B亚型禽偏肺病毒病弱毒疫苗对商品蛋鸡的免疫效果

doi:10.11843/j.issn.0366-6964.2025.08.033

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (8): 3958-3966.doi: 10.11843/j.issn.0366-6964.2025.08.033

B亚型禽偏肺病毒病弱毒疫苗对商品蛋鸡的免疫效果

薛晓晓¹^,²(), 孟令宅², 王素艳², 于蒙蒙², 陈运通², 祁小乐², 李留安¹, 于晓雪¹^,*(), 高玉龙²^,*()

1. 天津农学院动物科学与动物医学学院天津市农业动物繁育与健康养殖重点实验室, 天津 300392
2. 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

收稿日期:2024-10-30 出版日期:2025-08-23 发布日期:2025-08-28
通讯作者: 于晓雪,高玉龙 E-mail:1213273476@qq.com;yuxiaoxue1990@163.com;gaoyulong@caas.cn
作者简介:薛晓晓(1993-)，女，河北邯郸人，硕士生，主要从事禽免疫抑制病研究，E-mail：1213273476@qq.com
基金资助:
国家重点研发计划资助项目(2023YFE0106100)；国家肉鸡产业技术体系项目(CARS-41)

Study on Immune Effect of Subtype B Attenuated Avian Metapneumovirus Disease Vaccine on Commercial Layer Chickens

XUE Xiaoxiao¹^,²(), MENG Lingzhai², WANG Suyan², YU Mengmeng², CHEN Yuntong², QI Xiaole², LI Liuan¹, YU Xiaoxue¹^,*(), GAO Yulong²^,*()

1. Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China
2. Avian Immunosuppressive Disease Division, State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China

Received:2024-10-30 Online:2025-08-23 Published:2025-08-28
Contact: YU Xiaoxue, GAO Yulong E-mail:1213273476@qq.com;yuxiaoxue1990@163.com;gaoyulong@caas.cn

摘要/Abstract

摘要：

为评价B亚型禽偏肺病毒病弱毒疫苗对商品蛋鸡免疫保护效果，本研究用实验室前期制备的B亚型禽偏肺病毒病弱毒疫苗(LN16-A株)，通过滴鼻方式以400 TCID₅₀ ·只^-1的剂量接种21日龄的商品蛋鸡。免疫后7、14和21 d分别采血分离血清，测定ELISA抗体和中和抗体效价，结果显示，免疫后21 d免疫组血清抗体阳性率为90%，中和抗体平均效价为7.51 log2。免疫后21 d，使用B亚型禽偏肺病毒强毒(LN16-V株)进行攻毒保护评价，攻毒剂量为5 000 TCID₅₀ ·只^-1，结果显示，攻毒对照组鸡出现混浊或黏稠样鼻涕，发病率为80%，免疫组鸡均未发病；采用RT-qPCR方法测定各组鸡1~7 d腭裂拭子中的病毒拷贝数，结果显示，免疫组病毒载量仅为攻毒对照组的1/100；组织病理学结果显示，攻毒对照组鸡的鼻甲骨及气管等组织出现不同程度的病理损伤，而免疫组鸡均未出现任何病理变化。以上结果表明, B亚型禽偏肺病毒病弱毒疫苗(LN16-A株)对商品蛋鸡具有良好的免疫保护效果。本研究为控制蛋鸡群B亚型禽偏肺病毒的流行和制定预防B亚型禽偏肺病的免疫程序提供了理论依据。

关键词: B亚型禽偏肺病毒, 弱毒疫苗, 商品蛋鸡, 免疫效果

Abstract:

To evaluate the protective efficacy of the attenuated vaccine against subtype B avian metapneumovirus on commercial layer chickens, the attenuated vaccine against subtype B avian metapneumovirus (LN16-A strain) prepared in the laboratory was used to immunize 21-day-old commercial layer chickens by intranasal administration at a dose of 400 TCID₅₀/chicken. Samples were collected on 7, 14, and 21 days post-immunization to separate the serum and measure the ELISA antibody and neutralizing antibody titers. At 21 days after immunization, the results showed the positive rate of ELISA antibody was 90% and the average neutralizing antibody titer was 7.51 log2 in the immunization group. At 21 days post-immunization, the commercial layer chickens were attacked by subtype B avian metapneumovirus (LN16-V strain) at a dose of 5 000 TCID₅₀ per chicken. The results indicated that chickens in the challenge control group had cloudy or viscous nose with an 80% incidence rate, while chickens in the immune group showed no clinical symptoms. RT-qPCR was used to measure virus copies in palatine swabs collected from chickens in each group from 1 to 7 days. The results showed that the viral loads in the immune group decreased by 175 folds compared with that in the challenge control group. And the histopathological examination results showed that varying degrees of pathological damage in tissues such as the turbinate bone and trachea of chickens were found in the challenge control group, while no obvious pathological changes of chickens were found in the immune group. The above results indicate that the attenuated vaccine against subtype B avian metapneumovirus (LN16-A strain) can provide protection for commercial layer chickens. And this study further provides a theoretical basis for controlling the prevalence of subtype B avian metapneumovirus in layer flocks and formulating an immunization program to prevent subtype B avian metapneumovirus disease.

Key words: subtype B avian metapneumovirus, attenuated vaccine, commercial layer chickens, immune effect

中图分类号:

S852.659.5

薛晓晓, 孟令宅, 王素艳, 于蒙蒙, 陈运通, 祁小乐, 李留安, 于晓雪, 高玉龙. B亚型禽偏肺病毒病弱毒疫苗对商品蛋鸡的免疫效果[J]. 畜牧兽医学报, 2025, 56(8): 3958-3966.

XUE Xiaoxiao, MENG Lingzhai, WANG Suyan, YU Mengmeng, CHEN Yuntong, QI Xiaole, LI Liuan, YU Xiaoxue, GAO Yulong. Study on Immune Effect of Subtype B Attenuated Avian Metapneumovirus Disease Vaccine on Commercial Layer Chickens[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3958-3966.

图/表 8

表 1

图 1

图 2

图 3

图 4

图 5

图 6

图 7

参考文献 28

1	TOQUIN D , BAYON-AUBOYER M H , SENNE D A , et al. Lack of antigenic relationship between French and recent North American non-A/non-B turkey rhinotracheitis viruses[J]. Avian Dis, 2000, 44 (4): 977- 982. doi: 10.2307/1593075
2	BAYON-AUBOYER M H , ARNAULD C , TOQUIN D , et al. Nucleotide sequences of the F, L and G protein genes of two non-A/non-B avian pneumoviruses (APV) reveal a novel APV subgroup[J]. J Gen Virol, 2000, 81 (Pt 11): 2723- 2733.
3	CHACON J L , BRANDAO P E , BUIM M , et al. Detection by reverse transcriptase-polymerase chain reaction and molecular characterization of subtype B avian metapneumovirus isolated in Brazil[J]. Avian Pathol, 2007, 36 (5): 383- 387. doi: 10.1080/03079450701589142
4	GOYAL S M , CHIANG S J , DAR A M , et al. Isolation of avian pneumovirus from an outbreak of respiratory illness in Minnesota turkeys[J]. J Vet Diagn Invest, 2000, 12 (2): 166- 168. doi: 10.1177/104063870001200214
5	LEE E H , SONG M S , SUIN J Y , et al. Genetic characterization of avian metapneumovirus subtype C isolated from pheasants in a live bird market[J]. Virus Res, 2007, 128 (1-2): 18- 25. doi: 10.1016/j.virusres.2007.03.029
6	BANET-NOACH C , SIMANOV L , PERK S . Characterization of Israeli avian metapneumovirus strains in turkeys and chickens[J]. Avian Pathol, 2005, 34 (3): 220- 226. doi: 10.1080/03079450500112625
7	朱艳梅, 宫晓, 郭伟伟, 等. 2012年-2015年我国部分地区禽偏肺病毒的分子流行病学分析[J]. 动物医学进展, 2016, 37 (10): 30- 34.
	ZHU Y M , GONG X , GUO W W , et al. Molecular epidemiology analysis of aMPV in some regions in China during 2012 to 2015[J]. Progress in Veterinary Medicine, 2016, 37 (10): 30- 34.
8	肖志宇, 王素春, 赵成龙, 等. 2020年我国部分地区禽偏肺病毒流行病学调查[J]. 中国预防兽医学报, 2022, 44 (7): 712- 717.
	XIAO Z Y , WANG S C , ZHAO C L , et al. Epidemiological investigation of avian metapneumovirus in some areas of China in 2020[J]. Chinese Journal of Preventive Veterinary Medicine, 2022, 44 (7): 712- 717.
9	张丹俊, 戴银, 赵瑞宏, 等. 安徽省部分地区鸡群禽偏肺病毒感染的血清学调查[J]. 动物医学进展, 2017, 38 (2): 126- 129.
	ZHANG D J , DAI Y , ZHAO R H , et al. Serological survey on avian metapneumovirus infection in chicken flocks in Anhui province[J]. Progress in Veterinary Medicine, 2017, 38 (2): 126- 129.
10	COOK J K , CHESHER J , ORTHEL F , et al. Avian pneumovirus infection of laying hens: experimental studies[J]. Avian Pathol, 2000, 29 (6): 545- 556. doi: 10.1080/03079450020016788
11	YUHUAN W , FU Q . Analysis on production efficiency of laying hens in China-based on the survey data of five provinces[J]. J Agricult Sci, 2019, 11 (9): 280- 286.
12	包媛玲, 何锡栋, 于蒙蒙, 等. B亚型禽偏肺病毒对蛋鸡的致病性研究[J]. 中国家禽, 2021, 43 (6): 25- 30.
	BAO Y L , HE X D , YU M M , et al. Pathogenicity of avian metapneumovirus subtype B on layer chicken[J]. China Poultry, 2021, 43 (6): 25- 30.
13	王友, 宋新宇, 王海旺, 等. 禽偏肺病毒灭活疫苗攻毒保护效力评估实验[J]. 中国动物保健, 2023, 25 (4): 115- 116.
	WANG Y , SONG X Y , WANG H W , et al. Evaluation of the protective efficacy of inactivated avian metapneumovirus vaccine against challenge[J]. Animal Health in China, 2023, 25 (4): 115- 116.
14	包媛玲, 宋守生, 于蒙蒙, 等. B亚型禽偏肺病毒病灭活疫苗对商品蛋鸡的免疫效果研究[J]. 中国预防兽医学报, 2022, 44 (5): 521- 525.
	BAO Y L , SONG S S , YU M M , et al. Study on immune effect of inactivated vaccine of avian metapneumovirus (Subtype B) disease on commercial layer chickens[J]. Chinese Journal of Preventive Veterinary Medicine, 2022, 44 (5): 521- 525.
15	许壮壮, 王素艳, 陈运通, 等. 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. doi: 10.11843/j.issn.0366-6964.2024.10.031
	XU Z Z , WANG S Y , CHEN Y T , et al. 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. doi: 10.11843/j.issn.0366-6964.2024.10.031
16	MENG L Z , YU M M , WANG S Y , et al. A novel live attenuated vaccine candidate protects chickens against subtype B avian metapneumovirus[J]. J Integr Agr, 2024, 23 (5): 1658- 1670. doi: 10.1016/j.jia.2023.10.025
17	YU M , XING L , CHANG F , et al. Genomic sequence and pathogenicity of the first avian metapneumovirus subtype B isolated from chicken in China[J]. Vet Microbiol, 2019, 228, 32- 38. doi: 10.1016/j.vetmic.2018.11.009
18	包媛玲. B亚型禽偏肺病毒病灭活疫苗制备及免疫效力评价[D]. 哈尔滨: 中国农业科学院, 2021.
	BAO Y L. Preparation and immune efficacy evaluation of inactivated vaccine against avian metapneumovirus disease subtype B[D]. Harbin: Chinese Academy of Agricultural Sciences, 2021. (in Chinese)
19	CECCHINATO M , LUPINI C , POGORELTSEVA O S M , et al. Development of a real-time RT-PCR assay for the simultaneous identification, quantitation and differentiation of avian metapneumovirus subtypes A and B[J]. Avian Pathol, 2013, 42 (3): 283- 289. doi: 10.1080/03079457.2013.788130
20	徐仕忠, 秦云, 于鹏, 等. 中国鸡群禽偏肺病毒感染的流行病学调查报告[J]. 中国家禽, 2013, 35 (6): 59- 60.
	XU S Z , QIN Y , YU P , et al. Epidemiological investigation of avian metapneumovirus infection in chickens in China[J]. China Poultry, 2013, 35 (6): 59- 60.
21	陈秀琴, 郑敏, 黄梅清, 等. 福建省规模化鸡场禽偏肺病毒感染情况的血清学调查[J]. 动物医学进展, 2021, 42 (1): 130- 133.
	CHEN X Q , ZHENG M , HUANG M Q , et al. Serosurvey of avian metapneumovirus in scaled chicken farms of Fujian[J]. Chinese Journal of Preventive Veterinary Medicine, 2021, 42 (1): 130- 133.
22	张丽华, 李和鸣, 袁园, 等. 广西地区规模化鸡场禽偏肺病毒感染情况的血清学调查[J]. 中国预防兽医学报, 2017, 39 (6): 439- 442.
	ZHANG L H , LI H M , YUAN Y , et al. Survey of avian metapneumovirus in chickens of Guangxi region[J]. Chinese Journal of Preventive Veterinary Medicine, 2017, 39 (6): 439- 442.
23	HASSAN M S H , ABDUL-CAREEM M F . Avian viruses that impact table egg production[J]. Animals (Basel), 2020, 10 (10): 1747.
24	贠炳岭, 刘在斯, 吴关, 等. 我国部分地区种鸡禽肺病毒感染的血清学调查[J]. 中国家禽, 2012, 34 (12): 64- 65.
	YUN B L , LIU Z S , WU G , et al. Serological survey of pulmonary virus infection in breeder chickens in some areas of China[J]. China Poultry, 2012, 34 (12): 64- 65.
25	于蒙蒙, 包媛玲, 王素艳, 等. B亚型禽偏肺病毒的分离鉴定及致病性研究[J]. 畜牧兽医学报, 2022, 53 (10): 3540- 3549. doi: 10.11843/j.issn.0366-6964.2022.10.026
	YU M M , BAO Y L , WANG S Y , et al. Isolation, identification and pathogenicity of subgroup B avian metapneumovirus[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53 (10): 3540- 3549. doi: 10.11843/j.issn.0366-6964.2022.10.026
26	PAUDEL S , EASWARAN M , JANG H , et al. Immunization with avian metapneumovirus harboring chicken Fc induces higher immune responses[J]. Virus Res, 2016, 220, 129- 135.
27	YOUN H N , NOH J Y , KIM M S , et al. Efficacy of a novel avian metapneumovirus live vaccine candidate based on vaccination route and age[J]. Poult Sci, 2021, 100 (3): 100528.
28	KHALID Z , HE L , YU Q , et al. Enhanced protection by recombinant Newcastle disease virus expressing infectious bronchitis virus spike ectodomain and chicken granulocyte-macrophage colony-stimulating factor[J]. Avian Dis, 2021, 65 (3): 364- 372.

引物及探针 Primer and probe	引物及探针序列(5′→3′) Primer and probe sequence
aMPV-B-F	AATAGTCCTCAAGCAAGTCCTCAG
aMPV-B-R	CTGTTGTAATTTGACCTGTTCTACACT
aMPV-B-P	FAM-CTGGTGTTATCAGCCTTAGGCTTGACGCT-TAMRA

[1]	邬沛伶, 李依璇, 王浩杰, 李亚菲, 刘绍蒙, 刘青芸, 王湘如. 猪流行性腹泻疫苗研究进展[J]. 畜牧兽医学报, 2025, 56(3): 1042-1058.
[2]	于泽坤, 姜承源, 袁洪兴, 周生, 段笑笑, 李彦, 宋勤叶. B亚型禽偏肺病毒B1分离株感染性克隆的构建与鉴定[J]. 畜牧兽医学报, 2025, 56(2): 788-802.
[3]	李枝卉, 罗金林, 谢虹, 李明坤, 林杨, 张宇姣, 徐引弟, 陈焕春, 蔡旭旺, 徐晓娟. 副猪格拉瑟菌OppA过表达株的构建及其对小鼠免疫保护性研究[J]. 畜牧兽医学报, 2025, 56(2): 860-869.
[4]	曾焱, 欧祥龙, 闫晓阳, 刘灿, 廖永洪. 猪胸膜肺炎放线杆菌OmpD、LppB蛋白的原核表达及免疫原性分析[J]. 畜牧兽医学报, 2025, 56(1): 343-352.
[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.
[6]	伭婷, 杨凯一, 蔡金双, 耿琰, 李玉峰. 副猪格拉瑟菌6个候选抗原对小鼠的免疫特性分析[J]. 畜牧兽医学报, 2023, 54(5): 2073-2082.
[7]	孟令宅, 陈春丽, 于蒙蒙, 王占新, 王素艳, 刘鹏, 何塔娜, 郭茹, 陈运通, 刘长军, 祁小乐, 吴志强, 高玉龙. B亚型禽偏肺病毒对黄羽肉鸡致病性分析及其灭活疫苗免疫效果评价[J]. 畜牧兽医学报, 2023, 54(12): 5154-5161.
[8]	尹德晶, 吴燕, 岳筠, 杨鹏, 陈静, 王慧, 张双翔, 王开功, 程振涛. 丝状支原体山羊亚种LppA蛋白N末端基因真核表达载体构建及小鼠免疫效果分析[J]. 畜牧兽医学报, 2022, 53(3): 847-856.
[9]	于蒙蒙, 包媛玲, 王素艳, 辛子琪, 刘鹏, 冯笑艳, 孟令宅, 郭茹, 张艳萍, 刘长军, 祁小乐, 李俊平, 王笑梅, 高玉龙. B亚型禽偏肺病毒的分离鉴定及致病性研究[J]. 畜牧兽医学报, 2022, 53(10): 3540-3549.
[10]	马小静, 李梦莹, 张淮瑜, 宋阿北, 许立华, 吴心华, 张巧娥, 李继东. 共表达牛病毒性腹泻病毒E0基因和牛传染性鼻气管炎病毒gD基因的重组质粒DNA构建及其对小鼠的免疫效应[J]. 畜牧兽医学报, 2021, 52(1): 154-165.
[11]	孙普, 何伟, 付元芳, 李冬, 杨林, 魏德陇, 曹轶梅, 李平花, 白兴文, 马雪青, 李坤, 包慧芳, 张婧, 朱新荣, 刘在新, 卢曾军. 口蹄疫灭活疫苗的免疫效果及其对鉴别诊断的干扰[J]. 畜牧兽医学报, 2020, 51(10): 2481-2489.
[12]	陈晓, 葛雷, 戴建军, 李丽, 张德福, 吴彩凤, 张树山. 羊口疮病毒F1L、B2L基因重组腺病毒构建及对小鼠的免疫效果分析[J]. 畜牧兽医学报, 2018, 49(8): 1701-1708.
[13]	黄莉, 谢芝勋, 谢丽基, 黄娇玲, 谢志勤, 范晴, 邓显文, 罗思思, 张艳芳, 曾婷婷, 王盛, 张民秀. 禽呼肠孤病毒强毒株和弱毒疫苗株感染鸡肝癌细胞的转录组学分析[J]. 畜牧兽医学报, 2017, 48(11): 2135-2147.
[14]	任志浩，房立春，栾怀彪，李阳，王一新，崔治中，常爽，赵鹏. 鸡传染性贫血病毒荧光定量PCR检测方法的建立及其在疫苗污染检测中的应用[J]. 畜牧兽医学报, 2016, 47(7): 1459-1464.
[15]	黄莉，谢芝勋，谢丽基，邓显文，谢志勤，范晴，罗思思，黄娇玲，曾婷婷，张艳芳，王盛. 禽呼肠孤病毒强毒株和弱毒疫苗株感染Vero细胞的蛋白质组学研究[J]. 畜牧兽医学报, 2016, 47(2): 331-339.

B亚型禽偏肺病毒病弱毒疫苗对商品蛋鸡的免疫效果

Study on Immune Effect of Subtype B Attenuated Avian Metapneumovirus Disease Vaccine on Commercial Layer Chickens

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 28

相关文章 15

编辑推荐

Metrics

本文评价