引起种（蛋）鸭输卵管积液综合征的新发病毒—C型禽偏肺病毒

傅秋玲; 江南松; 梁齐章; 刘荣昌; 万春和; 程龙飞; 陈红梅; 林琳; 焦文龙; 吴胜会; 江斌; 郑小兰; 林建生; 傅光华; 黄瑜

doi:10.19303/j.issn.1008-0384.2023.12.001

引起种（蛋）鸭输卵管积液综合征的新发病毒—C型禽偏肺病毒

doi: 10.19303/j.issn.1008-0384.2023.12.001

1.
福建省农业科学院畜牧兽医研究所/福建省禽病防治重点实验室/福建省畜禽疫病防控产业技术创新研究院，福建　福州　350013

基金项目: 新世纪“百千万人才工程”国家级人选科研补助基金（NCNCMTPC-2009）；国家现代农业（水禽）产业技术体系建设专项（CARS-42）；福建省科技计划公益类专项（2022R10260011）；福建省农业科学院青年英才项目（YC2019010）；福建省农业科学院科技创新团队建设项目（CXTD2021005-1）

详细信息

作者简介:
傅秋玲（1985 —），女，博士，助理研究员，主要从事水禽病毒诊断与防控研究，E-mail：qiulingfu0822@163.com

通讯作者:
傅光华（1977 —），男，博士，研究员，主要从事畜禽传染病研究，E-mail：fuyuan163@163.com

黄瑜（1965 —），男，博士，研究员，主要从事畜禽传染病研究，E-mail：huangyu_815@163.com

中图分类号: S855.3
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出版历程
- 收稿日期: 2023-10-16
- 修回日期: 2023-12-15
- 网络出版日期: 2024-01-25
- 刊出日期: 2023-12-28

Hydrosalpinx Fluid Syndrome in Ducks Caused by Avian Metapneumovirus Subgroup C

1.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural sciences/Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention/Fujian Industry Technology Innovation Research Academy of Livestock and Poultry Diseases Prevention & Control, Fuzhou, Fujian　350013, China

摘要

摘要: 目的明确引起种（蛋）鸭特征性输卵管积液和产蛋下降的新发疫病[俗称种（蛋）鸭输卵管积液综合征]的病原。方法对自闽粤赣浙皖桂等地产蛋率较高的部分鸭群表现以输卵管内乳糜性积液、黏膜出血水肿增厚及不同程度产蛋下降为特征的病（死）麻鸭、樱桃谷种鸭和种番鸭中采集的样品（输卵管积液、输卵管黏膜、卵巢和肝脾脏器），开展鸭已知病原核酸监测、病原分离鉴定和开产麻鸭人工感染试验。结果细菌分离显示，输卵管积液里未分离到细菌，排除细菌感染引起；经病毒核酸监测，只有C型禽偏肺病毒（Avain metapneumoviurs subgroup C，aMPV/C）阳性，未检测到其他引起禽产蛋异常的病毒；以aMPV/C单一阳性的样品经鸭胚进行病毒分离与传代，发现接种鸭胚未见死亡，但胚体表面稍水肿和出血、肝脏出血；对第5代鸭胚液进行RT-PCR检测，结果显示aMPV/C分离阳性率为72.2%；对获得的4株分离株（JX2126、FJ2228、FJ2375和GD2381株）的基因片段进行测序和序列分析发现，该片段与最早报道的番鸭aMPV/C法国分离株（Muscovy duck/1999/99178/France）同源性最高，为96.1%，而与A、B、D型禽偏肺病毒（Avain metapneumoviurs subgroup A, B, D，aMPV/A, B, D）分离株的同源性仅分别为67.8%～68.7%、66.1%～66.8%和71.4%，与人偏肺病毒（Human metapneumoviurs，hMPV）分离株的同源性为67.7%～68.3%；遗传进化分析表明，以上4株分离株与aMPV/C处于同一进化分支上，而与其他亚型aMPV和hMPV亲缘关系较远；经开产麻鸭人工感染试验，成功复制出与自然感染病例相同的临床病症，并能回收到病毒。结论明确引起种（蛋）鸭输卵管积液综合征的病原为C型禽偏肺病毒。本研究为该病的快速准确诊断和精准防控提供科学依据。
- 种（蛋）鸭 /
- 输卵管积液综合征 /
- 产蛋下降 /
- 输卵管积液 /
- C型禽偏肺病毒 /
- 分离与鉴定
Abstract: Objective Pathogen causing the hydrosalpinx fluid syndrome (HFS) with a declined egg-laying in breeding and egg-laying ducks was investigated for the disease control. Method Nucleic acid amplification, pathogen isolation and identification, and artificial infection on ducks were carried out on tissue samples collected from the live or dead Sheldrake Ducks, Cherry Valley Ducks, and Muscovy Ducks that were found infected by HFS at farms in Fujian, Guangdong, Jiangxi, Zhejiang, Anhui, and Guangxi autonomous region. The disease symptoms included chylous hydrosalpinx fluids and hemorrhagic, edematous, and thickened mucosa, in addition to a declined egg-laying on the ducks. Result No bacteria were detected in the hydrosalpinx fluid. In the clinical specimens, RT-PCR tested positive on avian metapneumovirus subgroup C (aMPV/C) but no other commonly known viruses that cause a declined egg-laying in ducks. Subsequently, the duck embryos inoculated with the viral infected samples were aMPV/C-positive and showed slight embryonic edema and bleeding on the embryo surface and liver. The virus isolation rate on the F5 embryo fluid by RT-PCR was 72.2%. The nucleotide sequences of the isolated FJ2375, GD2381, FJ2228, and JX2126 had a homology of 96.1% with the earliest reported aMPV/C-infected Muscovy duck/1999/99178/France, those of 67.8-68.7%, 66.1-66.8%, and 71.4% with aMPV/A, B, and D, respectively, and that of 67.7-68.3% with the human MPV. The sequences were more closely related to aMPV/C than the others in genetic evolution. Furthermore, the virus was successfully recovered from the artificially infected ducks that exhibited similar HFS symptoms observed in naturally diseased birds. Conclusion The pathogenic virus that caused HFS in ducks was identified as aMPV/C. It paved the way for the development of means to accurately diagnose and effectively prevent and control the disease.
- breeding or egg-laying duck /
- hydrosalpinx fluid syndrome /
- declined egg-laying /
- hydrosalpinx fluid /
- Avian metapneumovirus subgroup C /
- isolation and identification

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图 1 输卵管内乳糜性积液

Figure 1. Chylous hydrosalpinx fluid

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图 2 输卵管黏膜出血水肿增厚

Figure 2. Hemorrhagic, edematous, and thickened mucosa in duck fallopian tubes

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图 3 临床样品中C型禽偏肺病毒（RT-）PCR检测结果

M为DL2000 maker，1～4分别为输卵管积液、输卵管黏膜、卵巢、肝脾脏器中C型禽偏肺病毒，5～14分别为禽流感病毒、禽坦布苏病毒、鸭减蛋综合征病毒、鸭瘟病毒、禽副黏病毒、鸭星状病毒3型、巴泰病毒、鸭1型甲肝病毒、A型禽偏肺病毒和B型禽偏肺病毒，15为空白对照。

Figure 3. RT-PCR detection of aMPV/C in clinical samples

M: DL2000 maker; 1-4: hydrosalpinx fluid, hydrosalpinx mucosa, ovarium, liver, and spleen, respectively; 5-14: AIV, ATV, EDS, DPV, APMV, DAstV-3, BTV, DHAV-1, aMPV/A and aMPV/B, respectively; 15: control.

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图 4 胚体稍水肿和表面出血

左边胚体为接毒组，右边胚体为对照组。

Figure 4. Slight edema and surface bleeding on embryo

Left: infected embryos; right: control.

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图 5 分离株C型禽偏肺病毒RT-PCR检测结果

M为DL2000 maker，1～5、7、8、10、14～18为aMPV/C阳性，6、9、11～13为aMPV/C阴性，19为空白对照。

Figure 5. RT-PCR analysis on isolated aMPV/C

M: DL2000 maker; 1-5, 7, 8, 10, and 14-18: aMPV/C positive; 6, 9, 11-13: aMPV/C negative; 19: control.

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图 6 鸭C型禽偏肺病毒分离株基因片段的遗传进化树

Figure 6. Phylogenetic tree of aMPV/C isolates from ducks

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表 1 麻鸭感染试验结果

Table 1. Results of infection test on Sheldrake Ducks

组别 Groups	数量 Number /n	攻毒前产蛋数和产蛋率 Eggs and laying rate before Infection/（n/%）	攻毒后14 d内产蛋数和产蛋率 Eggs and laying rate during 14 days post infectious/（n/%）										攻毒后总产蛋数和平均产蛋率 Total eggs and the mean of laying rate during 14 days post infectious/（n/%）
组别 Groups	数量 Number /n	攻毒前产蛋数和产蛋率 Eggs and laying rate before Infection/（n/%）	1	2	3	4	5～9	10	11	12	13	14
攻毒组 Infection group	25	22/88	18/72	18/72	15/60	11/44	8/32	9/36	10/40	9/36	10/40	11/44	151/43.1
对照组 Control group	25	22/88	19/76	18/72	17/68	18/72	19/76	20/80	21/84	20/80	21/84	21/84	270/77.1

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