Herbivorous arthropods cause immense damage in crop production annually. Consumption of these pests by insectivorous animals is claimed to be crucial to counteract their adverse effects. Bats are considered amongst the most voracious predators of arthropods, of which some species are known to consume crop pests. In vineyard dominated Mediterranean agroecosystems, several crops are damaged by the attack of insect pests. In this study we therefore aimed 1) to explore the diet and pest consumption of the lesser horseshoe bat Rhinolophus hipposideros and 2) analyse whether the composition of pest species in its diet changes throughout the season. For that, we employed a dual-primer DNA metabarcoding analysis of DNA extracted from faeces collected in three bat colonies of a wine region in Southwestern Europe during the whole active period of most pest species. Overall, 395 arthropod prey species belonging to 11 orders were detected; lepidopterans and dipterans were the most diverse orders in terms of species. Altogether, 55 pest species were identified, among them 25 are major pests and 8 are regarded as pests affecting grapevines. The composition of pest species in faeces changed significantly with season. As a whole, the results suggest that R. hipposideros acts as a suppressor of a wide array of agricultural pests in Mediterranean agroecosystems. Therefore, appropriate management measures to favour the growth of this bat populations should be considered.