Isolation and Characterization of C. jejuni Strains

The three isolates of C. jejuni used in the study were 48 (Penner serotype O:19), 75 (serotype O:3), and 111 (serotype O: in 1,44) which originated from the stools of diarrheic patients as stated above. They were found to colonize mouse intestine in previous studies [11 (link), 22 (link)]. Stock cultures were maintained in Brucella broth (Becton & Dickinson, Sparks, MD) with 15% (vol/vol) glycerol at -70°C. Depending upon the purpose, three types of media were used for cultivation. These were: blood agar base (Oxoid, Basingstoke, Hampshire, England) with 5% defibrinated sheep blood; Campylobacter-selective agar with laked horse blood, growth supplement and selective supplement (Oxoid); and a biphasic medium with brain heart infusion agar and broth supplemented with 1% yeast extract [23 (link)]. Cultures were incubated at 42°C for 48 h in a microaerobic atmosphere generated by Campigen (Oxoid). The identity of the bacteria was confirmed by cultural characteristics and molecular methods. The three serotypes generated unique fingerprints by flaA restriction fragment length polymorphism (RFLP) analysis [24 (link)].

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Albert M.J., Haridas S., Ebenezer M., Raghupathy R, & Khan I. (2015). Immunization with a Double-Mutant (R192G/L211A) of the Heat-Labile Enterotoxin of Escherichia coli Offers Partial Protection against Campylobacter jejuni in an Adult Mouse Intestinal Colonization Model. PLoS ONE, 10(11), e0142090.

Publication 2015

Brucella broth blood agar base Campylobacter-selective agar

Agar Atmosphere Bacteria Blood Brain Brucella Campylobacter Glycerol Heart Horse Intestine Mouse Patients Restriction fragment length polymorphism Sheep Stools Yeast

Corresponding Organization : Kuwait University

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Variable analysis

independent variables

Three isolates of C. jejuni: 48 (Penner serotype O:19), 75 (serotype O:3), and 111 (serotype O:1,44)

dependent variables

Ability of the three C. jejuni isolates to colonize mouse intestine

control variables

Incubation temperature (42°C)
Incubation time (48 h)
Microaerobic atmosphere generated by Campigen (Oxoid)
Culture media (blood agar base with 5% defibrinated sheep blood, Campylobacter-selective agar with laked horse blood, growth supplement and selective supplement, and biphasic medium with brain heart infusion agar and broth supplemented with 1% yeast extract)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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