2022年中国南方部分地区猪肺炎支原体感染状况的调查与分析

doi:10.11843/j.issn.0366-6964.2024.07.025

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (7): 3064-3074.doi: 10.11843/j.issn.0366-6964.2024.07.025

2022年中国南方部分地区猪肺炎支原体感染状况的调查与分析

高一鸣¹^,²(), 陈国盛¹^,², 倪诗婷¹^,², 佟泽¹^,², 王豪男³, 杨钒¹^,², 杨丽君¹^,², 莫玉鹏³^,*(), 谭臣¹^,²^,*()

1. 华中农业大学动物医学院，武汉 430070
2. 生殖健康养殖协同创新中心，武汉 430070
3. 桂林力源粮油食品集团有限公司，桂林 541012

收稿日期:2023-08-28 出版日期:2024-07-23 发布日期:2024-07-24
通讯作者: 莫玉鹏,谭臣 E-mail:gym1994@webmail.hzau.edu;741623673@qq.com;tanchen@mail.hzau.edu.cn
作者简介:高一鸣(1995-)，男，山西太原人，硕士，主要从事动物传染病研究，E-mail: gym1994@webmail.hzau.edu
基金资助:
国家重点研发计划(2022YFD1800900);国家生猪产业技术体系岗位科学家项目(CARS-35)

Investigation and Analysis of Infection Status of Mycoplasma hyopneumoniae in Pigs in Selected Areas of Southern China, 2022

Yiming GAO¹^,²(), Guosheng CHEN¹^,², Shiting NI¹^,², Ze TONG¹^,², Haonan WANG³, Fan YANG¹^,², Lijun YANG¹^,², Yupeng MO³^,*(), Chen TAN¹^,²^,*()

1. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
2. The Collaborative Innovation Centre for Sustainable Pig Production, Wuhan 430070, China
3. Guilin Li Yuan Grain and Oil Food Group Co., Ltd, Guilin 541012, China

Received:2023-08-28 Online:2024-07-23 Published:2024-07-24
Contact: Yupeng MO, Chen TAN E-mail:gym1994@webmail.hzau.edu;741623673@qq.com;tanchen@mail.hzau.edu.cn

摘要/Abstract

摘要：

猪肺炎支原体(Mycoplasma hyopneumoniae，Mhp)是猪地方性肺炎(EP)的原发性病原，可导致猪咳嗽、气喘、消瘦，并因猪的饲料转化率升高、日增重减少、出栏时间延长，从而使养殖成本大大增加，给养猪业造成重大经济损失。本研究旨在对中国南方部分地区猪肺炎支原体的感染状况进行调查与分析。从中国南方部分地区21个规模化种猪场定点采样，首先通过病原检测比较鼻拭子和喉拭子的采样效果，随后以简单随机抽样结合分层抽样确定各猪场样本量。共收集母猪喉拭子样品1 478份，并将调查的猪群按胎次分为“0~1胎母猪”和“≥2胎母猪”，通过实时荧光定量PCR进行病原检测，通过多位点序列分型(MLST)结合保守的p36基因及非保守的p97和p146基因进行分子流行病学分析。检测结果表明，鼻拭子样品的检出率为16.67%，喉拭子样品的检出率为56.67%，且Ct值更低。猪肺炎支原体感染的场检出率为66.67%，但各场感染压力较小，检出率为1.39%~18.57%。其中“0~1胎母猪”检出率为10.05%；“≥2胎母猪”检出率为2.17%。分子流行病学结果显示，7个猪场的流行菌株均为同一序列类型(ST128)，不同猪场间的流行菌株高度同源。对于猪肺炎支原体的病原学检测，喉拭子是比鼻拭子更灵敏且有效的活体样品类型，母猪胎次和生猪调运引种是规模化种猪场感染猪肺炎支原体的重要因素。本研究为猪肺炎支原体的防控和净化工作提供理论基础。

关键词: 猪肺炎支原体, 定点采样, 感染状况, 分子流行病学

Abstract:

Mycoplasma hyopneumoniae (Mhp) is the primary pathogen of endemic pneumonia (EP) in pigs, which causes coughing, shortness of breath, and wasting, resulting in higher feed conversion ratio, lower daily weight gain, longer farrowing time, and much higher farming costs, which cause significant economic losses to the pig farming industry. The aim of this study was to investigate and analyze the infection status of Mycoplasma hyopneumoniae in selected areas of southern China. A total of 21 large-scale pig farms in southern China were fixed-point sampled, and the sampling efficiency of nasal swabs and laryngeal swabs was compared through pathogen detection. Subsequently, the sample size for each farm was determined using simple random sampling combined with stratified sampling. A total of 1 478 laryngeal swab samples from sows were collected, and the surveyed pig populations were divided into "0-1 parity sows" and "≥2 parity sows". Pathogen detection was performed using real-time fluorescence quantitative PCR, and molecular epidemiological analysis was conducted using multi-locus sequence typing (MLST) combined with conserved p36 gene and non-conserved p97 and p146 genes. The results showed that the detection rate of nasal swab samples was 16.67%, and the detection rate of laryngeal swab samples was 56.67%, with lower Ct values. The detection rate of Mycoplasma hyopneumoniae infection in the field was 66.67%, but the infection pressure in each field was relatively low, with detection rates ranging from 1.39% to 18.57%. Among them, the detection rate of "0-1 parity sows" was 10.05%; the detection rate of "≥2 parity sows" was 2.17%. Molecular epidemiological results showed that the prevalent strains in 7 farms were of the same sequence type (ST128), and the genetic variation of the prevalent strains among the different farms was small and highly homogeneous. For the pathogen detection of Mycoplasma hyopneumoniae, laryngeal swabs are a more sensitive and effective sample type in vivo than nasal swabs. Parity of sows and the transportation of pigs are important factors for the infection of Mycoplasma hyopneumoniae in large-scale breeding farms. This study provides a theoretical basis for the prevention and control of Mycoplasma hyopneumoniae.

Key words: Mycoplasma hyopneumoniae, fixed-point sampling, infection status, molecular epidemiology

中图分类号:

S852.62

高一鸣, 陈国盛, 倪诗婷, 佟泽, 王豪男, 杨钒, 杨丽君, 莫玉鹏, 谭臣. 2022年中国南方部分地区猪肺炎支原体感染状况的调查与分析[J]. 畜牧兽医学报, 2024, 55(7): 3064-3074.

Yiming GAO, Guosheng CHEN, Shiting NI, Ze TONG, Haonan WANG, Fan YANG, Lijun YANG, Yupeng MO, Chen TAN. Investigation and Analysis of Infection Status of Mycoplasma hyopneumoniae in Pigs in Selected Areas of Southern China, 2022[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3064-3074.

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类型 Type	引物名称 Primer name	引物序列(5′-3′) Primer sequence	退火温度/℃ Tm	产物大小/bp Product size
MLST管家基因扩增引物 MLST housekeeping gene amplification primer	adk-F	CCCAGGAGAACAGACTGAAGA	59	1 148
	adk-R	GCTGGTTTTGCATTTCGGCT
	rpoB-F	GCGGTTCACTTCGGGTAGAA	59	1 848
	rpoB-R	TAGCAACAAGTGGGGCTTCTG
	tpiA-F	CCTTCTAATTTCGTCATCGCGG	59	1 225
	tpiA-R	AGTGCAATCTGAACGCTTGT
MLST管家基因测序引物 MLST housekeeping gene sequencing primer	adk-SF	AGCAACAACTACACTTGAAACGA	59	874
	adk-SR	TGCCAGCAATTTTCAACTGG
	rpoB-SF	AGTGATGATCCAGATTCGCTTGT	59	640
	rpoB-SR	TGTAATTGACTTGTTCGGCATC
	tpiA-SF	CCTTCTAATTTCGTCATCGCGG	59	1 225
	tpiA-SR	AGTGCAATCTGAACGCTTGT
p146基因扩增/ 测序引物 p146 gene amplification/ sequencing primer	P146S-F	CGAACGTGGATGGATGTCAACT	59	870~910
p146基因扩增/ 测序引物 p146 gene amplification/ sequencing primer	P146S-R	GTCTGACCCATTTGTTGGCCATTAT
p36基因扩增/ 测序引物 p36 gene amplification/ sequencing primer	P36FL-F	AACACGCCACCAGGAATTGA	59	1 775
p36基因扩增/ 测序引物 p36 gene amplification/ sequencing primer	P36FL-R	AGGCGTGCAACCTATAGCAG
p97基因扩增/ 测序引物 p97 gene amplification/ sequencing primer	P97R1-F	GCCTATGAATTCGGGGCCTTTA	59	1 189~1 417
p97基因扩增/ 测序引物 p97 gene amplification/ sequencing primer	P97R1-R	CGGTCATCATTGGGTGGCTA

编号 No.	猪场A Farm A				猪场B Farm B
	鼻拭子 Nasal swab		喉拭子 Laryngeal swab		鼻拭子 Nasal swab		喉拭子 Laryngeal swab
	Ct值 Ct value	结果判定 Result	Ct值 Ct value	结果判定 Result	Ct值 Ct value	结果判定 Result	Ct值 Ct value	结果判定 Result
1	/	-	34.36	+	/	-	/	-
2	36.03	+	28.79	+	/	-	/	-
3	/	-	29.50	+	/	-	/	-
4	36.97	+	28.95	+	/	-	/	-
5	36.98	+	27.45	+	/	-	/	-
6	36.55	+	28.72	+	/	-	/	-
7	/	-	26.17	+	/	-	/	-
8	/	-	28.41	+	/	-	/	-
9	/	-	23.20	+	/	-	/	-
10	32.48	+	27.51	+	/	-	/	-
11	/	-	32.84	+	/	-	/	-
12	/	-	31.96	+	/	-	/	-
13	/	-	27.86	+	/	-	/	-
14	/	-	20.82	+	/	-	33.49	+
15	/	/	/	/	/	-	35.75	+
16	/	/	/	/	/	-	33.37	+
检出率/% Detection rate	35.71(5/14)		100.00(14/14)		0.00(0/16)		18.75(3/16)

猪场 Farm	位置 Location	0~1胎母猪检出率 Detection rate of 0-1 parity sows	≥2胎母猪检出率 Detection rate of ≥2 parity sows	母猪检出率^* Detection rate of all sows	场检出率 Field detection rate
A	广西	20.83(5/24)	0.00(0/46)	7.14(5/70)	66.67
B	广西	16.67(4/24)	2.17(1/46)	7.14(5/70)	(14/21)
C	广西	33.33(4/12)	15.52(9/58)	18.57(13/70)
D	广西	0.00(0/31)	0.00(0/39)	0.00(0/70)
E	广东	0.00(0/11)	0.00(0/59)	0.00(0/70)
F	广西	18.75(3/16)	1.85(1/54)	5.71(4/70)
G	广西	1.92(1/52)	0.00(0/18)	1.43(1/70)
H	湖南	0.00(0/18)	0.00(0/52)	0.00(0/70)
I	广西	20.59(7/34)	2.78(1/36)	11.43(8/70)
J	广西	1.39(1/72)	/	1.39(1/72)
K	贵州	0.00(0/49)	0.00(0/21)	0.00(0/70)
L	广西	12.50(4/32)	0.00(0/38)	5.71(4/70)
M	广西	47.37(9/19)	3.92(2/51)	15.71(11/70)
N	贵州	7.89(3/38)	7.89(3/38)	7.89(6/76)
O	广西	29.03(9/31)	0.00(0/39)	12.86(9/70)
P	海南	18.57(13/70)	/	18.57(13/70)
Q	广西	10.53(2/19)	1.96(1/51)	4.29(3/70)
R	广西	0.00(0/23)	0.00(0/47)	0.00(0/70)
S	海南	0.00(0/7)	0.00(0/63)	0.00(0/70)
T	四川	11.63(5/43)	11.11(3/27)	11.43(8/70)
U	云南	0.00(0/22)	0.00(0/48)	0.00(0/70)
总计Total		10.05 (65/647)	2.17 (18/831)	5.62 (83/1 478)

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2022年中国南方部分地区猪肺炎支原体感染状况的调查与分析

Investigation and Analysis of Infection Status of Mycoplasma hyopneumoniae in Pigs in Selected Areas of Southern China, 2022

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