Busseola fusca and Fusarium verticillioides are considered the most important pest and pathogen, respectively, of maize in South Africa. Busseola fusca tunnels into stems and ears of maize plants, whereas F. verticillioides causes Fusarium ear rot and deposits fumonisins in maize kernels. Frass (excreta) deposited by B. fusca larvae in maize stems and ears during feeding could potentially be contaminated with pathogens such as F. verticillioides, thereby indirectly and passively contributing to plant diseases. The mycoflora present in frass of B. fusca larvae was thus investigated in this study, and the dissemination of fumonisin-producing Fusarium spp. by B. fusca larvae in maize stems determined. Busseola fusca frass was collected from plants with visible insect damage in three maize-growing districts in South Africa. The mycoflora in the frass was isolated and identified morphologically to genus level, followed by the sequencing of target genes for species identification. Species of Acremonium, Aspergillus, Fusarium, Mucor, Rhizopus and Talaromyces were associated with the B. fusca frass. The role of B. fusca larvae in disseminating fumonisin-producing Fusarium spp. was then determined in greenhouse and field trials. Maize whorls were inoculated with F. verticillioides MRC826 spores 4 weeks after plant emergence, and infested with B. fusca larvae 2 days later. The stems were split open after 3 weeks and frass collected from feeding channels, thereafter, target DNA of fumonisin-producing Fusarium spp. was quantified using real-time PCR. Target DNA of fumonisin-producing Fusarium spp. was significantly higher in frass collected from greenhouse plants inoculated with F. verticillioides than in frass collected from the uninoculated control, indicating that the inoculation was successful in the absence of soil-borne inoculum. Nevertheless, the field trial showed no significant differences in target DNA in frass from inoculated and non-inoculated plants. This is possibly due to natural F. verticillioides infection of maize plants in the field. This study further indicated that B. fusca frass also contained maize pathogens such as Aspergillus spp. and Fusarium spp., which could cause ear rot diseases. The occurrence of Acremonium zeae in frass has potential implications for the biological control of F. verticillioides, as the fungus produces pyrrocidines A and B antibiotics which are known to be antagonistic to F. verticillioides. The occurrence of Aspergillus niger in frass requires further investigation in order to clarify its role in fumonisin contamination of maize in South Africa.