scholarly journals Differences in virulence attributes between cytolethal distending toxin positive and negative Campylobacter jejuni strains

2008 ◽  
Vol 57 (3) ◽  
pp. 267-272 ◽  
Author(s):  
Deepika Jain ◽  
Kashi Nath Prasad ◽  
Sushmita Sinha ◽  
Nuzhat Husain
Keyword(s):  
Gastrointestinal Tract ◽  
Campylobacter Jejuni ◽  
Hela Cells ◽  
Histopathological Examination ◽  
Bacterial Pathogen ◽  
Biological Properties ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Culture Supernatants ◽  
Major Target

Campylobacter jejuni is a common gastrointestinal bacterial pathogen. Although cytolethal distending toxin (CDT) is proposed to be an important virulence determinant of this pathogen, how CDT+ and CDT− strains differ in their biological properties remains largely unknown. The virulence properties of CDT+ and CDT− strains were studied on HeLa cells and in the suckling mouse model. Presence of the cdtB gene in Campylobacter species was determined by PCR. Five each of CDT+ and CDT− C. jejuni strains were subjected to adherence, invasion and cytotoxicity assay on the HeLa cell line. Bacterial culture supernatants with and without CDT activity were inoculated intragastrically into 2-day-old suckling mice. The mice were sacrificed within 48 h. Histopathological examination of stomach, jejunum, ileum and colon was performed by haematoxylin/eosin staining. cdtB was detected in 88 % and 14 % of C. jejuni and Campylobacter coli strains, respectively. CDT+ C. jejuni strains adhered to and invaded HeLa cells in significantly higher numbers than CDT− strains [CDT+ vs CDT−, adherence 2.7×104±3.5×104 vs 2.7×102±1.9×102; invasion 1.0×103±1.3×103 vs1.4×101±3.1×101; P<0.01]. Culture supernatants of all CDT+ strains demonstrated CDT activity on HeLa cells. Mice inoculated with supernatant containing CDT activity had moderate to severe pathology in different parts of their gastrointestinal tract, with the colon being the major target. Mice inoculated with supernatant lacking CDT activity showed no significant pathology in the gastrointestinal tract. The results demonstrate that CDT+ C. jejuni strains adhere to and invade epithelial cells more efficiently than CDT− strains. CDT is responsible for intestinal pathology and the colon is the major target.

scholarly journals Campylobacter jejuni Cytolethal Distending Toxin Mediates Release of Interleukin-8 from Intestinal Epithelial Cells

2000 ◽  
Vol 68 (12) ◽  
pp. 6535-6541 ◽  
Author(s):  
Thomas E. Hickey ◽  
Annette L. McVeigh ◽  
Daniel A. Scott ◽  
Ronda E. Michielutti ◽  
Alyssa Bixby ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Intestinal Epithelial Cells ◽  
Sodium Deoxycholate ◽  
Interleukin 8 ◽  
Live Cells ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Intestinal Epithelial ◽  
Protein Subunits ◽  
Shuttle Plasmid

ABSTRACT Live cells of Campylobacter jejuni andCampylobacter coli can induce release of interleukin-8 (IL-8) from INT407 cells. Additionally, membrane fractions of C. jejuni 81-176, but not membrane fractions of C. colistrains, can also induce release of IL-8. Membrane preparations from 81-176 mutants defective in any of the three membrane-associated protein subunits of cytolethal distending toxin (CDT) were unable to induce IL-8. The presence of the three cdt genes on a shuttle plasmid in trans restored both CDT activity and the ability to release IL-8 to membrane fractions. However, CDT mutations did not affect the ability of 81-176 to induce IL-8 during adherence to or invasion of INT407 cells. When C. jejuni cdt genes were transferred on a shuttle plasmid into a C. coli strain lacking CDT, membrane preparations became positive in both CDT and IL-8 assays. Growth of C. jejuni in physiological levels of sodium deoxycholate released all three CDT proteins, as well as CDT activity and IL-8 activity, from membranes into supernatants. Antibodies against recombinant forms of each of the three CDT subunit proteins neutralized both CDT activity and the activity responsible for IL-8 release. The data suggest that C. jejuni can induce IL-8 release from INT407 cells by two independent mechanisms, one of which requires adherence and/or invasion and the second of which requires CDT.

scholarly journals A PCR-RFLP assay for the detection and differentiation of Campylobacter jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis

2014 ◽  
Vol 63 (5) ◽  
pp. 659-666 ◽  
Author(s):  
Kazumasa Kamei ◽  
Masahiro Asakura ◽  
Srinuan Somroop ◽  
Noritoshi Hatanaka ◽  
Atsushi Hinenoya ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Sensitivity And Specificity ◽  
Gastrointestinal Diseases ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Restriction Digestion ◽  
Animal Diseases ◽  
Pcr Rflp ◽  
Rflp Assay ◽  
Human And Animal Diseases

Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.

scholarly journals Monitoring of Selected Genes in Campylobacter Jejuni and Campylobacter Coli Isolates from Domestic Animals

2012 ◽  
Vol 56 (3) ◽  
pp. 283-286
Author(s):  
Marek Selwet ◽  
Mariola Galbas
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Domestic Animals ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Frequency Of Occurrence

Abstract In the present study, the presence of flaA, cadF, cdtB, and iam genes of Campylobacter sp. were analysed using PCR. Material for analyses comprised 100 Campylobacter sp. isolates obtained from healthy broiler chickens, fatteners, and calves, among which 84 isolates were ascribed to Campylobacter jejeuni and 16 to Campylobacter coli. All isolates (100%) had the cadF gene responsible for adhesion and the flaA gene determining the motility of the analysed bacteria. The frequency of occurrence of the cdtB gene responsible for the production of the cytolethal distending toxin (CDT) was determined to be high (98.6% in broiler chickens, 75% in fatteners, 62.5% in calves). In case of the iam gene, the highest frequency was recorded in Campylobacter sp. isolated from broiler chickens (84.7%), while in strains collected from fatteners and calves it was lower, amounting to 41.7% and 18.8%, respectively.

Effect of iron concentration on toxin production in Campylobacter jejuni and Campylobacter coli

1986 ◽  
Vol 32 (5) ◽  
pp. 395-401 ◽  
Author(s):  
Barbara A. McCardell ◽  
Joseph M. Madden ◽  
John T. Stanfield
Keyword(s):  
Campylobacter Jejuni ◽  
Ferrous Sulfate ◽  
Culture Media ◽  
Iron Concentration ◽  
Toxin Production ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Casamino Acids ◽  
Culture Supernatants ◽  
Campylobacter Spp

The effect of iron concentrations in culture media on supernatant yields of Campylobacter cytotonic toxin (CCT) was studied. Of the 118 Campylobacter spp. strains surveyed, 78.8% produced toxin in brucella broth or in casamino acids – yeast extract (CYE) broth. When the iron concentration of CYE was increased from 0.44 μg/mL (7.9 μM) to 0.65 μg/mL (11.6 μM) by the addition of ferric chloride, 94.9% of the strains were positive for toxin in a ganglioside GM1 based, enzyme-linked immunosorbent assay, using antibody to affinity-purified CCT. The addition of iron as ferrous sulfate was less effective. When four toxin-positive strains were grown in a deferrated medium of conalbumin-treated CYE with 0.04–0.08 μg iron/mL (0.72–1.43 μM), two of the culture supernatants became negative (absorbance at 410 nm, < 0.1 and < 10 ng CCT/mL), and two produced about 90% less CCT but were still classified as positive (absorbance, ≥ 0.1 and ≥ 10 ng CCT/mL). It was therefore concluded that the production of CCT by Campylobacter spp. is influenced by iron concentration.

scholarly journals Campylobacteriosis and other bacterial gastrointestinal diseases in Sofia, Bulgaria for the period 1987 – 2008

2010 ◽  
Vol 15 (4) ◽  
Author(s):  
K Ivanova ◽  
M Мarinа ◽  
P Petrov ◽  
T Kantardjiev
Keyword(s):  
Escherichia Coli ◽  
Campylobacter Jejuni ◽  
Bacterial Pathogen ◽  
Gastrointestinal Diseases ◽  
Campylobacter Coli ◽  
Systematic Data ◽  
Thermophilic Campylobacter ◽  
Diarrhoeal Diseases ◽  
Campylobacter Infection ◽  
The City

Campylobacter is the most commonly reported enteric bacterial pathogen in humans. We still do not have any systematic data concerning campylobacteriosis in Bulgaria. For that reason, we present data of the thermophilic Campylobacter jejuni and Campylobacter coli in the aetiology of diarrhoeal diseases in Sofia, for the period from 1987 to 2008. The study included patients from 0 to over 65 years-old. A total of 51,607 faecal specimens were screened for Campylobacter. C. jejuni and C. coli were detected in 3.58% (1,847) of the strains, with the highest percentage in 1988 (7.5%) and the lowest in 2006 (0.3%). Campylobacteriosis occurred most frequently in the wet months of March, April, May and June, with 105, 102, 124 and 141 cases, respectively, and was rare in January with 25 cases. The most affected groups were children between 0 and 4 years of age (52%) and between five and 14 years of age (30%). Campylobacter infection occurred in 22% of all bacterial gastrointestinal diseases in the city of Sofia during the study period. Salmonella was the most frequently identified pathogen with 32%, followed by Shigella (30%), Campylobacter (22%) and diarrhoeagenic Escherichia coli (16%). The study shows that Campylobacter plays an important role as a bacterial cause of enterocolitis in Sofia, Bulgaria.

scholarly journals Association with HeLa cells of Campylobacter jejuni and Campylobacter coli isolated from human feces.

1986 ◽  
Vol 54 (2) ◽  
pp. 283-287 ◽  
Author(s):  
J L Fauchere ◽  
A Rosenau ◽  
M Veron ◽  
E N Moyen ◽  
S Richard ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Hela Cells ◽  
Campylobacter Coli ◽  
Human Feces

scholarly journals Detection of Cytolethal Distending Toxin Activity and cdt Genes in Campylobacter spp. Isolated from Chicken Carcasses

1999 ◽  
Vol 65 (4) ◽  
pp. 1501-1505 ◽  
Author(s):  
Aysegul Eyigor ◽  
Karl A. Dawson ◽  
Bruce E. Langlois ◽  
Carol L. Pickett
Keyword(s):  
Campylobacter Jejuni ◽  
Restriction Analysis ◽  
Primary Source ◽  
High Rate ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Pcr Products ◽  
Species Specific ◽  
Human Infections ◽  
Campylobacter Spp

ABSTRACT This study was designed to determine whether isolates from chicken carcasses, the primary source of Campylobacter jejuni andCampylobacter coli in human infections, commonly carry thecdt genes and also whether active cytolethal distending toxin (CDT) is produced by these isolates. Campylobacterspp. were isolated from all 91 fresh chicken carcasses purchased from local supermarkets. Campylobacter spp. were identified on the basis of both biochemical and PCR tests. Of the 105 isolates, 70 (67%) were identified as C. jejuni, and 35 (33%) were identified as C. coli. PCR tests amplified portions of thecdt genes from all 105 isolates. Restriction analysis of PCR products indicated that there appeared to be species-specific differences between the C. jejuni and C. coli cdt genes, but that the restriction patterns of thecdt genes within strains of the same species were almost invariant. Quantitation of active CDT levels produced by the isolates indicated that all C. jejuni strains except four (94%) had mean CDT titers greater than 100. Only one C. jejuni strain appeared to produce no active CDT. C. coli isolates produced little or no toxin. These results confirm the high rate ofCampylobacter sp. contamination of fresh chicken carcasses and indicate that cdt genes may be universally present inC. jejuni and C. coli isolates from chicken carcasses.

Adherence, Invasion, Toxigenic, and Chemotactic Properties of Mexican Campylobacter Strains

2010 ◽  
Vol 73 (11) ◽  
pp. 2093-2098 ◽  
Author(s):  
ISMAEL MALAGÓN ◽  
SANTOS GARCÍA ◽  
NORMA HEREDIA
Keyword(s):  
Public Health ◽  
Campylobacter Jejuni ◽  
Vero Cells ◽  
Campylobacter Coli ◽  
Cytolethal Distending Toxin ◽  
Wild Type ◽  
Mueller Hinton ◽  
Mueller Hinton Agar ◽  
The Difference ◽  
Type Strains

To determine the virulence factors of Mexican wild-type strains of Campylobacter jejuni and Campylobacter coli, 31 wild-type strains were isolated from food and from humans. The production of cytolethal distending toxin and the adherence and invasion capabilities of these strains were assayed in Vero cells. Hard agar plugs with repellents and attractants were used to examine chemotaxis. Mueller-Hinton agar with supplements was used for motility analysis and to measure hemolytic activity. Nine strains of C. jejuni and eight strains of C. coli exhibited motility, most within a diameter of 2 to 13 mm. Most of the strains reacted to the repellent compounds analyzed, and α- and β-like hemolysis and cytotoxicity in Vero cells were observed for all strains. Isolates adhered to and invaded Vero cells to various degrees. Although strains of C. jejuni exhibited stronger adherence but less invasion compared with strains of C. coli, the difference was not significant (P &gt; 0.05). The strains of C. jejuni and C. coli isolated from food and from patients in Mexico could have major impacts on public health.

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