A:L1 ST128型鸭源多杀性巴氏杆菌的耐药性及毒力分析

doi:10.11843/j.issn.0366-6964.2021.010.016

Abstract

Abstract: The aim of this study was to characterize the drug resistance and virulence of pathogens from ducks suspected of cholera in a farm in Guizhou. The pathogenic bacteria were cultured and confirmed by PCR. Capsule genotyping and lipopolysaccharide genotyping, multilocus sequence typing (MLST), ERIC-PCR, animal regression test, drug susceptibility test, whole genome sequencing(WGS) and comparative analysis of local genomes were conducted on those isolated strains. Five Pasteurella multocida isolates (PmCW1-5) were identified as A:L1 ST128 and shared high homology. All of them were resistant to ampicillin, amoxicillin, levofloxacin, and lincomycin. PmCW1 also showed low level resistance to ciprofloxacin. WGS data showed PmCW1 which is highly virulent also harbored genes mediating β-lactams, quinolones, tetracyclines, macrolides and peptides resistance and encoding multi-drug efflux pumps and multi-drug resistance proteins. Meanwhile, PmCW1 carried 201 different virulence-mediating genes including those encoding lipooligosaccharide/lipopolysaccharide (LOS/LPS), capsule, adhesion factor, and etc. In addition, type IV fimbriae genes (ptfA, comE, hofB, hofC, vfr), iron uptake related protein genes (ccmABCEF, hgbBC, fur, hscB), outer membrane protein genes (ompP5) were also detected. PmCW1 has the characteristics of typical A:L1 P. multocida. P. multocida ST128 isolated from ducks in this study has been rarely reported. The drug resistance and virulence analysis on those isolates will provide theoretical basis for clinical medication and vaccine development of duck cholera.

Key words: Pasteurella multocida from ducks, isolation and identification, MLST, drug resistance and virulence, comparative analysis of local genomes

CLC Number:

S852.61

ZHANG Yanan, LI Yafei, CHEN Rujia, YU Bo, PENG Shan, LI Ting, PU Ling, XU Jinge. Resistance and Virulence Analysis of Type A: L1 ST128 Pasteurella multocida from Ducks[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2852-2863.

References 44

[1]	WILKIE I W, HARPER M, BOYCE J D, et al. Pasteurella multocida:diseases and pathogenesis[M]//AKTORIES K, ORTH J H C, ADLER B. Pasteurella Multocida. Berlin:Springer, 2012:1-22.
[2]	WILSON B A, HO M. Pasteurella multocida:from zoonosis to cellular microbiology[J]. Clin Microbiol Rev, 2013, 26(3):631-655.
[3]	PENG Z, LIANG W, WANG F, et al. Genetic and phylogenetic characteristics of Pasteurella multocida isolates from different host species[J]. Front Microbiol, 2018, 9:1408.
[4]	HONNORAT E, SENG P, SAVINI H, et al. Prosthetic joint infection caused by Pasteurella multocida:a case series and review of literature[J]. BMC Infect Dis, 2016, 16(1):435.
[5]	RYAN J M, FEDER JR H M. Dog licks baby. Baby gets Pasteurella multocida meningitis[J]. Lancet, 2019, 393:E41.
[6]	LIU Q, HU Y L, LI P, et al. Identification of fur in Pasteurella multocida and the potential of its mutant as an attenuated live vaccine[J]. Front Vet Sci, 2019, 6:5.
[7]	HOLSCHBACH C L, AULIK N, POULSEN K, et al. Prevalence and temporal trends in antimicrobial resistance of bovine respiratory disease pathogen isolates submitted to the Wisconsin Veterinary Diagnostic Laboratory:2008-2017[J]. J Dairy Sci, 2020, 103(10):9464-9472.
[8]	刘海燕, 杨有文, 王远微, 等. 6株山羊源多杀性巴氏杆菌血清型、耐药性及致病性分析[J]. 黑龙江畜牧兽医, 2020, (16):65-68.LIU H Y, YANG Y W, WANG W Y, et al. Analysis of serotype, drug resistance and pathogenicity of 6 strains of Pasteurella multocida from goats[J]. Heilongjiang Animal Science and Veterinary Medicine, 2020, (16):65-68. (in Chinese)
[9]	PRAJAPATI A, CHANDA M M, DHAYALAN A, et al. Variability in in vitro biofilm production and antimicrobial sensitivity pattern among Pasteurella multocida strains[J]. Biofouling, 2020, 36(8):938-950.
[10]	DABO S M, TAYLOR J D, CONFER A W. Pasteurella multocida and bovine respiratory disease[J]. Anim Health Res Rev, 2007, 8(2):129-150.
[11]	HARPER M, BOYCE J D, ADLER B. Pasteurella multocida pathogenesis:125 years after pasteur[J]. FEMS Microbiol Lett, 2006, 265(1):1-10.
[12]	TOWNSEND K M, BOYCE J D, CHUNG J Y, et al. Genetic organization of Pasteurella multocida cap loci and development of a multiplex capsular PCR typing system[J]. J Clin Microbiol, 2001, 39(3):924-929.
[13]	PENG Z, LIANG W, LIU W J, et al. Genomic characterization of Pasteurella multocida HB01, a serotype A bovine isolate from China[J]. Gene, 2016, 581(1):85-93.
[14]	MARZA A D, JESSE ABDULLAH F F, AHMED I M, et al. The ability of lipopolysaccharide (LPS) of Pasteurella multocida B:2 to induce clinical and pathological lesions in the nervous system of buffalo calves following experimental inoculation[J]. Microb Pathog, 2017, 104:340-347.
[15]	PERIASAMY S, PRAVEENA P E, SINGH N. Effects of Pasteurella multocida lipopolysaccharides on bovine leukocytes[J]. Microb Pathog, 2018, 119:225-232.
[16]	HARPER M, JOHN M, TURNI C, et al. Development of a rapid multiplex PCR assay to genotype Pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus[J]. J Clin Microbiol, 2015, 53(2):477-485.
[17]	PENG Z, WANG X R, ZHOU R, et al. Pasteurella multocida:genotypes and genomics[J]. Microbiol Mol Biol Rev, 2019, 83(4):e00014-19.
[18]	Clinical Laboratory Standards Institute (CLSI). Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals;Approved standard-fourth edition. CLSI document VET01-A4[R]. Wayne, PA, USA:CLSI, 2013.
[19]	WICK R R, JUDD L M, GORRIE C L, et al. Unicycler:resolving bacterial genome assemblies from short and long sequencing reads[J]. PLoS Comput Biol, 2017, 13(6):e1005595.
[20]	BESEMER J, LOMSADZE A, BORODOVSKY M. GeneMarkS:a self-training method for prediction of gene starts in microbial genomes. Implications for finding sequence motifs in regulatory regions[J]. Nucleic Acids Res, 2001, 29(12):2607-2618.
[21]	BUCHFINK B, XIE C, HUSON D H. Fast and sensitive protein alignment using DIAMOND[J]. Nat Methods, 2015, 12(1):59-60.
[22]	SULLIVAN M J, PETTY N K, BEATSON S A. Easyfig:a genome comparison visualizer[J]. Bioinformatics, 2011, 27(7):1009-1010.
[23]	刘玉庆, 李璐璐, 骆延波, 等. EUCAST 2017欧盟药敏试验标准[M]. 北京:中国标准出版社, 2016.LIU Y Q, LI L L, LUO Y B, et al. European committee on antimicrobial susceptibility testing[M]. Beijing:Standards Press of China, 2016. (in Chinese)
[24]	王林柏, 孙久鹤, 郭东春, 等. 国内部分地区多杀性巴氏杆菌荚膜血清型和基因型的研究[J]. 中国预防兽医学报, 2016, 38(2):116-119.WANG L B, SUN J H, GUO D C, et al. Identification of capsule serotype and genotype of Pasteurella multocida in some areas of China[J]. Chinese Journal of Preventive Veterinary Medicine, 2016, 38(2):116-119. (in Chinese)
[25]	LI Z C, CHENG F J, LAN S M, et al. Investigation of genetic diversity and epidemiological characteristics of Pasteurella multocida isolates from poultry in southwest China by population structure, multi-locus sequence typing and virulence-associated gene profile analysis[J]. J Vet Med Sci, 2018, 80(6):921-929.
[26]	UJVÁRI B, MAKRAI L, MAGYAR T. Virulence gene profiling and ompA sequence analysis of Pasteurella multocida and their correlation with host species[J]. Vet Microbiol, 2019, 233:190-195.
[27]	EWERS C, LUBKE-BECKER A, BETHE A, et al. Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status[J]. Vet Microbiol, 2006, 114(3-4):304-317.
[28]	TURNI C, SINGH R, BLACKALL P J. Genotypic diversity of Pasteurella multocida isolates from pigs and poultry in Australia[J]. Aust Vet J, 2018, 96(10):390-394.
[29]	UJVÁRI B, WEICZNER R, DEIM Z, et al. Characterization of Pasteurella multocida strains isolated from human infections[J]. Comp Immunol, Microbiol Infect Dis, 2019, 63:37-43.
[30]	PETERSEN A, BISGAARD M, TOWNSEND K, et al. MLST typing of Pasteurella multocida associated with haemorrhagic septicaemia and development of a real-time PCR specific for haemorrhagic septicaemia associated isolates[J]. Vet Microbiol, 2014, 170(3-4):335-341.
[31]	WANG Y H, ZHU J, LU C P, et al. Evidence of circulation of an epidemic strain of Pasteurella multocida in Jiangsu, China by multi-locus sequence typing (MLST)[J]. Infect, Genet Evol, 2013, 20:34-38.
[32]	李伟杰, 田野, 岂晓鑫, 等. 禽源多杀性巴氏杆菌多位点序列分型研究[J]. 中国兽药杂志, 2017, 51(2):1-6.LI W J, TIAN Y, QI X X, et al. Multi-locus sequence typing of Pasteurella multocida isolated from Avian[J]. Chinese Journal of Veterinary Drug, 2017, 51(2):1-6. (in Chinese)
[33]	陈红梅, 程龙飞, 邓辉, 等. 鸭源多杀性巴氏杆菌对磺胺类药物的耐药性检测及耐药基因分析[J]. 中国畜牧兽医, 2019, 46(1):223-230.CHEN H M, CHENG L F, DENG H, et al. Detection of sulfonamides sensitivity and analysis of resistant genes of Pasteurella multocida isolated from ducks[J]. China Animal Husbandry & Veterinary Medicine, 2019, 46(1):223-230. (in Chinese)
[34]	陈国权, 张旭, 阎朝华, 等. 鸭源多杀性巴氏杆菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2020, 47(2):620-628.CHEN G Q, ZHANG X, YAN C H, et al. Isolation, identification and drug resistance analysis of Pasteurella multocida from ducks[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(2):620-628. (in Chinese)
[35]	HOOPER D C, JACOBY G A. Mechanisms of drug resistance:quinolone resistance[J]. Ann N Y Acad Sci, 2015, 1354(1):12-31.
[36]	KONG L C, GAO D, GAO Y H, et al. Fluoroquinolone resistance mechanism of clinical isolates and selected mutants of Pasteurella multocida from bovine respiratory disease in China[J]. J Vet Med Sci, 2014, 76(12):1655-1657.
[37]	HAN J, SAHIN O, BARTON Y W, et al. Key role of Mfd in the development of Fluoroquinolone resistance in Campylobacter jejuni[J]. PLoS Pathog, 2008, 4(6):e1000083.
[38]	RUFFOLO C G, TENNENT J M, MICHALSKI W P, et al. Identification, purification, and characterization of the type 4 fimbriae of Pasteurella multocida[J]. Infect Immun, 1997, 65(1):339-343.
[39]	HATFALUDI T, AL-HASANI K, BOYCE J D, et al. Outer membrane proteins of Pasteurella multocida[J]. Vet Microbiol, 2010, 144(1-2):1-17.
[40]	ALLEN J W A, LEACH N, FERGUSON S J. The histidine of the c-type cytochrome CXXCH haem-binding motif is essential for haem attachment by the Escherichia coli cytochrome c maturation (Ccm) apparatus[J]. Biochem J, 2005, 389:587-592.
[41]	JACOBSEN I, GERSTENBERGER J, GRUBER A D, et al. Deletion of the ferric uptake regulator Fur impairs the in vitro growth and virulence of Actinobacillus pleuropneumoniae[J]. Infect Immun, 2005, 73(6):3740-3744.
[42]	HARPER M, COX A D, ADLER B, et al. Pasteurella multocida lipopolysaccharide:the long and the short of it[J]. Vet Microbiol, 2011, 153(1-2):109-115.
[43]	PETRUZZI B, BRIGGS R E, SWORDS W E, et al. Capsular polysaccharide interferes with biofilm formation by Pasteurella multocida Serogroup A[J]. mBio, 2017, 8(6):e01843-17.
[44]	TOMICH M, PLANET P J, FIGURSKI D H. The tad locus:Postcards from the widespread colonization island[J]. Nat Rev Microbiol, 2007, 5(5):363-375.

Resistance and Virulence Analysis of Type A: L1 ST128 Pasteurella multocida from Ducks

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 44

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHENG Rui, LIU Zishi, ZHANG Kangyou, YAN Yong, WEI Ling, ZEREN Wengmu, DINGZE Demi, HUANG Jianjun, WANG Li, WEI Yong. Isolation, Identification and Biological Characterization of Colletotrichum jasminigenum in Stems of Peanuts [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2206-2213.
[2]	TIAN Rui, XU Sixiang, XIE Feng, LIU Guangjin, WANG Gang, LI Qingxia, DAI Lei, XIE Guoxin, ZHANG Qiongwen, LU Yajing, WANG Guangwen, WANG Jinxiu, ZHANG Wei. Bioinformatics Analysis of the Genome of Clostridium perfringens Isolated from Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1707-1715.
[3]	CHEN Yuetong, LIU Xiaohan, WANG Zhiyang, ZHAO Yuxin, ZHOU Tiezhong, HU Zengjin, ZHU Yue, WANG Shaohui, TIAN Mingxing, DING Siyu, QI Jingjing, YU Shengqing. Isolation, Identification, Pathogenicity and Drug Susceptibility of Mycoplasma gallisepticum from Dead Chicken Embryos in Large-scale Chicken Farms in Guangdong Province [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 290-299.
[4]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[5]	WANG Jiali, ZHOU Ning, CHEN Xi, YUE Hua, TANG Cheng. Isolation, Identification and Pathogenicity of Canine Adenovirus Type 2 Strain with Natural Deletion in E3 Gene [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2982-2990.
[6]	YUAN Sheng, LI Anqi, Lü Wenke, YANG Lulu, ZHOU Feng, HUANG Liangzong, BAI Aiquan, WEN Feng, HUANG Shujian, GUO Jinyue. Analysis of Variation in Major Virulence-related Genes of a Strain of Pseudorabies Virus and Its Pathogenicity to Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2195-2199.
[7]	LIU Jiankui, XU Ye, LIU Chen, YU Hui, YANG Yuan, HE Le, LI Jiarui, DAN Huijuan, DAI Ailing, YANG Xiaoyan, WEI Chunhua. Genomic Characteristics of Porcine Reproductive and Respiratory Syndrome Virus in Fujian Province from 2017 to 2021 based on Whole Genome [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1579-1589.
[8]	ZHANG Ao, TAN Bin, LIU Kexin, LIU Jiali, ZHANG Shuqin. Genomic Characterization Analysis of a H1N1 Subtype Swine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4866-4871.
[9]	LI Chang, ZHOU Ping, WEN Ping, YANG Keli, LIU Wei, GUO Rui, LIU Zewen, YUAN Fangyan, GAO Ting, SONG Haofei, TIAN Yongxiang, ZHOU Danna. Investigation of the Genome-wide Sequence and Immunogenicity of Capsid Protein in Mice for a Porcine Circovirus 2d Genotype Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4289-4299.
[10]	WANG Meng, ZHANG Senhao, ZHANG Liu, LI Jiaxuan, WANG Xiaona, CUI Wen, JIANG Yanping, ZHOU Han, WANG Li, QIAO Xinyuan, LI Yijing, TANG Lijie. Isolation,Identification and Genetic Evolution Analysis of a Subgroup E Avian Leukemia Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3107-3120.
[11]	XU Lifeng, JIA Chenyu, CHEN Fuzai, FANG Mengyuan, LIU Xiaodan, CHEN Jilong, LI Xunliang. Isolation, Identification and Whole Genome Analysis of a Mycoplasma synoviae Strain from Chicken [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2663-2676.
[12]	WANG Xi, LI Ke, LI Tingcui, YAN Hongya, ZHAO Rong, CHANG Zhishun, LIAO Ming, SUN Minhua, XIN Aiguo. MLST Typing and Drug Resistance Analysis of 75 Salmonella Strains Isolated from Laying Hens [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1626-1631.
[13]	LI Huaming, XIANG Wei, LU Wenbing, LIU Feng, LEI Liancheng, ZHANG Fuxian. Pathogenicity and Drug Sensitivity Analysis of a Porcine Klebsiella pneumoniae Type ST-35 [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4356-4366.
[14]	HE Jian, XING Xiaoyong, WU Xiaochun, WEN Fengqin, ZHANG Yangyang, ZHANG Shengying, LIU Jia, BAO Shijun. Isolation and Identification of a Strain of Pseudomonas aeruginosa from Forest Musk and Analysis of the Whole Genome Sequence [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2569-2578.
[15]	HUANG Chengyu, XU Dequan, FENG Zhe, ZHOU Ling, LIU Min. The Addition of Melatonin Reduces the Oxidative Damage of Pig Spermatogonial Stem Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 1880-1890.