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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Swabbing the Vaginal Lumen to Quantify GBS Load
- Prepare one 1.5 ml micro-centrifuge tube per mouse by adding 100 µl of phosphate-buffered saline (PBS). Just prior to swabbing, pre-wet the swab in PBS.
- Restrain the mouse and insert the swab 10 mm into the vaginal lumen. Gently rotate the swab 4 times clockwise and 4 times counter clockwise, applying slight pressure to the vaginal wall.
- Transfer the swab to the 1.5 ml microcentrifuge tube with 100 µl of PBS. Prior to plating, vortex the microcentrifuge tube for ~15 sec to release the bacteria from the swab. Serially dilute each sample in PBS and plate 20 µl of dilutions 1:10 through 1:10,000 on differential medium agar plates prepared per the manufacturer's instructions. Incubate the plates at 37 °C for 24 hr. Group B Streptococcus (GBS) colonies will appear either bright pink or mauve in color. Other endogenous flora will be inhibited, or will appear as blue, white, or grey colonies.