The aim of this work was to study how machine milking (MM) carried out in suitable conditions affects teat wall thickness and teat canal length and their return after milking to pre-milking conditions, in comparison with other milk extraction methods considered biological referents: lamb suckling (LS), milk removal by catheter (RC) and hand milking (HM). Three Latin square experiments were designed, using 11 ewes in the first experiment (LS), 13 in the second (RC) and 12 in the third (HM). Each of the Latin squares was divided into two periods: in the first, the left gland of each animal was machine milked and the corresponding treatment (LS, RC and HM) was applied to the right gland. Subsequently, in the second period the extraction methods were interchanged. During the experimental period, 4 sampling days were carried out (2 in each experimental period), where ultrasound scans were taken before (B) and immediately after milking (A) and at 1 (1 h), 2 (2 h), 3 (3 h), 4 (4 h), 6 (6 h), 8 (8 h) and 10 (10 h) hours after milking finished. Teat wall thickness (TWT), teat wall area (TWA), teat end area (TEWA) and teat canal length (TCL) were measured in all the ultrasound images. MM increased TWT after milking compared with RC. TWT, TWA, TEWA and TCL were lower (P<0·05) in HM than in MM. No significant differences (P>0·05) were found between LS and MM for any variable. The extraction method affected the recovery time of the variables, with total teat recovery at 6 h after RC and 4 h after HM. In the case of LS, the TEWA and TCL values were recovered sooner, as of 3 h. Teat recovery time after MM was similar to the extraction method with which it was compared in each experiment. Thus, considering the similar increase in wall thickness and their recovery time compared with the reference methods, it was concluded that machine milking, carried out in optimum conditions and respecting the time interval between milkings usually applied on sheep farms (8–12 h), would not affect teat integrity. Moreover, given the variability observed in teat thickness recovery time between the different experiments, further research should be carried out to study which factors intrinsic and extrinsic to the animal may affect the teat wall thickness and recovery time after machine milking.