Roy Nazish, Moon Sunmi, Kim Chaerin, Kim Jin-Myung, Lee Kwang-Sik, Shin Yongho, Shanmugam Gnanendra, Choi Kihyuck
Department of Applied Bioscience, Dong-A University, Busan, 49315, Republic of Korea.
Department of Biotechnology, Vivekanandha College of Arts and Sciences for Women (Autonomous), Tiruchengode, Tamilnadu, India.
Probiotics Antimicrob Proteins. 2025 Feb;17(1):51-61. doi: 10.1007/s12602-024-10248-w. Epub 2024 Apr 2.
To explore the potential of probiotic candidates beneficial for honeybee health through the modulation of the gut microbiome, bee gut microbes were isolated from bumblebee (Bombus terrestris) and honeybee (Apis mellifera) using diverse media and cultural conditions. A total of 77 bee gut bacteria, classified under the phyla Proteobacteria, Firmicutes, and Actinobacteria, were identified. The antagonistic activity of the isolates against Ascosphaera apis, a fungal pathogen responsible for chalkbrood disease in honeybee larvae, was investigated. The highest growth inhibition percentage against A. apis was demonstrated by Bacillus subtilis strain I3 among the bacterial strains. The presence of antimicrobial peptide genes in the I3 strain was detected using PCR amplification of gene fragments encoding surfactin and fengycin utilizing specific primers. The export of antimicrobial peptides by the I3 strain into growth medium was verified using liquid chromatography coupled with mass spectroscopy. Furthermore, the strain's capabilities for degrading pesticides, used for controlling varroa mites, and its spent growth medium antioxidant activity were substantiated. The survival rate of honeybees infected with (A) apis was investigated after feeding larvae with only medium (fructose + glucose + yeast extract + royal jelly), (B) subtilis I3 strain, A. apis with medium and I3 strain + A. apis with medium. Honeybees receiving the I3 strain + A. apis exhibited a 50% reduction in mortality rate due to I3 strain supplementation under experimental conditions, compared to the control group. In silico molecular docking revealed that fengycin hydrolase from I3 strain effectively interacted with tau-fluvalinate, suggesting its potential in bee health and environmental protection. Further studies are needed to confirm the effects of the I3 strain in different populations of honey bees across several regions to account for genetic and environmental variations.
为了通过调节肠道微生物群来探索对蜜蜂健康有益的益生菌候选物的潜力，利用多种培养基和培养条件从熊蜂（Bombus terrestris）和蜜蜂（Apis mellifera）中分离出蜜蜂肠道微生物。共鉴定出77种蜜蜂肠道细菌，分属于变形菌门、厚壁菌门和放线菌门。研究了这些分离菌株对蜜蜂幼虫白垩病致病真菌球囊菌（Ascosphaera apis）的拮抗活性。在这些细菌菌株中，枯草芽孢杆菌I3菌株对球囊菌的生长抑制率最高。利用特异性引物对编码表面活性素和丰原素的基因片段进行PCR扩增，检测I3菌株中抗菌肽基因的存在。采用液相色谱-质谱联用技术验证了I3菌株抗菌肽向生长培养基中的分泌。此外，还证实了该菌株降解用于防治蜂螨的农药的能力及其生长培养基的抗氧化活性。在用仅培养基（果糖+葡萄糖+酵母提取物+蜂王浆）、（B）枯草芽孢杆菌I3菌株、球囊菌与培养基以及I3菌株+球囊菌与培养基喂养幼虫后，研究了感染球囊菌的蜜蜂的存活率。在实验条件下，与对照组相比，接受I3菌株+球囊菌的蜜蜂由于补充了I3菌株，死亡率降低了50%。计算机模拟分子对接显示，I3菌株的丰原素水解酶与氟氯苯菊酯有效相互作用，表明其在蜜蜂健康和环境保护方面的潜力。需要进一步研究以确认I3菌株在不同地区不同蜜蜂种群中的作用，以考虑遗传和环境差异。
