Background Wolbachia are intriguing symbiotic endobacteria with a peculiar host range that includes arthropods and a single nematode family, the Onchocercidae encompassing agents of filariases. This raises the question of the origin of infection in filariae. Wolbachia infect the female germline and the hypodermis. Some evidences lead to the theory that Wolbachia act as mutualist and coevolved with filariae from one infection event: their removal sterilizes female filariae; all the specimens of a positive species are infected; Wolbachia are vertically inherited; a few species lost the symbiont. However, most data on Wolbachia and filaria relationships derive from studies on few species of Onchocercinae and Dirofilariinae, from mammals.Methodology/Principal FindingsWe investigated the Wolbachia distribution testing 35 filarial species, including 28 species and 7 genera and/or subgenera newly screened, using PCR, immunohistochemical staining, whole mount fluorescent analysis, and cocladogenesis analysis. (i) Among the newly screened Onchocercinae from mammals eight species harbour Wolbachia but for some of them, bacteria are absent in the hypodermis, or in variable density. (ii) Wolbachia are not detected in the pathological model Monanema martini and in 8, upon 9, species of Cercopithifilaria. (iii) Supergroup F Wolbachia is identified in two newly screened Mansonella species and in Cercopithifilaria japonica. (iv) Type F Wolbachia infect the intestinal cells and somatic female genital tract. (v) Among Oswaldofilariinae, Waltonellinae and Splendidofilariinae, from saurian, anuran and bird respectively, Wolbachia are not detected.Conclusions/SignificanceThe absence of Wolbachia in 63% of onchocercids, notably in the ancestral Oswaldofilariinae estimated 140 mya old, the diverse tissues or specimens distribution, and a recent lateral transfer in supergroup F Wolbachia, modify the current view on the role and evolution of the endosymbiont and their hosts. Further genomic analyses on some of the newly sampled species are welcomed to decipher the open questions.

During the past twenty years, a number of molecular analyses have been performed to determine the evolutionary relationships of Onchocercidae, a family of filarial nematodes encompassing several species of medical or veterinary importance. However, opportunities for broad taxonomic sampling have been scarce, and analyses were based mainly on 12S rDNA and coxI gene sequences. While being suitable for species differentiation, these mitochondrial genes cannot be used to infer phylogenetic hypotheses at higher taxonomic levels. In the present study, 48 species, representing seven of eight subfamilies within the Onchocercidae, were sampled and sequences of seven gene loci (nuclear and mitochondrial) analysed, resulting in the hitherto largest molecular phylogenetic investigation into this family. Although our data support the current hypothesis that the Oswaldofilariinae, Waltonellinae and Icosiellinae subfamilies separated early from the remaining onchocercids, Setariinae was recovered as a well separated clade. Dirofilaria, Loxodontofilaria and Onchocerca constituted a strongly supported clade despite belonging to different subfamilies (Onchocercinae and Dirofilariinae). Finally, the separation between Splendidofilariinae, Dirofilariinae and Onchocercinae will have to be reconsidered.

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Angiostrongylus felineus n. sp. (Nematoda, Metastrongyloidea), parasitic in Puma (Herpailurus) yagouaroundi (É. Geoffroy, 1803) (Carnivora, Felidae) from the municipality of Juiz de Fora, Minas Gerais state, Brazil, is described and illustrated herein. Angiostrongylus felineus n. sp. differs from all congeneric species by having the anterior extremity with accentuated cuticular expansion and by smaller size of spicules. This study describes for the first time a species of Angiostrongylus in a wild Felidae in Brazil.

The helminth community of Tropidurus torquatus (Squamata: Tropiduridae) from a rocky outcrop area located in the state of Minas Gerais, southeastern Brazil, was studied. Ninety-two of the 110 individuals examined (83.6%) harbored helminths. Five species were found, including 3 nematodes ( Physaloptera lutzi , Parapharyngodon bainae , and Oswaldofilaria chabaudi ), 1 unidentified cestode species, and 1 acanthocephalan cystacanth also not identified. Only the nematode species had sufficient data to perform ecological analysis, with P. lutzi exhibiting the highest prevalence (67.3%). Prevalence between male and female hosts differed only for Oswaldofilaria chabaudi , with males exhibiting the highest values. The intensities of infection by P. lutzi and O. chabaudi were different among male and female hosts, with males also exhibiting the highest values. The host body size was positively related to intensity of infection for all nematode species. Local seasonality had some influence on the helminth community structure. Host diet, sexual dimorphism, and behavior (territorialism, forage strategy) represented important factors for the structure of this parasite community. In general, the helminth community was species poor, depauperate, and non-interactive, representing a typical structure observed in lizard hosts.