensing (QS) system in EPS synthesis and biofilm formation of A. ferrooxidans, and the AfeI/R-mediated EPS synthesis could influence bacteria-substrate interactions and the efficiency of bioleaching. Finally, an AfeI/R-mediated bioleaching model was proposed to illustrate the role of QS system in this process. This study provided new insights into and clues for developing highly efficient bioleaching bacteria and modulating the bioleaching process.

The antitoxin MqsA homologue in Pseudomonas fluorescens 2P24 has a rewired regulatory circuit through evolution.

The mqsRA operon encodes a toxin-antitoxin pair that was characterized to participate in biofilm and persister cell formation in Escherichia coli. Notably, the antitoxin MqsA possesses a C-terminal DNA-binding domain that recognizes the [5'-AACCT(N)2-4 AGGTT-3'] motif and acts as a transcriptional regulator controlling multiple genes including the general stress response regulator RpoS. However, it is unknown how the transcriptional circuits of MqsA homologues have changed in bacteria over evolutionary time. Here, we found mqsA in Pseudomonas fluorescens (PfmqsA) is acquired through horizontal gene transfer and binds to a slightly different motif [5'-TACCCT(N)3 AGGGTA-3'], which exists upstream of the PfmqsRA operon. Interestingly, an adjacent GntR-type transcriptional regulator, which was termed AgtR, is under negative control of PfMqsA. It was further demonstrated that PfMqsA reduces production of biofilm components through AgtR, which directly regulates the pga and fap operons involved in the synthesis of extracellular polymeric substances. Moreover, through quantitative proteomics analysis, we showed AgtR is a highly pleiotropic regulator that influences up to 252 genes related to diverse processes including chemotaxis, oxidative phosphorylation and carbon and nitrogen metabolism. Taken together, our findings suggest the rewired regulatory circuit of PfMqsA influences diverse physiological aspects of P. fluorescens 2P24 via the newly characterized AgtR.

Chemical Dispersant Enhances Microbial Exopolymer (EPS) Production and Formation of Marine Oil/Dispersant Snow in Surface Waters of the Subarctic Northeast Atlantic

A notable feature of the Deepwater Horizon oil spill was the unprecedented formation of marine oil snow (MOS) that was observed in large quantities floating on the sea surface and that subsequently sedimented to the seafloor. Whilst the physical and chemical processes involved in MOS formation remain unclear, some studies have shown that extracellular polymeric substances (EPS) play a role in this process. Here, we report that during exposure of subarctic northeast Atlantic seawater to a chemical dispersant, whether in the presence/absence of crude oil, the dispersant stimulates the production of significant quantities of EPS that we posit serves as a key building block in the formation of MOS. This response is likely conferred via de novo synthesis of EPS by natural communities of bacteria. We also describe the formation of marine dispersant snow (MDS) as a product of adding chemical dispersants to seawater. Differential staining confirmed that MDS, like MOS, is composed of glycoprotein, though MDS is more protein rich. Using barcoded-amplicon Illumina MiSeq sequencing, we analyzed, for the first time, the bacterial communities associated with MDS and report that their diversity is not significantly dissimilar to those associated with MOS aggregates. Our findings emphasize the need to conduct further work on the effects of dispersants when applied to oil spills at sea, particularly at different sites, and to determine how the product of this (i.e., MOS and MDS) affects the biodegradation of the oil.

Metagenomic quorum quenching enzymes affect biofilm formation of Candida albicans and Staphylococcus epidermidis.

Biofilm formation in the clinical environment is of increasing concern since a significant part of human infections is associated, and caused by biofilm establishment of (opportunistic) pathogens, for instance Candida albicans and Staphylococcus epidermidis. The rapidly increasing number of antibiotic-resistant biofilms urgently requires the development of novel and effective strategies to prevent biofilm formation ideally targeting a wide range of infectious microorganisms. Both, synthesis of extracellular polymeric substances and quorum sensing are crucial for biofilm formation, and thus potential attractive targets to combat undesirable biofilms.We evaluated the ability of numerous recently identified metagenome-derived bacterial quorum quenching (QQ) proteins to inhibit biofilm formation of C. albicans and S. epidermidis. Here, proteins QQ-5 and QQ-7 interfered with the morphogenesis of C. albicans by inhibiting the yeast-to-hyphae transition, ultimately leading to impaired biofilm formation. Moreover, QQ5 and QQ-7 inhibited biofilm formation of S. epidermidis; in case of QQ7 most likely due to induced expression of the icaR gene encoding the repressor for polysaccharide intercellular adhesin (PIA) synthesis, the main determinant for staphylococcal biofilm formation. Our results indicate that QQ-5 and QQ-7 are attractive potential anti-biofilm agents in the prevention and treatment of C. albicans and S. epidermidis mono-species biofilms, and potentially promising anti-biofilm drugs in also combating multi-species infections.

Effect of light intensity on the characteristics of algal-bacterial granular sludge and the role of N-acyl-homoserine lactone in the granulation

The effects of light intensity on the development of algal-bacterial granular sludge (ABGS) were investigated over a period of 12 weeks. The ABGS developed at low light intensity (142 ± 10 μmol m−2·s−1) exhibited excellent settling ability (SVI30 of 30.9 mL/g), COD and TN removal efficiencies (97.6% and 60.4%, respectively). High light intensity (316 ± 12 μmol m−2·s−1) accelerated granular biomass growth (5.3 g/L) and enhanced the TP removal efficiency (83.7%). Extracellular polymeric substance (EPS) analysis revealed that low light intensity induced more large weight distribution protein production (9–12 kDa and 50–150 kDa), predominantly tryptophan and aromatic proteins. Furthermore, N-acyl-homoserine lactones (AHLs) with a side chain ≤ C10 were commonly shared in the ABGS, and the ABGS developed at low light intensity had a higher C6- and 3OC8-HSL content, which effectively promoted the biofilm formation. The add-back studies showed that the AHLs facilitated the regulation of EPS synthesis. Statistical analysis indicated that the AHLs content had a close correlation with the EPS production, the 50th percentile of the particle size distribution and microbial community assembly, suggesting that AHLs-mediated quorum sensing have an important ecological role in EPS expression and algal-bacterial granulation. Overall, this study describes the ABGS development at different light intensities and the mechanisms of ABGS formation treating synthetic domestic wastewater.

Characterization of extracellular polysaccharide/protein contents during the adsorption of Cd(II) by Synechocystis sp. PCC6803.

Cyanobacteria have been proven to be cheaper and more effective for the removal of metallic elements in aqueous solutions. In this study, the living cyanobacteria Synechocystis sp. PCC6803 was used to adsorb Cd(II) and its extracellular polymeric substances (EPS) were investigated in the adsorption process. The initial stage of adsorption of Cd(II) was a rapid process, and then increase slowly accompanied with the increases of biomass. The final adsorption percentage could achieve 86% when the Cd(II) concentration was 0.5mg/L. It proved that Synechocystis sp. PCC6803 has a good adsorption capacity for heavy metal ions. EPS was extracted to investigate the secretion of which was dynamic and the maximum extracellular polysaccharides and proteins were 134.2 and 100.9mg/g, respectively. Furthermore, the real-time PCR (RT-PCR) results of genes (slr0977 and exoD) involved in EPS synthesis and secretion indicated that the EPS production was firstly increased and then decreased slightly. Transmission electron microscope (TEM) observation revealed that heavy metal ions were absorbed into EPS layer. Fourier transform infrared spectrum (FT-IR) analysis showed that EPS was rich in functional groups which could combine with heavy metal ions, such as -OH and -NH groups. All the results obtained show that the secretion of EPS by cyanobacteria was one of the ways to resist heavy metal stress. And it shows a trend of rising first and then decreasing, the change regulation of which was consistent with adsorptive behavior.

EXTRACELLULAR POLYMERIC SUBSTANCE (EPS) FROM KOCURIA SP. BRI 36: A KEY COMPONENT IN HEAVY METAL RESISTANCE

Objective: Evaluation of Extracellular Polymeric Substance (EPS) induced heavy metal tolerance in Kocuria sp. BRI 36.Methods: Initially, the effect of different concentrations of glucose (1-10 %) on EPS production by BRI 36 was examined. At optimum glucose concentrations, EPS levels were measured by varying heavy metal concentrations (10-50 ppm) of Pb2+, Cd 2+and Cr3+. Maximum tolerable concentration (MTC) and survival percentage of BRI 36 were determined under conditions that support EPS synthesis. Comparative analysis of extracted crude EPS was performed by Fourier Transform Infrared Spectroscopy (FTIR) to establish functional groups involved in the metal interaction. Results: Kocuriasp. BRI 36 produced maximum EPS (1g/l) at 5% glucose. Increase in EPS production up to 89% (considering 1g/l as 100%) with an increase in concentrations of heavy metals up to 40 ppm. MTC levels of BRI 36 for heavy metals increased up to 700 ppm when it was cultivated in presence of 5% glucose indicating a major role of surface polymer in metal adsorption. The function of EPS as a protective cover was also evident from an increase in survival percentage of BRI 36 up to 39.4 at MTC. Comparative analysis of extracted crude EPS by FTIR revealed the involvement of O-H, C=O, and C-O-C groups in metal adsorptionConclusion: Antarctic oceanic isolate Kocuria sp. BRI 36 has an ability to produced EPS under stress conditions of heavy metals. Simultaneously, its MTC values increased due to increase in EPS levels. These observations suggest the possibility to develop gentle, environmentally safe and cost-effective method for heavy metal removal.

Iron uptake and biofilm formation in Pseudomonas aeruginosa

Biofilms are surface-associated communities of microorganisms embedded within self-secreted extracellular polymeric substances, and a major cause of chronic and persistent infections. Respiratory Pseudomona aeruginosa infection is the leading reason for morbidity and mortality in cystic fibrosis patients. The formation of biofilms by P. aeruginosa in the airway is thought to increase persistence and antibiotic resistance during infection. Biofilm formation of P. aeruginosa is regulated by complicated signaling systems including quorum sensing and two-component systems that control the synthesis of extracellular polymeric substances. Furthermore, iron is an essential and scarce nutrient for bacteria and an important signal factor. P. aeruginosa has developed multiple iron uptake systems to sequester enough iron for its survival, with important regulatory roles in both release of virulence factors and formation of biofilms. In this review, we summarize recent advances in biofilm formation and its regulation along with the iron-uptake strategies in P. aeruginosa, to provide new insights and understanding to fight bacterial biofilms.

The diversity, distribution and function of N-acyl-homoserine lactone (AHL) in industrial anaerobic granular sludge

Although AHL-mediated quorum sensing (QS) signaling has been proved to be ecologically important in biofilm formation and aerobic granulation process, the biological role of AHL in anaerobic granule has not been experimentally investigated. In this paper, we explored the AHL level in 10 full-scale industrial anaerobic granular bioreactors and detected a total of 4 kinds of AHLs. C8-HSL and C10-HSL were indicated to be the potentially universal QS signal molecules in anaerobic granules and involved in extracellular polymeric substance (EPS) production and granulation process. The add-back experiments further supported the hypothesis that C8-HSL and C10-HSL might play an important role in facilitating anaerobic granulation through regulation of EPS synthesis. 12 Microorganisms might be regulated by AHL to play an important role in EPS production. This study provides a foundation for exploring the function of AHL in anaerobic granular systems, which may shed light on advanced AHL-based anaerobic granulation strategy.

EPS Characterization of a Cell Wall-Lacking Archaeon &lt;i&gt;Ferroplasma&lt;/i&gt; &lt;i&gt;acidiphilum&lt;/i&gt;

We studied the surface properties of F. acidiphilum DSM 28986 by attenuated total reflection-Fourier transformed infra-red (ATR-FTIR) spectroscopy and microbial adhesion to hydrocarbon (MATH) techniques. In addition, extracellular polymeric substances (EPS) were extracted and characterized by conventional colorimetric analysis and fluorescence lectin-binding analysis (FLBA). Results showed that: 1) cells selectively adhered to mineral surfaces and showed maximum attachment to pyrite of approx. 50% within 30 min; 2) EPS synthesis by F. acidiphilum DSM 28986 was influenced by growth substrates; and 3) tightly-bound EPS (capsular EPS) were composed of carbohydrates and proteins. In contrast, loosely-bound EPS (colloidal EPS) were mainly characterized as carbohydrates. Monosaccharides like glucose, fucose, arabinose, galactose, mannose, and sialic acid were detected in the EPS of F. acidiphilum DSM 28986. This study provides first insight into surface characterization of the cell wall-lacking archaeon F. acidiphilum and facilitates the understanding of interactions of this organism with other acidophiles and metal sulfides.

The effects of substrate exposure history and carbon starvation-induced stress on the EPS synthesis of TCE degrading toluene oxidizing soil bacteria

A series of batch experiments was undertaken to explore the influence of growth conditions on the extracellular polymeric substances (EPS) production of five representative strains of trichloroethylene (TCE) degrading toluene oxidizing bacteria of importance to in situ bioremediation. EPS production for three distinct carbon sources (i.e., lactate, glucose, and toluene) was examined under starvation and non-starvation conditions. EPS extraction was accomplished using DOWEX™ (cation exchange resin; CER), and total carbohydrate, protein, and uronic acid were quantified as the main EPS constituents. Experimental results reveal that most bacteria grown on toluene synthesized more EPS per colony forming unit (CFU) than those grown on other substrates and that EPS production is enhanced with increasing length of starvation. These observations suggest that EPS may be produced under conditions of environmental stress and may be linked to enzyme deactivation. The results from this work additionally indicate that the mutant strain of Ralstonia pickettii PKO1 with the lack of gene encoding TbuX (an outer membrane protein) is more hydrophobic due to less EPS synthesis ability. The relatively lower amount of EPS synthesis in the mutant strain of R. pickettii PKO1 suggests that TbuX, an outer membrane protein encoded a part of the toluene catabolic regulation of R. pickettii PKO1, has a role in bacterial EPS production.

Heavy metals removal from wastewater using extracellular polymeric substances produced by Cloacibacterium normanense in wastewater sludge supplemented with crude glycerol and study of extracellular polymeric substances extraction by different methods

Extracellular polymeric substances synthesis by Cloacibacterium was affected by different concentrations of glycerol in the medium. The concentration of EPS in 72h fermentation was increased from 13g/L with no external carbon supplementation to 21.3±0.7g/L with 2% (w/v) crude glycerol addition. Physical and chemical extraction methods (heating, centrifugation and ethylene diamine tetra-acetic acid (EDTA)) were used in this study and their performance to extract EPS was compared. A significant variation in concentration of extracted B-EPS (broth-EPS) by heating (20.8±0.5g/L) and centrifugation (21.3±0.7g/L) extraction methods was not observed. However, in case of extraction with EDTA (5g/L), the B-EPS concentration extracted was 25.5±0.9g/L, which exhibited high flocculation activity of 95.3±0.5% at optimum dose of 23.1mgB-EPS/gkaolin. Moreover, Ni removal efficiency of 80% from primary treated wastewater was achieved using 35mg/L of B-EPS extracted by centrifugation method.

The effect of quorum sensing on anaerobic granular sludge in different pH conditions

The effects of several signal molecules on anaerobic granular sludge at different pHs were investigated. The results showed that pH affected the synthesis and maintaining of the signal molecules. In an alkaline medium (pH 9), sludge bacteria secreted less acyl-homoserine lactone (AHLs), which had an opposite variation with the synthesis of extracellular polymeric substances (EPS), and its effect did not present. Additionally, the synthesis of EPS (mainly loosely bound layer) was majorly promoted by auto-inducers-2 (AI-2), but a lower negative role of diffusible signal factor (DSF). In neutral and weak alkaline media, the combining regulation of increasing AI-2 content and decreasing DSF content enhanced relative hydrophobicity (RH) and granular strength in granular sludge, which made granules to have a greater diameter and a more glossy granular shape. Whereas, in acid medium, the decreasing AI-2 content and increasing DSF content made a decrease in RH and granular strength, resulting in the degeneration of anaerobic granular sludge.

Chemical Biology Strategies for Biofilm Control.

Microbes live as densely populated multicellular surface-attached biofilm communities embedded in self-generated, extracellular polymeric substances (EPSs). EPSs serve as a scaffold for cross-linking biofilm cells and support development of biofilm architecture and functions. Biofilms can have a clear negative impact on humans, where biofilms are a common denominator in many chronic diseases in which they prime development of destructive inflammatory conditions and the failure of our immune system to efficiently cope with them. Our current assortment of antimicrobial agents cannot efficiently eradicate biofilms. For industrial applications, the removal of biofilms within production machinery in the paper and hygienic food packaging industry, cooling water circuits, and drinking water manufacturing systems can be critical for the safety and efficacy of those processes. Biofilm formation is a dynamic process that involves microbial cell migration, cell-to-cell signaling and interactions, EPS synthesis, and cell-EPS interactions. Recent progress of fundamental biofilm research has shed light on novel chemical biology strategies for biofilm control. In this article, chemical biology strategies targeting the bacterial intercellular and intracellular signaling pathways will be discussed.

Insights into the effect of mixed engineered nanoparticles on activated sludge performance.

In this study, the effects, fate and transport of ENPs in wastewater treatment plants (WWTP) were investigated using three parallel pilot WWTPs operated under identical conditions. The WWTPs were spiked with (i) an ENP mixture consisting of silver oxide, titanium dioxide and zinc oxide, and (ii) bulk metal salts. The third plant served as control (unspiked). ENP effects were evaluated for (i) bulk contaminant removal, (ii) activated sludge (AS) process performance, (iii) microbial community structure and dynamics and (iv) microbial inhibition. ENPs showed a strong affinity for biosolids and induced a specific oxygen uptake rate two times higher than the control. The heterotrophic biomass retained its ability to nitrify and degrade organic matter. However, non-recovery of ammonia- and nitrite-oxidizing bacteria such as Nitrosomonas, Nitrobacter or Nitrospira in the ENP spiked reactors suggests selective inhibitory effects. The results further suggest that ENPs and metal salts have antimicrobial properties which can reduce synthesis of extracellular polymeric substances and therefore floc formation. Scanning electron microscopy evidenced selective damage to some microbes, whereas lipid fingerprinting and 454 pyrosequencing indicated a temporal shift in the microbial community structure and diversity. Acidovorax, Rhodoferax, Comamonas and Methanosarcina were identified as nano-tolerant species. Competitive growth advantage of the nano-tolerant species influenced the removal processes and unlike other xenobiotic compounds, ENPs can hasten the natural selection of microbial species in AS.