1- One investigator is listed. Will one person have the expertise to do all this work? Additionally, the supervisor is listed later as havng a role in animal care and follow up. Hence at least the supervisor(s) must be listed as investigators.

2- Who willmprepare the candidate vaccines? 

3- Vaccines will be prepared using cell lines, so the proptocol for cell lines must be completed.

4- Based on which papers and what asumprions was the sample size calculated?

5- Sample size mentions 52 mice, while later in the animal protocol it is 56. Additionally, 14x4 is 56 not 52. Please put the correct number.

6- Please justify why the investigator did not determine any possible reduction of mice numbers.

7- Is there a scientific justification for not not doing a pilot study?

8- How the risks will be metigated was not mentioned.

9- Please put a reference for the statement that female mice a best for this kind of study.