鸭源大肠杆菌的分离鉴定及致病性分析

doi:10.16431/j.cnki.1671-7236.2022.05.039

Abstract

Abstract: 【Objective】 This study was aimed to understand the distribution and pathogenicity of Escherichia coli from ducks in Jiangsu,Jiangxi and Anhui provinces.【Method】 This study isolated and identified duck derived Escherichia coli from dead ducks in Jiangsu,Jiangxi and Anhui,determined the serotype of duck derived Escherichia coli isolates by PCR combined with slide agglutination,detected 18 virulence genes by PCR,and then carried out the pathogenicity test of ducklings.The growth curve and median lethal dose (LD₅₀) of the strains with strong and weak virulence were determined.【Result】 A total of 74 strains of duck derived Escherichia coli were isolated and identified in this study,1,2,2 and 4 strains were identified as O1,O2,O18 and O78 serotypes respectively,and the rest were not finalized.The identification test of 18 virulence genes showed that positive rates of genes ibeB,yijp,ompA and mat were 97.3%,97.3%,95.95% and 90.54%,respectively.Animal pathogenicity tests showed that 74 strains of Escherichia coli isolates were pathogenic to ducklings at infection dose of 10⁷ CFU/duck,but only 2 strains of isolates caused the death rate of ducklings ≥50%.The growth curve detemination showed that there was no significant difference between 2 strains of virulent isolates and 2 strains of attenuated isolates (P>0.05),and the LD₅₀ of 2 strains of virulent isolates were 10^4.75and 10^7.375 CFU,respectively.【Conclusion】 The rate of serotypes O1,O2,O18,O78 in 74 isolates was only 12.16%.The virulence gene spectrum was widely distributed,but only 2 strains of isolates were more virulent.This study laid the foundation for the prevention and control of Escherichia coli from ducks,it was also helpful for studying the relationship among serotypes,virulence genes and pathogenicity.

Key words: Escherichia coli from ducks; serotypes; virulence genes; animal pathogenicity test

CLC Number:

S852.61

DONG Hongyan, SONG Haigang, ZHU Shanyuan, YU Shengqing, XIE Jun, HONG Shuangpeng, GUO Changming, XU Hai, GAO Yuexiu. Isolation,Identification and Pathogenicity Analysis of Escherichia coli from Ducks[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1977-1984.

References

[1] 杨立军,韩先干,尹会方,等.福建省龙岩市禽致病性大肠杆菌的血清型、毒力因子及耐药研究[J].中国动物传染病学报,2016,24(6):24-29. YANG L J,HAN X G,YIN H F,et al.Serotypes,pathogenicity and antibiotic resistance of avian pathogenic Escherichia coli strains from Longyan city,Fujian province[J].Chinese Journal of Animal Infectious Diseases,2016,24(6):24-29.(in Chinese)
[2] 胡紫萌,王卓昊,吴坤,等.苏北及周边地区水禽致病性大肠杆菌的分离鉴定及耐药性分析[J].中国预防兽医学报,2020,42(5):439-444. HU Z M,WANG Z H,WU K,et al.Isolation,identification and drug resistance analysis of waterfowl avian pathogenic Escherichia coli (APEC) in Northern Jiangsu and surrounding areas[J].Chinese Journal of Preventive Veterinary Medicine,2020,42(5):439-444.(in Chinese)
[3] 李迎晓,焦凤超,张李敏,等.鸭源大肠杆菌分离鉴定及其毒力基因检测[J].畜牧与兽医,2020,52(3):73-77. LI Y X,JIAO F C,ZHANG L M,et al.Isolation of duck-derived pathogenic Escherichia coli and test of virulence genes[J].Journal of Animal Husbandry & Veterinary Medicine,2020,52(3):73-77.(in Chinese)
[4] 孟庆美,王少辉,韩先干,等.禽致病性大肠杆菌毒力基因多重PCR方法的建立和应用[J].微生物学报,2014,54(6):696-702. MENG Q M,WANG S H,HAN X G,et al.Multiplex PCR assay for detection of virulence genes in avian pathogenic Escherichia coli[J].Acta Microbiologica Sinica,2014,54(6):696-702.(in Chinese)
[5] 孙畅,牛银杰,尹海畅,等.鸭致病性大肠杆菌研究进展[J].黑龙江畜牧兽医,2017,8:72-77. SUN C,NIU Y J,YIN H C,et al.Research progress on avian pathogenic Escherichia coli of duck[J].Heilongjiang Animal Science and Veterinary Medicine,2017,8:72-77.(in Chinese)
[6] WANG S H,MENG Q M,DAI J J,et al.Development of an allele-specific PCR assay forsimultaneous sero-typing of avian pathogenic Escherichia coli predominant O1,O2,O18 and O78 strains[J].PLoS One,2014,9(5):e96904.
[7] EWERS C,JANSSEN T,KIESSLING S,et al.Molecular epidemiology of avian pathogenic Escherichia coli(APEC) isolated from coli septicemia in poultry[J].Veterinary Microbiology,2004,104(1/2):91-101.
[8] RITCHIE J M,THORPE C M,ROGERS A B,et al.Critical roles for stx2,eae,and tir in enter ohemorrhagic Escherichia coli-induced diarrhea and intestinal inflammation in infant rabbits[J].Infection Immunity,2003,71(1):7129-7139.
[9] LINDSTEDT B A,HEIR E,NYGARD I,et al.Characterisation of class Ⅰ integrons in clinical strains of Salmonella enterica subsp.enterica serovars typhimurium and Enteritidis from Norwegian hospitals[J].Joural of Medical Microbiology,2003,52(2):141-149.
[10] REED L J,MUENCH H.A simple method of estimating fifty percent endpoints[J].American Journal of Hygiene,1938,27:493-497.
[11] 沈达,王健,宋海港,等.水禽源致病性大肠杆菌流行病学调查及耐药性分析[J].现代畜牧兽医,2021,1:67-70. SHEN D,WANG J,SONG H G,et al.Epidemiological investigation and drug resistance analysis of pathogenic waterfowl source E.coli[J].Modern Journal of Animal Husbandry and Veterinary Medicine,2021,1:67-70.(in Chinese)
[12] 董向磊.禽致病性大肠杆菌的分离鉴定和分离株毒力基因与致病性相关性研究[D].扬州:扬州大学,2014. DONG X L.Isolation and identification of avian pathogenic Escherichia coli and the correlation of AEPC virulence genes with its pathogenicity[D].Yangzhou:Yangzhou University,2014.(in Chinese)
[13] 于学辉,程安春,王远微,等.规模化养鸭场鸭源大肠杆菌分离株的致病性及系统发育分析[J].中国预防兽医学报,2011,33(12):927-931. YU X H,CHENG A C,WANG Y W,et al.Pathogenicity and phylogenetic analysis of duck pathogenic E.coli from large-scale duck farms[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(12):927-931.(in Chinese)
[14] 吴信明,连洪梅,王艳,等.38株鸭源大肠杆菌的分离鉴定与药敏试验[J].中国畜牧兽医,2010,37(10):178-180. WU X M,LIAN H M,WANG Y,et al.Isolation,identification and drug sensitivity test of 38 strains of Escherichia coli from duck[J].China Animal Husbandry & Veterinary Medicine,2010,37(10):178-180.(in Chinese)
[15] 张济培,韦庆兰,谭华龙,等.广东地区水禽大肠杆菌血清型鉴定[J].中国兽医杂志,2015,51(8):41-43. ZHANG J P,WEI Q L,TAN H L,et al.Serotype identification of Escherichia coli from waterfowl in Guangdong[J].Chinese Journal of Veterinary Medicine,2015,51(8):41-43.(in Chinese)
[16] CROXEN M A,FINLAY B B.Molecular mechanisms of Escherichia coli pathogenicity[J].Nature Review Microbiology,2010,8(1):26-38.
[17] 师震宇.禽致病性大肠杆菌鸭源分离株侵袭相关基因致病机理研究[D].南京:南京农业大学,2011. SHI Z Y.Pathogenic mechanism of invasion associated genes in avain pathogenic Escherichia coli duck isolate[D].Nanjing:Nanjing Agricultural University,2011.(in Chinese)
[18] 贾青辉,张召兴,李蕴玉,等.河北地区鸡源致病性大肠杆菌毒力基因检测与耐药性分析[J].中国预防兽医学报,2018,40(5):401-405. JIA Q H,ZHANG Z X,LI W Y,et al.Analys is of virulence genes and drug resistance of the pathogenic Escherichia coli isolates from chicken in Hebei province[J].Chinese Journal of Preventive Veterinary Medicine,2018,40(5):401-405.(in Chinese)
[19] EWERS C,JANSSEN T,KIESSLING S,et al.Rapid detection of virulence-associated genes in avian pathogenic Escherichia coli by multiplex polymerase chain reaction[J].Avian Diseases,2005,49:269-273.
[20] 王春莲,陈志军,魏晓巍,等.承德滦平地区鹅致病性大肠杆菌的分离鉴定与药敏试验分析[J].黑龙江畜牧兽医,2017,10:126-128. WANG C L,CHEN Z J,WEI X W,et al.Isolation,identification and drug sensitivity analysis of pathogenic Escherichia coli from geese in Luanping area of Chengde[J].Heilongjiang Animal Science and Veterinary Medicine,2017,10:126-128.(in Chinese)

Isolation,Identification and Pathogenicity Analysis of Escherichia coli from Ducks

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CAO Qi, JIN Qing, YUAN Yi, FENG Yixue, KONG Danni, CHEN Xianghong, WU Bin, SONG Wenbo, TANG Xibiao. Epidemiological Investigation of Common Respiratory Bacterial Pathogens in Large-scale Pig Farms [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2439-2449.
[2]	WANG Zhifang, XU Yindi, ZHU Wenhao, ZHANG Qingxian, JIAO Wenqiang, LI Haili, WANG Keling. Analysis of Virulence and Drug Resistance of Haemophilus parasuis Serotype 7 Strains [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3245-3255.
[3]	MAI Zhanhai, LU Yabin, LI Jianlong, ZHAO Lu, SUN Danlan, TONG Panpan, KUANG Ling. Effects of Honeysuckle, Forsythia and Their Drug Pair on Resistance Genes and Virulence Genes of Streptococcus equi subsp. equi [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 709-717.
[4]	JIA Tong, ZHANG Di, ZHANG Zhaotian, CHEN Panliang, ZHANG Jie, LI Yinghui, ZHU Yue, QIN Jianhua, LI Yan. Detection of Virulence and Resistance Genes and Drug Sensitivity Analysis of Escherichia coli Derived from Diarrheic Calves in Beijing-Tianjin-Hebei Partial Regions [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4820-4831.
[5]	ZHANG Haolei, SALANG Wenzhu, HUANG Zhihong, WEN Yongping, ZHANG Huanrong. Isolation, Identification, Drug Resistance and Pathogenicity Analysis of Salmonella from Duck [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4887-4897.
[6]	ZHANG Su, FANG Yuwen, CHEN Yunsheng, GAO Li, XU Guiyun, ZHU Kui, ZHENG Shijun, WANG Yongqiang. Isolation,Identification and Antibiotic Resistance Analysis of Bacillus cereus on Eggshell [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4476-4484.
[7]	QU Zhehui, ZHANG Xiwen, ZHAO Yu, ZHENG Quanfang, LIAN Huixiang, ZHAO Yunhuan, GUO Xiaoqiu. Genetic Variation Analysis of Major Virulence Genes of Pseudorabies Virus Epidemic Strains in Southern Henan Province [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 3019-3029.
[8]	GE Qiang, MA Dongxin, ZHOU Yuqing, CHEN Huaijun, YUAN Jingzhi, LI Xun, WANG Xiaoye. Isolation,Identification and Biological Characteristics of Proteus mirabilis of Swine [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1804-1815.
[9]	DENG Zhidan, HUANG Liangzong, BAI Aiquan. Isolation,Identification and Biological Characteristics of Streptococcus suis Serotype 2 Strain from Guangdong Province [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 717-725.
[10]	WU Zhengzhuo, CHEN Guoquan, YAO Biqiong, SHI Dajun, YAN Chaohua, WANG Na, WANG Kaigong, ZHOU Bijun, CHENG Zhentao, WEN Ming. Isolation, Identification, Virus Gene Detection and Antibiotic Resistance Analysis of Riemerella anatipestifer in Sansui County of Guizhou Province [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4274-4283.
[11]	CHEN Yaqiang, PENG Jinjin, LIAO Ming, LIAO Qinfeng. Virulence Genes Detection and Antibiotic Resistance Analysis of Escherichia coli Isolated from Slaughtering Process of Free-range Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2615-2624.
[12]	WANG Dong, JIN Shengzi, DU Yuanyi, GUAN Tongxu, LI Wanying, LIU Yun. Screening of Sensitive Drugs for Bovine Pathogenic Escherichia coli and Bacteriostatic Test of Sodium Humate [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3361-3370.
[13]	WANG Fei, WANG Cailian, SONG Shuzhen, LANG Xia. Main Pathogenic Bacteria Causing Subclinical Mastitis in Yak and the Distribution of Antibiotic Resistance and Virulence Genes [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 229-239.
[14]	HAO Dongmin, ZHONG Hongyi, CHENG Binghua, LU Ailing, XU Chuanbao, MA Youquan. Diagnosis of Riemerella anatipestifer Infection and Identification of the Pathogens [J]. , 2019, 46(4): 1174-1180.
[15]	LIU Lijuan, MA Guangqiang, ZHANG Shengzhan, WANG Wei, WANG Kaigong, ZHOU Bijun, GOU Wanli. Classification and Drug Resistance Analysis of Pathogenic Escherichia coli in Diarrhea Piglets [J]. , 2018, 45(10): 2876-2884.