Objective: Assessment of influenza virus pathogenesis through intranasal inoculation of mice with varying viral doses, monitoring body weight changes and survival
Gather these items before starting the experiment. Check off items as you prepare.
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Generated by reverse genetics • Not applicable • Not specified • Not mentioned
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CDC • Not applicable • Not specified • Not mentioned
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Measure and record baseline body weights of all mice prior to viral inoculation
Note: Essential for determining 25% weight loss threshold for euthanasia
“Baseline body weights were measured prior to infection”
Anesthetize mice with isoflurane prior to intranasal inoculation
Note: Required for safe intranasal inoculation procedure
“Three mice per group were anesthetized with isoflurane and intranasally inoculated”
Intranasally inoculate anesthetized mice with 50 µl of viral suspension at doses of 10^2, 10^3, 10^4, or 10^5 PFU of CA04 and KUTK-4, or undiluted virus stocks (CA04; 10^6.5 PFU, KUTK-4; 10^6.6 PFU)
Note: Three mice per group for survival monitoring; 6 mice per group for virologic and pathologic examinations
“Three mice per group were anesthetized with isoflurane and intranasally inoculated with 10^2, 10^3, 10^4, or 10^5 PFU (50 µl) of CA04 and KUTK-4, or undiluted virus from virus stocks”
Monitor and record body weight of all mice daily for 14 days post-infection
Note: Mice with body weight loss exceeding 25% of pre-infection values must be euthanized
“Body weight and survival were monitored daily for 14 days and mice with body weight loss of more than 25% of pre-infection values were euthanized”
Monitor and record survival status of all mice daily for 14 days post-infection
Note: Record any deaths and their timing
“Body weight and survival were monitored daily for 14 days”
Euthanize 3 mice per group on days 3 and 6 post-infection for virologic and pathologic examinations
Note: 6 mice per group total were intranasally infected with 10^5 PFU of S-OIVs and KUTK-4
“For virologic and pathologic examinations, 6 mice per group were intranasally infected with 10^5 PFU of S-OIVs and KUTK-4 and 3 mice per group were euthanized on days 3 and 6 pi”
Determine virus titres in various organs from euthanized mice using plaque assays in MDCK cells
Note: Performed on tissue samples from mice euthanized on days 3 and 6 post-infection
“The virus titres in various organs were determined by plaque assays in MDCK cells”
Baseline body weights measured prior to infection