After C. jejuni infection, the pathogen loads were determined in fecal samples daily, and upon necropsy on day 6 p.i. in luminal samples from the stomach, duodenum, ileum, and colon by culture as described previously [46 (link)]. In brief, intraluminal gastrointestinal samples were homogenized in sterile phosphate-buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA, USA) with a sterile pestle and serial dilutions plated onto karmali agar (Oxoid, Wesel, Germany) and incubated under microaerophilic conditions in a jar at 37 °C for at least 48 h (CampyGas Packs; Oxoid, Wesel, Germany). The detection limit of viable pathogens was 100 CFU per g.
Campygas packs
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The CampyGas Packs are gas packs designed for use in the cultivation of Campylobacter species. They provide the optimal gas environment required for the growth and isolation of these microorganisms in the laboratory.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (4 mentions)
Sterile pbs, by Thermo Fisher Scientific (2 mentions)
Karmali agar, by Thermo Fisher Scientific (1 mentions)
Selective karmali agar plates, by Thermo Fisher Scientific (1 mentions)
Karmali agar plates, by Thermo Fisher Scientific (1 mentions)
4 protocols using campygas packs
1
Quantifying C. jejuni Pathogen Loads
2
Infection of Humanized Mice with C. jejuni
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C. jejuni strain 81-176 bacterial stocks were stored at −80 °C. After thawing, the bacteria were streaked out and incubated on selective karmali agar plates (purchased from Oxoid, Wesel, Germany) at 37 °C for 48 h in a jar under microaerophilic conditions (CampyGas Packs; Oxoid, Wesel, Germany). Grown C. jejuni bacteria were harvested in sterile PBS (Thermo Fisher Scientific, Waltham, MA, USA) immediately before infection. Age- and sex-matched human intestinal microbiota-associated (hma) IL-10−/− mice (3-month-old littermates) were then infected perorally with 109 colony forming units (CFU) of the pathogen (in 0.3 mL) on d0 and d1 (Fig. 1A ) by gavage as reported earlier [48 (link)].
3
Quantification of Campylobacter jejuni in Fecal Samples
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The numbers of viable C. jejuni cells were quantified in daily collected fecal samples (day 2-5 p.i.) and additionally, upon necropsy (day 6 p.i.) from intraluminal gastrointestinal specimens. Samples were homogenized in sterile PBS (Thermo Fisher Scientific, Waltham, MA, USA) using a sterile pestle, and serial dilutions were plated on Karmali agar plates (Oxoid, Wesel, Germany). The inoculated plates were then incubated at 37 8C for at least 48 h under microaerophilic conditions (in a jar containing CampyGas Packs; Oxoid, Wesel, Germany) following previous protocols [26] (link). The limit of detection for viable C. jejuni cells was 100 CFU per g.
4
Quantifying Campylobacter Gastrointestinal Load
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For the determination of gastrointestinal pathogen loads, the numbers of live C. jejuni bacteria were monitored in fecal samples daily post-infection (p.i.), and upon necropsy in intraluminal gastrointestinal samples taken from the stomach, duodenum, ileum, and colon lumen that were subsequently homogenized in PBS (Thermo Fisher Scientific, Waltham, MA, USA). The C. jejuni bacteria were quantified by counting of CFU after growth of serial dilutions of intestinal samples on karmali agar plates placed in a jar for at least 48 h at 37 °C under microaerophilic conditions (CampyGas Packs; Oxoid, Wesel, Germany) as described earlier [48 (link)]. The detection limit of viable pathogens was 100 CFU per g fecal sample.
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