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      Molecular analysis of Dirofilaria repens removed from a subcutaneous nodule in a Japanese woman after a tour to Europe Translated title: Analyse moléculaire de Dirofilaria repens retiré d’un nodule sous-cutané chez une femme japonaise après un voyage en Europe

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          Abstract

          A premature female Dirofilaria species, subsequently identified as Dirofilaria repens by its morphological features and mitochondrial 12S ribosomal RNA ( 12S rRNA) gene sequence, was removed from a subcutaneous nodule of the right temporal region of the head in a Japanese woman 2 years after she noticed swelling of her left calf following an insect sting during a tour to Europe; headache symptoms were noticed a few months later. The sequences of the mitochondrial 12S rRNA and cytochrome c oxidase subunit I genes from the organism were almost identical to those of sequences AM779772 (100% homology, 337/337) and AM749233 (99.8% homology, 536/537) of D. repens isolated from humans in Italy. However, the phylogenetic position of the 18S rRNA-internal transcribed spacer 1-5.8S rRNA region was in the same cluster as that of sequence JX290195 of Dirofilaria sp. “ hongkongensis” (96.7% homology, 348/360), which was recently reported from Hong Kong as a novel Dirofilaria species. Information on regional genetic variation in D. repens isolated from animals and humans remains scarce. We report the detailed genetic features of this filaria as a reference isolate from a specific endemic area, to enrich the genetic database of D. repens.

          Translated abstract

          Une femelle immature de Dirofilaria, par la suite identifiée comme Dirofilaria repens par ses caractéristiques morphologiques et la séquence du gène de son ARN ribosomique mitochondrial 12S (ARNr 12S), a été retirée d’un nodule sous-cutané de la région temporale droite de la tête d’une femme japonaise, deux ans après qu’elle ait remarqué un gonflement de son mollet gauche suite à une piqûre d’insecte lors d’un voyage d’agrément en Europe. Des symptômes de maux de tête ont été remarqués quelques mois plus tard. Les séquences des gènes de l’ARNr mitochondrial 12S et de la sous-unité I de la cytochrome c oxydase de l’organisme étaient presque identiques à celles des séquences AM779772 (100 % d’homologie, 337/337) et AM749233 (99,8 % d’homologie, 536/537) de D. repens, isolées chez l’homme en Italie. Cependant, la position phylogénétique de la région intercalaire 1-5.8S de l’ARNr 18S était dans le même groupe que celui de la séquence JX290195 de Dirofilaria sp. “ hongkongensis” (96.7 % d’homologie, 348/360), qui a été récemment rapporté à Hong Kong comme une nouvelle espèce de Dirofilaria. Les informations sur la variation génétique régionale de D. repens isolés chez les animaux et les humains restent rares. Nous rapportons les caractéristiques génétiques détaillées de cette filaire comme isolat de référence d’une zone endémique spécifique, pour enrichir la base de données génétique de D. repens.

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          Most cited references 18

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          Vector-borne helminths of dogs and humans in Europe

          Presently, 45% of the total human population of Europe, as well as their domestic and companion animals, are exposed to the risk of vector-borne helminths (VBH) causing diseases. A plethora of intrinsic biological and extrinsic factors affect the relationship among helminths, vectors and animal hosts, in a constantly changing environment. Although canine dirofilarioses by Dirofilaria immitis and Dirofilaria repens are key examples of the success of VBH spreading into non-endemic areas, another example is represented by Thelazia callipaeda eyeworm, an emergent pathogen of dogs, cats and humans in several regions of Europe. The recent finding of Onchocerca lupi causing canine and human infestation in Europe and overseas renders the picture of VBH even more complicated. Similarly, tick-transmitted filarioids of the genus Cercopithifilaria infesting the skin of dogs were recently shown to be widespread in Europe. Although for most of the VBH above there is an increasing accumulation of research data on their distribution at national level, the overall impact of the diseases they cause in dogs and humans is not fully recognised in many aspects. This review investigates the reasons underlying the increasing trend in distribution of VBH in Europe and discusses the diagnostic and control strategies currently available. In addition, this article provides the authors’ opinion on some topics related to VBH that would deserve further scientific investigation.
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            Dirofilaria repens Infection and Concomitant Meningoencephalitis

            Dirofilaria repens, a filarial nematode of dogs and other carnivores, can accidentally infect humans. Clinical symptoms are usually restricted to a subcutaneous nodule containing a single infertile parasite. Here, we report a case of D. repens infection with a subcutaneous gravid worm and the patient’s concomitant meningoencephalitis and aphasia.
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              Detection and differentiation of filarial parasites by universal primers and polymerase chain reaction-restriction fragment length polymorphism analysis.

              Filarial nematode parasites are a serious cause of morbidity in humans and animals. Identification of filarial infection using traditional morphologic criteria can be difficult and lead to misdiagnosis. We report on a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based method to detect and differentiate a broad range of filarial species in a single PCR. The first internal transcribed spacer 1 (ITS1) along with the flanking 18S and 5.8S ribosomal DNA (rDNA) were isolated and cloned from Wuchereria bancrofti, Brugia malayi, and Brugia pahangi. Sequence analysis identified conserved sites in the 18S and 5.8S rDNA sequence that could be used as universal priming sites to generate ITS1-distinctive PCR products that were useful for distinguishing filariae at the genus level. The addition of a digestion of the ITS1 PCR product with the restriction endonuclease Ase I generated a fragment profile that allowed differentiation down to the species level for W. bancrofti, B. malayi, B. pahangi, Dirofilaria immitis, and D. repens. The PCR-RFLP of ITS1 rDNA will be useful in diagnosing and differentiating filarial parasites in human, animal reservoir hosts, and mosquito vectors in disease-endemic areas.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2015
                27 January 2015
                : 22
                : ( publisher-idID: parasite/2015/01 )
                Affiliations
                [1 ] Division of Clinical Microbiology, Department of Microbiology, Tokyo Metropolitan Institute of Public Health 3-24-1 Hyakunincho Shinjuku-ku, Tokyo 169-0073 Japan
                [2 ] Department of Infectious Diseases, Keio University School of Medicine 35 Shinanomachi Shinjuku-ku, Tokyo 160-8582 Japan
                [3 ] Department of Dermatology, Saiseikai Central Hospital 1-4-17 Mita Minato-ku, Tokyo 108-0073 Japan
                [4 ] Department of Dermatology, Keio University School of Medicine 35 Shinanomachi Shinjuku-ku, Tokyo 160-8582 Japan
                Author notes
                [* ]Corresponding author: skobaya@ 123456z7.keio.jp
                Article
                parasite140076 10.1051/parasite/2015002
                10.1051/parasite/2015002
                4306022
                25619827
                © J. Suzuki et al., published by EDP Sciences, 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 2, Tables: 3, Equations: 0, References: 28, Pages: 8
                Categories
                Research Article

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