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

B. Saai Vignesh; J. Jayaraj; R. Nalini; K. Kumutha; M. R. Srinivasan; M. Jayakanthan; K. Suresh; P. Sabatina

doi:10.13102/sociobiology.v72i3.11205

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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