Developmental succession of the microbiome of Culex mosquitoes

BMC Microbiology

William E. Walton²  Josh D. Neufeld³  Olle Terenius¹  Richard Stouthamer²  Paul Rugman-Jones²  Michael W. Hall³  Dagne Duguma⁴ 

[1] Department of Ecology, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden;Department of Entomology, University of California Riverside, Riverside 92521, CA, USA;Department of Biology, University of Waterloo, Waterloo N2L 3G1, Ontario, Canada;Present address: Florida Medical Entomology Laboratory, University of Florida, Vero Beach 32962, FL, USA

关键词: Transstadial transmission;    Biopesticide;    Bacteria;    Outdoor mesocosms;    Thorsellia;   

Others  :  1227622
DOI  :  10.1186/s12866-015-0475-8

 received in 2015-01-21, accepted in 2015-06-30,  发布年份 2015

【 摘 要 】

Background

The native microflora associated with mosquitoes have important roles in mosquito development and vector competence. Sequencing of bacterial V3 region from 16S rRNA genes across the developmental stages of Culex mosquitoes (early and late larval instars, pupae and adults) was used to test the hypothesis that bacteria found in the larval stage of Culex are transstadially transmitted to the adult stage, and to compare the microbiomes of field-collected versus laboratory-reared mosquitoes.

Results

Beta diversity analysis revealed that bacterial community structure differed among three life stages (larvae, pupae and adults) of Culex tarsalis. Although only ~2 % of the total number of bacterial OTUs were found in all stages, sequences from these OTUs accounted for nearly 82 % of the total bacterial sequences recovered from all stages. Thorsellia (Gammaproteobacteria) was the most abundant bacterial taxon found across all developmental stages of field-collected Culex mosquitoes, but was rare in mosquitoes from laboratory-reared colonies. The proportion of Thorsellia sequences in the microbiomes of mosquito life stages varied ontogenetically with the greatest proportions recovered from the pupae of C. tarsalis and the lowest from newly emerged adults. The microbiome of field-collected late instar larvae was not influenced significantly by differences in the microbiota of the habitat due to habitat age or biopesticide treatments. The microbiome diversity was the greatest in the early instar larvae and the lowest in laboratory-reared mosquitoes.

Conclusions

Bacterial communities in early instar C. tarsalis larvae were significantly more diverse when compared to late instar larvae, pupae and newly emerged adults. Some of the bacterial OTUs found in the early instar larvae were also found across developmental stages. Thorsellia dominated the bacterial communities in field-collected immature stages but occurred at much lower relative abundance in adults. Differences in microbiota observed in larval habitats did not influence bacterial community profiles of late instar larvae or adults. However, bacterial communities in laboratory-reared C. tarsalis larvae differed significantly from the field. Determining the role of Thorsellia in mosquitoes and its distribution across different species of mosquitoes warrants further investigation.

【 授权许可】

   
2015 Duguma et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

Fig. 7.

Fig. 8.

Fig. 9.

【 参考文献 】

• [1]Merritt RW, Dadd RH, Walker ED. Feeding behavior, natural food, and nutritional relationships of larval mosquitoes. Annu Rev Entomol. 1992; 37:349-374.
• [2]Clements AN. The biology of mosquitoes: Development, nutrition and reproduction. Chapman & Hall, London; 1992.
• [3]Minard G, Mavingui P, Moro CV. Diversity and function of bacterial microbiota in the mosquito holobiont. Parasit Vectors. 2013; 6:1-12. BioMed Central Full Text
• [4]Coon KL, Vogel KJ, Brown MR, Strand MR. Mosquitoes rely on their gut microbiota for development. Mol Ecol. 2014; 23:2727-2739.
• [5]Pumpuni CB, Beier MS, Nataro JP, Guers LD, Davis JR. Plasmodium falciparum: inhibition of sporogonic development in Anopheles stephensi by Gram-negative Bacteria. Exp Parasitol. 1993; 7:195-199.
• [6]Okech B, Gouagna L, Yan G, Githure J, Beier J. Larval habitats of Anopheles gambiae s.s. (Diptera: Culicidae) influences vector competence to Plasmodium falciparum parasites. Malar J. 2007; 6:50. BioMed Central Full Text
• [7]Cirimotich CM, Ramirez JL, Dimopoulos G. Native microbiota shape insect vector competence for human pathogens. Cell Host Microb. 2011; 10:307-310.
• [8]Takken W, Smallegange RC, Vigneau AJ, Johnston V, Brown M, Mordue-Luntz AJ, Billingsley PF. Larval nutrition differentially affects adult fitness and Plasmodium development in the malaria vectors Anopheles gambiae and Anopheles stephensi. Parasit Vectors. 2013; 6:345. BioMed Central Full Text
• [9]Dong Y, Manfredini F, Dimopoulos G. Implication of the mosquito midgut microbiota in the defense against malaria parasites. PLoS Patho. 2009; 5: Article ID e1000423
• [10]Ricci I, Valzano M, Ulissi U, Epis S, Cappelli A, Favia G. Symbiotic control of mosquito borne disease. Patho Global Health. 2012; 106:380-385.
• [11]Bando H, Okado K, Guelbeogo W, Badolo A, Aonuma H, et al. Intra-specific diversity of Serratia marcescens in Anopheles mosquito midgut defines Plasmodium transmission capacity. Sci Rep. 2013;3.
• [12]Riehle MA, Jacobs-Lorena M. Using bacteria to express and display anti-parasite molecules in mosquitoes: current and future strategies. Insect Biochem Mol Biol. 2005; 35:699-707.
• [13]Boissière A, Tchioffo MT, Bachar D, Abate L, Marie A et al.. Midgut microbiota of the malaria mosquito vector Anopheles gambiae and interactions with Plasmodium falciparum infection. PLoS Patho. 2012; 8: Article ID e1002742
• [14]Osei-Poku J, Mbogo C, Palmer W, Jiggins F. Deep sequencing reveals extensive variation in the gut microbiota of wild mosquitoes from Kenya. Mol Ecol. 2012; 21:5138-5150.
• [15]Wang S, Ghosh AK, Bongio N, Stebbings KA, Lampe D, Jacobs-Lorena M. Fighting malaria with engineered symbiotic bacteria from vector mosquitoes. Proc Natl Acad Sci USA. 2012; 109:12734-12739.
• [16]Caljon G, De Vooght L, Van den Abbeele J. Options for the delivery of anti-pathogen molecules in arthropod vectors. J Invert Pathol. 2013; 112:S75-S82.
• [17]Moll RM, Romoser WS, Modrakowski MC, Moncayo AC, Lerdthusnee K. Meconial peritrophic membranes and the fate of midgut bacteria during mosquito (Diptera: Culicidae) metamorphosis. J Med Entomol. 2001; 38:29-32.
• [18]Rani A, Sharma A, Rajagopal R, Adak T, Bhatnagar R. 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. BMC Microbiol. 2009; 9:96. BioMed Central Full Text
• [19]Wang Y, Gilbreath T, Kukutla P, Yan G, Xu J. Dynamic gut microbiome across life history of the malaria mosquito Anopheles gambiae in Kenya. PLoS ONE. 2011; 6: Article ID e24767
• [20]Chavshin AR, Oshaghi MA, Vatandoost H, Yakhchali B, Raeisi A, Zarenejad F. Escherichia coli expressing a green fluorescent protein (GFP) in Anopheles stephensi: a preliminary model for paratransgenesis. Symbiosis. 2013; 60:17-24.
• [21]Chavshin AR, Oshaghi MA, Vatandoost H, Yakhchali B, Raeisi A, Zarenejad F, Terenius O. Malpighian tubules are important determinants of Pseudomonas transstadial transmission and longtime persistence in Anopheles stephensi. Parasit Vectors. 2015; 8:36. BioMed Central Full Text
• [22]Chandler JA, Lang JM, Bhatnagar S, Eisen JA, Kop A. Bacterial communities of diverse Drosophila species: ecological context of a host–microbe model system. PLoS Genet. 2011; 7: Article ID e1002272
• [23]Duguma D, Rugman-Jones P, Kaufman MG, Hall MW, Neufeld JD, Stouthamer R, Walton WE. Bacterial communities associated with Culex mosquito larvae and two emergent aquatic plants of bioremediation importance. PLoS One. 2013; 8: Article ID e72522
• [24]Lindh JM, Terenius O, Faye I. 16S rRNA gene-based identification of midgut bacteria from field-caught Anopheles gambiae sensu lato and A. funestus mosquitoes reveals new species related to known insect symbionts. Appl Environ Microbiol. 2005; 71:7217-7223.
• [25]Kämpfer P, Lindh J, Terenius O, Haghdoost S, Falsen E, Busse HJ, Faye I. Thorsellia anophelis gen nov, sp nov, a new member of the Gammaproteobacteria. Int J Syst Evol Microbiol. 2006; 56:335-338.
• [26]Kämpfer P, Glaeser SP, Nilsson LK, Eberhard T, Håkansson S, Guy L, Roos S, Busse H, Terenius O. Proposal of Thorsellia kenyensis sp. nov. and Thorsellia kandunguensis sp. nov., isolated from the larvae of Anopheles arabiensis as members of the family Thorselliaceae fam. nov. Int J Syst Evol Microbiol. 2014; 65:444-451.
• [27]Briones A, Shililu J, Githure J, Novak R, Raskin L. Thorsellia anophelis is the dominant bacterium in a Kenyan population of adult Anopheles gambiae mosquitoes. The ISME J. 2008; 2:74-82.
• [28]Chavshin AR, Oshaghi MA, Vatandoost H, Pourmand MR, Raeisi A, Terenius O. Isolation and identification of culturable bacteria from wild Anopheles culicifacies, a first step in a paratransgenesis approach. Parasit Vector. 2014; 7:419. BioMed Central Full Text
• [29]Muturi EJ, Orindi BO, Kim CH. Effect of leaf type and pesticide exposure on abundance of bacterial taxa in mosquito larval habitats. PLoS One. 2013; 8: Article ID e71812
• [30]Kaufman MG, Chen S, Walker ED. Leaf-associated bacterial and fungal taxa shifts in response to larvae of the tree hole mosquito, Ochlerotatus triseriatus. Microb Ecol. 2008; 55:673-684.
• [31]Xu Y, Chen S, Kaufman MG, Maknojia S, Bagdsarian M, Walker ED. Bacterial community structure in treehole habitats of Ochlerotatus triseiatus: influences of larval feeding. J Am Mosq Control Assoc. 2008; 24:219-227.
• [32]Walker ED, Kaufman MG, Merritt RW. An acute trophic cascade among microorganisms in the tree hole ecosystem following removal of omnivorous mosquito larvae. Community Ecol. 2010; 11:171-178.
• [33]Duguma D, Hall MW, Rugman-Jones P, Neufeld JD, Stouthamer R, Walton WE. Microbial communities and nutrient dynamics in experimental microcosms are altered after application of a high dose of Bti. J Appl Ecol. 2015; 52:763-773.
• [34]Nkya TE, Akhouayri I, Kisinza W, David JP. Impact of environment on mosquito response to pyrethroid insecticides: facts, evidences and prospects. Insect Biochem Mol Biol. 2013; 43:407-416.
• [35]Xia X, Zheng D, Zhong H, Qin B, Gurr GM et al.. DNA sequencing reveals the midgut microbiota of diamondback moth, Plutella xylostella (L.) and a possible relationship with insecticide resistance. PLoS One. 2013; 8:e68852.
• [36]Rasgon J, Scott T. An initial survey for Wolbachia (Rickettsiales: Rickettsiaceae) infections in selected California mosquitoes (Diptera: Culicidae). J Med Entomol. 2004; 41:255-257.
• [37]Peck GW, Walton WE. Effects of bacterial food quality and density on growth and whole body stoichiometry of Culex quinquefasciatus and Culex tarsalis (Diptera: Culicidae). J Med Entomol. 2006; 43:25-33.
• [38]Nilsson L: Isolation of Thorsellia from Kenyan Anopheles gambiae sensu lato and their breeding waters. MS thesis. Swedish University of Agricultural Sciences; 2012
• [39]Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 2013; 30:2725-2729.
• [40]Tamura K, Nei M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol. 1993; 10:512-526.
• [41]Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK et al.. QIIME allows analysis of high-throughput community sequencing data. Nat Methods. 2010; 7:335-336.
• [42]Lynch MD, Masella AP, Hall MW, Bartram AK, Neufeld JD. AXIOME: automated exploration of microbial diversity. GigaScience. 2013; 2:3. BioMed Central Full Text
• [43]Li W, Godzik A. Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics. 2006; 22:1658-1659.
• [44]Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Glöckner FO. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucl Acids Res. 2013; 41:D590-D596.
• [45]Mielke PW, Berry KJ, Johnson ES. Multi-response permutation procedures for a priori classifications. Commun Stat Theor Methods. 1976; 5:1409-1424.