抗病毒藥物及甲砜霉素對ALV-J感染致病作用的影響
發(fā)布時間：2018-07-24 12:03
【摘要】：J亞群白血病是由J亞群白血病病毒(Avian Leukosis Virus subgroup J,ALV-J)引起以髓樣細(xì)胞瘤病和血管瘤病為主的傳染性腫瘤性疾病。Panye于1991年首次從肉雞中分離到J亞群白血病病毒,自從該病被報道以來,在許多國家都有病例報道,呈世界性流行。該病在我國早期主要見于各品種的白羽肉雞,后來從肉雞傳播到蛋雞及地方品種雞。該病近年來呈現(xiàn)出宿主范圍擴(kuò)大、發(fā)病日齡提前、腫瘤類型多樣化等新特征�；疾‰u群有較高的死淘率,生產(chǎn)性能下降,給我國養(yǎng)雞業(yè)的健康發(fā)展帶來了巨大威脅。對該病目前尚無有效的疫苗或藥物防制,只能通過種雞群凈化。為尋求我國禽白血病凈化的輔助性藥物,降低凈化成本,推進(jìn)凈化進(jìn)度,本研究擬探究藥物在體內(nèi)外抗ALV-J感染功效及對其致瘤作用的影響。通過CCK-8試劑盒檢測細(xì)胞活力以確定利巴韋林、病毒靈和黃芪多糖對DF1細(xì)胞的最大安全濃度,用ELISA試劑盒檢測ALV-J p27抗原表達(dá)量以確定上述抗病毒藥物體外抗ALV-J活性。結(jié)果表明,利巴韋林、病毒靈和黃芪多糖對DF1細(xì)胞的最大安全濃度分別為0.0391mg/mL、0.0625mg/mL和0.0625 mg/mL。在安全范圍內(nèi),利巴韋林對ALV-J p27的表達(dá)量具有明顯的抑制作用,黃芪多糖也能明顯抑制ALV-J p27的表達(dá),病毒靈對ALV-J p27的表達(dá)基本上沒有抑制作用。為進(jìn)一步驗(yàn)證育雛早期應(yīng)用利巴韋林、病毒靈和黃芪多糖對ALV-J先天性感染雛雞的防治作用,評估育雛早期應(yīng)用對造血及免疫器官有毒副作用的甲砜霉素對ALV-J致瘤作用的影響,通過7日齡SPF雞胚卵黃囊接種1000 TCID50ALV-J,出殼后分組,通過飲水給藥,對各組實(shí)驗(yàn)雞泄殖腔排毒規(guī)律、病毒血癥、疫苗免疫水平、體重、免疫器官指數(shù)、腫瘤發(fā)生率等指標(biāo)進(jìn)行動態(tài)測定,定期剖殺,觀察雞的眼觀及鏡檢病變,評估各種藥物對雛雞先天感染ALV-J致病作用的影響。因條件限制實(shí)驗(yàn)分兩期進(jìn)行,第一期雞胚出殼后隨機(jī)分為3組,兩實(shí)驗(yàn)組雛雞分別定期口服黃芪多糖和甲砜霉素,一組作為病毒感染對照。結(jié)果表明:黃芪多糖和甲砜霉素對ALV-J病毒血癥、ALV-J抗體及疫苗免疫抗體的產(chǎn)生無顯著影響。黃芪多糖組與病毒感染對照組相比體重差異不顯著;甲砜霉素組早期體重明顯高于其他兩組,但后期體重明顯低于另外兩組。甲砜霉素組育雛早期血液白細(xì)胞和淋巴細(xì)胞數(shù)顯著低于另外兩組,60日齡后脾臟和法氏囊的免疫器官指數(shù)顯著降低,與另外兩組差異顯著。腫瘤發(fā)生率以甲砜霉素組最高達(dá)28.57%(10/35),其次是病毒感染對照組20%(7/35),黃芪多糖組最低11.43%(4/35);而且甲砜霉素組腫瘤類型多,包括血管瘤、髓樣細(xì)胞瘤、淋巴肉瘤、纖維肉瘤和腎母細(xì)胞瘤,并且腫瘤病進(jìn)展快,病變重。實(shí)驗(yàn)結(jié)果表明育雛早期應(yīng)用甲砜霉素會導(dǎo)致雛雞免疫抑制,促進(jìn)ALV-J的致瘤作用,并使患病雞腫瘤呈多樣化特征。黃芪多糖對ALV-J的致病性有一定的抑制作用,但未發(fā)現(xiàn)對病毒血癥的影響,兩種藥物對ALV-J致病作用的影響機(jī)制有待于進(jìn)一步研究探討。第二期雞胚出殼后隨機(jī)分為3組,兩實(shí)驗(yàn)組分別定期口服利巴韋林和病毒靈,一組作為病毒感染對照。結(jié)果表明:雖然利巴韋林在DF1細(xì)胞上對ALV-J的增殖具有一定的抑制作用,而且在SPF雞體內(nèi)也能夠提高ND/H9的抗體水平和增加體重,但是對病毒血癥和泄殖腔排毒未見明顯作用;病毒靈在體內(nèi)外均未表現(xiàn)出明顯的抗病毒活性。綜上所述,黃芪多糖可應(yīng)用于ALV-J先天感染雛雞的早期預(yù)防,而育雛早期應(yīng)用甲砜霉素會加重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.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S855.3
本文編號：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.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S855.3
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