1株羊源伪结核棒状杆菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.03.041

Abstract

Abstract: 【Objective】 The purpose of this experiment was to identify the pathogen of goats suspected to be suffering from pseudotuberculosis in a farm in Yiliang,Yunnan province,and explore its drug resistance and pathogenicity,so as to provide theoretical basis for the diagnosis and treatment of the disease.【Method】 According to the clinical symptoms of the infected goats,the cause of the disease was preliminarily determined,the disease material was collected aseptically,and the pathogenic bacteria were isolated and purified to identify the morphological characteristics,culture characteristics and biochemical characteristics.The 16S rRNA gene sequences of the isolate was amplified by PCR and analyzed by sequencing.The drug resistance and pathogenicity of the isolate were investigated by drug susceptibility test,drug resistance gene test and pathogenicity test in mice.【Result】 The isolate grew well on the blood agar plate,the colonies were yellowish-white and opaque,and did not grow well in the common nutrient agar and the common nutrient broth.Gram staining microscopy showed that the bacteria were spheroidal or rod-shaped and Gram-positive bacteria.The results of biochemical identification showed that glucose and urea tests were positive,while lactose,maltose,mannitol,sucrose,Simon’s citrate,hydrogen sulfide,indigo matrix,nitrate reduction test,methyl red (MR) test and VP test were negative.16S rRNA sequence alignment results showed that，the 16S rRNA gene sequence similarity between the isolated and Corynebacterium pseudotuberculosis in NCBI database was more than 99%,and it was identified as Corynebacterium pseudotuberculosis.Drug sensitivity test results showed that，the isolate showed intermediation to streptomycin,gentamicin and neomycin,and resistance to amtronam,ceftazidime and polymyxins.The results of drug resistance gene detection showed that the isolate carried drug resistance genes Sul2,Sul3,bla_TEM and bla_CTX-M. The results of animal pathogenicity test showed that abdominal and subcutaneous inoculation with 4.4×10⁸ CFU/mL isolated bacteria could cause death of mice,and the mortality rate was 100%.Subcutaneous abscess appeared 96 h after subcutaneous inoculation,and Corynebacterium pseudotuberculosis could be isolated from the pus.【Conclusion】 In this experiment,a strain of Corynebacterium pseudotuberculosis of goats origin was successfully isolated and identified,and the pathogen of the infected goats was identified.The strain was highly pathogenic to mice.The results provided reference for the diagnosis,epidemiological investigation and treatment of Corynebacterium pseudotuberculosis infection in goats.

Key words: goats; Corynebacterium pseudotuberculosis; isolation and identification; drug resistance

CLC Number:

S852.6

LIU Zewu, FAN Yueyuan, YUANJiarui, GESANG Zhuoga, BAN Dan, BAI Weibing, CHAJinlong, SILANG Yuzhen, FU Guowen. Isolation，Identification and Drug Resistance Analysis of a Strain of Corynebacterium pseudotuberculosis from Goats[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1405-1415.

References

[1] BAIRD G，FONTAINE M.Corynebacterium pseudotuberculosis and its role in ovine caseous lymphadenitis[J].Journal of Comparative Pathology,2007,137(4):179-210.
[2] 耿增强.兽医微生物学[M].北京:中国农业出版社,1998. GENG Z Q.Veterinary Microbiology[M].Beijing:China Agriculture Press,1998.(in Chinese)
[3] 杨雪红,郭雪洁,马宁君,等.羊干酪性淋巴结炎研究进展[J].上海畜牧兽医通讯,2008,1:2-3. YANG X H,GUO X J,MA N J,et al.Research progress of feta cheese lymphadenitis [J].Shanghai Journal of Animal Husbandry and Veterinary Medicine,2008,1:2-3.(in Chinese)
[4] 侯宏虎,王志芳,杨增岐.羊伪结核及大肠埃希菌病二联灭活油苗的试制[J].动物医学进展,2007，10:116-118. HOU H H,WANG Z F,YANG Z Q.Trial production of double inactivated oil seedlings for sheep pseudotuberculosis and coliform disease[J].Progress in Veterinary Medicine,2007，10：116-118.(in Chinese)
[5] 赵宏坤,范伟兴,胡敬东，等.羊伪结核病研究进展[J].中国预防兽医学报,2000，3:77-78. ZHAO H K,FAN W X,HU J D,et al.Research progress on pseudotuberculosis in sheep[J].Chinese Journal of Preventive Veterinary Medicine,2000，3：77-78.(in Chinese)
[6] DORELLA F A,PACHECO L G,OLIVEIRA S C,et al. Corynebacterium pseudotuberculosis:Microbiology,biochemical properties,pathogenesis and molecular studies of virulence[J].Veterinary Research,2006，37(2):201-218.
[7] 崔宁,党光辉,鹿萍,等.山羊源伪结核棒状杆菌的分离鉴定及其毒力评价[J].黑龙江畜牧兽医,2024，10:65-70. CUI N,DANG G H,LU P,et al.Isolation and identification of Corynebacterium pseudotuberculosis from goats and its virulence evaluation[J].Heilongjiang Animal Science and Veterinary Medicine,2024，10：65-70.(in Chinese)
[8] 华丽,李欣.羊伪结核棒状杆菌病的诊断与预防[J].河北农业,2023，4:84-85. HUA L,LI X.Diagnosis and prevention of sheep pseudotuberculosis corynebacillosis[J]. Hebei Agriculture,2023，4:84-85.(in Chinese)
[9] 邹安龙.伪结核棒状杆菌灭活苗制备及对小鼠和山羊的免疫效果探究[D].重庆:西南大学,2023. ZOU A L.Preparation of Corynebacterium pseudotuberculosis inactivated vaccine and its immune effect on mice and goats[D].Chongqing:Southwest University,2023.(in Chinese)
[10] 沈荣华,张晓霞,翁紫安.山羊伪结核棒状杆菌病的诊断和防治[J].中国草食动物,2011,31(1):72-73. SHEN R R,ZHANG X X,WENG Z A.Diagnosis and prevention of Corynebacteriosis pseudotuberculosis in goats[J].China Herbivore Science,2011,31(1):72-73.(in Chinese)
[11] 郑国英,马玉馨,李太仙,等.昆明市西山区山羊伪结核棒状杆菌的分离鉴定[J].动物医学进展,2017,38(9):32-37. ZHENG G Y,MA Y X,LI T X,et al.Isolation and identification of Corynebacterium pseudotuberculosis in goats in Xishan dstrict,Kunming city[J]. Progress in Veterinary Medicine,2017,38(9):32-37.(in Chinese)
[12] 马玉馨,李文贵,郑国英,等.山羊伪结核棒状杆菌TaqMan荧光定量PCR检测方法的建立和初步应用[J].畜牧与兽医,2018,50(10):82-87. MA Y X,LI W G,ZHENG G Y,et al.Establishment and preliminary application of TaqMan Real-time PCR method for detection of Corynebacterium pseudotuberculosis in goats[J].Animal Husbandry & Veterinary Medicine,2018,50(10):82-87.(in Chinese)
[13] 王峰.关中奶山羊伪结核棒状杆菌的分离鉴定及毒力基因和耐药性检测[D].杨凌：西北农林科技大学,2019. WANG F.Isolation and identification of Corynebacterium pseudotuberculosis in dairy goats in Guanzhong and detection of virulence genes and drug resistance[D].Yangling：Northwest A&F University,2019.(in Chinese)
[14] 马苏,高艳春.动物源细菌耐药性对动物细菌传染性疾病防治的影响[J].中国兽药杂志,2012,46(2):50-52. MA S,GAO Y C.Effect of antimicrobial resistance of animal-derived bacteria on the prevention and control of animal bacterial infectious diseases[J].Chinese Journal of Veterinary Drug,2012,46(2):50-52.(in Chinese)
[15] 李文均,陈瑛,田新朋,等.《伯杰氏鉴定细菌学手册》和“伯杰氏国际系统微生物学学会”历史回顾、发展现状及未来展望[J].微生物学报,2023,63(5):1714-1723. LI W J,CHEN Y,TIAN X P,et al.Historical review,development status and future prospect of Berger’s Manual of Identification Bacteriology and Berger’s International Society for Systematic Microbiology [J].Acta Microbiologica Sinica,2023,63(5):1714-1723.(in Chinese)
[16] 庞建,刘占英,郝敏,等.革兰氏阳性细菌基因组DNA提取方法的比较及优化[J].微生物学通报,2015,42(12):2482-2486. PANG J,LIU ZHAN Y,HAO M,et al.Comparison and optimization of genomic DNA extraction methods from Gram-positive bacteria[J].Microbiology China,2015,42(12):2482-2486.(in Chinese)
[17] CETINKAYA B,KARAHAN M,ATIL E,et al.Identification of Corynebacterium pseudotuberculosis isolates from sheep and goats by PCR[J].Veterinary Microbiology,2002,88(1):75-83.
[18] 阮紫涵,黄安雄,王秀娟,等.CLSI、EUCAST和中国耐药判定标准概述[J].生物技术通报,2022，38(9):47-58. RUAN Z H,HUANG A X,WANG X J,et al.Overview of CLSI,EUCAST,and susceptibility breakpoints in China [J].Biotechnology Bulletin,2022,38(9):47-58.(in Chinese)
[19] LIAO X P,LIU B T,YANG Q E,et al.Comparison of plasmids coharboring 16S rRNA methylase and extended-spectrum β-lactamase genes among Escherichia coli isolates from pets and poultry[J].Journal of Food Protection,2013，76(12):2018-2023.
[20] DROPPA-ALMEIDA D,VIVAS W L,SILVA K K,et al.Recombinant CP40 from Corynebacterium pseudotuberculosis confers protection in mice after challenge with a virulent strain[J].Vaccine,2016,34(8):1091-1096.
[21] 朱志雄，梁文娟.浅淡山羊伪结核病的防治[J].广西畜牧兽医，2015，1:32-46. ZHU Z X,LIANG W J.Analysis on the prevention and control of pseudotuberculosis in goats[J].Guangxi Journal of Animal Husbandry & Veterinary Medicine ,2015，1:32-46.(in Chinese)
[22] 潘淑惠,王璇,万晴姣,等.山羊伪结核棒状杆菌感染途径及病理变化观察[J].上海畜牧兽医通讯,2007，5:24-25. PAN S H,WANG X,WAN Q J,et al.Observation on the infection route and pathological changes of Corynebacterium pseudotuberculosis in goats[J].Shanghai Journal of Animal Husbandry and Veterinary Medicine,2007，5:24-25.(in Chinese)
[23] 曾泽，韦雷飞，黄秀君，等.一起山羊伪结核棒状杆菌感染的诊断[J].动物医学进展，2014，3:131-133. ZENG Z,WEI L F,HUANG X J,et al.Diagnosis of Corynebacterium pseudotuberculosis infection in goats[J]. Progress in Veterinary Medicine,2014，3:131-133.(in Chinese)
[24] 黄丽丽,林裕胜,江锦秀,等.山羊伪结核棒状杆菌的分离鉴定及耐药性分析[J].福建农业学报,2019,34(8):925-932. HUANG L L,LIN Y S,JIANG J X,et al.Isolation,identification and drug resistance analysis of Corynebacterium pseudotuberculosis in goats[J].Fujian Journal of Agricultural Sciences,2019,34(8):925-932.(in Chinese)
[25] 李国平,陈家祥,王全溪.波尔山羊伪结核棒状杆菌在我省的首次分离鉴定[J].福建畜牧兽医,2003,5:1-2. LI G P,CHEN J X,WANG Q X.The first isolation and identification of Corynebacterium pseudotuberculosis in Boer goat[J].Fujian Journal of Animal Husbandry & Veterinary Medicine,2003,5:1-2.(in Chinese)
[26] 魏宇辰,王斌,白新栋,等.奶山羊不同组织来源干酪性脓肿中伪结核棒状杆菌的分离及毒力基因和耐药性检测[J].中国兽医学报,2023,43(5):945-950. WEI Y C,WANG B,BAI X D,et al.Isolation of Corynebacterium pseudotuberculosis in caseating abscess of different tissue sources in dairy goats and detection of virulence genes and drug resistance[J].Chinese Journal of Veterinary Science,2023,43(5):945-950.(in Chinese)
[27] 吴浩阳,许信刚,任杰,等.山羊伪结核棒状杆菌的分离与鉴定[J].畜牧与兽医,2018,50(9):74-77. WU H Y,XU X G,REN J,et al.Isolation and identification of Corynebacterium pseudotuberculosis in goats[J].Animal Husbandry & Veterinary Medicine,2018,50(9):74-77.(in Chinese)
[28] RODRÍGUEZ-DOMÍNGUEZ M C,MONTES-DE-OCA-JIMÉNEZ R,VÁZQUEZ-CHAGOYÁN J C,et al.Bioinformatic approach of B and T cell epitopes of PLD and CP40 proteins of Corynebacterium pseudotuberculosis ovis mexican isolate 2J-L towards a peptide-based vaccine[J].International Journal of Molecular Sciences,2023,25(1):270.

Isolation，Identification and Drug Resistance Analysis of a Strain of Corynebacterium pseudotuberculosis from Goats

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	RU Mengke, LI Suixiang, WU Xueqin, YAN Yuzhang, WANG Lu, CHENG Haipeng. Whole Genome Sequencing and Bioinformatics Analysis of a Strain of Proteus mirabilis Isolated from Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 977-989.
[2]	AN Zhaoxiang, XUN Wenjuan, ZHOU Hanlin, SHI Liguang. Effects of Heat Stress on Semen Antioxidant Capacity and Seminal Plasma Metabolomics in Hainan Black Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1191-1201.
[3]	GAO Jiaojiao, ZHENG Nan, SHAO Wei, CHEN He, MA Xianlan, ZHAO Yankun. Isolation and Identification of Lactogenic Streptococcus agalactiae and Characterisation of Drug Resistance and Virulence and Genome Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1301-1316.
[4]	LI Na, LIU Chongyang, ZHANG Jingjing, MALIYA Qiqige, ZHU Na, LU Bin, HAI Ying. Isolation,Identification and Drug Resistance Analysis of a Sheep-derived Strain of Clostridium perfringens Type D [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1352-1359.
[5]	LI Yang, XU Jingjing, ZHANG Xiaoyu, LI Nana, YU Xingyu, LENG Qingwen, LI Yanfang, QU Yonggang. Isolation,Identification,Drug Resistance Analysis,and Virulence Gene Detection of Staphylococcus aureus from a Large-scale Dairy Farm in Xinjiang [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1370-1382.
[6]	DAI Tingting, CHEN Haiyu, DUAN Chuchu, LIU Rongchang, LI Yurong, YAN Shuhan, CHEN Mengshi, LIU Luwei, BAO Yinli, CHENG Yanqing, LIN Weiming, HUANG Cuiqin, ZHENG Xintian. Correlation Analysis of Drug Resistance and Virulence Genes of Escherichia coli with Polycolistin Resistance Gene mcr-1 from Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1393-1404.
[7]	AI Xiaonan, CHENG Lifen, BAI Pu, CHEN Zhenghao, XUE Lina, ZHOU Shenghua. Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 499-511.
[8]	LEI Zhigang, PAN Hong, ZHANG Dandan, LIU Quanhui, SUN Zhe, DENG Shan, HUANG Ben. Cloning, Bioinformatics Analysis and Eukaryotic Expression Vector Construction of MAPK8 Gene in Mashan Black Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 534-544.
[9]	LI Jingxuan, LIN Yanjiao, HUANG Qiongjun, HAN Xinyan, ZHANG Yuelang. Research Progress on Non-coding RNA Related to Skeletal Muscle Development in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 582-592.
[10]	LI Qianwen, HUA Tao, CHANG Chen, YAN Xingyu, LI Junxing, TANG Bo. Isolation,Identification and Pathogenicity Analysis of a Strain of Haemophilus parasuis Serotype 2 [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 901-911.
[11]	WAN Baoxia, MENG Lingying, SUN Siyu, ZHAO Yujie, WANG Jiaqi, WANG Qiuju. Analysis of Drug Resistance Genes and Biological Characterization of Goose Derived Bacillus and Lactic Acid Bacteria [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 351-363.
[12]	LI Nana, YU Xingyu, HOU Gongmingzhu, LI Yang, GUO Yaqi, ZHENG Pei, LI Yanfang, LIANG Yan, HE Gaoming, QU Yonggang. Isolation and Identification,Drug Resistance and Pathogenicity Analysis of Enterobacter hormaechei from Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 376-388.
[13]	HUANG Yao, YAO Naiquan, YUAN Weitao, MAO Aipeng, ZHANG Ting, XU Chao. Isolation and Identification of Candida rugosa DZ018 from Deer and Its Probiotic Properties in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 389-399.
[14]	LIAO Huiqun, ZHAO Mei, ZENG Guohui, SU Renwei, DENG Xianbo. Analysis of Drug Resistance and Virulence Genes of Klebsiella pneumoniae from Moschus berezovskii [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 411-421.
[15]	JIANG Jiang, NIAN Linyu, CHEN Lin, LIN Chen, CAO Chongjiang. Isolation,Identification and Probiotic Characteristics of Bacillus siamensis in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 432-441.