Mouse inoculation test (MIT) is a technique widely used for rabies diagnosis and must be liable to refinement
due to animal welfare. The present study aims to compare five different anesthetic associations to stablish a
protocol to improve the MIT procedure suitable for animal welfare and safe for a routine of viral isolation in
newly weaned mice (3 weeks of age). 80 Swiss-Webster mice (Mus musculus) - 40 females and 40 males, 3-weekold, weight ranging from 11 to 14 grams – were used to conduct all procedures. Five anesthetic associations
were tested: KX (Ketamine 100 mg/kg and Xylazine 10 mg/kg), KXA (Ketamine 80 mg/kg, Xylazine 5 mg/kg, and
Acepromazine 1 mg/kg), KXT (Ketamine 80 mg/kg, Xylazine 5 mg/kg, and Tramadol 5 mg/kg), KXAT (Ketamine
100 mg/kg, Xylazine 10 mg/kg, Acepromazine 2 mg/kg and Tramadol 5 mg/kg) and ATI (Acepromazine 1 mg/kg
+ Tramadol 5 mg/kg + Isoflurane 5% - 0.5 L/min for induction and 2.5% - 0.5L/min for maintenance). Injectable
anesthesia was administered intraperitoneally. We monitored the respiratory rate and body temperature.
Response to anesthesia was evaluated according to the induction, surgical anesthesia, and recovery periods.
The KXAT and ATI protocols induced surgical anesthesia, with the ATI protocol being the most appropriate and
safe to perform the MIT procedure with 100% efficiency, absence of mortality, and rapid recovery of respiratory
rate and temperature in the period after the procedure.