Bbl gaspak jars

For selective pre-enrichment, 25 g of each sample (chicken meat, liver, or gizzard), were added to 225 mL of a Brucella broth supplemented with 5% sheep defibrinated blood, and DENT selective supplement (Oxoid, Hampshire, UK), and the mixture was homogenized in Stomacher^® 400 (Seward, Worthing, UK). The mixture was then divided into two 250 mL flasks and incubated at 37 °C for 48 h under a microaerophilic condition using BBL GasPak™ jars (Becton Dickinson, Franklin Lakes, NJ, USA), supplemented with CampyGen bags (Oxoid, Hampshire, UK). After incubation, 100 µL of the mixture were inoculated onto Columbia blood agar base (Oxoid, Hampshire, UK), supplemented with 5% sheep blood and DENT selective supplements. The plates were incubated for up to 7 days at 37 °C under a microaerophilic condition, as previously described. Suspected colonies were further identified using Gram’s staining (Gram-negative for Helicobacter spp.), oxidase (positive for Helicobacter spp.), urease (positive for H. pylori), and nitrate reduction (positive for H. pullorum) tests.

For the isolation of H. pylori, 25 g of each sample was suspended in 225 mL of Brucella broth supplemented with 5% sheep defibrinated blood and DENT selective supplement (Oxoid, Hampshire, UK). After incubation at 37 °C for 48 h under a microaerophilic condition using BBL GasPak™ jars (Becton Dickinson, Franklin Lakes, NJ, USA), supplemented with CampyGen bags (Oxoid, Hampshire, UK) 100 µL of the mixture was inoculated onto Columbia blood agar base (Oxoid, Hampshire, UK), supplemented with 5% sheep blood and DENT selective supplements, and then incubated for 5–7 days under a microaerophilic condition. Suspected colonies were further identified using Gram staining (Gram-negative for Helicobacter spp.), oxidase (positive for Helicobacter spp.), urease (positive for H. pylori), and nitrate reduction (positive for H. pullorum) tests.