猪链球菌SC124菌株中ICE_Prophage携带耐药基因的水平转移

doi:10.11843/j.issn.0366-6964.2024.08.041

摘要/Abstract

摘要：

旨在探究猪链球菌中ICE_Prophage携带耐药基因转移的情况，使用Illumina MiSeq和Pacbio RSII平台对猪链球菌SC124菌株进行了全基因组测序，利用在线网站RAST、ICEberg 3.0、PHASTER和NCBI数据库对SC124菌株的基因组序列进行分析；用肉汤稀释法测定最小抑菌浓度；通过接合试验验证ICE_Prophage携带耐药基因的可转移性；采用反向PCR检测环状中间体的存在；用竞争试验评估接合子是否具有适应性代价。结果显示：猪链球菌SC124菌株中存在携带四环素类、大环内酯类和氨基糖苷类耐药基因的ICE_Prophage(ICESsuSC124_ΦSsuSC124)；携带四环素类耐药基因的ICESsuSC124以5.52×10^-9频率发生转移，未发现前噬菌体发生转移；ICESsuSC124可以形成一个大小约为3.8 kb的环状中间体；与猪链球菌BAA相比，含有ICESsuSC124的接合子在无药物压力选择的作用下不具有竞争优势，接合子会产生一定程度的适应性代价。综上，猪链球菌SC124菌株中存在携带四环素类、大环内酯类和氨基糖苷类耐药基因的ICE_Prophage，ICESsuSC124可发生水平转移，其接合子可产生一定程度的适应性代价。

关键词: 猪链球菌, ICE_Prophage, 接合试验, 适应性代价

Abstract:

This study aimed to investigate the transfer of antibiotic resistance genes (ARGs) carried by ICE_Prophage in Streptococcus suis. The whole genome sequencing of S. suis SC124 was performed using the Illumina MiSeq and Pacbio RSII platforms. The whole genome sequence of SC124 isolates was analyzed by online websites RAST, ICEberg 3.0, PHASTER and NCBI database. MICs were determined by broth microdilution. The transferability of ICE_Prophage carrying ARGs in S. suis was investigated by conjugation. The present of circular intermediates was examined by inverse PCR. Competition was used to evaluate whether the transconjugant had fitness cost. The results showed that there was an ICE_Prophage (ICESsuSC124_ΦSsuSC124) carrying tetracycline, macrolide and aminoglycoside resistance genes in S. suis SC124. ICESsuSC124 carrying tetracycline resistance genes was transferred at a frequency of 5.52×10^-9, and no prophage transfer was found. Inverse PCR revealed that ICESsuSC124 formed a circular intermediate with a size of about 3.8 kb. Compared with S. suis BAA, the transconjugant containing ICESsuSC124 did not have a competitive advantage under the action of drug-free pressure selection, and the transconjugant can produce a certain degree of fitness cost. In conclusion, the ICE_Prophage carrying tetracycline, macrolide and aminoglycoside resistance genes in S. suis SC124, in which ICESsuSC124 could be horizontally transferred, and the transconjugant could produce a certain degree of fitness cost.

Key words: Streptococcus suis, ICE_Prophage, conjugation, fitness cost

中图分类号:

S852.611

段慧慧, 任仕航, 张宏印, 于瑞, 刘忠虎, 杜向党, 商艳红. 猪链球菌SC124菌株中ICE_Prophage携带耐药基因的水平转移[J]. 畜牧兽医学报, 2024, 55(8): 3699-3705.

Huihui DUAN, Shihang REN, Hongyin ZHANG, Rui YU, Zhonghu LIU, Xiangdang DU, Yanhong SHANG. Horizontal Transfer of Drug Resistance Genes Carried by ICE_Prophage in Streptococcus suis SC124[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3699-3705.

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菌株Strains	att1	att2	att3
TZ080501	catCAC$\underline{\rm{G}}$T$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $tgt	ccaCAC$\underline{\rm{G}}$T$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $cgt	acaCAC$\underline{\rm{A}}$T$\underline{\rm{A}}$GAagttgt
SC070731	catCAC$\underline{\rm{G}}$T$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $cgt	acaCACGT$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $tgt	ccaCAC$\underline{\rm{A}}$T$\underline{\rm{A}}$GAagttgt
SC124	catCAC$\underline{\rm{G}}$T$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $cgt	acaCAC$\underline{\rm{G}}$T$\underline{\rm{G}}$GA$ \underline {{\rm{gtg}}} $tgt	ccaCAC$\underline{\rm{A}}$T$\underline{\rm{A}}$GAagttgt

菌株 Strains	MICs
菌株 Strains	CLI	LIN	ERY	RFP	FUS	TET	CHL	FFC	AMP
SC124	128	256	256	＜1	4	128	2	＜1	＜1
BAA	＜1	＜1	＜1	128	64	＜1	＜1	＜1	＜1
BAA+ICESsuSC124	＜1	＜1	＜1	128	64	128	＜1	＜1	＜1

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