江西地区野猪菌群多样性分析及其携带产气荚膜梭菌的分离鉴定

doi:10.11843/j.issn.0366-6964.2025.04.035

摘要/Abstract

摘要：

旨在探究野猪呼吸道、肠道及粪便中的菌群组成和携带产气荚膜梭菌(Clostridium perfringens，Cp)的相关情况。采集江西地区27头野猪呼吸道、肠道及粪便样品，采用细菌16S rRNA基因测序技术分析野猪不同生理部位的菌群组成和差异，分析野猪源Cp基因型及毒素基因cpa、cpb、cpb2、etx、itx、cpe和netb的携带情况，并分离纯化肠道与粪便样品中的产气荚膜梭菌，对分离株进行全基因组测序分析。菌群分析结果显示：三组的优势菌门都为厚壁菌门、变形菌门、拟杆菌门和放线菌门，其中肠道和粪便中厚壁菌门的占比最高，呼吸道中变形菌门的占比最高。差异性分析发现三组中5个菌属存在极显著差异，其余10个菌属存在显著差异；从野猪粪便样品中分离到了三株Cp，经PCR毒素基因分型检测和全基因组测序分析确认为A型，并且无β2毒素基因；27份样品中Cp的携带率为81.48%(22/27)，在这22份阳性样品中，β2毒素基因的检出率为9份，占40.91%(9/22)。其它毒素基因未检测到，综合各分型毒素检测结果，阳性样品中存在的Cp均为A型。综上，本研究基于菌群分析，了解了野猪不同生理部位的菌群组成，以及Cp在野猪中的携带情况，为预防、检测野猪源的相关疾病提供了理论参考。

关键词: 野猪, 菌群分析, 产气荚膜梭菌

Abstract:

The aim of this study was to investigate the composition of the flora in the respiratory, intestinal and fecal tract of wild boars and the carriage situation of Clostridium perfringens (Cp). Respiratory, intestinal and fecal samples from twenty-seven wild boars were collected in Jiangxi region. Bacterial 16S rRNA gene sequencing technology was used to analyze the composition and differences of the flora in different physiological parts of wild boars. And the Cp genotypes of wild boar sources and the carriage of toxin genes cpa, cpb, cpb2, etx, itx, cpe and netb were also analyzed. The Clostridium perfringens in the intestinal and fecal samples were isolated and purified, in additon, the whole-genome sequencing were conducted on the isolates. The results of the flora analysis showed that the dominant phyla in all three groups were Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria, with the highest percentage of Firmicutes in the intestinal tract and feces and the highest percentage of Proteobacteria in the respiratory tract. Analysis of variance revealed highly significant differences in five genera in the three groups and significant differences in the remaining 10 genera; three Cp strains isolated from wild boar fecal samples were confirmed to be type A and were free of β2 toxin gene by PCR toxin genotyping and whole genome sequencing analysis; the carriage rate of Cp was 81.48% (22/27) in 27 samples, and the strains carried β2 toxin gene in these 22 positive samples were 9, representing 40.91% (9/22). No other toxin genes were detected. Combining the results of the toxin tests for each subtype, the Cp presented in the positive samples were all type A. In conclusion, based on the flora analysis, this study understood the microbial community composition of different physiological parts of wild boars and the carriage of Cp in wild boar, which provided theoretical clues for the prevention and detection of wild boar diseases.

Key words: wild boar, microbial community analysis, Clostridium perfringens

中图分类号:

S858.9

谭娟娟, 杨贝莹, 武前悦, 花慧颖, 曹华斌, 严珲, 张锦华. 江西地区野猪菌群多样性分析及其携带产气荚膜梭菌的分离鉴定[J]. 畜牧兽医学报, 2025, 56(4): 1876-1886.

TAN Juanjuan, YANG Beiying, WU Qianyue, HUA Huiying, CAO Huabin, YAN Hui, ZHANG Jinhua. Diversity Analysis of Wild Boar Flora in Jiangxi Region and the Isolation and Identification of Clostridium perfringens Carried by Wild Boar[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1876-1886.

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采样地区 Sampling area	采样数 Number of samples	样品类型 Sample type
采样地区 Sampling area	采样数 Number of samples	肠道 Intestine	粪便 Feces	呼吸道 Respiratory tract
南昌市安义县 Anyi County, Nanchang City	2	2	2	0
南昌市新建区 Xinjian District, Nanchang City	3	2	3	0
南昌市瑶湖区 Yaohu District, Nanchang City	1	1	0	0
九江市武宁县 Wuning County, Jiujiang City	6	6	6	0
九江市修水县 Xiushui County, Jiujiang City	15	15	15	15
总计 Total	27	27	26	15

基因 Gene	序列(5′→3′) Sequence	片段大小/bp Sequence size
cpa	GCTAATGTTACTGCCGTTGA CCTCTGATACATCGTGTAAG	324
cpb	GCGAATATGCTGAATCATCTA GCAGGAACATTAGTATATCTTC	196
etx	GCGGTGATATCCATCTATTC CCACTTACTTGTCCTACTAAC	655
cpe	GGAGATGGTTGGATATTAGG GGACCAGCAGTTGTAGATA	233
itx	ACTACTCTCAGACAAGACAG CTTTCCTTCTATTACTATACG	446
netb	GCTGGTGCTGGAATAAATGC TCGCCATTGAGTAGTTTCCC	384
atyβ2	ATTATGTTTAGGAATACAGTTA CAATACCCTTCACCAAATACTC	750
conβ2	CAATTGGGGGAGTTTATCCACAA CAATACCCTTCACCAAATACTC	300

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