鸭短喙与侏儒综合征病毒分离与鉴定

doi:10.11843/j.issn.0366-6964.2024.08.033

摘要/Abstract

摘要：

山东某樱桃谷鸭场出现以患鸭短喙、长舌、生长不良为主要特征的疾病，疑似感染鸭短喙与侏儒综合征病毒(duckling short beak and dwarfism syndrome virus, SBDSV)。本研究旨在分离鉴定该病毒，并检测其对樱桃谷鸭的致病性。首先，通过临床症状、病理变化、病料PCR检测，确定感染鸭短喙侏儒综合征病毒。其次，用SPF鸭胚分离培养病毒后，对病毒进行电镜观察和同源性与系统发育树分析，再检测病毒尿囊液EID₅₀。最后，动物回归试验中，给2日龄樱桃谷鸭口服病毒感染鸭胚尿囊液，观察致病特点，以及用PCR和间接免疫荧光试验检测病毒分布。结果显示：SPF鸭胚接毒后96~120 h鸭胚死亡，电镜观察病毒粒子大小为20~50 nm，尿囊液EID₅₀为10^-4.85·0.2 mL^-1。尿囊液PCR检测SBDSV为阳性，证明从鸭胚中成功分离培养SBDSV。该病例主要临床表现短喙和体重降低，感染后2周内持续排毒，VP2片段与鸭短喙侏儒综合征病毒序列的相似性为99.8%，表明是由鸭短喙与侏儒综合征病毒感染引起，命名为SBDS-SD株。动物回归试验中，出现短喙、腹泻、体重显著降低等与自然病例相似的临床症状。本试验成功分离到一株鸭短喙与侏儒综合征病毒，并验证其对樱桃谷鸭具有致病性，为该病的进一步研究提供基础。

关键词: 鸭短喙与侏儒综合征, 致病性试验, 病毒增殖培养, 电镜观察

Abstract:

A disease characterized by short beak, long tongue, and poor growth was observed in a cherry valley duck farm in Shandong, suspected to be infected with short beak and dwarfism syndrome virus (SBDSV). This study aimed to isolate and identify the virus, and to test its pathogenicity to cherry valley ducks. First, the infection of short beak and dwarfism syndrome virus was confirmed by clinical symptoms, pathological changes, and PCR detection of the diseased materials. Second, after isolating and culturing the virus in SPF duck embryos, it was observed by electron microscopy and analyzed by homology and phylogenetic tree, and then the EID₅₀ of the virus allantoic fluid was tested. Finally, in the animal reversion test, 2-day-old cherry valley ducks were orally administered with allantoic fluid, and the pathogenic characteristics were observed, as well as the virus distribution was detected by PCR and indirect immunofluorescence assay. The results showed that SPF duck embryos died at 96-120 hours post-inoculation, and virus particles of 20-50 nm in size were observed by electron microscopy. The allantoic fluid had an EID₅₀ of 10^-4.85·0.2 mL^-1. The allantoic fluid was positive for SBDSV by PCR, confirming the successful isolation and cultivation of SBDSV from duck embryos. The main clinical manifestations of this case were short beak and weight loss. The virus was shed continuously for two weeks after infection. The VP2 fragment had a 99.8% similarity with the sequence of duck short beak and dwarfism syndrome virus, indicating that it was caused by the infection of this virus, which was named SBDS-SD strain. In the animal challenge experiment, clinical symptoms similar to those of the natural case, such as short beak, diarrhea and significant weight loss, were observed. This experiment successfully isolated a strain of short beak and dwarfism syndrome virus and verified its pathogenicity to cherry valley ducks, providing a basis for further research on this disease.

Key words: duck short beak and dwarf syndrome, pathogenicity test, virus proliferation culture, electron microscope observation

中图分类号:

S852.659.2

孙雨点, 宋紫玥, 张洪亮, 秦志华, 单虎, 杨瑞梅. 鸭短喙与侏儒综合征病毒分离与鉴定[J]. 畜牧兽医学报, 2024, 55(8): 3623-3630.

Yudian SUN, Ziyue SONG, Hongliang ZHANG, Zhihua QIN, Hu SHAN, Ruimei YANG. Isolation and ldentification of Duckling Short Beak and Dwarfism Syndrome Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3623-3630.

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组别 Group	时间 Time	攻毒组 Attack group	对照组 Control group
体重/g	第14天Day 14	147.22±44.05^*	215.00±23.20
Body weight	第21天Day 21	179.77±48.00^**	280.25±21.12
	第35天Day 35	277.14±60.61^**	413.66±20.22
喙长/mm	第14天Day 14	23.15±1.75^*	25.39±0.86
Beak length	第21天Day 21	25.06±2.85^*	28.39±0.88
	第35天Day 35	28.71±3.64^*	33.82±1.05

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