Abstract
The mosquito larval gut is a major consortium for a wide array of microorganisms which has a direct impact on various host traits. The present study focus on the gut bacterial isolation from Aedes aegypti larvae. The phenotypic features of the isolate was elucidated by FE-SEM and identification was done by specific biochemical assay along with 16SrRNA sequencing. The evolutionary significance of bacteria was assessed by MEGA 11.0 software. To evaluate cross gut flora interaction we introduced gut bacteria belonging to common genera like Escherichia Coli, Salmonella typhi, Bacillus subtilis and Staphylococcus aureus to Aliiruegeria sabulilitoris along with broad spectrum antibiotic ampicillin as control in separate culture plates. The ammensalism was observed with Escherichia Coli and Salmonella typhi and stable co-existence with Bacillus subtilis and Staphylococcus aureus. For the first time from India, here we report a single bacterium Aliiruegeria sabulilitoris from Aedes aegypti larval gut. According to MEGA 11.0 software output the bacterium isolated in our study was closely related to samples isolated from neighbouring countries China and South Korea which strongly suggest a common ancestry. Generally larval gut harbour diversegroup of bacteria, here the presence of a single bacterium aggravate our curiosity to investigate future on this phenomenon. The bio-active compound present in the bacterium was found to be Diisobutyl Benzene-1, 2-Dicarboxylate and Tetrahydrodibenzothiophene which revealed by GC–MS technique. Collectively, these findings open a way towards exploiting the composition of mosquito gut flora in predicting the innovative concepts to combat the epidemic threats instigated by deadly diseases transmitted by the Aedes aegypti.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data analysed during the present investigation are available from the corresponding author on reasonable request.
References
Anoopkumar A, Puthur S, Varghese P, Rebello S, Aneesh E (2017) Life cycle, bio-ecology and DNA barcoding of mosquitoes Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse). J Commun Dis 49. https://doi.org/10.24321/0019.5138.201719
Anoopkumar AN, Puthur S, Rebello S, Aneesh EM (2019) Molecular characterization of Aedes, Culex, Anopheles, and Armigeres vector mosquitoes inferred by mitochondrial cytochrome oxidase I gene sequence analysis. Biologia 74(9):1125–1138. https://doi.org/10.2478/s11756-019-00231-0
Boyer S, Sérandour J, Lempérière G, Raveton M, Ravanel P (2006) Do herbicide treatments reduce the sensitivity of mosquito larvae to insecticides? Chemosphere 65(4):721–724. https://doi.org/10.1016/j.chemosphere.2006.02.032
Butt TM, Greenfield BP, Greig C, Maffeis TG, Taylor JW, Piasecka J, Garrido-Jurado I (2013) Metarhizium anisopliae pathogenesis of mosquito larvae: a verdict of accidental death. PLoS One 8(12):e81686. https://doi.org/10.1371/journal.pone.0081686
Cirimotich CM, Ramirez JL, Dimopoulos G (2011) Native microbiota shape insect vector competence for human pathogens. Cell Host Microbe 10(4):307–310. https://doi.org/10.1016/j.chom.2011.09.006
Clarke PH, Cowan S (1952) Biochemical methods for bacteriology. Microbiology 6(1–2):187–197. https://doi.org/10.1099/00221287-6-1-2-187
Claus D (1992) A standardized Gram staining procedure.
Coleman J, Juhn J, James AA (2007) Dissection of midgut and salivary glands from Ae. aegypti mosquitoes. J Vis Exp (5):e228. https://doi.org/10.3791/228
Coon KL, Valzania L, McKinney DA, Vogel KJ, Brown MR, Strand MR (2017) Bacteria-mediated hypoxia functions as a signal for mosquito development. Proc Natl Acad Sci 114(27):E5362–E5369. https://doi.org/10.1073/pnas.1702983114
Cowan ST (1953) Fermentations: biochemical micromethods for bacteriology. Microbiology 8(3): 391–396. https://doi.org/10.1099/00221287-8-3-391
Demaio J, Pumpuni CB, Kent M, Beier JC (1996) The midgut bacterial flora of wild Aedes triseriatus, Culex pipiens, and Psorophora columbiae mosquitoes. Am J Trop Med Hyg 54(2):219–223. https://doi.org/10.4269/ajtmh.1996.54.219
Dhiman RC, Pahwa S, Dhillon G, Dash AP (2010) Climate change and threat of vector-borne diseases in India: are we prepared? Parasitol Res 106(4):763–773. https://doi.org/10.1007/s00436-010-1767-4
Dillon RJ, Dillon V (2004) The gut bacteria of insects: nonpathogenic interactions. Ann Rev Entomol 49(1):71–92
Edwards H (1982) Ion concentration and activity in the haemolymph of Aedes aegypti larvae. J Exp Biol 101(1):143–151. https://doi.org/10.1242/jeb.101.1.143
Hadda TB, Srivastava S, Das B, Salgado-Zamora H, Shaheen U, Bader A, Naseer MM (2014) POM analyses of antimicrobial activity of some 2, 3-armed 4, 5, 6, 7-tetrahydro-1-benzothiophenes: Favourable and unfavourable physico-chemical parameters in design of antibacterial and mycolytic agents. Med Chem Res 23:995–1003. https://doi.org/10.1007/s00044-013-0707-0
Harbach RE (2007) The Culicidae (Diptera): a review of taxonomy, classification and phylogeny. Zootaxa 1668(1):591–638. https://doi.org/10.11646/zootaxa.1668.1.28
Hixson B, Bing XL, Yang X, Bonfini A, Nagy P, Buchon N (2022) A transcriptomic atlas of Aedes aegypti reveals detailed functional organization of major body parts and gut regional specializations in sugar-fed and blood-fed adult females. Elife 11 e76132. https://doi.org/10.7554/eLife.76132
Hougard JM, Duchon S, Zaim M, Guillet P (2002) Bifenthrin: a useful pyrethroid insecticide for treatment of mosquito nets. J Med Entomol 39(3):526–533. https://doi.org/10.1603/0022-2585-39.3.526
Ibrahim H, Uttu AJ, Sallau MS, Iyun ORA (2021) Gas chromatography–mass spectrometry (GC–MS) analysis of ethyl acetate root bark extract of S trychnos innocua (Delile). Beni-Suef Univ J Basic Appl Sci 10:1–8. https://doi.org/10.1186/s43088-021-00156-1
Jayakrishnan L, Sudhikumar AV, Aneesh EM (2018) Role of gut inhabitants on vectorial capacity of mosquitoes. J Vector Borne Dis 55(2):69. https://doi.org/10.4103/0972-9062.242567
Kamboj A, Randhawa H (2012) Pharmacological action and sar of thiophene derivatives: a review. J Pharm Res 5:2676–2682
Kraemer MU, Sinka ME, Duda KA, Mylne A, Shearer FM, Brady OJ, Carvalho RG (2015) The global compendium of Aedes aegypti and Ae. albopictus occurrence. Sci Data 2(1):1–8. https://doi.org/10.1038/sdata.2015.35
Kumar R, Chattopadhyay A, Maitra S, Paul S, Banerjee PK (2013) Isolation and Biochemical characterizations of mid gut microbiota of Culex (Culex quinquefasciatus) mosquitoes in some urban sub urban & rural areas of West Bengal. J Entomol 1:67–78
Laporta GZ, Potter AM, Oliveira JF, Bourke BP, Pecor DB, Linton YM (2023) Global distribution of Aedes aegypti and Aedes albopictus in a Climate Change Scenario of Regional Rivalry. Insects 14(1):49. https://doi.org/10.3390/insects14010049
McMeniman CJ, Lane RV, Cass BN, Fong AW, Sidhu M, Wang Y-F, O’Neill SL (2009) Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti. Science 323(5910):141–144. https://doi.org/10.1126/science.1165326
Min K-T, Benzer S (1997) Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death. Proc Natl Acad Sci 94(20):10792–10796. https://doi.org/10.1073/pnas.94.20.10792
Mougi A (2016) The roles of amensalistic and commensalistic interactions in large ecological network stability. Sci Rep 6(1):1–6. https://doi.org/10.1038/srep29929
Nascimento GG, Locatelli J, Freitas PC, Silva GL (2000) Antibacterial activity of plant extracts and phytochemicals on antibiotic-resistant bacteria. Braz J Microbiol 31:247–256. https://doi.org/10.1590/S1517-83822000000400003
Navon A (2000) Bacillus thuringiensis application in agriculture Entomopathogenic bacteria: from laboratory to field application. Springer, pp. 355–369. https://doi.org/10.1007/978-94-017-1429-7_19
Osei-Poku J, Mbogo C, Palmer W, Jiggins F (2012) Deep sequencing reveals extensive variation in the gut microbiota of wild mosquitoes from Kenya. Mol Ecol 21(20):5138–5150. https://doi.org/10.1111/j.1365-294X.2012.05759.x
Park S, Jung Y-T, Won S-M, Yoon J-H (2014) Pseudoruegeria sabulilitoris sp. nov., isolated from seashore sand. Int J Syst Evol Microbiol 64(Pt_9):3276–3281. https://doi.org/10.1099/ijs.0.066258-0
Pitt T, Barer M (2012) Classification, identification and typing of micro-organisms. Med Microbiol 24. https://doi.org/10.1016/B978-0-7020-4089-4.00018-4
Puthur S, Anoopkumar A, Rebello S, Aneesh EM (2018) Hypericum japonicum: a double-headed sword to combat vector control and cancer. Appl Biochem Biotechnol 186(1):1–11. https://doi.org/10.1007/s12010-018-2713-7
Sarma DK, Kumar M, Dhurve J, Pal N, Sharma P, James MM, Marotta F (2022) Influence of Host Blood Meal Source on Gut Microbiota of Wild Caught Aedes aegypti, a Dominant Arboviral Disease Vector. Microorganisms 10(2):332. https://doi.org/10.3390/microorganisms10020332
Seal M, Chatterjee S (2021) Laboratory studies on ovipositional preferences of malaria vector Anopheles subpictus prevalent in Hooghly district West Bengal India.
Teather RM, Wood PJ (1982) Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl Environ Microbiol 43(4):777–780. https://doi.org/10.1128/aem.43.4.777-780.1982
Valzania L, Martinson VG, Harrison RE, Boyd BM, Coon KL, Brown MR, Strand MR (2018) Both living bacteria and eukaryotes in the mosquito gut promote growth of larvae. PLoS Negl Trop Dis 12(7):e0006638. https://doi.org/10.1371/journal.pntd.0006638
Vijayan A, Rumbo M, Carnoy C, Sirard J-C (2018) Compartmentalized antimicrobial defenses in response to flagellin. Trends Microbiol 26(5):423–435. https://doi.org/10.1016/j.tim.2017.10.008
World Health Organization (2020) Vector borne diseases https://www.who.int. Accessed 02 Mar 2020
Yadav KK, Bora A, Datta S, Chandel K, Gogoi HK, Prasad G, Veer V (2015) Molecular characterization of midgut microbiota of Aedes albopictus and Aedes aegypti from Arunachal Pradesh. India Parasites & Vectors 8(1):1–8. https://doi.org/10.1186/s13071-015-1252-0
Zheng CW, Cheung TMY, Leung GPH (2022) A review of the phytochemical and pharmacological properties of Amauroderma rugosum. Kaohsiung J Med Sci. https://doi.org/10.1002/kjm2.12554
Acknowledgements
The authors would like to thank Department of Zoology, University of Calicut for the laboratory facilities provided.
The authors would like to, thank UGC-SAP for the facilities provided.
Funding
The study was funded by Department of Biotechnology (DBT) (BT/PR39753/FCB/125/95/2020) and Department of Science and Technology(DST)[DST/TDT/SHRI-02/2021(C)].
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics approval and consent to participate
This article does not contain any studies with human participants or animals performed by any of the authors.
Consent for publication
Not required
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Jayakrishnan, L., Aneesh, E.M. Exploiting mosquito microbiome to combat the epidemic threat posed by Aedes aegypti. Int J Trop Insect Sci 43, 805–817 (2023). https://doi.org/10.1007/s42690-023-00990-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42690-023-00990-z