Please, add “Wistar Albino rats” to the title to be specific, instead of using a non-specific word like “in vivo”

The PI should specify the estimated funding amount “even if the study is self-funded” as this study is costly and includes a lot of work and lab analysis e.g.  

-       purchase of 45 rats (with their accommodation, feeding, care, …) ,

-       rotenone and levo-dopa

-       7 biomarkers (Reduced glutathione GSH, malondialdehyde MDA, serotonin, nor-adrenaline, dopamine, TNF-α, and IL-10) and

-       histopathology (10% paraformaldehyde and embedded in paraffin wax blocks,  staining with haematoxylin & eosin (H&E))

The PI should also confirm that he/she will be able to continue the work with self-fund.

Sample size and groups: It is not meant for blood sample volume but are needed number of rats and groups of rats which you have already written them clearly in other sections like Procedure section.

Please, correct the Gantt chart to present the time-table (schedule) of the tasks of the experiment and also the flow chart to be a representative graph of the groups of rats and flow of work in the experiment. Please, upload the corrected charts.

Does this protocol involve the use of substances that may pose any health risk (…..) to humans?
Consider toxicity of rotenone (pesticide) using MSDS.
 
No expected side effects LD50 study will be done to choose the safe dose for rats.:
There might be expected side effects will be observed in the acute toxicity study due to plant extract
 
Describe the anticipated pain or distress for animals? No pain
Parkinson’ disease which is a neurodegenerative disease causes stress
 
What will be the method of disposal of dead animals? Incineration: Please, refer to the guidelines of Occupational Safety and Health Committee at NRC.