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.
Campygen bags
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
CampyGen bags are a type of laboratory equipment used to create a microaerophilic atmosphere, which is essential for the growth and cultivation of Campylobacter species. These bags provide a controlled environment with a specific gas composition, typically containing a mixture of oxygen, carbon dioxide, and nitrogen, to support the growth of these fastidious microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Dent selective supplement, by Thermo Fisher Scientific (4 mentions)
Columbia blood agar base, by Thermo Fisher Scientific (2 mentions)
Bbl gaspak jars, by BD (2 mentions)
Stomacher 400, by Seward Medical (1 mentions)
Modified charcoal cefoperazone deoxycholate agar plates, by Thermo Fisher Scientific (1 mentions)
4 protocols using campygen bags
1
Selective Enrichment and Identification of Helicobacter
2
Isolation and Identification of H. pylori
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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.
3
Multidrug-resistant bacteria co-culture
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P. aeruginosa ATCC 12453 and E. coli ATCC 25922 were used to co-culture with C. jejuni ATCC 33560. C.jejuni was cultured on Melluler-Hinton agar (Oxoid, Thermo Fisher, Hampshire, UK) under microaerophilic conditions (5% N2, 10% CO2, and 85% O2) using CampyGen bags (Oxoid, Hampshire, UK) at 42 °C for 48 h. E.coli and P. aeruginosa were also cultured on the same medium under aerobic conditions and incubated at 37 °C for 24 h (Karruli et al., 2023 (link), Suwono et al., 2021 (link)). A single colony from each bacterial culture was sub-cultured in Melluler-Hinton broth (Oxoid, Thermo Fisher, Hampshire, UK) and kept under the desired conditions for growth, then an equal volume was extracted from each broth to attain an absorbance of 0.063 at 600 nm, which was then used for co-culture purposes.
4
Isolation of Campylobacter from Milk
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One hundred unpasteurized milk samples were collected: 50 samples from goats, and 50 samples from cows. Sample processing started with adjusting the pH of milk to 7.6, followed by centrifugation at 12,000× g for 4 min. The supernatant was discarded, and the pellet was suspended in 1 mL pre-enriched Bolton broth, then the volume was completed to 10 mL by Bolton broth. Each sample was subjected to pre-enrichment and enrichment steps under microaerobic conditions (5% N2, 10% CO2, and 85% O2) using CampyGen bags (Oxoid, Hampshire, UK). A loopful of the mixture was streaked on modified charcoal cefoperazone deoxycholate agar (mCCDA) plates (Oxoid, Hampshire, UK) and incubated under microaerobic conditions at 42 °C for 48 h.
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