Identification of capsular antigens in Serratia marcescens

Aucken, Hazel M.; Wilkinson, Stephen G.; Pitt, T. L.

doi:10.1128/jcm.35.1.59-63.1997

Cited by 17 publications

(14 citation statements)

References 27 publications

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“…We analyzed the morphologies of wild-type, YODUd, and SPRAd spores using phase-contrast microscopy and a negative staining procedure. When the spores were negatively-stained with Indian ink [31] , which is a stain commonly used to reveal polysaccharide capsules, a clear halo was visible around the wild-type spores, but not around the YODUd and SPRAd spores ( Figure 6A , top panels). The appearance of a halo is consistent with the presence of polysaccharides around the wild-type spore.…”

Section: Resultsmentioning

confidence: 99%

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Developmentally-Regulated Excision of the SPβ Prophage Reconstitutes a Gene Required for Spore Envelope Maturation in Bacillus subtilis

et al. 2014

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Temperate phages infect bacteria by injecting their DNA into bacterial cells, where it becomes incorporated into the host genome as a prophage. In the genome of Bacillus subtilis 168, an active prophage, SPβ, is inserted into a polysaccharide synthesis gene, spsM. Here, we show that a rearrangement occurs during sporulation to reconstitute a functional composite spsM gene by precise excision of SPβ from the chromosome. SPβ excision requires a putative site-specific recombinase, SprA, and an accessory protein, SprB. A minimized SPβ, where all the SPβ genes were deleted, except sprA and sprB, retained the SPβ excision activity during sporulation, demonstrating that sprA and sprB are necessary and sufficient for the excision. While expression of sprA was observed during vegetative growth, sprB was induced during sporulation and upon mitomycin C treatment, which triggers the phage lytic cycle. We also demonstrated that overexpression of sprB (but not of sprA) resulted in SPβ prophage excision without triggering the lytic cycle. These results suggest that sprB is the factor that controls the timing of phage excision. Furthermore, we provide evidence that spsM is essential for the addition of polysaccharides to the spore envelope. The presence of polysaccharides on the spore surface renders the spore hydrophilic in water. This property may be beneficial in allowing spores to disperse in natural environments via water flow. A similar rearrangement occurs in Bacillus amyloliquefaciens FZB42, where a SPβ-like element is excised during sporulation to reconstitute a polysaccharide synthesis gene, suggesting that this type of gene rearrangement is common in spore-forming bacteria because it can be spread by phage infection.

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Section: Resultsmentioning

confidence: 99%

“…B. subtilis spores were negatively stained with Indian ink, as previously described [31] with a slight modification. The purified spores were resuspended in DDW, and 2 µl of the suspension was mixed with an equivalent volume of Indian ink (Daiso Sogyo, Japan) on a slide glass.…”

Section: Methodsmentioning

confidence: 99%

Developmentally-Regulated Excision of the SPβ Prophage Reconstitutes a Gene Required for Spore Envelope Maturation in Bacillus subtilis

et al. 2014

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“…Phage-resistant acapsular variants of 11 of the 29 reference strains had been produced for a previous study (Aucken et al , 1997). These variants were used here for adsorption of O antibodies from sera to the encapsulated reference strains to produce K-specific typing reagents.…”

Section: Methodsmentioning

confidence: 99%

“…Chemical analysis of polymers present in ‘ LPS ’ extracts from the reference strains has shown that the polysaccharide repeating units can be divided into neutral and acidic structures and that many of the reference strains possess both (Wilkinson, 1990). It has been suggested that the LPS 0 antigens correlate with the neutral polysaccharides (Gaston et al, 1988 ; Gaston & Pitt, 1989a) while the acidic polysaccharides have now been shown to correspond to capsular antigens (Aucken et al, 1997). The current study was undertaken to rationalize the existing 0 typing scheme and to propose a complementary capsular K typing scheme.…”

Section: Introductionmentioning

confidence: 99%

Re-evaluation of the serotypes of Serratia marcescens and separation into two schemes based on lipopolysaccharide (O) and capsular polysaccharide (K) antigens

Aucken¹,

Wilkinson

Pitt³

1998

Microbiology

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Chemical and serological analysis has revealed that many of the 29 0 serotype reference strains of Serratia marcescens contain both neutral and acidic polysaccharides which correspond to LPS 0 antigens and capsular K antigens, respectively. New 0 and K antigen typing schemes have therefore been devised, based on the known chemical structures of the surface polysaccharides of the organism. These schemes were designed to allow the specific detection of these antigens on unknown strains using ELISAs. 0 antigens were detected using whole cells cultured in broth then autoclaved to remove capsular material, while K antigens were detected using formolized whole cells which had been cultured on glycerol agar to enhance capsule production. After testing with the 29 reference strains as well as 423 distinct clinical strains, it was apparent that different aspects of chemical structure were associated with different degrees of serological reactivity and the typing schemes were modified further to accommodate this. In general, the 0 antigen repeating unit structures were chemically simple with di-or trisaccharide backbones. Serological specificity was often provided solely by the presence or absence of an 0-acetyl substituent, or a change in the linkage between two sugar residues. Five of the 0 serotypes in the new scheme were represented by 12 of the 29 reference strains, while three reference strains lacked 0 antigens altogether, resulting in the elimination of 10 of the original 0 types. In contrast, the K antigen repeating unit structures were more complex and chemically diverse, having at least four sugar residues. Three K types were each seen in two reference strains while 12 of the 29 reference strains were acapsular. Thus, the resulting schemes contain 19 0 types and 14 K types and allow the definitive serotype identification of 5. mamescens.

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“…Biochemical tests including those of growth temperature, lactic acid isomer, and carbohydrate fermentation were performed. The Indian ink method was used to visualize bacterial capsules (Aucken et al, 1996). Plasmid DNA analysis of the strains was performed according to the method of Anderson and McKay (1983).…”

Section: Characteristics Of the Mutant Strainmentioning

confidence: 99%

Binding of Mutagens to Exopolysaccharide Produced by Lactobacillus plantarum Mutant Strain 301102S

Tsuda

Hara

Miyamoto

2008

Journal of Dairy Science

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Exopolysaccharide (EPS) was produced by Lactobacillus plantarum 301102 on exposure to the mutagenic action of acridine orange and novobiocin. The biological characteristics of this mutant strain 301102S were the same as those of the parent strain, but fermented milk prepared with the mutant strain showed antimutagenic activity on 3-amino-1,4-dimethyl-5H-pyrido indole. Only EPS-bound cells of strain 301102S showed binding ability to mutagens such as heterocyclic amines, and the mutagens were inactivated by binding to EPS. The binding ability was affected by pH; the greatest percentage binding was noted at pH 8.0. Addition of Mg(2+) and sodium dodecyl sulfate, but not oxgall, inhibited the binding ability. Therefore, the binding mechanism of the EPS may consist of ion-exchange and hydrophobic bonds, and the EPS would bind mutagens in the intestine.

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Identification of capsular antigens in Serratia marcescens

Cited by 17 publications

References 27 publications

Developmentally-Regulated Excision of the SPβ Prophage Reconstitutes a Gene Required for Spore Envelope Maturation in Bacillus subtilis

Developmentally-Regulated Excision of the SPβ Prophage Reconstitutes a Gene Required for Spore Envelope Maturation in Bacillus subtilis

Re-evaluation of the serotypes of Serratia marcescens and separation into two schemes based on lipopolysaccharide (O) and capsular polysaccharide (K) antigens

Binding of Mutagens to Exopolysaccharide Produced by Lactobacillus plantarum Mutant Strain 301102S

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