This study evaluated the effect of roughage:concentrate (R:C) ratio associated with a variable particle size of physically effective neutral detergent fibre (peNDF8) in the forage (Tifton-85 hay) on the performance, carcass traits and meat quality of lambs. Seventy-two 4-month-old, non-castrated Santa Ines male lambs (23.5 ± 2.32 kg BW) were distributed in a completely randomized design, in a 2 × 2 factorial arrangement two peNDF8 hay particle sizes (13 and 6 mm) and two R:C ratios (700:300 and 500:500 g/kg DM total). DMI, DM, NFC and TDN digestibility's, N-intake and N-faecal excretion were affected by the R:C ratio (P < 0.05). However, the N-retained was not affected by the studied variables (P > 0.05). It was observed an interaction (P < 0.05) between the peNDF8 and R:C ratios for final BW, average daily gain (ADG), colour parameters and pH 24 h. The lower roughage ratio provided greater (P < 0.05) concentrations of C14:1, C16:1–cis9, C18:1–cis9, ΣMUFA, Σn–6:Σn–3 and hypocholesterolemic/hypercholesterolemic index, enzymatic activity Δ9desaturase-C16 and -C18. Lambs fed a lower roughage diet had improved performance and feed efficiency, however, presented reduced polyunsaturated fatty acids (PUFA) concentrations in the meat, especially Σn–3 family. Higher roughage diet and larger peNDF8 particle size improved the concentrations of PUFA while decreased Σn–6:Σn–3 ratio in meat. Larger peNDF8 particle size associated with higher roughage proportion, have reduced animal performance however, it increased protein concentration, a* and C* colour parameter without affecting fatty acids profile of Longissimus lumborum muscle.