B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价

doi:10.11843/j.issn.0366-6964.2024.10.031

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (10): 4597-4604.doi: 10.11843/j.issn.0366-6964.2024.10.031

B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价

许壮壮¹^,²(), 王素艳¹(), 陈运通¹, 张涛¹, 于蒙蒙¹, 孟令宅¹, 范文瑞¹, 祁小乐¹, 高玉龙¹^,²^,*()

1. 中国农业科学院哈尔滨兽医研究所动物疫病防控全国重点实验室禽免疫抑制病创新团队, 哈尔滨 150069
2. 黑龙江八一农垦大学动物科技学院, 大庆 163000

收稿日期:2023-12-11 出版日期:2024-10-23 发布日期:2024-11-04
通讯作者: 高玉龙 E-mail:xuzhuangzhuang21@163.com;wangsuyan@caas.cn;gaoyulong@caas.cn
作者简介:许壮壮(1999-), 男, 山东菏泽人, 硕士生, 主要从事禽免疫抑制病研究, E-mail: xuzhuangzhuang21@163.com
王素艳(1990-), 女, 山东济宁人, 助理研究员, 主要从事禽免疫抑制病研究, E-mail: wangsuyan@caas.cn
第一联系人：
许壮壮和王素艳为同等贡献作者
基金资助:
国家肉鸡产业技术体系(CARS-41)

Evaluation of the Immune Efficacy of Attenuated Strain of Subtype B Avian Metapneumovirus Disease on Commercial Broilers

Zhuangzhuang XU¹^,²(), Suyan WANG¹(), Yuntong CHEN¹, Tao ZHANG¹, Mengmeng YU¹, Lingzhai MENG¹, Wenrui FAN¹, Xiaole QI¹, Yulong GAO¹^,²^,*()

1. Avian Immunosuppressive Diseases Division, State Key Laboratory for Animal Disease Control and Prevention Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China
2. College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163000, China

Received:2023-12-11 Online:2024-10-23 Published:2024-11-04
Contact: Yulong GAO E-mail:xuzhuangzhuang21@163.com;wangsuyan@caas.cn;gaoyulong@caas.cn

摘要/Abstract

摘要：

旨在研究B亚型禽偏肺病毒减毒株(LN16-A株)对商品肉鸡的免疫效果。本研究从规模化养殖场选取3周龄的商品肉鸡，将B亚型禽偏肺病毒减毒株(LN16-A株)通过滴鼻方式免疫(200 μL，病毒含量≥10³ TCID₅₀·只^-1)，空白对照组以同样剂量的PBS滴鼻。免疫21 d后，翅下采集血液分离血清，利用ELISA方法及中和试验检测相应抗体；在6周龄时，通过滴鼻方式使用B亚型禽偏肺病毒强毒(LN16-V株)(200 μL，5 000 TCID₅₀·只^-1)进行攻毒，攻毒后连续观察7 d，并利用RT-qPCR检测各组鸡鼻腔拭子的病毒拷贝数，攻毒后第9天各组随机选取2只鸡进行剖杀，采集鼻甲骨、气管和肺组织，通过HE染色制作病理切片，观察组织病理学变化。抗体检测结果显示，在疫苗免疫后21 d，免疫组ELISA抗体和中和抗体阳性率均为100%，平均ELISA抗体效价为1 860，平均中和抗体效价为7.44 log2；临床症状观察结果显示，对照组商品肉鸡出现混浊、黏稠、牵丝状的鼻液等明显临床症状，发病率为100%，而免疫组肉鸡无临床症状；RT-qPCR结果表明，免疫组鸡鼻腔拭子病毒基因组拷贝数相较于对照组降低了78%~96%；病理切片结果显示，对照组肉鸡鼻甲骨、气管以及肺均出现不同程度的病理损伤，免疫组肉鸡各组织器官未见明显病理变化。综上表明，B亚型禽偏肺病毒减毒株(LN16-A株)可以诱导商品肉鸡产生特异性抗体，能显著降低攻毒后的排毒水平，对商品肉鸡具有良好的保护效果。本研究结果将为肉鸡养殖场禽偏肺病毒病的防控提供重要理论指导和技术支撑。

关键词: B亚型禽偏肺病毒, 减毒株, 商品肉鸡, 免疫效果

Abstract:

In order to study the immune effect of attenuated strain of subtype B avian metapneumovirus (LN16-A strain) on commercial broilers, 3-week-old commercial broilers were selected from large-scale farms and immunized with attenuated strain of subtype B avian metapneumovirus (LN16-A strain) by dropping nose (200 μL, virus content≥10³ TCID₅₀/piece), and the blank control group was given the same dose of PBS. Twenty-one days after immunization, blood serum was collected under the wings and corresponding antibodies were detected using ELISA and neutralization tests. At 6-week-old, the B subtype avian metapneumonia virus (LN16-V strain) was administered via nasal drip (200 μL, 5000 TCID₅₀ per chicken) to chickens for virus challenge. After the challenge, the virus copy number of nasal swabs from each group of chickens was continuously observed for 7 days. RT-qPCR was used to detect the virus copy number. On the 9th day after the challenge, 2 chickens from each group were randomly selected for dissection. Nasal turbinate, tracheal, and lung tissues were collected, and pathological sections were made using HE staining to observe the histopathological changes of the tissues. The results of antibody showed that 21 days after immunization, the positive rate of ELISA antibody and neutralizing antibody in the immunization group was 100%, with an average ELISA antibody titer of 1 860 and an average neutralizing antibody titer of 7.44 log2. The results of clinical symptoms showed that the commercial broilers in the control group had obvious clinical symptoms, such as cloudy, viscous, and filamentous nasal juice, and the incidence rate was 100%. While the broilers in the immunization group had no clinical symptoms. In addition, the results of RT-qPCR showed that the virus copy number of nasal swab in the immunization group decreased by 78% to 96%, compared to the control group. The results of pathological sections showed that the turbinate bone, trachea and lung of broilers in the control group had different degrees of pathological damage, while no obvious pathological changes were observed in the organs of broilers in the immunization group. The above results indicate that the attenuated strain of subtype B avian metapneumovirus (LN16-A strain) could induce the production of specific antibodies in commercial broilers, which could significantly reduce the virus shedding level of broilers after challenge, and had a good protective effect on commercial broilers. This study may provide important theoretical guidance and technical support for the prevention and control of avian metapneumovirus disease in large-scale broiler farms.

Key words: subtype B avian metapneumovirus, attenuated strain, commercial broilers, immune effect

中图分类号:

S852.659.5

许壮壮, 王素艳, 陈运通, 张涛, 于蒙蒙, 孟令宅, 范文瑞, 祁小乐, 高玉龙. B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价[J]. 畜牧兽医学报, 2024, 55(10): 4597-4604.

Zhuangzhuang XU, Suyan WANG, Yuntong CHEN, Tao ZHANG, Mengmeng YU, Lingzhai MENG, Wenrui FAN, Xiaole QI, Yulong GAO. Evaluation of the Immune Efficacy of Attenuated Strain of Subtype B Avian Metapneumovirus Disease on Commercial Broilers[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4597-4604.

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参考文献 19

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引物名称 Primer name	序列(5′→3′) Sequences
aMPV/B-F	AATAGTCCTCAAGCAAGTCCTCAGA
aMPV/B-R	CTGTTGTAATTTGACCTGTTCTA CACT
探针Probe	CTGGTGTTATCAGCCTTAGGCTTGACGCT

项目 Item	鸡编号 Number
项目 Item	1	2	3	4	5	6	7	8	9	10
中和抗体效价(log2)	7.76	7.68	7.51	7.50	7.26	7.50	7.41	7.50	7.26	7.00
Neutralizing antibody titer/log2
中和抗体阴(-)或阳性(+)	+	+	+	+	+	+	+	+	+	+
Neutralizing antibody negative (-) or positive (+)
中和抗体阳性率/%	100
Neutralizing antibody positive rate

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B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价

Evaluation of the Immune Efficacy of Attenuated Strain of Subtype B Avian Metapneumovirus Disease on Commercial Broilers

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