Phylogenetic relationships, expanded diversity and distribution of Crassiphiala spp. (Digenea, Diplostomidae), agents of black spot disease in fish.

Crassiphiala is a monotypic genus of diplostomid digeneans and is the type genus of the subfamily Crassiphialinae. The type species Crassiphiala bulboglossa parasitizes kingfishers in the Nearctic and has a Neascus-type metacercaria that encysts on fish intermediate hosts, often causing black spot disease. While recent molecular phylogenetic studies included some members of the Crassiphialinae, no DNA sequence data of Crassiphiala is currently available. Our molecular and morphological study of adult and larval crassiphialines from the Americas revealed the presence of at least three lineages of Crassiphiala from the Nearctic and two lineages from the Neotropics. This is the first record of Crassiphiala from the Neotropics. Herein, we provide the first molecular phylogeny of the Diplostomoidea that includes Crassiphiala. Our data revealed 0.2-2.4% divergence among 28S sequences and 11-19.8% among CO1 sequences of lineages of Crassiphiala. The results of our analyses did not support the monophyly of Crassiphialinae. Our results clearly demonstrated that the diversity of Crassiphiala has been underestimated.

Phenotypic consequences of genetic variation in a gynogenetic complex of Phoxinus eos-neogaeus clonal fish (Pisces: Cyprinidae) inhabiting a heterogeneous environment.

We examined the genetic composition, habitat use, and morphological variation of a Phoxinus eos-neogaeus unisexual hybrid complex and its sexually reproducing progenitor species inhabiting beaver-modified drainages of Voyageurs National Park, Minnesota. In addition to the single diploid P. eos-neogaeus gynogenetic clone, triploid and diploid-triploid mosaic biotypes were present at our study sites. Both P. eos and P. neogaeus, and all three hybrid biotypes were ubiquitous throughout one intensively surveyed drainage, but abundances and relative frequencies of the parental species and hybrids varied considerably within and among successional environments. Data from a large number of additional sites indicated that the proportion of polyploid hybrids within an environment was negatively related to hybrid relative frequency, implying that the genomic constitution of hybrids is an important determinant of clonal fitness among successional environments. Statistical comparisons of variation along size-free multivariate body shape axes indicated that despite its genetic uniformity, the P. eos-neogaeus clone is no less variable than its sexual progenitors, suggesting that a single genotype may actually respond to environmental variation with as much phenotypic variation as a genetically variable sexual population. The incorporation and expression of a third genome in triploid and diploid-triploid mosaic biotypes derived from the gynogenetic clone significantly expanded phenotypic variation of the clone. This additional variation results in greater similarities in habitat use and morphological overlap with the parental species, primarily P. eos, the predominant sperm donor for gynogenetic hybrid females in this complex. Polyploid augmentation of a diploid gynogenetic clone appears to be typical in the P. eos-neogaeus complex, and the additional genetic and phenotypic variation that it generates has potentially significant ecological and evolutionary consequences for the success and persistence of a single genotype in highly variable environments.

Fish recruitment, dispersal, and trophic interactions in a heterogeneous lotic environment.

I examine how dispersal of juvenile creek chubs (Semotilusatromaculatus) from beaver ponds into adjacent stream environments interacts with temporal abiotic variability to influence fish foraging, growth, and long-term persistence in the lotic ecosystem. Minnow trapping in upstream and downstream beaver ponds, along with weir traps used to monitor directional movement, indicated that most chubs colonized the stream from the downstream beaver pond. Large annual fluctuations in density of age 0 creek chubs occurred in the stream over a 10-year sampling period. Multiple regression analysis indicated that stream temperature, precipitation, and the density of reproductive creek chubs were not correlated with summer density of age 0 chubs in the stream. The factor most strongly associated with increased density of age 0 creek chubs was creation of the downstream beaver pond during the 6th-7th years of the study, suggesting dispersal from the pond was the primary factor determining age 0 fish density in the stream. Most individuals in the strong year classes neither persisted in the stream through their first winter nor resulted in an increased abundance of older age classes in later years. Comparison of age 0 fish density in summer to the proportion of fish surviving to age 1 in spring suggested that overwinter mortality was density dependent. Furthermore, a comparison of the size structure for age 0 individuals in summer to age 1 individuals the following spring indicated that winter mortality was size dependent. Experiments in an artificial stream adjacent to the natural channel revealed that fish growth was strongly density dependent, decreasing as fish density increased across both spring and summer, and elevated and low discharge. The decline in invertebrate prey captured by the fish and the subsequent decline in fish growth appeared to be particularly pronounced under low discharge in summer. Changes in juvenile creek chub density had no significant effect on benthic insect or crustacean abundance, suggesting that exploitative competition for limited invertebrate drift resources was a more important cause of density- dependent growth than depressed local benthic invertebrate abundance. These results suggest that lotic regions adjacent to beaver ponds act as potential reproductive "sinks" for dispersing juveniles confronting seasonal and flow-mediated restrictions on resource acquisition and growth, and the occurrence of seasonal bottlenecks to their survival, especially harsh winter conditions.

The origin of Phoxinus eos-neogaeus unisexual hybrids.

Phoxinus eos-neogaeus unisexual hybrids (Cyprinidae, Pisces) are among the few vertebrate taxa known to reproduce clonally by gynogenesis. These taxa have a broad distribution in North America, mostly located in regions previously covered by the last Pleistocene ice sheet. To assess whether asexual hybrids dispersed from glacial refuges at the end of the Pleistocene or they originated from current hybridization events, genetic diversity of mitochondrial DNA (mtDNA) sequences and microsatellite loci was determined in populations from 16 different sites in the Mississippi-Missouri River (Nebraska and Montana), Rainy River-Hudson Bay (Minnesota), and St Lawrence River (Quebec) drainages. The maternal species (P. neogaeus) occurred in Minnesota and Nebraska but was absent from Montana sites and was restricted to only two of 11 lakes sampled in Quebec, although hybrids were present at all sites. The genetic survey revealed a total of 49 clones, originating from 14 hybridization events. Several of the lineages were characterized by mtDNA haplotypes not detected in the maternal ancestor. Lineages as well as clones frequently displayed a large geographical distribution at a regional scale. Dating of hybridization events suggested a relatively recent origin (<50,000 years ago) from the Mississippi glacial refuge, even in regions not covered by the last Pleistocene glacier. Altogether, these results indicate P. eos-neogaeus hybrids are not the result of current hybridization events, but display a pattern predicted by postglacial dispersal. Our findings have considerable implications for the nature of selection processes affecting the diversity of these asexual taxa and their coexistence with sexual ancestors.