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      Infective larvae of Cercopithifilaria spp. (Nematoda: Onchocercidae) from hard ticks (Ixodidae) recovered from the Japanese serow (Bovidae) Translated title: Larves infectantes de Cercopithifilaria spp. (Nematoda: Onchocercidae) chez des tiques (Ixodidae) récoltées sur des sérows japonais (Bovidae)

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          Abstract

          Hard ticks taken from the Japanese serow, Capricornis crispus, in Yamagata Prefecture, Honshu, harboured infective larvae of onchocercid filariae after incubation from the 22nd to the 158th day. Haemaphysalis flava and H. japonica contained one to eight filarial larvae; females, males and a nymph of the ticks were infected. The 44 infective larvae recovered were 612–1,370 μm long, and 11 of them, 930–1,340 μm long, were studied in detail. The larvae possessed the morphologic characteristics of the larvae of the genus Cercopithifilaria, namely an oesophagus with a posterior glandular part, no buccal capsule and a long tail with three terminal lappets. Five types (A to E) of infective larvae were identified based on the morphologic characteristics. While to date five species of Cercopithifilaria have been described from the Japanese serow, a specific identification of the larvae found in this study was generally not possible. Only type E larvae could be tentatively assigned to Cercopithifilaria tumidicervicata, as they had a cervical swelling similar to that of the adults of this species. A key for the identification of the five larval types is presented. The study presents circumstantial evidences indicating that H. flava and H. japonica may transmit Cercopithifilaria spp. to Japanese serows. It also suggests the possibility that such filarial larvae will be found in hard ticks anywhere, because Cercopithifilaria is distributed worldwide, though this genus generally goes unnoticed, as its microfilariae occur in the skin, not in the blood, of host animals.

          Translated abstract

          Les tiques récoltées chez les sérows japonais, Capricornis crispus, à Yamagata, Honshu, hébergeaient des larves infectantes de filaires Onchocercidae après incubation du 22 ème au 158 ème jour. Haemaphysalis flava et H. japonica avaient une à huit larves ; des mâles, des femelles et une nymphe étaient infestés. Les 44 larves infectantes récoltées étaient longues de 612–1,370 μm et 11 d’entre elles, longues de 930 à 1,340 μm, ont été étudiées en détail. Les larves possédaient les caractères morphologiques des larves du genre Cercopithifilaria, c’est-à-dire un œsophage avec une portion postérieure glandulaire, pas de capsule buccale et une longue queue avec 3 languettes terminales. Cinq types de larves infectantes (A à E) ont été identifiés sur la base de caractéristiques morphologiques. Bien que cinq espèces de Cercopithifilaria aient été décrites jusqu’ici chez le sérow japonais, une identification spécifique des larves trouvées dans cette étude n’a généralement pas été possible. Seules les larves de type E ont été identifiées provisoirement à C. tumidicervicata parce qu’elles avaient un renflement cervical comme les adultes de cette espèce. Une clé d’identification des cinq types de larves est présentée. L’étude présente des preuves circonstancielles indiquant que H. flava et H. japonica peuvent transmettre Cercopithifilaria spp. au sérow japonais. Elle suggère la possibilité de trouver partout de telles larves de filaires chez les tiques parce que Cercopithifilaria a une distribution mondiale mais passe généralement inaperçu parce que ses microfilaires sont dans la peau, et non dans le sang.

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          Integrated taxonomy: traditional approach and DNA barcoding for the identification of filarioid worms and related parasites (Nematoda)

          Background We compared here the suitability and efficacy of traditional morphological approach and DNA barcoding to distinguish filarioid nematodes species (Nematoda, Spirurida). A reliable and rapid taxonomic identification of these parasites is the basis for a correct diagnosis of important and widespread parasitic diseases. The performance of DNA barcoding with different parameters was compared measuring the strength of correlation between morphological and molecular identification approaches. Molecular distance estimation was performed with two different mitochondrial markers (coxI and 12S rDNA) and different combinations of data handling were compared in order to provide a stronger tool for easy identification of filarioid worms. Results DNA barcoding and morphology based identification of filarioid nematodes revealed high coherence. Despite both coxI and 12S rDNA allow to reach high-quality performances, only coxI revealed to be manageable. Both alignment algorithm, gaps treatment, and the criteria used to define the threshold value were found to affect the performance of DNA barcoding with 12S rDNA marker. Using coxI and a defined level of nucleotide divergence to delimit species boundaries, DNA barcoding can also be used to infer potential new species. Conclusion An integrated approach allows to reach a higher discrimination power. The results clearly show where DNA-based and morphological identifications are consistent, and where they are not. The coherence between DNA-based and morphological identification for almost all the species examined in our work is very strong. We propose DNA barcoding as a reliable, consistent, and democratic tool for species discrimination in routine identification of parasitic nematodes.
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            Morphological and molecular data on the dermal microfilariae of a species of Cercopithifilaria from a dog in Sicily.

            Dermal microfilariae found in a dog from Sicily, Italy, were characterized morphologically and genetically and differentiated from those of all the other blood microfilariae commonly found in dogs. In particular, the microfilariae were short (mean length of 186.7 μm), presented a body flattened dorso-ventrally and a rounded head, bearing a tiny cephalic hook. The genetic identity of microfilariae herein studied was also assessed by molecular amplification, sequencing and analyzing of multiple ribosomal ITS-2 and mitochondrial (cox1 and 12S) target genes. Both morphologic and genetic characterization as well as the molecular phylogenetic history inferred using sequences of a barcoding dataset were concordant in supporting the identification of Cercopithifilaria at the genus level. Surprisingly, microfilariae here examined were well distinct from Cercopithifilaria grassii (Noè, 1907), from northern Italy, and resembled those of a species described in Brazil, Cercopithifilaria bainae Almeida & Vicente, 1984. This paper provides evidence for the existence of a Cercopithifilaria species infesting a dog from Sicily and also presents a PCR protocol on skin samples as a tool for further epidemiological studies, which could provide evidence on the aetiology and the natural history of this filarial species. Copyright © 2011 Elsevier B.V. All rights reserved.
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              Rhipicephalus sanguineus (Ixodida, Ixodidae) as intermediate host of a canine neglected filarial species with dermal microfilariae.

              The life cycles of filarioids of dogs presenting dermal microfilariae have been little studied. Following the recent retrieval of dermal microfilariae identified as Cercopithifilaria sp. in a dog from Sicily (Italy), this study was designed to assess the role of the brown dog tick Rhipicephalus sanguineus as an intermediate host of this filarial species. An experimental tick infestation was performed on an infected dog using 300 nymphs of R. sanguineus. Engorged nymphs were collected and examined by both microscopic dissection and molecular analysis at five time points (i.e., the same day of tick detachment and 10, 20, 30 and 50 days post-detachment) to detect the presence and developmental stage of filariae in the ticks. A total of 270 engorged nymphs were collected from the dog and developing filarioid larvae detected in 10 (5%) out of 200 ticks dissected. Infective third-stage larvae were observed in 4 (2%) of the all dissected ticks, 30 days post-detachment. Twelve (6.6%) out of 181 samples molecularly tested were positive for Cercopithifilaria sp. This study demonstrates that nymphs of R. sanguineus feeding on a dog naturally infected by Cercopithifilaria sp. can ingest microfilariae, which develop up to the third infective stage thus suggesting that this tick species might act as an intermediate host of this little known canine filarioid. Copyright © 2011 Elsevier B.V. All rights reserved.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2013
                17 January 2013
                : 20
                : ( publisher-idID: parasite/2013/01 )
                : 1
                Affiliations
                [1 ] Institute of Biological Sciences, Faculty of Science, University of Malaya 50603 Kuala Lumpur Malaysia
                [2 ] Department of Parasitology, Graduate School of Medicine, Osaka City University Osaka 545-8585 Japan
                [3 ] Muséum National d’Histoire Naturelle, Parasitologie comparée, UMR 7205, CNRS 75231 Paris France
                [4 ] Mahara Institute of Medical Acarology Tokushima 779-1510 Japan
                [5 ] National Institute of Animal Health, NARO Ibaraki 305-0856 Japan
                [6 ] Research Promotion Project, Oita University Oita 879-5593 Japan
                [7 ] Department of Infectious Disease Control, Faculty of Medicine, Oita University Oita 879-5593 Japan
                Author notes
                [* ]Corresponding author: unishigehiko@ 123456um.edu.my
                Article
                parasite120011 10.1051/parasite/2012001
                10.1051/parasite/2012001
                3718534
                23340227
                0b4469d0-2966-49b1-b0b4-824e2f64abf8
                © S. Uni et al., published by EDP Sciences, 2013

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 12 September 2012
                : 19 November 2012
                Page count
                Figures: 1, Tables: 1, Equations: 0, References: 39, Pages: 7
                Categories
                Research Article

                nematoda,onchocercidae,cercopithifilaria,ixodidae,infective larvae,japanese serow

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