Water mites are diverse aquatic invertebrates that provide potentially important ecosystem and economic services as bioindicators and mosquito biocontrol; however, little is known about water mite digestive physiology, including their diet in nature. Water mites, much like their spider relatives, liquefy their prey upon consumption. This results in the absence of morphologically identifiable prey in water mite mid-gut. Previous studies have reported associations in the field of water mites with presumed prey and laboratory observations of water mites feeding on specific organisms offered for ingestion; however, the present work aims to determine what water mites have ingested in nature based on molecular studies of gut contents from freshly collected organisms from the field. To elucidate water mite prey, we used next-generation sequencing to detect diverse cytochrome oxidase I DNA barcode sequences of putative prey in the guts of 54 specimens comprising two species of Lebertia and a few specimens of Arrenurus (2) and Limnesia (1). To our knowledge this is the first molecular study of the diets of water mites as they feed in nature. While the presence of chironomid DNA confirmed previous observations of midge larvae as part of the diets of Lebertia, we also found the DNA of diverse organisms in all four species of water mites, including the DNA of mosquitoes in 6 specimens of Lebertia and a large number of previously unknown prey, especially from oligochaete worms. These studies thereby reveal a greater diversity of prey and a potentially broader significance than previously appreciated for water mites in aquatic food webs. Molecular studies like this can detect water mite predators of mosquito larvae and add knowledge of water mite predatory contributions to freshwater food webs.