Mealybugs are harmful insect pests of crop plants worldwide. It has been recognized as the primary vector of Ampelovirus spp., resulting in lower yield, quality, and productivity in pineapple production. Grey mealybugs (Dysmicoccus neobrevipes) and pink mealybugs (Dysmicoccus brevipes) have been identified as the vectors of Ampelovirus spp. in pineapples. The etiology of mealybug wilt of pineapple (MWP) disease is still unknown due to the virus's intricate interaction with mealybugs and ants. Thus, the current study sought to investigate the disease transmission mechanism of MWP. The experiment was carried out in the greenhouse between 2023 to 2024 using a fully randomized design (CRD) with ten replications for each treatment. The MD2 pineapple was infected with 20 viruliferous mealybugs (Treatment 1 (T1)), 20 viruliferous mealybugs, and 20 ants (Treatment 2 (T2)), and a control treatment (uninoculated). Data were observed and collected on lesion development, disease incidence, and disease severity based on symptom appearance and continued on a 14-day interval for six months. Results showed no significant differences between inoculated plants T1 and T2 based on the area of lesions and disease severity for both trials (P ≥ 0.05). However, inoculated plants from T2 produced the largest area of lesions and the highest disease severity, while inoculated plants T1 displayed the smallest area of lesions and the lowest disease severity. The survival and growth rate of the mealybug population increases with the presence of ants. The presence of ant colonies has been shown to alter mealybug behavior by facilitating fast plant colonization. In summary, mealybugs spread MWP disease of MD2 cultivar by transmitting Ampelovirus spp., and the disease severity increased with the presence of ants, who function as mealybug transporters.