Swarming Motility Research Articles

Biofilm Control by Active Topography with Mucin Coating

AbstractIn the innate immune system of mammalians, beating cilia of epithelial cells and the attached mucin proteins prevent the colonization of microbial pathogens. Abiotic biomaterials of medical implants lack such protection and thus are susceptible to microbial colonization, leading to biofilm formation and persistent infections with high‐level antibiotic tolerance. To address this challenge, the team further develops its new strategy of biofilm control by magnetically driven oscillation of micron‐sized pillars on biomaterials. This study is based on a bioinspired design by covalently coating the pillars with mucin, a glycoprotein found ubiquitously in mammalian innate immune systems. The results show that mucin coating significantly enhances the antifouling effects of active topography in both the inhibition of Pseudomonas aeruginosa biofilm formation and removal of its mature biofilms. Analysis using scanning electron microscopy (SEM) reveals that mucin coating inhibits bacterial attachment near the pillar base, the area protected from the direct force of beating pillars. In addition, mucin coating enhances the twitching motility of P. aeruginosa but represses its swarming motility and the synthesis of cyclic di‐GMP. These effects further contribute to the antifouling activities. Overall, these findings demonstrate the feasibility of engineering bioinspired antifouling materials for safer medical devices.

Physiological adaptations of clinical vs. indoor environmental strains of Pseudomonas aeruginosa in a hospital setting.

Physiological adaptations of clinical vs. indoor environmental strains of Pseudomonas aeruginosa in a hospital setting.

Correlation Analysis Between Multi-Drug Resistance Phenotype and Virulence Factor Expression of Clinical Pseudomonas aeruginosa.

Pseudomonas aeruginosa (PA), as a common pathogen of nosocomial infections, has been experiencing an increasing rate of drug resistance with the widespread use and abuse of antimicrobial drugs. High-drug-resistance and high-virulence phenotypes are two distinctive features of the strong pathogenicity of multi-drug-resistant PA. Exploring the characterization of virulence factor expression and its relationship with the multi-drug resistance phenotype is essential to reduce the further development of resistance as well as a high standard of infection prevention and control. A total of 50 PA isolated from clinical practice were collected. The Kirby-Bauer test was used for drug-sensitive screening, and the results showed that 16 strains were resistant and 16 strains were sensitive. The drug resistance rate of multi-drug-resistant PA against cefepime, cefazolin, ampicillin, and imipenem was up to 100%. The multi-drug-resistant groups were superior in producing pyocyanin and forming biofilm to the sensitive groups. The distribution of isolates with different swarming motility capacities and elastase levels did not show pronounced differences among the multi-drug-resistant and sensitive groups. In addition, biofilm formation was moderately associated with imipenem resistance. Among the strains with strong virulence factor expression, the gene bands showed little difference, suggesting that the gene is highly homologous. The virulence factor matrix analysis showed that there were different degrees of correlation among the 4 virulence factors. The correlation between multidrug-resistant PA and virulence factor expression is complex. PA, which were good at producing pyocyain and forming biofilm, were highly resistant to cephalosporins, beta-lactams and carbepenems; hence, such drugs are not proper for anti-infective treatment in clinics.

Vibrio parahaemolyticus Down-Regulates the Intracellular c-di-GMP Level to Promote Swarming Motility by Sensing Surface.

Vibrio parahaemolyticus, a significant food-borne pathogen that causes economic and public health problems worldwide, can produce two types of flagella, the single polar flagellum responsible for swimming in a liquid environment and the lateral flagella (laf) that enable the bacteria to swarm on the tops of solid surfaces. The polar flagellar genes are expressed either in liquid or on a surface, however, laf genes would only be activated by surface sensing. In this study, the molecular mechanism of surface sensing activating laf gene expression in V. parahaemolyticus was investigated. We found that the c-di-GMP concentration for liquid-grown cells was higher than the concentration for surface-grown cells and laf gene expression could be activated without touching surface if the intracellular level of c-di-GMP was decreased by overexpressing the scrABC operon in the wild-type strain. Surface sensing inhibits the transcription of those c-di-GMP-metabolizing enzymes which could negatively regulate swarming, and enhances those which could positively regulate swarming. Surface sensing also enhances the activity of quorum sensing (QS) of V. parahaemolyticus which plays an important role in regulating the transcription of components of the c-di-GMP network. Combined, the data indicate that V. parahaemolyticus enhances QS by surface sensing to down-regulate the intracellular level of c-di-GMP which results in producing the lateral flagella and swarming on a surface. What we found in this study suggests an important signal transduction pathway of regulating swarming motility by surface sensing in V. parahaemolyticus.

LafK contributes the regulation of swarming motility of Pseudomonas plecoglossicida and bacterial-host interaction

lafK contributes the regulation of swarming motility of Pseudomonas plecoglossicida and bacterial-host interaction

Evaluation of HPLC Profile, Antioxidant, Quorum Sensing, Biofilm, Swarming Motility, and Enzyme Inhibition Activities of Conventional and Green Extracts of Salvia triloba.

The current study aims to prepare a green extract using a new method in addition to conventional extraction methods including; methanolic and ultrasonic extraction of Salvia triloba, to compare their phenolic composition utilizing high-performance liquid chromatograph equipped with a diode array detector (HPLC-DAD), anti-bacterial, anti-oxidant, and enzyme inhibition activities. The results of HPLC-DAD analysis showed that Rosmarinic acid was found the highest amount in the methanolic extract followed by ultrasonic and green extracts as 169.7 ± 0.51, 135.1 ± 0.40, and 28.58 ± 0.46 μg/g respectively. The Trans-cinnamic acid (4.40 ± 0.09 μg/g) was found exclusively in ultrasonic extract. For bioactivities, the green extract exhibited the highest biofilm inhibition against Enterococcus faecalis compared to other extracts, while the methanolic extract outperformed both ultrasonic-assisted and green extract against Staphylococcus aureus and Escherichia coli strains at minimum inhibitory concentration. The methanolic and green extract exhibited considerable quorum sensing inhibition against Chromobacterium violaceum CV026, while no activity was recorded from ultrasonic-assisted extract. The methanolic and ultrasonic-assisted extracts of S. triloba recorded moderate butyrylcholinesterase inhibition; each extract demonstrated limited inhibitory effects on the urease enzyme. Similarly, each extract of S. triloba demonstrated significant antioxidant activity, with the highest activity exhibited by methanolic extract as β-carotene-linoleic acid assay (IC50 = 10.29 ± 0.36 μg/mL), DPPH• assay (IC50 = 27.77 ± 0.55 μg/mL), ABTS•+ assay (IC50 = 15.49 ± 0.95 μg/mL), metal chelating assay (IC50 = 57.80 ± 0.95 μg/mL), and CUPRAC (assay A 0.50 = 32.54 ± 0.84 μg/mL). Furthermore, the methanolic extract exhibited antioxidant activity better than α-tocopherol (Standard used). The current study demonstrated the potential of green solvent(s) as eco-friendly alternative for extractin phenolic compounds from S. triloba and evaluated their biological activities for the first time.

Effect of novel non-coding small RNA111 of Pseudomonas aeruginosa on bacterial virulence

Effect of novel non-coding small RNA111 of Pseudomonas aeruginosa on bacterial virulence

Inhibition of Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms by grape and rice agroindustrial residues

Inhibition of Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms by grape and rice agroindustrial residues

Effect of Lactobacillus supernatant on swarming-related gene expression in Proteus mirabilis isolated from urinary tract infections

Proteus mirabilis isolates have been intensively researched for their capacity to cause urinary tract infections (UTIs) and their swarming motility, although little is known about this phenomenon. Probiotic Lactobacillus species, which are beneficial bacteria, are being studied worldwide as therapeutic and preventative agents against bacterial infections. This study investigated Lactobacillus supernatants as a potential new treatment against Proteus mirabilis. In addition to testing their antimicrobial and anti-swarming activities, the research also aimed to understand the genetic mechanisms behind the observed phenotypic changes. Methods. A total of 150 urine specimens were collected from UTI patients at various hospitals in Baghdad. Direct culture was performed by streaking the specimens on differential media. RNA was extracted and purified from the bacterial isolates, and then reverse transcription and quantitative PCR were used to evaluate swarming-related gene expression. Gene expression was assessed relative to a reference gene to reveal how probiotics regulate swarming behavior at the genetic level. Gene expression patterns varied, indicating complex genomic responses to Lactobacillus exposure. Results. UTIs affected 50 males (33.33%) and 100 females (66.66%) of various ages. Proteus mirabilis was identified in 30 (20%) of the 150 samples. Resistance was observed in 25 (83.33%) isolates for azithromycin and amoxicillin/clavulanic acid, and in 22 (73.33%) isolates for meropenem. Real-time PCR showed significant alterations in the expression of four swarming-related genes (rsbA, umoD, ZapA, and FliL). The rsbA gene showed a notable increase in expression, while another sample displayed a decrease. The umoD gene exhibited the largest change, with expression doubling in some cases. ZapA showed the greatest increase, nearly tripling in expression in one sample. FliL expression also rose in multiple isolates. Swarming activity was positively correlated with gene expression levels for rsbA (r = 0.8, p = 0.009), umoD (r = 0.635, p = 0.045), ZapA (r = 0.942, p = 0.001), and FliL (r = 0.894, p = 0.001). Conclusions. The study reveals a complex gene network regulating the swarming motility of Proteus mirabilis. It suggests that Lactobacillus acidophilus supernatants can modify gene expression and bacterial motility, potentially aiding in the treatment of UTIs.

Surfactins and Iturins Produced by Bacillus velezensis Jt84 Strain Synergistically Contribute to the Biocontrol of Rice False Smut

Rice false smut, caused by the plant pathogenic fungus Ustilaginoidea virens, is widespread in rice-growing regions globally, severely compromising rice quality and production. Employing Bacillus spp. to control rice false smut represents an effective and environmentally sustainable strategy for disease management. The lipopeptides produced by Bacillus velezensis Jt84 demonstrated robust inhibitory effects against U. virens, resulting in abnormal mycelial morphology and spore germination. Iturins were identified as essential for the antifungal activity against U. virens, as confirmed by mutagenesis experiments that suppressed iturin biosynthesis. The surfactin-deficient mutant exhibited inhibitory effects against U. virens comparable to the wild-type, indicating that the absence of surfactins did not diminish its antifungal activity. Both the Jt84∆srf and Jt84∆itu mutants displayed reduced biofilm formation capabilities compared to the wild-type, with the Jt84∆srf mutant being particularly impaired and unable to form a complete biofilm. Regarding swarming motility, the ∆srf mutant exhibited a significant reduction compared to the wild-type, whereas the Jt84∆itu mutant showed a modest increase. Colonization experiments revealed that the Jt84∆srf mutant strain had significantly lower colonization on rice leaf surfaces than the wild-type strain, highlighting the critical role of surfactins in the colonization of B. velezensis Jt84 on rice leaves. In conclusion, our research demonstrated that surfactins and iturins have distinct functionalities and act synergistically to contribute to the biocontrol of rice false smut in B. velezensis Jt84. This synergy is achieved through their potent antifungal effects, biofilm formation, and successful colonization.

Impact of Quorum Sensing on the Virulence and Survival Traits of Burkholderia plantarii.

Quorum sensing (QS) is a mechanism by which bacteria detect and respond to cell density, regulating collective behaviors. Burkholderia plantarii, the causal agent of rice seedling blight, employs the LuxIR-type QS system, common among Gram-negative bacteria, where LuxI-type synthase produces QS signals recognized by LuxR-type regulators to control gene expression. This study aimed to elucidate the QS mechanism in B. plantarii KACC18965. Through whole-genome analysis and autoinducer assays, the plaI gene, responsible for QS signal production, was identified. Motility assays confirmed that C8-homoserine lactone (C8-HSL) serves as the QS signal. Physiological experiments revealed that the QS-defective mutant exhibited reduced virulence, impaired swarming motility, and delayed biofilm formation compared to the wild type. Additionally, the QS mutant demonstrated weakened antibacterial activity against Escherichia coli and decreased phosphate solubilization. These findings indicate that QS in B. plantarii significantly influences various pathogenicity and survival traits, including motility, biofilm formation, antibacterial activity, and nutrient acquisition, highlighting the critical role of QS in pathogen virulence and adaptability.

Evaluating the effects of virulence genotype, swarming motility, and multi-locus sequence types of Escherichia coli on layer chicken embryos.

To determine the effects of swarming motility (SM) and multi-locus sequence types (MLST) on the main effect of virulence genotype of Escherichia coli through an embryos lethality assay between the 12th and 18th days of incubation. We collected 58 E. coli isolates from asymptomatic commercial hens (n=42) and lesions of colibacillosis cases (n=16), then classified their virulence genotype as avirulent, moderately virulent, virulent-healthy, and virulent-colibacillosis categories by the presence of five virulence-associated genes (iroN, ompT, hlyF, iutA, and iss). These isolates were further classified as non-motile, motile, or hyper-motile by SM assay. From the 58 isolates, we selected 29 for ELA and determined their MLST. Each isolate was inoculated into 15 embryonated eggs through the allantoic cavity. We found the avirulent isolates reduced the relative embryo weight compared to virulent-colibacillosis and moderately virulent isolates (37.49vs. 41.51 and 40.34%, P=0.03). Among the moderately virulent and virulent-colibacillosis categories, embryo lethality was lower when isolates were non-motile. Yolk retention was unaffected by virulence categories, motility, or MLST. Interaction between virulence genotype and SM substantially influenced the embryo lethality assay of E. coli isolates.

Comparative genomic analysis of strain Priestia megaterium B1 reveals conserved potential for adaptation to endophytism and plant growth promotion.

In our study, we aimed to explore the genomic and phenotypic traits of Priestia megaterium strain B1, which was isolated from root material of healthy apple plants, to adapt to the endophytic lifestyle and promote plant growth. We identified putative genes encoding proteins involved in chemotaxis, flagella biosynthesis, biofilm formation, secretory systems, detoxification, transporters, and transcription regulation. Furthermore, B1 exhibited both swarming and swimming motilities, along with biofilm formation. Both genomic and physiological analyses revealed the potential of B1 to promote plant growth through the production of indole-3-acetic acid and siderophores, as well as the solubilization of phosphate and zinc. To deduce potential genomic features associated with endophytism across members of P. megaterium strains, we conducted a comparative genomic analysis involving 27 and 31 genomes of strains recovered from plant and soil habitats, respectively, in addition to our strain B1. Our results indicated a closed pan genome and comparable genome size of strains from both habitats, suggesting a facultative host association and adaptive lifestyle to both habitats. Additionally, we performed a sparse Partial Least Squares Discriminant Analysis to infer the most discriminative functional features of the two habitats based on Pfam annotation. Despite the distinctive clustering of both groups, functional enrichment analysis revealed no significant enrichment of any Pfam domain in both habitats. Furthermore, when assessing genetic elements related to adaptation to endophytism in each individual strain, we observed their widespread presence among strains from both habitats. Moreover, all members displayed potential genetic elements for promoting plant growth.IMPORTANCEBoth genomic and phenotypic analyses yielded valuable insights into the capacity of P. megaterium B1 to adapt to the plant niche and enhance its growth. The comparative genomic analysis revealed that P. megaterium members, whether derived from soil or plant sources, possess the essential genetic machinery for interacting with plants and enhancing their growth. The conservation of these traits across various strains of this species extends its potential application as a bio-stimulant in diverse environments. This significance also applies to strain B1, particularly regarding its application to enhance the growth of plants facing apple replant disease conditions.

Prunus persica leaves aqueous extract mediated biosynthesis of Ag nanoparticles and assessment of its anti-quorum sensing potential against Hafnia species.

Hafnia sp. was one of the specific spoilage bacteria in aquatic products, and the aim of the study was to investigate the inhibition ability of the silver nanoparticles (AgNPs) biosynthesis by an aqueous extract of Prunus persica leaves toward the spoilage-related virulence factors of Hafnia sp. The synthesized P-AgNPs were spherical, with a mean particle size of 36.3nm and zeta potential of 21.8±1.33mV. In addition, the inhibition effects of P-AgNPs on the growth of two Hafnia sp. strains and their quorum sensing regulated virulence factors, such as the formation of biofilm, secretion of N-acetyl-homoserine lactone (AHLs), proteases, and exopolysaccharides, as well as their swarming and swimming motilities were evaluated. P-AgNPs had a minimum inhibitory concentration (MIC) of 64 μg ml-1 against the two Hafnia sp. strains. When the concentration of P-AgNPs was below MIC, it could inhibit the formation of biofilms by Hafnia sp at 8-32 μg ml-1, but it promoted the formation of biofilms by Hafnia sp at 0.5-4 μg ml-1. P-AgNPs exhibited diverse inhibiting effects on AHLs and protease production, swimming, and swarming motilities at various concentrations.

Multiple pathways impact the swarming motility of Pseudomonas fluorescens Pf0-1.

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 Pseudomonas fluorescens Pf0-1 is swarming deficient due to a point mutation in the gacA 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 P. fluorescens Pf0-1. Here, we demonstrate that a ΔrsmA ΔrsmE ΔrsmI 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 ΔrsmA ΔrsmE ΔrsmI 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 P. fluorescens 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 P. fluorescens 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 via 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.

The Anti-Microbial Peptide Citrocin Controls Pseudomonas aeruginosa Biofilms by Breaking Down Extracellular Polysaccharide.

Citrocin is an anti-microbial peptide that holds great potential in animal feed. This study evaluates the anti-microbial and anti-biofilm properties of Citrocin and explores the mechanism of action of Citrocin on the biofilm of P. aeruginosa. The results showed that Citrocin had a significant inhibitory effect on the growth of P. aeruginosa with a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 0.3 mg/mL. All five concentrations (1/4MIC, 1/2MIC, MIC, 2MIC, and 4MIC) of Citrocin inhibited P. aeruginosa biofilm formation. Citrocin at the MIC, 2MIC and 4MIC removed 42.7%, 76.0% and 83.2% of mature biofilms, respectively, and suppressed the swarming motility, biofilm metabolic activity and extracellular polysaccharide production of P. aeruginosa. Metabolomics analysis indicated that 0.3 mg/mL of Citrocin up- regulated 26 and down-regulated 83 metabolites, mainly comprising amino acids, fatty acids, organic acids and sugars. Glucose and amino acid metabolic pathways, including starch and sucrose metabolism as well as arginine and proline metabolism, were highly enriched by Citrocin. In summary, our research reveals the anti-biofilm mechanism of Citrocin at the metabolic level, which provides theoretical support for the development of novel anti-biofilm strategies for combatting P. aeruginosa.

Environmental magnesium ion affects global gene expression, motility, biofilm formation and virulence of Vibrio parahaemolyticus

Environmental magnesium ion affects global gene expression, motility, biofilm formation and virulence of Vibrio parahaemolyticus

Evaluation of the Antioxidant, Cytotoxicity, Antibacterial, Anti-Motility, and Anti-Biofilm Effects of Myrothamnus flabellifolius Welw. Leaves and Stem Defatted Subfractions.

The formation of biofilms underscores the challenge of treating bacterial infections. The study aimed to assess the antioxidant, cytotoxicity, antibacterial, anti-motility, and anti-biofilm effects of defatted fractions from Myrothamnus flabellifolius (resurrection plant). Antioxidant activity was assessed using DPPH radical scavenging and hydrogen peroxide assays. Cytotoxicity was screened using a brine shrimp lethality assay. Antibacterial activity was determined using the micro-dilution and growth curve assays. Antibiofilm potential was screened using the crystal violet and tetrazolium reduction assay. Liquid-liquid extraction of crude extracts concentrated polyphenols in the ethyl acetate and n-butanol fractions. Subsequently, these fractions had notable antioxidant activity and demonstrated broad-spectrum antibacterial activity against selected Gram-negative and Gram-positive bacteria and Mycobacterium smegmatis (MIC values < 630 μg/mL). Growth curves showed that the bacteriostatic inhibition by the ethyl acetate fractions was through the extension of the lag phase and/or suppression of the growth rate. The sub-inhibitory concentrations of the ethyl acetate fractions inhibited the swarming motility of Pseudomonas aeruginosa and Klebsiella pneumoniae by 100% and eradicated more than 50% of P. aeruginosa biofilm biomass. The polyphenolic content of M. flabellifolius plays an important role in its antibacterial, anti-motility, and antibiofilm activity, thus offering an additional strategy to treat biofilm-associated infections.

An atypical 3-ketoacyl ACP synthase III required for acyl homoserine lactone synthesis in Pseudomonas syringae pv. syringae B728a.

The last step of the initiation phase of fatty acid biosynthesis in most bacteria is catalyzed by the 3-ketoacyl-acyl carrier protein (ACP) synthase III (FabH). Pseudomonas syringae pv. syringae strain B728a encodes two FabH homologs, Psyr_3467 and Psyr_3830, which we designated PssFabH1 and PssFabH2, respectively. Here, we explored the roles of these two 3-ketoacyl-ACP synthase (KAS) III proteins. We found that PssFabH1 is similar to the Escherichia coli FabH in using acetyl-acetyl-coenzyme A (CoA ) as a substrate in vitro, whereas PssFabH2 uses acyl-CoAs (C4-C10) or acyl-ACPs (C6-C10). Mutant analysis showed that neither KAS III protein is essential for the de novo fatty acid synthesis and cell growth. Loss of PssFabH1 reduced the production of an acyl homoserine lactone (AHL) quorum-sensing signal, and this production was partially restored by overexpressing FabH homologs from other bacteria. AHL production was also restored by inhibiting fatty acid elongation and providing exogenous butyric acid. Deletion of PssFabH1 supports the redirection of acyl-ACP toward biosurfactant synthesis, which in turn enhances swarming motility. Our study revealed that PssFabH1 is an atypical KAS III protein that represents a new KAS III clade that functions in providing a critical fatty acid precursor, butyryl-ACP, for AHL synthesis.IMPORTANCEAcyl homoserine lactones (AHLs) are important quorum-sensing compounds in Gram-negative bacteria. Although their formation requires acylated acyl carrier proteins (ACPs), how the acylated intermediate is shunted from cellular fatty acid synthesis to AHL synthesis is not known. Here, we provide in vivo evidence that Pseudomonas syringae strain B728a uses the enzyme PssFabH1 to provide the critical fatty acid precursor butyryl-ACP for AHL synthesis. Loss of PssFabH1 reduces the diversion of butyryl-ACP to AHL, enabling the accumulation of acyl-ACP for synthesis of biosurfactants that contribute to bacterial swarming motility. We report that PssFabH1 and PssFabH2 each encode a 3-ketoacyl-acyl carrier protein synthase (KAS) III in P. syringae B728a. Whereas PssFabH2 is able to function in redirecting intermediates from β-oxidation to fatty acid synthesis, PssFabH1 is an atypical KAS III protein that represents a new KAS III clade based on its sequence, non-involvement in cell growth, and novel role in AHL synthesis.

Insights into the complementation potential of the extreme acidophile's orthologue in replacing Escherichia coli hfq gene-particularly in bacterial resistance to environmental stress.

Acidithiobacillus caldus is a typical extreme acidophile widely used in the biohydrometallurgical industry, which often experiences extreme environmental stress in its natural habitat. Hfq, an RNA-binding protein, typically functions as a global regulator involved in various cellular physiological processes. Yet, the biological functions of Hfq derived from such extreme acidophile have not been extensively investigated. In this study, the recombinant strain Δhfq/Achfq, constructed by CRISPR/Cas9-mediated chromosome integration, fully or partially restored the phenotypic defects caused by hfq deletion in Escherichia coli, including impaired growth performance, abnormal cell morphology, impaired swarming motility, decreased stress resistance, decreased intracellular ATP and free amino acid levels, and attenuated biofilm formation. Particularly noteworthy, the intracellular ATP level and biofilm production of the recombinant strain were increased by 12.2% and 7.0%, respectively, compared to the Δhfq mutant. Transcriptomic analysis revealed that even under heterologous expression, AcHfq exerted global regulatory effects on multiple cellular processes, including metabolism, environmental signal processing, and motility. Finally, we established a potential working model to illustrate the regulatory mechanism of AcHfq in bacterial resistance to environmental stress.