First record of Metapolystoma (Monogenea: Polystomatidae) from Boophis tree frogs in Madagascar, with the description of five new species.

Although Madagascar has more than 350 frog species of which all but two are endemic to the island, the known polystome (Monogenea: Polystomatidae) diversity parasitizing Malagasy frogs is low, encompassing five species of Madapolystoma, one species of Kankana and one Metapolystoma. Investigating the parasite diversity of frog parasites at selected Malagasy localities led to the discovery of undescribed polystomes. Five treefrogs, Boophis albilabris, Boophis doulioti, Boophis luteus, Boophis madagascariensis and Boophis occidentalis were found to be infected and are reported here as hosts for new Metapolystoma species. Morphological investigation, combining examination of body length, haptor length, genital bulb width, genital crown diameter, genital spine number, genital spine length, ovary length, egg length, hamulus length, hamulus guard length and hamulus hook length, revealed five distinct morphotypes. Phylogenetic analysis and genetic divergences obtained for three of the five morphotypes, support the distinction of new species. Metapolystoma ansuanum n. sp. is described from B. luteus, Metapolystoma falcatum n. sp. from B. doulioti, Metapolystoma multiova n. sp. from B. occidentalis, Metapolystoma theroni n. sp. from B. madagascareniensis and Metapolystoma vencesi n. sp. from B. albilabris. Finally, although the validity of Metapolystoma as taxon is not fully resolved yet, the phylogenetic position of the described species and their morphology provide clear evidence for new metapolystome taxa.

Widespread presence of the pathogenic fungus Batrachochytrium dendrobatidis in wild amphibian communities in Madagascar.

Amphibian chytridiomycosis, an emerging infectious disease caused by the fungus Batrachochytrium dendrobatidis (Bd), has been a significant driver of amphibian declines. While globally widespread, Bd had not yet been reported from within Madagascar. We document surveys conducted across the country between 2005 and 2014, showing Bd's first record in 2010. Subsequently, Bd was detected in multiple areas, with prevalence reaching up to 100%. Detection of Bd appears to be associated with mid to high elevation sites and to have a seasonal pattern, with greater detectability during the dry season. Lineage-based PCR was performed on a subset of samples. While some did not amplify with any lineage probe, when a positive signal was observed, samples were most similar to the Global Panzootic Lineage (BdGPL). These results may suggest that Bd arrived recently, but do not exclude the existence of a previously undetected endemic Bd genotype. Representatives of all native anuran families have tested Bd-positive, and exposure trials confirm infection by Bd is possible. Bd's presence could pose significant threats to Madagascar's unique "megadiverse" amphibians.

First monogenean flatworm from a microhylid frog host: Kankana, a new polystome genus from Madagascar.

Kankana manampoka n. gen., n. sp. (Monogenea, Polystomatidae), is described from the urinary bladder of the narrow-mouthed frog Platypelis pollicaris. This is the first record of a polystome from the Microhylidae and the third polystome genus from Madagascar, next to Metapolystoma and Madapolystoma. The extensive uterus and presence of hamuli resemble Metapolystoma but the vitellarium confined to the lateral fields in Kankana is different. Madapolystoma also has an extensive uterus but contain only up to 32 advanced developed larvae. Based on the extensive uterus filling the body proper and the vitellarium confined to two lateral fields posterior in the body this new polystome resembles Eupolystoma known from Africa and India. However, unlike Eupolystoma, the gonads are in the middle of the body, vaginae are lacking and a pair of hamuli is present. A molecular phylogenetic analysis of concatenated 18S and 28S ribosomal RNA gene sequences supplemented by genetic distances inferred from 28S and COI sequences showed that this new genus is more related to Madapolystoma, a genus only reported from Madagascar, than to Eupolystoma known from Africa and India and Metapolystoma known from Africa and Madagascar.

The double odyssey of Madagascan polystome flatworms leads to new insights on the origins of their amphibian hosts.

Polystomatid flatworms are parasites of high host specificity, which mainly infect amphibian hosts. Only one polystome species has so far been recorded from Madagascar despite the high species richness and endemicity of amphibians on this island. Out of the 86 screened Malagasy frog species, we recovered polystomes from 25 in the families Ptychadenidae and Mantellidae. Molecular phylogenetic analysis uncovered an unexpected diversity of polystome species belonging to two separate clades: one forming a lineage within the genus Metapolystoma, with one species in Ptychadena and several species in the mantellid host genera Aglyptodactylus and Boophis; and the second corresponding to an undescribed genus that was found in the species of the subfamily Mantellinae in the family Mantellidae. The phylogenetic position of the undescribed genus along with molecular dating suggests that it may have colonized Madagascar in the Late Mesozoic or Early Cainozoic. By contrast, the more recent origin of Metapolystoma in Madagascar at ca 14-2 Myr ago strongly suggests that the ancestors of Ptychadena mascareniensis colonized Madagascar naturally by overseas dispersal, carrying their Metapolystoma parasites. Our findings provide a striking example of how parasite data can supply novel insights into the biogeographic history of their hosts.

High mitochondrial diversity within and among populations of Malagasy poison frogs.

The diurnal, brightly colored, and toxic frogs of the genus Mantella are among the most prominent representatives of the endemic anuran fauna of Madagascar. Especially three closely related species, M. aurantiaca, M. crocea, and M. milotympanum, are intensively collected for the pet trade, although basic data on their natural history and genetic diversity are still lacking. Our phylogenetic analyses based on 2.8 kbp of partial 16S rRNA, 12S rRNA, cytochrome b, and rhodopsin DNA sequences confirmed that these species belong to one of the five major clades in Mantella, the M. madagascariensis group. A haplotype network constructed using 830 bp of cytochrome b in 49 individuals from seven populations revealed that M. milotympanum and M. crocea have largely similar haplotypes sharing, confirming doubts about the species validity of M. milotympanum and indicating independent evolution of bright orange pattern in M. milotympanum and M. aurantiaca. Further, clustering of four individuals of M. aurantiaca from Andranomena with M. crocea suggests incomplete lineage sorting or introgression resulting from secondary contact of refugial populations. AMOVA confirmed significant intrapopulation nucleotide diversity (>20%). These diversity patterns and our field observations indicate relatively large population sizes. Hence, overcollecting is probably a minor problem and conservation efforts should rather focus on saving some large populations from habitat destruction through logging and forest fires.