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. Filariae are predominantly tissue-dwelling nematodes of the Onchocercidae (Spirurida). They are parasites of terrestrial vertebrates and some of them are particularly well known as agents of human diseases in tropical environments (e.g. onchocercosis, lymphatic filariosis, loaosis). Because of their predilection for host tissue sites, filariae are difficult to collect and their diversity is far from being well-investigated. Given the lack of fossilized material, it is not possible to formulate a comprehensive evolutionary hypothesis for this group based exclusively on morphological studies. Molecular analyses on the other hand have so far been based on a limited number of filarial species only or have used genetic markers convenient for species differentiation but not for retracing filarial history. Thus a consistent evolutionary framework for this family is still not in place. The originality and strength of the present study rest within its ability to access an unusually large range of biological samples, comprising 48 species representing seven of the eight onchocercid subfamilies. Using a new multi-gene dataset analysis, five major clades within the family were defined, including i) a group of ancestrally-derived subfamilies, ii) Setariinae as a sister group to all remaining subfamilies and iii) one large clade comprising genera of the Dirofilariinae, Onchocercinae and Splendidofilariinae.

Background: Among ocular vector-borne pathogens,Onchocerca volvulus, the agent of the so-called “river blindness”, affects about 37 million people globally. OtherOnchocercaspp. have been sporadically reported as zoonotic agents. Cases of canine onchocerciasis caused byOnchocerca lupiare on the rise in the United States and Europe. Its zoonotic role has been suspected but only recently ascertained in a single case from Turkey. The present study provides further evidence on the occurrence ofO. lupiinfesting human eyes in two patients from Turkey (case 1) and Tunisia (case 2). The importance of obtaining a correct sample collection and preparation of nematodes infesting human eyes is highlighted.Methods: In both cases the parasites were identified with morpho-anatomical characters at the gross examination, histological analysis and anatomical description and also molecularly in case 1.Results: The nematode from the first case was obviouslyO. lupibased on their morphology at the gross examination, histological analysis and anatomical description. In the second case, although the diagnostic cuticular characters were not completely developed, other features were congruent with the identification ofO. lupi. Furthermore, the morphological identification was also molecularly confirmed in the Turkish case.Conclusions: The results of this study suggest thatO. lupiinfestation is not an occasional finding but it should be considered in the differential diagnosis of other zoonotic helminths causing eye infestation in humans (e.g.,D. immitisandDirofilaria repens). Both cases came from areas where no cases of canine onchocerciasis were previously reported in the literature, suggesting that an in depth appraisal of the infestation in canine populations is necessary. Physicians and ophthalmologists are advised on how to preserve nematode samples recovered surgically, to allow a definitive, correct etiological diagnosis.

Based on material deposited in museum collections, twelve species within Mansonella sensu lato were examined and their descriptions amended. Based on additional morphological details, the erection of the new monotypic subgenus Filyamagutia Bain & Uni for M. (F.) akitensis (Uni, 1983), and the new combination M. (Pseudolitomosa) musasabi (Yamaguti, 1941) Bain & Uni are proposed. A new subspecies, M. (Tetrapetalonema) atelensis amazonae Bain & Guerrero is described and a key to the seven subgenera of Mansonella is provided. Furthermore, the elevation of Sandnema to full genus rank comprising the two species S. digitatum (Chandler, 1929) n. comb. and S. sunci (Sandground, 1933) n. comb., is proposed. Host and geographic records for the species of Mansonella and Sandnema are included.

SUMMARY: Aelurostrongylus abstrusus (Strongylida, Angiostrongylidae) and Troglostrongylus brevior (Strongylida, Crenosomatidae) are regarded as important lungworm species of domestic felids, with the latter considered an emerging threat in the Mediterranean region. The present study aimed to assess their concurrent development in the mollusc Helix aspersa (Pulmonata, Helicidae). Thirty snails were infested with 100 first-stage larvae (L1) of A. abstrusus and T. brevior, isolated from a naturally infested kitten. Larval development was checked by digesting five specimens at 2, 6 and 11 days post infestation. Larvae retrieved were morphologically described and their identification was confirmed by specific PCR and sequencing. All H. aspersa snails were positive for A. abstrusus and T. brevior, whose larval stages were simultaneously detected at each time point. In addition, snails were exposed to outdoor conditions and examined after overwintering, testing positive up to 120 days post infestation. Data herein presented suggest that A. abstrusus and T. brevior develop in H. aspersa snails and may eventually co-infest cats. Data on the morphology of both parasitic species in H. aspersa provide additional information on their development and identification, to better understand the population dynamics of these lungworms in receptive snails and paratenic hosts.