The phylogeny and taxonomy of freshwater mussels of the genus Castalia in South America is complicated by issues of morphological plasticity and limited molecular genetic data. We present field data on the distributions of the nominal Castalia ambigua and C. inflata in the upper Paraguay River basin in Brazil based on original occurrence data at 23 sample sites and on historical records. The upper basin has distinct highland and lowland regions, the latter including the Pantanal wetland, where “C. ambigua” occurs in the highlands and “C. inflata” occurs in both regions. At Baixo Stream in the highlands, we observed individuals with shell morphologies of either C. ambigua or C. inflata, and also individuals with intermediate shell morphology. DNA sequence variation in the upland Baixo Stream and two representative lowland populations were screened. Two mitochondrial and three nuclear genes were sequenced to test hypotheses regarding the number of species-level phylogenetic lineages present. Reported individual DNA sequences from Amazon-basin C. ambigua and other Castalia and outgroup species were included in the analysis as outgroups. Individuals from the Paraguay River basin exhibited 17 haplotypes at the mitochondrial cytochrome oxidase I (COI) gene and nine at mitochondrial 16S rRNA. Analysis of haplotype networks and phylogenetic trees of combined COI + 16S rRNA sequences among individuals with the respective shell morphologies supported the hypothesis that C. ambigua and C. inflata from the Paraguay River basin belong to the same species and one phylogenetic lineage. No variation was observed at the nuclear 18S rRNA internal transcribed spacer, 28S rRNA, or H3NR histone genes among individuals used in this study. Across all markers, less variation was observed among Paraguay basin populations than between Paraguay and Amazon basin populations. Our results collectively suggest that: (1) “C. ambigua”, “C. inflata”, and morphologically intermediate individuals within the upper Paraguay drainage represent one phylogenetic lineage, (2) a phylogeographic divide exists between Castalia populations occurring in the Paraguay and Amazon River basins, and (3) the evolutionary and taxonomic uncertainties that we have identified among Castalia species should be thoroughly assessed across their distribution using both morphological and molecular characters.

We assessed parentage within and among maternity colonies of northern long-eared bats (Myotis septentrionalis (Trouessart, 1897)) in north-central Kentucky, USA, from 2011 to 2013 to examine colony social structure, formation, and membership dynamics. We intensively sampled colonies in close and remote (>10 km) proximity before and after targeted day-roost removal. Colonies were not necessarily composed of closely related individuals, although natal philopatry was common. Adjacent colonies often contained maternally related individuals, indicating that some pups did disperse, albeit not far from their natal home range. Whereas some young had been sired by males also collected on site, most had not, as would be expected since the species mates in fall near hibernacula across a wider landscape. The number of parentages that we inferred among colonies, however, suggests that outside the maternity season, social groups may be relatively flexible and open. Analysis of microsatellite DNA data showed a low FST (0.011) and best fit to a model of one multilocus genotypic cluster across the study area. We observed high turnover in colony membership between years in all colonies, regardless of roost-removal treatment. Our results suggest that female northern long-eared bats exhibit fidelity to a general geographic area and complex, dynamic social–genetic structure.

The Green River in Kentucky in the eastern United States is a freshwater mussel biodiversity hotspot, with 71 known species. Among them, the endangered Pleurobema plenum coexists with other morphologically similar species in the genera Fusconaia and Pleurobema, known colloquially as “pigtoes.” Identification of species in these genera is challenging even for mussel experts familiar with them. In our study, the correct identification of these species by experts ranged from 57% to 83%. We delineated taxonomic boundaries among seven species and tested for cryptic biodiversity among these look‐alike mussels utilizing mitochondrial and nuclear DNA sequence variation. Phylogenetic analysis of combined (1215 bp) mitochondrial DNA cytochrome oxidase I (COI) and NADH dehydrogenase 1 (ND1) genes showed five well‐diverged groups that included F. flava, F. subrotunda, P. cordatum, and P. plenum as distinct clades, with P. sintoxia and P. rubrum grouped into a single clade. While our mitochondrial DNA analyses did not distinguish P. sintoxia and P. rubrum as phylogenetically distinct species, the typical shell forms of these two nominal taxa are very distinct. Further phylogenetic analysis using nuclear ribosomal transcribed spacer region subunit I (ITS1) DNA sequences also showed that P. sintoxia and P. rubrum were not distinct lineages. No cryptic species were detected in the Fusconaia and Pleurobema samples analyzed from the Green River. The highest haplotype diversity (h), average number of nucleotide differences (k), and nucleotide diversity (π) were observed for F. subrotunda at both the COI (h = 0.896, k = 3.805, π = 0.00808) and ND1 (h = 0.984, k = 6.595, π = 0.00886) markers, with similarly high genetic diversity in the other taxa. Our results give managers confidence that cryptic taxa do not occur within or among these morphologically similar species in the Green River, and populations appear genetically diverse, indicative of large and healthy populations.