中草药饲料添加剂对异育银鲫感染Ⅱ型鲤疱疹病毒的影响
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摘要
分别用添加1%穿心莲、黄芪、板蓝根、大黄4种中草药的定制饲料投喂异育银鲫,而后腹腔注射1mL定量为1.08×10~7个/mL的Ⅱ型鲤疱疹病毒,研究这4种中草药对异育银鲫感染Ⅱ型鲤疱疹病毒的影响。人工感染后采用实时荧光定量PCR的方法测定血液中的Ⅱ型鲤疱疹病毒滴度。试验结果显示,对照攻毒组鱼体临床症状为胸腹部体表大面积充血,鳃部暗红出血,脾脏肿大、内脏出血等体征,为典型的Ⅱ型鲤疱疹病毒发病症状,且用PCR方法能检测出Ⅱ型鲤疱疹病毒基因组片段。投喂含穿心莲、板蓝根、黄芪、大黄的定制饲料,异育银鲫死亡率分别为15%、30%、35%、95%,对照组死亡率为100%。对照组血液中的病毒滴度高于穿心莲、板蓝根、黄芪,大黄与对照组病毒滴度差别较小。研究结果表明,大黄定制饲料抗Ⅱ型鲤疱疹病毒效果相对较差,黄芪与板蓝根抗病毒效果稍强,穿心莲具有较好的抗Ⅱ型鲤疱疹病毒效果,或可作为抗Ⅱ型鲤疱疹病毒的抗病毒饲料产品用于疾病防控。本研究结果为利用中草药药饵防治异育银鲫感染Ⅱ型鲤疱疹病毒提供了一定的理论依据。
Allogyogenetic silver crucian carp Carassius auratus gibelio with body weight of(100±10)g were fed basal diet(as control group),and the basal diet containing 1% of Herba andrographis(group A),Radix astragali(group B),Radix isatidis(group C),and Radix et rhizomarhei(group D)at water temperature of(22±1)℃ for 2 months,and then were injected intraperitoneally with 1 mL(1.08×10~7 ind/mL)of the identified cyprinid herpesvirusⅡto investigate effects of Chinese herbal additives on allogyogenetic silver crucian carp infected cyprinid herpesvirusⅡ.The titer of CyHV-2 virus in the fish blood was measured by real-time fluorescent quantitative PCR after artificial infection.The results showed that the challenged fish were characterized by hyperemia at the base of the fins and on the abdomen,gill bleeding,splenomegaly,and internal bleeding.The mortality of the fish was found to be 15%in group C,30%in group B,35% in group D,and 95% in group A,while there was 100% of mortality in the control group.There was higher virus titer in the control group than those in the treated groups except for group D,indicating that Radix et rhizomarhei showed relatively poor effect against CyHV-2 virus,and that Herba andrographis had better effect against CyHV-2 virus compared with Radix astragali and Radix isatidis,which had potential as a natural feed additive to fight against CyHV-2 virus.The findings provided a theoretical basis for the prevention and treatment of CyHV-2 infection in allogyogenetic silver crucian carp through addition of traditional Chinese herbal medicine.
Allogyogenetic silver crucian carp Carassius auratus gibelio with body weight of(100±10)g were fed basal diet(as control group),and the basal diet containing 1% of Herba andrographis(group A),Radix astragali(group B),Radix isatidis(group C),and Radix et rhizomarhei(group D)at water temperature of(22±1)℃ for 2 months,and then were injected intraperitoneally with 1 mL(1.08×10~7 ind/mL)of the identified cyprinid herpesvirusⅡto investigate effects of Chinese herbal additives on allogyogenetic silver crucian carp infected cyprinid herpesvirusⅡ.The titer of CyHV-2 virus in the fish blood was measured by real-time fluorescent quantitative PCR after artificial infection.The results showed that the challenged fish were characterized by hyperemia at the base of the fins and on the abdomen,gill bleeding,splenomegaly,and internal bleeding.The mortality of the fish was found to be 15%in group C,30%in group B,35% in group D,and 95% in group A,while there was 100% of mortality in the control group.There was higher virus titer in the control group than those in the treated groups except for group D,indicating that Radix et rhizomarhei showed relatively poor effect against CyHV-2 virus,and that Herba andrographis had better effect against CyHV-2 virus compared with Radix astragali and Radix isatidis,which had potential as a natural feed additive to fight against CyHV-2 virus.The findings provided a theoretical basis for the prevention and treatment of CyHV-2 infection in allogyogenetic silver crucian carp through addition of traditional Chinese herbal medicine.
引文
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[10]曹丽萍,丁炜东,张柳,等.香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响[J].水产学报,2008,32(4):628-635.
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[12]李莉娟,罗杨志,刘学芹,等.金鱼鲤疱疹病毒Ⅱ型的分子诊断[J].华中农业大学学报,2013,32(1):92-96.
[13]徐进,曾令兵,杨德国,等.鲤疱疹病毒2型武汉株的分离与鉴定[J].中国水产科学,2013,20(6):1303-1309.
[14]林秀秀,叶元土,吴萍,等.异育银鲫造血器官坏死症病鱼体内鲤疱疹病毒Ⅱ型的电镜观察与超微病理学特征[J].水产学杂志,2016,29(1):17-23.
[15]刘妮,孟以蓉,赵昉.穿心莲水提取物的抗Ⅰ型单纯疱疹病毒作用[J].热带医学杂志,2006(10):1098-1099.
[16]吴莎,金晓晗,施珊珊,等.槲皮素和穿心莲内酯的体外抗病毒作用[J].中药材,2012(12):2003-2006.
[17]艾春汉,邹金虎.6种中药对鲫非特异性免疫效果的影响[J].淡水渔业,2009,39(1):76-79.
[18]刘波,葛鹏彪,周群兰,等.大黄蒽醌提取物对饲养建鲤生长的影响[J].动物学杂志,2007,42(5):141-148.
[19]杨宏亮,黄珂,田珩,等.大黄抑制水产致病菌的研究进展[J].水产科学,2016,35(4):446-452.
[20]Davis J F,Hayasaka S S.The enhancement of resistance of the American eel,Anguilla rostrata Le Sueur,to a pathogenic bacterium,Aeromonas hydrophila,by an extract of the tunicate,Ecteinascidia turbinata[J].Journal of Fish Diseases,2010,7(4):311-316.
[21]洪徐鹏.鲤、鲫疱疹病毒病的诊断与防治措施[J].河北渔业,2013(1):39-40.
[22]俞军,孙琪,王会聪,等.聚六亚甲基胍对异育银鲫造血器官坏死症的防治效果研究[J].淡水渔业,2015,45(4):49-55.
[23]Zhang L,Ma J,Fan Y,et al.Immune response and protection in gibel carp,Carassius gibelio,after vaccination withβ-propiolactone inactivated cyprinid herpesvirus 2[J].Fish&Shellfish Immunology,2016(49):344-350.
[2]Jiang N,Xu J,Ma J,et al.Histopathology and ultrastructural pathology of cyprinid herpesvirusⅡ(CyHV-2)infection in gibel carp,Carassius auratus gibelio[J].Wuhan University Journal of Natural Sciences,2015,20(5):413-420.
[3]Wang L,He J,Liang L,et al.Mass mortality caused by cyprinid herpesvirus 2(CyHV-2)in prussian carp(Carassius gibelio)in China[J].Bulletin-European Association of Fish Pathologists,2012,32(5):164-173.
[4]Xu J,Zeng L,Zhang H,et al.Cyprinid herpesvirus 2infection emerged in cultured gibel carp,Carassius auratus gibelio in China[J].Vet Microb,2013,166(12):138-144.
[5]戈贤平,缪凌鸿,刘波.中草药增强水生动物免疫和抗病能力的研究进展[J].中国渔业质量与标准,2015,5(6):1-2.
[6]王海华.中草药防治水产动物疾病及药理学研究进展[J].中国兽医学杂志,2004(4):37-41.
[7]Harikrishnan R,Balasundaram C,Kim M C,et al.Innate immune response and disease resistance in Carassius auratus by triherbal solvent extracts[J].Fish&Shellfish Immunology,2009,27(3):508-515.
[8]袁春涛,孙方方,李冬梅,等.饲喂两种中药配方对异育银鲫巨噬细胞免疫机能的影响[J].南京大学学报:自然科学版,2007,43(2):152-156.
[9]李俊超,马杰,曾令兵,等.抗锦鲤疱疹病毒的植物药物筛选及其效果比较[J].淡水渔业,2014,44(3):62-67.
[10]曹丽萍,丁炜东,张柳,等.香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响[J].水产学报,2008,32(4):628-635.
[11]Xu L,Podok P,Xie J,et al.Comparative analysis of differential gene expression in kidney tissues of moribund and surviving crucian carp(Carassius auratus gibelio)in response to cyprinid herpesvirus 2infection[J].Archives of Virology,2014,38(8):1961-1974.
[12]李莉娟,罗杨志,刘学芹,等.金鱼鲤疱疹病毒Ⅱ型的分子诊断[J].华中农业大学学报,2013,32(1):92-96.
[13]徐进,曾令兵,杨德国,等.鲤疱疹病毒2型武汉株的分离与鉴定[J].中国水产科学,2013,20(6):1303-1309.
[14]林秀秀,叶元土,吴萍,等.异育银鲫造血器官坏死症病鱼体内鲤疱疹病毒Ⅱ型的电镜观察与超微病理学特征[J].水产学杂志,2016,29(1):17-23.
[15]刘妮,孟以蓉,赵昉.穿心莲水提取物的抗Ⅰ型单纯疱疹病毒作用[J].热带医学杂志,2006(10):1098-1099.
[16]吴莎,金晓晗,施珊珊,等.槲皮素和穿心莲内酯的体外抗病毒作用[J].中药材,2012(12):2003-2006.
[17]艾春汉,邹金虎.6种中药对鲫非特异性免疫效果的影响[J].淡水渔业,2009,39(1):76-79.
[18]刘波,葛鹏彪,周群兰,等.大黄蒽醌提取物对饲养建鲤生长的影响[J].动物学杂志,2007,42(5):141-148.
[19]杨宏亮,黄珂,田珩,等.大黄抑制水产致病菌的研究进展[J].水产科学,2016,35(4):446-452.
[20]Davis J F,Hayasaka S S.The enhancement of resistance of the American eel,Anguilla rostrata Le Sueur,to a pathogenic bacterium,Aeromonas hydrophila,by an extract of the tunicate,Ecteinascidia turbinata[J].Journal of Fish Diseases,2010,7(4):311-316.
[21]洪徐鹏.鲤、鲫疱疹病毒病的诊断与防治措施[J].河北渔业,2013(1):39-40.
[22]俞军,孙琪,王会聪,等.聚六亚甲基胍对异育银鲫造血器官坏死症的防治效果研究[J].淡水渔业,2015,45(4):49-55.
[23]Zhang L,Ma J,Fan Y,et al.Immune response and protection in gibel carp,Carassius gibelio,after vaccination withβ-propiolactone inactivated cyprinid herpesvirus 2[J].Fish&Shellfish Immunology,2016(49):344-350.