Intercontinental distributions, phylogenetic position and life cycles of species of Apharyngostrigea (Digenea, Diplostomoidea) illuminated with morphological, experimental, molecular and genomic data.

When subjected to molecular study, species of digeneans believed to be cosmopolitan are usually found to consist of complexes of species with narrower distributions. We present molecular and morphological evidence of transcontinental distributions in two species of Apharyngostrigea Ciurea, 1924, based on samples from Africa and the Americas. Sequences of cytochrome c oxidase I and, in some samples, internal transcribed spacer, revealed Apharyngostrigea pipientis (Faust, 1918) in Tanzania (first known African record), Argentina, Brazil, USA and Canada. Sequences from A. pipientis also match previously published sequences identified as Apharyngostrigea cornu (Zeder, 1800) originating in Mexico. Hosts of A. pipientis surveyed include definitive hosts from the Afrotropic, Neotropic and Nearctic, as well as first and second intermediate hosts from the Americas, including the type host and type region. In addition, metacercariae of A. pipientis were obtained from experimentally infected Poecilia reticulata, the first known record of this parasite in a non-amphibian second intermediate host. Variation in cytochrome c oxidase I haplotypes in A. pipientis is consistent with a long established, wide-ranging species with moderate genetic structure among Nearctic, Neotropic and Afrotropic regions. We attribute this to natural dispersal by birds and find no evidence of anthropogenic introductions of exotic host species. Sequences of CO1 and ITS from adult Apharyngostrigea simplex (Johnston, 1904) from Egretta thula in Argentina matched published data from cercariae from Biomphalaria straminea from Brazil and metacercariae from Cnesterodon decemmaculatus in Argentina, consistent with previous morphological and life-cycle studies reporting this parasite-originally described in Australia-in South America. Analyses of the mitochondrial genome and rDNA operon from A. pipientis support prior phylogenies based on shorter markers showing the Strigeidae Railliet, 1919 to be polyphyletic.

Molecular, morphological and experimental-infection studies of cercariae of five species in the superfamily Diplostomoidea (Trematoda: Digenea) infecting Biomphalaria straminea (Mollusca: Planorbidae) in Brazil.

Trematodes belonging to the superfamily Diplostomoidea have complex life cycles involving birds, mammals and reptiles as definitive hosts, and gastropods and different groups of invertebrates and vertebrates as intermediate hosts. Molecular studies of these parasites are numerous, but data from larval stages in molluscs remain scarce, particularly in South America. The present study focused mainly on five morphotypes of longifurcate cercariae found in Biomphalaria straminea (Dunker, 1848) from Belo Horizonte, Minas Gerais, Brazil, collected between 2009 and 2017. In each morphotype, nuclear internal transcribed spacer (ITS1-5.8S ITS-2) rDNA and mitochondrial cytochrome c oxidase 1 (COI) genes were sequenced. Laboratory-reared fish, Poecilia reticulata Peters, 1859 or snails, Biomphalaria glabrata (Say, 1818) were exposed to cercariae to obtain metacercariae. The morphology of cercariae, experimentally obtained metacercariae, and phylogenetic analyses led to the identification of three species of Diplostomidae [Austrodiplostomum compactum (Lutz, 1928), Crassiphialinae gen. sp. and Hysteromorpha sp.] and two species of Strigeidae (Cotylurus sp., Apharyngostrigea sp.). Previously published sequences allowed species-level identification for only A. compactum, although provisional identifications were possible in two cases. First, the COI from cercariae of Apharyngostrigea sp. in Brazil matched those of metacercariae from naturally infected Cnesterodon decemmaculatus (Jenyns, 1842) in Argentina; although a positive identification is not possible, the material presents morphological similarities with larval stages previously described for A. simplex. Secondly, Cotylurus sp. resembles C. lutzi. Our analysis of previously published COI sequences suggests that Cotylurus cornutus (Rudolphi, 1808) has a Holarctic distribution. Both the morphology of experimentally obtained metacercariae and COI sequences indicate that Hysteromorpha sp. in Brazil is distinct from congeners in North America [Hysteromorpha corti (Hughes, 1929)] and Europe [Hysteromorpha triloba (Rudolphi, 1819)].