Chapter X Methods in Campylobacter

Ullmann, U.

doi:10.1016/s0580-9517(08)70380-5

Cited by 4 publications

(1 citation statement)

References 23 publications

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“…The C. jejuni isolates were subcultured on blood agar base (Oxoid) supplemented with 5 % (v/v) defibrinated sheep blood (Gibco) and Blaser-Wang selective supplement (Oxoid) and incubated at 42 uC in anaerobic jars under a gas atmosphere of 5 % O 2 , 15 % CO 2 and 80 % N 2 (BOC). The isolates were identified by standard tests including colonial morphology, Gram stain, motility, positive oxidase and catalase tests, lack of growth at 25 uC and positive hippurate test (Ullmann, 1979). After primary isolation, the isolates were subcultured only once to minimize cultural changes, after which samples of each, stored at 270 uC in Brucella broth (Oxoid) and glycerol (60 : 40 v/v), were used for a limit of five subcultures before a new stored culture was used.…”

Section: Methodsmentioning

confidence: 99%

Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources

Coote

Stewart‐Tull

Owen³

et al. 2007

Journal of Medical Microbiology

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Campylobacter jejuni is a major cause of human diarrhoeal disease, but specific virulence mechanisms have not been well defined. This blinded study was undertaken with 40 C. jejuni isolates from different sources to determine their haemolytic, cytotoxic and adhesion and invasion activities towards mammalian cells. The results were correlated with source of isolation and genetic makeup by amplified fragment length polymorphism (AFLP) typing. The isolates had variable degrees of haemolytic activity against rabbit erythrocytes and cytotoxicity towards CaCo-2, HeLa and Vero cells. The data indicated that the haemolytic and cytotoxic activities were due to separate factors. A range of cytotoxicity was exhibited, whereby some strains had no activity against the target cells and others had activity against all three cell lines. Certain strains had activity against CaCo-2 cells but little or no activity against the other cells, while others exhibited the opposite phenotype. The data suggested that the cytotoxicity assay with the different cell lines may have detected more than one cytotoxin. A wide variation between isolates was observed for both adherence and invasion with all three cell lines, yet, overall, the strains showed a significantly greater invasion capacity for CaCo-2. There was no clear relationship between source of isolation or disease manifestation and possession of statistically significantly higher levels of particular virulence-associated factors although, in some cases, a correlation between cytotoxicity and cell invasion was evident. Five AFLP clusters, each representing two to eleven isolates with similar profiles, were observed at the 90 % similarity level. Some AFLP groups contained isolates with a common serotype, but each group had C. jejuni isolates from more than one source with the exception of group IV, which contained only human isolates. Isolates with high cytotoxic activity against CaCo-2 cells were confined to groups I, III and IV and a group of unrelated strains (U). Group II isolates had uniformly low cytotoxicity. Isolates in groups I, V and U were more invasive for CaCo-2 cells than isolates in groups II, III and IV. The strain differences in cytotoxicity or invasion did not correlate with source of isolation.

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Section: Methodsmentioning

confidence: 99%

Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources

Coote

Stewart‐Tull

Owen³

et al. 2007

Journal of Medical Microbiology

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A rapid, colourimetric assay for cytotoxin activity inCampylobacter jejuni

Coote

Arain

1996

FEMS Immunol Med Microbiol

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Cell extracts and culture supernates of Campylobacter jejuni NCTC 11168 and three isolates from faecal samples from patients with enteritis were tested for cytotoxic activity on HeLa and Vero cells using a sensitive and rapid dye reduction assay which represents a simple assay for cytotoxin activity that can be assessed visually or spectrophotometrically in the wells of microplates. The assay was as sensitive as trypan blue exclusion and did not require the use of radioisotopes. A low level of cytotoxin activity, compared to that produced by a control verotoxin 2-producing Escherichia coli strain, was detected in cell extracts of all four strains, but no activity was detected in culture supernates. Production of an enterotoxin was evaluated by reverse passive latex agglutination with anti-cholera toxin antibody, a procedure which also represents a rapid and simple assay for this toxin. No enterotoxin activity was detected in cell extracts or culture supernates from any of the isolates.

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Prevalence of Campylobacter jejuni in two California chicken processing plants

Wempe¹,

Genigeorgis²,

Farver³

et al. 1983

Appl Environ Microbiol

127

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Two federally inspected California chicken processing plants participated in Campylobacter jejuni prevalence studies. Twelve sampling sites were included in each of four groups. Groups were based on bird age, scald water temperature, and plant sampled. Scald water temperatures of 60 degrees C (140 degrees F) did not contribute to a lower prevalence of C. jejuni in edible parts, as did temperatures of 53 degrees C (127 degrees F) and 49 degrees C (120 degrees F). The feather picker and chilling tank were areas of major cross-contamination. C. jejuni was isolated from 68% of the ready-for-market products. The organism was recovered from 60 to 100% of the ceca in the four groups, and some numbers in the fecal material exceeded 10(6)/g. The level of C. jejuni in intestinal tracts seemed to correlate with the presence of the organism in the edible parts.

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Chapter X Methods in Campylobacter

Cited by 4 publications

References 23 publications

Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources

Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources

A rapid, colourimetric assay for cytotoxin activity inCampylobacter jejuni

Prevalence of Campylobacter jejuni in two California chicken processing plants

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