昆虫学报 ›› 2024, Vol. 67 ›› Issue (9): 1190-1199.doi: 10.16380/j.kcxb.2024.09.003

• 研究论文 • 上一篇    下一篇

美洲大蠊粪便中微生物的分离和鉴定及其抗菌活性分析

官婷婷, 许文慧, 肖光帅, 杨大松*, 杨银河*   

  1. (大理大学药学院, 云南省昆虫生物医药研发重点实验室, 大理 671000)
  • 出版日期:2024-09-20 发布日期:2024-10-22

Isolation and identification of microorganisms from Periplaneta americana (Dictyoptera: Blattidae) frass and analysis of their antibacterial activity

GUAN Ting-Ting, XU Wen-Hui, XIAO Guang-Shuai, YANG Da-Song*, YANG Yin-He*   

  1.  (Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D, College of Pharmacy, Dali University, Dali 67100, China)
  • Online:2024-09-20 Published:2024-10-22

摘要: 【目的】探究美洲大蠊Periplaneta americana粪便中具有抗菌活性的真菌和放线菌,为抗菌药物的开发提供微生物来源。【方法】利用稀释涂布法与选择培养法,从云南省大理白族自治州巍山县美洲大蠊养殖场收集的美洲大蠊成虫粪便中分离纯化放线菌和真菌;采用牛津杯琼脂扩散法检测菌株代谢产物对6种病原细菌大肠杆菌Escherichia coli、粪肠球菌Enterococcus faecalis、金黄色葡萄球菌Staphylococcus aureus、 耐甲氧西林的金黄色葡萄球菌(methicillin-resistant S. aureus)、 铜绿假单胞菌Pseudomonas aeruginosa和鼠伤寒沙门氏菌Salmonella typhimurium的抗菌活性; 通过16S rDNA和ITS rDNA基因测序鉴定抗菌活性菌株,并利用邻接法构建抗菌活性菌株系统发育树。【结果】从美洲大蠊成虫粪便中分离出真菌17株,放线菌19株。共有6株真菌和12株放线菌表现出不同程度的抗菌活性,其中真菌EPAF15-EPAF17及放线菌EPAA12和EPAA17对大肠杆菌、金黄色葡萄球菌、耐甲氧西林的金黄色葡萄球菌、铜绿假单胞菌和鼠伤寒沙门氏菌表现出较为广谱的抗菌活性。系统发育分析显示,具有抗菌活性的真菌隶属于镰刀菌属Fusarium、生丝毕赤酵母属Hyphopichia和丝孢酵母属Trichosporon;具有抗菌活性的放线菌隶属于链霉属Streptomyces。【结论】美洲大蠊成虫粪便中具有种类丰富的可培养微生物,其中的抗菌活性真菌和放线菌的发现不仅能作为进一步深入挖掘新型抗菌活性天然药物的重要微生物储备资源,也为特境微生物资源的充分利用提供了新的思路。

关键词: 美洲大蠊, 粪便, 真菌, 放线菌, 抗菌活性, 分子鉴定, 系统发育

Abstract: 【Aim】 To investigate the fungi and actinomycetes with antimicrobial activity in Periplaneta americana frass, so as to supply microbial sources for the development of antimicrobial medications. 【Methods】 Using the dilution coating method and selective culture method, actinomycetes and fungi were isolated and purified from the frass of adult P. americana collected from the P. americana farm in Weishan County, Dali Bai Autonomous Prefecture, Yunnan Province, China. The antibacterial activities of metabolites of these strains against six pathogenic bacteria Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant S. aureus, Pseudomonas aeruginosa and Salmonella typhimurium were detected by the Oxford cup agar diffusion method. The strains with antibacterial activity were identified by the 16S rDNA gene sequencing and ITS rDNA gene sequencing, and a phylogenetic tree of the strains with antibacterial activities was constructed using the neighbor-joining method. 【Results】 Seventeen strains of fungi and 19 strains of actinomycetes were isolated from the frass of adult P. americana. Six strains of fungi and 12 strains of actinomycetes exhibited varying degrees of antibacterial activity. Among them, the fungi EPAF15-EPAF17 and the actinomycetes EPAA12 and EPAA17 showed broad-spectrum antibacterial activities against E. coli, S. aureus, methicillin-resistant S. aureus, P. aeruginosa and S. typhimurium. Phylogenetic analysis showed that the fungi with antibacterial activity belong to Fusarium, Hyphopichia and Trichosporon, and the actinomycetes with antibacterial activity belong to the genus Streptomyces. 【Conclusion】 There are various types of culturable microorganisms in the frass of adult P. americana. The discovery of fungi and actinomycetes with antibacterial activity provides essential microbial resources for further exploration of new antimicrobial natural drugs. It also introduces a new concept for fully utilizing microbial resources in a special environment.

Key words: Periplaneta americana, frass, fungi, actinomyces, antibacterial activity, molecular identification, phylogeny