Morphology, complete mitochondrial genome, and molecular phylogeny of Rhabdias macrocephalum n. sp. (Nematoda: Rhabdiasidae) from Diploderma splendidum (Reptilia: Agamidae).

Species of the genus Rhabdias Stiles & Hassall, 1905 are common parasitic nematodes occurring in the lungs of amphibians and reptiles worldwide. In the present study, Rhabdias macrocephalum n. sp. is described using integrated morphological methods (light and scanning electron microscopy) and molecular approaches (sequencing of the nuclear 28S and ITS regions, and mitochondrial cox1, cox2, and 12S genes) based on specimens collected from the green striped tree dragon Diploderma splendidum (Barbour & Dunn) (Reptilia: Agamidae) in China. The complete mitochondrial genome of R. macrocephalum n. sp. was sequenced and annotated: it is 14,819 bp in length, including 12 protein coding genes (missing atp8), 22 tRNA genes, 2 rRNA genes and three non-coding regions. The gene arrangement of R. macrocephalum n. sp. is different from all of the currently available mitogenomes of nematodes and represents a novel type of mitochondrial gene arrangement reported in Nematoda. Molecular phylogenetic results based on the ITS + 28S data support the monophyly of Entomelas, Pneumonema, Serpentirhabdias, and Rhabdias, and showed R. macrocephalum n. sp. forming a most basal lineage in Rhabdias.

Title: Morphologie, génome mitochondrial complet et phylogénie moléculaire de Rhabdias macrocephalum n. sp. (Nematoda : Rhabdiasidae) de Diploderma splendidum (Reptilia : Agamidae). Abstract: Les espèces du genre Rhabdias Stiles & Hassall, 1905 sont des nématodes parasites courants présents dans les poumons des amphibiens et des reptiles du monde entier. Dans cette étude, Rhabdias macrocephalum n. sp. est décrit à l'aide de méthodes morphologiques intégrées (microscopie optique et électronique à balayage) et d'approches moléculaires (séquençage des régions nucléaires 28S et ITS et des gènes mitochondriaux cox1, cox2 et 12S) basées sur des spécimens collectés chez le lézard Diploderma splendidum (Barbour & Dunn) (Reptilia : Agamidae) de Chine. Le génome mitochondrial complet de R. macrocephalum n. sp. a été séquencé et annoté : il a une longueur de 14 819 pb, dont 12 gènes codants pour des protéines (atp8 manquant), 22 gènes d'ARNt, 2 gènes d'ARNr et trois régions non codantes. L'arrangement génétique de R. macrocephalum n. sp. est différent de tous les mitogénomes de nématodes actuellement disponibles et représente un nouveau type d'arrangement de gènes mitochondriaux signalé chez les nématodes. Les résultats phylogénétiques moléculaires basés sur les données ITS + 28S ont soutenu la monophylie d'Entomelas, Pneumonema, Serpentirhabdias et Rhabdias, et ont montré que R. macrocephalum n. sp. forme la lignée la plus basale chez Rhabdias.

DESCRIPTIONS AND PHYLOGENETIC AFFINITIES OF TWO NEW SPECIES OF RHABDIAS STILES AND HASSALL, 1905 (NEMATODA: RHABDIASIDAE) FROM NORTH AMERICAN FROGS: ARE WE STILL ONLY SCRATCHING THE SURFACE?

Two new species of lung-dwelling nematodes are described from North American frogs: Rhabdias aurorae n. sp. from Rana aurora and Rhabdias conni n. sp. from Rana clamitans and Rana catesbeiana from Arkansas; the latter species was also found in Oklahoma and Georgia. Rhabdias aurorae n. sp. differs from other Nearctic congeners in the combination of the following characteristics: buccal capsule 22-25 µm wide, elongated tail covered with inflated cuticle, esophagus with prominent dilatation in anterior part and 6 small circumoral lips. Rhabdias conni n. sp. is morphologically closest to Rhabdias ranae Walton, 1929 and Rhabdias joaquinensisIngles, 1936; it differs from them in the shape of lateral pseudolabia, the dimensions of the body, and the egg size. Both new species were found to be significantly different from the Nearctic congeners in the nucleotide sequences of nuclear ribosomal DNA (18S-ITS-28S region), 12S, and CO1 mitochondrial genes. The 2 new species differ from other currently sequenced Nearctic congeners by 1.1-2.7% of nucleotide positions in the nuclear rDNA region, 1.3-3.4% in the 12S gene, and 3.4-9.4% in CO1 gene. Molecular phylogenetic analysis based on nuclear ribosomal DNA sequences placed both new species into the clade consisting of Nearctic and Neotropical Rhabdias spp. The position of Rh. aurorae n. sp. within the clade is uncertain because of a polytomy, but Rh. conni n. sp. is nested within the "Rh. joaquinensis complex" related to Rh. ranae and Rhabdias tarichaeKuzmin, Tkach, and Snyder, 2003. The phylogenetic analysis based on nuclear ribosomal DNA sequences has revealed 3 evolutionary host-switching events from anuran to caudatan hosts among Rhabdias spp. that occurred in the Nearctic and Palearctic. The molecular phylogeny also suggests that Rhabdias may have originally evolved in what is now Africa.

Variability study of entomopathogenic nematode populations (Heterorhabditidae) from Argentina / Estudo da variabilidade entomopatogênicos nematóides populações (Heterorhabditidae) da Argentina

Abstract Entomopathogenic nematodes (EPN) belonging to the Heterorhabditidae family are lethal parasites of soil-dwelling insects. Two species were reported in Argentina: Heterorhabditis argentinensis and Heterorhabditis bacteriophora characterized mainly by morphometric features. In this work a comparative and phylogenetic study between five Heterorhabditis populations from Argentina was conducted to analyze the variability between strains and to evaluate the taxonomic position of Heterorhabditis argentinensis. The PCA analyses of morphometric characters separated the larger juvenile, female and male H. argentinensis from H. bacteriophora populations. The juvenile (IJs) stage provided the clearest separation of Heterorhabditis populations presenting the least variability between strains. The variable L and MBW were highly related to H. argentinensis IJs. Three groups were separated by this stage considering PC1 and PC2: one formed by H. bacteriophora OLI, RIV and RN strains, (isolates from Córdoba and Río Negro province), one for H. bacteriophora VELI strain (Buenos Aires province) and one for H. argentinensis (Santa Fe province). Heterorhabditis bacteriophora VELI and H. argentinensis isolated from regions with more rainfalls and humidity presented larger values for morphometric features. Molecular analyses showed the Argentinian populations (H. bacteriophora VELI strain and H. argentinensis), forming a same clade, with six other H. bacteriophora populations (not from Argentina) with a genetic similarity between them of 99%. Heterorhabditis argentinensis presented one unique nucleotide that was not present in any of the other species of the clade. Considering the results of this study H. argentinensis would be conspecific to H. bacteriophora, constituting a strain with a great morphometric variation where the host and climatic conditions could have influenced on the measurements.

Resumo Nematóides entomopatogênicos (EPN) pertencentes à família Heterorhabditidae são parasitas letais de insetos que vivem no solo. Duas espécies foram relatados na Argentina: Heterorhabditis argentinensis e Heterorhabditis bacteriophora, caracterizada principalmente por características morfométricas. Neste trabalho um estudo comparativo e filogenética entre cinco populações do Heterorhabditis da Argentina foi conduzido para analisar a variabilidade entre as linhagens e avaliar a posição taxonômica das Heterorhabditis argentinensis. Características morfométricas de Heterorhabditis bacteriophora VELI e H. argentinensis isoladas de regiões com mais chuvas e umidade apresentaram dimensões maiores. Analisa o PCA de personagens morfométricas separou a maior juvenil, feminino e masculino H. argentinensis de H. bacteriophora populações. A fase juvenil (JIs) fornece a mais clara separação de populações Heterorhabditis apresentando a menor variação entre as cepas. A L variável e MBW foram altamente relacionada com H. argentinensis JIs. Três grupos foram separados por esta fase considerando PC1 e PC2: um formado por H. bacteriophora OLI, RIV e estirpes RN, (isolados de Córdoba e província de Rio Negro), um para a estirpe H. bacteriophora VELI (província de Buenos Aires) e um para H. argentinensis (província de Santa Fe). Heterorhabditis bacteriophora VELI e H. argentinensis isolado a partir de regiões com mais chuvas e umidade apresentaram maiores valores para as características morfométricas. A análise molecular mostrou as populações da Argentina (estirpe H. bacteriophora VELI e H. argentinensis), formando um mesmo subtipo, com seis outras populações H. bacteriophora (não da Argentina), com uma similaridade genética entre eles de 99%. Heterorhabditis argentinensis apresentado um único nucleótido que não estava presente em nenhum dos outros espécies do clado. Considerando os resultados deste estudo H. argentinensis seria conspecific a H. bacteriophora, constituindo uma estirpe com uma grande variação morfométrica onde o anfitrião e as condições climáticas podem ter influenciado nas medições.

Um caso de parasitismo por Rhabditis sp em criança natural de Goiânia, Goiás, Brasil / A case of parasitism by Rhabditis sp in a child from Goiânia, Goiás, Brazil

Relata-se um caso de parasitismo por Rhabditis sp em criança com cinco meses de idade procedente do Estado de Goiás, Brasil. O quadro clínico mostrou inicialmente diarréia com fezes líquidas esverdeadas e posteriormente sanguinolentas. O exame parasitológico de fezes revelou a presença de larvas e adultos de Rhabditis sp. Após o uso de thiabendazole houve melhora no quadro clínico e cura. Os autores chamam atençäo para a importância de se estabelecer o diagnóstico diferencial entre Strongyloides e Rhabditis