A set of 30 Gram-negative and 17 Gram-positive strains belonging to five genera of bacteria from the Infectious Diseases Unit Collection of the University of León in Spain were selected for the assessment (Table 1 ). Tryptic soy agar (Scharlau) and Mueller-Hinton broth (Cultimed) as solid and liquid media, respectively, were used to grow E. coli, Salmonella enterica ssp. enterica, and S. aureus; Columbia agar with sheep blood (Oxoid) and brain heart infusion broth (Merck) supplemented with 0.4% yeast extract for the growth of Campylobacter spp.; and fastidious anaerobe agar (Amersham) and brain heart infusion broth for the culture of Clostridium spp. Bacteria were grown in both solid and liquid media for 24 h at 37°C with the exceptions of Campylobacter spp. and Clostridium spp. that were maintained for 48 h. Liquid cultures were grown on an orbital shaker at 60 rpm. Microaerophilic conditions for the culture of Campylobacter spp. were achieved using a jar with a pack of CampyGenTM (Oxoid), whereas anaerobic bacteria were incubated on solid medium in an anaerobic chamber (80% N2, 10% H2, and 10% CO2), or in a jar with a pack of AnaeroGenTM (Oxoid) when liquid medium was used. The bacteria were always resuscitated on solid medium plates to check purity before each assay and all the experiments described below were carried out in triplicate.
Campygentm
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The CampyGenTM is a lab equipment product designed for the detection and identification of Campylobacter species in various sample types. It provides a reliable and efficient solution for microbiological analysis.
Automatically generated - may contain errors
Lab products found in correlation
Tryptic soy agar, by Scharlab (1 mentions)
Brain heart infusion broth, by Merck Group (1 mentions)
Anaerogentm, by Thermo Fisher Scientific (1 mentions)
Columbia agar with sheep blood, by Thermo Fisher Scientific (1 mentions)
Mrs agar, by Thermo Fisher Scientific (1 mentions)
Modified charcoal cefoperazone deoxycholate agar, by Thermo Fisher Scientific (1 mentions)
Karmali agar, by Thermo Fisher Scientific (1 mentions)
4 protocols using campygentm
1
Growth Conditions for Diverse Bacterial Strains
2
Isolation and Preservation of Lactobacilli from Bovine Feces
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1g of cattle fecal samples were added into 9 ml of MRS broth and incubated at 37oC under microaerophilic condition (CampyGenTM Oxoid, UK) for 24 hours, the culture were appropriately plated out on MRS agar (Oxoid, UK) and viable cells were counted. Distinct morphologically different colonies were picked from each plates and sub-cultured to obtain pure cultures. Gram positive and catalase negative isolates were preserved in 50% glycerol stock at -800C.
3
Campylobacter Detection in Animal Faeces
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Faecal samples from each animal were obtained from the rectum and transferred to 100 ml plastic jars. A new and clean pair of gloves were used for each sample and the jars were filled to a maximum of two‐thirds, in order to decrease the risk of the lid opening during transport. The samples were transported chilled to the National Veterinary Institute (SVA, Uppsala, Sweden). All packages reached the laboratory within 48 h. The samples were analysed by direct culture according to ISO 10272: Part 1C (2017). In brief, faecal contents were spread on modified charcoal‐cefoperazone‐deoxycholate agar (mCCDA) (Oxoid, Basingstoke, UK) and the plates were incubated at 37·0°C for 44 ± 4 h in microaerobic atmosphere generated by the use of CampyGenTM (Oxoid). Identification of Campylobacter spp. was based on typical morphological aspects, white to grey colonies with metallic sheen and phase‐contrast microscopic observation with corkscrew movement according to ISO 10272: part 1 (2017).
4
Isolation and Identification of Campylobacter
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For the isolation of the Campylobacter isolates, approximately one gram of each faecal sample was mixed in a sterile container using a sterile swab stick. The excess stool samples were wiped off and the sticks were swabbed onto a (pre-dried) Karmali agar (Oxoid) plate. Plates were incubated at 42°C for 48 hrs in an anaerobic jar with a gas-generating sachet (Oxoid-CampyGenTM) to produce micro aerophilic atmospheric conditions to enable the growth of Campylobacter spp. [23 (link)]. After 48 hrs incubation, smooth, flat, colourless translucent to grey colonies of approximately 1 mm on Karmali agar plates were selected. Since the colonies were often mixed with other bacteria on the Karmali plate, half of the suspected colony was tested for motility using a phase contrast microscope. Colonies showing positive motility were then purified by plating the remaining colonies on 5% horse blood agar plates and Karmali plate. The plates were then incubated for 48 hrs in an anaerobic jar with a gas-generating sachet to produce microaerophilic conditions. Gram-negative, catalase and oxidase positive isolates were stored snap-frozen in 30% glycerol broth at 70°C for further molecular identification (using polymerase chain reaction), antibiotic susceptibility testing and resistance genes determination.
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