Jun 03, 2025
Acclimatization Plan for Rats – Daily Protocol (3 Weeks)
- Yalei zang1
- 1Charité
Protocol Citation: Yalei zang 2025. Acclimatization Plan for Rats – Daily Protocol (3 Weeks). protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9oxopv3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 31, 2025
Last Modified: June 03, 2025
Protocol Integer ID: 219246
Keywords: acclimatization plan for rat, week acclimatization protocol, acclimatization plan, combining gradual habituation, gradual habituation, daily protocol, rat, environmental enrichment
Funders Acknowledgements:
Yalei Zang
Abstract
We developed a 3-week acclimatization protocol combining gradual habituation, environmental enrichment. The protocol improves data quality and animal welfare and is compatible with randomized, blinded, and transparent study designs.
Troubleshooting
Introduction
General Considerations for a 3-Week Acclimatization Period
- Purpose: Ensure animals adapt to new housing, handling, and study-related procedures.
- Regulatory Compliance: Follow FELASA, ARRIVE guidelines, and institutional animal welfare regulations.
- Environmental Familiarization: Gradual exposure to handling, noise levels, and experimental procedures to reduce stress responses.
Week 1 – Basic Adaptation & Health Monitoring
Goal: Allow rats to adapt to their new environment, minimize transport stress.
| A | B | C | |
| Day 1 | Arrival & Housing | • Group housing (7–8 per cage) with bedding, nesting material, tunnels. • Health check: fur, eyes, activity, hydration, respiration. • Introduce minimal human contact by keeping hands in the cage, no active engagement, let rats approach. • Offer peanuts as a treat in the end. • Observe social behavior, document findings. | |
| Day 2 | Behavioral Observation | •Observed social behavior, aggression, grooming, and activity levels. • Brief (10–15 sec) cupping with rats showing interest in human interaction. • Treat: One teaspoon of oats offered per cage. | |
| Day 3 | Behavioral Observation | •Performed partial bedding changes in preparation for the weekend. • Brief (10–15 sec) cupping and rat tickling with two hands. • Each rat was transferred to a new cage after tickling. • Treat: Peanuts offered at the end. | |
| Day 4 | Behavioral Observation | • Increased handling duration to ~1 minute per rat. • Minimal restraint to allow voluntary interaction. | |
| Day 5 | Behavioral Observation | ||
| Day 6 | Behavioral Observation | • Increased handling duration to 1–2 minutes per rat, including cup cuddling and tickling. • Treat: One teaspoon of oats per cage. | |
| Day 7 | Behavioral Observation | • Planned exposure to routine facility sounds (e.g., door opening, cage transport on roller cargo, equipment beeping). • Observed activity levels, weight stability, and signs of stress |
Week 2 – Handling and Refinement
Goal: Familiarize rats with handling, introduce a homemade small playground for social play in a larger group, and begin non-invasive procedure acclimatization (weight measurement, exposure to anesthesia induction box without anesthesia).
| Day 8 | Extended Handling & Social Play | Increase handling duration to 2–3 min per rat. Introduce rats to the homemade small playground (allowing larger group interaction outside cages). Observe social interactions. | |
| Day 9 | Weight Measurement Introduction | Place each rat briefly on a digital scale, allowing voluntary exploration. Reward calm behavior with treats. | |
| Day 10 | Acclimatization to Anesthesia Induction Box | Introduce the anesthesia induction box without administering anesthesia. Allow exploration inside to reduce stress for future procedures. | |
| Day 11 | Handling & Second Playground Session | Repeat extended handling (2–3 min per rat), followed by another social play session in the larger playground. Observe interactions. | |
| Day 12 | Rest Day | No handling, only routine health monitoring. | |
| Day 13 | Handling Refinement & Enrichment | Increase handling time if tolerated, continue interaction training. Introduce a new enrichment object. | |
| Day 14 | Weight Training & Repeat Box Acclimation | Repeat weight measurement, ensuring rats voluntarily enter the scale. Second exposure to the anesthesia induction box. |
Week 3 – Experimental Procedure Acclimatization
Goal: Familiarize rats with noise exposure to the echocardiography room, metabolic cage acclimation, and final health monitoring (blood sample collection on the last day).
| Day 15 | Noise Acclimatization – Echocardiography Room | Expose rats to the echocardiography room sounds (machine beeping, probe setup, researcher presence) for 10–15 min. Observe stress responses. | |
| Day 16 | Metabolic Cage Acclimation (Short Exposure) | Place rats in metabolic cages for 30 min. Observe behaviors, assess stress levels. | |
| Day 17 | Second Noise Acclimatization & Handling | Repeat echocardiography room exposure, increasing duration. Continue handling. | |
| Day 18 | Extended Metabolic Cage Exposure | Increase time in metabolic cages to 1 hour. Monitor food/water intake and activity. | |
| Day 19 | Rest Day | No handling, only routine health checks. | |
| Day 20 | Final Echocardiography Room Visit | Ensure all rats tolerate the environment well. Observe behavior. | |
| Day 21 | Final Monitoring & Blood Sample Collection | Weigh all rats and collect blood samples for baseline assessment before the study begins. |
Protocol references
Percie du Sert N, Ahluwalia A, Alam S, Avey MT, Baker M, et al. (2020) Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0. PLOS Biology 18(7): e3000411.