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Wellcome Open Res. 2018 Sep 12;3:113. doi: 10.12688/wellcomeopenres.14765.1. 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.  As there are numerous Anopheles species that have the capacity to transmit malaria, we analysed a range of species to determine Wolbachia prevalence rates, characterise novel Wolbachia strains and determine any correlation between the presence of Plasmodium, Wolbachia and the competing endosymbiotic 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 of samples 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. coluzzii, An. gambiae s.s., An. arabiensis, An. 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 also 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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