Assay of crystal violet binding for rapid identification of virulent plasmid-bearing clones of Yersinia enterocolitica

Bhaduri, Saumya; Conway, L K; Lachica, R. V. F.

doi:10.1128/jcm.25.6.1039-1042.1987

Cited by 80 publications

(52 citation statements)

References 22 publications

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“…Of special interest is the use of dyes for differentiation of strains of Y . pestis (Surgalla and Beesley 1969) and Y. entercolitica (Robins-Browne and Prpic 1985;Bhaduri et al 1987;Riley and Toma 1989), in which virulent plasmid-bearing clones acquired a darker colour, due to a differential binding capacity for the dye compared with the plasmid-less avirulent clones. Our results show that Y. ruckeri HSF' and HSFstrains were differentiated by the addition of Congo red to the media although, in this case, HSF -(avirulent) colonies appeared a darker colour than HSF' (virulent) ones.…”

Section: Discussionmentioning

confidence: 99%

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

Furones

Gilpin

Munn

1993

Journal of Applied Bacteriology

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Strains of the bacterial fish pathogen Yersinia ruckeri were identified with the API 20E system and distinguished on the basis of whole cell agglutination with antisera, sorbitol fermentation and polymyxin B sensitivity. Strains which were shown to possess the virulence-associated heat-sensitive factor (HSF) were shown to grow preferentially on culture media containing sodium dodecyl sulphate (SDS) and to produce a creamy deposit around the colonies. By contrast, strains lacking this factor (HSF-) grew poorly and without forming a deposit. Enhancement of the differentiation between the two types was shown by the incorporation of Coomassie brilliant blue dye into agar containing 1% SDS, and the uptake of Coomassie blue and Congo red was shown to be temperature-dependent. Most strains tested were shown to belong to serotype I, and were sensitive to polymyxin and did not ferment sorbitol. With the medium developed most serotype I strains but not those of other serotypes were shown to possess HSF. It is suggested that the medium is used in epidemiological studies of Y. ruckeri.

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Section: Discussionmentioning

confidence: 99%

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

Furones

Gilpin

Munn

1993

Journal of Applied Bacteriology

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“…On calcium adequate BHA the colony size at 37C (1.13 mm) and morphology of JD193 and JD217 were similar to that of GER as reported previously (Bhaduri et al 1990). Both JD 193 and JD217 also displayed other plasmid-associated phenotypes at 37C including Lcr, CV binding, AA and HP as observed with GER (Bhaduri et al 1987;Bhaduri et al 1990) (Table 2). Mouse virulence assays were positive for GER and JD193 but not for JD217 as shown by oral infection which led to diarrhea from 5 to 7 days of post infection in iron-overloaded mice ( Table 2).…”

Section: Resultsmentioning

confidence: 53%

“…Detailed descriptions of these assay conditions are given elsewhere (Bhaduri et al 1987;Bhaduri et al 1990). AA was determined as previously described (Bhaduri et al 1987) with Eagle's minimal essential medium supplemented with 10% fetal bovine serum. HP was examined by latex particle agglutination (LPA) test (Bhaduri et ul.…”

Section: Assays Of Plasmid-associated Phenotypic Characteristicsmentioning

confidence: 99%

“…BHADURI phenotypic characteristics expressed at 37C include colony morphology (appearance as small colony of 1.13 mm in diameter due to slower growth rate), calcium-dependency (also known as low-calcium response: Lcr; appearance of pinpoint colony diameter of 0.36 mm in low calcium medium), crystal violet (CV) binding, autoagglutination (AA), hydrophobicity (HP), serum resistance, and detachment of cells in culture. These characteristics have been used to distinguish between plasmid-bearing virulent and plasmidless avirulent strains and also have been correlated with the pathogenicity of the organism viz the mouse virulence test (Bhaduri 1990a;Bhaduri et al 1987;Doyle and Cliver 1990;Kapperud 1991;Kwaga and Iversen 1991;Lazere and Gemski 1983;Mazigh et al 1983;Portnoy and Martinez 1985;Robins-Browne et al 1989); though the mechanisms involved invirulence remain unclear. Studies by several investigators (Bissett et al 1990;Kwaga and Iversen 1991;Mazigh et al 1985;Morris et al 1991;Noble et al 1987;Prpic et al 1985) suggest that the presence of the virulence plasmid and its expression in Y. enterocolitica may not unequivocally indicate pathogenicity.…”

Section: Introductionmentioning

confidence: 99%

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LACK OF CORRELATION BETWEEN PLASMID‐ASSOCIATED PHENOTYPES OF YERSINIA ENTEROCOLITICA AND PATHOGENICITY IN THE MOUSE¹

Bhaduri

1996

Journal of Food Safety

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Various phenotypic characteristics have been correlated with the pathogenicity of plasmid‐bearing virulent strains of Yersinia enterocolitica. Two transposon Tn801‐insertion derivatives (JD193 and JD217) of the virulence plasmid from serotype 0:3 were used to determine the correlation between pathogenicity and plasmid‐associated properties of this organism. Both Tn801‐inserted derivatives expressed five plasmid‐associated phenotypic characteristics at 37C: (1) colony morphology, (2) calcium‐dependent growth or low‐calcium response, (3) crystal violet binding, (4) autoagglutination, and (5) hydrophobicity. However, for mouse pathogenicity only JD193 was positive whereas, JD217 was avirulent for mice. Thus, it is possible to have a lack of correlation between plasmid‐mediated traits and the actual pathogenicity of the organism in the mouse; however, these plasmid‐mediated phenotypic characteristics provide simple and efficient techniques to evaluate the virulence potential of wild‐type strains isolated from food poisoning outbreaks and clinical cases.

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“…The cells were diluted to 10 3 cells per mL (determined by A 600 ) in 0.1% peptone water and 100 mL surface plated onto BHA. After 24-48 h of incubation at 37C, the CV (Becton-Dickinson) binding assay was performed by gently flooding the plates with 8 mL of a 100-mg/mL solution of CV for 2 min and then the CV was decanted (Bhaduri et al 1987). The binding of CV to P + colonies was observed by their darkviolet appearance, while Pcolonies failed to bind the dye and remained white.…”

Section: Determination Of Pyv/pcd-associated Phenotypic Characteristicsmentioning

confidence: 99%

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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Assay of crystal violet binding for rapid identification of virulent plasmid-bearing clones of Yersinia enterocolitica

Cited by 80 publications

References 22 publications

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

LACK OF CORRELATION BETWEEN PLASMID‐ASSOCIATED PHENOTYPES OF YERSINIA ENTEROCOLITICA AND PATHOGENICITY IN THE MOUSE¹

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

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Assay of crystal violet binding for rapid identification of virulent plasmid-bearing clones of Yersinia enterocolitica

Cited by 80 publications

References 22 publications

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor

LACK OF CORRELATION BETWEEN PLASMID‐ASSOCIATED PHENOTYPES OF YERSINIA ENTEROCOLITICA AND PATHOGENICITY IN THE MOUSE1

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

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LACK OF CORRELATION BETWEEN PLASMID‐ASSOCIATED PHENOTYPES OF YERSINIA ENTEROCOLITICA AND PATHOGENICITY IN THE MOUSE¹