This study was undertaken with the aim of determining reference physiological values for working Ethiopian donkeys in four districts of Oromiya regional state by taking vital signs (body temperature, pulse and respiration rate) and gut sounds from a total of 130 apparently healthy donkeys. .16 for female and male donkeys, respectively. There was no statistically significant (p>0.05) difference in nostril movement, abdominal movement and tracheal sound between daytime (morning Vs noon), age group and sex. However, strong and positive correlation (r=0.80; p<0.000) was observed among the three sites of determining respiration. There was statistically significant difference (p<0.000) in gut sounds heard at the right para-lumbar fossa (RPF), left para-lumbar fossa (LPF) and lower flank regions (LFR) during the times of day: more frequent and stronger at noon than in the morning. There was also a significant difference (p<0.004) between young and old age groups in the value of RPF and LPF. The effect of age, sex and time of day on the references values should be taken into consideration in making physical clinical examination and interpreting physiological parameters.