Type 1 and type 2 strains of Mycoplasma pneumoniae form different biofilms

Simmons, Warren L.; Daubenspeck, James M.; Osborne, John D.; Balish, Mitchell F.; Waites, Ken B.; Dybvig, Kevin

doi:10.1099/mic.0.064782-0

Cited by 67 publications

(85 citation statements)

References 45 publications

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“…2a, P < 0.01). Strains of mycoplasma forming weak biofilms typically grow well but have a higher percentage of CFU that are planktonic than do strains of mycoplasmas that form more robust biofilms [11,10]. As the crystal violet assay was limited to assessing cells within the biofilm, we determined the amount of protein adhered to the sealed flasks as well as the protein from the unadhered cells in the medium.…”

Section: Resultsmentioning

confidence: 99%

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Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae

Simmons

Dybvig

2015

Curr Microbiol

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Mycoplasma pneumoniae causes chronic respiratory disease in humans. Factors thought to be important for colonization include the ability of the mycoplasma to form a biofilm on epithelial surfaces and the production of hydrogen peroxide to damage host tissue. Almost all of the mycoplasmas, including M. pneumoniae, lack superoxide dismutase and catalase and a balance should exist between peroxide production and growth. We show here that the addition of catalase to cultures enhanced the formation of biofilms and altered the structure. The incorporation of catalase in agar increased the number of colony-forming units detected and hence could improve the clinical diagnosis of mycoplasmal diseases.

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Section: Resultsmentioning

confidence: 99%

“…M. pneumoniae strain UAB_PO1 was assayed as CFU on 60 mm plates containing 10 ml SP-4 agar as described [10]. Plates were either left unsealed or were sealed with several layers of parafilm to minimize gas exchange.…”

Section: Methodsmentioning

confidence: 99%

Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae

Simmons

Dybvig

2015

Curr Microbiol

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“…In M. pulmonis, EPS-I contains equimolar amounts of glucose and galactose, with undetermined linkage (24), while EPS-II, involved in biofilm formation, and another polysaccharide of M. pneumoniae contain also N-acetylglucosamine (25). Members of the MMC differ from M. pneumoniae and M. pulmonis and appear to be diverse in terms of polysaccharide production and secretion in spite of their very close phylogenetic relationship.…”

Section: Discussionmentioning

confidence: 99%

Highly Dynamic Genomic Loci Drive the Synthesis of Two Types of Capsular or Secreted Polysaccharides within the Mycoplasma mycoides Cluster

Bertin

Pau-Roblot

Courtois

et al. 2015

Appl Environ Microbiol

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iMycoplasmas of the Mycoplasma mycoides cluster are all ruminant pathogens. Mycoplasma mycoides subsp. mycoides is responsible for contagious bovine pleuropneumonia and is known to produce capsular polysaccharide (CPS) and exopolysaccharide (EPS). Previous studies have strongly suggested a role for Mycoplasma mycoides subsp. mycoides polysaccharides in pathogenicity. Mycoplasma mycoides subsp. mycoides-secreted EPS was recently characterized as a ␤(1¡6)-galactofuranose homopolymer (galactan) identical to the capsular product. Here, we extended the characterization of secreted polysaccharides to all other members of the M. mycoides cluster: M. capricolum subsp. capripneumoniae, M. capricolum subsp. capricolum, M. leachii, and M. mycoides subsp. capri (including the LC and Capri serovars). Extracted EPS was characterized by nuclear magnetic resonance, resulting in the identification of a homopolymer of ␤(1¡2)-glucopyranose (glucan) in M. capricolum subsp. capripneumoniae and M. leachii. Monoclonal antibodies specific for this glucan and for the Mycoplasma mycoides subsp. mycoides-secreted galactan were used to detect the two polysaccharides. While M. mycoides subsp. capri strains of serovar LC produced only capsular galactan, no polysaccharide could be detected in strains of serovar Capri. All strains of M. capricolum subsp. capripneumoniae and M. leachii produced glucan CPS and EPS, whereas glucan production and localization varied among M. capricolum subsp. capricolum strains. Genes associated with polysaccharide synthesis and forming a biosynthetic pathway were predicted in all cluster members. These genes were organized in clusters within two loci representing genetic variability hot spots. Phylogenetic analysis showed that some of these genes, notably galE and glf, were acquired via horizontal gene transfer. These findings call for a reassessment of the specificity of the serological tests based on mycoplasma polysaccharides. Within the class Mollicutes, the so-called Mycoplasma mycoides cluster (MMC) (1) is unique: all the members of the MMC are pathogenic for ruminants, yet from a phylogenetic point of view, this cluster belongs to the Spiroplasma clade, which includes a number of species isolated from plants or insects (2). The taxonomy of the MMC was modified recently to reflect more precisely the phylogeny of this group. Because all the cluster members are closely related, their 16S rRNA gene sequences did not provide sufficient resolution to discriminate them accurately (3). A multilocus sequence typing approach was used to obtain a more precise phylogeny (4), which led to a simplification of the taxonomy (5) ( Table 1). The cluster currently comprises five species or subspecies: Mycoplasma mycoides subsp. mycoides, which was known formerly as the "small colony" (SC) biotype (1), M. mycoides subsp. capri, M. capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae, and M. leachii. In addition, M. mycoides subsp. capri is subdivided in two serovars: the "large colony" serovar (M. mycoides subsp...

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“…The current list of mycoplasmas producing CPS or EPS is not exhaustive, however, as purification of polysaccharides has been hampered for many years by the use of complex culture media. A detailed description of the polysaccharides secreted by nonruminant mycoplasmas is available only for Mycoplasma pulmonis and M. pneumoniae (10,11).…”

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Mycoplasma agalactiae Secretion of β-(1→6)-Glucan, a Rare Polysaccharide in Prokaryotes, Is Governed by High-Frequency Phase Variation

Gaurivaud

Baranowski

Pau-Roblot

et al. 2016

Appl Environ Microbiol

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Mycoplasmas are minimal, wall-less bacteria but have retained the ability to secrete complex carbohydrate polymers that constitute a glycocalyx. In members of the Mycoplasma mycoides cluster, which are important ruminant pathogens, the glycocalyx includes both cell-attached and cell-free polysaccharides. This report explores the potential secretion of polysaccharides by M. agalactiae, another ruminant pathogen that belongs to a distant phylogenetic group. Comparative genomic analyses showed that M. agalactiae possesses all the genes required for polysaccharide secretion. Notably, a putative synthase gene (gsmA) was identified, by in silico reconstruction of the biosynthetic pathway, that could be involved in both polymerization and export of the carbohydrate polymers. M. agalactiae polysaccharides were then purified in vitro and found to be mainly cell attached, with a linear ␤-(1¡6)-glucopyranose structure [␤-(1¡6)-glucan]. Secretion of ␤-(1¡6)-glucan was further shown to rely on the presence of a functional gsmA gene, whose expression is subjected to high-frequency phase variation. This event is governed by the spontaneous intraclonal variation in length of a poly(G) tract located in the gsmA coding sequence and was shown to occur in most of the M. agalactiae clinical isolates tested in this study. M. agalactiae susceptibility to serum-killing activity appeared to be dictated by ON/OFF switching of ␤-(1¡6)-glucan secretion, suggesting a role of this phenomenon in survival of the pathogen when it invades the host bloodstream. Finally, ␤-(1¡6)-glucan secretion was not restricted to M. agalactiae but was detected also in M. mycoides subsp. capri PG3 T , another pathogen of small ruminants. IMPORTANCEMany if not all bacteria are able to secrete polysaccharides, either attached to the cell surface or exported unbound into the extracellular environment. Both types of polysaccharides can play a role in bacterium-host interactions. Mycoplasmas are no exception despite their poor overall metabolic capacity. We showed here that M. agalactiae secretes a capsular ␤-(1¡6)-glucopyranose thanks to a specific glycosyltransferase with synthase activity. This secretion is governed by high-frequency ON/OFF phase variation that might be crucial in mycoplasma host dissemination, as cell-attached ␤-(1¡6)-glucopyranose increases serum-killing susceptibility. Our results provide functional genetic data about mycoplasmal glycosyltransferases with dual functions, i.e., assembly and export of the sugar polymers across the cell membrane. Furthermore, we demonstrated that nonprotein epitopes can be subjected to surface antigenic variation in mycoplasmas. Finally, the present report contributes to unravel the role of secreted polysaccharides in the virulence and pathogenicity of these peculiar bacteria.

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Type 1 and type 2 strains of Mycoplasma pneumoniae form different biofilms

Cited by 67 publications

References 45 publications

Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae

Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae

Highly Dynamic Genomic Loci Drive the Synthesis of Two Types of Capsular or Secreted Polysaccharides within the Mycoplasma mycoides Cluster

Mycoplasma agalactiae Secretion of β-(1→6)-Glucan, a Rare Polysaccharide in Prokaryotes, Is Governed by High-Frequency Phase Variation

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