Effect of rotenone on the gut microbiota of Blattella germanica

doi:10.11853/j.issn.1003.8280.2022.05.012

Abstract

Abstract: Objective To investigate the effect of rotenone on the gut microbiota of Blattella germanica and the reason for the low toxicity of rotenone against B.germanica.Methods The gut microbiota of B.germanica were eliminated by feeding antibiotics,and then normal B.germanica cockroaches and the B.germanica cockroaches with eliminated gut microbiota were fed with rotenone to observe the influence of gut microbiota on the insecticidal activity of rotenone.In addition,gut genomic DNA was extracted from normal B.germanica cockroaches after feeding rotenone,and normal B.germanica cockroaches not fed with rotenone were used as control.Illumina HiSeq high-throughput sequencing was used to determine the sequence of the V3-V4 variable region of 16S rRNA,and the number of sequences and operational taxonomic units were used to analyze the composition and abundance of microflora,the abundance and diversity of bacterial communities,and the changes in bacterial composition across different treatment methods.SPSS 22.0 software was used for data analysis,and the independent samples t-test was used for difference analysis.R language software was used for heatmap clustering analysis and principal component analysis.Results Compared with the normal B.germanica cockroaches,the B.germanica cockroaches with eliminated gut microbiota had a significant reduction in survival rate after treatment with rotenone (t=8.485,P<0.001).16S rRNA sequencing obtained 128 genera,78 families,and 19 phyla of bacteria.Diversity analysis showed that the B.germanica cockroaches treated with rotenone had significantly higher ACE and Chao1 indices than the normal cockroaches (t=-2.990,P=0.040;t=-4.227,P=0.013).The analysis of bacterial community abundance showed that compared with the normal B.germanica cockroaches,the B.germanica cockroaches treated with rotenone had significantly higher relative abundances of Mitochondria,Bacteroidaceae,and Rikenellaceae (t=-12.178,P<0.001;t=-4.087,P=0.014;t=-3.570,P=0.026) and significantly lower relative abundances of Lachnospiraceae,Ruminococcaceae,Syntrophomonadaceae,Caulobacteraceae,and Enterobacteriaceae (t=10.662,P<0.001;t=8.242,P=0.001;t=4.394,P=0.012;t=3.421,P=0.025;t=3.038,P=0.041).Heatmap clustering analysis showed that the rotenone treatment group and the control group were in different primary branches,and the principal component analysis showed similar bacterial diversity between the samples in the same treatment.Conclusion Gut microbiota may play an role in host resistance to rotenone toxicity,and rotenone has a great impact on the community structure of the gut microbiota of B.germanica.Improving insecticidal activity by destroying the community structure of the gut microbiota of insects may provide a new idea for the application of plant-derived pesticides in pest control.

Key words: Blattella germanica, Rotenone, Gut microbiota, 16S rRNA

摘要： 目的明确鱼藤酮对德国小蠊肠道微生物的影响，分析鱼藤酮对德国小蠊胃毒活性低的原因。方法通过饲喂抗生素清除德国小蠊肠道微生物，然后用鱼藤酮分别饲喂正常德国小蠊和肠道微生物被清除的德国小蠊，检测肠道微生物对鱼藤酮杀虫活性的影响。正常德国小蠊饲喂鱼藤酮后提取肠道基因组DNA，未饲喂鱼藤酮的正常德国小蠊作为对照，利用Illumina Hiseq高通量测序技术测定16S rRNA V3－V4可变区域序列，整理统计测得的序列数目和操作分类单元，分析细菌菌群的组成和丰度、细菌群落的丰富度和多样性，以及菌群组成在不同处理组间的变化。采用SPSS 20.0软件进行数据分析，选取独立样本t检验，利用R语言做聚类热图及主成分分析。结果与正常德国小蠊相比，肠道微生物被清除的德国小蠊饲喂鱼藤酮后，其存活率显著降低（t=8.485，P<0.001）。16S rRNA测序共检测到19门78科128属的细菌。多样性分析结果表明，鱼藤酮处理后德国小蠊肠道微生物ACE指数和Chao1指数均高于对照组（t=-2.990，P=0.040；t=-4.227，P=0.013）。细菌群落丰度结果显示，鱼藤酮处理后德国小蠊线粒体菌科、拟杆菌科和理研菌科的相对丰度均高于对照组（t=-12.178，P<0.001；t=-4.087，P=0.014；t=-3.570，P=0.026）；毛螺菌科、瘤胃菌科、单胞菌科、柄细菌科及肠杆菌科的相对丰度则均低于对照组（t=10.662，P<0.001；t=8.242，P=0.001；t=4.394，P=0.012；t=3.421，P=0.025；t=3.038，P=0.041）。聚类热图分析的结果显示鱼藤酮处理组和对照组处于不同的初级分支，主成分分析可见同一处理的样本间细菌多样性相近。结论肠道微生物可能在宿主抵御鱼藤酮毒性过程中起一定作用，鱼藤酮对德国小蠊肠道微生物群落结构有较大影响。破坏昆虫肠道微生物群落结构提高杀虫剂杀虫活性可能为植物源农药在害虫防治上的应用提供新的思路。

关键词: 德国小蠊, 鱼藤酮, 肠道微生物, 16S rRNA

CLC Number:

ZHU Jian, CHEN Guang-ming, WU Ying-hua, LIAO Guo-dong, XU Han-hong. Effect of rotenone on the gut microbiota of Blattella germanica[J]. Chinese Journal of Vector Biology and Control, 2022, 33(5): 677-683.

朱剑, 陈光明, 吴鹰花, 廖国栋, 徐汉虹. 鱼藤酮对德国小蠊肠道微生物的影响[J]. 中国媒介生物学及控制杂志, 2022, 33(5): 677-683.

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