The purpose of this study was to determine the effects of plasmid-mediated growth hormone releasing hormone (GHRH) supplementation on the clinical outcomes of pigs vaccinated against and challenged with either Mycoplasma hyopneumonia (M. hyo) and/or with porcine reproductive and respiratory syndrome (PRRS) virus. Before the first vaccination, pigs received a single i.m. injection of 0.625 mg of a porcine GHRH-expressing plasmid followed by electroporation of the injection site. Pigs were vaccinated at 2-wk intervals, challenged with either M. hyo and/or PRRS virus 2-wk after the second vaccination, and necropsied at 17 and 36 d after challenge. Clinical parameters associated with M. hyo challenge were improved with the GHRH treatment. Average daily gain between challenge and necropsy was improved (P = 0.04). Respiratory scores for M. hyo-challenged pigs tended to be lower in GHRH-treated animals compared to controls, and coughing scores were improved by the treatment (P = 0.01). Macroscopic lesions associated with M. hyo infection pneumonia were fewer in the group that received the GHRH-expressing plasmid. No differences between treatment groups in the macroscopic pneumonia associated with PRRS virus were observed. No differences in serum antibodies to M. hyo or PRRS virus were observed with GHRH treatment. Nevertheless, IgG in the bronchioalveolar lavage was increased by the GHRH treatment in M. hyo-challenged animals (P < 0.03). The results of this study suggest that GHRH supplementation before vaccination may enhance the protection against M. hyo-induced pneumonia and that a single dose of GHRH-expressing plasmid was sufficient to elicit an improved clinical outcome in this disease challenge model.