中国血吸虫病防治杂志 ›› 2022, Vol. 34 ›› Issue (5): 475-.

• 论著 • 上一篇    下一篇

白纹伊蚊不同发育阶段体内菌群结构及多样性分析

赵永俏1, 2,夏骜1, 2,张梅花2,李菊林2,朱国鼎1, 2,唐建霞1, 2*   

  1. 1 南京医科大学公共卫生学院(江苏 南京 211166);2 国家卫生健康委员会寄生虫病预防与控制技术重点实验室、江苏省寄生虫与媒介控制技术重点实验室、江苏省血吸虫病防治研究所(江苏 无锡 214064)
  • 出版日期:2022-11-23 发布日期:2022-11-23
  • 作者简介:赵永俏,女,硕士研究生。研究方向:媒介生物学
  • 基金资助:
    国家重点研发基金(2020YFC1200105);国家自然科学基金(82172302)

Microbiota structure and diversity in Aedes albopictus at different developmental stages

ZHAO Yong⁃qiao1, 2, XIA Ao1, 2, ZHANG Mei⁃hua2, LI Ju⁃lin2, ZHU Guo⁃ding1, 2, TANG Jian⁃xia1, 2*   

  1. 1 School of Public Health, Nanjing Medical University, Nanjing, Jiangsu 211166, China; 2 Key Laboratory of National Health Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu 214064, China
  • Online:2022-11-23 Published:2022-11-23

摘要: 目的 了解实验室品系白纹伊蚊不同发育阶段体内及幼虫生存水体中菌群多样性及其构成。方法 收集实验室品系白纹伊蚊幼虫生存水体及不同发育阶段伊蚊,通过Illumina Miseq二代高通量测序技术对菌群16S 核糖体RNA(16S ribosomal RNA,16S rRNA)基因V3和V4区进行测序。采用alpha多样性指数分析白纹伊蚊不同发育阶段菌群丰度和多样性,采用主成分分析对不同发育阶段菌群结构进行差异分析,采用菌群构成柱状图和Venn图表示白纹伊蚊不同发育阶段菌群组成结构和相对丰度。结果 白纹伊蚊幼虫生存水体及各个发育阶段共检测到16个门、30个纲、72个目、129个科和224个属细菌。多样性指数显示,白纹伊蚊幼虫期菌群多样性最高(Chao指数为125.20 ± 30.48、Shannon指数为2.04 ± 0.39),与其生存水体中的菌群多样性(Chao指数为 118.52 ± 15.07、Shannon指数为2.15 ± 0.30)相似(t = 0.35、-0.41,P均> 0.05);雌性成蚊体内菌群丰度和均匀度(Chao指数为 42.50 ± 3.54、Shannon指数为1.25 ± 1.67)均高于雄蚊(Chao指数为18.50 ± 2.13,Shannon指数为0.50 ± 0.05)(t = 8.23、6.00,P均< 0.05)。在门水平上,变形菌门、拟杆菌门、放线菌门和厚壁菌门细菌为白纹伊蚊各个发育阶段所共有,其中蛹期以变形菌门居多,占90.36%;成蚊期以拟杆菌门居多,其在雌蚊和雄蚊体内菌群中分别占46.01%和86.11%;放线菌门是幼虫期菌群的重要组成部分,占32.10%。在属水平上,白纹伊蚊各发育阶段共同优势菌属为伊丽莎白菌属和拉恩氏菌属;其中拉恩氏菌属细菌是蛹期的主要组成部分,占87.56%;伊丽莎白菌属细菌是成蚊期的主要组成部分,在雌蚊和雄蚊体内分别占46.01%和86.11%;幼虫中微杆菌属细菌占比最高,为12.11%。在种水平上,代尔夫特菌属、伊丽莎白菌属、罗姆布茨菌、沙雷氏菌属、拉恩菌属、肠球菌属、微杆菌属等7种菌属细菌为白纹伊蚊各发育阶段所共有。在白纹伊蚊幼虫期及其生存水体中的共有菌属中,土壤杆菌属和鞘氨醇杆菌属细菌占比最高,分别占17.54%和13.93%。结论 白纹伊蚊不同发育阶段菌群构成存在差异,但在门水平上保持了相似的菌群多样性,幼虫期体内菌群多样性与其生存水体中的菌群多样性相似。

关键词: 白纹伊蚊, 幼虫, 蛹, 成蚊, 菌群多样性, 16S 核糖体RNA

Abstract: Objective To investigate the diversity and composition of microflora in laboratory⁃reared Aedes albopictus at different developmental stages and larval habitat waters. Methods The larval habitat waters and different developmental stages of laboratory⁃reared A. albopictus were collected, and the V3 and V4 regions of the bacterial 16S rRNA gene were sequenced using Illumina Miseq next⁃generation sequencing. The abundance and diversity of the microflora were examined using alpha diversity index in A. albopictus at different developmental stages, and the difference in the microflora compositions was compared in A. albopictus at different developmental stages using principal component analysis (PCA). In addition, the species composition and relative abundance of microflora in A. albopictus at different developmental stages were described using histograms and Venn diagrams. Results A total of 16 phyla, 30 classes, 72 orders, 129 families and 224 genera of bacteria were detected in larval habitat waters and different developmental stages of A. albopictus. The highest bacterial diversity was seen in larval A. albopictus, with Chao index of 125.20 ± 30.48 and Shannon diversity index of 2.04 ± 0.39, which were comparable to those (Chao index of 118.52 ± 15.07 and Shannon diversity index of 2.15 ± 0.30) in larval habitat waters (t = 0.35 and -0.41, both P values > 0.05). The bacterial abundance and evenness were significantly greater in female adults than in male adults (Chao index: 42.50 ± 3.54 vs. 18.50 ± 2.13, t = 8.23, P < 0.05; Shannon diversity index: 1.25 ± 1.67 vs. 0.50 ± 0.05, t = 6.00, P < 0.05). Proteobacteria, Bacteroidota, Actinobacteriota and Firmicutes were four common phyla of bacteria at each developmental stage of A. albopictus, with Proteobacteria dominated at the pupal stage (90.36%), Bacteroidota dominated at the adult stage (46.01% in female adults and 86.11% in male adults), and Actinobacteriota dominated at the larval stage (32.10%). Elizabethkingia and Rahnella1 were common dominant genera of bacteria at each developmental stage of A. albopictus, with Rahnella1 as the major component at the pupal stage (87.56%), Elizabethkingia as the main component at the adult stage (46.01% in female adults and 86.11% in male adults, respectively), and Microbacteria as the dominant bacterial genus at the larval stage (12.11%). In addition, Delftia, Elizabethkingia, Romboutsia, Serratia, Rahnella1, Enterococcus and Microbacterium were common genera of bacteria at each developmental stage of A. albopictus, with Edaphobaculum dominated at the larval stage (17.54%) and Sphingobacterium dominated in larval habitat waters (13.93%). Conclusions There are differences in the composition of symbiotic bacteria at different developmental stages of A. albopictus; however, similar microflora diversity is maintained at the phylum level. The microflora diversity is comparable in larvae and larval habitat waters of A. albopictus.

Key words: Aedes albopictus, Larvae, Pupae, Adult mosquito, Microflora Diversity, 16S ribosomal RNA

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