宁夏部分地区犊牛腹泻样本大肠杆菌分离菌株的毒力因子检测与分析

doi:10.11843/j.issn.0366-6964.2024.07.044

摘要/Abstract

摘要：

旨在调查宁夏地区犊牛腹泻性大肠杆菌(Escherichia coli，E. coli)流行情况及毒力因子流行特点。对宁夏地区共采集332份腹泻犊牛直肠粪便进行病原分离，利用PCR方法对156株大肠杆菌分离株进行13种相关毒力基因检测。结果共分离出156株大肠杆菌(46.99%)，共检出136株含有毒力因子的大肠杆菌，表明目前宁夏地区腹泻犊牛大肠杆菌阳性率较高，毒力因子组合复杂，优势毒力因子为irp2。

关键词: 犊牛, 腹泻性, 大肠杆菌, 毒力因子

Abstract:

The aim of this study was to investigate the prevalence and virulence factors of Escherichia coli (E. coli) in calves in Ningxia region. A total of 332 samples of diarrhea calf rectal feces were collected in Ningxia region for pathogen isolation, and 156 strains of E. coli were detected for 13 related virulence genes using PCR method. The results show that a total of 156 strains (46.99%) of E. coli were isolated, and a total of 136 strains containing virulence factors were detected. This indicates that the current positive rate of E. coli in diarrhea calves in Ningxia is relatively high, and the combination of virulence factors is complex, with irp2 being the dominant virulence factor.

Key words: calves, diarrhea, Escherichia coli, toxicity factor

中图分类号:

S852.612

王玮, 王景松, 郭亚男, 高乐, 王玲玲, 陈灿, 王健霖, 王建东, 马科, 李继东. 宁夏部分地区犊牛腹泻样本大肠杆菌分离菌株的毒力因子检测与分析[J]. 畜牧兽医学报, 2024, 55(7): 3261-3266.

Wei WANG, Jingsong WANG, Ya'nan GUO, Le GAO, Lingling WANG, Can CHEN, Jianlin WANG, Jiandong WANG, Ke MA, Jidong LI. Toxicity Factor Detection and Analysis of Escherichia coli Isolated Strains from Calf Diarrhea Samples in Some Areas of Ningxia[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3261-3266.

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引物名称 Primers	引物序列(5′→3′) Primer sequence	扩增片段 Amplified fragment and size	片段大小/bp Fragment size
K88/F41-F	GATTGCTACGTTCAGCGGAGCG	K88	841
K88-R	GATGAAAGACTCTGATTGCA
F41-R	TCTGAGGTCATCCCAATTGTGG	F41	628
K99-F	CTGAAAAAAACACTGCTAGCTATT	K99	543
K99-R	CATATAAGTGACTAAGAAGGATGC
987P-F	GTTACTGCCAGTCTATGCCAAGTG	987P	463
987P-R	TCGGTGTACCTGCTGAACGAATAG
fedA-F	CATGATGGATCCATGAAAAGACTAGTGTTTATTTCTT	F18 fedA	513
fedA-R	CATGATGAATTCTTACTTGTAAGTAACCGCGTAAGCC
HPI(irp2)-F	AAGGATTCGCTGTTACCGGAC	irp2	280
HPI(irp2)-R	TCGTCGGGCAGCGTTTCTTCT
LEE(eae)-F	ATATCCGTTTTAATGGCTATCT	eae	425
LEE(eae)-R	AATCTTCTGCGTACTGTGTTCA
ST1-F	GGGTTGGCAATTTTTATTTCTGTA	ST1	183
ST1-R	ATTACAACAAAGTTCACAGCAGTA
ST2-F	ATGTAAATACCTACAACGGGTGAT	ST2	360
ST2-R	TATTTGGGCGCCAAAGCATGCTCC
LT1-F	TAGAGACCGGTATTACAGAAATCTGA	LT1	282
LT1-R	TCATCCCGAATTCTGTTATATATGTC
VT1-F	ATTCGCTGAATGTCATTCGCT	stx1	664
VT1-R	ACGCTTCCCAGAATTGCATTA
VT2all-F	ATTCGCTGAATGTCATTCGCT	stx1/stx2e	484
VT2all-R	ACGCTTCCCAGAATTGCATTA
SLT2-F	GAATGAAGAAGATGTTTATAGCGG	stx2	281
SLT2-R	GGTTATGCCTCAGTCATTATTAA

地区 Area	合计 Total		规模场 Scale field			散养户 Casual care households
地区 Area	阳性数 Positive number	阳性率/% Positive rate	样品数 Number of samples	阳性数 Positive number	阳性率/% Positive rate	样品数 Number of samples	阳性数 Positive number	阳性率/% Positive rate
西吉县 Xiji County	22	47.83	40	18	45.00	6	4	66.67
青铜峡 Qingtongxia	58	46.03	98	46	46.94	28	12	42.86
利通区 Litong District	8	30.77	22	6	27.27	4	2	50.00
红寺堡 Hongsi Fort	38	52.78	60	28	46.67	12	10	83.33
同心 Tongxin	30	48.39	56	26	46.43	6	4	66.67
总计 Total	156	46.99	276	124	44.93	56	32	57.14

毒力基因 Virulence gene	K88	K99	HPI (irp2)	LEE (eae)	ST1	ST2	VT1	VT2all	SLT2
检出数 Detection number	30	6	82	40	6	2	38	34	20
检出率/% Detection rate	19.23	3.85	52.56	25.64	3.85	1.28	24.36	21.79	12.82

项目 Project	毒力基因 Virulence gene	菌株数/个 Number of strains	检出率/% Detection rate
未检出毒力基因 No virulence genes detected		20	12.82
单一毒力基因 Single virulence gene	HPI(irp2)	56	35.90
	LEE(eae)	8	5.13
	VT1	8	5.13
	VT2all	6	3.85
	K88	2	1.28
两个毒力基因 Two virulence genes	K88、HPI(irp2)	6	3.85
	HPI(irp2)、VT2all	2	1.28
	K88、LEE(eae)	2	1.28
	K88、VT2all	2	1.28
	VT1、SLT2	2	1.28
	VT1、VT2all	2	1.28
	HPI(irp2)、LEE(eae)	2	1.28
	HPI(irp2)、VT1	2	1.28
三个毒力基因 Three virulence genes	K88、HPI(irp2)、LEE(eae)	6	3.85
	VT1、VT2all、SLT2	4	2.56
	LEE(eae)、VT1、VT2all	4	2.56
	LEE(eae)、VT1、SLT2	4	2.56
	K88、K99、LEE(eae)	2	1.28
	K99、HPI(irp2)、VT2all	2	1.28
三个以上毒力基因 More than three virulence genes	ST1、VT1、VT2all、SLT2	2	1.28
	K88、LEE(eae)、VT1、VT2all	2	1.28
	K88、HPI(irp2)、LEE(eae)、ST1	2	1.28
	K88、LEE(eae)、VT1、VT2all、SLT2	2	1.28
	K99、LEE(eae)、VT1、VT2all、SLT2	2	1.28
	K88、HPI(irp2)、LEE(eae)、ST2、VT1、VT2all、SLT2	2	1.28
	K88、HPI(irp2)、LEE(eae)、ST1、VT1、VT2all、SLT2	2	1.28

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