RESUMO
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.
Assuntos
Trematódeos , Infecções por Trematódeos , Animais , Brasil , Genômica , Estágios do Ciclo de Vida , Filogenia , Trematódeos/genética , Infecções por Trematódeos/veterináriaRESUMO
Zygocotyle lunata inhabits the caecum of birds and mammals from the American continent. This amphistome parasite is easily maintained in the laboratory and serves as a model organism in life-cycle studies, but it has seldom been studied using molecular data. Neither the position of Z. lunata in the superfamily Paramphistomoidea nor the monophyly of the Zygocotylidae has been evaluated with molecular phylogenetic methods. In the present study, adult specimens of Z. lunata obtained experimentally in mice from Brazil were submitted to molecular studies. Partial sequences of nuclear (1261 bp of 28S and 418 bp of 5.8S-ITS-2) and mitochondrial (1410 bp of cytochrome c oxidase 1, cox1) markers were compared with published data. In the most well-resolved phylogeny, based on 28S sequences, Z. lunata clustered in a well-supported clade with Wardius zibethicus, the only other species currently included in the Zygocotylidae, thus confirming the validity of this family. Divergence of 28S sequences between these species was 2.2%, which falls in the range of intergeneric variation (0.9-5.6%) observed in the other two monophyletic groups in the 28S tree, i.e., representatives of Gastrodicidae and Neotropical cladorchiids (Cladorchiidae). Analysis of ITS-2 and two parts of the cox1 gene placed Z. lunata within poorly resolved clades or large polytomies composed of several paramphistomoid families, without clarifying higher-level phylogenetic relationships. The cox1 of a Brazilian isolate of Z. lunata is 99.6% similar to a Canadian isolate, confirming the pan-American distribution of the species. Finally, our phylogenetic reconstructions of Paramphistomoidea revealed a complex scenario in the taxonomic composition of some amphistome families, which highlights a need for further integrative studies that will likely result in rearrangements of traditional morphology-based classifications.
Assuntos
Doenças das Aves/parasitologia , Ceco/parasitologia , Paramphistomatidae/genética , Paramphistomatidae/isolamento & purificação , Filogenia , Infecções por Trematódeos/veterinária , Animais , Aves/parasitologia , Brasil , Canadá , Feminino , Estágios do Ciclo de Vida , Masculino , Camundongos , Paramphistomatidae/classificação , Paramphistomatidae/crescimento & desenvolvimento , Infecções por Trematódeos/parasitologiaRESUMO
Diplostomum ardeae Dubois, 1969 has seldom been reported since its description from the great blue heron (Ardea herodias L., 1758) in the USA. Sequences obtained in this study from the barcode region of cytochrome c oxidase 1 (CO1) in diplostomids from black-crowned night heron (Nycticorax nycticorax (L., 1758)) in Puerto Rico matched data from D. ardeae from A. herodias in the type region. We also obtained DNA barcodes from morphologically similar diplostomids from a rufescent tiger heron (Tigrisoma lineatum (Boddaert, 1783)) and from metacercariae from eye lenses of Trachelyopterus galeatus (Linnaeus, 1766) from the Paraná River basin in Argentina and Brazil, respectively. Barcodes matched (97-100% identity) in these South American adult and larval specimens as well as in recently published sequences from metacercariae from 11 other siluriform fishes from the same region. Barcodes from the South American species, which we describe as Diplostomum lunaschiae n. sp., differed from those of D. ardeae by 7.2-9.8%, and the new species differs from D. ardeae in its size, pharynx:oral sucker length ratio, egg:body length ratio, and distribution of vitellaria. As in prior phylogenetic analysis of CO1 sequences, both D. ardeae and D. lunaschiae n. sp. were not associated with Diplostomum. In more character-rich analyses of nuclear rDNA and of mitochondrial genomes, D. ardeae was an early divergent member of clades of species of Diplostomum. Consequently, we continue to consider D. ardeae and D. lunaschiae n. sp. members of Diplostomum, in contrast to recent suggestions that these species may belong to a different genus.
Assuntos
Aves/parasitologia , Estágios do Ciclo de Vida , Filogenia , Trematódeos/classificação , Trematódeos/crescimento & desenvolvimento , Animais , Peixes-Gato/parasitologia , Código de Barras de DNA Taxonômico , DNA Ribossômico/genética , Genoma Mitocondrial/genética , Metacercárias/classificação , Metacercárias/genética , Porto Rico , Trematódeos/anatomia & histologia , Trematódeos/genéticaRESUMO
Metacercariae of Clinostomum Leidy, 1856 are frequently encountered in freshwater fish. In 2015, a provisional species of Clinostomum in People's Republic of China (PRC) was distinguished from C. complanatum (Rudolphi, 1819) in Europe based on divergent cytochrome c oxidase I (CO1). However, in subsequent studies in East Asia, the same divergent CO1 genotype was identified as C. complanatum. These matching sequences suggest that either the provisional East Asian species was incorrectly distinguished from C. complanatum in 2015 or that C. complanatum in East Asia was misidentified in later studies. We tested these alternatives by sequencing the mitochondrial genome of C. complanatum in Italy, which was 5.7% divergent from a previously published sequence from Clinostomum in PRC, including differences in 80 of 3390 (2.4%) translated amino acids. Partial CO1 sequences of specimens from PRC and those from Italy, Romania, and Turkey also each formed reciprocally monophyletic clades. Partial CO1 from the East Asian clade varied by mean 3.6% (range 2.4-4.8%) from C. complanatum from Italy, Romania, and Turkey; mean intra-clade CO1 variation was 0.3% (range 0-1.9%). Metacercariae from Europe and East Asia display significant morphometric variation, and data from the literature suggest morphological differences in the genital complex of adults. Although sequences of nuclear rDNA did not differ between isolates from the west and East Asia, taken together, these results lead us to describe a new species of Clinostomum.
Assuntos
Filogenia , Trematódeos/classificação , Animais , Ásia , DNA Ribossômico/genética , Peixes/parasitologia , Água Doce/parasitologia , Genoma Mitocondrial/genética , Metacercárias/anatomia & histologia , Metacercárias/classificação , Metacercárias/genética , Especificidade da Espécie , Trematódeos/anatomia & histologia , Trematódeos/genéticaRESUMO
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)].
Assuntos
Biomphalaria/parasitologia , Cercárias/anatomia & histologia , Trematódeos/anatomia & histologia , Infecções por Trematódeos/parasitologia , Animais , Brasil , Feminino , Masculino , Trematódeos/classificação , Trematódeos/genéticaRESUMO
Postharmostomum commutatum (Dietz, 1858), a parasite of the caeca of poultry, has been reported from many different parts of the world. Despite its importance, there are no molecular sequences available and its phylogenetic position is unknown in relation to other members of Brachylaimoidea, a group in which taxonomic confusion reigns. Here, morphological and molecular techniques were used to study digeneans from the caeca of free-range chickens found naturally infected in the municipality of Viçosa, state of Minas Gerais, Brazil, between August 2017 and May 2018. The specimens were identified as P. commutatum, with Postharmostomum gallinum Witenberg, 1923 herein considered a junior synonym. Sequences obtained for the 28S, ITS2, and cox-1 genes were compared with sequences available from other species of Brachylaimoidea. Phylogenetic analysis of the three markers indicates P. commutatum formed an isolated lineage from other brachylaimoids, supporting the distinct status of the genus. The topology of phylogenetic trees obtained suggests that the morphology-based classification of families of Brachylaimoidea is artificial and new rearrangements of some genera or creation of new families may be necessary. The sequences newly obtained here will be useful for testing the cosmopolitan distribution of P. commutatum.
Assuntos
Ceco/parasitologia , Galinhas/parasitologia , Trematódeos/classificação , Trematódeos/genética , Animais , Brasil , Ciclo-Oxigenase 1/genética , DNA de Helmintos/genética , DNA Intergênico/genética , Filogenia , Aves Domésticas/parasitologia , RNA Ribossômico 28S/genéticaRESUMO
Species of the genus Posthodiplostomum are intestinal parasites of piscivorous birds with worldwide distribution. Recent molecular surveys have focused on developmental stages from vertebrate hosts, with few sequences from larvae found in molluscs. Moreover, most published molecular data are from collections in North America, Europe and Asia, and there are no data for South American species. In the present study, cercariae found in the freshwater limpet Gundlachia ticaga from Brazil were used for morphological, experimental and molecular studies. The longifurcate cercariae, Neascus-type metacercariae obtained in experimentally infected guppies (Poecilia reticulata), and an adult parasite recovered from a mouse were morphologically identified as Posthodiplostomum nanum. Sequences of DNA from internal transcribed spacer (ITS) rDNA and cytochrome c oxidase 1 (cox1) from the cercariae and metacercariae were compared with published data, yielding no matches (ITS divergenceâ¯≥â¯5.5%, cox1â¯≥â¯13.9%). Phylogenetic analyses of the ITS sequences suggest paraphyly within the genus Posthodiplostomum, or misidentifications of parasites in molecular studies. For example, ITS sequences of Mesoophorodiplostomum pricei and Posthodiplostomum centrarchi or the unnamed species Posthodiplostomum sp. 8 diverge by only 1.1-1.2% and 0.6%, respectively, and all three species fall within a well-supported clade, suggesting that these isolates are congeneric. The phylogenetic tree obtained for cox1 sequences, although not well resolved, reveals that the type species of the genus, Posthodiplostomum cuticola, does not group with any species previously identified as Posthodiplostomum. Overall, the analyses of molecular data here obtained for P. nanum compared with sequences of related species suggest that a review of this group is necessary. Such studies may result in a split of the genus Posthodiplostomum and the transference of some species currently assigned in this genus to other already described genera.