H. pylori strains P1WT and SS1, and H. felis have been reported previously [17 (link)18 (link)]. H. pylori was routinely grown on Campylobacter agar plates or Brucella broth (BD, Sparks, NV, USA) containing 10% of FBS, 10 µg/ mL of vancomycin (Sigma-Aldrich, California, USA), 5 µg/mL of trimethoprim (Sigma), and 1 µg/mL of nystatin (Sigma) at 37℃ under microaerobic conditions using GasPak™ EZ Campy Container System (BD). H. felis was grown on Brucella broth containing 10% of FBS at 37℃ under microaerobic conditions using GasPak™ EZ Campy Container System (BD).
Gaspak ez campy container system
Manufactured by BD
Sourced in United States
The GasPak™ EZ Campy Container System is a laboratory equipment used for the cultivation and growth of Campylobacter species. It provides a controlled atmosphere suitable for the incubation of Campylobacter samples.
Automatically generated - may contain errors
Lab products found in correlation
Brucella broth, by BD (4 mentions)
Brucella broth, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
H pylori selective supplement, by Thermo Fisher Scientific (1 mentions)
Brain heart infusion agar plates, by BD (1 mentions)
2 3 5 triphenyltetrazolium chloride ttc, by Merck Group (1 mentions)
Campylobacter selective agar, by BD (1 mentions)
Campypak plus packs, by BD (1 mentions)
Nystatin, by Merck Group (1 mentions)
Vancomycin, by Merck Group (1 mentions)
Trimethoprim, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Horse serum, by Atlanta Biologicals (1 mentions)
Allegra x 30r, by Beckman Coulter (1 mentions)
Brucella broth medium, by BD (1 mentions)
Etest, by bioMérieux (1 mentions)
Laked horse blood, by Thermo Fisher Scientific (1 mentions)
Bolton enrichment broth, by Thermo Fisher Scientific (1 mentions)
9 protocols using gaspak ez campy container system
1
Culturing H. pylori and H. felis
2
Murine H. felis and H. pylori Infection Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H felis (ATCC 49179) was cultured in sterile-filtered brucella broth (211088; Becton Dickinson, Sparks, MD) containing 10% fetal bovine serum (FBS). Mouse-adapted H pylori Sydney strain 165 (link) was cultured on the Brain Heart Infusion Agar plates (241830; Becton Dickinson) containing 10% sheep blood and H pylori Selective Supplement (SR0147E; Thermo Fisher Scientific, Rockford, IL). The brucella broth and Brain Heart Infusion Agar plates were maintained under microaerobic conditions produced using a GasPak EZ Campy Container System (260680; Becton Dickinson) at 37°C. The brucella broth was maintained at 150 rpm shaking. After 24-hour fasting, 8-week-old mice were orally administered 0.2 mL suspension containing 2 × 108H felis or 1 × 109 colony-forming unit/mL H pylori 4 times every other day.
3
Rapid Campylobacter Detection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following chemicals, drugs, and test kits were used for the experiments: urease test kit (ClO test) was obtained from USA. Brain Heart Infusion (BHI) agar, Brain Heart Infusion (BHI) broth, Skirrow's supplement (SR69) Oxoid England, glycerol, cephalothin (30 μg), and nalidixic acid (30 μg) antibiotic disc were obtained from Mast Group Ltd., Merseyside, UK. GasPak EZ Campy Container System was obtained from Becton Dickinson, USA. Sodium hydroxide, magnesium chloride, sucrose, glucose, methanol (99.8%), and ethanol (99.8%) were obtained from British Drug House, (BDH) (London, UK). 2,3,5-Triphenyltetrazolium chloride (TTC) was obtained from Sigma Aldrich. Expired blood was obtained from the Blood Bank of the Cape Coast Regional hospital, Cape Coast, Ghana.
4
Culturing Helicobacter spp. for Murine Studies
5
Helicobacter felis Infection Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
H. felis (CS1 strain) stocks were stored in 50% glycerol solution at −80°C. Bacteria were cultured in sterile-filtered Brucella broth (BD, Franklin Lakes, NJ) plus 10% FBS (Atlanta Biologicals, Lawrenceville, GA) using the GasPak™ EZ Campy Container System (BD) at 37°C with 150 rpm shaking. The cultures were spun down at 2700 rpm at room temperature, and the pellets resuspended in Brucella broth plus 10% FBS (Thermo Fisher Scientific, Houston, TX). Cells were counted using a hemocytometer by diluting the cells 1:100 in 9:1 HBSS/Formalin solution. Mice were gavaged 3 times over 3 days with 108H. felis cells in 100 μl of Brucella broth. Mice were infected for 6 months prior to euthanasia.
+ Expand
H. felis (CS1 strain) stocks were stored in 50% glycerol solution at −80°C. Bacteria were cultured in sterile-filtered Brucella broth (BD, Franklin Lakes, NJ) plus 10% FBS (Atlanta Biologicals, Lawrenceville, GA) using the GasPak™ EZ Campy Container System (BD) at 37°C with 150 rpm shaking. The cultures were spun down at 2700 rpm at room temperature, and the pellets resuspended in Brucella broth plus 10% FBS (Thermo Fisher Scientific, Houston, TX). Cells were counted using a hemocytometer by diluting the cells 1:100 in 9:1 HBSS/Formalin solution. Mice were gavaged 3 times over 3 days with 108H. felis cells in 100 μl of Brucella broth. Mice were infected for 6 months prior to euthanasia.
6
Helicobacter felis Infection Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H felis (CS1 strain) were cultured in sterile-filtered Brucella broth medium (BD, Franklin Lakes, NJ) plus 10% horse serum (Atlanta Biologicals, Lawrenceville, GA) using the GasPak EZ Campy Container System (BD) at 37°C with shaking at 100 rpm. The bacterial culture suspension was collected by centrifuging at 1400 × g in a Beckman Allegra X-30r centrifuge at room temperature. Subsequently the bacterial pellets were resuspended in saline solution. Optical density of the resuspended H felis was determined and the final concentration was adjusted to 109H felis cells in 1 mL saline. Mice were gavaged once a day with 108H felis CFUs in 100 μL saline for 3 consecutive days. Control mice were gavaged with 100 μL of saline solution.
7
Helicobacter pylori Cultivation and Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This project employed the use of the Pre-mouse Sydney Strain 1 (PMSS1) of Helicobacter pylori or the mouse-passaged derivative, SS1. Bacteria were grown on Trypticase soy agar (TSA) plates containing 5% sheep blood. Alternatively, bacteria were grown in Brucella broth containing 10% heat-inactivated fetal bovine serum (FBS) and 10μg/ml vancomycin. Cultures were grown at 37°C either in an incubator supplemented with 5% CO2 or under microaerophilic conditions generated by a GasPak™ EZ Campy Container System (BD). Liquid cultures for infection were grown under microaerophilic conditions with shaking at 150 rpm. In order to isolate H. pylori from the stomachs of the mice, stomach homogenate was placed on TSA plates containing sheep blood (5%), nalidixic acid (10 μg/ml), vancomycin (50 μg/ml), amphotericin (2 μg/ml) and bacitracin (100 μg/ml) and cultured under microaerophilic conditions at 37°C for 3–5 days.
8
Bacterial Strain Susceptibility Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial strains used in this study are shown in Table 1 . All testing was conducted in Mueller-Hinton broth or agar supplemented with 50 μg/ml of calcium and 12.5 μg/ml of magnesium (CA-SMHB and CA-SMHA, respectively) as required for DAP (45 , 46 ), unless otherwise stated. Cultures were incubated at 37°C in a GasPak EZ Campy container system (BD) unless otherwise stated. Etest (bioMérieux, Inc.) susceptibilities were determined on tryptic soy agar (TSA) supplemented with 5% defibrinated horse blood (Remel) or on MHA.
9
Isolation of Campylobacter from Water
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The presence of Campylobacter in the water samples was investigated following the method described by Walters et al. (2007) . Briefly, 200 ml of the water sample was filtered through a 0.45 µm Millipore membrane and the filter was placed in 30 ml of Bolton Enrichment Broth (Oxoid) containing selective supplement (Oxoid SR183E) and 5% (v/v) laked horse blood (Oxoid). The broth cultures were incubated under microaerophilic conditions (5-15% oxygen) using a GasPak EZ Campy container system (BD Diagnostic Systems, New Jersey), at The enriched cultures were also grown on Blood-free Campylobacter selective agar (Oxoid) followed by incubation at 42 o C under microaerophilic conditions as described above for 48 hours. Presumptive Campylobacter colonies were identified based on colony morphology, i.e. grey and moist colonies.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!