The phylum Platyhelminthes is a diverse group of flatworms that includes parasites with serious impacts on human health, animal husbandry, aquaculture and wildlife management. Here we present degenerate primers for the barcode region of the mitochondrial cytochrome c oxidase I (COI) gene in flatworms. Although amplicons were obtained from a wide taxonomic range in the Cestoda and Trematoda, COI fragments from many taxa in these classes did not amplify. Primers specific to trematodes in the family Diplostomidae were also developed. Amplification success was much higher with diplostomid-specific primers and sequences were obtained from 504 of 585 specimens of Diplostomum and Tylodelphys. Sequences from the barcode region resolved all specimens to the species level, with mean divergence between congeners of 19% (3.9-25%). Because many of our specimens were small, we initially amplified part of the nuclear small subunit ribosomal (r) RNA gene to evaluate the quality and quantity of DNA in our specimens. Short sequences (~380 nt) of this gene were recovered from most specimens and can be used to distinguish specimens at the family level and often the generic level. We suggest that rRNA genes could be used to screen samples of completely unknown taxonomy, after which specific COI primers could be used to obtain species-level identifications.

Diplostomoid metacercariae parasitize freshwater fishes worldwide and cannot be identified to species based on morphology. In this study, sequences of the barcode region of cytochrome c oxidase subunit 1 (CO1) were used to discriminate species in 1088 diplostomoids, most of which were metacercariae from fish collected in the St. Lawrence River, Canada. Forty-seven diplostomoid species were detected, representing a large increase in known diversity. Most species suggested by CO1 sequences were supported by sequences of internal transcribed spacer (ITS) of rDNA and host and tissue specificity. Three lines of evidence indicate that physiological incompatibility between host and parasite is a more important determinant of host specificity than ecological separation of hosts and parasites in this important group of freshwater fish pathogens. First, nearly all diplostomoid species residing outside the lens of the eyes of fish are highly host specific, while all species that occur inside the lens are generalists. This can be plausibly explained by a physiological mechanism, namely the lack of an effective immune response in the lens. Second, the distribution of diplostomoid species among fish taxa reflected the phylogenetic relationships of host species rather than their ecological similarities. Third, the same patterns of host specificity were observed in separate, ecologically distinctive fish communities.

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The systematics of Diplostomum species, common intestinal parasites of piscivorous birds, has long been problematic, owing to phenotypic plasticity and the paucity of morphological features that are often subject to age-and host-induced variation. We sequenced the ITS1-5.8S-ITS2 regions of the rDNA from adult Diplostomum huronense, Diplostomum indistinctum, and Diplostomum baeri obtained from experimentally infected ring-bill gulls (Larus delawarensis) and compared them with partial ITS1 sequences from several species of Diplostomum in GenBank. The three North American species were distinguishable on the basis of ITS sequences. Sequences from D. huronense differed from those of D. indistinctum at 12 sites in ITS1 and 4 sites in ITS2, supporting morphological and morphometric data that indicate the two are distinct species. Sequences of D. huronense and D. indistinctum differed from those of D. baeri at 27 and 24 sites, respectively, in ITS1 and 15 and 12 sites, respectively, in ITS2. Phylogenetic analysis of partial ITS1 sequences revealed that the North American and European species of Diplostomum formed separate groups, with the former being basal to the latter. The results indicated that D. huronense and D. indistinctum from North America are distinct from Diplostomum spathaceum and other similar species from Europe. Furthermore, sequences from specimens identified as D. baeri from North America differed from those of D. baeri from Europe by 3.8% in ITS1 (23 sites). While morphologically similar, the two are not conspecific. Sequences of the North American species have been deposited in GenBank (AY 123042-123044).Résumé : La systématique des espèces de Diplostomum, parasites intestinaux communs des oiseaux piscivores, a toujours été problématique à cause de leur plasticité phénotypique et de la rareté des caractères morphologiques qui sont souvent soumis à des variations dues à l'âge et à l'hôte. Nous avons procédé au séquençage des régions ITS1-5.8S-ITS2 du gène d'ADNr de Diplostomum huronense, de Diplostomum indistinctum et Diplostomum baeri adultes provenant de goélands à bec cerclé (Larus delawarensis) infectés expérimentalement et nous avons comparé les séquences obtenues à des séquences partielles d'ITS1 de plusieurs espèces de Diplostomum de la banque génétique GenBank. Les trois espèces nord-américaines se distinguent par les séquences de leurs ITS. Les séquences de D. huronense diffèrent de celles de D. indistinctum à 12 sites sur ITS1 et à 4 sites sur ITS2, ce qui corrobore les données morphologiques et morphométriques qui indiquent qu'il s'agit de deux espèces distinctes. Les séquences de D. huronense et de D. indistinctum diffèrent de celles de D. baeri à 27 et 24 sites sur ITS1 et à 15 et 12 sites sur ITS2, respectivement. L'analyse phylogénétique des séquences partielles D'ITS1 a révélé que les espèces nord-américaines et européennes de Diplostomum forment deux groupes distincts, le premier groupe étant plus ancien. Les résultats indiquent que D. huronense et D. indistinctum d'Amérique du Nord so...

Given that numerous amphibians are suffering population declines, it is becoming increasingly important to examine the relationship between disease and environmental disturbance. Indeed, while many studies relate anthropogenic activity to changes in the parasitism of snails and fishes, little is known of the impact on the parasites of amphibians, particularly from agriculture. For 2 years, the parasite communities of metamorphic northern leopard frogs from 7 agricultural wetlands were compared with those from 2 reference wetlands to study differences in parasite community diversity and abundance of various species under pristine conditions and 3 categories of disturbance: only agricultural landscape, only pesticides, and agricultural landscape with pesticides. Agricultural (and urban) area was negatively related to species richness, and associated with the near absence of adult parasites and species that infect birds or mammals. We suggest that agriculture and urbanization may hinder parasite transmission to frogs by limiting access of other vertebrate hosts of their parasites to wetlands. The only parasite found at all localities was an unidentified echinostome infecting the kidneys. This parasite dominated communities in localities surrounded by the most agricultural land, suggesting generalist parasites may persist in disrupted habitats. Community composition was associated with dissolved organic carbon and conductivity, but few links were found with pesticides. Pollution effects may be masked by a strong impact of land use on parasite transmission.

Parasite communities tend to be dissimilar in hosts that are geographically, phylogenetically, ecologically and developmentally distant from one another. The decay of community similarity is a powerful and increasingly common method of studying parasite beta diversity, but most studies have examined only a single type of distance. Here, we evaluate distances based on the phylogeny, ecology, spatial proximity and size of hosts, as predictors of the similarity of parasite communities in individual hosts, host populations and host species. We surveyed parasites in six species of fish collected simultaneously from six localities in the St. Lawrence River, Canada, and species in a common group of larval parasites were discriminated using DNA sequences from barcode region of cytochrome c oxidase I. Distances based on the habitat use patterns of host species were good predictors of short‐term, ecological similarity of parasite communities, such as that operating at the scale of the individual host. The genetic distance between host species was associated with almost all types of similarity at all scales, particularly qualitative and phylogenetic similarity of parasite communities at the level of populations and meta‐populations of hosts. The trophic level, diet, spatial proximity and size of hosts were poor predictors of parasite community similarity. The increased taxonomic resolution provided by molecular data increased the explanatory power of regression models, and different factors were implicated when parasite species were distinguished with DNA barcodes than when larval parasites were lumped into morphospecies, as is commonly practiced.

Because the taxonomy of trematodes is based on adults, the larval stages of most digeneans cannot be identified to species based on morphology alone. Molecular data provide a means of linking larval stages to known adults. We obtained sequences from the barcode region of cytochrome oxidase I (CO1) from adult and larval parasites of fish, frogs, birds, and mammals across North America. Sequences from adult Apharyngostrigea cornu, Hysteromorpha triloba, and Alaria mustelae (Diplostomoidea: Digenea) from definitive hosts matched those of meta- and mesocercariae from fish and frogs. These data provided new information on the distributions of all 3 parasite species. Metacercariae of A. cornu, which have not been previously reported in North American hosts, were found in Notemigonus crysoleucas, Pimephales notatus, and Catostomus commersonii in the St. Lawrence River. Metacercariae of H. triloba are reported in Canadian waters and in N. hudsonius for the first time. Alaria mustelae is reported for the first time in frogs from Quebec, Canada, and an additional species of Alaria was detected in California. Sequences of internal transcribed spacer rDNA from a subset of specimens support the same species boundaries indicated by CO1 divergence. There was little divergence in CO1 sequences from an unidentified diplostomid species sampled at a large spatial scale.