1887

Abstract

Many insects have been associated with actinobacteria in protective symbiosis where antimicrobial metabolites inhibit host pathogens. However, the microbiota of neotropical insects such as the stingless-bee Tetragonisca angustula is poorly explored. T. angustula is a meliponid bee widely distributed in Latin America, its honey is traditionally exploited because of its ethno-pharmacological properties and its antimicrobial activity has been demonstrated. Also, the well-structured nest of this species allows exploration of the microbiota of its different components. Even though Streptomyces spp. have been cultured from stingless-bees, little is known about their role in this insect–microbe relationship. In this study, we examined the association between culturable actinobacteria and T. angustula, and evaluated the isolates’ potential as antimicrobial producers. We isolated 51 actinobacteria from adult bees and different substrates of the hive of T. angustula (pollen and honey storage, garbage pellets and cerumen). We then performed a 16S rRNA phylogenetic analysis that clusters the bacteria to previously described lineages of host-associated Streptomyces . In addition, all the isolates were classified according to their antibacterial activity against human pathogens, measured by a growth inhibition test based on diffusion in agar. More than 50 % of our isolates exhibit antimicrobial activity, mainly to Gram-positive bacteria and fungi and only two against Gram-negative bacteria. Additionally, we obtained electron micrographs of adult bees with what appears to be patches of hyphae with Streptomyces -like cell morphology on their body surface. Our results suggest that T. angustula possibly uptakes and transfers actinobacteria from the environment, acting as vectors for these potentially beneficial organisms. This research provides new insights regarding the microbiota associated with T. angustula and justify future studies exploring the full diversity of the microbial community associated with the hive and the possible exchange of microbes with the crops they pollinate.

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2019-01-24
2024-04-18
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References

  1. Adis J. Thirty million arthropod species – too many or too few?. J Trop Ecol 1990; 6:115–118 [View Article]
    [Google Scholar]
  2. Mora C, Tittensor DP, Adl S, Simpson AG, Worm B. How many species are there on earth and in the ocean?. PLoS Biol 2011; 9:e1001127 [View Article][PubMed]
    [Google Scholar]
  3. Stork NE. How many species of insects and other terrestrial arthropods are there on earth?. Annu Rev Entomol 2018; 63:31–45 [View Article][PubMed]
    [Google Scholar]
  4. Engel MS, Grimaldi DA. New light shed on the oldest insect. Nature 2004; 427:627–630 [View Article][PubMed]
    [Google Scholar]
  5. Chown SL, Convey P. Antarctic entomology. Annu Rev Entomol 2016; 61:119–137 [View Article][PubMed]
    [Google Scholar]
  6. Rosenberg DM, Danks HV, Lehmkuhl DM. Importance of insects in environmental impact assessment. Environmental Management 1986; 10:773–783 [View Article]
    [Google Scholar]
  7. Kaltenpoth M. Actinobacteria as mutualists: general healthcare for insects?. Trends Microbiol 2009; 17:529–535 [View Article][PubMed]
    [Google Scholar]
  8. Douglas AE. Multiorganismal insects: diversity and function of resident microorganisms. Annu Rev Entomol 2015; 60:17–34 [View Article][PubMed]
    [Google Scholar]
  9. Fisher PJ, Stradling DJ, Pegler DN. Leaf cutting ants, their fungus gardens and the formation of basidiomata of Leucoagaricus gongylophorus. Mycologist 1994; 8:128–131 [View Article]
    [Google Scholar]
  10. Brune A. Symbiotic digestion of lignocellulose in termite guts. Nat Rev Microbiol 2014; 12:168–180 [View Article][PubMed]
    [Google Scholar]
  11. van Arnam EB, Currie CR, Clardy J. Defense contracts: molecular protection in insect-microbe symbioses. Chem Soc Rev 2018; 47:1638–1651 [View Article][PubMed]
    [Google Scholar]
  12. Cafaro MJ, Poulsen M, Little AE, Price SL, Gerardo NM et al. Specificity in the symbiotic association between fungus-growing ants and protective Pseudonocardia bacteria. Proc Biol Sci 2011; 278:1814–1822 [View Article][PubMed]
    [Google Scholar]
  13. Kroiss J, Kaltenpoth M, Schneider B, Schwinger MG, Hertweck C et al. Symbiotic streptomycetes provide antibiotic combination prophylaxis for wasp offspring. Nat Chem Biol 2010; 6:261–263 [View Article][PubMed]
    [Google Scholar]
  14. Book AJ, Lewin GR, McDonald BR, Takasuka TE, Wendt-Pienkowski E et al. Evolution of high cellulolytic activity in symbiotic Streptomyces through selection of expanded gene content and coordinated gene expression. PLoS Biol 2016; 14:e1002475 [View Article][PubMed]
    [Google Scholar]
  15. Erwin TL. Tropical forests: their richness in Coleoptera and other arthropod species. The Colleopterist Bulletin 1982; 36:74–75
    [Google Scholar]
  16. Basset Y, Samuelson GA, Allison A, Miller SE. How many species of host-specific insects feed on a species of tropical tree?. Biol J Linn Soc Lond 1996; 59:201–216 [View Article]
    [Google Scholar]
  17. Vargas-Asensio G, Pinto-Tomas A, Rivera B, Hernandez M, Hernandez C et al. Uncovering the cultivable microbial diversity of costa rican beetles and its ability to break down plant cell wall components. PLoS One 2014; 9:e113303 [View Article][PubMed]
    [Google Scholar]
  18. Cheng K, Rong X, Pinto-Tomás AA, Fernández-Villalobos M, Murillo-Cruz C et al. Population genetic analysis of Streptomyces albidoflavus reveals habitat barriers to homologous recombination in the diversification of streptomycetes. Appl Environ Microbiol 2015; 81:966–975 [View Article][PubMed]
    [Google Scholar]
  19. Matarrita-Carranza B, Moreira-Soto RD, Murillo-Cruz C, Mora M, Currie CR et al. Evidence for widespread associations between neotropical hymenopteran insects and actinobacteria. Front Microbiol 2017; 8:8 [View Article][PubMed]
    [Google Scholar]
  20. Michener CD. The Meliponini. In Vit P, Pedro SR, Roubik D. (editors) Pot-Honey, 1st ed. New York: Springer; 2013 pp. 3–17
    [Google Scholar]
  21. van Veen JW, Sommeijer MJ. Colony reproduction in Tetragonisca angustula (Apidae, Meliponini). Insectes Soc 2000; 47:70–75 [View Article]
    [Google Scholar]
  22. Cortopassi-Laurino M, Imperatriz-Fonseca VL, Roubik DW, Dollin A, Heard T et al. Global meliponiculture: challenges and opportunities. Apidologie 2006; 37:275–292 [View Article]
    [Google Scholar]
  23. Zamora G, Arias ML, Aguilar I, Umaña E. Costa Rican pot-honey: its medicinal use and antibacterial effect. In Vit P, Pedro SR, Roubik D. (editors) Pot-Honey, 1st ed. New York: Springer; 2013 pp. 507–512
    [Google Scholar]
  24. Zamora LG, Beukelman CJ, van den Berg AJ, Aerts PC, Quarles van Ufford HC et al. An insight into the antibiofilm properties of Costa Rican stingless bee honeys. J Wound Care 2017; 26:168–177 [View Article][PubMed]
    [Google Scholar]
  25. Jensen PR, Gontang E, Mafnas C, Mincer TJ, Fenical W. Culturable marine actinomycete diversity from tropical Pacific Ocean sediments. Environ Microbiol 2005; 7:1039–1048 [View Article][PubMed]
    [Google Scholar]
  26. Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Syst Bacteriol 1966; 16:313–340 [View Article]
    [Google Scholar]
  27. Hsu SC, Lockwood JL. Powdered chitin agar as a selective medium for enumeration of actinomycetes in water and soil. Appl Microbiol 1975; 29:422–426[PubMed]
    [Google Scholar]
  28. Pridham TG, Anderson P, Foley C, Lindenfelser LA, Hesseltine CW et al. A selection of media for maintenance and taxonomic study of streptomycetes. A selection of media for maintenance and taxonomic study of streptomycetes. Antibiotics Annual 1957947–953
    [Google Scholar]
  29. Chun J, Goodfellow M. A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences. Int J Syst Evol Microbiol 1995; 45:240–245
    [Google Scholar]
  30. Lane DJ. 16S/23S rRNA sequencing. In Goodfellow M, Stackebrandt E. (editors) Nucleic Acid Techniques in Bacterial Systematics New York: John Wiley & Sons; 1991 pp. 115–175
    [Google Scholar]
  31. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990; 215:403–410 [View Article][PubMed]
    [Google Scholar]
  32. Katoh K, Rozewicki J, Yamada KD. MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Brief Bioinform 2017bbx108 [View Article][PubMed]
    [Google Scholar]
  33. Seipke RF, Barke J, Brearley C, Hill L, Yu DW et al. A single Streptomyces symbiont makes multiple antifungals to support the fungus farming ant Acromyrmex octospinosus. PLoS One 2011; 6:e22028 [View Article][PubMed]
    [Google Scholar]
  34. Seipke RF, Barke J, Ruiz-Gonzalez MX, Orivel J, Yu DW et al. Fungus-growing Allomerus ants are associated with antibiotic-producing actinobacteria. Antonie Van Leeuwenhoek 2012; 101:443–447 [View Article][PubMed]
    [Google Scholar]
  35. Poulsen M, Oh DC, Clardy J, Currie CR. Chemical analyses of wasp-associated streptomyces bacteria reveal a prolific potential for natural products discovery. PLoS One 2011; 6:e16763 [View Article][PubMed]
    [Google Scholar]
  36. Madden AA, Grassetti A, Soriano JA, Starks PT. Actinomycetes with antimicrobial activity isolated from paper wasp (Hymenoptera: Vespidae: Polistinae) nests. Environ Entomol 2013; 42:703–710 [View Article][PubMed]
    [Google Scholar]
  37. Kaltenpoth M, Goettler W, Dale C, Stubblefield JW, Herzner G et al. Candidatus Streptomyces philanthi’, an endosymbiotic streptomycete in the antennae of Philanthus digger wasps. Int J Syst Evol Microbiol 2006; 56:1403–1411
    [Google Scholar]
  38. Kaltenpoth M, Yildirim E, Gürbüz MF, Herzner G, Strohm E. Refining the roots of the beewolf-Streptomyces symbiosis: antennal symbionts in the rare genus Philanthinus (Hymenoptera, Crabronidae). Appl Environ Microbiol 2012; 78:822–827 [View Article][PubMed]
    [Google Scholar]
  39. Promnuan Y, Kudo T, Ohkuma M, Chantawannakul P. Streptomyces chiangmaiensis sp. nov. and Streptomyces lannensis sp. nov., isolated from the South-East Asian stingless bee (Tetragonilla collina). Int J Syst Evol Microbiol 2013; 63:1896–1901 [View Article][PubMed]
    [Google Scholar]
  40. Huelsenbeck JP, Ronquist F. MRBAYES: bayesian inference of phylogenetic trees. Bioinformatics 2001; 17:754–755 [View Article][PubMed]
    [Google Scholar]
  41. Tamura K, Peterson D, Peterson N, Stecher G, Nei M et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011; 28:2731–2739 [View Article][PubMed]
    [Google Scholar]
  42. Kim KW. Vapor fixation of intractable fungal cells for simple and versatile scanning electron microscopy. J Phytopathol 2008; 156:125–128 [View Article]
    [Google Scholar]
  43. Kwon Y, Lee J-T, Kim HS, Jeon C, Kwak Y-S. Comparative tomato flower and pollinator hive microbial communities. Journal of Plant Diseases and Protection 2018; 125:115–119 [View Article]
    [Google Scholar]
  44. Corby-Harris V, Maes P, Anderson KE. The bacterial communities associated with honey bee (Apis mellifera) foragers. PLoS One 2014; 9:e95056 [View Article][PubMed]
    [Google Scholar]
  45. Currie CR, Poulsen M, Mendenhall J, Boomsma JJ, Billen J. Coevolved crypts and exocrine glands support mutualistic bacteria in fungus-growing ants. Science 2006; 311:81–83 [View Article][PubMed]
    [Google Scholar]
  46. Scott JJ, Oh DC, Yuceer MC, Klepzig KD, Clardy J et al. Bacterial protection of beetle-fungus mutualism. Science 2008; 322:63 [View Article][PubMed]
    [Google Scholar]
  47. Flórez LV, Scherlach K, Gaube P, Ross C, Sitte E et al. Antibiotic-producing symbionts dynamically transition between plant pathogenicity and insect-defensive mutualism. Nat Commun 2017; 8:15172 [View Article][PubMed]
    [Google Scholar]
  48. Watanabe Y, Shinzato N, Fukatsu T. Isolation of actinomycetes from termites' guts. Biosci Biotechnol Biochem 2003; 67:1797–1801 [View Article][PubMed]
    [Google Scholar]
  49. Patil PB, Zeng Y, Coursey T, Houston P, Miller I et al. Isolation and characterization of a Nocardiopsis sp. from honeybee guts. FEMS Microbiol Lett 2010; 312:110–118 [View Article][PubMed]
    [Google Scholar]
  50. Disayathanoowat T, Young JP, Helgason T, Chantawannakul P. T-RFLP analysis of bacterial communities in the midguts of Apis mellifera and Apis cerana honey bees in Thailand. FEMS Microbiol Ecol 2012; 79:273–281 [View Article][PubMed]
    [Google Scholar]
  51. Jacquemyn H, Lenaerts M, Brys R, Willems K, Honnay O et al. Among-population variation in microbial community structure in the floral nectar of the bee-pollinated forest herb Pulmonaria officinalis L. PLoS One 2013; 8:e56917 [View Article][PubMed]
    [Google Scholar]
  52. Promnuan Y, Kudo T, Chantawannakul P. Actinomycetes isolated from beehives in Thailand. World J Microbiol Biotechnol 2009; 25:1685–1689 [View Article]
    [Google Scholar]
  53. Souza BA, Roubik DW, Barth OM, Heard TA, Enríquez E et al. Composition of stingless bee honey: setting quality standards. Interciencia 2006; 31:
    [Google Scholar]
  54. Rosa CA, Lachance MA. Zygosaccharomyces machadoi sp. n., a yeast species isolated from a nest of the stingless bee Tetragonisca angustula. Lundiana 2005; 6:27–29
    [Google Scholar]
  55. Rosa CA, Lachance MA, Silva JO, Teixeira AC, Marini MM et al. Yeast communities associated with stingless bees. FEMS Yeast Res 2003; 4:271–275 [View Article][PubMed]
    [Google Scholar]
  56. Teixeira AC, Marini MM, Nicoli JR, Antonini Y, Martins RP et al. Starmerella meliponinorum sp. nov., a novel ascomycetous yeast species associated with stingless bees. Int J Syst Evol Microbiol 2003; 53:339–343 [View Article][PubMed]
    [Google Scholar]
  57. Bailey L. Honey Bee Pathology, 2nd ed. London: Academic Press; 1991
    [Google Scholar]
  58. de Lima Procópio RE, da Silva IR, Martins MK, de Azevedo JL, de Araújo JM. Antibiotics produced by Streptomyces. Braz J Infect Dis 2012; 16:466–471 [View Article][PubMed]
    [Google Scholar]
  59. Tiwari K, Gupta RK. Rare actinomycetes: a potential storehouse for novel antibiotics. Crit Rev Biotechnol 2012; 32:108–132 [View Article][PubMed]
    [Google Scholar]
  60. Haeder S, Wirth R, Herz H, Spiteller D. Candicidin-producing Streptomyces support leaf-cutting ants to protect their fungus garden against the pathogenic fungus Escovopsis. Proc Natl Acad Sci USA 2009; 106:4742–4746 [View Article][PubMed]
    [Google Scholar]
  61. Oh DC, Poulsen M, Currie CR, Clardy J. Sceliphrolactam, a polyene macrocyclic lactam from a wasp-associated Streptomyces sp. Org Lett 2011; 13:752–755 [View Article][PubMed]
    [Google Scholar]
  62. Gebhardt K, Schimana J, Krastel P, Dettner K, Rheinheimer J et al. Endophenazines AD, new phenazine antibiotics from the Arthropod associated endosymbiont Streptomyces anulatus. The Journal of Antibiotics 2003; 55:794–800
    [Google Scholar]
  63. Kroiss J, Kaltenpoth M, Schneider B, Schwinger MG, Hertweck C et al. Symbiotic Streptomycetes provide antibiotic combination prophylaxis for wasp offspring. Nat Chem Biol 2010; 6:261–263 [View Article][PubMed]
    [Google Scholar]
  64. Omura S, Ikeda H, Ishikawa J, Hanamoto A, Takahashi C et al. Genome sequence of an industrial microorganism Streptomyces avermitilis: deducing the ability of producing secondary metabolites. Proc Natl Acad Sci USA 2001; 98:12215–12220 [View Article][PubMed]
    [Google Scholar]
  65. Bentley SD, Chater KF, Cerdeño-Tárraga AM, Challis GL, Thomson NR et al. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 2002; 417:141–147 [View Article][PubMed]
    [Google Scholar]
  66. Ikeda H, Ishikawa J, Hanamoto A, Shinose M, Kikuchi H et al. Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat Biotechnol 2003; 21:526–531 [View Article][PubMed]
    [Google Scholar]
  67. van der Heul HU, Bilyk BL, McDowall KJ, Seipke RF, van Wezel GP. Regulation of antibiotic production in Actinobacteria: new perspectives from the post-genomic era. Nat Prod Rep 2018; 35:575–604 [View Article][PubMed]
    [Google Scholar]
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