孟加拉虎体表不同饱血状态长角血蜱中肠菌群结构的分析

doi:10.11843/j.issn.0366-6964.2025.12.040

摘要/Abstract

摘要：

旨在探究孟加拉虎体表不同饱血状态长角血蜱雌蜱中肠菌群结构的特点。本研究采集孟加拉虎体表饱血(fully engorged，FF)与半饱血(half engorged，HF)长角血蜱雌蜱，在无菌条件下收集各组蜱中肠内容物，提取各组样本细菌总DNA，PCR扩增16S rRNA V3-V4区，构建文库，Illumina NovaSeq平台双末端测序，测序数据经拼接、过滤、降噪后，进行Amplicon Sequence Variant(ASV)和多样性分析。结果显示，FF、HF获得的有效序列分别为100 021条和82 657条，聚类后FF、HF分别获得ASVs为259、541个，两组样品共有的ASVs为116个。在门水平上，两组样本均以变形菌门、厚壁菌门、拟杆菌门、放线菌门为优势菌门；变形菌门在FF中的含量大于HF，其它三个优势菌门反之。在属水平上，柯克斯氏体属、Muribaculaceae、棒状杆菌属、代尔夫特菌属、葡萄球菌属为FF和HF中的共有优势菌属，其中柯克斯氏体属在两组样本中含量均较高，且在FF中的含量高于HF；FF、HF中特有且相对丰度较高的菌属分别为Schlegelella、普雷沃氏菌属等。在种水平上，两组样本以柯克斯氏体属内共生体含量最高，其在FF和HF中的分布特点与柯克斯氏体属在两组样本中的分布特点相一致。综上，长角血蜱雌蜱中肠微生物菌群结构复杂，不同饱血状态雌蜱中肠菌群结构存在一定差异，且同一菌属相对丰度不同。

关键词: 长角血蜱, 不同饱血状态, 中肠菌群结构, 蜱及蜱传病, 高通量测序

Abstract:

This study aimed to investigate the microflora structural characteristics of the midgut of different engorged Haemaphysalis longicornis(H. longicornis) females collected from Panthera tigris tigris. Female H. longicornis in fully engorged (FF) and half engorged (HF) were sampled, and midgut contents were collected from each group of ticks under aseptic conditions. Total bacterial DNA was extracted from each group of samples. The V3-V4 areas of 16S rRNA was amplified by PCR, and the libraries were constructed for each group. Paired-end sequencing was performed using the Illumina NovaSeq platform. Sequencing data were processed by merging, filtering, and denoising, followed by Amplicon Sequence Variant (ASV) clustering and diversity analysis. The results showed that 100 021 and 82 657 effective tags were obtained from FF and HF, respectively. The number of ASVs obtained from FF and HF after clustering was 259 and 541, respectively, of which 116 were common between the two groups. At the phylum level, the dominant common phyla were Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteriota in both groups, with Proteobacteria being more abundant in FF, while the other three dominant phyla were relatively more abundant in HF. At the genus level, the dominant common genera were Coxiella, Muribaculaceae, Corynebacterium, Delftia, and Staphylococcus in both groups. Coxiella had the highest relative abundance, and the relative abundance of Coxiella was higher in FF than in HF. In FF, genera such as Schlegelella were unique and relatively abundant, while in HF, genera such as Prevotella were unique and relatively abundant. At the species level, Coxiella endosymbionts were the most abundant taxa in both groups, with distribution patterns consistent with Coxiella. The results suggested that the midgut of female H. longicornis has a complex microflora. There were certain differences in the midgut microflora of different engorged ticks, and the relative abundance of the same genus varied in different samples.

Key words: Haemaphysalis longicornis, different engorged statuses, midgut microbial community structure, ticks and tick-borne diseases, high-throughput sequencing

中图分类号:

S852.746

刘潇雨, 李勇, 邱启官, 程天印, 段德勇. 孟加拉虎体表不同饱血状态长角血蜱中肠菌群结构的分析[J]. 畜牧兽医学报, 2025, 56(12): 6386-6396.

LIU Xiaoyu, LI Yong, QIU Qiguan, CHENG Tianyin, DUAN Deyong. Analysis of the Midgut Microflora of Different Engorged Haemaphysalis longicornis Collected from Panthera tigris tigris[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6386-6396.

图/表 6

图 1

表 1

图 2

图 3

表 2

两组样本中前15个共有细菌属以外其他共有细菌属的相对丰度"

细菌属 Bacterial genus	细菌属在每个样本中的相对丰度 Relative abundance of bacteria genus in each sample
细菌属 Bacterial genus	FF	HF
副拟杆菌属Parabacteroides	0.000 026 508	0.004 718 482
罗姆布茨菌属Romboutsia	0.001 192 875	0.004 387 128
Lachnoclostridium	0.000 039 762	0.00 331 354
梭状芽孢杆菌属Clostridium sensu stricto	0.000 119 287	0.003 207 507
真杆菌属Eubacterium	0.000 026 508	0.002 809 882
毛螺菌科NK4A136属Lachnospiraceae NK4A136	0.000 675 962	0.002 783 374
Rikenellaceae RC9 gut group	0.000 198 812	0.002 107 412
罗斯氏菌属Rothia	0.002 054 395	0.000 384 371
短杆菌属Brevibacterium	0.001 047 079	0.001 802 566
短状杆菌属Brachybacterium	0.000 861 521	0.001 723 041
肠球菌属Enterococcus	0.001 577 245	0.000 092 779
Anaerotruncus	0.000 543 421	0.00 149 772
Incertae sedis	0.000 119 287	0.001 259 145
螺杆菌属Helicobacter	0.001 179 620	0.000 106 033
微球菌属Micrococcus	0.000 344 608	0.001 153 112
苏黎世杆菌属Turicibacter	0.00 015 905	0.000 980 808
Dysgonomonas	0.000 013 254	0.000 848 266
另枝菌属Alistipes	0.000 556 675	0.000 622 946
阿克曼氏菌属Akkermansia	0.000 013 254	0.000 583 183
无色杆菌属Achromobacter	0.000 530 166	0.000 424 133
Cutibacterium	0.000 490 404	0.000 172 304
Eubacterium xylanophilum group	0.000 106 033	0.000 463 896
不动杆菌属Acinetobacter	0.000 384 371	0.000 463 896
A2	0.000 039 762	0.000 450 642
GCA-900066575	0.000 039 762	0.000 437 387
毛螺菌科UCG-006属Lachnospiraceae UCG-006	0.000 079 525	0.000 397 625
溶杆菌属Lysobacter	0.000 185 558	0.000 384 371
Clostridia vadinBB60 group	0.000 371 117	0.000 357 862
巨大球菌属Macrococcus	0.000 053 017	0.0 003 181
Lawsonella	0.000 291 592	0.000 265 083
布兰汉氏菌属Alkanindiges	0.000 039 762	0.000 291 592
Colidextribacter	0.000 092 779	0.000 238 575
栖水菌属Enhydrobacter	0.000 092 779	0.000 225 321
Deep Sea Euryarchaeotic Group	0.000 119 287	0.000 092 779
食酸菌属Acidovorax	0.000 026 508	0.000 106 033
ASF356	0.000 053 017	0.000 106 033
考克氏菌属Kocuria	0.000 039 762	0.000 092 779

表 2

表 3

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样本 Sample	测序覆盖率 Good’s coverage	Chao1多样性指数 Chao1	香农多样性指数 Shannon	辛普森多样性指数 Simpson
HF	1.000	541.000	3.737	0.635
FF	1.000	259.375	1.519	0.290

细菌种 Bacterial species	相对丰度 Relative abundance	细菌种 Bacterial species	相对丰度 Relative abundance
Clostridium aldenense	0.001 219 383	bacterium XPD3003	0.000 146
Blattella germanica	0.001 153 112	Marinococcus sp.	0.000 133
产琥珀酸丝状杆菌Fibrobacter succinogenes	0.000 781 996	Luteimonas sp.	0.000 119
Dorea sp.	0.000 649 454	Chryseobacterium salipaludis	0.000 119
栖瘤胃普雷沃氏菌Prevotella ruminicola	0.000 583 183	bacterium enrichment	0.000 106
Bacteroides sp.	0.000 490 404	Treponema bryantii	0.000 106
bacterium AC2043	0.000 490 404	bacterium YRD2003	0.000 106
rumen bacterium	0.000 424 133	Azospirillum sp.	0.000 093
Fibrobacter sp.	0.000 397 625	bacterium ND2018	0.000 093
梭菌目细菌Clostridiales bacterium	0.000 344 608	bacterium MD2012	0.000 066
Ruminococcus sp.	0.000 291 592	bacterium YGB2004	0.000 066
黄化瘤胃球菌Ruminococcus flavefaciens	0.000 278 337	Chryseobacterium chengduensis	0.000 066
拟杆菌目细菌Bacteroidales bacterium	0.000 278 337	rumen bacterium	0.000 066
Adlercreutzia mucosicola	0.000 212 067	bacterium MB2022	0.000 066
rumen bacterium	0.000 185 558	bacterium V9D2012	0.000 066
汤氏不动杆菌Acinetobacter towneri	0.000 172 304	布雷恩特普雷沃菌Prevotella bryantii	0.000 066
bacterium V9D2002	0.00 015 905
bacterium WCE2007	0.000 159

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