Several nest-building North American minnows (Cyprinidae) function as reproductive hosts to nest associatesÀspecies that require nests of other species for spawning. Understanding the microhabitat preferences of hosts can yield insight into the reproductive ecology of many species, especially of nest associates that can utilize nests of two or more hosts. We observed nests of Central stoneroller Campostoma anomalum in which several associate species were actively spawning. Bluehead chubs Nocomis leptocephalus began constructing nests two days later in the same stream, at which time associates abandoned stoneroller nests and continued on to spawn on chub nests. This presented a unique opportunity for accomplishing two objectives: (1) quantifying stoneroller nesting microhabitat preference and (2) comparing stoneroller and chub habitat preference to gain insight into the mechanisms that may drive host switching by nest associates. We measured substrate size, current velocity, water depth, and egg depth on seven paired stoneroller and chub nests, and compared these measurements to paired microhabitat measurements at a randomly selected point near each nest. Repeated measures analysis of variance with post hoc Tukey tests revealed that stonerollers exhibited distinct nesting microhabitat preferences from chubs. Gravel on stoneroller nests was considerably smaller than on chub nests and stonerollers nested in shallower depths than chubs. However, both species nested at similar current velocities. If nest associates switch partners based on the physical characteristics of nests, then substrate size is likely the most important factor. The larger gravel sizes on chub nests likely provide better egg aeration than stoneroller nests. Chub nests may also be safer for associate broods because male Bluehead chubs cover eggs with gravel after spawning; stonerollers do not. Future work should take an experimental approach to elucidate these mechanisms.

Larval fish ecology is poorly characterized because sampling is difficult and tools for phenotypically identifying larvae are poorly developed. While DNA barcoding can help address the latter problem, ‘universal’ primers do not work for all fish species. The Roanoke River in the southeastern United States includes seven darters (Family Percide: Tribe Etheostomatini). We made 393 collections of larval fishes in 2015 and 2018, examined darter larvae for morphometric and pigmentation traits, developed PCR primers amplifying darter DNA, and evaluated three gear types for collecting larval darters. Amplified DNA sequences for 1351 larvae matched archived mitochondrial cytochrome oxidase I sequences for darters occurring in the ecosystem. Larval darters were classified to genus with 100% accuracy using the ratio of pectoral fin length to body length; however, identification to species using morphometrics alone was subject to a misclassification rate of 11.8%, which can be resolved by considering pigmentation patterns. Gear-types varied considerably in their capture efficacy for larval darters; most Percina larvae were collected in drift nets. Larval Percina species appeared in the drift before Etheostoma species in both study years. Application of molecular genetic and phenotypic tools to larval fish identification can advance understanding of larval darter ecology.

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