抗病毒药物及甲砜霉素对ALV-J感染致病作用的影响
发布时间：2018-07-24 12:03
【摘要】：J亚群白血病是由J亚群白血病病毒(Avian Leukosis Virus subgroup J,ALV-J)引起以髓样细胞瘤病和血管瘤病为主的传染性肿瘤性疾病。Panye于1991年首次从肉鸡中分离到J亚群白血病病毒,自从该病被报道以来,在许多国家都有病例报道,呈世界性流行。该病在我国早期主要见于各品种的白羽肉鸡,后来从肉鸡传播到蛋鸡及地方品种鸡。该病近年来呈现出宿主范围扩大、发病日龄提前、肿瘤类型多样化等新特征。患病鸡群有较高的死淘率,生产性能下降,给我国养鸡业的健康发展带来了巨大威胁。对该病目前尚无有效的疫苗或药物防制,只能通过种鸡群净化。为寻求我国禽白血病净化的辅助性药物,降低净化成本,推进净化进度,本研究拟探究药物在体内外抗ALV-J感染功效及对其致瘤作用的影响。通过CCK-8试剂盒检测细胞活力以确定利巴韦林、病毒灵和黄芪多糖对DF1细胞的最大安全浓度,用ELISA试剂盒检测ALV-J p27抗原表达量以确定上述抗病毒药物体外抗ALV-J活性。结果表明,利巴韦林、病毒灵和黄芪多糖对DF1细胞的最大安全浓度分别为0.0391mg/mL、0.0625mg/mL和0.0625 mg/mL。在安全范围内,利巴韦林对ALV-J p27的表达量具有明显的抑制作用,黄芪多糖也能明显抑制ALV-J p27的表达,病毒灵对ALV-J p27的表达基本上没有抑制作用。为进一步验证育雏早期应用利巴韦林、病毒灵和黄芪多糖对ALV-J先天性感染雏鸡的防治作用,评估育雏早期应用对造血及免疫器官有毒副作用的甲砜霉素对ALV-J致瘤作用的影响,通过7日龄SPF鸡胚卵黄囊接种1000 TCID50ALV-J,出壳后分组,通过饮水给药,对各组实验鸡泄殖腔排毒规律、病毒血症、疫苗免疫水平、体重、免疫器官指数、肿瘤发生率等指标进行动态测定,定期剖杀,观察鸡的眼观及镜检病变,评估各种药物对雏鸡先天感染ALV-J致病作用的影响。因条件限制实验分两期进行,第一期鸡胚出壳后随机分为3组,两实验组雏鸡分别定期口服黄芪多糖和甲砜霉素,一组作为病毒感染对照。结果表明:黄芪多糖和甲砜霉素对ALV-J病毒血症、ALV-J抗体及疫苗免疫抗体的产生无显著影响。黄芪多糖组与病毒感染对照组相比体重差异不显著;甲砜霉素组早期体重明显高于其他两组,但后期体重明显低于另外两组。甲砜霉素组育雏早期血液白细胞和淋巴细胞数显著低于另外两组,60日龄后脾脏和法氏囊的免疫器官指数显著降低,与另外两组差异显著。肿瘤发生率以甲砜霉素组最高达28.57%(10/35),其次是病毒感染对照组20%(7/35),黄芪多糖组最低11.43%(4/35);而且甲砜霉素组肿瘤类型多,包括血管瘤、髓样细胞瘤、淋巴肉瘤、纤维肉瘤和肾母细胞瘤,并且肿瘤病进展快,病变重。实验结果表明育雏早期应用甲砜霉素会导致雏鸡免疫抑制,促进ALV-J的致瘤作用,并使患病鸡肿瘤呈多样化特征。黄芪多糖对ALV-J的致病性有一定的抑制作用,但未发现对病毒血症的影响,两种药物对ALV-J致病作用的影响机制有待于进一步研究探讨。第二期鸡胚出壳后随机分为3组,两实验组分别定期口服利巴韦林和病毒灵,一组作为病毒感染对照。结果表明:虽然利巴韦林在DF1细胞上对ALV-J的增殖具有一定的抑制作用,而且在SPF鸡体内也能够提高ND/H9的抗体水平和增加体重,但是对病毒血症和泄殖腔排毒未见明显作用;病毒灵在体内外均未表现出明显的抗病毒活性。综上所述,黄芪多糖可应用于ALV-J先天感染雏鸡的早期预防,而育雏早期应用甲砜霉素会加重ALV-J的致瘤作用。
[Abstract]:J subgroup leukemia is a J subgroup leukemia virus (Avian Leukosis Virus subgroup J, ALV-J) caused by myeloid cytomonosis and hemangioma disease, which is the first infectious tumor disease,.Panye was first isolated from broilers in 1991 to the J subgroup leukemia virus. Since it has been reported, there have been cases reported in many countries and are worldwide. In the early stage of China, the disease was mainly found in various varieties of white feather broilers, then spread from broilers to laying hens and local breeds. In recent years, the disease has shown a wide range of host range, early onset of onset, and a variety of tumor types. There is a great threat to the disease. There is no effective vaccine or drug control for the disease. It can only be purified by the fowl group. In order to seek the auxiliary drugs for the purification of avian leukosis in our country, reduce the cost of purification and advance the progress of purification, this study intends to explore the effect of drugs on the anti ALV-J infection in vivo and in vivo and its effect on its tumorigenesis. Through the examination of the CCK-8 Kit Cell viability was measured to determine the maximum safety concentration of Leigh Bhave Lin, virus and Astragalus Polysaccharides on DF1 cells. The expression of ALV-J p27 antigen was detected by ELISA kit to determine the anti ALV-J activity of the above antiviral drugs. The results showed that the maximum safety concentration of Leigh Bhave Lin, virus and astragalus polysaccharides to DF1 cells was 0.0391mg/mL, respectively, 0. 0625mg/mL and 0.0625 mg/mL. were in a safe range, Leigh Bhave Lin had obvious inhibitory effect on the expression of ALV-J p27, and astragalus polysaccharides could obviously inhibit the expression of ALV-J p27. Virus spirit had no inhibitory effect on the expression of ALV-J p27. In order to further verify the early stage of brooding, we should use Leigh Bhave Lin, virus and astragalus polysaccharides to ALV-J first. The effect of the natural infection of chicks on the effect of the early application of the toxic side effects of the brood to the hemopoietic and immune organs of the ALV-J, through the inoculation of 1000 TCID50ALV-J through the yolk sac of the 7 day old SPF chicken embryo, the group after the shell and the drinking water, and the test of the law of detoxification of the chicken cloaca, the viremia and the immunization of the vaccine The indexes of level, weight, immune organ index, tumor incidence and so on were dynamically measured, regular caesarean section, observation of the eye view and pathological changes of chickens, and the effects of various drugs on the pathogenesis of ALV-J in chick congenital infection. The first stage chicken embryos were divided into 3 groups randomly. The first stage chicken embryos were randomly divided into 3 groups, and the two experimental chicks were given regular mouth. The results showed that Astragalus Polysaccharides and methyl sulfonamycin had no significant influence on ALV-J virus, ALV-J antibody and immunization antibody. The weight difference between astragalus polysaccharide group and virus infection control group was not significant, and the early body weight of methylsulfonamicin group was significantly higher than that of the other two groups. But the post body weight was significantly lower than that of the other two groups. The number of leukocytes and lymphocytes in the early stage of brood was significantly lower than the other two groups. The immune organ index of the spleen and the bursa of the Fabricius decreased significantly after 60 days of age, and the difference was significant between the other two groups. The incidence of the tumor was 28.57% (10/35) in the formamycin group, followed by the virus infection control. Group 20% (7/35), the lowest 11.43% (4/35) in the astragalus polysaccharide group, and many types of tumorigenin group, including hemangioma, myeloid cytoma, lymphosarcoma, fibrosarcoma and nephroblastoma, and the progression of the tumor is fast, and the pathological changes are heavy. The experimental results show that the early use of sulfoxamin can lead to the immunosuppression of chicks and promote the tumorigenicity of ALV-J. The disease of the sick chicken showed a variety of characteristics. Astragalus polysaccharide had a certain inhibitory effect on the pathogenicity of ALV-J, but it had not found the effect on the viremia. The mechanism of the effect of the two drugs on the pathogenesis of ALV-J needs to be further studied. The second stage chicken embryos were randomly divided into 3 groups, and the two experimental groups were regularly orally orally taken to Leigh Bhave Lin and The results showed that although Leigh Bhave Lin had a certain inhibitory effect on the proliferation of ALV-J in DF1 cells, and in the SPF chicken, it could increase the level of antibody and increase the body weight of ND/H9, but it had no obvious effect on viremia and the detoxification of the cloaca. In summary, astragalus polysaccharides can be used in the early prevention of ALV-J infective chicks, and the early use of sulfoxamiin can aggravate the ALV-J's tumorigenicity.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S855.3
本文编号：2141319
[Abstract]:J subgroup leukemia is a J subgroup leukemia virus (Avian Leukosis Virus subgroup J, ALV-J) caused by myeloid cytomonosis and hemangioma disease, which is the first infectious tumor disease,.Panye was first isolated from broilers in 1991 to the J subgroup leukemia virus. Since it has been reported, there have been cases reported in many countries and are worldwide. In the early stage of China, the disease was mainly found in various varieties of white feather broilers, then spread from broilers to laying hens and local breeds. In recent years, the disease has shown a wide range of host range, early onset of onset, and a variety of tumor types. There is a great threat to the disease. There is no effective vaccine or drug control for the disease. It can only be purified by the fowl group. In order to seek the auxiliary drugs for the purification of avian leukosis in our country, reduce the cost of purification and advance the progress of purification, this study intends to explore the effect of drugs on the anti ALV-J infection in vivo and in vivo and its effect on its tumorigenesis. Through the examination of the CCK-8 Kit Cell viability was measured to determine the maximum safety concentration of Leigh Bhave Lin, virus and Astragalus Polysaccharides on DF1 cells. The expression of ALV-J p27 antigen was detected by ELISA kit to determine the anti ALV-J activity of the above antiviral drugs. The results showed that the maximum safety concentration of Leigh Bhave Lin, virus and astragalus polysaccharides to DF1 cells was 0.0391mg/mL, respectively, 0. 0625mg/mL and 0.0625 mg/mL. were in a safe range, Leigh Bhave Lin had obvious inhibitory effect on the expression of ALV-J p27, and astragalus polysaccharides could obviously inhibit the expression of ALV-J p27. Virus spirit had no inhibitory effect on the expression of ALV-J p27. In order to further verify the early stage of brooding, we should use Leigh Bhave Lin, virus and astragalus polysaccharides to ALV-J first. The effect of the natural infection of chicks on the effect of the early application of the toxic side effects of the brood to the hemopoietic and immune organs of the ALV-J, through the inoculation of 1000 TCID50ALV-J through the yolk sac of the 7 day old SPF chicken embryo, the group after the shell and the drinking water, and the test of the law of detoxification of the chicken cloaca, the viremia and the immunization of the vaccine The indexes of level, weight, immune organ index, tumor incidence and so on were dynamically measured, regular caesarean section, observation of the eye view and pathological changes of chickens, and the effects of various drugs on the pathogenesis of ALV-J in chick congenital infection. The first stage chicken embryos were divided into 3 groups randomly. The first stage chicken embryos were randomly divided into 3 groups, and the two experimental chicks were given regular mouth. The results showed that Astragalus Polysaccharides and methyl sulfonamycin had no significant influence on ALV-J virus, ALV-J antibody and immunization antibody. The weight difference between astragalus polysaccharide group and virus infection control group was not significant, and the early body weight of methylsulfonamicin group was significantly higher than that of the other two groups. But the post body weight was significantly lower than that of the other two groups. The number of leukocytes and lymphocytes in the early stage of brood was significantly lower than the other two groups. The immune organ index of the spleen and the bursa of the Fabricius decreased significantly after 60 days of age, and the difference was significant between the other two groups. The incidence of the tumor was 28.57% (10/35) in the formamycin group, followed by the virus infection control. Group 20% (7/35), the lowest 11.43% (4/35) in the astragalus polysaccharide group, and many types of tumorigenin group, including hemangioma, myeloid cytoma, lymphosarcoma, fibrosarcoma and nephroblastoma, and the progression of the tumor is fast, and the pathological changes are heavy. The experimental results show that the early use of sulfoxamin can lead to the immunosuppression of chicks and promote the tumorigenicity of ALV-J. The disease of the sick chicken showed a variety of characteristics. Astragalus polysaccharide had a certain inhibitory effect on the pathogenicity of ALV-J, but it had not found the effect on the viremia. The mechanism of the effect of the two drugs on the pathogenesis of ALV-J needs to be further studied. The second stage chicken embryos were randomly divided into 3 groups, and the two experimental groups were regularly orally orally taken to Leigh Bhave Lin and The results showed that although Leigh Bhave Lin had a certain inhibitory effect on the proliferation of ALV-J in DF1 cells, and in the SPF chicken, it could increase the level of antibody and increase the body weight of ND/H9, but it had no obvious effect on viremia and the detoxification of the cloaca. In summary, astragalus polysaccharides can be used in the early prevention of ALV-J infective chicks, and the early use of sulfoxamiin can aggravate the ALV-J's tumorigenicity.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S855.3
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