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Version 2. Wellcome Open Res. 2018 Nov 27 [revised 2018 Nov 27];3:113. doi: 10.12688/wellcomeopenres.14765.2. eCollection 2018.

Novel Wolbachia strains in Anopheles malaria vectors from Sub-Saharan Africa.

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Department of Disease Control, London School of Hygiene & Tropical Medicine, London, WC1E 7HT, UK.
Entomology Branch, Division of Parasitic Diseases and Malaria, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, 30033, USA.
Department of Pharmacology and Toxicology, University of Texas Medical Branch at Galveston, Galveston, Texas, USA.
Laboratoire d'entomologie médicale et parasitologie, Centre de Recherche en Sciences Naturelles (CRSN/LWIRO), Sud-Kivu, Congo, Democratic Republic.
Unité d'Entomologie Médicale, Institut Pasteur de Madagascar, Antananarivo, Madagascar.
Nationale de Lutte contre le Paludisme, Ministere de la Sante, Conakry, Guinea.
National Institute of Biomedical Research, Kinshasa, Congo, Democratic Republic.
Department of Medical Microbiology, University of Ghana, Accra, Ghana.
Malaria Consortium, London, EC2A 4LT, UK.
Department of Pathology, University of Texas Medical Branch, Galveston, Texas, USA.
The US President's Malaria Initiative and Entomology Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, 30329-4027, USA.
Department of Pathology, Institute for Human Infections and Immunity, Center for Tropical Diseases, Center for Biodefense and Emerging Infectious Disease, University of Texas Medical Branch, Galveston, Texas, USA.


Background:  Wolbachia, a common insect endosymbiotic bacterium that can influence pathogen transmission and manipulate host reproduction, has historically been considered absent from the  Anopheles (An.) genera, but has recently been found in  An. gambiae s.l. populations in West Africa.  As there are numerous  Anopheles species that have the capacity to transmit malaria, we analysed a range of species across five malaria endemic countries to determine  Wolbachia prevalence rates, characterise novel  Wolbachia strains and determine any correlation between the presence of  PlasmodiumWolbachia and the competing bacterium  Asaia. Methods:  Anopheles adult mosquitoes were collected from five malaria-endemic countries: Guinea, Democratic Republic of the Congo (DRC), Ghana, Uganda and Madagascar, between 2013 and 2017.  Molecular analysis was undertaken using quantitative PCR, Sanger sequencing,  Wolbachia multilocus sequence typing (MLST) and high-throughput amplicon sequencing of the bacterial  16S rRNA gene.  Results: Novel  Wolbachia strains were discovered in five species:  An. coluzziiAn. gambiae s.s.,  An. arabiensisAn. moucheti and  An. species A, increasing the number of  Anopheles species known to be naturally infected. Variable prevalence rates in different locations were observed and novel strains were phylogenetically diverse, clustering with  Wolbachia supergroup B strains.  We also provide evidence for resident strain variants within  An. species A.  Wolbachia is the dominant member of the microbiome in  An. moucheti and  An. species A but present at lower densities in  An. coluzzii.  Interestingly, no evidence of  Wolbachia/Asaia co-infections was seen and  Asaia infection densities were shown to be variable and location dependent.  Conclusions: The important discovery of novel  Wolbachia strains in  Anopheles provides greater insight into the prevalence of resident  Wolbachia strains in diverse malaria vectors.  Novel  Wolbachia strains (particularly high-density strains) are ideal candidate strains for transinfection to create stable infections in other  Anopheles mosquito species, which could be used for population replacement or suppression control strategies.


Anopheles; Asaia; Wolbachia; endosymbionts; malaria; mosquitoes

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