Step Procedure accomplished Influenza A virus PR8 (H1N1) Equipment Data Collected 1 H1N1 virus cultured and infectivity titrated - incubator, centrifuge,cell counter - 2 Mice weighed and anesthetized
balance scale baseline body weight 3 Mice infected intranasally 2 x 103 FFU nebulizer (1 mL syringe) - 4 Weight loss and survival monitored for 14 days - balance scale daily body weights, survival
- Balance scale
- Syringes, 1 mL
- Influenza A/Puerto Rico/8/34 (H1N1; PR8) adapted for mouse; virus propagated in the chorioallantoic cavity of 10d embryonated hen eggs for 48h at 37°C, and titrated according to Srivastava et al., 2009.
- Ketamine-Xylazine (Rompun®) combination anesthetic
Procedure: Influenza A virus PR8 (H1N1) infection and host response
a. Mice are weighed prior to infection (baseline).
b. Mice are anesthetized by intra-peritoneal injection of Ketamine-Xylazine combination at a dose based on individual body weight.
c. H1N1 virus is administered intranasally (2 x 103 FFU in 20 µL sterile PBS).
d. Health is monitored over a period of 14 days by assessing degree of weight loss, general health, and survival of infected mice.
e. Dead mice are noted; moribund mice or mice that have lost >25% of their body weight are euthanized and recorded as dead.
f. Strains are designated susceptible (=0) or resistant (=1), with susceptible characterized by rapid weight loss (>25%) and death within seven days of infection (Note: designations for susceptible (=0) or resistant (=1) for each strain are assigned to individual mice in the MPD dataset).
Abbreviations (see also MPD units and abbreviations) Influenza A virus PR8 FFU Focus forming units; a measure of virus titer in an infected lung PBS Phosphate buffered saline
Data collected by investigator
• percent change in body weight
• susceptibility to viral infection