Section -II
Protocol form for research proposals to be submitted to the Institutional
Animal Ethics Committee/ CPCSEA, for new experiments or extensions of
ongoing experiments using animals.
1. Project / Dissertation / Thesis Title:
A comparative study of the effect of Moringa oleifera (MO) leaf extract and
its nanoformulation on oxidative stress markers in AlCl₃-induced
Alzheimer’s disease (AD) in Wistar rats.
6. Describe details of study plan to justify the use of animals
Objective:
1. Induction of Alzheimer's in Wistar Albino rats by using AlCl₃
2. Assessment of behavioural changes and biochemical tests
3. To evaluate and compare the effects of crude extract of MO and
nanoformulation by studying the histopathology of the brain in control and
treatment groups.
Study plan: After acclimatisation of 1 week, 36 animals are divided into 6
groups, i.e., normal control group, disease control group, placebo group, MO
extract treatment group, low-dose MO nanoformulation group, and the same dose
of MO nanoformulation group as that of crude extract (n = 6). Except for the
normal control group, other group animals will be administered AlCl₃ (100
mg/kg b.w.) orally daily for 28 days, and treatment groups will receive their
respective treatment doses, i.e., the MO extract treatment group (250 mg/kg
b.w.), the MO nanoformulation low-dose group (150 mg/kg b.w.), and the MO
nanoformulation group (250 mg/kg b.w.) for 28 days. After the 28th day, a
cognitive test is being conducted, and the oxidative stress markers of the brain
homogenate will be evaluated.
7. Animals required:
a. Species and Strain : Wistar albino rats
b. Age and Weight : 180 to 220 gm
c. Gender : Both Male and Female
d. Number to be used : 36
Animals required for Pharmacological activity
Year Group No. of animals required
2025 Normal control (NC) 6
Disease control (DC) 6
Disease control + Placebo 6
Disease control + MO extract 6
Disease control + low dose 6
nanoformulation of MO
Disease control + same dose 6
nanoformulation of MO
Total no. of animals required 36
e. Number of days each animal will be housed: 35 days
9. Describe the procedures in detail:
a. Describe all invasive and potentially stressful non-invasive procedures that
animals will be subjected to in the course of the experiments
The drugs will be administered to the animal through the oral route by
using the feeding tube, and at the end of the study, the animals will be
euthanised as per the CPCSEA guidelines for tissue collection.
b. Furnish details of injections schedule Substances: Not applicable
Doses :
Sites :
Volumes :
c. Blood withdrawal Details: Not applicable
Volumes :
Sites :
d. Radiation (dosage and schedules): Not applicable
e. Nature of compound/Broad Classification of drug/NCE: Not applicable
12. Describe post-experimentation procedures.
a. Scope for Reuse: After completion of the experiment, all the animals will
be euthanised for tissue collection, so there is no question of reuse of animals.
b. Rehabilitation (Name and Address, where the animals are proposed to be
rehabilitated): Not applicable
c. Describe method of Euthanasia (If required in the protocol): All the
animals will be euthanised by administering a 100 mg/kg dose of ketamine.
d. Method of carcass disposal after euthanasia: Incineration
