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Broader prevalence of Wolbachia in insects including potential human disease vectors

  • C.D. de Oliveira (a1), D.S. Gonçalves (a1), L.A. Baton (a1), P.H.F. Shimabukuro (a2), F.D. Carvalho (a1) and L.A. Moreira (a1)...


Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.


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Almerão, M.P., Fagundes, N.J.R., de Araújo, P.B., Verne, S., Grandjean, F., Bouchon, D. & Araújo, A.M. (2012) First record of Wolbachia in South American terrestrial isopods: prevalence and diversity in two species of Balloniscus (Crustacea, Oniscidea). Genetics and Molecular Biology 35, 980989.
Augustinos, A.A., Santos-Garcia, D., Dionyssopoulou, E., Moreira, M., Papapanagiotou, A., Scarvelakis, M., Doudoumis, V., Ramos, S., Aguiar, A.F., Borges, P.A., Khadem, M., Latorre, A., Tsiamis, G. & Bourtzis, K. (2011) Detection and characterization of Wolbachia infections in natural populations of aphids: is the hidden diversity fully unraveled? PLoS ONE 6, e28695.
Azpurua, J., De La Cruz, D., Valderama, A. & Windsor, D. (2010) Lutzomyia sand fly diversity and rates of infection by Wolbachia and an exotic Leishmania species on Barro Colorado Island, Panama. PLoS Neglected Tropical Diseases 4, e627.
Baldini, F., Segata, N., Pompon, J., Marcenac, P., Robert Shaw, W., Dabiré, R.K., Diabaté, A., Levashina, E.A. & Catteruccia, F. (2014). Evidence of natural Wolbachia infections in field populations of Anopheles gambiae . Nature Communications 5, 3985.
Bandi, C., Sironi, M., Nalepa, C.A., Corona, S. & Sacchi, L. (1997) Phylogenetically distant intracellular symbionts in termites. Parassitologia 39, 7175.
Baton, L.A., Pacidônio, E.C., da Silva Gonçalves, D. & Moreira, L.A. (2013) wFlu: characterization and evaluation of a native Wolbachia from the mosquito Aedes fluviatilis as a potential vector control agent. PLoS ONE 8, e59619.
Benlarbi, M. & Ready, P.D. (2003) Host-specific Wolbachia strains in widespread populations of Phlebotomus perniciosus and P. papatasi (Diptera: Psychodidae), and prospects for driving genes into these vectors of Leishmania . Bulletin of Entomological Research 93, 383391.
Bordenstein, S. & Rosengaus, R.B. (2005) Discovery of a novel Wolbachia super group in Isoptera. Current microbiology 51, 393398.
Boyle, L., O'Neill, S.L., Robertson, H.M. & Karr, T.L. (1993) Interspecific and intraspecific horizontal transfer of Wolbachia in Drosophila . Science 260, 17961799.
Braig, H.R., Guzman, H., Tesh, R.B. & O'Neill, S.L. (1994) Replacement of the natural Wolbachia symbiont of Drosophila simulans with a mosquito counterpart. Nature 367, 453455.
Brucker, R.M. & Bordenstein, S.R. (2012) Speciation by symbiosis. Trends in Ecology and Evolution 27, 443451.
Casiraghi, M., Bordenstein, S.R., Baldo, L., Lo, N., Beninati, T., Wernegreen, J.J., Werren, J.H. & Bandi, C. (2005) Phylogeny of Wolbachia pipientis based on gltA, groEL and ftsZ gene sequences: clustering of arthropod and nematode symbionts in the F supergroup, and evidence for further diversity in the Wolbachia tree. Microbiology 151, 40154022.
Consoli, R.A.G.B. & de Oliveira, R.L. (1994) Principais Mosquitos de Importância Sanitária no Brasil. p. 228. Rio de Janeiro, Fiocruz.
Costa, J. & Lorenzo, M. (2009) Biology, diversity and strategies for the monitoring and control of triatomines – Chagas disease vectors. Memorias do Instituto Oswaldo Cruz 104, 4651.
Cui, L., Chang, S.H., Strickman, D. & Rowton, E. (1999) Frequency of Wolbachia infection in laboratory and field sand fly (Diptera: Psychodidae) populations. Journal of the American Mosquito Control Association 15, 571572.
de Albuquerque, A.L., Magalhães, T. & Ayres, C.F.J. (2011) High prevalence and lack of diversity of Wolbachia pipientis in Aedes albopictus populations from Northeast Brazil. Memorias Instituto Oswaldo Cruz 106, 773776.
de Almeida, F., Moura, A.S., Cardoso, A.F., Winter, C.E., Bijovsky, A.T. & Suesdek, L. (2011) Effects of Wolbachia on fitness of Culex quinquefasciatus (Diptera; Culicidae). Infection, Genetics and Evolution 11, 21382143.
Dean, J.L. & Dobson, S.L. (2004) Characterization of Wolbachia infections and interspecific crosses of Aedes (Stegomyia) polynesiensis and Ae. (Stegomyia) riversi (Diptera: Culicidae). Journal of Medical Entomology 41, 894900.
de Sousa, K.B.A., da Silva, T.R.R., Alencar, R.B., Baton, L.A., Naveca, F.G. & Shimabukuro, P.H.F. (2013) 16S rRNA gene-based identification of microbiota associated with the parthenogenetic troglobiont sand fly Deanemyia maruaga (Diptera, Psychodidae) from central Amazon, Brazil. Brazilian Journal of Microbiology 44, 325328.
Dittmar, K. & Whiting, M.F. (2004) New Wolbachia endosymbionts from Nearctic and Neotropical fleas (Siphonaptera). Journal of Parasitology 90, 953957.
Dong, P., Wang, J-J. & Zhao, Z-M. (2006) Infection by Wolbachia bacteria and its influence on the reproduction of the stored-product psocid, Liposcelis tricolor . Journal of Insect Science 6, 17.
Dumler, J.S., Barbet, A.F., Bekker, C.P.J., Dasch, G.A., Palmer, G.H., Ray, S.C., Rikihisa, Y. & Rurangirwa, F.R. (2001). Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and ‘HGE agent’ as subjective synonyms of Ehrlichia phagocytophila . International Journal of Systematic and Evolutionary Microbiology 51, 21452165.
Duron, O., Bouchon, D., Boutin, S., Bellamy, L., Zhou, L., Engelstädter, J. & Hurst, G.D. (2008) The diversity of reproductive parasites among arthropods: Wolbachia do not walk alone. BMC Biology 6, 27.
Espino, C.I., Gómez, T., González, G., do Santos, M.F.B., Solano, J., Sousa, O., Moreno, N., Windsor, D., Ying, A., Vilchez, S. & Osuna, A. (2009) Detection of Wolbachia bacteria in multiple organs and feces of the triatomine insect Rhodnius pallescens (Hemiptera, Reduviidae). Applied and Environmental Microbiology 75, 547550.
Faran, M.E. & Linthicum, K.J. (1981) A handbook of the Amazonian species of Anopheles (Nyssorhynchus) (Diptera: Culcidae). Mosquito System 13, 181.
Fu, Y., Gavotte, L., Mercer, D.R. & Dobson, S.L. (2010) Artificial triple Wolbachia infection in Aedes albopictus yields a new pattern of unidirectional cytoplasmic incompatibility. Applied and Environmental Microbiology 76, 58875891.
Fukatsu, T. & Hosokawa, T. (2002) Capsule-transmitted gut symbiotic bacterium of the Japanese common plataspid stinkbug, Megacopta punctatissima. Applied and Environmental Microbiology 68, 389396.
Galati, E.A.B. (2003) Classificação de Phlebotominae. pp. 23175 in Rangel, E.F. & Lainson, R. (Ed.) Flebotomíneos do Brasil. Rio de Janeiro, Fiocruz.
Gorham, C.H., Fang, Q.Q. & Durden, L.A. (2003) Wolbachia endosymbionts in fleas (Siphonaptera). Journal of Parasitology 89, 283289.
Grimaldi, D. & Engel, M.S. (2005) Evolution of the Insects. p. 772. London, England, Cambridge University Press.
Haegeman, A., Vanholme, B., Jacob, J., Vandekerckhove, T.T.M., Claeys, M., Borgonie, G. & Gheysen, G. (2009) An endosymbiotic bacterium in a plant-parasitic nematode: member of a new Wolbachia supergroup. International Journal for Parasitology 39, 10451054.
Hertig, M. (1936) The rickettsia, Wolbachia pipientis (gen. et sp.n.) and associated inclusions of the mosquito, Culex pipiens . Parasitology 28, 453486.
Hertig, M. & Wolbach, S.B. (1924) Studies on rickettsia-like micro-organisms in insects. Journal of Medical Research 44, 329374.
Hilgenboecker, K., Hammerstein, P., Schlattmann, P., Telschow, A. & Werren, J.H. (2008) How many species are infected with Wolbachia? – a statistical analysis of current data. FEMS microbiology letters 281, 215220.
Hurst, G.D.D. & Jiggins, F.M. (2000) Male-killing bacteria in insects: mechanisms, incidence, and implications. Emerging Infectious Diseases 6, 329336.
Jeyaprakash, A. & Hoy, M.A. (2000) Long PCR improves Wolbachia DNA amplification: wsp sequences found in 76% of sixty-three arthropod species. Insect Molecular Biology 9, 393405.
Kamoda, S., Masui, S., Ishikawa, H. & Sasaki, T. (2000) Wolbachia infection and cytoplasmic incompatibility in the cricket Teleogryllus taiwanemma . Journal of Experimental Biology 203, 25032509.
Kassem, H.A. & Osman, G. (2007) Maternal transmission of Wolbachia in Phlebotomus papatasi (Scopoli). Annals of Tropical Medicine and Parasitology 101, 435440.
Kassem, H.A., Hassan, A.N., Abdel-Hamid, I., Osman, G., El Khalab, E.M. & Madkour, M.A. (2003) Wolbachia infection and the expression of cytoplasmic incompatibility in sandflies (Diptera: Psychodidae) from Egypt. Annals of Tropical Medicine and Parasitology 97, 639644.
Kikuchi, Y. & Fukatsu, T. (2003) Diversity of Wolbachia endosymbionts in heteropteran bugs. Applied and Environmental Microbiology 69, 60826090.
Kittayapong, P., Baisley, K.J., Baimai, V. & O'Neill, S.L. (2000) Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes (Diptera: Culicidae). Journal of Medical Entomology 37, 340345.
Linthicum, K.J. (1988) A revision of the Argyritarsis Section of the subgenus Nyssorhynchus of Anopheles (Diptera: Culicidae). Mosquito System 20, 101271.
Lo, N., Casiraghi, M., Salati, E., Bazzocchi, C. & Bandi, C. (2002). How many Wolbachia supergroups exist? Molecular Biology and Evolution 19, 341346.
Lo, N., Paraskevopoulos, C., Bourtzis, K., O'Neill, S.L., Werren, J.H., Bordenstein, S.R. & Bandi, C. (2007) Taxonomic status of the intracellular bacterium Wolbachia pipientis . International Journal of Systematic and Evolutionary Microbiology 57, 654657.
Martins, C., Souza, R.F. & Bueno, O.C. (2012) Presence and distribution of the endosymbiont Wolbachia among Solenopsis spp. (Hymenoptera: Formicidae) from Brazil and its evolutionary history. Journal of Invertebrate Pathology 109, 287296.
Matsumoto, K., Izri, A., Dumon, H., Raoult, D. & Parola, P. (2008) First detection of Wolbachia spp., including a new genotype, in sand flies collected in Marseille, France. Journal of Medical Entomology 45, 466469.
McMeniman, C.J., Lane, A.M., Fong, A.W., Voronin, D.A., Iturbe-Ormaetxe, I., Yamada, R., McGraw, E.A. & O'Neill, S.L. (2008 Nov) Appl Environ Microbiol. 74(22):6963–9. doi: 10.1128/AEM.01038-08. Epub 2008 Oct 3.
McMeniman, C.J., Lane, R.V., Cass, B.N., Fong, A.W.C., Sidhu, M., Wang, Y-F. & O'Neill, S.L. (2009) Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti . Science 323, 141144.
Morais, S.A., de Almeida, F., Suesdek, L. & Marrelli, M.T. (2012) Low genetic diversity in Wolbachia-infected Culex quinquefasciatus (Diptera: Culicidae) from Brazil and Argentina. Revista do Instituto de Medicina Tropical de São Paulo 54, 325329.
Moreira, L.A., Iturbe-Ormaetxe, I., Jeffery, J.A., Lu, G., Pyke, A.T., Hedges, L.M., Rocha, B.C., Hall-Mendelin, S., Day, A., Riegler, M., Hugo, L.E., Johnson, K.N., Kay, B.H., McGraw, E.A., van den Hurk, A.F., Ryan, P.A. & O'Neill, S.L. (2009) A Wolbachia symbiont in Aedes aegypti limits infection with dengue, Chikungunya, and Plasmodium . Cell 139, 12681278.
Nirgianaki, A., Banks, G.K., Frohlich, D.R., Veneti, Z., Braig, H.R., Miller, T.A., Bedford, I.D., Markham, P.G., Savakis, C. & Bourtzis, K. (2003) Wolbachia infections of the whitefly Bemisia tabaci . Current Microbiology 47, 93101.
O'Neill, S.L., Giordano, R., Colbert, A.M.E., Karr, T.L. & Robertson, H.M. (1992). 16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects. Proceedings of the National Academy of Sciences 89, 26992702.
Ono, M., Braig, H.R., Munstermann, L.E., Ferro, C. & O'Neill, S.L. (2001) Wolbachia infections of phlebotomine sand flies (Diptera: Psychodidae). Journal of Medical Entomology 38, 237241.
Parvizi, P., Fardid, F. & Soleimani, S. (2013) Detection of a new strain of Wolbachia pipientis in Phlebotomus perfiliewi transcaucasicus, a potential vector of visceral Leishmaniasis in north west of Iran, by targeting the major surface protein gene. Journal of Arthropod-Borne Diseases 7, 4655.
Pugedo, H., Barata, R.A., França-Silva, J.C., Silva, J.C. & Dias, E.S. (2005) HP: um modelo aprimorado de armadilha luminosa de sucção para a captura de pequenos insetos. Revista da Sociedade Brasileira de Medicina Tropical 38, 7072.
Rafael, J.A., Melo, G.A.R., de Carvalho, C.J.B., Casari, S.A. & Constantino, R. (2012) Insetos do Brasil: Diversidade e Taxonomia. p. 810. São Paulo, Holos.
Ricci, I., Cancrini, G., Gabrielli, S., D'Amelio, S. & Favia, G. (2002) Searching for Wolbachia (Rickettsiales: Rickettsiaceae) in mosquitoes (Diptera: Culicidae): large polymerase chain reaction survey and new identifications. Journal of Medical Entomology 39, 562567.
Rodriguero, M.S., Confalonieri, V.A., Guedes, J.V. & Lanteri, A.A. (2010) Wolbachia infection in the tribe Naupactini (Coleoptera, Curculionidae): association between thelytokous parthenogenesis and infection status. Insect Molecular Biology 19, 631640.
Rousset, F., Bouchon, D., Pintureau, B., Juchault, P. & Solignac, M. (1992) Wolbachia endosymbionts responsible for various alterations of sexuality in arthropods. Proceedings of the Royal Society B 250, 9198.
Rozen, S. & Skaletsky, H. (2000) Primer3 on the www for general users and for biologist programmers. Methods in Molecular Biology 132, 365386.
Russell, J.A. (2012) The ants (Hymenoptera: Formicidae) are unique and enigmatic hosts of prevalent Wolbachia (Alphaproteobacteria) symbionts. Myrmecological News 16, 723.
Russell, J.A., Funaro, C.F., Giraldo, Y.M., Goldman-Huertas, B., Suh, D., Kronauer, D.J.C., Moreau, C.S. & Pierce, N.E. (2012) A veritable menagerie of heritable bacteria from ants, butterflies, and beyond: broad molecular surveys and a systematic review. PLoS ONE 7, e51027.
Schilthuizen, M. & Stouthamer, R. (1997) Horizontal transmission of parthenogenesis-inducing microbes in Trichogramma wasps. Proceedings of the Royal Society B 264, 361366.
Simões, P.M., Mialdea, G., Reiss, D., Sagot, M-F. & Charlat, S. (2011) Wolbachia detection: an assessment of standard PCR protocols. Molecular Ecology Resources 11, 567572.
Skinner, S.W. (1982) Maternally inherited sex ratio in the parasitoid wasp Nasonia vitripennis . Science 215, 11331134.
Stouthamer, R., Breeuwer, J.A.J. & Hurst, G.D.D. (1999) Wolbachia pipientis: microbial manipulator of arthropod reproduction. Annual Review of Microbiology 53, 71102.
Thipaksorn, A., Jamnongluk, W. & Kittayapong, P. (2003) Molecular evidence of Wolbachia infection in natural populations of tropical odonates. Current microbiology 47, 314318.
Triplehorn, C.A. & Johnson, N.F. (2005) Introduction to the study of insects. p. 809 in Borror and Belmont, Delong's (Ed.). CA, USA, Cengage Learning.
Untergasser, A., Cutcutache, I., Koressaar, T., Ye, J., Faircloth, B.C., Remm, M. & Rozen, S.G. (2012) Primer3 – new capabilities and interfaces. Nucleic Acids Research 40, e115.
Walker, T., Johnson, P.H., Moreira, L.A., Iturbe-Ormaetxe, I., Frentiu, F.D., McMeniman, C.J., Leong, Y.S., Dong, Y., Axford, J., Kriesner, P., Lloyd, A.L., Ritchie, S.A., O'Neill, S.L. & Hoffmann, A.A. (2011) The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations. Nature 2011, 476, 450453.
Weeks, A.R. & Breeuwer, J.A.J. (2001) Wolbachia-induced parthenogenesis in a genus of phytophagous mites. Proceedings of the Royal Society B 268, 22452251.
Wenseleers, T., Ito, F., Van Borm, S., Huybrechts, R., Volckaert, F. & Billen, J. (1998) Widespread occurrence of the micro-organism Wolbachia in ants. Proceedings of the Royal Society B 265, 14471452.
Werren, J.H. (1997) Biology of Wolbachia . Annual Review of Entomology 42, 587609.
Werren, J.H. & Windsor, D.M. (2000) Wolbachia infection frequencies in insects: evidence of a global equilibrium? Proceedings of the Royal Society B 267, 12771285.
Werren, J.H., Zhang, W. & Guo, L.R. (1995a) Evolution and phylogeny of Wolbachia: reproductive parasites of arthropods. Proceedings of the Royal Society B 261, 5563.
Werren, J.H., Windsor, D. & Guo, L. (1995b) Distribution of Wolbachia among neotropical arthropods. Proceedings of the Royal Society B 262, 197204.
Werren, J.H., Baldo, L. & Clark, M.E. (2008) Wolbachia: master manipulators of invertebrate biology. Nature Reviews Microbiology 6, 741751.
West, S.A., Cook, J.M., Werren, J.H. & Godfray, H.C.J. (1998) Wolbachia in two insect host-parasitoid communities. Molecular Ecology 7, 14571465.
Zhou, W., Rousset, F. & O'Neill, S. (1998) Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences. Proceedings of the Royal Society B 265, 509515.
Zug, R. & Hammerstein, P. (2012) Still a host of hosts for Wolbachia: analysis of recent data suggests that 40% of terrestrial arthropod species are infected. PLoS ONE 7, e38544.


Broader prevalence of Wolbachia in insects including potential human disease vectors

  • C.D. de Oliveira (a1), D.S. Gonçalves (a1), L.A. Baton (a1), P.H.F. Shimabukuro (a2), F.D. Carvalho (a1) and L.A. Moreira (a1)...


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