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      Molecular characterization of Wolbachia infection in bed bugs ( Cimex lectularius) collected from several localities in France Translated title: Caractérisation moléculaire de l’infection par Wolbachia chez les punaises de lit ( Cimex lectularius) recueillies dans plusieurs localités en France

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          Abstract

          Wolbachia symbionts are maternally inherited intracellular bacteria that have been detected in numerous insects including bed bugs. The objective of this study, the first epidemiological study in Europe, was to screen Wolbachia infection among Cimex lectularius collected in the field, using PCR targeting the surface protein gene ( wsp), and to compare obtained Wolbachia strains with those reported from laboratory colonies of C. lectularius as well as other Wolbachia groups. For this purpose, 284 bed bug specimens were caught and studied from eight different regions of France including the suburbs of Paris, Bouches-du-Rhône, Lot-et-Garonne, and five localities in Alpes-Maritimes. Among the samples, 166 were adults and the remaining 118 were considered nymphs. In all, 47 out of 118 nymphs (40%) and 61 out of 166 adults (37%) were found positive on wsp screening. Among the positive cases, 10 samples were selected randomly for sequencing. The sequences had 100% homology with wsp sequences belonging to the F-supergroup strains of Wolbachia. Therefore, we confirm the similarity of Wolbachia strains detected in this epidemiological study to Wolbachia spp. reported from laboratory colonies of C. lectularius.

          Translated abstract

          Les symbiotes Wolbachia sont des bactéries intracellulaires maternellement héritées qui ont été détectés chez de nombreux insectes, y compris les punaises de lit. Les objectifs de cette étude, la première étude épidémiologique en Europe, était de cribler l’infection par Wolbachia chez les Cimex lectularius recueillis sur le terrain, en utilisant la PCR ciblant le gène de la protéine de surface ( wsp) et de comparer la souche de Wolbachia obtenue avec celles rapportées des colonies de laboratoire de C. lectularius et d’autres groupes de Wolbachia. Dans ce but, 284 spécimens de punaises de lit ont été capturés et étudiés, provenant de huit régions différentes de la France, y compris les banlieues de Paris, les Bouches-du-Rhône, le Lot-et-Garonne et cinq localités dans les Alpes-Maritimes. Parmi les échantillons, 166 étaient des adultes et le reste (118) ont été considérés comme des nymphes. Quarante-sept des 118 nymphes (40 %) et 61 des 166 adultes (37 %) ont été trouvés positifs pour le dépistage de wsp. Parmi les cas positifs, 10 échantillons ont été choisis au hasard pour le séquençage. Les séquences avaient 100 % d’homologie avec des séquences de wsp appartenant aux souches du supergroupe F de Wolbachia. Par conséquent, nous confirmons la similitude des souches de Wolbachia détectées dans cette étude épidémiologique à celles des Wolbachia spp. signalées dans les colonies de laboratoire de C. lectularius.

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          Multilocus sequence typing system for the endosymbiont Wolbachia pipientis.

          The eubacterial genus Wolbachia comprises one of the most abundant groups of obligate intracellular bacteria, and it has a host range that spans the phyla Arthropoda and Nematoda. Here we developed a multilocus sequence typing (MLST) scheme as a universal genotyping tool for Wolbachia. Internal fragments of five ubiquitous genes (gatB, coxA, hcpA, fbpA, and ftsZ) were chosen, and primers that amplified across the major Wolbachia supergroups found in arthropods, as well as other divergent lineages, were designed. A supplemental typing system using the hypervariable regions of the Wolbachia surface protein (WSP) was also developed. Thirty-seven strains belonging to supergroups A, B, D, and F obtained from singly infected hosts were characterized by using MLST and WSP. The number of alleles per MLST locus ranged from 25 to 31, and the average levels of genetic diversity among alleles were 6.5% to 9.2%. A total of 35 unique allelic profiles were found. The results confirmed that there is a high level of recombination in chromosomal genes. MLST was shown to be effective for detecting diversity among strains within a single host species, as well as for identifying closely related strains found in different arthropod hosts. Identical or similar allelic profiles were obtained for strains harbored by different insect species and causing distinct reproductive phenotypes. Strains with similar WSP sequences can have very different MLST allelic profiles and vice versa, indicating the importance of the MLST approach for strain identification. The MLST system provides a universal and unambiguous tool for strain typing, population genetics, and molecular evolutionary studies. The central database for storing and organizing Wolbachia bacterial and host information can be accessed at http://pubmlst.org/wolbachia/.
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            Beyond insecticides: new thinking on an ancient problem.

            Vector-borne disease is one of the greatest contributors to human mortality and morbidity throughout the tropics. Mosquito-transmitted diseases such as malaria, dengue, yellow fever and filariasis are the main contributors to this burden. Although insecticides have historically been used to try to control vector populations, over the past 15 years, substantial progress has been made in developing alternative vector control strategies ranging from biocontrol methods through to genetic modification of wild insect populations. Here, we review recent advances concerning these strategies and consider the potential impediments to their deployment, including the challenges of obtaining regulatory approval and community acceptance.
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              Bad guys turned nice? A critical assessment of Wolbachia mutualisms in arthropod hosts.

              Wolbachia are the most abundant bacterial endosymbionts among arthropods. Although maternally inherited, they do not conform to the widespread view that vertical transmission inevitably selects for beneficial symbionts. Instead, Wolbachia are notorious for their reproductive parasitism which, although lowering host fitness, ensures their spread. However, even for reproductive parasites it can pay to enhance host fitness. Indeed, there is a recent upsurge of reports on Wolbachia-associated fitness benefits. Therefore, the question arises how such instances of mutualism are related to the phenotypes of reproductive parasitism. Here, we review the evidence of Wolbachia mutualisms in arthropods, including both facultative and obligate relationships, and critically assess their biological relevance. Although many studies report anti-pathogenic effects of Wolbachia, few actually prove these effects to be relevant to field conditions. We further show that Wolbachia frequently have beneficial and detrimental effects at the same time, and that reproductive manipulations and obligate mutualisms may share common mechanisms. These findings undermine the idea of a clear-cut distinction between Wolbachia mutualism and parasitism. In general, both facultative and obligate mutualisms can have a strong, and sometimes unforeseen, impact on the ecology and evolution of Wolbachia and their arthropod hosts. Acknowledging this mutualistic potential might be the key to a better understanding of some unresolved issues in the study of Wolbachia-host interactions.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2016
                05 August 2016
                : 23
                : ( publisher-idID: parasite/2016/01 )
                : 31
                Affiliations
                [1 ] Service de Parasitologie-Mycologie, Hôpital de l’Archet, Centre Hospitalier Universitaire de Nice 06000 Nice France
                [2 ] Inserm U1065, Centre Méditerranéen de Médecine Moléculaire, Faculté de Médecine, Université Nice-Sophia Antipolis 06000 Nice France
                [3 ] Faculté de Médecine, Université Nice-Sophia Antipolis 06000 Nice France
                [4 ] URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, Faculté de Médecine, Université Aix-Marseille 13000 Marseille France
                [5 ] Service de Parasitologie-Mycologie, Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris 93000 Bobigny France
                Author notes
                [* ]Corresponding author: m.akhoundi@ 123456yahoo.com
                Article
                parasite160016 10.1051/parasite/2016031
                10.1051/parasite/2016031
                4974871
                27492563
                bc0ec827-de2f-46c0-92c8-cd3110cd3c1b
                © M. Akhoundi et al., published by EDP Sciences, 2016

                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.

                History
                : 28 February 2016
                : 23 July 2016
                Page count
                Figures: 2, Tables: 1, Equations: 0, References: 37, Pages: 6
                Categories
                Research Article

                bed bugs,cimex lectularius,symbiont,wolbachia
                bed bugs, cimex lectularius, symbiont, wolbachia

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