Comparison of Surface Sterilization Methods for the Analysis of insect gut microbiota: Solenopsis invicta (Formicidae) as an example

IF 0.7 4区 农林科学 Q4 ENTOMOLOGY
Defu Chen, Zhonghao Huang, Youbang Li, Zhilin Chen
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Abstract

The gut microbiota of vertebrates and invertebrates has become the focus of recent research. However, current sterilizing practices need more systematic quantitative methods due to limitations caused by their minute body size. Thus, an effective sterilization process incorporating organic and inorganic methods to obtain invertebrate microbiota, particularly when evaluating smaller insects, has yet to be elucidated. This study investigated if the whole abdomen should be utilized as the material to sterilize and examined whether physical and chemical surface sterilization methods could be combined to facilitate the acquisition of gut microbiota from theimported red fire ant (Solenopsis invicta Buren, 1972). Eight methods were designed by incorporating three chemical reagents (sterile water, 2.0 mg/L NaClO, and 75% ethanol) and one physical treatment (250 nm UV). The length range of the amplified fragment in the red imported fire ant is 401-450 bp. According to the results of the GLM regression model and interaction effect model, none of these factors (sterile water, 2.0 mg/L NaClO, and 75% ethanol, 250 nm UV) were significant for statistical regression of the Chao index, and these factors did not significantly interact with each other. Based on Alpha and Beta diversity analysis, none of the methods significantly affected the diversity of insects’ gut microbiome. Finally, we suggested that it is feasible for different species of small insects to select appropriate methods according to the current situation. Still, it is best to achieve unity in the same group.
昆虫肠道菌群表面灭菌方法比较:以蚁科无尾螺梭菌(Solenopsis invicta)为例
脊椎动物和无脊椎动物的肠道微生物群已成为近年来研究的热点。然而,目前的灭菌实践需要更系统的定量方法,由于其微小的身体尺寸造成的限制。因此,结合有机和无机方法来获得无脊椎微生物群的有效灭菌过程,特别是在评估较小的昆虫时,尚未得到阐明。本研究探讨了是否应将整个腹部作为灭菌材料,并研究了物理和化学表面灭菌方法是否可以结合起来,以促进从进口红火蚁身上获取肠道微生物群(Solenopsis invicta Buren, 1972)。采用3种化学试剂(无菌水、2.0 mg/L NaClO和75%乙醇)和1种物理处理(250 nm UV)设计8种方法。该扩增片段的长度范围为401 ~ 450 bp。根据GLM回归模型和交互效应模型的结果,无菌水、2.0 mg/L NaClO和75%乙醇、250 nm UV对Chao指数的统计回归均不显著,且这些因素之间的交互作用不显著。基于Alpha和Beta多样性分析,没有一种方法显著影响昆虫肠道微生物组的多样性。最后,我们建议不同种类的小昆虫根据现状选择合适的方法是可行的。不过,最好还是在同一群体中实现团结。
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来源期刊
Sociobiology
Sociobiology 生物-昆虫学
CiteScore
1.60
自引率
11.10%
发文量
28
审稿时长
3 months
期刊介绍: SOCIOBIOLOGY publishes high quality articles that significantly contribute to the knowledge of Entomology, with emphasis on social insects. Articles previously submitted to other journals are not accepted. SOCIOBIOLOGY publishes original research papers and invited review articles on all aspects related to the biology, evolution and systematics of social and pre-social insects (Ants, Termites, Bees and Wasps). The journal is currently expanding its scope to incorporate the publication of articles dealing with other arthropods that exhibit sociality. Articles may cover a range of subjects such as ecology, ethology, morphology, population genetics, physiology, toxicology, reproduction, sociobiology, caste differentiation as well as economic impact and pest management.
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