产金属β-内酰胺酶猪源ST201型多杀性巴氏杆菌的耐药性和致病性分析

doi:10.11843/j.issn.0366-6964.2024.12.032

Abstract

Abstract:

The study was design to investigate the pathogenic agents of respiratory tract diseases of fattening pigs in a pig breeding enterprise in Jingzhou, Hubei Province. The pathogen was screened and isolated from the diseased pig lung tissue. Morphological observation, physiological and biochemical analysis, 16S rDNA gene sequence, serotype typing, multilocus sequence typing, drug sensitivity test, drug resistance gene sequencing analysis and animal pathogenicity test were performed for the isolated strains. The results showed that the isolated strain Pm0525 was Gram-negative brevibacterium. It was identified as Pasteurella multocida by biochemical assay, PCR and 16S rDNA sequence analysis. The isolated strain was identified as Pasteurella multocida type A. Multilocus sequence typing analysis confirmed that the isolated strain is type ST201 and merged with type ST1615. The isolated strain was sensitive 13 kinds of antibiotics, such as gentamicin, kanamycin and medemycin, and resistant to chloramphenicol, clindamycin, cefuroxime, ceftazidime and tetracycline. The isolated strain showed moderate biofilm formation ability, carrying out four resistance genes, SulⅠ, SulⅡ, tetA and bla-TEM. The isolated strain may play the role of anti-β-lactam antibiotics by producing metal-beta-lactamase (MBL). In addition, traditional Chinese medicine Coptis had obvious inhibitory effect on the isolated strain in vitro. The isolated strain carried 18 virulence genes, and showed strong pathogenicity to mice. This study systematically analyzed the serotype, molecular typing, drug sensitivity and pathogenicity of swine Pasteurella multocida, providing scientific basis and data reference for further research on the transmission, prevention and precise treatment of Pasteurella infection.

Key words: Pasteurella multocida, metallo-beta-lactamase, MLST analysis, drug resistance, pathogenicity

CLC Number:

S852.61

YANG Rongrong, ZHANG Ting, TANG Pingping, HE Shuangfang, ZHAO Miaomiao, LEI Liancheng, ZHANG Fuxian. Analysis of Drug Resistance and Pathogenicity of Metallo-β-Lactamase-producing Porcine ST201 Pasteurella multocida[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5692-5705.

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血清型 Serotype	基因 Gene	引物序列(5′→3′) Primer sequences	片段大小/bp Fragment size
A	hyaD-hyaC	F: TGCCAAAATCGCAGTCAG	1 044
A	hyaD-hyaC	R: TTGCCATCATTGTCAGTG	1 044
B	bcbD	F: CATTTATCCAAGCTCCACC	760
B	bcbD	R: GCCCGAGAGTTTCAATCC	760
D	dcbF	F: TTACAAAAGAAAGACTAGGAGCCC	657
D	dcbF	R: CATCTACCCACTCAACCATATCAG	657
E	ecbJ	F: TCCGCAGAAAATTATTGACTC	511
E	ecbJ	R: GCTTGCTGCTTGATTTTGTC	511
F	fcbD	F: AATCGGAGAACGCAGAAATCAG	851
F	fcbD	R: TTCCGCCGTCAATTACTCTG	851

分离株 Isolate	地区 Region	时间 Year	宿主 Host	来源 Sample	MLST
分离株 Isolate	地区 Region	时间 Year	宿主 Host	来源 Sample	mhadk	mharoA	mhdeoD	mhgdhA	mhg6pd	mhmdh	mhpgi	ST
XJ-PM99	中国	2020	牛	鼻拭子	1	1	1	53	1	1	1	201
XJ-PM100	中国	2020	牛	鼻拭子	1	1	1	53	1	1	1	201
PM0525	中国	2022	猪	鼻拭子	1	1	1	53	1	1	1	201

耐药类型 Types of drug resistance	耐药谱 Antibiotic resistance
耐药	氯霉素Chloramphenicol，克林霉素Clindamycin，头孢呋辛Cefuroxime，头孢他啶Ceftazidime，四环素Tetracycline
中介	红霉素Erythromycin，羧苄西林Carbenicillin，氨苄西林Ampicillin，呋喃唑酮Furazolidone
敏感	庆大霉素Gentamicin，卡那霉素Kanamycin，环丙沙星Ciprofloxacin，麦迪霉素Midecamycin，新霉素Neomycin，氧氟沙星Ofloxacin，诺氟沙星Norfloxacin，哌拉西林Piperacillin，丁胺卡那Amikacin，头孢氨苄Cephalexin，头孢咪唑Cefpimizole，头孢拉定Cefradine，头孢哌酮Cefoperazone

中药 Traditional Chinese medicine	抑菌圈直径/mm IZD/mm	敏感性 Sensitivity	中药 Traditional Chinese medicine	抑菌圈直径/mm IZD/mm	敏感性 Sensitivity
连翘 Forsythia suspensa	7	+	乌梅 Dark plum	8	+
黄连 Coptis chinensis	19	++	秦皮 Ash bark	7	+
五倍子 Gallnut	12	++	荆芥 Schizonepeta	10	++
肉桂 Cinnamomum cassia	7	+	五味子 Schisandra chinensis	0	-
款冬花 Coltsfoot flower	2	-	蒲公英 Dandelion	2	-
金银花 Honeysuckle	0	-	枳壳 Fructus Aurantii	0	-
炒僵蚕 Fried Silkworm	0	-	板蓝根 Radix isatidis	0	-
金蝎 Golden Scorpion	3	-	羌活 Notopterygium root	0	-
桑皮 Mulberry bark	0	-	炙白附子 Roasted White Aconite	4	-
薄荷 Mint	0	-	鱼腥草 Houttuynia cordata	0	-
苏子 Perillaseed	0	-	贝川母 Beichuan mother	0	-
桑白皮 Morus alba	0	-	胖大海 scaphium scaphigerum	0	-
黄芩 Scutellaria baicalensis	0	-	首乌 Polygonum multiflorum	1	-
茯苓 Poria cocos	0	-	罗汉果 Siraitia grosvenorii	0	-

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Analysis of Drug Resistance and Pathogenicity of Metallo-β-Lactamase-producing Porcine ST201 Pasteurella multocida

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