Simple assay of calcium dependency for virulent plasmid-bearing clones of Yersinia enterocolitica

Bhaduri, Saumya; Turner-Jones, C.; Taylor, Maryann M.; Lachica, R. V. F.

doi:10.1128/jcm.28.4.798-800.1990

Cited by 36 publications

(43 citation statements)

References 8 publications

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“…SOMMERS corresponding Pcells (2.4 mm in diameter) ( Fig. 2 [ii] B) when grown on BHA at 37C (Table 1) as reported previously (Bhaduri et al 1990), whereas pYV/pCD-bearing colonies (KIM 5) and pYV/pCD-less colonies (Kuma) of Y. pestis were approximately the same size (1.3-1.4 mm; SD Ϯ 0.11) ( Table 1). Colony morphologies of the CV-positive and CV-negative strains of Y. enterocolitica, Y. pseudotuberculosis and Y. pestis were examined further by using a low-magnification stereomicroscope.…”

supporting

confidence: 63%

“…Brain heart infusion (BHI) broth, BHA and 0.1% peptone water were prepared as recommended by the supplier (Becton-Dickinson, Sparks, MD). Low-calcium (238 mM Ca 2+ ) BHI agarose (BHO) was prepared by adding agarose (GibcoBRL, Grand Island, NY) to final concentration of 1.2% to BHI broth supplemented with 0.1% magnesium chloride (Sigma Chemical Co., St. Louis, MO), and CR (Sigma Chemical Co.) was added to BHO at a concentration of 75 mg/mL to prepare CR-BHO as described previously (Bhaduri et al 1990;Bhaduri et al 1991). Calcium-deficient magnesium oxalate agar (MOX) was prepared by adding 20% D-galactose (Sigma Chemical Co.), 0.25 M sodium oxalate (Sigma Chemical Co.) and 0.25 M magnesium chloride (Sigma Chemical Co.) to tryptic soy agar (Becton-Dickinson) (Bhaduri et al 1990) and CR-MOX was prepared by adding 1% CR to MOX as described previously (Bhaduri et al 1991).…”

Section: Preparation Of Mediamentioning

confidence: 99%

“…Strain Kuma, a derivative of a clinical strain of Y. pestis containing a chromosomally encoded pigmentation (CR binding) virulence determinant (Pgm + ) but lacking pYV/pCD, was also used to differentiate CR-uptake encoded by the Pgm locus and pYV/pCD, respectively (Perry and Fetherston 1997) and also used as pYV/pCD-less strain. Both Y. pestis KIM 5 (Pgm -) and Y. pestis Kuma (Pgm + ) have been extensively used to study the molecular pathogenesis of this bacterium (Perry and Fetherston 1997;Brubaker 2000;Carniel 2000 (2003) pYV/pCD-positive cultures growing at 37C on BHO as described by Bhaduri et al (1990). These two strains and Kuma strain were used as negative controls for the expression of pYV/pCD phenotypes.…”

Section: Yersinia Strainsmentioning

confidence: 99%

“…The cells were incubated at 37C for 24-48 h to assess Lcr by the appearance of pinpoint 458 S. BHADURI and C.H. SOMMERS colony (size 0.36 mm) and CR-uptake (red pinpoint colony size of 0.36 mm) (Bhaduri et al 1990;Bhaduri et al 1991).…”

Section: Determination Of Pyv/pcd-associated Phenotypic Characteristicsmentioning

confidence: 99%

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DETECTION OF YERSINIA PESTIS BY COMPARISON OF VIRULENCE PLASMID (PYV/PCD)‐ASSOCIATED PHENOTYPES IN YERSINIA SPECIES*

Bhaduri¹,

Sommers

2008

Journal of Food Safety

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Consumption of meat contaminated with Yersinia pestis can cause oro‐pharyngeal plague in humans. Existing microbiological media designed for selective detection of Y. pestis in food are not satisfactory for that purpose. Expression of genetic determinants in Yersinia species including low calcium response (Lcr), colony size, crystal violet (CV) binding, Congo red (CR) uptake, autoagglutination (AA) and hydrophobicity (HP) were compared. Lcr and CV binding were detectable within 24 h at 37C in Yersinia enterocolitica and Yersinia pseudotuberculosis but at 48 h in Y. pestis. Colony size, AA, and HP characteristics were expressed in Y. pseudotuberculosis and Y. enterocolitica, but not in Y. pestis. CR uptake in Y. pestis was demonstrated only on calcium‐deficient CR‐magnesium oxalate (CR‐MOX) tryptic soy agar but was expressed on both CR‐MOX and low‐calcium agarose medium for Y. pseudotuberculosis and Y. enterocolitica. CR‐uptake phenotype by Y. pestis can be used for the detection of this pathogen in foods. PRACTICAL APPLICATIONS Yersinia pestis, Yersinia enterocolitica and Yersinia pseudotuberculosis can cause diverse human diseases. Y. enterocolitica and Y. pseudotuberculosis can cause intestinal distress when consumed in contaminated food. Y. pestis, the causative agent of bubonic plague in humans, is also found in food and can cause febrile illness in humans. Thus, food can have a significant role in the dissemination of human plague. There is a concern for risk assessors about the presence of Y. pestis in food. A virulence plasmid of 70 kb is directly involved with virulence of these pathogens. Until now, separating Y. enterocolitica and Y. pseudotuberculosis from Y. pestis required sophisticated molecular biology and immunological techniques. This problem was eliminated by analyses of virulence plasmid encoded phenotypes as simple and reliable targets for the rapid detection of Y. pestis. These classical microbiological assays should greatly assist both regulatory agencies and the food industry in the detection of Y. pestis. Further, these techniques should also greatly assist medical and public health laboratories to identify this pathogen.

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supporting

confidence: 63%

Section: Preparation Of Mediamentioning

confidence: 99%

Section: Yersinia Strainsmentioning

confidence: 99%

Section: Determination Of Pyv/pcd-associated Phenotypic Characteristicsmentioning

confidence: 99%

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DETECTION OF YERSINIA PESTIS BY COMPARISON OF VIRULENCE PLASMID (PYV/PCD)‐ASSOCIATED PHENOTYPES IN YERSINIA SPECIES*

Bhaduri¹,

Sommers

2008

Journal of Food Safety

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“…A number of studies (McDermott et al 1993;Goverde et al 1994) have shown that the choice of incubation temperature affects the growth kinetics of plasmid-bearing and plasmid-cured strains of Escherichia coli and Y. enterocolitica. In general, virulence plasmid expression in Yersinia occurs at temperatures > 30 C, this expression is usually observed in phenotypic characteristics such as calcium dependency, autoagglutination and production of outer membrane proteins (Laird and Cavanaugh 1980;Cornelis et al 1987;Bhaduri et al 1990). Goverde et al (1994) reported that virulence plasmid expression at temperatures greater than 30 C retards the growth rate of plasmid-bearing Y. enterocolitica strains as plasmid replication places an additional metabolic burden on growing cells.…”

Section: Introductionmentioning

confidence: 99%

The effect of temperature and selective agents on the growth of Yersinia enterocolitica serotype O:3 in pure culture

et al. 2000

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This study examined the individual and combined effects of the selective agents normally present in Yersinia-selective agar (i.e. cefsulodin, irgasan and novobiocin) on the growth kinetics of plasmid-bearing (P) and plasmid-cured (P±) Yersinia enterocolitica serotype O:3 at 25 and 37 C. Growth studies were carried out in pure culture, and the data obtained were subjected to linear regression analysis to determine lag phase duration(s) and growth rates of the examined strains. In general, the presence of selective agents increased the duration of the lag phase at 37 C, with longer lag phases noted in all cases in which two or more selective agents were present. Growth rates in CIN broth base (CIN NA) and CIN NA plus commercial supplement (SR 109) (CIN) were faster at 37 than 25 C, but in some cultures of incomplete CIN NA broth with less than three supplements added, growth tended to be faster at 25 than 37 C. Generally, plasmid-bearing strains grew slower than plasmid-cured strains in most media at 37 C due to virulence plasmid expression retarding growth. In some instances at 37 C, it was observed that the growth rates of both plasmid-bearing and plasmid-cured strains were comparable, indicating the in¯uence of added selective agent/s negating any effects associated with virulence plasmid expression. The effects of selective agents, incubation temperature and virulence plasmid carriage on the growth kinetics of Y. enterocolitica are discussed.

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Incidence and pathogenicity of Yersinia spp. isolates from poultry in Spain

Capita

2002

Food Microbiology

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Simple assay of calcium dependency for virulent plasmid-bearing clones of Yersinia enterocolitica

Cited by 36 publications

References 8 publications

DETECTION OF YERSINIA PESTIS BY COMPARISON OF VIRULENCE PLASMID (PYV/PCD)‐ASSOCIATED PHENOTYPES IN YERSINIA SPECIES*

DETECTION OF YERSINIA PESTIS BY COMPARISON OF VIRULENCE PLASMID (PYV/PCD)‐ASSOCIATED PHENOTYPES IN YERSINIA SPECIES*

The effect of temperature and selective agents on the growth of Yersinia enterocolitica serotype O:3 in pure culture

Incidence and pathogenicity of Yersinia spp. isolates from poultry in Spain

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