猪源致病性格氏乳球菌的分离鉴定及毒力基因分析

doi:10.11843/j.issn.0366-6964.2025.07.032

摘要/Abstract

摘要：

本研究旨在研究猪源格氏乳球菌的生物学特征、药物敏感性及致病性。从患病猪肝脏中分离到可疑细菌，并对分离的细菌进行革兰染色镜检、基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)鉴定、16S rRNA基因序列测定及序列分析；通过药敏试验和小鼠感染试验探究分离菌株的耐药性和致病性；通过PCR扩增筛选分离菌株相关毒力基因。结果显示：从患病猪肝脏中分离到1株产α-溶血环的阳性球菌；MALDI-TOF MS分析、16S rRNA基因测序及进化树分析均显示分离菌株为格氏乳球菌；小鼠感染试验显示，分离菌株对小鼠有较强的致死性，能引起小鼠肝、脾肿大，肠道、腹腔、胸腔严重充血；药敏试验结果显示，分离菌株对β-内酰胺类药物如青霉素G、氨苄西林，以及红霉素、林可霉素耐药，对其它抗生素均敏感；毒力基因检测结果显示，分离菌株含有黏附相关基因adhC1、psaA、eno以及lp1，含有溶血相关基因hly1、hly2、hly3，含有荚膜基因簇形成相关基因cgcC，含有外多糖相关基因epsA、epsB、epsC、epsD、epsL，以及其他毒力基因nadho、sod、pgm、chp。本试验分离的猪格氏乳球菌具有较强的致病性，因其属于人畜共患病原体，带来的潜在风险不可忽视，本研究为临床格氏乳球菌感染的防治提供数据参考。

关键词: 格氏乳球菌, 猪, 16S rRNA, 药敏试验, 毒力基因

Abstract:

This study aimed to explore the biological characteristics, drug sensitivity and pathogenicity of Lactococcus garvieae from pig. Suspicious bacteria were isolated from the liver of diseased pig, and then were identified by Gram stained microscopy, matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), 16S rRNA gene sequence analysis and sequence analysis. The drug resistance and pathogenicity of isolated strains were explored through drug sensitivity testing and mice infection testing. Related virulence of isolated strain genes were amplified by PCR. Results were as follows: A positive strain of α-hemolytic was isolated from the liver of a diseased pig. MALDI-TOF MS, PCR amplification of 16S rRNA gene and phylogenetic tree analysis results showed that the isolated strain was identified as Lactococcus garvieae.The infection test of mice showed that the isolated strain had strong lethality. It can cause swelling of the liver and spleen, severe congestion of the intestines, abdominal cavity, and chest cavity in mice. The drug sensitivity test results showed that the isolated strains were resistant to β-lactam drugs such as penicillin G and ampicillin, as well as erythromycin and lincomycin, and sensitive to other antibiotics. The results of virulence gene testing showed that the isolated strain contains adhesion related genes adhC1, psaA, eno, lp1, hemolysis related genes hly1, hly2, hly3, capsule gene cluster formation related genes cgcC, polysaccharide related genes epsA, epsB, epsC, epsD, epsL, and other virulence genes nadho, sod, pgm, chp. The Lactococcus garvieae was isolated in this experiment has strong pathogenicity, and its potential risks cannot be ignored as it belongs to zoonotic pathogens. This study provides data reference for the prevention and treatment of clinical infections caused by Lactococcus garvieae.

Key words: Lactococcus garvieae, pig, 16S rRNA, drug susceptibility test, virulence gene

中图分类号:

S858.285.1

舒静超, 张菡, 彭志锋, 乔宏兴. 猪源致病性格氏乳球菌的分离鉴定及毒力基因分析[J]. 畜牧兽医学报, 2025, 56(7): 3399-3407.

SHU Jingchao, ZHANG Han, PENG Zhifeng, QIAO Hongxing. Isolation, Identification and Virulence Gene Analysis of Porcine Pathogenic Lactococcus garvieae[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3399-3407.

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抗菌药物Antimicrobials	抑菌圈直径折点(D)/mm Zone diameter breakpoints			抑菌圈直径/mmZone diameter	敏感性Sensitivity
抗菌药物Antimicrobials	R	I	S	抑菌圈直径/mmZone diameter	敏感性Sensitivity
大观霉素Spectinomycin	≤14	14＜D＜18	≥18	19	S
庆大霉素Gentamicin	≤12	12＜D＜15	≥15	25	S
四环素Tetracycline	≤4	4＜D＜19	≥19	22	S
多西环素Doxycycline	≤9	9＜D＜13	≥13	27	S
头孢唑林Cefuroxime	≤19	19＜D＜23	≥23	31	S
头孢他啶Ceftazidime	≤17	17＜D＜21	≥21	30	S
头孢吡肟Cefepime	≤18	18＜D＜25	≥25	33	S
氧氟沙星Ofloxacin	≤13	13＜D＜17	≥17	22	S
环丙沙星Ciprofloxacin	≤15	15＜D＜21	≥21	29	S
恩诺沙星Enrofloxacin	≤15	15＜D＜21	≥21	22	S
阿米卡星Amikacin	≤14	14＜D＜17	≥17	18	S
加替沙星Gatifloxacin	≤14	14＜D＜18	≥18	23	S
青霉素G Penicillin G	≤28	-	≥29	0	R
氨苄西林Ampicillin	≤13	13＜D＜17	≥17	0	R
红霉素Erythromycin	≤13	13＜D＜23	≥23	9	R
氯霉素Chloramphenicol	≤12	12＜D＜18	≥18	22	S
新霉素Neomycin	≤16	16＜D＜22	≥22	30	S
阿奇霉素Azithromycin	≤13	13＜D＜18	≥18	15	I
复方新诺明Bactrim	≤10	10＜D＜16	≥16	37	S
磺胺异恶唑Sulfisoxazole	≤12	12＜D＜17	≥17	21	S
亚胺培南Imipenem	≤19	19＜D＜23	≥23	15	R
美罗培南Meropenem	≤19	19＜D＜23	≥23	24	S
林可霉素Lincomycin	≤9	9＜D＜15	≥15	0	R
多黏菌素B Polymyxin B	≤8	8＜D＜12	≥12	25	S

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