The objectives of this preliminary investigation were to evaluate the feasibility of transrectal colour Doppler sonography (TCDS) for determining blood flow of the pudendoepigastric trunk in cows with experimentally induced Escherichia coli Mastitis. Five primiparous Holstein dairy cows, 4–6 months after calving, were examined in two trials. All monitored udder quarters were initially clinically healthy, somatic cell count (SCC) was <50 000 cells/ml and bacteriologically negative. The blood flow of the left and the right pudendoepigastric trunk was described by the blood flow volume (BFV). In the methodological part of the study, the intra-observer precision of the method was evaluated. The coefficients of variation of the BFV were 7·1% for the left and 9·4% for the right pudendoepigastric trunk. The intraclass correlation coefficients of the BFV were 0·99 (P<0·001) for the left and 0·75 (P=0·004) for the right vessel. BFV did not differ significantly between the left and the right side nor between pre- and post-milking nor between oestrus and dioestrus. In the experimental part of the study, significant differences of increasing BFV between 0 and 12 h p.i. (post infectionem) (P=0·043) and decreasing BFV between 12 and 24 h p.i. (P=0·043) were discovered for the pudendoepigastric trunk of the infected right side. In the left-right (control-infection) comparison a significant increase of the right BFV was observed at 12 h p.i. (P=0·043). The difference of an increasing SCC correlated positively with the difference of an increasing BFV between 0 and 12 h p.i. (Spearman's ρ=1·00; P=0·043) for the right infected side. It was shown that TCDS is a reproducible technique for investigating pathological mammary blood flow changes at an early stage of acute mastitis.