Unveiling the Cultivation-Dependent Gut Bacterial Microbiota in the Feral Stingless Bee, Tetragonula iridipennis Smith (hymenoptera: Apidae)

Authors

  • B. Saai Vignesh Department of Agricultural Entomology, Tamil Nadu Agricultural University, Madurai, Tamil Nadu, India
  • J. Jayaraj Agricultural College and Research Institute, Tamil Nadu Agricultural University, Chettinad, Tamil Nadu, India
  • R. Nalini Department of Agricultural Entomology, Tamil Nadu Agricultural University, Madurai, Tamil Nadu, India
  • K. Kumutha Department of Agricultural Microbiology, Tamil Nadu Agricultural University, Madurai, Tamil Nadu, India
  • M. R. Srinivasan Department of Agricultural Entomology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India
  • M. Jayakanthan Department of Bioinformatics, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India.
  • K. Suresh Department of Agricultural Entomology, ICAR-KVK, Tamil Nadu Agricultural University, Madurai, Tamil Nadu, India
  • P. Sabatina Department of Agricultural Entomology, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India

DOI:

https://doi.org/10.13102/sociobiology.v72i3.11205

Keywords:

16S rRNA gene sequencing, Bacillus sp., Clostridium sp., Citrobacter sp., Social insect, stingless bee gut bacteria

Abstract

The feral stingless bee, Tetragonula iridipennis, is a vital pollinator in diverse ecosystems and produces valuable honey with unique characteristics. This study aimed to explore the gut microbiota of T. iridipennis by identifying and characterizing its bacterial communities and examining the interactions between gut bacteria and pathogens. Total Plate counts (TPC) were assessed after a day of incubation on nutrient agar at 37 °C, revealing the highest TPC from SL1 (7.00 ± 0.03 log cfu/ml) and the lowest from SL3 (6.79 ± 0.04 log cfu/ml). Eleven bacterial isolates were recovered and classified through morphological characterization and Gram staining. All bacterial isolates were subject to antimicrobial assessment against infective bacteria. The isolates SB1, SB4, SBT2, and SBM1 showed better antagonistic effect (12.67 ± 2.08; 11.33 ± 1.53; 10.67 ± 4.16 and 9.67 ± 1.53 mm) among the eleven isolates against the pathogen Staphylococcus aureus, respectively. These selected isolates were taken into molecular characterization through 16S rRNA gene sequencing. The amplified 1500 base pair fragments showed more than 98% similarity with known sequences in the GenBank database. The isolates were identified as Bacillus subtilis (SB1), Bacillus tropicus (SB4), Clostridium tunisiense (SBT2), and Citrobacter freundii (SBM1). The phylogenetic analysis using MEGA 11 and BLASTn revealed that B. subtilis SB(CM)-1 had 77% similarity with B. subtilis C785F of South Korea, B. tropicus SB(OD)-1 had 69% similarity with B. tropicus SA39 from Pakistan, C. tunisiense SB(TK)-2 had 100% similarity with C. tunisiense from Japan, and Citrobacter freundii SCF(CM)-2 had 100% similarity with Citrobacter sp. from Brazil.

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References

Anderson, K.E., Sheehan, T.H., Mott, B.M., Maes, P., Snyder, L., Schwan, M.R., Walton, A., Jones, B.M. & Corby-Harris, V. (2013). Microbial ecology of the hive and pollination landscape: bacterial associates from floral nectar, the alimentary tract and stored food of honey bees (Apis mellifera). PLoS ONE, 8: e83125.

Anjum, S.I., Shah, A.H., Aurungzeb, M., Kori, J., Azim, M.K., Ansari, M.J. & Bin, L. (2018). Characterization of gut bacterial flora of Apis mellifera from north-west Pakistan. Saudi Journal of Biological Sciences, 25: 388-392.

Cai, J., Yang, H., Shi, S., Zhong, G. & Yi, X. (2018). Behavioural, morphological, and gene expression changes induced by 60Co-γ ray irradiation in Bactrocera tau (Walker). Frontiers in Physiology, 9: 118.

Chambers, J.M. (2008). Software for data analysis: programming with R. Springer, New York.

Coleman, J., Juhn, J. & James, J.J. (2006). Dissection of midgut and salivary glands from Ae. aegypti mosquitoes. Journal of Visualized Experiments, 5: 228.

Delfinado-Baker, M., Baker, E.W. & Phoon, A.C.G. (1989). Mites (Acari) associated with bees (Apidae) in Asia, with description of a new species. American Bee Journal, 129: 609–613.

Dillon, R.J., Webster, G., Weightman, A.J. & Keith Charnley, A. (2010). Diversity of gut microbiota increases with aging and starvation in the desert locust. Antonie Van Leeuwenhoek, 97: 69-77.

Disayathanoowat, T., Yoshiyama, M., Kimura, K. & Chantawannakul, P. (2015). Isolation and characterization of bacteria from the midgut of the Asian honey bee (Apis cerana indica). Journal of Apicultural Research, 51: 312-319.

Divya, K.K., Amritha, V.S., Viswanathan, A. & Devanesan, S. (2020). Microbial Count in Stingless Bee Honey (Tetragonula Iridipennis (Smith)). Life Science Advances, 2394.

Ellegaard, K.M. & Engel, P. (2019). Genomic diversity landscape of the honey bee gut microbiota. Nature Communications, 10: 1-13.

Feldhaar, H. (2011). Bacterial symbionts as mediators of ecologically important traits of insect hosts. Ecological Entomology, 36: 533-543.

Forsgren, E., Olofsson, T.C., Vasquez, A. & Fries, I. (2010). Novel lactic acid bacteria inhibiting Paenibacillus larvae in honey bee larvae. Apidologie, 41: 99-108.

Grover, M., Nain, L. & Saxena, A.K. (2009). Comparision between Bacillus subtilisRP24 and its antibiotic-defective mutants. World Journal of Microbiology and Biotechnology, 25: 1329-1335.

Gupta, A. & Nair, S. (2020). Dynamics of insect–microbiome interaction influence host and microbial symbiont. Frontiers in Microbiology, 11: 545024.

Kroiss, J., Kaltenpoth, M., Schneider, B., Schwinger, M.G., Hertweck, C., Maddula, R.K., Strohm E. & Svatos, A. (2010). Symbiotic streptomycetes provide antibiotic combination prophylaxis for wasp offspring. Nature Chemical Biology, 6: 261-263.

Kumar, S., Stecher, G. & Tamura, K. (2016). MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33: 1870-1874.

Kwong, W.K & Moran, N.A. (2016). Gut microbial communities of social bees. Nature Reviews Microbiology, 14: 374–384.

Kwong, W.K., Medina, L.A., Koch, H., Sing, K.S., Soh, E.J.Y., Ascher, J.S., Jaffe, R. & Moran, N.A. (2017). Dynamic microbiome evolution in social bees. Science Advances, 3: e1600513.

Leonhardt, S.D. & Kaltenpoth, M. (2014). Microbial communities of three sympatric Australian stingless bee species. PLoS ONE, 9: e105718.

Li, J., Qin, H., Wu, J., Sadd, B.M., Wang, X., Evans, J.D., et al., (2012). The prevalence of parasites and pathogens in Asian honeybees Apis cerana in China. PLoS ONE, 7: e47955.

Mohammad, S.M., Mahmud-Ab-Rashid, N.K. & Zawawi, N. (2020). Probiotic properties of bacteria isolated from bee bread of stingless bee Heterotrigona itama. Journal of Apicultural Research, 60: 172-87.

Ngalimat, M.S., Rahman, R.N.Z.R.A., Yusof, M.T., Syahir, A. & Sabri, S. (2019). Characterisation of bacteria isolated from the stingless bee, Heterotrigona itama, honey, bee bread and propolis. Peer J, 7: e7478.

Pajor, M., Worobo, R.W., Milewski, S. & Szweda, P. (2018). The antimicrobial potential of bacteria isolated from honey samples produces in the apiaries located in Pomeranian Voivodeship in Northern Poland. International Journal of Environmental Research and Public Health, 15: 2002.

Prabhakar, C.S., Sood, P., Kanwar, S.S., Sharma, P.N., Kumar, A. & Mehta, P.K. (2013). Isolation and characterization of gut bacteria of fruit fly, Bactrocera tau (Walker). Phytoparasitica, 41: 193-201.

Pucciarelli, A.B., Schapovaloff, M.E., Kummritz, S., Señuk, I.A., Brumovsky, L.A. & Dallagnol, A.M. (2014). Microbiological and physicochemical analysis of Yateí (Tetragonisca angustula) honey for assessing quality standards and commercialization. Revista Argentina de Microbiologia, 46: 325-332.

Romiguier, J., Cameron, S.A., Woodard, S.H., Fischman, B.J., Keller, L. & Praz, C.J. (2016). Phylogenomics controlling for base compositional bias reveals a single origin of eusociality in corbiculate bees. Molecular Biology and Evolution, 33: 670-678.

Singh, S., Singh, A., Baweja, V., Roy, A., Chakraborty, A. & Singh, I.K. (2021). Molecular rationale of insect-microbes symbiosis-from insect behaviour to mechanism. Microorganisms, 9: 2422.

Tamura, K., Nei, M. & Kumar, S. (2004). Prospects for inferring very large phylogenies by using the neighbour-joining method. Proceedings of the National Academy of Sciences U.S.A., 101: 11030-11035.

Tamura, K., Stecher, G. & Kumar, S. (2021). MEGA 11: Molecular evolutionary genetics analysis Version 11. Molecular Biology and Evolution, 37: 3022-3027.

Vásquez, A., Forsgren, E., Fries, I., Paxton, R.J., Flaberg, E., Szekely, L. & Olofsson, T.C. (2012), Symbionts as major modulators of insect health: lactic acid bacteria and honeybees. PLoS ONE, 7: e33188.

Wisselink, M., Aanen, D.K. & Van’t Padje, A. (2020). The longevity of colonies 455 of fungus-growing termites and the stability of the symbiosis. Insects, 11: 527.

Wright, M.H., Adelskov, J. & Greene, A.C. (2017). Bacterial DNA extraction using individual enzymes and phenol/chloroform separation. Journal of Microbiology and Biology Education, 18: 10-1128.

Yaacob, S.N.S., Raja, F., Ibrahim, R.K. & Wahab, R.A. (2018). Identification of Lactobacillus spp. and Fructobacillus spp. isolated from fresh Heterotrigona itama honey and their antagonistic activities against clinical pathogenic bacteria. Journal of Apicultural Research, 57: 395-405.

Yilmaz, M., Soran, H. & Beyatli, Y. (2006). Antimicrobial activities of some Bacillus spp. strains isolated from soil. Microbiological Research, 161: 127-131.

Zulkhairi Amin, F.A., Sabri, S., Ismail, M., Chan, K.W., Ismail, N., Mohd Esa, N. et al. (2020). Probiotic properties of Bacillus strains isolated from stingless bee (Heterotrigona itama) honey collected across Malaysia. International Journal of Environmental Research and Public Health, 17: 278.

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Published

2025-07-11

How to Cite

Vignesh, B. S., Jayaraj, J., Nalini, R., Kumutha, K., Srinivasan, M. R., Jayakanthan, M., Suresh, K., & Sabatina, P. (2025). Unveiling the Cultivation-Dependent Gut Bacterial Microbiota in the Feral Stingless Bee, Tetragonula iridipennis Smith (hymenoptera: Apidae). Sociobiology, 72(3), e11205. https://doi.org/10.13102/sociobiology.v72i3.11205

Issue

Vol. 72 No. 3 (2025)

Section

Research Article - Bees

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Copyright (c) 2025 B. Saai Vignesh, J. Jayaraj, R. Nalini, K. Kumutha, M. R. Srinivasan, M. Jayakanthan, K. Suresh, P. Sabatina

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