There is growing evidence showing that spinal manipulation increases muscle strength in healthy individuals as well as in people with some musculoskeletal and neurological disorders. However, the underlying mechanism by which spinal manipulation changes muscle strength is less clear. This study aimed to assess the effects of a single spinal manipulation session on the electrophysiological and metabolic properties of the tibialis anterior (TA) muscle. Maximum voluntary contractions (MVC) of the ankle dorsiflexors, high-density electromyography (HDsEMG), intramuscular EMG, and near-infrared spectroscopy (NIRS) were recorded from the TA muscle in 25 participants with low level recurring spinal dysfunction using a randomized controlled crossover design. The following outcomes: motor unit discharge rate (MUDR), strength (force at MVC), muscle conduction velocity (CV), relative changes in oxy- and deoxyhemoglobin were assessed pre and post a spinal manipulation intervention and passive movement control. Repeated measures ANOVA was used to assess within and between-group differences. Following the spinal manipulation intervention, there was a significant increase in MVC (p = 0.02; avg 18.87 ± 28.35%) and a significant increase in CV in both the isometric steady-state (10% of MVC) contractions (p < 0.01; avg 22.11 ± 11.69%) and during the isometric ramp (10% of MVC) contractions (p < 0.01; avg 4.52 ± 4.58%) compared to the control intervention. There were no other significant findings. The observed TA strength and CV increase, without changes in MUDR, suggests that the strength changes observed following spinal manipulation are, in part, due to increased recruitment of larger, higher threshold motor units. Further research needs to investigate the longer term and potential functional effects of spinal manipulation in various patients who may benefit from improved muscle function and greater motor unit recruitment.