RT Journal Article T1 Novel Microsatellite Loci, Cross-Species Validation of Multiplex Assays, and By-Catch Mitochondrial Genomes on Ochthebius Beetles from Supratidal Rockpools A1 García Meseguer, Antonio José A1 Villastrigo, Adrián A1 Mirón Gatón, Juan María A1 Millán, Andrés A1 Velasco, Josefa A1 Muñoz Gabaldón, Irene AB Here we focus on designing, for the first time, microsatellite markers for evolutionary and ecological research on aquatic beetles from the genus Ochthebius (Coleoptera, Hydraenidae). Some of these non-model species, with high cryptic diversity, exclusively inhabit supratidal rockpools, extreme and highly dynamic habitats with important anthropogenic threats. We analysed 15 individuals of four species (O. lejolisii, O. subinteger, O. celatus, and O. quadricollis) across 10 localities from the Mediterranean coasts of Spain and Malta. Using next-generation sequencing technology, two libraries were constructed to interpret the species of the two subgenera present consistently (Ochthebius s. str., O. quadricollis; and Cobalius, the rest of the species). Finally, 20 markers (10 for each subgenus) were obtained and successfully tested by cross-validation in the four species under study. As a by-catch, we could retrieve the complete mitochondrial genomes of O. lejolisii, O. quadricollis, and O. subinteger. Interestingly, the mitochondrial genome of O. quadricollis exhibited high genetic variability compared to already published data. The novel SSR panels and mitochondrial genomes for Ochthebius will be valuable in future research on species identification, diversity, genetic structure, and population connectivity in highly dynamic and threatened habitats such as supratidal coastal rockpools. PB MDPI YR 2023 FD 2023-11-15 LK https://hdl.handle.net/20.500.14352/104023 UL https://hdl.handle.net/20.500.14352/104023 LA eng NO García-Meseguer AJ, Villastrigo A, Mirón-Gatón JM, Millán A, Velasco J, Muñoz I. Novel Microsatellite Loci, Cross-Species Validation of Multiplex Assays, and By-Catch Mitochondrial Genomes on Ochthebius Beetles from Supratidal Rockpools. Insects. 2023 Nov 15;14(11):881. doi: 10.3390/insects14110881. PMID: 37999080; PMCID: PMC10672297. NO 2022 Descuento UCM NO Ministerio de Ciencia, Innovación y Universidades NO European Commission DS Docta Complutense RD 11 abr 2025