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Microbiology Spectrum Jun 2024Swarming motility in pseudomonads typically requires both a functional flagellum and the production/secretion of a biosurfactant. Published work has shown that the...
Swarming motility in pseudomonads typically requires both a functional flagellum and the production/secretion of a biosurfactant. Published work has shown that the wild-type Pf0-1 is swarming deficient due to a point mutation in the gene, which until recently was thought to inactivate rather than attenuate the Gac/Rsm pathway. As a result, little is known about the underlying mechanisms that regulate swarming motility by Pf0-1. Here, we demonstrate that a Δ Δ Δ mutant, which phenotypically mimics Gac/Rsm pathway overstimulation, is proficient at swarming motility. RsmA and RsmE appear to play a key role in this regulation. Transposon mutagenesis of the Δ Δ Δ mutant identified multiple factors that impact swarming motility, including pathways involved in flagellar synthesis and biosurfactant production/secretion. We find that loss of genes linked to biosurfactant Gacamide A biosynthesis or secretion impacts swarming motility, as does loss of the alternative sigma factor FliA, which results in a defect in flagellar function. Collectively, these findings provide evidence that Pf0-1 can swarm if the Gac/Rsm pathway is activated, highlight the regulatory complexity of swarming motility in this strain, and demonstrate that the cyclic lipopeptide Gacamide A is utilized as a biosurfactant for swarming motility.IMPORTANCESwarming motility is a coordinated process that allows communities of bacteria to collectively move across a surface. For Pf0-1, this phenotype is notably absent in the parental strain, and to date, little is known about the regulation of swarming in this strain. Here, we identify RsmA and RsmE as key repressors of swarming motility modulating the levels of biosurfactant production/secretion. Using transposon mutagenesis and subsequent genetic analyses, we further identify potential regulatory mechanisms of swarming motility and link Gacamide A biosynthesis and transport machinery to swarming motility.
Topics: Pseudomonas fluorescens; Movement; Bacterial Proteins; Methyltransferases; Surface-Active Agents; Mutagenesis; Sigma Factor
PubMed: 38687073
DOI: 10.1128/spectrum.00166-24 -
Frontiers in Microbiology 2023Bacterial communication is a fundamental process used to synchronize gene expression and collective behavior among the bacterial population. The most studied bacterial...
Bacterial communication is a fundamental process used to synchronize gene expression and collective behavior among the bacterial population. The most studied bacterial communication system is quorum sensing, a cell density system, in which the concentration of inductors increases to a threshold level allowing detection by specific receptors. As a result, bacteria can change their behavior in a coordinated way. While in quorum sensing based on the synthesis of -acyl homoserine lactone molecules is well studied, volatile organic compounds, although considered to be communication signals in the rhizosphere, are understudied. The MFE01 strain has a very active type six secretion system that can kill some competitive bacteria. Furthermore, MFE01 emits numerous volatile organic compounds, including 1-undecene, which contributes to the aerial inhibition of growth. Finally, MFE01 appears to be deprived of -acyl homoserine lactone synthase. The main objective of this study was to explore the role of 1-undecene in the communication of MFE01. We constructed a mutant affected in gene encoding the enzyme responsible for 1-undecene synthesis to provide further insight into the role of 1-undecene in MFE01. First, we studied the impacts of this mutation both on volatile organic compounds emission, using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry and on long-range inhibition. Then, we analyzed influence of 1-undecene on MFE01 coordinated phenotypes, including type six secretion system activity and biofilm formation. Next, to test the ability of MFE01 to synthesize -acyl homoserine lactones in our conditions, we investigated the presence of corresponding genes across the MFE01 genome and we exposed its biofilms to an acyl homoserine lactone-degrading enzyme. Finally, we examined the effects of 1-undecene emission on MFE01 biofilm maturation and aerial communication using an original experimental set-up. This study demonstrated that the Δ mutant is impaired in biofilm maturation. An exposure of the Δ mutant to the volatile compounds emitted by MFE01 during the biofilm development restored the biofilm maturation process. These findings indicate that MFE01 uses 1-undecene emission for aerial communication, reporting for the first time this volatile organic compound as bacterial intraspecific communication signal.
PubMed: 37908545
DOI: 10.3389/fmicb.2023.1264801 -
Frontiers in Microbiology 2023Although the bacterial composition of boar ejaculate has been extensively studied, the bacterial composition of extended boar semen is often overlooked, despite the...
Although the bacterial composition of boar ejaculate has been extensively studied, the bacterial composition of extended boar semen is often overlooked, despite the potential risks these microorganisms may pose to the long-term preservation of extended boar semen at 15-17°C. In this study, we characterized the bacterial community composition of extended semen and discovered that spp. was the dominant flora. The dominant strains were further isolated and identified as a potential new species in the group and named strain, which had adverse effects on sperm quality and was better adapted to growth at 17°C. Antimicrobial susceptibility testing showed that the strain was resistant to all commonly used veterinary antibiotics. Whole-genome sequencing (WGS) and genome annotation revealed the large genetic structure and function [7,253,751 base pairs and 6,790 coding sequences (CDSs)]. Comparative genomic analysis with the closest type strains showed that the strain predicted more diversity of intrinsic and acquired resistance genes to multi-antimicrobial agents. Taken together, our study highlights a problem associated with the long-term storage of extended boar semen caused by a group strain with unique biological characteristics. It is essential to develop a new antibacterial solution for the long-term preservation of boar semen.
PubMed: 37869660
DOI: 10.3389/fmicb.2023.1279630 -
Scientific Reports Aug 2023The compound 2,4-diacetylphloroglucinol (DAPG) is a broad-spectrum antibiotic that is primarily produced by Pseudomonas spp. DAPG plays an important role in the...
The compound 2,4-diacetylphloroglucinol (DAPG) is a broad-spectrum antibiotic that is primarily produced by Pseudomonas spp. DAPG plays an important role in the biocontrol disease suppressing activity of Pseudomonas spp. In the current study, we report the discovery of the DAPG biosynthetic cluster in strains of Chromobacterium vaccinii isolated from Brazilian aquatic environments and the distribution of the biosynthetic cluster in the Chromobacterium genus. Phylogenetic analysis of the phlD protein suggests the biosynthetic cluster probably entered the genus of Chromobacterium after a horizontal gene transfer event with a member of the Pseudomonas fluorescens group. We were able to detect trace amounts of DAPG in wild type cultures and confirm the function of the cluster with heterologous expression in Escherichia coli. In addition, we identified and verified the presence of other secondary metabolites in these strains. We also confirmed the ability of C. vaccinii strains to produce bioactive pigment violacein and bioactive cyclic depsipeptide FR900359. Both compounds have been reported to have antimicrobial and insecticidal activities. These compounds suggest strains of C. vaccinii should be further explored for their potential as biocontrol agents.
Topics: Chromobacterium; Phylogeny; Anti-Bacterial Agents; Brazil; Escherichia coli; Pseudomonas
PubMed: 37653049
DOI: 10.1038/s41598-023-41277-0 -
Journal of Bacteriology Sep 2023Biofilm formation by the Gram-negative, Gammaproteobacteria relies on the repeats-in-toxin adhesins LapA and MapA in the cytoplasm, secretion of these adhesins through...
Biofilm formation by the Gram-negative, Gammaproteobacteria relies on the repeats-in-toxin adhesins LapA and MapA in the cytoplasm, secretion of these adhesins through their respective type 1 secretion systems, and retention at the cell surface. Published work has shown that retention of the adhesins occurs via a post-translational mechanism involving the cyclic-di-GMP receptor LapD and the protease LapG. However, little is known about the underlying mechanisms that regulate the level of these adhesins. Here, we demonstrate that the master regulator FleQ modulates biofilm formation by both transcriptionally and post-transcriptionally regulating LapA and MapA. We find that a Δ mutant has a biofilm formation defect compared to the wild-type (WT) strain, which is attributed in part to a decrease in LapA and MapA abundance in the cell, despite the Δ mutant having increased levels of and transcripts compared to the WT strain. Through transposon mutagenesis and subsequent genetic analysis, we found that overstimulation of the Gac/Rsm pathway partially rescues biofilm formation in the Δ mutant background. Collectively, these findings provide evidence that FleQ regulates biofilm formation by both transcriptionally regulating the expression of the genes and post-transcriptionally regulating the abundance of LapA and MapA, and that activation of the Gac/Rsm pathway can post-transcriptionally enhance biofilm formation by . IMPORTANCE Biofilm formation is a highly coordinated process that bacteria undergo to colonize a variety of surfaces. For , biofilm formation requires the production and localization of repeats-in-toxin adhesins to the cell surface. To date, little is known about the underlying mechanisms that regulate biofilm formation by . Here, we identify FleQ as a key regulator of biofilm formation that modulates both gene expression and abundance of LapA and MapA through both a transcriptional and post-transcriptional mechanism. We provide further evidence implicating activation of the Gac/Rsm system in FleQ-dependent regulation of biofilm formation. Together, our findings uncover evidence for a dual mechanism of transcriptional and post-transcriptional regulation of the LapA and MapA adhesins.
Topics: Pseudomonas fluorescens; Biofilms; Adhesins, Bacterial; Carrier Proteins; Bacterial Proteins; Gene Expression Regulation, Bacterial; Cyclic GMP
PubMed: 37655913
DOI: 10.1128/jb.00152-23 -
Anais Da Academia Brasileira de Ciencias 2023Pseudomonas fluorescens is known to have the ability to adhere and produce biofilm. The formation of biofilms is enhanced by cellular motility, particularly when...
Pseudomonas fluorescens is known to have the ability to adhere and produce biofilm. The formation of biofilms is enhanced by cellular motility, particularly when mediated by flagella. Biofilm formed on surfaces such as those used for food production act as points of contamination, releasing pathogenic or deteriorating microorganisms and compromising the quality of products. We assessed two strains of Pseudomonas fluorescens PL5.4 and PL7.1, sampled from raw, chilled, buffalo milk, which was obtained from a dairy farm. Twitching and swarming motility assays were performed, in addition to the biofilm production evaluations at a temperature of 7 °C. Regarding the motility assays, only the PL5.4 strain scored positive for the swarming assay. On microplates, both strains presented themselves as strong biofilm producers at 7 °C. The PL5.4 strain was also able to form biofilm on a stainless steel structure and maintain this structure for up to 72 hours at refrigeration. The Pseudomonas fluorescens PL5.4 isolate was identified on the basis of a 99% sequence identity with Pseudomonas fluorescens A506, a strain used as a biocontrol in agriculture. Biofilm-forming bacteria, when adapted to low temperatures, become a constant source of contamination, damaging the production, quality, safety and shelf-life of products.
Topics: Animals; Pseudomonas fluorescens; Milk; Biofilms; Temperature
PubMed: 37466543
DOI: 10.1590/0001-3765202320220982 -
Biofilm Dec 2023Biofilms are complex microbial communities embedded in extracellular matrix. Pathogens within the biofilm become more resistant to the antibiotics than planktonic...
Biofilms are complex microbial communities embedded in extracellular matrix. Pathogens within the biofilm become more resistant to the antibiotics than planktonic counterparts. Novel strategies are required to encounter biofilms. Exopolysaccharides are one of the major components of biofilm matrix and play a vital role in biofilm architecture. In previous studies, a glycosyl hydrolase, PslG, from was found to be able to inhibit biofilm formation by disintegrating exopolysaccharide in biofilms. Here, we investigate the potential spectrum of PslG homologous protein with anti-biofilm activity. One glycosyl hydrolase from , PslG, exhibits anti-biofilm activities and the key catalytic residues of PslG are conserved with those of PslG. PslG at concentrations as low as 50 nM efficiently inhibits the biofilm formation of and disassemble its preformed biofilm. Furthermore, PslG exhibits anti-biofilm activity on a series of , including and pv. . PslG stays active under various temperatures. Our findings suggest that glycosyl hydrolase PslG has potential to be a broad spectrum inhibitor on biofilm formation of a wide range of .
PubMed: 37928620
DOI: 10.1016/j.bioflm.2023.100155 -
The ISME Journal Oct 2023Proteobacteria primarily utilize acyl-homoserine lactones (AHLs) as quorum-sensing signals for intra-/interspecies communication to control pathogen infections....
Proteobacteria primarily utilize acyl-homoserine lactones (AHLs) as quorum-sensing signals for intra-/interspecies communication to control pathogen infections. Enzymatic degradation of AHL represents the major quorum-quenching mechanism that has been developed as a promising approach to prevent bacterial infections. Here we identified a novel quorum-quenching mechanism revealed by an effector of the type IVA secretion system (T4ASS) in bacterial interspecies competition. We found that the soil antifungal bacterium Lysobacter enzymogenes OH11 (OH11) could use T4ASS to deliver the effector protein Le1288 into the cytoplasm of another soil microbiome bacterium Pseudomonas fluorescens 2P24 (2P24). Le1288 did not degrade AHL, whereas its delivery to strain 2P24 significantly impaired AHL production through binding to the AHL synthase PcoI. Therefore, we defined Le1288 as LqqE1 (Lysobacter quorum-quenching effector 1). Formation of the LqqE1-PcoI complex enabled LqqE1 to block the ability of PcoI to recognize/bind S-adenosy-L-methionine, a substrate required for AHL synthesis. This LqqE1-triggered interspecies quorum-quenching in bacteria seemed to be of key ecological significance, as it conferred strain OH11 a better competitive advantage in killing strain 2P24 via cell-to-cell contact. This novel quorum-quenching also appeared to be adopted by other T4ASS-production bacteria. Our findings suggest a novel quorum-quenching that occurred naturally in bacterial interspecies interactions within the soil microbiome by effector translocation. Finally, we presented two case studies showing the application potential of LqqE1 to block AHL signaling in the human pathogen Pseudomonas aeruginosa and the plant pathogen Ralstonia solanacearum.
Topics: Humans; Quorum Sensing; Bacterial Proteins; Pseudomonas; Pseudomonas aeruginosa; Acyl-Butyrolactones; Pseudomonas fluorescens
PubMed: 37340074
DOI: 10.1038/s41396-023-01457-2 -
Journal of Photochemistry and... Jul 2023As a new kind of modern military biological weapon, bacterial agents pose a serious threat to the public health security of human beings. Existing bacterial...
As a new kind of modern military biological weapon, bacterial agents pose a serious threat to the public health security of human beings. Existing bacterial identification requires manual sampling and testing, which is time-consuming, and may also introduce secondary contamination or radioactive hazards during decontamination. In this paper, a non-contact, nondestructive and "green" bacterial identification and decontamination technology based on laser-induced breakdown spectroscopy (LIBS) is proposed. The principal component analysis (PCA) combined with support vector machine (SVM) based on radial basis kernel function is used to establish the classification model of bacteria, and the two-dimensional decontamination test of bacteria is carried out using laser-induced low-temperature plasma combined with a vibration mirror. The experimental results show that the average identification rate of the seven types of bacteria, including Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, Bacillus megatherium, Pseudomonas aeruginosa, Bacillus thuringiensis and Enterococcus faecalis reaches 98.93%, and the corresponding true positive rate, precision, recall and F1-score reaches 0.9714, 0.9718, 0.9714 and 0.9716, respectively. The optimal decontamination parameters are laser defocusing amount of -50 mm, laser repetition rate of 15-20 kHz, scanning speed of 150 mm/s and number of scans of 10. In this way, the decontamination speed can reach 25.6 mm/min, and the inactivation rates for both Escherichia coli and Bacillus subtilis are higher than 98%. In addition, it is confirmed that the inactivation rate of plasma is 4 times higher than that of thermal ablation, meaning that the decontamination ability of LIBS mainly relies on the plasma rather than the thermal ablation effect. The new non-contact bacterial identification and decontamination technology does not require sample pretreatment, and can quickly identify bacteria in situ and decontaminate the surfaces of precision instruments, sensitive materials, etc., which has potential application value in modern military, medical and public health fields.
Topics: Humans; Decontamination; Bacillus subtilis; Spectrum Analysis; Lasers; Escherichia coli
PubMed: 37201319
DOI: 10.1016/j.jphotobiol.2023.112719 -
Bioinformation 2023Increased amounts of toxicants may cause sever health issues in humans as well as in aquatic life. Scientists are developing new technologies to combat these problems....
Increased amounts of toxicants may cause sever health issues in humans as well as in aquatic life. Scientists are developing new technologies to combat these problems. Biological methods of detoxification are always beneficial for the environment. Pseudomonas fluorescens is known for its detoxification capacity. In this study stains were isolated from different locations of the Ha'il region, Saudia Arabia. The microbial strain AM-1 displayed resistance to heavy metals (Cr6+, Ni2+, Cd2+, Pb2+) and pesticides (BHC, 2,4-D, Mancozeb) at pollutant levels typical of highly contaminated areas. Additionally, AM-1 exhibited substantial detoxification potential, reducing toxicity by 40.67% for heavy metals and 47.4% for pesticides at 3x concentrations. These findings suggest that the AM-1 strain supports environmental remediation and pollution mitigation. Atomic absorption spectrometry (AAS) results exhibited bioremediation efficiency for metals Cr, Ni, and Pb using immobilized cells of AM-1 isolate, estimated to be 60.57%, 68.4%, and 53.93% respectively. These findings show that AM-1 strain has a potential role in bioremediation of water pollutants and may have future implications in wastewater treatment.
PubMed: 37928494
DOI: 10.6026/97320630019901