BackgroundNeotropical freshwater stingrays (Batoidea: Potamotrygonidae) host a diverse parasite fauna, including cestodes. Both cestodes and their stingray hosts are marine-derived, but the taxonomy of this host/parasite system is poorly understood.MethodologyMorphological and molecular (Cytochrome oxidase I) data were used to investigate diversity in freshwater lineages of the cestode genus Rhinebothrium Linton, 1890. Results were based on a phylogenetic hypothesis for 74 COI sequences and morphological analysis of over 400 specimens. Cestodes studied were obtained from 888 individual potamotrygonids, representing 14 recognized and 18 potentially undescribed species from most river systems of South America.ResultsMorphological species boundaries were based mainly on microthrix characters observed with scanning electron microscopy, and were supported by COI data. Four species were recognized, including two redescribed (Rhinebothrium copianullum and R. paratrygoni), and two newly described (R. brooksi n. sp. and R. fulbrighti n. sp.). Rhinebothrium paranaensis Menoret & Ivanov, 2009 is considered a junior synonym of R. paratrygoni because the morphological features of the two species overlap substantially. The diagnosis of Rhinebothrium Linton, 1890 is emended to accommodate the presence of marginal longitudinal septa observed in R. copianullum and R. brooksi n. sp. Patterns of host specificity and distribution ranged from use of few host species in few river basins, to use of as many as eight host species in multiple river basins.SignificanceThe level of intra-specific morphological variation observed in features such as total length and number of proglottids is unparalleled among other elasmobranch cestodes. This is attributed to the large representation of host and biogeographical samples. It is unclear whether the intra-specific morphological variation observed is unique to this freshwater system. Nonetheless, caution is urged when using morphological discontinuities to delimit elasmobranch cestode species because the amount of variation encountered is highly dependent on sample size and/or biogeographical representation.

Helminths of the spiral intestine of neotropical freshwater stingrays (Potamotrygonidae) were examined in Peru for the first time. The stingrays examined for helminths included Paratrygon aiereba, Potamotrygon motoro, and Potamotrygon cf. castexi. Present in P. aiereba were the cestodes Nandocestus guariticus (Marques, Brooks, and Lasso, 2001) n. gen. n. comb., Rhinebothrium copianullum n. sp., Rhinebothrium sp. 1, Rhinebothroides sp., Potamotrygonocestus cf. fitzgeraldae, and 1 species each of Cucullanus and Rhabdochona. Nandocestus n. gen. is erected to house N. guariticus, which is formally transferred from Anindobothrium Marques, Brooks, and Lasso, 2001. The new genus is unique among phyllobothriids in its possession of circummedullary vitelline follicles and a submarginal genital pore, in combination with bothridia with a single apical sucker and marginal bothridial loculi. The helminths of P. motoro included the cestodes Paraoncomegas araya, Acanthobothrium peruviense n. sp., Acanthobothrium cf. ramiroi, Rhinebothrium sp. 1, Rhinebothroides sp. 1, Potamotrygonocestus sp., the nematode Brevimulticaecum regoi, a species of Cucullanus, and a species of the digenean superfamily Diplostomoidea. The helminths of P. cf. castexi included the cestodes P. araya, N. guariticus n. gen. n. comb., Acanthobothrium cf. peruviense, Potamotrygonocestus sp., Rhinebothrium sp. 1, Rhinebothroides sp. 2, the nematode species Echinocephalus daileyi and B. regoi, 1 species each of Cucullanus, Rhabdochona, and Procamallanus, and a species of the digenean superfamily Hemiuroidea. All taxa were examined via light microscopy; the cestode taxa were also examined using scanning electron microscopy. Each helminth species recorded in this study is a first report from Peru. The study suggests that the diversity and host specificity of the cestodes in potamotrygonid stingrays may be greater than previously thought. The known numbers of genera and species of tetraphyllidean cestodes parasitizing neotropical freshwater stingrays are now 6 and 22, respectively.

Adult tetraphyllidean tapeworms (Platyhelminthes: Eucestoda) from the spiral intestines of 3 species of potamotrygonid stingrays (Paratrygon aiereba, Potamotrygon castexi, and Portamotrygon motoro) in the Madre de Dios river in Peru were found to host numerous cysts embedded in their parenchymal tissues. Histological sections of the cysts revealed the presence of a scolex bearing 4 suckers and an unarmed apical organ consistent with larval stages of both Cyclophyllidea and Proteocephalidea. To further elucidate their identities, partial 28S ribosomal DNA (rDNA) sequences were characterized from 3 cysts and 4 adult Rhinebothrium spp. 'host' worms and screened against all available cestode 28S rDNA data. Initial BLAST screening and subsequent alignment ruled out the possibility that the cysts were cyclophyllidean, and the cyst and adult sequences were thus aligned together with all available lecanicephalidean, litobothriidean, proteocephalidean, and tetraphyllidean sequences. Sequences from all 3 cysts were identical, and phylogenetic analysis clearly placed them among derived members of the Proteocephalidea, although no exact match was found. Sequences from the adult host worms formed 2 identical pairs and grouped together with other tetraphyllidean species from rays. These results are compared with records of hyperparasites of South American catfish cestodes. This is the first confirmed record of a proteocephalidean cestode parasitizing a tetraphyllidean cestode.

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