Phylogenetic Relationships of Avian Cestodes from Brine Shrimp and Congruence with Larval Morphology.

Determining molecular markers for parasites provides a useful tool for their identification, particularly for larval stages with few distinguishable diagnostic characters. Avian cestodes play a key role in the food webs and biodiversity of hypersaline wetlands, yet they remain understudied. Using naturally infected Artemia, we identified cestode larvae (cysticercoids), assessed their genetic diversity, and explored phylogenetic relationships in relation to larval morphology and waterbird final hosts. We obtained partial 18S rDNA sequences for 60 cysticercoids of the family Hymenolepidae infecting Artemia spp. from seven localities and three countries (Spain, the USA, and Chile). We present the first DNA sequences for six taxa: Confluaria podicipina, Fimbriarioides sp., Flamingolepis liguloides, Flamingolepis sp. 1, Flamingolepis sp. 2, and Hymenolepis californicus. Intraspecific sequence variation (0.00-0.19% diversity) was lower than intergroup genetic distance (0.7-14.75%). Phylogenetic analysis revealed three main clades: 1-Flamingolepis, 2-Fimbriarioides, 3-Confluaria and Hymenolepis, all of which separated from hymenolepidids from mammals and terrestrial birds. This clear separation among taxa is congruent with previous morphological identification, validating the 18S gene as a useful marker to discriminate at generic/species level. Working with intermediate hosts allows the expansion of knowledge of taxonomic and genetic diversity of cestodes in wildlife, as well as elucidation of their life cycles.

Late Cenozoic history and the role of Beringia in assembling a Holarctic cestode species complex.

The dynamic climate history that drove sea level fluctuation during past glacial periods mediated the movement of organisms between Asia and North America via the Bering Land Bridge. Investigations of the biogeographic histories of small mammals and their parasites demonstrate facets of a complex history of episodic geographic colonization and refugial isolation that structured diversity across the Holarctic. We use a large multi-locus nuclear DNA sequence dataset to robustly resolve relationships within the cestode genus Arostrilepis (Cyclophyllidea: Hymenolepididae), a widespread parasite of predominantly arvicoline rodents (voles, lemmings). Using this phylogeny, we confirm that several Asian Arostrilepis lineages colonized North America during up to four distinct glacial periods in association with different rodent hosts, consistent with taxon-pulse dynamics. A previously inferred westward dispersal across the land bridge is rejected. We also refine interpretations of past host colonization, providing evidence for several distinct episodes of expanding host range, which probably contributed to diversification by Arostrilepis. Finally, Arostrilepis is shown to be paraphyletic with respect to Hymenandrya thomomyis, a parasite of pocket gophers, confirming that ancient Arostrilepis species colonized new host lineages upon arriving in North America.

Redescription and taxonomic position of Rodentolepis (sensu lato) asymmetrica (Janicki, 1904), with the erection of Kontrimavichusia n. g. (Eucestoda: Hymenolepididae) from arvicoline rodents (Rodentia: Cricetidae).

Rodentolepis (sensu lato) asymmetrica (Janicki, 1904) is redescribed on the basis of materials from Microtus agrestis, Microtus arvalis and Myodes glareolus from Eastern Europe (Belarus, Lithuania, Latvia, Estonia, Ukraine and the north-eastern part of Russia). A new genus, Kontrimavichusia n. g., is proposed for it based on morphological and molecular evidence. The unique morphological diagnostic characters of this currently monotypic genus include the presence of an armature on the suckers, ventral osmoregulatory canals connected with irregularly spaced transverse anastomoses, an internal seminal vesicle with circular musculature and uterus with numerous diverticula, situated dorsally to the genital ducts, extending bilaterally beyond the longitudinal osmoregulatory canals. In addition, the new genus differs from morphologically related genera of the Rodentolepis clade by the structure of its vagina. The copulatory part of the vagina in specimens of Kontrimavichusia n. g. is surrounded by circular musculature and covered externally by a dense layer of intensely-stained cells; the conductive part of the vagina is clearly distinguishable from the seminal receptacle. Molecular analysis of the partial 28S rRNA gene fully supports the erection of Kontrimavichusia n. g. as a distinct lineage.

Rodentolepis microstoma isolated from different species of Sigmodontinae rodents (Rodentia: Cricetidae) in the Cuenca del Plata, Argentina: Morphological aspects and molecular characterization.

The purpose of this paper was to study specimens of the genus Rodentolepis isolated from eight species of Sigmodontinae rodents (Rodentia: Cricetidae) from six provinces in the Cuenca del Plata, Argentina, based on morphological, morphometric and molecular characteristics (ITS1 rDNA and cox1 mtDNA). The genetic distances among studied specimens and other Hymenolepididae from rodents available in the GenBank were analyzed and phylogenetic inferences were provided. A total of 955 specimens of Sigmodontinae rodents were examined from seven localities of six provinces in the Cuenca del Plata region in Argentina. Tapeworms were removed from the rodents' small intestines. Conventional studies were used for the morphological and molecular analysis. Specimens of R. microstoma were identified. An amended diagnosis and detailed morphological description of this species is provided. The molecular analyses showed that the specimens studied form the same clade as that of R. microstoma previously studied from other hosts and regions. The genetic polymorphisms of R. microstoma observed corresponded to different groups of species hosts and regions. Moreover, eight species of sigmodontine rodents and 33 localities from the Cuenca del Plata region in Argentina constitute new host and geographical records. This study shows the importance of using integrative taxonomic approaches that combine morphological and molecular characters to understand biological diversity. Moreover, the discovery of R. microstoma in humans suggests the importance of further studies on this zoonotic cestode. This study provides important data on the taxonomy and distribution of R. microstoma to advance knowledge of the transmission dynamics of this parasite.

Hymenolepidid cestodes: Diversity, morphological and molecular characterization of a new species, and phylogeny of parasitic species of rodents from North and South America.

Rodents are hosts of a wide diversity of cestodes. Fifteen genera included in the family Hymenolepididae parasitize rodents, and only four of these genera have been recorded from the Neotropical region. The purpose of this paper is to update species of Hymenolepididae from rodents, describe a new species of Hymenolepis based on morphological and molecular characterization (ITS1 rDNA and cox1 mtDNA), comparing the features among the species from North and South American rodents, and provide phylogenetic inferences of Hymenolepididae from rodents based on sequences available in the GenBank. Rodents were collected in the Parque Provincial Ernesto Tornquist, Buenos Aires, Argentina. Hymenolepis ivanovae n. sp. differs from other Hymenolepis species registered from North and South American rodents by body size, scolex, suckers, cirrus sac, cirrus, testes, and eggs, among others. Comparative morphometric data for Hymenolepis species from North and South American rodents is provided. Molecular analyses place H. ivanovae n. sp. within the genus Hymenolepis with strong support, and show it close to species of zoonotic importance. The new species is the first species of Hymenolepis described from Sigmodontinae rodents.

Molecular characterisation and genetic affinities of Cyclophyllidean cestodes infecting wild rats in Peninsular Malaysia

@#Cestode infections is widely reported in rodents, however species identification remains problematic due to the genetic or interspecies variation. Therefore, this study was aimed to verify the Cyclophyllidean parasites recovered from wild rats captured from different forest types using molecular based methods. Maximum-likelihood (ML) and neighbour-joining (NJ) trees were constructed inferred from 18 small subunit nuclear ribosomal RNA gene (18SrDNA) and mitochondrial cytochrome c oxidase subunit one gene (COX1) sequences of cestode worms recovered from 124 individuals from four rat species. Sequences obtained from both Hymenolepis diminuta and Hydatigera parva represents the first records in Malaysia. All the sequences were successfully amplified with product with total length of 205 and 1202 base pairs (bp), respectively. Three cestode species from the Family Hymenolepididae (Hymenolepis diminuta) and Family Taeniidae (Hydatigera parva; Hydatigera taeniaeformis) were successfully characterized using phylogenetic analyses and haplotype networking. Phylogenetic analysis showed that H. diminuta, Hydatigera parva (Hy. parva) and Hydatigera taeniaeformis (Hy. taeniaeformis) formed its own monophyletic clade in 18SrDNA analyses. Results also showed that Hy. taeniaeformis shared the same haplotype group with Hy. taeniaeformis from China (COX1) and linked with Hy. taeniaeformis from Japan (18SrDNA) while the Malaysian H. diminuta clearly formed a separate haplotype and networked with other regions. The Malaysian Hy. parva isolation, on the other hand, appeared to be genetically distinct from the European Hy. parva (Spain) strain, but closely linked to the local isolates. Molecular methods employed successfully improved in the detection of complex species in this group. The findings showed that molecular data can be useful to deeply study intra-specific variation in other cestode worms.

Phylogeny of hymenolepidids (Cestoda: Cyclophyllidea) from mammals: sequences of 18S rRNA and COI genes confirm major clades revealed by the 28S rRNA analyses.

The aim of the study is to test a hypothesis for the phylogenetic relationships among mammalian hymenolepidid tapeworms, based on partial (D1-D3) nuclear 28S ribosomal RNA (rRNA) genes, by estimating new molecular phylogenies for the group based on partial mitochondrial cytochrome c oxidase I (COI) and nuclear 18S rRNA genes, as well as a combined analysis using all three genes. New sequences of COI and 18S rRNA genes were obtained for Coronacanthus integrus, C. magnihamatus, C. omissus, C. vassilevi, Ditestolepis diaphana, Lineolepis scutigera, Spasskylepis ovaluteri, Staphylocystis tiara, S. furcata, S. uncinata, Vaucherilepis trichophorus and Neoskrjabinolepis sp. The phylogenetic analyses confirmed the major clades identified by Haukisalmi et al. (Zoologica Scripta 39: 631-641, 2010): Ditestolepis clade, Hymenolepis clade, Rodentolepis clade and Arostrilepis clade. While the Ditestolepis clade is associated with soricids, the structure of the other three clades suggests multiple evolutionary events of host switching between shrews and rodents. Two of the present analyses (18S rRNA and COI genes) show that the basal relationships of the four mammalian clades are branching at the same polytomy with several hymenolepidids from birds (both terrestrial and aquatic). This may indicate a rapid radiation of the group, with multiple events of colonizations of mammalian hosts by avian parasites.

A new genus and two new species of unarmed hymenolepidid cestodes (Cestoda: Hymenolepididae) from geomyid rodents in Mexico and Costa Rica.

Two new cestodes of the family Hymenolepididae are described from two species of rodents of the family Geomyidae collected in Mexico and Costa Rica. One new species of Hymenolepis is described from Cratogeomys planiceps Merriam 1895 from near Toluca, Mexico and another that we allocate to a new genus is described from Heterogeomys heterodus (Peters, 1865) from near Irazú Volcano, Costa Rica. Hymenolepis s. str. includes those Hymenolepididae with an apical organ, with no hooks on suckers or apical organ, and three testes. Hobergia irazuensis n. gen., n. sp. includes a hymenolepidid with an apical organ, unarmed scolex, small pockets termed foveolae, in which the suckers completely retract, and extremely bi-lobed ovary. Multivariate morphometric analysis showed good separation of these species from all other hymenolepidids possessing an apical organ and lacking a well developed rostellum and rostellar hooks in the Nearctic and Neotropical regions.

Exploring parasites in extreme environments of high conservational importance: Artemia franciscana (Crustacea: Branchiopoda) as intermediate host of avian cestodes in Andean hypersaline lagoons from Salar de Atacama, Chile.

The hypersaline lagoons located in evaporation basins or salars (salt flats) in the Atacama Desert are extreme environments harbouring a specialised biota able to survive harsh conditions for life. The knowledge of the parasitic biodiversity of these extreme habitats is still scarce despite their functional importance in regulating relevant non-economic services like habitats of waterbirds. The present study is the first report on the cestode infection of Artemia franciscana Kellogg in Salar de Atacama lagoons in northern Chile. A total of 23 parasite larvae were isolated and identified as belonging to five cestode taxa of the order Cyclophyllidea: two species of the family Hymenolepididae, i.e. Flamingolepis sp. 1 and Flamingolepis sp. 2 (adults parasitic in flamingos); two species of Dilepididae, i.e. Fuhrmannolepis averini (adults parasitic in phalaropes) and Eurycestus avoceti (adult parasitic in charadriforms birds); and one species of Progynotaeniidae, i.e. Gynandrotaenia (?) stammeri (adult parasitic in flamingos). The cysticercoids of each species are described and figured. The study represents the first geographical record of the genera Eurycestus, Gynandrotaenia and Fuhrmannolepis in South America and the first report of Gynandrotaenia and Flamingolepis in A. franciscana in its native range. This survey also contributes to the knowledge of cestodes of Phoenicopteriformes and Charadriiformes and their life cycles in the Neotropical Region. A review of cestodes recorded in brine shrimps of the genus Artemia in the world is provided. Further studies on cestode fauna of aquatic birds and their intermediate hosts in hypersaline habitats of the Neotropical Region are needed to understand their functional role in such extreme and unique ecosystems.

First report of cestode infection in the crustacean Artemia persimilis from Southern Chilean Patagonia and its relation with the Neotropical aquatic birds.

The brine shrimp genus Artemia Leach (Crustacea, Branchiopoda), a keystone group in hipersaline wetlands all over the world, offers an excellent model to study species interactions (parasitism) and to explore "hidden fauna" (avian endoparasites). The present study is the first report on the parasite infection of the South American species Artemia persimilis from the Southern Chilean Patagonia (50°S-53°S). Samples were collected in Los Cisnes and Amarga lagoons, the two most austral populations of this crustacean described to date, during two seasons (spring and autumn). A total of 98 larvae of cestodes of the family Hymenolepididae (Cestoda, Cyclophyllidea) were found and identified as belonging to the following taxa: Confluaria podicipina (adult parasitic in grebes), Flamingolepis sp. (a cestode parasite of flamingos), Fimbriarioides (?) sp. (adults of the species of this genus infect waterfowl and shorebirds) and Wardium sp. (definitive host unknown, most probably charadriiform birds). This is a new geographical record of C. podicipina and the genus Fimbriarioides for the Neotropical Region, the latter being the most widely distributed species at both localities and seasons surveyed, and the only species recorded in autumn (April). Cestode community composition in Los Cisnes population was characterised by dominance of Flamingolepis sp., representing more than 65% of the total cestode species recorded, whereas in the Amarga population the most abundant parasite (>83%) was Fimbriarioides (?) sp. Significant seasonal variations were detected in Los Cisnes lagoon for Flamingolepis sp. and C. podicipina, with exclusive presence of them in spring (November). Besides providing novel information on cestodes infection in A. persimilis, this study provides new data on the life cycle of cestodes of Neotropical aquatic birds such as South American flamingos and grebes. Our finding expands the knowledge on the biodiversity and population dynamics of extreme and unique environments from high latitudes (Patagonia) and makes evident the need of further taxonomical and ecological studies for better understanding the life cycles of avian helminth parasites in the Neotropics and the role of aquatic invertebrates in them.

Phylogeny of hymenolepidid cestodes (Cestoda: Cyclophyllidea) from mammalian hosts based on partial 28S rDNA, with focus on parasites from shrews.

The aims of the study are to enrich the partial 28S rDNA dataset for hymenolepidids by adding new sequences for species parasitic in the genera Sorex, Neomys and Crocidura (Soricidae) and to propose a new hypothesis for the relationships among mammalian hymenolepidids. New sequences were obtained for Coronacanthus integrus, C. magnihamatus, C. omissus, C. vassilevi, Ditestolepis diaphana, Lineolepis scutigera, Spasskylepis ovaluteri, Staphylocystis tiara, S. furcata, S. uncinata, Vaucherilepis trichophorus and Neoskrjabinolepis sp. The phylogenetic analysis (based on 56 taxa) confirmed the major clades identified by Haukisalmi et al. (Zool Scr 39:631-641, 2010) based on analysis of 31 species: Ditestolepis clade, Hymenolepis clade, Rodentolepis clade and Arostrilepis clade; however, the support was weak for the early divergent lineages of the tree and for the Arostrilepis clade. Novelties revealed include the molecular evidence for the monophyly of Coronacanthus, the non-monophyletic status of Staphylocystis and the polyphyly of Staphylocystoides. The analysis has confirmed the monophyly of Hymenolepis, the monophyly of hymenolepidids from glirids, the position of Pararodentolepis and Nomadolepis as sister taxa, the polyphyly of Rodentolepis, the position of Neoskrjabinolepis and Lineolepis as sister taxa, and the close relationship among the genera with the entire reduction of rostellar apparatus. Resolved monophyletic groups are supported by the structure of the rostellar apparatus. The diversification of the Ditestolepis clade is associated with soricids. The composition of the other major clades suggests multiple evolutionary events of host switching, including between different host orders. The life cycles of Coronacanthus and Vaucherilepis are recognised as secondarily aquatic as these taxa are nested in terrestrial groups.

Hymenolepis microstoma (Cestoda: Hymenolepididae) en ratones caseros ( Mus musculus ) de Lima, Perú / Hymenolepis microstoma (Cestoda: Hymenolepididae) in house mice (Mus musculus) from Lima, Peru

Un total de 12 cestodos adultos se colectaron de los conductos biliares de ratones domésticos (Mus musculus) provenientes de Lima, Perú. Diversas características del escólex y proglotis maduros del cestodo fueron observadas para la identificación morfológica. Así mismo, se realizó un diagnóstico molecular mediante un PCR y secuenciación parcial del gen mitocondrial citocromo c oxidasa subunidad 1 (cox1). Todos los cestodos fueron identificados como Hymenolepis microstoma por morfología y métodos moleculares. El aislado de H. microstoma de Perú mostró una similitud de secuencia significativa (> 99%) con los aislados de H. microstoma previamente reportados. Nuestro informe confirma la presencia del parásito en ratones de Lima.

A total of 12 adult cestodes were collected from the bile ducts of domestic mice (Mus musculus) from Lima, Peru. Various features of the scolex and mature proglottids of the tapeworms were observed for morphological identification. A molecular diagnosis was performed by PCR-based partial sequencing of mitochondrial gene of cytochrome c oxidase subunit 1 (cox1). All cestodes were identified as Hymenolepis microstoma by morphology and molecular methods. The H. microstoma isolate from Peru showed significant sequence similarity with previously reported isolates of H. microstoma (>99%). Our report confirms the presence of the parasite in mice from Lima.

Postembryonic development of Arostrilepis microtis Gulyaev et Chechulin, 1997 (Eucestoda: Hymenolepididae) in springtails (Collembola: Entomobryidae).

The development of the metacestode stages of Arostrilepis microtis (Eucestoda: Hymenolepididae) studied on the basis of an experimental infection of springtails (Collembola: Entomobryidae) is described for the first time. A complete description of the ontogeny from oncosphere to fully a developed metacestode is given. The postembryonic development within the intermediate host includes one invagination. Cysts are without an anterior invagination pore, and the cercomer does not separate after the full maturation of larva. The comparison of morphology and maturation rate of larval cestodes of A. microtis with A. beringiensis and A. tenuicirrosa is presented. Cysticercoids of A. microtis are significantly larger and require a longer period of development than those of A. beringiensis and A. tenuicirrosa.

Characterization of the complete mitochondrial genome of the cloacal tapeworm Cloacotaenia megalops (Cestoda: Hymenolepididae).

BACKGROUND: The cloacal tapeworm Cloacotaenia megalops (Hymenolepididae) is one of the most common cestode parasites of domestic and wild ducks worldwide. However, limited information is available regarding its epidemiology, biology, genetics and systematics. This study provides characterisation of the complete mitochondrial (mt) genome of C. megalops. METHODS: The complete mt genome of C. megalops was obtained by long PCR, sequenced and annotated. RESULTS: The length of the entire mt genome of C. megalops is 13,887 bp; it contains 12 protein-coding, 2 ribosomal RNA and 22 transfer RNA genes, but lacks an atp8 gene. The mt gene arrangement of C. megalops is identical to that observed in Anoplocephala magna and A. perfoliata (Anoplocephalidae), Dipylidium caninum (Dipylidiidae) and Hymenolepis diminuta (Hymenolepididae), but differs from that reported in taeniids owing to the position shift between the tRNA (L1) and tRNA (S2) genes. The phylogenetic position of C. megalops was inferred using Maximum likelihood and Bayesian inference methods based on the concatenated amino acid data for 12 protein-coding genes. Phylogenetic trees showed that C. megalops is sister to Anoplocephala spp. (Anoplocephalidae) + Pseudanoplocephala crawfordi + Hymenolepis spp. (Hymenolepididae) indicating that the family Hymenolepididae is paraphyletic. CONCLUSIONS: The complete mt genome of C. megalops is sequenced. Phylogenetic analyses provided an insight into the phylogenetic relationships among the families Anoplocephalidae, Hymenolepididae, Dipylidiidae and Taeniidae. This novel genomic information also provides the opportunity to develop useful genetic markers for studying the molecular epidemiology, biology, genetics and systematics of C. megalops.

Description and phylogenetic affinities of two new species of Nomadolepis (Eucestoda, Hymenolepididae) from Eastern Palearctic.

Two new species of hymenolepidid cestodes belonging to the genus Nomadolepis are described from small mammals in western Siberia and the Far East, Russian Federation. Nomadolepis fareasta n. sp. is described from the striped dwarf hamster Cricetulus barabensis in Amurskaya Oblast' and Nomadolepis shiloi n. sp. is described from the Eurasian harvest mouse Micromys minutus from Novosibirskaya Oblast' and Amurskaya Oblast'. Nomadolepis fareasta n. sp. differs from Nomadolepis merionis, Nomadolepis ellobii and N. shiloi n. sp. in having a substantially longer strobila, longer cirrus-sac and wider ovary. Furthermore, N. fareasta n. sp. can be readily distinguished from its congeners by the arrangement of the testes and the number and size of rostellar hooks. Nomadolepis shiloi n. sp. differs from N. merionis, N. ellobii and N. fareasta n. sp. by the number and length of the rostellar hooks, the presence of irregular transverse anastomoses as well as the length of the cirrus-sac and position of the cirrus-sac in relation to the poral ventral osmoregulatory canal. Morphological differentiation of the two new species from morphologically similar Palearctic species of the related genus Rodentolepis (sensu lato) from rodents is also provided. Phylogenetic affinities of Nomadolepis were studied for the first time using partial sequences of the nuclear ribosomal 28S DNA gene. Phylogenetic analysis strongly supported the status of Nomadolepis as a separate genus closest to Pararodentolepis.