鲤浮肿病毒人工感染锦鲤的试验研究
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摘要
为研究鲤浮肿病毒的感染方式及在锦鲤各组织器官中的动态分布,在水温20～22℃,溶解氧7～8mg/L试验条件下,采用注射、浸泡、划伤后浸泡3种方式人工感染锦鲤,观察了试验鱼的发病和死亡情况;检测了鲤浮肿病毒在感染鱼体内各组织器官中的存在情况,探讨了主要被感染组织的病理变化。试验结果显示,采用注射、浸泡、划伤后浸泡3种方式均可导致健康锦鲤感染该病毒,病鱼出现昏睡、烂鳃、凹眼等症状,累积死亡率35%～88%。锦鲤感染该病毒后至少有3d的潜伏期,随后病毒在鳃组织中大量增殖,至7d达到高峰,病毒量314.8个/ng,之后病毒量开始逐渐下降,至12d在鳃组织中检测不出病毒。鲤浮肿病毒在肾、肝、脑、肌肉和脾中存在的时间较短且病毒量较低,在血液和肠道中未检出。结合鳃组织病理切片可知,鳃为该病毒感染的主要靶器官。本研究结果为鲤浮肿病的诊断和防控提供了一定的科学依据。
To study the artificial infection of Carp edema virus(CEV)and its dynamic distribution in different organs,artificial infecting tests were carried out by injection,by bath with and without scarifiction on skin using filtrated suspension of gills and kidney homogenates obtained from sick fish.The experimental water conditions were kept at temperature of 20—22 ℃ and at dissolved oxygen level of 7—8mg/L.Morbidity and mortality were observed and CEV were examined in gill,hepatopancreas,kidney,spleen,brain,blood,intestine and muscle by real-time PCR.The histopathological changes in the gills of infected koi Cyprinus carpio koi were assayed also.The results showed that CEV infected healthy koi by injection or by bath,especially by bath with scarification.And the sick koi showed typical clinical signs of CEVD including lethargy,rot gills and enophthalmos,with cumulative mortality of 35%—88%.In addition,CEV had a latent period of 3das post-infection at least,proliferated in gill tissue subsequently and reached its peak at 7dpost-infection as the virus loaded about 314.8copies/ng tissues.Then virus loads began to decline gradually,and were not be detected at 12dpost-infection.By contrast,CEV showed shorter multiplication time and lower virus loads in kidney,hepatopancreas,brain,muscle and spleen,without in blood and intestine.So combined with the histopathological observation,it was speculated that gill was one of the main target organs for CEV.The findings will provide a scientific basis for diagnosis,prevention and control of CEVD.
To study the artificial infection of Carp edema virus(CEV)and its dynamic distribution in different organs,artificial infecting tests were carried out by injection,by bath with and without scarifiction on skin using filtrated suspension of gills and kidney homogenates obtained from sick fish.The experimental water conditions were kept at temperature of 20—22 ℃ and at dissolved oxygen level of 7—8mg/L.Morbidity and mortality were observed and CEV were examined in gill,hepatopancreas,kidney,spleen,brain,blood,intestine and muscle by real-time PCR.The histopathological changes in the gills of infected koi Cyprinus carpio koi were assayed also.The results showed that CEV infected healthy koi by injection or by bath,especially by bath with scarification.And the sick koi showed typical clinical signs of CEVD including lethargy,rot gills and enophthalmos,with cumulative mortality of 35%—88%.In addition,CEV had a latent period of 3das post-infection at least,proliferated in gill tissue subsequently and reached its peak at 7dpost-infection as the virus loaded about 314.8copies/ng tissues.Then virus loads began to decline gradually,and were not be detected at 12dpost-infection.By contrast,CEV showed shorter multiplication time and lower virus loads in kidney,hepatopancreas,brain,muscle and spleen,without in blood and intestine.So combined with the histopathological observation,it was speculated that gill was one of the main target organs for CEV.The findings will provide a scientific basis for diagnosis,prevention and control of CEVD.
引文
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[6] Way K,Stone D.Emergence of carp edema virus-like(CEV-like)disease in the UK[J].CEFAS Finfish News,2013(15):32-34.
[7] Haenen O,Way K,Stone D,et al.“Koi Sleepy Disease”voor het eerst in Nederland aanqetoond in koikarpers[J].Tijdschr Dierqeneeskd,2014(4):26-29.
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[9] Lewisch E,Gorgoglione B,Way K,et al.Carp edema virus/kio sleepy disease:an emerging disease in central-east Europe[J].Transboundary and Emerging Diseases,2014,62(1):6-12.
[10]Jung-Schroers V,Adamek M,Teitge F,et al.Another potential carp killer carp edema virus disease in Germany[J].BMC Veterinary Research,2015,11(1):114-117.
[11]徐立蒲,王小亮,张文,等.养殖锦鲤鱼浮肿病的检测和鉴定[J].中国畜牧兽医,2017,44(2):613-618.
[12]徐立蒲,王静波,张文,等.鲤鱼浮肿病流行情况的初步研究[J].检验检疫学刊,2016,26(5):14-16.
[13]王小亮,徐立蒲,王姝,等.养殖鲤和锦鲤中的鲤浮肿病毒检测与分析[J].中国畜牧兽医,2017,44(10):213-218.
[14]温智清,刘莹,唐绍林,等.云南锦鲤养殖场鲤浮肿病毒的鉴定及基因型分析[J].病毒学报,2017,33(6):1-9.
[15]Zhang X,Ni Y,Ye J.Carp edema virus,an emerging threat to the carp(Cyprinus carpio)industry in China[J].Aquac Res,2017,474(1):34-39.
[16]吕晓楠,徐立蒲,王姝,等.鲤浮肿病研究进展[J].中国动物检疫,2018,35(5):75-80.
[17]徐立蒲,张文,王姝,等.河南沿黄滩区鲤浮肿病和锦鲤疱疹病毒病初步调查研究[J].科学养鱼,2017(9):59-61.
[18]Oyamatsu T,Matoyama H,Yamamoto K,et al.A trial for the detection of carp edema virus by using polymerase chain reaction[J]. Aquaculture Science,1997,45(2):247-252.
[19]汪开毓,耿毅,黄锦炉.鱼病诊治彩色图谱[M].北京:中国农业出版社,2011:153-160.
[20]陆承平.兽医微生物学[M].北京:中国农业出版社,2013:351.
[21]Adamek M,Jung-Schroers V,Hellmann J,et al.Concentration of carp edema virus(CEV)DNA in koi tissues affected by koi sleepy disease(KSD)[J].Diseases of Aquatic Organisms,2016,119(3):245-251.
[2] Miyazaki T,Isshiki T,Katsuyuki H.Histopathological and electron microscopy studies on sleepy disease of koi Cyprinus carpio in Japan[J].Diseases of Aquatic Organisms,2005,65(3):197-207.
[3] Murakami Y,Shitanaka M,Toshida S.Studies on mass mortality of juvenile carp:about mass mortality showing edema[J].Bull Hiroshima Fresh Water Fish Exp Stn,1976(1):19-33.
[4] Tomasec I I,Fijan N N.The etiology of infectious dropsy of carp[J].Annals of the New York Academy of Sciences,1965,126(1):606-615.
[5] Oyamatsu T,Hata N,Yamada K,et al.An etiological study on mass mortality of cultured color carp juveniles showing edema[J].Fish Pathology,1997,32(2):81-88.
[6] Way K,Stone D.Emergence of carp edema virus-like(CEV-like)disease in the UK[J].CEFAS Finfish News,2013(15):32-34.
[7] Haenen O,Way K,Stone D,et al.“Koi Sleepy Disease”voor het eerst in Nederland aanqetoond in koikarpers[J].Tijdschr Dierqeneeskd,2014(4):26-29.
[8] Matras M,Borzym E,Stone D,et al.Carp edema virus in Polish aquaculture—evidence of significant sequence divergence and a new lineage in common carp Cyprinus carpio(L.)[J].Journal of Fish Disease,2016,40(3):319-325.
[9] Lewisch E,Gorgoglione B,Way K,et al.Carp edema virus/kio sleepy disease:an emerging disease in central-east Europe[J].Transboundary and Emerging Diseases,2014,62(1):6-12.
[10]Jung-Schroers V,Adamek M,Teitge F,et al.Another potential carp killer carp edema virus disease in Germany[J].BMC Veterinary Research,2015,11(1):114-117.
[11]徐立蒲,王小亮,张文,等.养殖锦鲤鱼浮肿病的检测和鉴定[J].中国畜牧兽医,2017,44(2):613-618.
[12]徐立蒲,王静波,张文,等.鲤鱼浮肿病流行情况的初步研究[J].检验检疫学刊,2016,26(5):14-16.
[13]王小亮,徐立蒲,王姝,等.养殖鲤和锦鲤中的鲤浮肿病毒检测与分析[J].中国畜牧兽医,2017,44(10):213-218.
[14]温智清,刘莹,唐绍林,等.云南锦鲤养殖场鲤浮肿病毒的鉴定及基因型分析[J].病毒学报,2017,33(6):1-9.
[15]Zhang X,Ni Y,Ye J.Carp edema virus,an emerging threat to the carp(Cyprinus carpio)industry in China[J].Aquac Res,2017,474(1):34-39.
[16]吕晓楠,徐立蒲,王姝,等.鲤浮肿病研究进展[J].中国动物检疫,2018,35(5):75-80.
[17]徐立蒲,张文,王姝,等.河南沿黄滩区鲤浮肿病和锦鲤疱疹病毒病初步调查研究[J].科学养鱼,2017(9):59-61.
[18]Oyamatsu T,Matoyama H,Yamamoto K,et al.A trial for the detection of carp edema virus by using polymerase chain reaction[J]. Aquaculture Science,1997,45(2):247-252.
[19]汪开毓,耿毅,黄锦炉.鱼病诊治彩色图谱[M].北京:中国农业出版社,2011:153-160.
[20]陆承平.兽医微生物学[M].北京:中国农业出版社,2013:351.
[21]Adamek M,Jung-Schroers V,Hellmann J,et al.Concentration of carp edema virus(CEV)DNA in koi tissues affected by koi sleepy disease(KSD)[J].Diseases of Aquatic Organisms,2016,119(3):245-251.
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