Estimation of indigestible neutral detergent fibre (iNDF) is necessary for accurate and precise predictions of feed energy values and potential microbial protein from digested NDF in the rumen. Due to lengthy laboratory procedures, iNDF has been estimated using the formula ADLÃƒ?2.4 (iNDF2.4). The relationship between iNDF and acid detergent lignin (ADL) is more variable, across and within forage species. The purpose of our study was then to assess the variability of iNDF and respective implications on ration fine-tuning for dairy cattle. Sixty forages, including grasses, maize silages and lucerne hays, were fermented in vitro from 0 to 240 hours. Residual NDF of the fermented samples were obtained at 0, 6, 12, 24, 36, 48, 72, 96, 120, 144, 216 and 240 h, with the last value assumed to represent iNDF (iNDF240).This was used to obtain the potentially digestible NDF fraction (pdNDF). Rates of digestion of pdNDF were obtained assuming a first order decay. Simulations with the Cornell Net Carbohydrate and Protein System (CNCPS v 6.1, 2012) were done to evaluate the effects of the different estimated iNDF and NDF rate of digestion (kd) on energy and microbial protein estimations, assuming the requirements of a high-yielding lactating cow and a standard TMR with at least 50% forage. Results were dependent on the amount of forage and respective NDF and ADL. The iNDF240 values resulted between 1% and 136% higher than the iNDF2.4 values. The reduced pdNDF pool resulted in both lower cell wall linked protein in the rumen and microbial protein of around 5 to 165 g, and, as a consequence, on a total decreased metabolizable protein for milk. Use of iNDF240 showed consistently lower metabolizable energy (ME) between 2 and 10 MJ/day, compared to when using iNDF2.4. The improved metabolizable protein (MP) and ME values would result in 0.3 to 3.2 kg/d less milk when using iNDF2.4. This research demonstrates how points later in the fermentation curve, even if not biologically relevant for the cow, result in a more accurate and precise estimation of the rate of NDF digestibility. Indigestible NDF estimated at 240 h would give better predictions of rumen parameters in models like the CNCPS and better fine-tuning in dairy cow diets, especially when using high forage and/or NDF rations.