Standard Operating Procedure for Operation of Mouse Breeding

1. Purpose

This SOP describes standardized procedures for breeding and maintaining laboratory mice, including mating schemes, pregnancy management, nutritional support, pup care, weaning, and transfer for experimental use.

2. Scope

This SOP applies to all staff in the animal breeding facility responsible for mouse husbandry and breeding operations.

3. Responsibilities

• Animal Facility In-Charge: Supervise breeding program and ensure compliance with guidelines.
• Animal Technicians: Perform daily husbandry, mating setups, monitoring, and records.
• Veterinarian: Confirm pregnancy, monitor health, and approve transfer to laboratory.

4. Materials and Equipment

• Individually ventilated cages (IVCs) or polycarbonate cages with bedding (autoclaved corn cob/wood shavings).
• Water bottles with sipper tubes.
• Standard laboratory mouse pellet diet.
• Nesting material (tissue paper, cotton pads).
• PPE (gloves, mask, gown).
• Cage cards and breeding records.
• Disinfectants for cleaning.

5. Procedure

5.1 Housing and Mating Schemes

1. Maintain environmental conditions: Temperature 20–24 °C, Humidity 40–70%, Light cycle 12:12 h (light:dark).
2. Mating setups:
• Monogamous mating (1 Male : 1 Female) – ensures accurate pedigree.
• Polygamous mating (1 Male : 2 Females) – increases efficiency.
• Harem mating (1 Male : 3 Females) – used for large-scale breeding; requires close monitoring.
3. After successful mating (evidence of copulation plug or pregnancy signs), remove male within 5–7 days to avoid stress or cannibalism risk.

5.2 Male Mouse Separation

• Transfer males to male-only housing cages after mating period.
• Clearly label cages with Male ID, separation date, and last mating record.
• Never keep adult males together if aggressive—house singly or in compatible groups.

5.3 Pregnancy Confirmation

• Copulatory plug: White plug in vaginal opening observed within 24 hours of mating (early indicator).
• Body weight gain: Gradual weight increase visible by day 10–12.
• Palpation: Possible around day 12–14.
• Gestation period: 19–21 days (average 20 days).

5.4 Nutrition and External Supplements

• Provide standard mouse pellet diet ad libitum.
• Provide clean drinking water daily.
• During pregnancy and lactation:
• Add protein and energy supplements (sunflower seeds, soaked gram, boiled egg pieces, or commercial high-protein pellets).
• Ensure nesting material is available to reduce maternal stress.

5.5 Parturition and Pup Care

• Average litter size: 6–12 pups.
• Do not disturb dams during delivery.
• Avoid cage cleaning for first 5 days after parturition unless absolutely necessary.
• Observe quietly for maternal care and pup health.

5.6 Weaning and Pup Separation

• Weaning period: 19–21 days post-birth.
• Procedure:
• Gently separate pups from mother and house in clean cages.
• Segregate by sex (male and female separately) to prevent premature breeding.
• Provide pellet diet and water.
• Observe pups for 2–3 days post-weaning for adaptation.

5.7 Transfer of Mice to Laboratory

• Weaned mice can be transferred to the laboratory only after veterinary approval.
• Cages must be labeled with Animal ID, strain, sex, date of birth, and transfer date.
• Handle mice gently to minimize stress during transfer.

5.8 Mouse Maturity for Experimental Use

• Sexual maturity:
• Females: 6–8 weeks.
• Males: 8–10 weeks.
• Mice should be healthy and fully mature before being used in experiments.

6. Record Keeping

Maintain detailed records for:

• Mating dates and mating scheme used.
• Pregnancy confirmation method and date.
• Litter size at birth.
• Weaning date and number of pups.
• Transfer to laboratory.

7. Health and Hygiene

• Maintain strict cleanliness and cage sanitation twice weekly.
• Monitor all animals daily for signs of stress, fighting, or illness.
• Report any abnormalities to the veterinarian.

8. Safety and Ethics

• Follow CCSEA/IAEC guidelines (or relevant national guidelines).
• Handle mice with care using correct restraint techniques.
• Ensure ethical use and proper welfare standards.

This SOP ensures efficient mouse breeding, healthy litters, and reliable supply of experimental animals.

                                         END OF THE DOCUMENT

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