This study investigated the effect of a single aqua-acupuncture (aqua-AP) treatment on wound healing after surgical ventral incision in rainbow trout, Oncorhynchus mykiss. Three experiments were conducted using 20 fish per experiment (10 aqua-AP, 10 non-acupuncture): 1) ventral incision closed with two sutures and insertion of an acoustic transmitter, 2) ventral incision closed with two sutures (no transmitter), and 3) unclosed ventral incision. Aqua-acupuncture was performed by injecting a 0.25 mL bleb of vitamin B12 - saline mixture intramuscularly at 4 points around the surgical site. Statistical differences between aqua-AP and non-acupuncture for survival, transmitter retention, wound closure and redness (inflammation) were conducted (p<0.05). Experiment 1 had significantly lower (p=0.007) redness scores with aqua-AP treatment when compared to non-acupuncture. In the second and third experiments, there were no significant differences in wound closure times or redness, and acoustic transmitter retention was not significantly different between treatments. Experiment 3 had granuloma formation (50% both groups) in place of skin edge closure at 6 weeks. By study termination (12 weeks), only 1 aqua-AP fish still had a granuloma compared to 4 fish in non-acupuncture (p<0.04). There were no significant differences in wound closure times (aqua-AP versus non-acupuncture). While the overall results of this study were inconclusive for wound healing time, the decreased redness (Experiment 1) and improved granuloma resolution (Experiment 3) suggest that there may be potential benefit from aqua-AP use in rainbow trout wound healing. Aqua-AP appears safe to use on fish with future research directed at technique refinement.