The objective of this study was to ascertain whether sequential sampling and isotopic analysis of bovine hooves could be used to reconstruct the dietary history of cattle. A controlled, on-farm experiment was conducted in which cattle were switched from a barley-based diet to an isotopically different diet incorporating maize, urea and seaweed (the isotopic spacing between diets was 13.6‰ for δ13C and 8.0‰ for δ15N) and maintained on that diet for 168 days. Postmortem sampling of the cleaned anterior wall of the lateral, left front claw was carried out on five individuals using a micro-drilling technique. From the first 60 mm of each claw, up to 41 samples with a spacing between them of less than 1 mm were collected. Bands were less than 1 mm deep and had a mean width of 1.2 mm. The hoof keratin showed a rapid increase followed by a slower increase in its δ13C and δ15N values following the diet switch, suggesting that C and N in hoof keratin originate from more than one pool. However, the response of the N isotope composition of the hoof was somewhat delayed compared with that of C. Estimated mean hoof growth rates for these cattle were 10.5 ± 2.3 mm per month and 6.7 ± 1.0 mm per month (±SD, n = 5) when receiving the barley-based transition diet and the maize-based experimental diet, respectively. These values are considerably higher than previous estimates obtained by visual methods and they suggest that diet may have a greater influence on hoof growth rates than seasonality. These results demonstrate that hooves are a suitable incremental tissue for high-resolution isotopic reconstruction of the dietary history of bovine animals. Copyright © 2007 John Wiley & Sons, Ltd.