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.

Declarações

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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An Intragastric Gavage Technique for Controlled Helicobacter Infection in Mice. J. Vis. Exp. (Pending Publication), e21710, doi: (2023).

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