scholarly journals Loss of a Biofilm-Inhibiting Glycosyl Hydrolase during the Emergence of Yersinia pestis

2008 ◽  
Vol 190 (24) ◽  
pp. 8163-8170 ◽  
Author(s):  
David L. Erickson ◽  
Clayton O. Jarrett ◽  
Julie A. Callison ◽  
Elizabeth R. Fischer ◽  
B. Joseph Hinnebusch
Keyword(s):  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
Glycosyl Hydrolase ◽  
Extracellular Matrices ◽  
Bacterial Agent ◽  
Glycosidase Activity ◽  
By Products ◽  
Transmissible Infection

ABSTRACT Yersinia pestis, the bacterial agent of plague, forms a biofilm in the foregut of its flea vector to produce a transmissible infection. The closely related Yersinia pseudotuberculosis, from which Y. pestis recently evolved, can colonize the flea midgut but does not form a biofilm in the foregut. Y. pestis biofilm in the flea and in vitro is dependent on an extracellular matrix synthesized by products of the hms genes; identical genes are present in Y. pseudotuberculosis. The Yersinia Hms proteins contain functional domains present in Escherichia coli and Staphylococcus proteins known to synthesize a poly-β-1,6-N-acetyl-d-glucosamine biofilm matrix. In this study, we show that the extracellular matrices (ECM) of Y. pestis and staphylococcal biofilms are antigenically related, indicating a similar biochemical structure. We also characterized a glycosyl hydrolase (NghA) of Y. pseudotuberculosis that cleaved β-linked N-acetylglucosamine residues and reduced biofilm formation by staphylococci and Y. pestis in vitro. The Y. pestis nghA ortholog is a pseudogene, and overexpression of functional nghA reduced ECM surface accumulation and inhibited the ability of Y. pestis to produce biofilm in the flea foregut. Mutational loss of this glycosidase activity in Y. pestis may have contributed to the recent evolution of flea-borne transmission.

scholarly journals Serotype Differences and Lack of Biofilm Formation Characterize Yersinia pseudotuberculosis Infection of the Xenopsylla cheopis Flea Vector of Yersinia pestis

2006 ◽  
Vol 188 (3) ◽  
pp. 1113-1119 ◽  
Author(s):  
David L. Erickson ◽  
Clayton O. Jarrett ◽  
Brendan W. Wren ◽  
B. Joseph Hinnebusch
Keyword(s):  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
Ex Vivo ◽  
Blood Feeding ◽  
Genetic Change ◽  
Xenopsylla Cheopis ◽  
C Elegans ◽  
Transmissible Infection

ABSTRACT Yersinia pestis, the agent of plague, is usually transmitted by fleas. To produce a transmissible infection, Y. pestis colonizes the flea midgut and forms a biofilm in the proventricular valve, which blocks normal blood feeding. The enteropathogen Yersinia pseudotuberculosis, from which Y. pestis recently evolved, is not transmitted by fleas. However, both Y. pestis and Y. pseudotuberculosis form biofilms that adhere to the external mouthparts and block feeding of Caenorhabditis elegans nematodes, which has been proposed as a model of Y. pestis-flea interactions. We compared the ability of Y. pestis and Y. pseudotuberculosis to infect the rat flea Xenopsylla cheopis and to produce biofilms in the flea and in vitro. Five of 18 Y. pseudotuberculosis strains, encompassing seven serotypes, including all three serotype O3 strains tested, were unable to stably colonize the flea midgut. The other strains persisted in the flea midgut for 4 weeks but did not increase in numbers, and none of the 18 strains colonized the proventriculus or produced a biofilm in the flea. Y. pseudotuberculosis strains also varied greatly in their ability to produce biofilms in vitro, but there was no correlation between biofilm phenotype in vitro or on the surface of C. elegans and the ability to colonize or block fleas. Our results support a model in which a genetic change in the Y. pseudotuberculosis progenitor of Y. pestis extended its pre-existing ex vivo biofilm-forming ability to the flea gut environment, thus enabling proventricular blockage and efficient flea-borne transmission.

scholarly journals Identification of gmhA, a Yersinia pestis Gene Required for Flea Blockage, by Using a Caenorhabditis elegans Biofilm System

2005 ◽  
Vol 73 (11) ◽  
pp. 7236-7242 ◽  
Author(s):  
Creg Darby ◽  
Sandya L. Ananth ◽  
Li Tan ◽  
B. Joseph Hinnebusch
Keyword(s):  
Caenorhabditis Elegans ◽  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
C Elegans ◽  
Transposon Insertion ◽  
Insertion Mutants ◽  
Random Screening

ABSTRACT Yersinia pestis, the cause of bubonic plague, blocks feeding by its vector, the flea. Recent evidence indicates that blockage is mediated by an in vivo biofilm. Y. pestis and the closely related Yersinia pseudotuberculosis also make biofilms on the cuticle of the nematode Caenorhabditis elegans, which block this laboratory animal's feeding. Random screening of Y. pseudotuberculosis transposon insertion mutants with a C. elegans biofilm assay identified gmhA as a gene required for normal biofilms. gmhA encodes phosphoheptose isomerase, an enzyme required for synthesis of heptose, a conserved component of lipopolysaccharide and lipooligosaccharide. A Y. pestis gmhA mutant was constructed and was severely defective for C. elegans biofilm formation and for flea blockage but only moderately defective in an in vitro biofilm assay. These results validate use of the C. elegans biofilm system to identify genes and pathways involved in Y. pestis flea blockage.

scholarly journals High-Frequency RecA-Dependent and -Independent Mechanisms of Congo Red Binding Mutations in Yersinia pestis

1999 ◽  
Vol 181 (16) ◽  
pp. 4896-4904 ◽  
Author(s):  
Janelle M. Hare ◽  
Kathleen A. McDonough
Keyword(s):  
Yersinia Pestis ◽  
Congo Red ◽  
High Frequency ◽  
Yersinia Pseudotuberculosis ◽  
Resistance Mutation ◽  
Laboratory Culture ◽  
Uptake System ◽  
Deletion Event ◽  
Genes Encoding

ABSTRACT Yersinia pestis, which causes bubonic and pneumonic plague, forms pigmented red colonies on Congo red (CR) dye agar. ThehmsHFRS genes required for CR binding (Crb+) are genetically linked to virulence-associated genes encoding a siderophore uptake system. These genes are contained in a 102-kb chromosomal pgm locus that is lost in a high-frequency deletion event, resulting in loss of the Crb+ phenotype. We constructed a recA mutant strain of Y. pestisKIM10+ (YPRA) to test whether the high frequency Crb mutants result from a RecA-mediated deletion of the IS100-flankedpgm locus. Two Pgm-associated phenotypes (Crb+and pesticin sensitivity [Psts]) were used as markers for the presence of the pgm locus in the RecA+KIM10+ and RecA− YPRA strains. In KIM10+, both phenotypes were lost at a very high (2 × 10−3) frequency, due to the deletion of the entire pgm locus. In YPRA, the Crb+ phenotype was still lost at a high frequency (4.5 × 10−5), although the loss of the Pstsphenotype occurred at spontaneous antibiotic resistance mutation frequencies (2 × 10−7). These RecA-independent Crb− mutants were caused by mutations in both thehmsHFRS locus and in a newly identified gene,hmsT. Nonpigmented Yersinia pseudotuberculosisand Escherichia coli strains transformed with bothhmsT and hmsHFRS became Crb+. This study demonstrates that in a laboratory culture, the Crb+phenotype is unstable, independent of the pgm locus deletion. We propose that a lack of selection for the CR-binding ability of Y. pestis in vitro may contribute to the mutation frequencies observed at the hmsHFRS andhmsT loci.

scholarly journals Differential Gene Expression Patterns ofYersinia pestisandYersinia pseudotuberculosisduring Infection and Biofilm Formation in the Flea Digestive Tract

mSystems ◽  
2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Iman Chouikha ◽  
Daniel E. Sturdevant ◽  
Clayton Jarrett ◽  
Yi-Cheng Sun ◽  
B. Joseph Hinnebusch
Keyword(s):  
Digestive Tract ◽  
Biofilm Formation ◽  
Yersinia Pseudotuberculosis ◽  
Expression Patterns ◽  
Type Vi Secretion System ◽  
Genetic Changes ◽  
Content Type ◽  
Biofilm Development

ABSTRACTYersinia pestis, the etiologic agent of plague, emerged as a fleaborne pathogen only within the last 6,000 years. Just five simple genetic changes in theYersinia pseudotuberculosisprogenitor, which served to eliminate toxicity to fleas and to enhance survival and biofilm formation in the flea digestive tract, were key to the transition to the arthropodborne transmission route. To gain a deeper understanding of the genetic basis for the development of a transmissible biofilm infection in the flea foregut, we evaluated additional gene differences and performedin vivotranscriptional profiling ofY. pestis, aY. pseudotuberculosiswild-type strain (unable to form biofilm in the flea foregut), and aY. pseudotuberculosismutant strain (able to produce foregut-blocking biofilm in fleas) recovered from fleas 1 day and 14 days after an infectious blood meal. Surprisingly, theY. pseudotuberculosismutations that increased c-di-GMP levels and enabled biofilm development in the flea did not change the expression levels of thehmsgenes responsible for the synthesis and export of the extracellular polysaccharide matrix required for mature biofilm formation. TheY. pseudotuberculosismutant uniquely expressed much higher levels ofYersiniatype VI secretion system 4 (T6SS-4) in the flea, and this locus was required for flea blockage byY. pseudotuberculosisbut not for blockage byY. pestis. Significant differences between the two species in expression of several metabolism genes, the Psa fimbrial genes, quorum sensing-related genes, transcription regulation genes, and stress response genes were evident during flea infection.IMPORTANCEY. pestisemerged as a highly virulent, arthropod-transmitted pathogen on the basis of relatively few and discrete genetic changes fromY. pseudotuberculosis. Parallel comparisons of thein vitroandin vivotranscriptomes ofY. pestisand twoY. pseudotuberculosisvariants that produce a nontransmissible infection and a transmissible infection of the flea vector, respectively, provided insights into howY. pestishas adapted to life in its flea vector and point to evolutionary changes in the regulation of metabolic and biofilm development pathways in these two closely related species.

Abstract 90: Serum Kawasaki Disease-specific Microbe-associated Molecular Patterns are Derived from in Vitro and in Vivo Biofilms

Circulation ◽  
2015 ◽  
Vol 131 (suppl_2) ◽  
Author(s):  
Takeshi Kusuda ◽  
Yasutaka Nakashima ◽  
Murata Kenji ◽  
Shunsuke Kanno ◽  
Mitsumasa Saito ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Yersinia Pseudotuberculosis ◽  
Molecular Structures ◽  
Human Coronary Artery ◽  
Medium Temperature ◽  
Molecular Patterns ◽  
Culture Supernatants

Background: We have found that serum KD-specific molecules possess molecular structures common to microbe-associated molecular patterns (MAMPs) from several bacteria by liquid chromatography-mass spectrometry (LC-MS) analysis. In the present study, we have extensively searched for the culture conditions that reproducibly produce MAMPs common to serum KD specific molecules. Methods: Yersinia pseudotuberculosis, spore-forming bacteria and pathogenic bacteria were obtained from KD patients. Others were purchased from American Type Culture Collection. Microbes were cultured in a variety of conditions (medium, temperature, duration, shaking and biofilm formation). For biofilm formation, a various kind of oil/butter was added to the culture medium. Extraction from in vitro samples (culture supernatants, bacteria cells, and biofilms) as well as in vivo biofilm samples (teeth, tongue, nasal cavity, or rectum) was performed with ethyl acetate. Samples were analyzed by HPLC and MS (Esquire 6000 electrospray ionization). Human coronary artery endothelial cells (HCAECs) were employed for cytokine assay. The concentrations of cytokines were measured by EC800 cell analyzer with a BD™ Cytometric Bead Array. Results: Serum KD-specific molecules showed m/z and MS/MS fragmentation patterns almost identical to those of MAMPs obtained from the biofilms formed in vitro (Bacillus cereus, Bacillus subtilis, Yersinia pseudotuberculosis, Staphylococcus aureus) or in vivo. Not culture supernatants but biofilm extracts from these bacteria, especially cultured in the presence of butter, induced proinflammatory cytokines by HCAECs. Conclusions: Extensive analysis by LC-MS/MS revealed that serum KD-specific molecules common to MAMPs were mostly derived from biofilms in vitro and in vivo. This report offers novel insight into the pathogenesis of KD.

scholarly journals Differential Control of Yersinia pestis Biofilm Formation In Vitro and in the Flea Vector by Two c-di-GMP Diguanylate Cyclases

PLoS ONE ◽  
2011 ◽  
Vol 6 (4) ◽  
pp. e19267 ◽  
Author(s):  
Yi-Cheng Sun ◽  
Alexandra Koumoutsi ◽  
Clayton Jarrett ◽  
Kevin Lawrence ◽  
Frank C. Gherardini ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Biofilm Formation ◽  
Diguanylate Cyclases ◽  
Differential Control

scholarly journals Polyphenols Content and In Vitro α-Glycosidase Activity of Different Italian Monofloral Honeys, and Their Effect on Selected Pathogenic and Probiotic Bacteria

2021 ◽  
Vol 9 (8) ◽  
pp. 1694
Author(s):  
Florinda Fratianni ◽  
Maria Neve Ombra ◽  
Antonio d’Acierno ◽  
Lucia Caputo ◽  
Giuseppe Amato ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Probiotic Bacteria ◽  
Polyphenol Content ◽  
Total Polyphenols ◽  
E Coli ◽  
Glycosidase Activity ◽  
Probiotic Strains ◽  
Italian Origin ◽  
And Staphylococcus Aureus

We evaluated the polyphenol content and the α-glucosidase activity exhibited by different monofloral honeys of Italian origin. Their capacity to act on different pathogenic (Acinetobacter baumannii, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus) as well as probiotic bacteria (Lacticaseibacillus casei, Lactobacillus acidophilus, Lactiplantibacillus plantarum, Lactobacillus gasseri, and Lacticaseibacillus rhamnosus) was also assessed. Total polyphenols varied between 110.46 μg/g of fresh product (rhododendron honey) and 552.29 μg/g of fresh product (strawberry tree honey). Such result did not correspond to a parallel inhibitory α-glycosidase activity that, in each case was never higher than 33 μg/mL. Honeys were differently capable to fight the biofilm formation of the pathogens (inhibition up to 93.27%); they inhibited the in vitro adhesive process (inhibition up to 84.27%), and acted on mature biofilm (with values up to 76.64%). Their effect on bacterial metabolism was different too. Honeys were ineffective to inhibit E. coli mature biofilm nor to act on its metabolism. The action of the honey on probiotic strains seemed almost always stimulate their growth. Thus, these monofloral honeys might exhibit effects on human health and act positively as prebiotics.

scholarly journals Poly-N-Acetylglucosamine Is Not a Major Component of the Extracellular Matrix in Biofilms Formed by icaADBC-Positive Staphylococcus lugdunensis Isolates

2007 ◽  
Vol 75 (10) ◽  
pp. 4728-4742 ◽  
Author(s):  
Kristi L. Frank ◽  
Robin Patel
Keyword(s):  
Extracellular Matrix ◽  
Wheat Germ Agglutinin ◽  
Biofilm Formation ◽  
Wheat Germ ◽  
Glycosyl Hydrolase ◽  
Joint Infection ◽  
Common Mechanism ◽  
Staphylococcus Lugdunensis ◽  
Sodium Metaperiodate

ABSTRACT Staphylococcus lugdunensis is a pathogen of heightened virulence that causes infections resembling those caused by Staphylococcus aureus rather than those caused by its coagulase-negative staphylococcal counterparts. Many types of S. lugdunensis infection, including native valve endocarditis, prosthetic joint infection, and intravascular catheter-related infection, are associated with biofilm etiology. Poly-N-acetylglucosamine (PNAG), a polysaccharide synthesized by products of the icaADBC locus, is a common mechanism of intercellular adhesion in staphylococcal biofilms. Here we report the characterization of ica homologues and the in vitro biofilm formation properties of a collection of S. lugdunensis clinical isolates. Isolates formed biofilms in microtiter wells to various degrees. Biofilm formation by most isolates was enhanced with glucose but diminished by sodium chloride or ethanol. icaADBC homologues were found in all S. lugdunensis isolates tested, although the locus organization differed substantially from that of other staphylococcal ica loci. icaR was not detected in S. lugdunensis, but a novel open reading frame with putative glycosyl hydrolase function is located upstream of the ica locus. icaADBC sequence heterogeneity did not explain the variability in biofilm formation among isolates. PNAG was not detected in S. lugdunensis extracts by immunoblotting with an anti-deacetylated PNAG antibody or wheat germ agglutinin. Confocal microscopy with fluorescently labeled wheat germ agglutinin showed a paucity of PNAG in S. lugdunensis biofilms, but abundant extracellular protein was visualized with SYPRO Ruby staining. Biofilms were resistant to detachment by dispersin B and sodium metaperiodate but were susceptible to detachment by proteases. Despite the genetic presence of icaADBC homologues in S. lugdunensis isolates, PNAG is not a major component of the extracellular matrix of in vitro biofilms formed by this species. Our data suggest that the S. lugdunensis biofilm matrix contains proteinaceous factors.

scholarly journals Rhamnose Binding Protein as an Anti-Bacterial Agent—Targeting Biofilm of Pseudomonas aeruginosa

Marine Drugs ◽  
2019 ◽  
Vol 17 (6) ◽  
pp. 355 ◽  
Author(s):  
Tse-Kai Fu ◽  
Sim-Kun Ng ◽  
Yi-En Chen ◽  
Yuan-Chuan Lee ◽  
Fruzsina Demeter ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Lung Epithelial Cells ◽  
Bacterial Agent ◽  
Lung Epithelial ◽  
Biofilm Development

More than 80% of infectious bacteria form biofilm, which is a bacterial cell community surrounded by secreted polysaccharides, proteins and glycolipids. Such bacterial superstructure increases resistance to antimicrobials and host defenses. Thus, to control these biofilm-forming pathogenic bacteria requires antimicrobial agents with novel mechanisms or properties. Pseudomonas aeruginosa, a Gram-negative opportunistic nosocomial pathogen, is a model strain to study biofilm development and correlation between biofilm formation and infection. In this study, a recombinant hemolymph plasma lectin (rHPLOE) cloned from Taiwanese Tachypleus tridentatus was expressed in an Escherichia coli system. This rHPLOE was shown to have the following properties: (1) Binding to P. aeruginosa PA14 biofilm through a unique molecular interaction with rhamnose-containing moieties on bacteria, leading to reduction of extracellular di-rhamnolipid (a biofilm regulator); (2) decreasing downstream quorum sensing factors, and inhibiting biofilm formation; (3) dispersing the mature biofilm of P. aeruginosa PA14 to improve the efficacies of antibiotics; (4) reducing P. aeruginosa PA14 cytotoxicity to human lung epithelial cells in vitro and (5) inhibiting P. aeruginosa PA14 infection of zebrafish embryos in vivo. Taken together, rHPLOE serves as an anti-biofilm agent with a novel mechanism of recognizing rhamnose moieties in lipopolysaccharides, di-rhamnolipid and structural polysaccharides (Psl) in biofilms. Thus rHPLOE links glycan-recognition to novel anti-biofilm strategies against pathogenic bacteria.

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