Johne’s disease (JD) is a fatal chronic enteritis which causes detrimental effects on production and health and significantly reduces animal welfare. Control of JD is highly desirable, but single milk ELISA testing may not be sensitive enough to identify all affected animals, particularly in the early stages of the disease. The objective of this study was to compare the activity of cows with a low risk of JD to cows with a high risk of JD around calving and peak lactation. The study was conducted at Harper Adams University, UK, using 42 multiparous (3.1 ± 0.22 (Mean ± SEM); range: 2–7 lactations) Holstein Friesian cows, fitted with an IceQube accelerometer (IceRobotics Ltd., Edinburgh, UK) on the back left leg. The sensors recorded lying duration (h/d), frequency of lying bouts (LB/d), average lying bout duration (LBD; min/bout) and steps (S/d). Four times during 2015 and 3 times during 2016 the cows were milk sampled, and subsequently tested for JD using an ELISA. Cows in infection groups JD0-JD2 were classed as low risk and cows in infection groups JD3-JD5 were classed as high risk. Low risk cows (JD0; n = 21 (repeat ELISA –iv)) were matched to high risk cows (JD5; n = 19 (repeat ELISA +ve), JD4; n = 2 (last ELISA +ve)) based on parity and age and there was no difference in milk yield between low and high risk JD cows (P > 0.05; 41.8 ± 1.11 vs. 43.1 ± 1.13 kg/d, respectively). Activity data were averaged and analyzed 14d before calving and peak lactation (8 wks post calving). One way ANOVA (Genstat, 18th edition, VSN International Ltd., UK) revealed no significant differences (P > 0.05) in activity between low vs. high risk cows around calving. However, at peak lactation low risk JD cows spent, on average 93 min/d longer lying down than high risk cows (P = 0.019; 11:05 vs. 09:32 hh:mm/d, respectively) and they had more LB/d (P = 0.027; 12.7 vs. 10.2 LB/d, respectively). There was no difference in average LBD (P = 0.392; 00:55 vs. 00:59 hh:mm/bout) or total S/d (P = 0.388; 1586.4 vs. 1796.9 S/d, respectively) around peak lactation. The results show that activity data from leg-mounted accelerometers have the potential to help identify cows at a higher risk of JD although more research is required.