To determine whether a single acute preexercise bout of partial-body cryotherapy (PBC) enhanced maximal-effort shuttle run performance, salivary enzyme concentration, and self-reported performance readiness. A total of 18 male rugby league players (age = 20.1 [0.5]y; mass = 91.4 [12.4]kg) were exposed to either PBC for 3minutes at -136°C (1°C) or a control condition prior to a continuous, high-intensity 6 × 40-m shuttle run test. Passive saliva samples were collected to determine salivary alpha amylase (sAA) concentration. Perceived performance readiness and well-being questionnaires were completed using a 1-to-7 Likert scale. The PBC exposure did not elicit a greater improvement in 6 × 40-m shuttle run performance in comparison with the control condition (standardized difference; +0.4 [5.9%]; P = .881; mean ± 90% confidence limits). The increase in sAA concentration was moderately greater 15minutes after PBC compared with the control group (+67 [32%], P = .012) and remained moderately higher up to 2hours post-PBC exposure compared with the control condition (+41 [40%], P = .045). There were greater improvements in self-reported perceptions of muscle soreness (+0.6 [0.4%], P = .043; units ±90% confidence limits) and mood (+0.6 [0.7%], P = .038) after PBC compared with control. It appears that a single 3-minute bout of PBC does not augment maximal effort shuttle run performance in elite rugby league players. Beneficial increases in sAA concentration, coupled with improved perceptions of muscle soreness and mood, should be explored further for alternative training or precompetition practices.