The use of cactus cladodes in animal feed is well-established in semi-arid areas. The cactus Nopalea cochenillifera (L.) Salm-Dyck cladodes (Nopalea) have high acceptability amongst dairy cows and are resistant to carmine cochineal insects (Dactylopius opuntiae Cockerell), a problem in semi-arid regions, but in regions of prolonged drought, it has lower productivity compared with the cactus Opuntia stricta (Haw.) Haw cladodes (Opuntia), which is also resistant to the insect. The objective of the current study was to evaluate the intake and content of digestible material of dry matter (DM) and its components, feeding behaviour, microbial protein synthesis, nitrogen balance, blood parameters, performance and milk composition of Holstein cows fed a control diet, containing either Nopalea or Opuntia associated with different concentrate levels (225, 275, 325 and 375 g/kg). Ten cows with an initial average milk production of 20 ± 2.1 kg/day were distributed into a double 5 × 5 Latin square design. Diets containing 775 g roughage/kg and 225 g concentrate/kg promoted similar responses to the analysed variables regardless of the cactus cladode used, except for digestibility of neutral detergent fibre. Diets containing higher proportions of concentrate (325 and 375 g/kg) promoted greater DM intake and 3.5% fat-corrected milk yield. The diet containing Opuntia at 775:225 g/kg roughage:concentrate proportion is as effective as the control diet for Holstein cows producing 20 kg of milk/day. To promote greater milk production, higher proportions of concentrate should be added to diets using Opuntia.