An Intragastric Gavage Technique for Controlled Helicobacter Infection in Mice

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

1. 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 selective supplement" (consisting of vancomycin, 10 μg/mL; polymyxin B, 25 ng/mL; trimethoprim, 5 µg/mL; amphotericin B, 2.5 μg/mL); and 5–10% (v/v) horse blood. T....

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 m.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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