Helminths are one of the major health problems of working donkeys, often with heavy worm burden and contributing to their early demise and/or reduction in their work output. Cross-sectional and longitudinal studies were conducted to investigate the current infection prevalence and level of strongyles infection donkeys would acquire through different seasons in the mid-lowland agro-ecological zones of Ethiopia. For this purpose, faecal samples from 206 (cross-sectional study) and 102 (longitudinal study) randomly selected donkeys were directly collected from the rectum and analysed. For the longitudinal study, the 102 donkeys dewormed at the end of main rainy season, beginning of October, were monitored for the level of strongyle infection they would acquire during subsequent dry and short rainy seasons. The cross-sectional study of 206 donkey has revealed an overall infection prevalence of 89.3% (95% confidence interval (CI) = 84.4, 92.9). Donkeys in the lowland zone showed a significantly higher strongyle infection prevalence (P = 0.0126) and mean eggs per gramme of faces (EPG) (P = 0.001; 2775 EPG) compared to donkeys in the midland zone (980.8 EPG). Age, sex and body condition did not have any significant effect on either the infection prevalence or level of infection (P > 0.05). The longitudinal study has shown a significantly lower strongyle infection prevalence (P = 0.003) and level of infection donkeys acquired (P = 0.001) in the subsequent dry and short rainy seasons compared to the main rainy season following October deworming. However, these values were not significantly different between the two agro-ecological zones (P > 0.05). This study clearly showed that parasitic infections are primarily acquired during the main rainy season when pasture/herbage coverage is relatively better, and the environment is conducive for parasites survival and development. On the other hand, the finding of majority of donkeys shedding low or no eggs during the dry and short rainy seasons showed that October deworming was effective, and donkeys acquire low or no parasitic infection during the subsequent dry and short rainy seasons. Therefore, the practice of anthelmintic treatment of donkeys at the end of short rain in May may not be necessary, and October deworming once a year is sufficient.