Carryover effects of larval exposure to different environmental bacteria drive adult trait variation in a mosquito vector

The adult phenotype of mosquitoes depends on the types of bacteria encountered environmentally during development. Conditions experienced during larval development of holometabolous insects can affect adult traits, but whether differences in the bacterial communities of larval development sites contribute to variation in the ability of insect vectors to transmit human pathogens is unknown. We addressed this question in the mosquito Aedes aegypti, a major arbovirus vector breeding in both sylvatic and domestic habitats in Sub-Saharan Africa. Targeted metagenomics revealed differing bacterial communities in the water of natural breeding sites in Gabon. Experimental exposure to different native bacterial isolates during larval development resulted in significant differences in pupation rate and adult body size but not life span. Larval exposure to an Enterobacteriaceae isolate resulted in decreased antibacterial activity in adult hemolymph and reduced dengue virus dissemination titer. Together, these data provide the proof of concept that larval exposure to different bacteria can drive variation in adult traits underlying vectorial capacity. Our study establishes a functional link between larval ecology, environmental microbes, and adult phenotypic variation in a holometabolous insect vector.

[1]  K. Ng’habi,et al.  Effect of larval crowding on mating competitiveness of Anopheles gambiae mosquitoes , 2005, Malaria Journal.

[2]  Mark R. Brown,et al.  Mosquitoes rely on their gut microbiota for development , 2014, Molecular ecology.

[3]  C. Mbogo,et al.  Deep sequencing reveals extensive variation in the gut microbiota of wild mosquitoes from Kenya , 2012, Molecular ecology.

[4]  W. Takken,et al.  Larval nutrition differentially affects adult fitness and Plasmodium development in the malaria vectors Anopheles gambiae and Anopheles stephensi , 2013, Parasites & Vectors.

[5]  R. Jarman,et al.  Genetic Mapping of Specific Interactions between Aedes aegypti Mosquitoes and Dengue Viruses , 2013, PLoS genetics.

[6]  H. Overgaard,et al.  Comparative assessment of the bacterial communities associated with Aedes aegypti larvae and water from domestic water storage containers , 2014, Parasites & Vectors.

[7]  D. Marshall,et al.  FITNESS CONSEQUENCES OF LARVAL TRAITS PERSIST ACROSS THE METAMORPHIC BOUNDARY , 2011, Evolution; international journal of organic evolution.

[8]  A. Douglas.  Multiorganismal insects: diversity and function of resident microorganisms. , 2015, Annual review of entomology.

[9]  A. Fontaine,et al.  Excretion of dengue virus RNA by Aedes aegypti allows non-destructive monitoring of viral dissemination in individual mosquitoes , 2016, Scientific Reports.

[10]  D. Yee,et al.  Constitutive Differences between Natural and Artificial Container Mosquito Habitats: Vector Communities, Resources, Microorganisms, and Habitat Parameters , 2012, Journal of medical entomology.

[11]  J. Pascale,et al.  Reciprocal Tripartite Interactions between the Aedes aegypti Midgut Microbiota, Innate Immune System and Dengue Virus Influences Vector Competence , 2012, PLoS neglected tropical diseases.

[12]  N. Moran.  Adaptation and Constraint in the Complex Life Cycles of Animals , 1994 .

[13]  L. P. Lounibos,et al.  LARVAL COMPETITION DIFFERENTIALLY AFFECTS ARBOVIRUS INFECTION IN AEDES MOSQUITOES. , 2005, Ecology.

[14]  Dominique Lavenier,et al.  Multiple comparative metagenomics using multiset k-mer counting , 2016, PeerJ Comput. Sci..

[15]  J. Arredondo‐Jimenez,et al.  Cyanobacteria Associated with Anopheles albimanus (Diptera: Culicidae) Larval Habitats in Southern Mexico , 2002, Journal of medical entomology.

[16]  J. Ramirez,et al.  Native microbiota shape insect vector competence for human pathogens. , 2011, Cell host & microbe.

[17]  Jacques Ravel,et al.  An improved dual-indexing approach for multiplexed 16S rRNA gene sequencing on the Illumina MiSeq platform , 2014, Microbiome.

[18]  L. P. Lounibos,et al.  Larval environmental temperature and the susceptibility of Aedes albopictus Skuse (Diptera: Culicidae) to Chikungunya virus. , 2010, Vector borne and zoonotic diseases.

[19]  H. Shahbazkia,et al.  Midgut Microbiota of the Malaria Mosquito Vector Anopheles gambiae and Interactions with Plasmodium falciparum Infection , 2012, PLoS pathogens.

[20]  E. Muturi,et al.  Temperature and Density-Dependent Effects of Larval Environment on Aedes aegypti Competence for an Alphavirus , 2012, Journal of vector ecology : journal of the Society for Vector Ecology.

[21]  J. Powell,et al.  History of domestication and spread of Aedes aegypti - A Review , 2013, Memorias do Instituto Oswaldo Cruz.

[22]  L. P. Lounibos,et al.  Larval competition alters susceptibility of adult Aedes mosquitoes to dengue infection , 2008, Proceedings of the Royal Society B: Biological Sciences.

[23]  C. Valiente Moro,et al.  French invasive Asian tiger mosquito populations harbor reduced bacterial microbiota and genetic diversity compared to Vietnamese autochthonous relatives , 2015, Front. Microbiol..

[24]  M. Mogi,et al.  Biology of mosquitoes. , 1987 .

[25]  E. McGraw,et al.  Wolbachia pipientis: an expanding bag of tricks to explore for disease control. , 2010, Trends in parasitology.

[26]  R. Bhatnagar,et al.  Bacterial diversity analysis of larvae and adult midgut microflora using culture-dependent and culture-independent methods in lab-reared and field-collected Anopheles stephensi-an Asian malarial vector , 2009, BMC Microbiology.

[27]  S. Kennedy,et al.  Bacteriocin from epidemic Listeria strains alters the host intestinal microbiota to favor infection , 2016, Proceedings of the National Academy of Sciences.

[28]  B. Alto,et al.  Temperature and dengue virus infection in mosquitoes: independent effects on the immature and adult stages. , 2013, The American journal of tropical medicine and hygiene.

[29]  C. Valiente Moro,et al.  Diversity of culturable bacteria including Pantoea in wild mosquito Aedes albopictus , 2013, BMC Microbiology.

[30]  Ying Wang,et al.  Dynamic Gut Microbiome across Life History of the Malaria Mosquito Anopheles gambiae in Kenya , 2011, PloS one.

[31]  C. Murdock,et al.  Capacity of mosquitoes to transmit malaria depends on larval environment , 2014, Parasites & Vectors.

[32]  W. Takken,et al.  Effect of rearing temperature and larval density on larval survival, age at pupation and adult size of Anopheles gambiae , 1992 .

[33]  L. P. Lounibos,et al.  Size alters susceptibility of vectors to dengue virus infection and dissemination. , 2008, The American journal of tropical medicine and hygiene.

[34]  Zhiyong Xi,et al.  The Endosymbiotic Bacterium Wolbachia Induces Resistance to Dengue Virus in Aedes aegypti , 2010, PLoS pathogens.

[35]  C. Schal,et al.  Diversity of Bacterial Communities in Container Habitats of Mosquitoes , 2008, Microbial Ecology.

[36]  A. Telang,et al.  Larval nutritional stress affects vector immune traits in adult yellow fever mosquito Aedes aegypti (Stegomyia aegypti) , 2012, Medical and veterinary entomology.

[37]  P. Mavingui,et al.  Diversity and function of bacterial microbiota in the mosquito holobiont , 2013, Parasites & Vectors.

[38]  R. Rippka,et al.  Selection of cyanobacteria isolated from mosquito breeding sites as a potential food source for mosquito larvae , 1991, Applied and environmental microbiology.

[39]  E. Walker,et al.  NUTRIENT DYNAMICS, BACTERIAL POPULATIONS, AND MOSQUITO PRODUCTIVITY IN TREE HOLE ECOSYSTEMS AND MICROCOSMS' , 1991 .

[40]  T. Scott,et al.  Dengue-1 Virus Clade Replacement in Thailand Associated with Enhanced Mosquito Transmission , 2011, Journal of Virology.

[41]  A. Jacot,et al.  Juvenile immune system activation induces a costly upregulation of adult immunity in field crickets Gryllus campestris , 2005, Proceedings of the Royal Society B: Biological Sciences.

[42]  K. Lerdthusnee,et al.  Meconial Peritrophic Membranes and the Fate of Midgut Bacteria During Mosquito (Diptera: Culicidae) Metamorphosis , 2001, Journal of medical entomology.

[43]  L. Kramer,et al.  Dissecting vectorial capacity for mosquito-borne viruses. , 2015, Current opinion in virology.

[44]  L. Kramer,et al.  Global genetic diversity of Aedes aegypti , 2016, Molecular ecology.

[45]  M. Strand,et al.  Insect hemocytes and their role in immunity. , 2002, Insect biochemistry and molecular biology.

[46]  D. Daffonchio,et al.  Delayed larval development in Anopheles mosquitoes deprived of Asaiabacterial symbionts , 2012, BMC Microbiology.

[47]  G. Favia,et al.  Identification of the Midgut Microbiota of An. stephensi and An. maculipennis for Their Application as a Paratransgenic Tool against Malaria , 2011, PloS one.

[48]  K. P. Sauer,et al.  Gender differences and individual variation in the immune system of the scorpionfly Panorpa vulgaris (Insecta: Mecoptera). , 2000, Developmental and comparative immunology.

[49]  G. Dimopoulos,et al.  Chromobacterium Csp_P Reduces Malaria and Dengue Infection in Vector Mosquitoes and Has Entomopathogenic and In Vitro Anti-pathogen Activities , 2014, PLoS pathogens.

[50]  G. Hughes,et al.  The microbiome modulates arbovirus transmission in mosquitoes. , 2015, Current opinion in virology.

[51]  J. Githure,et al.  Influence of soil quality in the larval habitat on development of Anopheles gambiae Giles , 2006, Journal of vector ecology : journal of the Society for Vector Ecology.

[52]  R. Stoks,et al.  FITNESS EFFECTS FROM EGG TO REPRODUCTION: BRIDGING THE LIFE HISTORY TRANSITION , 2005 .

[53]  T. Lefèvre,et al.  Evidence for carry-over effects of predator exposure on pathogen transmission potential , 2015, Proceedings of the Royal Society B: Biological Sciences.

[54]  J. Beier,et al.  Larval habitats of Anopheles gambiae s.s. (Diptera: Culicidae) influences vector competence to Plasmodium falciparum parasites , 2007, Malaria Journal.

[55]  Bodil N. Cass,et al.  Stable Introduction of a Life-Shortening Wolbachia Infection into the Mosquito Aedes aegypti , 2009, Science.

[56]  S. Ritchie,et al.  Dynamics of the “Popcorn” Wolbachia Infection in Outbred Aedes aegypti Informs Prospects for Mosquito Vector Control , 2011, Genetics.

[57]  K. Lerdthusnee,et al.  Meconial peritrophic matrix structure, formation, and meconial degeneration in mosquito pupae/pharate adults: histological and ultrastructural aspects. , 2005, Journal of medical entomology.

[58]  D. Daffonchio,et al.  Mosquito-Bacteria Symbiosis: The Case of Anopheles gambiae and Asaia , 2010, Microbial Ecology.

[59]  W. Takken,et al.  Competitive interactions between larvae of the malaria mosquitoes Anopheles arabiensis and Anopheles gambiae under semi-field conditions in western Kenya. , 2009, Acta tropica.

[60]  C. Valiente Moro,et al.  Pyrosequencing 16S rRNA genes of bacteria associated with wild tiger mosquito Aedes albopictus: a pilot study , 2014, Front. Cell. Infect. Microbiol..

[61]  C. Barillas-Mury,et al.  Hemocyte Differentiation Mediates Innate Immune Memory in Anopheles gambiae Mosquitoes , 2010, Science.

[62]  A. Enayati,et al.  dentification of bacterial microflora in the midgut of the larvae and adult of wild aught Anopheles stephensi : A step toward finding suitable paratransgenesis andidates , 2011 .

[63]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[64]  M. Riehle,et al.  The impact of larval and adult dietary restriction on lifespan, reproduction and growth in the mosquito Aedes aegypti , 2010, Experimental Gerontology.

[65]  P. Mavingui,et al.  Bacterial diversity of field-caught mosquitoes, Aedes albopictus and Aedes aegypti, from different geographic regions of Madagascar. , 2011, FEMS microbiology ecology.

[66]  R. Christen,et al.  Composition of Anopheles coluzzii and Anopheles gambiae microbiota from larval to adult stages. , 2014, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[67]  P. Kittayapong,et al.  Maternal transmission efficiency of Wolbachia superinfections in Aedes albopictus populations in Thailand. , 2002, The American journal of tropical medicine and hygiene.

[68]  Mark R. Brown,et al.  Mosquitoes host communities of bacteria that are essential for development but vary greatly between local habitats , 2016, Molecular ecology.