Codon usage in the flatworm Schistosoma mansoni is shaped by the mutational bias towards A+T and translational selection, which increases GC-ending codons in highly expressed genes.

Schistosoma mansoni is a trematode flatworm that parasitizes humans and produces a disease called bilharzia. At the genomic level, it is characterized by a low genomic GC content and an "isochore-like" structure, where GC-richest regions, mainly placed at the extremes of the chromosomes, are interspersed with low GC-regions. Furthermore, the GC-richest regions are at the same time the gene-richest, and where the most heavily expressed genes are placed. Taking these features into account, we decided to reanalyze the codon usage of this flatworm. Our results show that a) when all genes are considered together, the strong mutational bias towards A + T leads to a predominance of A/T-ending codons, b) a multivariate analysis discriminates between highly and lowly expressed genes, c) the sequences expressed at highest levels display a significant increase in G/C-ending codons, d) when comparing the molecular distances with a closely related species the synonymous distance in highly expressed genes is significantly lower than in lowly expressed sequences. Therefore, we conclude that despite previous results, which were performed with a small sample of genes, codon usage in S. mansoni is the result of two forces that operate in opposite directions: while mutational bias leads to a predominance of A/T codons, translational selection, working at the level of speed, increment G/C ending triplets.

Lost and Found: Piwi and Argonaute Pathways in Flatworms.

Platyhelminthes comprise one of the major phyla of invertebrate animals, inhabiting a wide range of ecosystems, and one of the most successful in adapting to parasitic life. Small non-coding RNAs have been implicated in regulating complex developmental transitions in model parasitic species. Notably, parasitic flatworms have lost Piwi RNA pathways but gained a novel Argonaute gene. Herein, we analyzed, contrasted and compared the conservation of small RNA pathways among several free-living species (a paraphyletic group traditionally known as 'turbellarians') and parasitic species (organized in the monophyletic clade Neodermata) to disentangle possible adaptations during the transition to parasitism. Our findings showed that complete miRNA and RNAi pathways are present in all analyzed free-living flatworms. Remarkably, whilst all 'turbellarians' have Piwi proteins, these were lost in parasitic Neodermantans. Moreover, two clusters of Piwi class Argonaute genes are present in all 'turbellarians'. Interestingly, we identified a divergent Piwi class Argonaute in free living flatworms exclusively, which we named 'Fliwi'. In addition, other key proteins of the Piwi pathways were conserved in 'turbellarians', while none of them were detected in Neodermatans. Besides Piwi and the canonical Argonaute proteins, a flatworm-specific class of Argonautes (FL-Ago) was identified in the analyzed species confirming its ancestrallity to all Platyhelminthes. Remarkably, this clade was expanded in parasitic Neodermatans, but not in free-living species. These phyla-specific Argonautes showed lower sequence conservation compared to other Argonaute proteins, suggesting that they might have been subjected to high evolutionary rates. However, key residues involved in the interaction with the small RNA and mRNA cleavage in the canonical Argonautes were more conserved in the FL-Agos than in the Piwi Argonautes. Whether this is related to specialized functions and adaptations to parasitism in Neodermatans remains unclear. In conclusion, differences detected in gene conservation, sequence and structure of the Argonaute family suggest tentative biological and evolutionary diversifications that are unique to Platyhelminthes. The remarkable divergencies in the small RNA pathways between free-living and parasitic flatworms indicate that they may have been involved in the adaptation to parasitism of Neodermatans.

First study of the Polycladida (Rhabditophora, Platyhelminthes) from the Pacific Coast of Costa Rica.

In the present work was carried out in the intertidal zone of Las Baulas de Guanacaste National Marine Park (PNMB) located on the Pacific coast of Costa Rica.                The main objective was to contribute to knowledge about the invertebrate diversity of the park, one of the richest bioregions on the planet, about which little is known. This study assesses the Order Polycladida Lang, 1884, a cornerstone of this ecosystem and one of the most cosmopolitan and plastic invertebrate taxa in the animal kingdom.                In total, 57 individuals were collected in the rocky intertidal zone of Carbón and Langosta beaches. Nine different species were identified, of which four are new for Costa Rica: Semonia bauliensis n. sp.; Cryptostylochus sesei n. sp.; Paraplanocera angeli n. sp., Prostheceraeus fitae n. sp.; and five new records: Paraplanocera oligoglena (Schmarda, 1859); Marcusia ernesti Hyman, 1953; Enchiridium magec Cuadrado, Moro Noreña, 2017; Pseudobiceros bajae (Hyman, 1953); and the genus Boninia spp.

Revision and description of six species of Choeradoplana (Platyhelminthes, Tricladida), with an emendation to the genus.

Living representatives of the Neotropical genus Choeradoplana Graff, 1896 (Geoplaninae, Tricladida, Platyhelminthes) are easily recognized by the typical shape of the head which is laterally expanded, rolled-up, and ventrally provided with two glandular cushions. In this study, the morphology and phylogeny (cytochrome C oxidase subunit I gene) of several species of land planarians are taxonomically investigated. Four of the six species studied are new to science, namely: Ch. eudoxiae Silva & Carbayo, sp. nov., Ch. claudioi Lago-Barcia & Carbayo, sp. nov., Ch. onae Lago-Barcia & Carbayo, sp. nov., and Ch. riutortae Lago-Barcia & Carbayo, sp. nov. The species Choeradoplana albonigra and Ch. eudoxiae deviate from the usual body shape pattern in that the head does not present lateral expansions nor glandular cushions, becoming indistinguishable from its sister genus Cephaloflexa. Pseudogeoplana tristriata (Schultze & Müller, 1857) is also redescribed from a newly collected specimen and was discovered to be a member of Choeradoplana. Graff (1899) also studied another specimen that was considered to be conspecific with P. tristriata; however, in this new it is concluded that it is not conspecific but rather a new species. The name Pseudogeoplana aevipandemiae Lago-Barcia & Carbayo, sp. nov. is suggested for Graff's specimen.

Stage-specific transcriptomic analysis of the model cestode Hymenolepis microstoma.

Most parasitic flatworms go through different life stages with important physiological and morphological changes. In this work, we used a transcriptomic approach to analyze the main life-stages of the model tapeworm Hymenolepis microstoma (eggs, cysticercoids, and adults). Our results showed massive transcriptomic changes in this life cycle, including key gene families that contribute substantially to the expression load in each stage. In particular, different members of the cestode-specific hydrophobic ligand-binding protein (HLBP) family are among the most highly expressed genes in each life stage. We also found the transcriptomic signature of major metabolic changes during the transition from cysticercoids to adult worms. Thus, this work contributes to uncovering the gene expression changes that accompany the development of this important cestode model species, and to the best of our knowledge represents the first transcriptomic study with robust replicates spanning all of the main life stages of a tapeworm.

Cataloguing Prof. Eudóxia M. Froehlich's wet collection of land planarians (Platyhelminthes, Geoplanidae): a bounty of type specimens of 78 species.

Zoologist Prof. Eudóxia Maria Froehlich (1929*-2015†) kept an uncatalogued personal collection of land planarians in wet medium. We catalogued their contents. Glass jars contained a variable number of small vials, each with one or several labeled specimens. We transcribed the species names and field data, when available, written on card labels to a table. A total of 1772 vials in 80 jars, contained over 2700 specimens. Subsequently, we cross checked field data with that of the type specimens. As a result, we identified, with different degrees of certainty, type specimens of 78 species. These types compose a very significant potential source of additional morphological details of the species to conform with current standards in the taxonomy of the group.

Five new pseudocryptic land planarian species of Cratera (Platyhelminthes: Tricladida) unveiled through integrative taxonomy.

BACKGROUND: Cratera is a genus of land planarians endemic to the Brazilian Atlantic forest. The species of this genus are distinguished from each other by a series of external and internal characters, nonetheless they represent a challenging taxonomic issue due to the extreme alikeness of the species analysed in the present work. To resolve these difficulties, we have performed morphological analyses and used three nuclear markers (ribosomal 18S and 28S, Elongation Factor, a new anonymous marker named Tnuc813) and two mitochondrial fragments (Cytochrome c oxidase subunit I gene, and a fragment encompasing NADH deshydrogenase subunit 4 gene, trnF and the beginning of the Cytochrome c oxidase I gene) in an integrative taxonomic study. METHODS: To unveil cryptic species, we applied a molecular species delimitation approach based on molecular discovery methods, followed by a validation method. The putative species so delimited were then validated on the basis of diagnostic morphological features. RESULTS: We discovered and described four new species, namely Cratera assu, C. tui, C. boja, and C. imbiri. A fifth new species, C. paraitinga was not highly supported by molecular evidence, but was described because its morphological attributes are unique. Our study documents for the genus Cratera the presence of a number of highly similar species, a situation that is present also in other genera of land planarians. The high number of poorly differentiated and presumably recent speciation events might be explained by the recent geological history of the area.

Trying to take over the world: Potential distribution of Obama nungara (Platyhelminthes: Geoplanidae), the Neotropical land planarian that has reached Europe.

Obama nungara Carbayo, Álvarez-Presas, Jones, & Riutort, 2016 is a land planarian (Platyhelminthes: Geoplanidae) native to southern South America, which has recently dispersed towards several countries of the European continent, thus becoming a threat to the native soil fauna. Its dispersion would be favoured by its wide food habit and its tendency to live linked to humans, being the plant trade its most plausible vector of dispersion. Here, we explored the potential distribution of O. nungara on a global scale by using the MaxEnt software. We used 144 records (encompassing 10 countries) from sampling campaigns, citizen science, recent literature, and material deposited in scientific collections. Our results showed that southern South America has favourable climatic conditions for O. nungara. MaxEnt also allowed predicting expansions to countries of Europe where this planarian is already established and to others not yet colonized, as well as to Asia (southern coast of the Caspian Sea, Taiwan, and south-east of mainland China) and Oceania (south-east of Australia and New Zealand). The potential distribution of O. nungara was mainly outlined by climatic factors related to temperature (annual mean temperature, mean temperature of the coldest quarter, and annual temperature range). Thus, under a global warming scenario, a significant expansion of O. nungara relative to the current prediction is expected. This information may be useful to design strategies to prevent new introductions, since the dissemination of this planarian seems to be strongly man-linked.

Genome-wide identification of ABC transporters in monogeneans.

ATP-Binding Cassette (ABC) transporters are proteins that actively mediate the transport of a wide variety of molecules, including drugs. Thus, in parasitology, ABC transporters have gained attention as potential targets for therapeutic drugs. Among the parasitic Platyhelminthes, ABC transporters have been identified and classified in a few species of Trematoda and Cestoda but not in Monogenea. Monogeneans are mainly ectoparasites of marine and freshwater fish, although they can also be found on other aquatic organisms. Severe epizootics caused by monogeneans have been reported around the world, mainly in confined and/or overcrowded fish. The purpose of this study was to identify the ABC transporters in four species of monogeneans (Gyrodactylus salaris, Protopolystoma xenopodis, Eudiplozoon nipponicum and Neobenedenia melleni) for which genomic resources are publicly available. For comparative purposes, ABC transporters were also identified in endoparasitic (Schistosoma mansoni and Echinococcus granulosus) and free-living (Macrostomun lignano and Schmidtea mediterranea) platyhelminths. Thirty-two putative ABC transporters were identified in the genome of G. salaris, 40 in the genome of P. xenopodis, 46 in the transcriptome of E. nipponicum and 9 in a rather limited ESTs set available for N. melleni. Of the eight ABC subfamilies (A-H) known in metazoans, subfamily H was the only one not found in any monogenean species. In contrast, ABCC was the best represented subfamily. Phylogenetic analyses showed a few cases of one-to-one orthologous relationships, which agree with results from other metazoan species. We found some monogenean ABC members related to subfamilies B, C and G involved in drug resistance in humans. This information may be useful for future functional studies on ABC transporters in monogeneans.

Two new land planarian species (Platyhelminthes: Tricladida: Geoplanidae) from protected areas in southern Brazil.

We describe two new species of Neotropical land planarians found in protected areas of the Atlantic Forest in southern Brazil. Cratera obsidiana Amaral, Boll Leal-Zanchet, sp. nov., found in the State Park of Turvo, is the first species of the genus Cratera described from an area of deciduous seasonal forest. It can be distinguished from its congeners by the uniformly black dorsal color and the prostatic vesicle with proximal portion laterally expanded and directed upward. Luteostriata subtilis Boll, Amaral Leal-Zanchet, sp. nov., found in the National Park of Aparados da Serra, in an area covered by Araucaria moist forest, is distinguished from its congeners by only two conspicuous longitudinal stripes on the dorsal surface of the body and a large fold in the male atrium below the opening of the ejaculatory duct. We also provide an updated key for species of Cratera and the first key for species of Luteostriata.

Compositional Analysis of Flatworm Genomes Shows Strong Codon Usage Biases Across All Classes.

In the present work, we performed a comparative genome-wide analysis of 22 species representative of the main clades and lifestyles of the phylum Platyhelminthes. We selected a set of 700 orthologous genes conserved in all species, measuring changes in GC content, codon, and amino acid usage in orthologous positions. Values of 3rd codon position GC spanned over a wide range, allowing to discriminate two distinctive clusters within freshwater turbellarians, Cestodes and Trematodes respectively. Furthermore, a hierarchical clustering of codon usage data differs remarkably from the phylogenetic tree. Additionally, we detected a synonymous codon usage bias that was more dramatic in extreme GC-poor or GC-rich genomes, i.e., GC-poor Schistosomes preferred to use AT-rich terminated synonymous codons, while GC-rich M. lignano showed the opposite behavior. Interestingly, these biases impacted the amino acidic usage, with preferred amino acids encoded by codons following the GC content trend. These are associated with non-synonymous substitutions at orthologous positions. The detailed analysis of the synonymous and non-synonymous changes provides evidence for a two-hit mechanism where both mutation and selection forces drive the diverse coding strategies of flatworms.

Molecular and morphological characterization of the metacercariae of two species of diplostomid trematodes (Platyhelminthes, Digenea) in freshwater fishes of the Batalha River, Brazil.

The Diplostomidae include a large group of flatworms with complex life cycles and are frequently found parasitizing the eyes and central nervous system of freshwater fishes. The morphological identification of the metacercariae at species level is not always possible. Thus, molecular tools have become essential to assist in the parasite species determination. This study was aimed at describing two diplostomid metacercariae found in freshwater fish in São Paulo, Brazil, based on morphological characters and in the genetic characterization of COI sequences. Our results showed that the two recognized taxa (Tylodelphys sp. and Diplostomidae gen. sp.) appear to be different from the species already described in South America. Tylodelphys sp. differs morphologically from Tylodelphys xenopi, T. mashonense, T. jenynsiae, and T. scheuringi. The metacercariae of T. clavata and T. conifera are smaller than Tylodelphys sp., while T. podicipina is larger than the metacercariae described here. The phylogenetic analysis of COI sequences yielded Tylodelphys sp. as the sister species of Tylodelphys sp. 4, a species reported from the brain of the eleotrid Gobiomorus maculatus in Oaxaca, Mexico. The metacercariae identified as Diplostomidae gen. sp. are morphologically different from the known diplostomid metacercariae and did not match with other diplostomid sequences available. Diplostomidae gen. sp. is recovered as the sister species of Diplostomum ardeae. Although the morphological evidence and the COI sequences differentiate the metacercariae found, the absence of adult specimens of both species precludes the specific designation. This is one of the first papers that use an integrative taxonomy approach to describe the species diversity of diplostomid trematodes in Brazil.

First record of Polycystididae (Platyhelminthes, Kalyptorhynchia) from Cuba, with the description of a new genus and five new species, and remarks and the description of one new species from Panama.

The first records of Polycystididae from Cuba are provided and discussed. In total nine species have been collected, five of which are new to science, one representing a new genus. Polycydora intermedia gen. n. sp. n. shows intermediate features between Polycystis Kölliker, 1845 and Paulodora Marcus, 1948. The ovaries are kidney shaped, with the oocytes arranged in a row, and lack the hard "nozzles" of Paulodora. A female bursa as in Polycystis is present. In P. intermedia gen. n. sp. n. lacks the strong muscle bulb at the male bursa and the accessory glands type I in the male atrium, which are present in Polycystis. The male atrial organs include a prostate stylet type II connected to a free prostate vesicle type II. The four new species (Phonorhynchoides minor sp. n., Phonorhynchopsis capillaris sp. n., Phonorhynchopsis sublinguatus sp. n., Myobulla armenterosi sp. n.) are distinguished from their congeners by the specific shape and length of the male hard copulatory structures. Phonorhynchoides minor sp. n. has the smallest stylets within the genus, and these stylets are more straight than those of the other species of the genus. In Phonorhynchopsis capillaris sp. n. the prostate stylet type IV is only 20% of the length of the accessory stylet type IV, the lowest relative length within the genus. The accessory stylet in this species is extremely thin; it is only 2 µm wide. Phonorhynchopsis sublinguatus sp. n. has a prostate stylet widened in the distal half, ending in a rounded tip, not twisted in the middle. Because of its strong resemblance to M. armenterosi sp. n., M. berti sp. n., a new species of Myobulla Artois Schockaert, 2000 collected at the Atlantic coast of Panama is described here too. Both species of Myobulla have a prostate stylet type III that shows a 90° turn at some point, as is the case in some other species of Myobulla. In M. berti sp. n. the stylet is smaller overall and is bent in the middle; in M. armenterosi sp. n. the stylet is larger and it is bent more distally. All of the four known species have a very wide geographical distribution: Phonorhynchopsis haegheni, Alcha evelinae, Paraustrorhynchus elixus, and Gyratrix hermaphroditus.

Taxonomic revision of four species of the Neotropical land planarian genera Cratera and Geoplana (Platyhelminthes, Tricladida) with a description of two new species and an emendation of Cratera.

The Brazilian land planarians Cratera crioula, C. joia, Geoplana hina, and G. taxiarcha (Platyhelminthes, Tricladida, Geoplanidae) are revised taxonomically from type material and additional specimens. Geoplana hina sensu Carbayo et al. (2013) was found to be an undescribed species and therefore is described and named as Cratera picuia sp. n. A new species of the genus is also described and named as Cratera arucuia sp. n. G. hina and G. taxiarcha are transferred to Cratera. The most remarkable morphological feature of Cratera-a dilated terminal portion of the ejaculatory duct-is either absent, inconspicuous, or variable in C. hina, C. joia, C. picuia sp. n., and C. arucuia sp. n. Based on the monophyletic status of Cratera inferred elsewhere, an emendation of the genus is here proposed to encompass the morphological variation observed in the genus.

Integrative approach reveals two new species of Obama (Platyhelminthes: Tricladida) from the South-Brazilian Atlantic Forest.

The genus Obama Carbayo et al., 2013 includes 38 species, being the most species-rich within Geoplaninae. Species of this genus show a similar anatomy regarding their copulatory apparatus, which may hinder species differentiation. In this study, we describe two new species, presenting a marbled colour pattern, found in two different phytophysionomies of the Atlantic Forest, namely Semi-deciduous Forest and Araucaria Forest. Both species can be distinguished from their congeners, as well from each other, by colour pattern and eye arrangement combined with characteristics of the pharynx, penis papilla and prostatic vesicle, confirmed by molecular analyses from cytochrome c oxidase subunit I gene (COI). Phylogenetic analyses suggest that both species herein studied are closely related to another species that occurs in areas of Araucaria Forest (O. maculipunctata). Results also indicate the need to use at least 600 bp of the gene COI in the definitions of interspecific divergences and for species delineation, at least for the genus Obama.

Evolutionary fates of universal stress protein paralogs in Platyhelminthes.

BACKGROUND: Universal stress proteins (USPs) are present in all domains of life. Their expression is upregulated in response to a large variety of stress conditions. The functional diversity found in this protein family, paired with the sequence degeneration of the characteristic ATP-binding motif, suggests a complex evolutionary pattern for the paralogous USP-encoding genes. In this work, we investigated the origin, genomic organization, expression patterns and evolutionary history of the USP gene family in species of the phylum Platyhelminthes. RESULTS: Our data showed a cluster organization, a lineage-specific distribution, and the presence of several pseudogenes among the USP gene copies identified. The absence of a well conserved -CCAATCA- motif in the promoter region was positively correlated with low or null levels of gene expression, and with amino acid changes within the ligand binding motifs. Despite evidence of the pseudogenization of various USP genes, we detected an important functional divergence at several residues, mostly located near sites that are critical for ligand interaction. CONCLUSIONS: Our results provide a broad framework for the evolution of the USP gene family, based on the emergence of new paralogs that face very contrasting fates, including pseudogenization, subfunctionalization or neofunctionalization. This framework aims to explain the sequence and functional diversity of this gene family, providing a foundation for future studies in other taxa in which USPs occur.

Increasing diversity of land planarians (Platyhelminthes: Geoplanidae) in the Interior Atlantic Forest with the description of two new species and new records from Argentina.

Here, we describe two new species of land planarians of the genera Pasipha and Imbira from the Atlantic Forest of Argentina: Pasipha quirogai sp. nov. and Imbira negrita sp. nov. Their external features and internal anatomy distinguish them from their congeners. Pasipha quirogai sp. nov. is about 40mm long, characterized by brown beige dorsal pigmentation with a golden yellow median band and thin graphite black para-median stripes, cylindrical pharynx, extrabulbar prostatic vesicle with paired proximal portion, male atrium with small folds in its proximal part, and female canal opening into the dorso-caudal wall of the female atrium. Imbira negrita sp. nov. is about 110mm long, with jet black dorsal pigmentation, pharynx collar-type, short female canal opening in the postero-dorsal portion of the female atrium, and epithelial lining of female atrium with stratified appearance. We also provide exhaustive descriptions of Geoplana quagga, Obama ladislavii, and Paraba multicolor, all recorded for the first time in Argentina, by analysing anatomical features of taxonomic relevance. In addition, we improve the description of G. quagga, mainly regarding the musculature, secretory cells, and the copulatory apparatus.

Conservation and diversification of small RNA pathways within flatworms.

BACKGROUND: Small non-coding RNAs, including miRNAs, and gene silencing mediated by RNA interference have been described in free-living and parasitic lineages of flatworms, but only few key factors of the small RNA pathways have been exhaustively investigated in a limited number of species. The availability of flatworm draft genomes and predicted proteomes allowed us to perform an extended survey of the genes involved in small non-coding RNA pathways in this phylum. RESULTS: Overall, findings show that the small non-coding RNA pathways are conserved in all the analyzed flatworm linages; however notable peculiarities were identified. While Piwi genes are amplified in free-living worms they are completely absent in all parasitic species. Remarkably all flatworms share a specific Argonaute family (FL-Ago) that has been independently amplified in different lineages. Other key factors such as Dicer are also duplicated, with Dicer-2 showing structural differences between trematodes, cestodes and free-living flatworms. Similarly, a very divergent GW182 Argonaute interacting protein was identified in all flatworm linages. Contrasting to this, genes involved in the amplification of the RNAi interfering signal were detected only in the ancestral free living species Macrostomum lignano. We here described all the putative small RNA pathways present in both free living and parasitic flatworm lineages. CONCLUSION: These findings highlight innovations specifically evolved in platyhelminths presumably associated with novel mechanisms of gene expression regulation mediated by small RNA pathways that differ to what has been classically described in model organisms. Understanding these phylum-specific innovations and the differences between free living and parasitic species might provide clues to adaptations to parasitism, and would be relevant for gene-silencing technology development for parasitic flatworms that infect hundreds of million people worldwide.

Three new species of Cratera Carbayo et al., 2013 from Araucaria forests with a key to species of the genus (Platyhelminthes, Continenticola).

Areas of Araucaria moist forest have been considered to constitute hotspots of land flatworm diversity, harbouring a high number of undescribed species. Herein we describe three new species of land flatworms of Cratera Carbayo et al., 2013 occurring in such type of forest in south Brazil. The three species are differentiated from their congeners mainly by their colour pattern, anatomy of the pharynx and prostatic vesicle, and details of the penis papilla and male atrium. An identification key to species of the genus in the Neotropical region is provided.

A new species of Notogynaphallia (Platyhelminthes, Geoplanidae) extends the known distribution of land planarians in Chacoan province (Chacoan subregion), South America.

BACKGROUND: The subfamily Geoplaninae (Geoplanidae) includes land planarian species of the Neotropical Region. In Argentina, the knowledge about land planarian diversity is still incipient, although this has recently increased mainly in the Atlantic Forest ecosystem. However, other regions like Chacoan forests remain virtually unexplored. RESULTS: In this paper, we describe a new species of the genus Notogynaphallia of the Chacoan subregion. This species is characterized by a black pigmentation on the dorsum and a dark grey ventral surface. The eyes with clear halos extend to the dorsal surface. The pharynx is cylindrical. The main features of the reproductive system involve testes anterior to the ovaries, prostatic vesicle intrabulbar (with a tubular proximal portion and a globose distal portion) opening broadly in a richly folded male atrium, common glandular ovovitelline duct and female genital canal dorso-anteriorly flexed constituting a "C",female atrium tubular proximally and widening distally. CONCLUSIONS: This is the first report of the genus Notogynaphallia in Argentina (Chacoan subregion, Neotropical Region) which increases its geographic distribution in South America. Also, as a consequence of features observed in species of the genus, we propose an emendation of the generic diagnosis.