Sarda (n = 8), Awassi (n = 8), and Merino (n = 8) ewes were subjected unilaterally to once-daily milking (ODM) or twice daily milking (TDM) to test the hypothesis that the two breeds highly selected for milk production (Sarda and Awassi) would not respond as much to a change in the frequency of milking as the Merino, a wool sheep that has not been selected for dairy production. Milk composition and somatic cell count (SCC) Were also assessed to determine if the changes in milking frequency affected milk quality. Milk yield was 24% and 18% lower in ODM udder halves than TDM udder halves in Sard a and Awassi breeds, respectively. The yield loss due to ODM was similar to that observed in Merino ewes (23%) and did not support our hypothesis. Fat content did not differ significantly in any breeds between ODM and TDM udder halves. Protein content was higher in the milk of ODM than TDM udder halves in Sarda and Merino ewes. The SCC was influenced by milking treatment only in the Saida ewes, with high values observed in the milk of ODM udder halves. The same trend was observed in the Awassi and Merino breeds, but the differences were not significant. The effects on milk yield, composition, and SCC caused by ODM were completely reversed when TDM was resumed. This suggests that in sheep flocks the milk yield losses due to short-time suppression of one daily milking, for example, on festive days, are low and temporary.