Fems Microbiology Letters最新文献

SMC (Structural Maintenance of Chromosomes) ATPase proteins are integral components of complexes bearing the same name, crucial for the spatial organization of DNA across diverse life forms, spanning bacteria, archaea, and eukaryotes. It is proposed that in bacteria, SMC complexes facilitate DNA compaction through loop extrusion and aid in the segregation of daughter nucleoids. In this paper, the properties of the SMC ATPase protein from Ureaplasma parvum were investigated by using a spectrum of methods, including conventional biochemical methods as well as advanced single-molecule techniques. Our findings reveal distinctive properties of this protein compared to its extensively studied homologue from Bacillus subtilis. Notably, our results suggest that U. parvum Smc ATPase facilitates DNA compaction even in the absence of ATP.

Bacteria that ferment amino acids to ammonia can be categorized as generalists or specialist hyper-ammonia-producing bacteria. In the rumens of ruminant animals, most of the ammonia produced is eventually excreted as urea in urine. This process can be controlled with off-label use of antibiotics, but the practice can lead to antibiotic resistance; therefore, discovery of antibiotic alternatives is pertinent. Plant-derived phenolic compounds have demonstrated antimicrobial efficacy for such purposes. This study investigated the antimicrobial and metabolic suppressive potential of six phenolic compounds on five amino acid fermenting bacteria: Clostridium sporogenes MD1, C. aminophilum F, Acetoanaerobium sticklandii SR, Peptostreptococcus sp. BG1, and Prevotella bryantii B14. Inhibitory action of the compounds was determined using a 10% v/v serial dilution method in basal media. Carvacrol (1 mM), thymol (1 mM), and eugenol (10 mM) demonstrated the greatest antimicrobial potential, where carvacrol and eugenol inhibited growth of all five species and thymol four species except BG1. The cinnamic acids (trans and hydro) demonstrated variable activity against all organisms. Suppression of metabolic activity was determined via colorimetric assay quantifying ammonia in washed stationary phase culture supernatant after 24 h of metabolism on fresh substrate. Carvacrol and eugenol yielded the greatest reduction of ammonia by all organisms except B14, which produced no ammonia under the growth conditions. Thymol greatly reduced ammonia production of four organisms except F. These data demonstrate that eugenol, carvacrol, and thymol may be worthy antimicrobial candidates for the control of ammonia-producing organisms.

In this study designed to isolate lactic acid bacteria (LAB) with bacteriocin production potential, white cheese samples were collected from different provinces of Turkey and isolation was carried out. A series of experiments were carried out for the main purpose and the actual bacteriocin producers were identified by detecting the genes encoding this bacteriocin. The experiments carried out in this direction were initially carried out with 20 isolates and as a result of various experiments, the number of isolates was reduced to 8 and the study was continued with 8 isolates. In order to determine that the eight isolates identified as a result of a phenotypic and biochemical characterization study were true bacteriocin-producing strains, their antibacterial activity was investigated and then the presence of bacteriocin genes was examined by specific polymerase chain reaction (PCR) using gene-specific primers. As a result, MS16 coded Lactiplantibacillus plantarum OR922652 was found to have strong antibacterial activity against Escherichia coli, Klebsiella pneumonia, Yersinia enterocolitica, Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus; the isolate was susceptible to clinically important antibiotics (ciprofloxacin, gentamicin, penicillin G, ampicillin, chloramphenicol, and vancomycin) and resistant to erythromycin, had no hemolytic activity and possessed plnA and plnD genes encoding bacteriocin production. In conclusion, the MS16 coded L. plantarum isolate has emerged as a promising strain that can be used especially in the health field and in the food industry related to LAB.

Pseudomonas aeruginosa has increasing clinical relevance and commonly occupies the cystic fibrosis (CF) airways. Its ability to colonize and persist in diverse niches is attributed to its large accessory genome, where prophages represent a common feature and may contribute to its fitness and persistence. We focused on the CF airways niche and used 197 longitudinal isolates from 12 patients persistently infected by P. aeruginosa. We computationally predicted intact prophages for each longitudinal group and scored their long-term persistence. We then confirmed prophage inducibility and mapped their location in the host chromosome with lysate sequencing. Using comparative genomics, we evaluated prophage genomic diversity, long-term persistence, and level of genomic maintenance. Our findings support previous findings that most P. aeruginosa genomes harbour prophages some of which can self-induce, and that a common CF-treating antibiotic, ciprofloxacin, can induce prophages. Induced prophage genomes displayed high diversity and even genomic novelty. Finally, all induced prophages persisted long-term with their genomes avoiding gene loss and degradation over 4 years of host replication in the stressful CF airways niche. This and our detection of phage genes, which contribute to host competitiveness and adaptation, lends support to our hypothesis that the vast majority of prophages detected as intact and inducible in this study facilitated their host fitness and persistence.

Crystal-violet staining, microscopy with image analysis, and quantitative PCR (qPCR) were compared to examine biofilm dynamics. Biofilms of 30 polycultures comprising 15 bacterial species were monitored for 14 days. Collectively, qPCR (representing population) revealed a different growth pattern compared to staining (biomass) and microscopy (colonization): biomass and colonization gradually increased over time, whereas population increased rapidly for the first seven days and leveled off. Temporal forms were categorized into two growth patterns: continuous increase (CI) and non-continuous increase. Staining and microscopy showed similar odds of detecting the CI pattern (27 and 23 polycultures, respectively) across polycultures, greater than that of qPCR (14 polycultures) (P < 0.05). All three methods revealed the identical patterns for 13 polycultures. Staining with microscopy, staining with qPCR, and microscopy with qPCR found the same patterns in 22, 15, and 19 polycultures, respectively. Additionally, staining was quantitatively agreed with microscopy (P < 0.05; R2 > 0.50), whereas neither staining nor microscopy strongly agreed with qPCR (P < 0.05; R2 ≤ 0.22). Collectively, staining was more compatible with microscopy than qPCR in characterizing biofilm dynamics and quantifying biofilms owing to the difference between population growth and biofilm expansion. The concurrent use of qPCR with biomass estimations allows for accurate and comprehensive biofilm quantification.

SARS-CoV-2 infection is an acute respiratory distress syndrome associated with immune dysfunction, causing coronavirus disease 2019 (COVID-19) disease. The use of immunosuppressive drugs in its treatment increases the risk of opportunistic infections. In particular, opportunistic fungal infections have been described in initially non-immunocompromised patients with severe COVID-19 disease. Among them, rare cases of cryptococcosis have been described. Here we present the first two French cases of non-HIV non-transplant patients who developed disseminated Cryptococcus neoformans fungal infection in the setting of severe COVID-19 disease. Blood cultures appear to be an interesting diagnostic tool for post-COVID-19 cryptococcosis, which is an often fatal complication.

Online social networks have revolutionized scientific communication, making platforms like Instagram indispensable for sharing complex topics, including mycology. This study evaluated three key factors in assessing the impact of social media on scientific dissemination: follower profiles, reach, and engagement levels. We used the professional Instagram account @microbioworld as a case study. Account performance data were collected over a 90-day period (12 August-9 November 2024). Post performance was evaluated using data from selected posts published between 11 January and 11 November 2024. By the end of our sampling period, the account reached a total of 45 959 followers, with the majority aged 25-34 years (44.8%). It reached 108 631 unique accounts, with 22.4% being followers and 77.6% non-followers, generating 236 860 impressions and 15 750 interactions. Likes accounted for 83.3% of engagement. Posts featuring microorganism cultures achieved the highest engagement and reach. Sentiment analysis, using Bing and AFINN lexicons, revealed that over 89% of sentiments expressed in comments were positive. These findings demonstrate how Instagram can disseminate microbial content, foster positive perceptions of microorganisms, and emphasize their ecological importance, encouraging audience involvement with mycology and microbiology.

Bacterial transcription terminator, Rho is an RNA (Ribonucleic Acid)-dependent ATPase that terminates transcription. Several structures of pretermination complexes of the Rho-transcription elongation complex (EC) revealed a static picture of components of the EC that come close to the nascent RNA-bound Rho, where many of the residues of EC reside ≤10 Å from the Rho residues. However, the in vitro-formed Rho-EC complexes do not reveal the in vivo Rho-EC dynamic interaction patterns during the termination process. Here we report synthetic defect analyses of various combinations of the mutations in RNAP β, β' and ω-subunits, NusA, NusG, and Rho proteins to delineate the functional network of this process. Several mutations in the β-flap and β'-Zn-finger and -Clamp helices domains of RNAP are synthetically defective in the presence of Rho mutants indicating functional involvement of these domains. Mutations in the NusA RNA-binding domains were synthetically defective with the Rho mutants suggesting its involvement. Our genetic analyses also revealed functional antagonisms between the ω-subunit of RNAP and the NusG-CTD (c-terminal domain) during termination. We concluded that the regions surrounding the RNA exit channel, the RNA-binding domains of NusA, the RNAP ω-subunit, and NusG-CTD constitute a functional network with Rho just before the onset of in vivo Rho-dependent termination.

Hemolytic proteins are a major group of virulence factors in pathogenic Aeromonas hydrophila. Six genes encoding presumable hemolytic proteins were revealed from the genome of virulent A. hydrophila (vAh) that caused severe disease in channel catfish. The aim of this study was to assess the contribution of these hemolytic proteins to the virulence of this bacterium. Genes coding for following six proteins were investigated: aerolysin (Arl), 21-kDa hemolysin (Hly1), thermostable hemolysin (Hly2), phospholipase/lecithinase-related hemolysin (Hly3), membrane-associated hemolysin III (Hly4), and cytolysin-associated hemolysin (Hly5). Individual genes were deleted from the bacterium using CRISPR-Cas9 mediated methods. Assessment showed that deletion of Arl gene (Δarl) completely abolished hemolytic activity of this mutant while Δhly1-Δhly5 mutants had the same activity as the wild vAh. Extracellular proteins (ECPs) of the Δarl mutant caused significantly (p < 0.01) less cell death in vitro with viability increased by approximately 20%, compared to the wild vAh. ECPs of mutants Δhly1-Δhly5 remained the same cell toxicity as the wild vAh. A second deletion of hly5 from the Δarl mutant further lowered the cell toxicity of the ECP of the mutant (Δarl + Δhly5). Assays in vivo showed that both Δarl and Δhly5 mutants caused less fish mortality with reduction of 57% and 16%, respectively, compared to the wild vAh; the Δarl + Δhly5 mutant caused the least mortality with approximately 87% of reduction; and other mutants had the same virulence as the wild vAh. Analyses of SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Western blotting evidently indicate that both Arl and Hly5 proteins formed hexamer-like stable structures post secretion from the bacterium. Arl and Hly5 apparently had synergistic action in cytotoxicity and causing disease and were the major virulence factors among the six hemolytic proteins analyzed in this study.

Stenotrophomonas maltophilia is an emerging global opportunistic pathogen that causes nosocomial infections. We demonstrated that the superoxide stress-sensing transcriptional regulator SoxR directly modulated the expression of an operon encompassing sodA1 (encoding manganese-containing superoxide dismutase) and fre (encoding putative flavin reductase) by directly binding to the operator site, which was located between the -35 and -10 motifs of the sodA1 promoter. It is known that upon exposure to the superoxide generators/redox-cycling drugs, the SoxR, which is bound to the operator site, became oxidized. This oxidation causes a conformational change of SoxR to an active form, enabling the upregulation of sodA1-fre gene expression. A ΔsodA1 was constructed, and the mutant showed enhanced sensitivity to the redox-cycling drugs, including menadione, plumbagin, and methyl viologen (paraquat), relative to its parental strain K279a. Thus, sodA1 may play a role in the survival of S. maltophilia under superoxide stress during either its saprophyte stage (e.g. exposure to redox-cycling drugs) or host-pathogen interactions.