Swarming Motility Research Articles

In Silico and In Vitro Screening of Antipathogenic Properties of Melianthus comosus (Vahl) against Pseudomonas aeruginosa.

Bacterial quorum sensing (QS) system regulates pathogenesis, virulence, and biofilm formation, and together they contribute to nosocomial infections. Opportunistic pathogens, such as Pseudomonas aeruginosa, rely on QS for regulating virulence factors. Therefore, blocking the QS system may aid management of various infectious diseases caused by human pathogens. Plant secondary metabolites can thwart bacterial colonization and virulence. As such, this study was undertaken to evaluate three extracts from the medicinal plant, Melianthus comosus, from which phytochemical compounds were identified with potential to inhibit QS-dependent virulence factors in P. aeruginosa. Chemical profiling of the three extracts identified 1,2-benzene dicarboxylic acid, diethyl ester, neophytadiene and hexadecanoic acid as the common compounds. Validation of antibacterial activity confirmed the same MIC values of 0.78 mg/mL for aqueous, methanol and dichloromethane extracts while selected guanosine showed MIC 0.031 mg/mL. Molecular docking analysis showed anti-quorum sensing (AQS) potential of guanosine binding to CviR’ and 2UV0 proteins with varying docking scores of −5.969 and −8.376 kcal/mol, respectively. Guanosine inhibited biofilm cell attachment and biofilm development at 78.88% and 34.85%, respectively. Significant swimming and swarming motility restriction of P. aeruginosa were observed at the highest concentration of plant extracts and guanosine. Overall, guanosine revealed the best swarming motility restrictions. M. comosus extracts and guanosine have shown clear antibacterial effects and subsequent reduction of QS-dependent virulence activities against P. aeruginosa. Therefore, they could be ideal candidates in the search for antipathogenic drugs to combat P. aeruginosa infections.

Effects of Gentamicin-Loaded Chitosan-ZnO Nanocomposite on Quorum-Sensing Regulation of Pseudomonas Aeruginosa.

Cell density-based intercellular signaling mechanism is known as Quorum sensing (QS); it serves a significant role in regulating the pathogenic factors. The objective of the present study was to assess the influence of chitosan-zinc oxide nanocomposite (CH-ZnO nanocomposite), alone and in combination with gentamicin, on the sensitivity to hydrogen peroxide (H2O2), the production of pathogenic factors and QS-regulated genes of Pseudomonas aeruginosa. The efficacy of the minimum inhibitory concentration (MIC) and 1/4 MIC of the CH-ZnO nanocomposite, alone and in combination with gentamicin, on the sensitivity to H2O2, pyocyanin secretion, swarming and twitching motilities was evaluated. In addition, the expression of some QS-regulated genes including rhlI, rhlR, lasI and lasR genes was measured by Real-time quantitative PCR (RT-qPCR) following exposure to the nanocomposite. The results demonstrated that at MIC concentrations, the gentamicin-loaded CH-ZnO nanocomposite significantly inhibited QS-regulated phenotypes such as pyocyanin secretion (82.4%), swarming (76%) and twitching (73.6%) motilities; further it increased the inhibition growth zone (134.5%), as well as, at 1/4 MIC concentration decreased the expression of lasI (72%), lasR (78%), rhlI (76%) and rhlR (82%) genes; as compared to untreated P. aeruginosa PAO1 (P < 0.05). Our results also demonstrated that the CH-ZnO nanocomposite combined with gentamicin could be a potential innovative candidate, which could be broadly applied in the treatment of P. aeruginosa infections.

Inhibition of swarming motility using in vitro hyperthermia.

Hyperthermia is a therapeutic technique in which body tissue is exposed to temperatures in the region of 40-45 °C to induce a physiological or biological effect. Swarming motility is an important virulence factor for Proteus mirabilis and Pseudomonas aeruginosa and swarming phenomenon is a coordinated multicellular movement of differentiated bacterial population over semi-solid surfaces. In this study, we aimed to investigate the inhibitory effect of hyperthermia on bacterial swarming motility using a modified thermobiogram method and show the potential of this thermal method to treat bacterial infections. Ten P.mirabilis and 10 P.aeruginosa clinical isolates were included in the study. Sheep blood agar (SBA) plates were prepared and inoculated with bacterial suspensions of clinical isolates. Inoculated SBA plates were incubated inside 2 different incubators; at 37 °C and 45 °C for 20 h. The diameter of bacterial growing zones (swarming diameters) were measured every 2 h and noted. Finally, Gram stains of the isolates were prepared for microscopic examination. Wilcoxon signed-rank test was used to compare the swarming inhibition rates of the isolates incubated at 37 °C and 45 °C. Regarding P.mirabilis species, a significant difference was found between two different temperatures (P = 0.0078). So, a temperature at the level of hyperthermia significantly inhibited the swarming motility of P.mirabilis isolates. In addition, transformation to coccus form was observed at 45 °C. We speculate that these findings might be useful for employing thermal therapies including hyperthermia method to treat infectious diseases caused by swarming bacterial pathogens in the future.

Peptide 1018 inhibits swarming and influences Anr-regulated gene expression downstream of the stringent stress response in Pseudomonas aeruginosa

Pseudomonas aeruginosa is a ubiquitous opportunistic pathogen that causes considerable human morbidity and mortality, particularly in nosocomial infections and individuals with cystic fibrosis. P. aeruginosa can adapt to surface growth by undergoing swarming motility, a rapid multicellular movement that occurs on viscous soft surfaces with amino acids as a nitrogen source. Here we tested the small synthetic host defense peptide, innate defense regulator 1018, and found that it inhibited swarming motility at concentrations as low as 0.75 μg/ml, well below the MIC for strain PA14 planktonic cells (64 μg/ml). A screen of the PA14 transposon insertion mutant library revealed 29 mutants that were more tolerant to peptide 1018 during swarming, five of which demonstrated significantly greater swarming than the WT in the presence of peptide. Transcriptional analysis (RNA-Seq) of cells that were inoculated on swarming plates containing 1.0 μg/ml peptide revealed differential expression of 1,190 genes compared to cells swarming on plates without peptide. Furthermore, 1018 treatment distinctly altered the gene expression profile of cells when compared to that untreated cells in the centre of the swarm colonies. Peptide-treated cells exhibited changes in the expression of genes implicated in the stringent stress response including those regulated by anr, which is involved in anaerobic adaptation, indicative of a mechanism by which 1018 might inhibit swarming motility. Overall, this study illustrates potential mechanisms by which peptide 1018 inhibits swarming surface motility, an important bacterial adaptation associated with antibiotic resistance, virulence, and dissemination of P. aeruginosa.

Antivirulence Properties of a Low-Molecular-Weight Quaternized Chitosan Derivative against Pseudomonas aeruginosa.

The co-occurrence of increasing rates of resistance to current antibiotics and the paucity of novel antibiotics pose major challenges for the treatment of bacterial infections. In this scenario, treatments targeting bacterial virulence have gained considerable interest as they are expected to exert a weaker selection for resistance than conventional antibiotics. In a previous study, we demonstrated that a low-molecular-weight quaternized chitosan derivative, named QAL, displays antibiofilm activity against the major pathogen Pseudomonas aeruginosa at subinhibitory concentrations. The aim of this study was to investigate whether QAL was able to inhibit the production of relevant virulence factors of P. aeruginosa. When tested in vitro at subinhibiting concentrations (0.31–0.62 mg/mL), QAL markedly reduced the production of pyocyanin, pyoverdin, proteases, and LasA, as well as inhibited the swarming motility of three out of four P. aeruginosa strains tested. Furthermore, quantitative reverse transcription PCR (qRT-PCR) analyses demonstrated that expression of lasI and rhlI, two QS-related genes, was highly downregulated in a representative P. aeruginosa strain. Confocal scanning laser microscopy analysis suggested that FITC-labelled QAL accumulates intracellularly following incubation with P. aeruginosa. In contrast, the reduced production of virulence factors was not evidenced when QAL was used as the main polymeric component of polyelectrolyte-based nanoparticles. Additionally, combination of sub-MIC concentrations of QAL and tobramycin significantly reduced biofilm formation of P. aeruginosa, likely due to a synergistic activity towards planktonic bacteria. Overall, the results obtained demonstrated an antivirulence activity of QAL, possibly due to polymer intracellular localization and QS-inhibition, and its ability to inhibit P. aeruginosa growth synergizing with tobramycin.

Bacterial-induced pH shifts link individual cell physiology to macroscale collective behavior

Bacteria have evolved a diverse array of signaling pathways that enable them to quickly respond to environmental changes. Understanding how these pathways reflect environmental conditions and produce an orchestrated response is an ongoing challenge. Herein, we present a role for collective modifications of environmental pH carried out by microbial colonies living on a surface. We show that by collectively adjusting the local pH value, Paenibacillus spp., specifically, regulate their swarming motility. Moreover, we show that such pH-dependent regulation can converge with the carbon repression pathway to down-regulate flagellin expression and inhibit swarming in the presence of glucose. Interestingly, our results demonstrate that the observed glucose-dependent swarming repression is not mediated by the glucose molecule per se, as commonly thought to occur in carbon repression pathways, but rather is governed by a decrease in pH due to glucose metabolism. In fact, modification of the environmental pH by neighboring bacterial species could override this glucose-dependent repression and induce swarming of Paenibacillus spp. away from a glucose-rich area. Our results suggest that bacteria can use local pH modulations to reflect nutrient availability and link individual bacterial physiology to macroscale collective behavior.

Characterization of the relationship between polar and lateral flagellar genes in clinical Aeromonas dhakensis: phenotypic, genetic and biochemical analyses

Flagellar-mediated motility is a crucial virulence factor in many bacterial species. A dual flagellar system has been described in aeromonads; however, there is no flagella-related study in the emergent human pathogen Aeromonas dhakensis. Using 46 clinical A. dhakensis, phenotypic motility, genotypic characteristics (flagellar genes and sequence types), biochemical properties and their relationship were investigated in this study. All 46 strains showed swimming motility at 30 °C in 0.3% Bacto agar and carried the most prevalent 6 polar flagellar genes cheA, flgE, flgG, flgH, flgL, and flgN. On the contrary, only 18 strains (39%) demonstrated swarming motility on 0.5% Eiken agar at 30 °C and they harbored 11 lateral flagellar genes lafB, lafK, lafS, lafT, lafU, flgCL, flgGL, flgNL, fliEL, fliFL, and fliGL. No association was found between biochemical properties and motility phenotypes. Interestingly, a significant association between swarming and strains isolated from pus was observed (p = 0.0171). Three strains 187, 277, and 289 isolated from pus belonged to novel sequence types (ST522 and ST524) exhibited fast swimming and swarming profiles, and they harbored > 90% of the flagellar genes tested. Our findings provide a fundamental understanding of flagellar-mediated motility in A. dhakensis.

(+)-Terpinen-4-ol Inhibits Bacillus cereus Biofilm Formation by Upregulating the Interspecies Quorum Sensing Signals Diketopiperazines and Diffusing Signaling Factors.

Bacillus cereus is a Gram-positive endospore-forming foodborne pathogen that causes lethal food poisoning and significant economic losses, usually through biofilm- and endospore-induced recurrent cross- and postprocessing contamination. Due to the lack of critical inhibitory targets and control strategies, B. cereus biofilm contamination is a problem that urgently needs a solution. In this study, the antibacterial and antibiofilm activities of several natural potential bacterial quorum sensing (QS) interferers, a group of spice-originated monoterpenoids, were screened, and terpinen-4-ol effectively inhibited B. cereus growth and biofilm and spore germination with minimum growth inhibition and 50% biofilm inhibitory concentrations of 8 and 2 μmol/mL, respectively. FESEM/CLSM and phenotypic research illustrated that in addition to a decrease in the number of attached B. cereus cells, (+)-terpinen-4-ol also obviously reduced extracellular matrix synthesis, especially exopolysaccharides, and inhibited the swarming motility and protease activity of B. cereus. (+)-Terpinen-4-ol did not exert a significant effect on AI-2 signals in B. cereus. Accordingly, the B. cereus-produced interspecies QS signals diffusing signal factors (DSFs, C8-C15) and diketopiperazines (DKPs) were detected and identified here, which suppressed B. cereus biofilm formation in a concentration-dependent manner. (+)-Terpinen-4-ol significantly increased the levels of specific DSF and DKP signals in B. cereus and down-regulated the gene expression of some rpfB homologues in transcription level. Moreover, both DKPs and DSFs inhibited swarming motility and protease activity in B. cereus, while just the DSF signals 2-dodecenoic acid and 11-methyl-2-dodecenoic acid inhibited exopolysaccharide synthesis like (+)-terpinen-4-ol. In summary, B. cereus strains were found to produce nine DSF- and six DKP-type QS signaling molecules, which repressed B. cereus biofilm formation. (+)-Terpinen-4-ol was confirmed to be a promising antibacterial and antibiofilm agent against B. cereus upregulating DSFs and DKPs levels, and it could target the critical genes rpfB for DSFs turnover.

Glucose-6-Phosphate Acts as an Extracellular Signal of SagS To Modulate Pseudomonas aeruginosa c-di-GMP Levels, Attachment, and Biofilm Formation.

ABSTRACTIn Pseudomonas aeruginosa, the orphan two-component sensor SagS contributes both to transition to biofilm formation and to biofilm cells gaining their heightened tolerance to antimicrobials. However, little is known about the identity of the signals or conditions sensed by SagS to induce the switch to the sessile, drug-tolerant mode of growth. Using a modified Biolog phenotype assay to screen for compounds that modulate attachment in a SagS-dependent manner, we identified glucose-6-phosphate to enhance attachment in a manner dependent on the glucose-6-phosphate concentration and SagS. The stimulatory effect was not limited to the attachment since glucose-6-phosphate likewise enhanced biofilm formation and also enhanced the expression of select biofilm marker genes. Moreover, exposure to glucose-6-phosphate coincided with decreased swarming motility but increased cellular cyclic-di-GMP (c-di-GMP) levels in biofilms. No such response was noted for compounds modulating attachment and biofilm formation in a manner independent of SagS. Modulation of c-di-GMP in response to glucose-6-phosphate was due to the diguanylate cyclase NicD, with NicD also being required for enhanced biofilm formation. The latter was independent of the sensory domain of NicD but dependent on NicD activity, SagS, and the interaction between NicD and SagS. Our findings indicate that glucose-6-phosphate likely mimics a signal or conditions sensed by SagS to activate its motile-sessile switch function. In addition, our findings provide new insight into the interfaces between the ligand-mediated two-component system signaling pathway and c-di-GMP levels.IMPORTANCE Pathogens sense and respond to signals and cues present in their environment, including host-derived small molecules to modulate the expression of their virulence repertoire. Here, we demonstrate that the opportunistic pathogen Pseudomonas aeruginosa responds to glucose-6-phosphate. Since glucose-6-phosphate is primarily made available due to cell lysis, it is likely that glucose-6-phosphate represents a cross-kingdom cell-to-cell signal that enables P. aeruginosa to adapt to the (nutrient-poor) host environment by enhancing biofilm formation, cyclic-di-GMP, and the expression of genes linked to biofilm formation in a concentration- and SagS-dependent manner.

Comparing bacterial properties in relation to the virulence factors of Xanthomonas citri subsp. citri strains and evaluating resistance of subtribe Citrinae cultivars to the most virulent strain

Citrus Bacterial Canker disease, caused by Xanthomonas citri subsp. citri (Xcc), is one of the most devastating diseases that attacks citrus, especially in the southern areas of Iran. The objective of this investigation was to analyze several characteristics involved in Xcc virulence in relation to strain aggressiveness. To achieve this, swarming motility, biofilm formation, resistance to H2O2, and production of xanthan were evaluated considering 44 local strains causing citrus bacterial canker disease in Iran. All strains showed differential swarming motilities, biofilm formation abilities, and xanthan production. The most virulent strain, Xcc-KVXCC1, exhibited the greatest capability to form biofilm on solid surfaces, xanthan production, and pathogenicity on detached leaves of Citrus aurantifolia as well as positive chemotaxis toward C. aurantifolia extract. In this study, a total of 14 common and commercial citrus genotypes were evaluated for resistance to Xcc-KVXCC1. Genotypes were categorized into susceptible and resistant groups based on lesion number per inoculation site. Based on our results, C. aurantifolia was the most susceptible citrus cultivar to Xcc-KVXCC1. In contrast, C. aurantium, C. jumbhori, and C. reticulata cv. ponkan demonstrated high and moderately high resistance against Xcc-KVXCC1.

New insights into the antibacterial and quorum sensing inhibition mechanism of Artemisia argyi leaf extracts towards Pseudomonas aeruginosa PAO1.

The online version contains supplementary material available at 10.1007/s13205-021-02663-5.

Feed supplementation with quorum quenching probiotics with anti-virulence potential improved innate immune responses, antioxidant capacity and disease resistance in Asian seabass (Lates calcarifer)

Application of multifunctional probiotics is considered to be a practical strategy for the control of infections caused by Gram-negative bacteria. Consequently, the present study was designed to investigate the possible inhibitory effects of quorum quenching (QQ) probiotics on several N-acyl-homoserine lactone (AHL)-associated virulence factors in Vibrio alginolyticus and V. harveyi in vitro, and to evaluate the impacts of their administration in the diet on non-specific immune responses, antioxidant capacity and resistance to V. alginolyticus in Asian seabass. For the in vitro assay, the effect of exogenous AHL was assayed on the regulation of several virulence factors including lytic enzymes (amylase, gelatinase, hemolysin, lecithinase, lipase and protease productions), swarming and swimming motilities, and biofilm formation in V. alginolyticus and V. harveyi. Exogenous 3-oxo-C10-HSL positively regulated the production of amylase, gelatinase, protease and biofilm in V. alginolyticus. Exogenous 3-oh-C4-HSL positively regulated swarming, swimming and biofilm formation in V. harveyi. To determine the virulence-inhibitory effects of QQ probiotics on the AHL-dependent virulence factors, cell-free lysates were obtained from Bacillus thuringiensis QQ1 and B. cereus QQ2. Both probionts could reduce the production of amylase, gelatinase and protease in V. alginolyticus. Bacillus thuringiensis QQ1 was also able to decrease the biofilm formed by V. alginolyticus. Bacillus thuringiensis QQ1 and B. cereus QQ2 decreased swarming and swimming motilities, and biofilm formation in V. harveyi. In order to assay the effectiveness of the QQ probiotics in vivo, Asian seabass were fed with basal diet (control) and basal diet supplemented with 109 CFU/g feed of B. thuringiensis QQ1 or B. cereus QQ2 for 8 weeks. At the end of pre-infection trial (day 42), catalase (CAT), superoxide dismutase (SOD), lysozyme, antiprotease, myeloperoxidase (MPO), respiratory burst (RB) and bactericidal activities were significantly (p < 0.05) higher in the probiotic-treated fish than un-treated fish. After challenge with V. alginolyticus, significantly (p < 0.05) higher levels of glutathione (GSH) in the QQ2 treatment (day 49) and lower levels of malondialdehyde (MDA) in both probiotic groups were observed compared to the control group. Moreover, CAT, MPO, RB, alternative pathway of complement (APC), lysozyme, antiprotease and bactericidal activities were significantly (p < 0.05) higher in the probiotic treatments compared to the control group after 7 and 14 days post-infection. Both probionts significantly (p < 0.05) improved the cumulative survival in Asian seabass against V. alginolyticus. Our results indicate that QQ probiotics can modulate immune and antioxidant responses, and provide high protection against Vibrio infection in Asian seabass.

In vitro activity of hyperthermia on swarming motility and antimicrobial susceptibility profiles of Proteus mirabilis isolates

Aim Swarming motility is a virulence factor for Proteus mirabilis and is a coordinated multicellular movement of bacteria. In this study, we investigated the inhibitory effect of hyperthermia on bacterial swarming motility and antimicrobial resistance. Methods Thirty-one P. mirabilis isolates were included in the study. Seven inoculated agar plates were incubated inside incubators with increasing temperature levels: at 36 °C (control) and 40–45 °C. On the next day, inhibition of swarming was evaluated and minimum paralyzing temperature (MPT) values were determined. An antimicrobial susceptibility test (antibiogram) is performed by exposing bacteria to increasing concentrations of antibiotics, in vitro. Thus, we used the Kirby-Bauer disk diffusion test as a screening method to analyze the antibiogram profiles of the isolates at 36 °C and 42 °C. Finally, a time-kill assay was performed to analyze the killing effect of hyperthermia (42 °C) on planktonic bacteria, in combination with the antibiotic meropenem at the first and third hours. A Wilcoxon signed-rank test was used to compare the killing effects of meropenem, hyperthermia and their combinations. Results The median MPT value was determined as 44 °C. In the disk diffusion assay, susceptibility development was observed in 94% of isolates for at least one antibiotic. In the time-kill assay, we observed a significant killing effect of hyperthermia in combination with meropenem. Under the microscope, we observed the formation of spherical cells by the effect of heat. Conclusion We conclude that these findings might be useful when employing the hyperthermia method to treat infectious diseases caused by P. mirabilis in the future.

WbaP is required for swarm motility and intramacrophage multiplication of Salmonella Enteritidis spiC mutant by glucose use ability

Salmonella spp. can survive and replicate in macrophage cells to cause persistent infection, SpiC is a necessary T3SS effector, but its pathogenic mechanism is still not known completely. In our study, Salmonella Enteritidis spiC mutant (SEΔspiC) was found to have stronger swarming motility and intramacrophage hyperproliferation which was closely related to glucose metabolism. SEΔspiC wbaP::Tn5 mutant was screened out by transposon mutagenesis, which had weaker swarming motility and intramacrophage replication ability than SEΔspiC in the presence of glucose. Bioinformatics displayed that undecaprenyl-phosphate galactose phosphotransferase (Wbap), encoded by wbaP gene, was a key enzyme for glucose metabolism and Lipopolysaccharide(LPS) synthesis, which confirmed our outcome that Wbap was involved in intramacrophage replication ability by glucose use in addition to swarming motility based on SEΔspiC. This discovery will further promote the understanding of the interaction between wbaP gene and spiC gene and the intracellular Salmonella replication mechanism.

Contribution of Swarming Motility to Dissemination in a Pseudomonas aeruginosa Murine Skin Abscess Infection Model.

Swarming motility in Pseudomonas aeruginosa is a multicellular adaptation induced by semisolid medium with amino acids as a nitrogen source. By phenotypic screening, we differentiated swarming from other complex adaptive phenotypes, such as biofilm formation, swimming and twitching, by identifying a swarming-specific mutant in ptsP, a metabolic regulator. This swarming-deficient mutant was tested in an acute murine skin abscess infection model. Bacteria were recovered at significantly lower numbers from organs of mice infected with the ∆ptsP mutant. We also tested the synthetic peptide 1018 for activity against different motilities and efficacy in vivo. Treatment with peptide 1018 mimicked the phenotype of the ∆ptsP mutant in vitro, as swarming was inhibited at low concentrations (<2 μg/mL) but not swimming or twitching, and in vivo, as mice had a reduced bacterial load recovered from organs. Therefore, PtsP functions as a regulator of swarming, which in turn contributes to dissemination and colonization in vivo.

Bacterial tRNA 2'-O-methylation is dynamically regulated under stress conditions and modulates innate immune response.

RNA modifications are a well-recognized way of gene expression regulation at the post-transcriptional level. Despite the importance of this level of regulation, current knowledge on modulation of tRNA modification status in response to stress conditions is far from being complete. While it is widely accepted that tRNA modifications are rather dynamic, such variations are mostly assessed in terms of total tRNA, with only a few instances where changes could be traced to single isoacceptor species. Using Escherichia coli as a model system, we explored stress-induced modulation of 2′-O-methylations in tRNAs by RiboMethSeq. This analysis and orthogonal analytical measurements by LC-MS show substantial, but not uniform, increase of the Gm18 level in selected tRNAs under mild bacteriostatic antibiotic stress, while other Nm modifications remain relatively constant. The absence of Gm18 modification in tRNAs leads to moderate alterations in E. coli mRNA transcriptome, but does not affect polysomal association of mRNAs. Interestingly, the subset of motility/chemiotaxis genes is significantly overexpressed in ΔTrmH mutant, this corroborates with increased swarming motility of the mutant strain. The stress-induced increase of tRNA Gm18 level, in turn, reduced immunostimulation properties of bacterial tRNAs, which is concordant with the previous observation that Gm18 is a suppressor of Toll-like receptor 7 (TLR7)-mediated interferon release. This documents an effect of stress induced modulation of tRNA modification that acts outside protein translation.

Rampant loss of social traits during domestication of a Bacillus subtilis natural isolate

Most model bacteria have been domesticated in laboratory conditions. Yet, the tempo with which a natural isolate diverges from its ancestral phenotype under domestication to a novel laboratory environment is poorly understood. Such knowledge, however is essential to understanding the rate of evolution, the time scale over which a natural isolate can be propagated without loss of its natural adaptive traits, and the reliability of experimental results across labs. Using experimental evolution, phenotypic assays, and whole-genome sequencing, we show that within a week of propagation in a common laboratory environment, a natural isolate of Bacillus subtilis acquires mutations that cause changes in a multitude of traits. A single adaptive mutational step in the gene coding for the transcriptional regulator DegU impairs a DegU-dependent positive autoregulatory loop and leads to loss of robust biofilm architecture, impaired swarming motility, reduced secretion of exoproteases, and to changes in the dynamics of sporulation across environments. Importantly, domestication also resulted in improved survival when the bacteria face pressure from cells of the innate immune system. These results show that degU is a target for mutations during domestication and underscores the importance of performing careful and extremely short-term propagations of natural isolates to conserve the traits encoded in their original genomes.

Biological role of EPS from Pseudomonas syringae pv. syringae UMAF0158 extracellular matrix, focusing on a Psl-like polysaccharide

Pseudomonas syringae is a phytopathogenic model bacterium that is used worldwide to study plant–bacteria interactions and biofilm formation in association with a plant host. Within this species, the syringae pathovar is the most studied due to its wide host range, affecting both, woody and herbaceous plants. In particular, Pseudomonas syringae pv. syringae (Pss) has been previously described as the causal agent of bacterial apical necrosis on mango trees. Pss exhibits major epiphytic traits and virulence factors that improve its epiphytic survival and pathogenicity in mango trees. The cellulose exopolysaccharide has been described as a key component in the development of the biofilm lifestyle of the P. syringae pv. syringae UMAF0158 strain (PssUMAF0158). PssUMAF0158 contains two additional genomic regions that putatively encode for exopolysaccharides such as alginate and a Psl-like polysaccharide. To date, the Psl polysaccharide has only been studied in Pseudomonas aeruginosa, in which it plays an important role during biofilm development. However, its function in plant-associated bacteria is still unknown. To understand how these exopolysaccharides contribute to the biofilm matrix of PssUMAF0158, knockout mutants of genes encoding these putative exopolysaccharides were constructed. Flow-cell chamber experiments revealed that cellulose and the Psl-like polysaccharide constitute a basic scaffold for biofilm architecture in this bacterium. Curiously, the Psl-like polysaccharide of PssUMAF0158 plays a role in virulence similar to what has been described for cellulose. Finally, the impaired swarming motility of the Psl-like exopolysaccharide mutant suggests that this exopolysaccharide may play a role in the motility of PssUMAF0158 over the mango plant surface.

Synthesis, Characterization, and Antibacterial Activity of Biosynthesized Gold Nanoparticles

In this study, Acalypha indica was utilized to green synthesize gold nanoparticles. The characteristics of the synthesized nanoparticles were observed through UV-Vis, FTIR, TEM, particle size analyzer, and XRD. Furthermore, the nanoparticles were investigated for antibacterial properties. The particle size of gold nanoparticle was around 50 – 100 nm, and the antibacterial property of the nanoparticle was assessed using agar well diffusion, swarming motility, MIC, and protein leakage assay. The gold nanoparticles were observed to be active against E .coli alone with MIC at 160 µg, and it was observed to inhibit its swarming motility and to make the cell leak out proteins.

FliC of Vibrio splendidus-related strain involved in adhesion to Apostichopus japonicus

Vibrio splendidus-related strains are important opportunistic marine pathogens, and they can infect many important marine animals, including the sea cucumber Apostichopus japonicus. In this study, one gene coding flagellin was cloned and a V. splendidus-related strain AJ01/GFPFliC with the overexpression of fliC gene was constructed to explore the function of FliC. AJ01/GFPFliC showed a 3–4 h delay in the initial growth phase and then its growth was faster than that of the wild type strain AJ01. The abilities of swarming motility and biofilm formation ability of AJ01/GFPFliC were also higher than that of AJ01. The adhesion rate of AJ01/GFPFliC to the slide and the coelomocytes of A. japonicus increased from 1% to 5%, and 25% to 40%, respectively, and the adhered AJ01/GFPFliC cells in intestinal tissue of A. japonicus reached 8.0 × 106 CFU/g, which was 2.5-fold higher than that of the control strain AJ01/GFP. Concluded from all the data suggested that FliC was an adhesion factor of V. splendidus-related strain AJ01 that could also contribute to bacterial swarming motility and biofilm formation.