Phase Variation of LPS and Capsule Is Responsible for Stochastic Biofilm Formation in Francisella tularensis

Biofilms have been established as an important lifestyle for bacteria in nature as these structured communities often enable survivability and persistence in a multitude of environments. Francisella tularensis is a facultative intracellular Gram-negative bacterium found throughout much of the northern hemisphere. However, biofilm formation remains understudied and poorly understood in F. tularensis as non-substantial biofilms are typically observed in vitro by the clinically relevant subspecies F. tularensis subsp. tularensis and F. tularensis subsp. holarctica (Type A and B, respectively). Herein, we report conditions under which robust biofilm development was observed in a stochastic, but reproducible manner in Type A and B isolates. The frequency at which biofilm was observed increased temporally and appeared switch-like as progeny from the initial biofilm quickly formed biofilm in a predictable manner regardless of time or propagation with fresh media. The Type B isolates used for this study were found to more readily switch on biofilm formation than Type A isolates. Additionally, pH was found to function as an environmental checkpoint for biofilm initiation independently of the heritable cellular switch. Multiple colony morphologies were observed in biofilm positive cultures leading to the identification of a particular subset of grey variants that constitutively produce biofilm. Further, we found that constitutive biofilm forming isolates delay the onset of a viable non-culturable state. In this study, we demonstrate that a robust biofilm can be developed by clinically relevant F. tularensis isolates, provide a mechanism for biofilm initiation and examine the potential role of biofilm formation.

Bacteroidales species in the human gut are a reservoir of antibiotic resistance genes regulated by invertible promoters

Antibiotic-resistance genes (ARGs) regulated by invertible promoters can mitigate the fitness cost of maintaining ARGs in the absence of antibiotics and could potentially prolong the persistence of ARGs in bacterial populations. However, the origin, prevalence, and distribution of these ARGs regulated by invertible promoters remains poorly understood. Here, we sought to assess the threat posed by ARGs regulated by invertible promoters by systematically searching for ARGs regulated by invertible promoters in the human gut microbiome and examining their origin, prevalence, and distribution. Through metagenomic assembly of 2227 human gut metagenomes and genomic analysis of the Unified Human Gastrointestinal Genome (UHGG) collection, we identified ARGs regulated by invertible promoters and categorized them into three classes based on the invertase-regulating phase variation. In the human gut microbiome, ARGs regulated by invertible promoters are exclusively found in Bacteroidales species. Through genomic analysis, we observed that ARGs regulated by invertible promoters have convergently originated from ARG insertions into glycan-synthesis loci that were regulated by invertible promoters at least three times. Moreover, all three classes of invertible promoters regulating ARGs are located within integrative conjugative elements (ICEs). Therefore, horizontal transfer via ICEs could explain the wide taxonomic distribution of ARGs regulated by invertible promoters. Overall, these findings reveal that glycan-synthesis loci regulated by invertible promoters in Bacteroidales species are an important hotspot for the emergence of clinically-relevant ARGs regulated by invertible promoters.

Comparative Genomics Revealed Genus Specific Encoding of Amino Acids by Tri-Nucleotide SSRs in Human Pathogenic Streptococcus and Staphylococus Bacteria

Pathogenic bacteria use phase variation of surface molecules and other characteristics as a significant adaptation mechanism. Repetitive sequences made up of numerous identical repeat units can be found in many phase variable genes. Here, we investigated the frequency and distribution of long-SSRs in 15 human pathogenic Staphylococcus, Streptococcus, and Enterococcus bacteria. Long-SSRs were found to be distributed differently in the genic and intergenic sequences. In the genic sequences, 61.3 SSRs were discovered on average, while 16.2 SSRs were found in the intergenic regions. Staphylococci exhibited the highest frequency of SSRs, followed by Enterococcus, and Streptococci had the lowest frequency of SSRs. Higher A+T content was found to be the best predictor of long-SSR in these human pathogens. Tetranucleotide repeats predominated in intergenic regions, while trinucleotide repeats predominated in genic regions. In human pathogenic Streptococcus and Staphylococcus bacteria, genus-specific encoding of amino acids by tri-nucleotide SSRs was observed. A genetic relationship between these human pathogenic bacteria was derived based on the presence of SSRs in the housekeeping genes and compared to the phylogeny generated based on the 16S ribosomal RNA gene.

Theoretical study on the phase sensitivity of condenser type acoustic transducers at low frequencies

The phase sensitivity of the condenser type acoustic transducers at low frequencies is crucial for locating large-scale natural and manmade activities, but is now commonly calibrated based on comparison methods. Although the primary method, which traces its sensitivity back to the international standard unit is few studied. Recently, the explicit sensitivity models of the condenser type acoustic transducers based on the laser-pistonphone technique are built, and can be used to study the phase responses of acoustic transducers at infrasonic frequencies. So that, in this paper, the phase sensitivities of acoustic transducers when its rear vent connected to the calibrating sound field or outside atmosphere are studied in detail. Secondly, time domain analysis of generated sound pressures by displacement excitation are derived to reveal the mechanism of phase variation. Calculations show two distinct sensitivities with 90° phase lead and −10° phase lag limits for vent in field and vent out field calibrations, which are dominated by the pressure leakage and heat conduction effects at infrasonic frequencies.

What Flips the Switch? Signals and Stress Regulating Extraintestinal Pathogenic Escherichia coli Type 1 Fimbriae (Pili)

Pathogens are exposed to a multitude of harmful conditions imposed by the environment of the host. Bacterial responses against these stresses are pivotal for successful host colonization and pathogenesis. In the case of many E. coli strains, type 1 fimbriae (pili) are an important colonization factor that can contribute to diseases such as urinary tract infections and neonatal meningitis. Production of type 1 fimbriae in E. coli is dependent on an invertible promoter element, fimS, which serves as a phase variation switch determining whether or not a bacterial cell will produce type 1 fimbriae. In this review, we present aspects of signaling and stress involved in mediating regulation of type 1 fimbriae in extraintestinal E. coli; in particular, how certain regulatory mechanisms, some of which are linked to stress response, can influence production of fimbriae and influence bacterial colonization and infection. We suggest that regulation of type 1 fimbriae is potentially linked to environmental stress responses, providing a perspective for how environmental cues in the host and bacterial stress response during infection both play an important role in regulating extraintestinal pathogenic E. coli colonization and virulence.

Analyses of wave-phase variation of Reynolds shear stress underneath surface wave using streamline coordinates

The dependence of Reynolds shear stress on wave phase is investigated for initially isotropic turbulence distorted by a progressive surface wave through direct numerical simulations. A wave-following streamline coordinate frame is used to analyse the turbulence dynamics such that the information of the varying direction of wave orbital motions is embedded into the coordinate system, which helps capture the effect of flow curvature on the turbulence and quantify momentum exchange between the near-surface and deep regions in the wavy domain accurately. It is found that the Reynolds shear stress is enhanced under the backward slope of the wave and can be scaled by the wave steepness and the streamwise velocity fluctuations. Analyses of the budget of Reynolds shear stress indicate that such a variation with the wave phase is caused by the variation in the production of the Reynolds shear stress and the effect of pressure fluctuations. Further investigation shows that the production of the Reynolds shear stress is closely associated with the wave surface curvature. A model that includes a correction term for the curvature effect for the pressure–strain correlation term is examined and is found to agree reasonably well with the simulation result. The correction term is found to make an appreciable contribution to the model, further supporting our finding that the wave curvature plays an important role in the turbulence dynamics near the surface.

Global Sensitivity Analysis for CERES-Rice Model under Different Cultivars and Specific-Stage Variations of Climate Parameters

Global sensitivity analysis (SA) has become an efficient way to identify the most influential parameters on model results. However, the effects of cultivar variation and specific-stage variations of climate conditions on model outputs still remain unclear. In this study, 30 indica hybrid rice cultivars were simulated in the CERES-Rice model; then the Sobol’ method was used to perform a global SA on 16 investigated parameters for three model outputs (anthesis day, maturity day, and yield). In addition, we also compared the differences in the sensitivity results under four specific-stage variations (vegetative phase, panicle-formation phase, ripening phase, and the whole growth season) of climate conditions. The results indicated that (1) parameter Tavg, G4, and P2O are the most influential parameters for all model outputs across cultivars during the whole growth season; (2) under the vegetative-phase variation of climate parameters; the variability of model outputs is mainly controlled by parameter P2O and Tavg; (3) under the panicle-formation-phase or ripening-phase variation of climate parameters, parameter P2O was the dominant variable for all model outputs; (4) parameter PORM had a considerable effect (the total sensitivity index, STi; STi>0.05) on yield regardless of the various specific-stage variations of the climate parameters. Findings obtained from this study will contribute to understanding the comprehensive effects of crop parameters on model outputs under different cultivars and specific-stage variations of climate conditions.