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.

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Chironomids are one of the most biodiverse and abundant members of freshwater ecosystems. They are a food source for many organisms, including fish and water mites. The accurate identification of chironomids is essential for many applications in ecological research, including determining which chironomid species are present in the diets of diverse predators. Larval and adult chironomids from diverse habitats, including lakes, rivers, inland gardens, coastal vegetation, and nearshore habitats of the Great Lakes, were collected from 2012 to 2019. After morphological identification of chironomids, DNA was extracted and cytochrome oxidase I (COI) barcodes were PCR amplified and sequenced. Here we describe an analysis of biodiverse adult and larval chironomids in the Great Lakes region of North America based on new collections to improve chironomid identification by curating a chironomid DNA barcode database, thereby expanding the diversity and taxonomic specificity of DNA reference libraries for the Chironomidae family. In addition to reporting many novel chironomid DNA barcodes, we demonstrate here the use of this chironomid COI barcode database to improve the identification of DNA barcodes of prey in the liquefied diets of water mites. The species identifications of the COI barcodes of chironomids ingested by Lebertia davidcooki and L. quinquemaculosa are more diverse for L. davidcooki and include Parachironomus abortivus, Cryptochironomus ponderosus. Parachironomus tenuicaudatus, Glyptotendipes senilis, Dicrotendipes modestus, Chironomus riparius, Chironomus entis/plumosus, Chironomus maturus, Chironomus crassicaudatus, Endochironomus subtendens, Cricotopus sylvestris, Cricotopus festivellus, Orthocladius obumbratus, Tanypus punctipennis, Rheotanytarsus exiguus gr., and Paratanytarsus nr. bituberculatus.