行动障碍野田村病毒(MDNV)自然感染贝类的初步研究
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中国水产科学研究院基本科研业务费(2020TD39; 2023TD42)、中国水产科学研究院黄海水产研究所级基本科研业务费(20603022024023)和国家虾蟹产业技术体系(CARS-48)共同资助


Preliminary study on natural infection of shellfish with movement disorder nodavirus (MDNV)
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    摘要:

    行动障碍野田村病毒(movement disorder nodavirus, MDNV)是α野田村病毒属的一类新型野田村病毒。为了解MDNV在贝类中的自然感染情况,本研究综合运用TaqMan实时荧光定量PCR (TaqMan RT-qPCR)、组织病理和组织原位杂交等检测手段,对黄海海区野生贝类样品携带和感染MDNV的情况进行了系统分析。TaqMan RT-qPCR检测结果显示,所采集的太平洋牡蛎(Crossostrea gigas)和栉孔扇贝(Azumapecten farreri)中均有MDNV阳性检出,前者的阳性检出率为16.67% (1/6),后者的阳性检出率为33.3% (2/6);在魁蚶(Scapharca broughtonii)和菲律宾蛤仔(Ruditapes philippinarum)中均未检测到MDNV。组织病理分析显示,太平洋牡蛎外套膜组织中存在大量的嗜酸性包涵体,细胞核发生固缩;鳃组织上皮细胞受损,胞质内出现空泡化,部分细胞核固缩,鳃丝内充斥着大量的嗜酸性包涵体。栉孔扇贝消化腺上皮细胞的细胞核内可见异常嗜酸性包涵体,鳃组织中部分上皮细胞的细胞核发生核固缩。原位杂交结果显示,太平洋牡蛎和栉孔扇贝的病变部位均可见深紫色或淡紫色的MDNV探针杂交信号。来源于贝类的MDNV、虾类的MDNV和偷死野田村病毒(CMNV)的RNA聚合酶(RdRp)序列比对分析显示,太平洋牡蛎与栉孔扇贝的MDNV序列相似度为99.3%,而这二者与CMNV原始分离株序列片段的相似度为83.3%。本研究首次证实,MDNV在自然条件下能够感染近海区域的野生贝类,并导致靶组织发生明显的病理损伤,同时也表明,需密切关注患病贝类作为传染源或宿主导致MDNV在水产养殖动物中大规模传播和流行的风险。

    Abstract:

    China's marine aquaculture is rich and diverse. As China's marine aquaculture industry continues to expand, the impact of disease problems on this sector has become increasingly prominent, leading to substantial economic losses for the country's aquaculture enterprises. Shellfish is an important source of high-quality protein for human beings and simultaneously plays an important ecological service function in the marine ecosystem. Shellfish aquaculture in China mainly focuses on marine shellfish, accounting for the highest proportion of the total marine aquaculture output in China. Recently, pathogenic infections, including bacteria, fungi, and parasites, have caused notable disease outbreaks in China's mariculture shellfish. For example, oyster herpesvirus and its variant viruses can cause disease infections in many shellfish organisms and are important pathogens that lead to shellfish mortality. Additionally, shellfish farming in China's northern Yellow Sea area is often infected by Vibrio canis and Bacillus luminescens. Regarding the developmental changes of diseases, shellfish diseases in mariculture mainly include pathogenic and non-pathogenic diseases. Pathogenic diseases have harmful manifestations, such as bacteria, parasites, fungi, and pathogenic worms, whereas non-pathogenic diseases are mainly caused by mechanical injuries, drug abuse, and physical or chemical stimuli. Shellfish diseases in mariculture often involve multiple infections and cross-contamination, and non-pathogenic diseases can sometimes develop into pathogenic hazards. Around 2003, "viral covert mortality disease (VCMD)" appeared in the Penaeus vannamei culture area in southern China. VCMD is an emerging infectious disease caused by covert mortality nodavirus (CMNV). The cross-species transmission capacity of CMNV has caused huge economic losses to the aquaculture industry. Movement disorder nodavirus (MDNV) is similar to CMNV, both of which are members of the newly discovered α-nodavirus genus. P. vannamei, infected with MDNV, showed sinking characteristics and decreased swimming performance. To understand whether MDNV has the same cross-species transmission ability as CMNV, we collected wild shellfish samples in the offshore area of the Yellow Sea and the East China Sea and systematically analyzed the possibility of the shellfish carrying and infecting by MDNV. TaqMan real-time fluorescence quantitative PCR (TaqMan RT-qPCR), histopathology, and in situ hybridization were used in this investigation. The results of the TaqMan RT-qPCR assay showed that MDNV was detected in Crossostrea gigas and Azumapecten farreri collected in the offshore area of the Yellow Sea and East China Sea. The positive detection rates of MDNV in C. gigas and A. farreri was 16.67% (1/6) and 33.3% (2/6), respectively. MDNV was not detected in Scapharca broughtonii and Ruditapes philippinarum. Histopathology and in situ hybridization analysis showed many eosinophilic inclusions in the mantle tissue of C. gigas, and nuclear pyknosis occurred. In the gill tissue of C. gigas, epithelial cells were severely damaged, and the appearance of vacuolation and necrosis of karyopyknotic nuclei was observed in the gill. The gill filaments were filled with many eosinophilic inclusions. The epithelial cells of A. farreri chlamys were filled with eosinophilic substances, and the karyopyknotic nuclei of some epithelial cells were also observed. The positive hybridization signals of the purple MDNV RNA probe were observed in the lesion sites of C. gigas and A. farreri. In addition, the MDNV RNA-dependent RNA polymerase nucleic acid sequences were cloned and then submitted to NCBI web multi-sequence alignments using the online program. The multiple sequence alignment results showed that the similarity of MDNV sequence fragments of C. gigas and A. farreri was 99.3%, and the similarity between them and the original CMNV isolates was 83.3%. This study shows that MDNV can cross the species barrier under natural conditions, infect wild shellfish in the offshore area of the Yellow Sea, and cause evident pathological damage to target tissues. It also suggests that attention should focus on avoiding the use of MDNV-infected shellfish seed in aquaculture operations to prevent large-scale transmission or epidemic of MDNV in cultured shellfish.

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李萍,夏继涛,白昌明,徐瑞东,徐婷婷,刘爽,姚亮,余星潼,张庆利.行动障碍野田村病毒(MDNV)自然感染贝类的初步研究.渔业科学进展,2025,46(4):217-229

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  • 收稿日期:2024-08-23
  • 最后修改日期:2024-10-05
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  • 在线发布日期: 2025-07-14
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