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Background: Polystomatid flatworms in chelonians are divided into three genera, i.e. Polystomoides Ward, 1917, Polystomoidella Price, 1939 and Neopolystoma Price, 1939, according to the number of haptoral hooks. Among the about 55 polystome species that are known to date from the 327 modern living chelonians, only four species of Polystomoides are currently recognised within the 45 South American freshwater turtles., Methods: During 2012, several sites in the vicinity of the cities Cayenne and Kaw in French Guiana were investigated for freshwater turtles. Turtles were collected at six sites and the presence of polystomatid flatworms was assessed from the presence of polystome eggs released by infected specimens., Results: Among the three turtle species that were collected, no polystomes were found in the gibba turtle Mesoclemmys gibba (Schweigger, 1812). The spot-legged turtle Rhinoclemmys punctularia (Daudin, 1801) was infected with two species of Neopolystoma Price, 1939, one in the conjunctival sacs and the other in the urinary bladder, while the scorpion mud turtle Kinosternon scorpioides (Linnaeus, 1766) was found to be infected with a single Neopolystoma species in the conjunctival sacs. These parasites could be distinguished from known species of Neopolystoma by a combination of morphological characteristics including body size, number and length of genital spines, shape and size of the testis. They were also differentiated at the molecular level using the cox1 gene marker. Based on morphological and genetic evidences, three new species are described herein, namely Neopolystoma cayensis n. sp. and Neopolystoma guianensis n. sp. from the bladder and the conjunctival sacs of R. punctularia, respectively, and Neopolystoma scorpioides n. sp. from the conjunctival sacs of K. scorpioides. However the monophyly of Polystomoides and Neopolystoma is still questioned regarding their phylogeny based on a dataset comprising four concatenated genes, namely, 18S, 28S and 12S rRNA genes and cox1., Conclusions: In addition to these being the first chelonian polystomes to be reported and described from French Guiana, they represent the first polystomes from the hosts K. scorpioides and R. punctularia and the first representatives of Neopolystoma from South America. Chelonian polystomes now require an in-depth morphological study to reconcile the taxonomy of the genera with species evolution.

Background: The phylum Platyhelminthes (flatworms) contains an important group of bilaterian organisms responsible for many debilitating and chronic infectious diseases of human and animal populations inhabiting the planet today. In addition to their biomedical and veterinary relevance, some platyhelminths are also frequently used models for understanding tissue regeneration and stem cell biology. Therefore, the molecular (genetic and epigenetic) characteristics that underlie trophic specialism, pathogenicity or developmental maturation are likely to be pivotal in our continued studies of this important metazoan group. Indeed, in contrast to earlier studies that failed to detect evidence of cytosine or adenine methylation in parasitic flatworm taxa, our laboratory has recently defined a critical role for cytosine methylation in Schistosoma mansoni oviposition, egg maturation and ovarian development. Thus, in order to identify whether this epigenetic modification features in other platyhelminth species or is a novelty of S. mansoni, we conducted a study simultaneously surveying for DNA methylation machinery components and DNA methylation marks throughout the phylum using both parasitic and non-parasitic representatives., Results: Firstly, using both S. mansoni DNA methyltransferase 2 (SmDNMT2) and methyl-CpG binding domain protein (SmMBD) as query sequences, we illustrate that essential DNA methylation machinery components are well conserved throughout the phylum. Secondly, using both molecular (methylation specific amplification polymorphism, MSAP) and immunological (enzyme-linked immunoabsorbent assay, ELISA) methodologies, we demonstrate that representative species (Echinococcus multilocularis, Protopolystoma xenopodis, Schistosoma haematobium, Schistosoma japonicum, Fasciola hepatica and Polycelis nigra) within all four platyhelminth classes (Cestoda, Monogenea, Trematoda and 'Turbellaria') contain methylated cytosines within their genome compartments., Conclusions: Collectively, these findings provide the first direct evidence for a functionally conserved and enzymatically active DNA methylation system throughout the Platyhelminthes. Defining how this epigenetic feature shapes phenotypic diversity and development within the phylum represents an exciting new area of metazoan biology.