Step Procedure Influenza A virus PR8 (H1N1) Equipment Data Collected 1 H1N1 virus cultured and infectivity titrated - incubator - 2 Mice weighed and anesthetized
balance scale baseline body weight 3 Mice infected intranasally 2 x 103 FFU nebulizer (1 mL syringe) - 4 Body weight and survival monitored for 13 days - balance scale daily body weights, survival
- Balance scale
- Syringes, needles
- Influenza A/Puerto Rico/8/34 (H1N1; PR8M) adapted for mouse; virus propagated in 10d embryonated chicken eggs.
- Ketamine-Xylazine in sterile NaCl (50 mg/mL Ketamine (Invesa Arzneimittel GmbH, Freiburg Germany); 2% Xylazine (Bayer Health-Care, Leverkusen)
- Phosphate buffered saline (PBS)
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 13 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. (It should be noted that for mice that did not show any signs of body weight loss over the entire time period after infection, additional parameters were not obtained to verify that they were infected.)
f. Strains are designated susceptible (=0) or resistant (=1), with susceptible characterized by rapid weight loss (>25%) and death (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
Data collected by investigator
- percent change in body weight
- survival (time to death in days)
- susceptibility to viral infection