传染性皮下及造血组织坏死病毒(IHHNV)在双壳贝类中的感染情况研究
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摘要
采用国际兽医局推荐的传染性皮下及造血组织坏死病毒(infectious hypodermal and hematopoietic necrosis virus,IHHNV)检测方法,首次开展IHHNV在双壳贝类(中国蛤蜊、泥蚶、花蛤、缢蛏、文蛤和白蛤)和螺类软体动物(螺蛳、中华圆田螺)中的感染情况。此外,本研究以泥蚶为研究对象,建立了IHHNV在双壳贝类中的实验室感染模型,研究了人工感染泥蚶后IHHNV在其体内的增殖和致病情况。结果显示:在采集的不同种类贝类样品中均检测到IHHNV,其中在泥蚶中的阳性率最高(50%),在文蛤中的阳性率最低(15%),表明IHHNV在双壳贝类中的分布较为广泛,是IHHNV的重要携带物种。在采集的螺类软体动物中均未检测到IHHNV。系统发育分析表明,来自不同贝类中的IHHNV构成了进化树中不同的分支,其中中国蛤蜊、花蛤、白蛤源的IHHNV属于Ⅱ型感染株,而泥蚶、缢蛏、文蛤源的IHHNV单独成簇,形成了一个全新的分支。实验室感染结果显示,IHHNV对泥蚶没有明显的致病性,但可在其体内增殖,在感染后第5天其体内的病毒载量达到最大(1.8×104copies/g),随着感染时间的增加,其体内病毒含量呈现递减趋势,但在感染后第30天体内仍可检测到病毒的存在。本研究结果对对虾养殖尤其是虾贝混养模式中预防和控制IHHNV的传播具有重要意义。
Detection to the infectious hypodermal and hematopoietic necrosis virus(IHHNV) in bivalve shellfish(Mactra chinensis, Tegillarca granosa, Ruditapes philippinarum, Sinonovacula constricta, Meretrix meretrix and Mactra veneriformis) and snails(Margarya melanioides and Cipangopaludina cahayensis) were first attempted using the method that recommended by the OIE(Office International des Epizooties). We regarded T. granosa as the research object to establish the IHHNV infection model in bivalve shellfish and studied the replication and pathogenicity of IHHNV in bivalve shellfish after infection. Results demonstrated that IHHNV could be detected in all kinds of the collected shellfish samples with the highest positive rate(50%) in T. granosa and the lowest(15%) in M. meretrix, indicating that IHHNV widely distributed in bivalve shellfish, and bivalve shellfish is an important carrier of IHHNV. However, no IHHNV was detected in the collected snails. Phylogenetic tree analysis indicated that IHHNV from different shellfish species belong to different branches. Among them, IHHNV from M. chinensis, R. philippinarum and M. veneriformis belonged to type II, while IHHNV from T. granosa, S. constricta and M. meretrix formed a completely new branch. The result of infection showed that IHHNV could replicate in T. granosa with no obvious pathogenicity. The maximum viral load in T. granosa(1.8×104 copies/g) reached the peak value 5 days post infection. The viral load showed a decreasing trend with the increase of infection time, but the virus could be also detected until 30 days post infection. Therefore, the results of this study are of great significance for the prevention and control of IHHNV transmission in shrimp farming, especially in shrimp and shellfish co-culture farming.
Detection to the infectious hypodermal and hematopoietic necrosis virus(IHHNV) in bivalve shellfish(Mactra chinensis, Tegillarca granosa, Ruditapes philippinarum, Sinonovacula constricta, Meretrix meretrix and Mactra veneriformis) and snails(Margarya melanioides and Cipangopaludina cahayensis) were first attempted using the method that recommended by the OIE(Office International des Epizooties). We regarded T. granosa as the research object to establish the IHHNV infection model in bivalve shellfish and studied the replication and pathogenicity of IHHNV in bivalve shellfish after infection. Results demonstrated that IHHNV could be detected in all kinds of the collected shellfish samples with the highest positive rate(50%) in T. granosa and the lowest(15%) in M. meretrix, indicating that IHHNV widely distributed in bivalve shellfish, and bivalve shellfish is an important carrier of IHHNV. However, no IHHNV was detected in the collected snails. Phylogenetic tree analysis indicated that IHHNV from different shellfish species belong to different branches. Among them, IHHNV from M. chinensis, R. philippinarum and M. veneriformis belonged to type II, while IHHNV from T. granosa, S. constricta and M. meretrix formed a completely new branch. The result of infection showed that IHHNV could replicate in T. granosa with no obvious pathogenicity. The maximum viral load in T. granosa(1.8×104 copies/g) reached the peak value 5 days post infection. The viral load showed a decreasing trend with the increase of infection time, but the virus could be also detected until 30 days post infection. Therefore, the results of this study are of great significance for the prevention and control of IHHNV transmission in shrimp farming, especially in shrimp and shellfish co-culture farming.
引文
白丽蓉,赵志英,2012.对虾传染性皮下与造血组织坏死病毒(IHHNV)的研究进展.中国农学通报,28(14):114-119
陈萍,王芳芳,翁歆之,2016.海水围塘虾蟹贝生态混养模式研究.中国水产,(11):109-112
范东东,魏永伟,苗亮等,2015.罗氏沼虾(Macrobrachium rosenbergii)传染性皮下和造血器官坏死病毒(IHHNV)的流行病学调查.海洋与湖沼,46(5):1153-1159
焦永真,韩剑秋,王宪明等,1990.1988年上海甲型肝炎暴发流行中从毛蚶分离到甲型肝炎病毒.病毒学报,6(4):312-315
Browdy C L,Holloway Jr J D,King C O et al,1993.IHHN virus and intensive culture of Penaeus vannamei:effects of stocking density and water exchange rates.Journal of Crustacean Biology,13(1):87-94
Campos C J A,Lees D N,2014.Environmental transmission of human noroviruses in shellfish waters.Applied and Environmental Microbiology,80(12):3552-3561
Chai C,Liu Y C,Xia X M et al,2016.Prevalence and genomic analysis of infectious hypodermal and hematopoietic necrosis virus(IHHNV)in Litopenaeus vannamei shrimp farmed in Shanghai,China.Archives of Virology,161(11):3189-3201
Dhar A K,Roux M M,Klimpel K R,2001.Detection and quantification of infectious hypodermal and hematopoietic necrosis virus and white spot virus in shrimp using Real-Time quantitative PCR and SYBR green chemistry.Journal of Clinical Microbiology,39(8):2835-2845
Encinas-García T,Mendoza-Cano F,Enríquez-Espinoza T et al,2015.An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.Journal of Virological Methods,212:53-58
Galván-Alvarez D,Mendoza-Cano F,Hernández-López J et al,2012.Experimental evidence of metabolic disturbance in the white shrimp Penaeus vannamei induced by the infectious hypodermal and hematopoietic necrosis virus(IHHNV).Journal of Invertebrate Pathology,111(1):60-67
Gao S Y,Li D D,Zha E H et al,2015.Surveillance of hepatitis Evirus contamination in shellfish in China.International Journal of Environmental Research and Public Health,12(2):2026-2036
Kalagayan H,Godin D,Kanna R et al,1991.IHHN virus as an etiological factor in runt-deformity syndrome(RDS)of juvenile Penaeus vannamei cultured in Hawaii.Journal of the World Aquaculture Society,22(4):235-243
Kim J H,Kim H K,Nguyen V G et al,2012.Genomic sequence of infectious hypodermal and hematopoietic necrosis virus(IHHNV)KLV-2010-01 originating from the first Korean outbreak in cultured Litopenaeus vannamei.Archives of Virology,157(2):369-373
Lees D,2000.Viruses and bivalve shellfish.International Journal of Food Microbiology,59(1-2):81-116
Lightner D V,2011.Virus diseases of farmed shrimp in the Western Hemisphere(the Americas):a review.Journal of Invertebrate Pathology,106(1):110-130
Lightner D V,Redman R M,Bell T A et al,1983.Detection of IHHN virus in Penaeus stylirostris and P.vannamei imported into Hawaii.Journal of the World Aquaculture Society,14(1-4):212-225
Rai P,Safeena M P,Krabsetsve K et al,2012.Genomics,molecular epidemiology and diagnostics of infectious hypodermal and hematopoietic necrosis virus.Indian Journal of Virology,23(2):203-214
Silva D C D,Nunes A R D,Teixeira D I A et al,2014.Infectious hypodermal and hematopoietic necrosis virus from Brazil:sequencing,comparative analysis and PCR detection.Virus Research,189:136-146
Vega-Heredia S,Mendoza-Cano F,Sánchez-Paz A,2012.The infectious hypodermal and haematopoietic necrosis virus:a brief review of what we do and do not know.Transboundary and Emerging Diseases,59(2):95-105
Wei Y W,Fan D D,Chen J,2017.The mussel Mytilus edulis L.as an important reservoir of infectious hypodermal and hematopoietic necrosis virus(IHHNV).Aquaculture Research,48(3):1346--1350
陈萍,王芳芳,翁歆之,2016.海水围塘虾蟹贝生态混养模式研究.中国水产,(11):109-112
范东东,魏永伟,苗亮等,2015.罗氏沼虾(Macrobrachium rosenbergii)传染性皮下和造血器官坏死病毒(IHHNV)的流行病学调查.海洋与湖沼,46(5):1153-1159
焦永真,韩剑秋,王宪明等,1990.1988年上海甲型肝炎暴发流行中从毛蚶分离到甲型肝炎病毒.病毒学报,6(4):312-315
Browdy C L,Holloway Jr J D,King C O et al,1993.IHHN virus and intensive culture of Penaeus vannamei:effects of stocking density and water exchange rates.Journal of Crustacean Biology,13(1):87-94
Campos C J A,Lees D N,2014.Environmental transmission of human noroviruses in shellfish waters.Applied and Environmental Microbiology,80(12):3552-3561
Chai C,Liu Y C,Xia X M et al,2016.Prevalence and genomic analysis of infectious hypodermal and hematopoietic necrosis virus(IHHNV)in Litopenaeus vannamei shrimp farmed in Shanghai,China.Archives of Virology,161(11):3189-3201
Dhar A K,Roux M M,Klimpel K R,2001.Detection and quantification of infectious hypodermal and hematopoietic necrosis virus and white spot virus in shrimp using Real-Time quantitative PCR and SYBR green chemistry.Journal of Clinical Microbiology,39(8):2835-2845
Encinas-García T,Mendoza-Cano F,Enríquez-Espinoza T et al,2015.An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.Journal of Virological Methods,212:53-58
Galván-Alvarez D,Mendoza-Cano F,Hernández-López J et al,2012.Experimental evidence of metabolic disturbance in the white shrimp Penaeus vannamei induced by the infectious hypodermal and hematopoietic necrosis virus(IHHNV).Journal of Invertebrate Pathology,111(1):60-67
Gao S Y,Li D D,Zha E H et al,2015.Surveillance of hepatitis Evirus contamination in shellfish in China.International Journal of Environmental Research and Public Health,12(2):2026-2036
Kalagayan H,Godin D,Kanna R et al,1991.IHHN virus as an etiological factor in runt-deformity syndrome(RDS)of juvenile Penaeus vannamei cultured in Hawaii.Journal of the World Aquaculture Society,22(4):235-243
Kim J H,Kim H K,Nguyen V G et al,2012.Genomic sequence of infectious hypodermal and hematopoietic necrosis virus(IHHNV)KLV-2010-01 originating from the first Korean outbreak in cultured Litopenaeus vannamei.Archives of Virology,157(2):369-373
Lees D,2000.Viruses and bivalve shellfish.International Journal of Food Microbiology,59(1-2):81-116
Lightner D V,2011.Virus diseases of farmed shrimp in the Western Hemisphere(the Americas):a review.Journal of Invertebrate Pathology,106(1):110-130
Lightner D V,Redman R M,Bell T A et al,1983.Detection of IHHN virus in Penaeus stylirostris and P.vannamei imported into Hawaii.Journal of the World Aquaculture Society,14(1-4):212-225
Rai P,Safeena M P,Krabsetsve K et al,2012.Genomics,molecular epidemiology and diagnostics of infectious hypodermal and hematopoietic necrosis virus.Indian Journal of Virology,23(2):203-214
Silva D C D,Nunes A R D,Teixeira D I A et al,2014.Infectious hypodermal and hematopoietic necrosis virus from Brazil:sequencing,comparative analysis and PCR detection.Virus Research,189:136-146
Vega-Heredia S,Mendoza-Cano F,Sánchez-Paz A,2012.The infectious hypodermal and haematopoietic necrosis virus:a brief review of what we do and do not know.Transboundary and Emerging Diseases,59(2):95-105
Wei Y W,Fan D D,Chen J,2017.The mussel Mytilus edulis L.as an important reservoir of infectious hypodermal and hematopoietic necrosis virus(IHHNV).Aquaculture Research,48(3):1346--1350