The demand for a reduced use of pesticides in agriculture requires the development of specific strategies for managing arthropod pests. Among eco-friendly pest control tools, pheromone-based mating disruption (MD) is promising for controlling several key insect pests of economic importance, including many lepidopteran species. In our study, we evaluated an MD approach for managing the honeydew moth (HM), Cryptoblabes gnidiella, an emerging threat for the grapevine in the Mediterranean basin. The trials were carried out in two study sites, located in Tuscany (central Italy, years 2017–2019) and Apulia (southern Italy, years 2016 and 2018–2019), and by applying MD dispensers only in April, in April and July, and only in July. To evaluate the effects of MD, infested bunches (%), damaged area (%) per bunch, and number of living larvae per bunch were compared among plots covered with MD dispensers, insecticide-treated plots (Apulia only), and untreated control plots. Male flights were monitored using pheromone-baited sticky traps. Except for the sampling carried out in Tuscany in 2018, where HM infestation level was very low, a significant difference was recorded between MD and control plots, both in terms of HM damage caused to ripening grapes and/or number of living larvae per bunch. Overall, our study highlighted that MD, irrespective of the application timing, significantly reduced HM damage; the levels of control achieved here were similar to those obtained with the application of insecticides (no MD). However, MD used as stand-alone strategy was not able to provide complete pest control, which may instead be pursued by growers with an IPM approach.