An Intragastric Gavage Technique for Controlled Helicobacter Infection in Mice
Published: September 29, 2023
Abstract
Source: D'Costa, K. et al., Mouse Models Of Helicobacter Infection And Gastric Pathologies. J. Vis. Exp. (2018)
This video demonstrates the establishment of Helicobacter pylori infection through the intragastric gavage technique in a mouse model. The bacterial suspension is administered via a catheter inserted through the mouth into the stomach. Inoculated bacteria colonize and establish an infection in the stomach, which can be used to study the host-pathogen interaction.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Growth and Preparation of Bacterial Inocula Thaw glycerol stocks of Helicobacter pylori (H. pylori) or H. felis from -80 °C and subculture on horse blood agar (HBA) plates comprising: Blood Agar Base No.2 (see Table of Materials); a modified "Skirrow's antibiotic s…
Representative Results
Figure 1: Image demonstrating the oral gavage technique. A disposable 1 mL syringe and flexible catheter are used to deliver ≥105 CFU of bacterial inocula to a mouse via the intragastric route. The mouse was anesthetized using methoxyflurane and held in a firm grip at the neck, allowing for access of the catheter to the stomach via the esophagus.
Divulgazioni
The authors have nothing to disclose.
Materials
| Bacteriological reagents | |||
| Oxoid Blood Agar Base No.2 | Thermo Fischer Scientific | CM0271B | Dissolve in deinonized water prior to sterilization |
| Premium Defibrinated Horse blood | Australian Ethical Biologicals | PDHB100 | |
| Bacto Brain Heart Infusion Broth | BD Bioscience | 237500 | Dissolve in deinonized water prior to sterilization |
| CampyGen gas packs | Thermo Fischer Scientific | CN0035A/CN0025A | |
| Other reagents | |||
| Methoxyflurane (Pentrhox) | Medical Developments International | Not applicable | |
| Equipment and plasticware | |||
| Oxoid Anaerobic Jars | Thermo Fischer Scientific | HP0011/HP0031 | |
| COPAN Pasteur Pipettes | Interpath Services | 200CS01 | |
| Eppendorf 5810R centrifuge | Collect bacterial pellets by centrifugation at 2,200 rpm for 10 mins at 4°C | ||
| 23g precision glide needle | BD Bioscience | 301805 | |
| Parafilm M | Bemis, VWR | PM996 | |
| Portex fine bore polythene tubing | Smiths Medical | 800/100/200 | |
| Plastic feeding catheters | Instech Laboratories | FTP20-30 | |
| 1 ml tuberculin luer slip disposable syringes | BD Bioscience | 302100 | |
| Eppendorf micropestle for 1.2 – 2 mL tubes | Sigma Aldrich | Z317314 | Autoclavable polypropylene pestles used for stomach homogenization |
| Sterile plastic loop | LabServ | LBSLP7202 |
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Citazione di questo articolo
An Intragastric Gavage Technique for Controlled Helicobacter Infection in Mice. J. Vis. Exp. (Pending Publication), e21710, doi: (2023).