鸭疫里氏杆菌LC1和CX1株的分离鉴定及致病性分析

doi:10.11843/j.issn.0366-6964.2024.09.043

Abstract

Abstract:

In currently study, LC1 and CX1 of Riemerella anatipestifer type 11 were isolated and purified from the brain and liver tissue of 10-day-old ducks, The aim of this study was to isolate, purify, identify and analyze the pathogenicity of the strains from the disease material of Muscovy ducks which suspected to be infected with infectious serositis from a farm in Putian, Fujian Province, China. Brain and liver tissues of 10-day-old diseased Muscovy ducks were collected for strain isolation and purification, and 16S rDNA PCR identification and serotyping were conducted, respectively. The isolates were subjected to 16S rDNA sequencing and genetic evolutionary analyses, and their drug resistance and pathogenicity to ducklings were determined. The 16S rDNA sequence sequencing and genetic evolution of these isolates were analyzed, and their drug resistance and pathogenicity to ducklings were further determined. (Results) The results showed that 2 strains of Riemerella anatipestifer type 11 were obtained and named as LC1 and CX1. The 16S rDNA sequences of LC1 and CX1 showed that the isolates were in the same evolutionary branch as the reference strains of Riemerella anatipestifer Fujian strain RAf115 and Shanghai strain Yb2, with 99.9% similarity. The isolates were sensitive to β-lactams, chloramphenicol, and tetracyclines, but less sensitive to macrolides and aminoglycosides. The pathogenicity assay indicated that the mortality of the two strains (1.3×10⁶ CFU·mL^-1) was 100%, and the lethal ducks showed the typical Riemer's disease with pericarditis, perihepatitis, and enlarged spleen.The pathology results of the HE staining showed that the red-white medullary boundary of the spleen of the lethal ducks disappeared, and a large number of necrotic foci were presented. In addition, myocardial fibers of lethal ducks from the two isolates showed tear-like lesions. These datas demonstrated that two strains of Riemerella anatipestifer type 11 LC1 and CX1 were isolated and characterized, MLST to be ST111 and ST114, respectively, which provide reference basis for the prevention and control of Riemerella anatipestifer disease and subsequent research.

Key words: Riemerella anatipestifer, LC1, CX1, genetic evolution analysis, pathogenicity

CLC Number:

S852.617

Bilin XIE, Zhimin LIN, Binbin LIN, Yijuan XU, Fengqiang LIN, Lu YAN, Huini WU, Cuiting LI, Haiou ZHOU, Zhaolong LI. Isolation, Identification and Pathogenicity Analysis of Riemerella anatipestifer Strain LC1 and CX1[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4196-4203.

Figures/Tables 7

Table 1

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Table 2

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Table 3

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References 20

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名称Name	序列Sequence	长度/bp Length
RAdnaB-F	GAATGATTTCTTCCGAGACAGG	570
RAdnaB-R	AAAGCCTTACTTCGCCCGT
RAgroE-F	TCAAGAGACGCACTTAAAAGAGGTG	519
RAgroE-R	TGTACCTTTAGCCTCTTCCACAGTA
RAgyrA-F	GCCTCATAACTTATCGGAATC	797
RAgyrA-R	AATAGCTTTGTACTCATCACGG
RAmdh-F	ACAGCAGGGTCTCAAGTGG	532
RAmdh-R	TCTGGTCGCAAGCAATAGC
RAgluD-F	TTGTTTCCGCTCGCTCTAAG	769
RAgluD-R	TAATCCCGTTCATCGCCAC
RAgpi-F	GCGATAGATAGTATGTTTTCTGGAG	575
RAgpi-R	TCTACCACCAACCCAGTCC
RArpoB-F	TTAGATCCCATCAAGGCACG	686
RArpoB-R	GAGCAGTTGGCAGGTCAGTT

抗菌素Antibiotic	判定标准/mm Criterion			分离株Isolates
抗菌素Antibiotic	R	I	S	LC1	CX1
青霉素G Penicillin G	≤17	18~20	≥21	24	25
氨苄西林Ampicillin	≤13	14~16	≥17	17	20
阿莫西林Amoxicillin	≤13	14~17	≥18	30	32
头孢曲松Ceftriaxone	≤14	15~17	≥18	31	33
头孢吡肟Cefepime	≤14	15~17	≥18	33	31
庆大霉素Gentamicin	≤12	13~14	≥15	0	0
阿米卡星Amikacin	≤14	15~16	≥17	0	0
新霉素Neomycin	≤12	13~16	≥17	0	0
四环素Tetracycline	≤14	15~18	≥19	30	35
强力霉素Doxycycline	≤12	13~15	≥16	31	32
红霉素Erythromycin	≤13	14~22	≥23	0	0
恩诺沙星Enrofloxacin	≤15	16~20	≥21	0	13
环丙沙星Ciprofloxacin	≤15	16~20	≥21	0	15
复方新诺明Compound sulfamethoxazole	≤9	10~16	≥17	0	0
多黏菌素B Polymyxin B	≤7	8~11	≥12	0	0
氟苯尼考Florfenicol	≤12	13~17	≥18	34	30

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Isolation, Identification and Pathogenicity Analysis of Riemerella anatipestifer Strain LC1 and CX1

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菌株Strain	管家基因House-keeping genes							ST
菌株Strain	dnaB	groEL	gyrA	mdh	gluD	gpi	rpoB	ST
LC1	3	2	4	3	4	30	10	111
CX1	3	2	4	3	4	6	10	114