This study investigated the effect of restricting nasal breathing during a series of 20-m shuttle runs. Ten male participants (mean age = 21.7 ± 2.4 years, height = 1.80 ± 0.62 m, mass = 79.2 ± 10.4 kg, sum of 4 skinfolds = 54.5 ± 7.8 mm) were required to either (a) dive on the ground and complete a rolling sequence (condition = GRD) or (b) complete the shuttles while staying on their feet and tagging the line with 1 foot, at the end of each 20-m segment (condition = STD). The shuttle runs were completed with and without a nose clip (no clip = nc; with a clip = clip) under 4 different trial conditions in a randomized order (GRDnc; GRDclip; STDnc; and STDclip), requiring the participants to return on 4 separate occasions separated by 5-7 days. Heart rate was recorded throughout each trial, and the rate of perceived exertion (RPE) was measured at the completion of each shuttle sequence. Pretrial and posttrial lactate and respiratory function measures were also recorded. The general linear model with repeated measures analysis indicated that there was a significant effect for Roll (GRD > STD) (p ≤ 0.05) but not for Clip (p > 0.05) on total time to completion in the trials. There was no significant interaction of the conditions (Roll × Clip) for RPE (p > 0.05). Similarly, there was no significant effect for blood lactate measured 3 minutes post the last shuttle for Roll (p > 0.05) and Clip (p > 0.05). There was a significant main effect on the HR across all 6 time points (i.e., pre, intervals 1-4 and 10 minutes post) (p ≤ 0.05) and for Roll (GRD > STD) (p ≤ 0.05), but not for Clip (p > 0.05). No significant effect of Roll or Clip was found for any of the recorded ventilation measures (p > 0.05). On the basis of these findings, the use of restricted nasal breathing, while performing a high-intensity shuttle sequence as a method of increasing the acute training effect on athletes, is questionable, so strength and conditioning coaches should carefully consider their rationale for using such a training strategy.