Chen, Y, Hill, HZ, Lange, G, and Falvo, MJ. Salivary mitochondrial DNA copy number is associated with exercise ventilatory efficiency. J Strength Cond Res 31(7): 2000-2004, 2017-Mitochondrial DNA copy number (mtDNAcn) is an index of mitochondrial content and is responsive to changes in exercise training volume. Therefore, assessment of mtDNAcn may help to optimize exercise prescription and aid in athlete monitoring. Although previous work has assessed mtDNAcn derived from skeletal muscle and blood using invasive approaches, no study has examined salivary mtDNAcn and its relationship with sport performance. Fifteen adults (32.2 ± 7.1 years) volunteered to participate in this study. Each participant provided a saliva sample for the analysis of mtDNAcn via real-time polymerase reaction. In addition, participants completed an exercise challenge test to assess oxygen consumption relative to body weight (V[Combining Dot Above]O2·kg) and ventilatory efficiency (VE/V[Combining Dot Above]CO2). Using multiple linear regression, we examined the association of V[Combining Dot Above]O2·kg and VE/V[Combining Dot Above]CO2 with salivary mtDNAcn, adjusting for self-reported physical activity (min·wk). Greater mtDNAcn was associated with lower VE/V[Combining Dot Above]CO2 (p < 0.01) and higher V[Combining Dot Above]O2·kg (p < 0.05). In our model adjusted for physical activity, greater mtDNAcn remained associated with lower VE/V[Combining Dot Above]CO2 (β = -0.186; 95% confidence interval [CI], -0.348 to -0.025; p < 0.05), but not with V[Combining Dot Above]O2·kg (β = -0.022; 95% CI, -0.113 to 0.063). Our findings suggest that salivary mtDNAcn is associated with ventilatory efficiency, which may reflect enhanced exercise efficiency as a consequence of greater total mitochondrial content. As saliva collection is noninvasive, stable at room temperature, and less costly in comparison to skeletal muscle and blood, future studies may consider using saliva for the evaluation of mitochondrial content for the purposes of monitoring exercise training as well as optimizing exercise prescription.