法氏囊结构发育与雏鸡对IBDV易感性关系及大蒜合剂抗IBDV作用

发布时间：2018-03-24 09:41

本文选题：传染性法氏囊病　切入点：大蒜合剂　出处：《中国农业大学》2016年博士论文

【摘要】：传染性法氏囊病(Infectious bursal disease, IBD),由传染性法氏囊病病毒(Infectious bursal disease virus, IBDV)引起的一种雏鸡急性、高度接触性传染病,主要发生于3-12周龄的雏鸡,其特征性病变为法氏囊肿大、水肿、充血、出血,B淋巴细胞大量消耗,法氏囊发育不良,并且会引起严重的体液和细胞免疫抑制,继而引起多种疫苗的免疫失败,增加机体对其它病原体的易感性,并且耐过鸡往往发育不良,消瘦,贫血生长缓慢,给养禽业带来巨大的经济损失。抗病毒治疗仅通过抑制病毒功能但不影响宿主细胞是比较困难的,因此,寻找既可直接杀死病毒而不伤及机体组织,又可调动免疫防御系统的积极性,激发机体的主动免疫机制,增强免疫力,通过主动防御间接发挥抗病毒作用的药物意义非凡。大蒜作为食物和药物已有多年历史,且长期适量食用不会对机体造成危害。鉴于此,本研究采用剖检观察、脏器指数测定、H.E.染色、免疫组化染色、透射电子显微镜观察、ELISA、Western blot、流式细胞术等技术,通过对传染性法氏囊病体内试验模型的优化,大蒜合剂对小鼠的药效学试验,及中药大蒜合剂抗传染性法氏囊病的疗效进行研究,观察大蒜合剂是否能够预防和解除IBDV所致的免疫抑制,以期为中药饲料添加剂或预防剂的合理使用提供可靠的依据。研究结果表明：1.传染性法氏囊病体内试验模型的筛选雏鸡对IBDV的易感性与法氏囊的发育状况有关,囊小结髓质内IgM+B淋巴细胞数量多、IgA+B淋巴细胞数量少、皮质髓质分界处的滤泡间上皮及上皮下基膜的完整性破坏是IBDV易感的有利因素。新出雏雏鸡囊小结基膜完整、IgM+B淋巴细胞数量少,滴眼感染IBDV不易侵犯髓质；随着日龄的增大,IgM+B淋巴细胞数量多,皮髓交界处基膜不完整,上皮细胞层细胞间出现间隙,雏鸡对IBDV变得易感；6周龄后髓质中IgM+B淋巴细胞数量减少,同时IgA+B淋巴细胞数量多,雏鸡对IBDV的易感性又变差。且2,3,4,5,6周龄雏鸡分别在200 BID50剂量下点眼感染IBDV BC6/85株时,3周龄自来航鸡对IBDV的易感性较其余周龄更为敏感,适宜做为中药抗传染性法氏囊病的体内试验模型。2.大蒜合剂的药效试验口服20 mg/kg以内剂量的大蒜合剂能抑制小鼠巨噬细胞的吞噬能力,降低单核细胞的活化比率：免疫抑制后,小鼠巨噬细胞的吞噬能力上升,单核细胞的活化比率下降,此时口服大蒜合剂能使巨噬细胞的吞噬能力降低,单核细胞的活化比率上升。3.大蒜合剂抗传染性法氏囊病的疗效试验饮水口服5 mg/kg大蒜合剂可在一定程度上降低法氏囊内病毒载量,对法氏囊淋巴小结保护作用较好,提高法氏囊内损伤后细胞重新入驻的比例并有效减轻法氏囊内B淋巴细胞消耗,促进法氏囊、胸腺修复；促进机体产生高水平抗体进行特异性免疫应答反应；上调法氏囊内IFN-γ的含量进行非特异性免疫应答反应；诱导法氏囊内淋巴细胞凋亡,使IBDV靶细胞减少,抑制细胞内病毒的复制和增殖。综上所述,饮水口服5 mg/kg大蒜合剂对IBDV BC6/85毒株引起的机体损伤具有一定的预防和治疗效果。
[Abstract]:Infectious bursal disease (Infectious bursal, disease, IBD) by infectious bursal disease virus (Infectious bursal disease virus, IBDV) caused a chicken acute, highly contagious disease, mainly in the 3-12 week old chicks, the characteristic lesion for swllen, edema and hyperemia. Hemorrhage, B lymphocyte depletion, bursal dysplasia, and causes severe humoral and cellular immune suppression, then caused the immune failure of multiple vaccine, increased susceptibility to other pathogens, and resistant to chicken dysplasia, emaciation, anemia, slow growth, and bring huge economic losses to poultry industry antiviral. The treatment only by inhibiting the virus but does not affect the function of the host cell is difficult, therefore, looking for can directly kill the virus without harming the body tissue, and can mobilize the enthusiasm of the immune system, stimulate The body of the active immune mechanism, enhance immunity, medicine antiviral effects indirectly through active defense. Special garlic as food and medicine has a long history, and the long-term amount of food will not cause harm to the body. In view of this, this study adopts observation necropsy, organ index determination, H.E. staining, immunohistochemical staining, observation. Transmission electron microscopy ELISA, Western blot, flow cytometry, through optimization of infectious bursal disease in vivo model, effect of garlic mixture on mice experiment, studied the effect of garlic and Chinese medicine mixture against infectious bursal disease, garlic mixture is able to observe the immune prevention and relief by IBDV the suppression, in order to provide a reliable basis for the rational use of traditional Chinese medicine feed additive or preventive agent. The results show that: 1. infectious bursal disease model in vivo test sieve The development status of IBDV and susceptibility of selected chicken bursa, bursa nodule in the medulla of IgM+B lymphocytes, IgA+B lymphocytes, cortex and medulla at the boundaries of the interfollicular epithelium and subepithelial basement membrane integrity damage is the favorable factors in susceptibility to IBDV. The new young chicken bursal nodule intact basement membrane of IgM+B lymphocytes small number of eye drops of IBDV infection is not easy to infringe the medulla; with the age increasing, the number of IgM+B lymphocytes, corticomedullary junction basal membrane, epithelial cell layer between the cell gap, the chicks become susceptible to IBDV; after 6 weeks in the medulla of IgM+B lymphocytes decreased, while the number of IgA+B lymphocytes, chickens susceptibility to IBDV and worse. And 2,3,4,5,6 week old chicks were at the dose of 200 BID50 were infected with IBDV BC6/85 strain, 3 week old chickens. The susceptibility to IBDV is more sensitive than other weeks, Phagocytosis of garlic mixture for oral efficacy test in vivo test model.2. garlic mixture into traditional Chinese medicine anti infectious bursal disease within 20 mg/kg dose can inhibit macrophage activation, reduce the ratio of monocyte immune suppression, increased macrophage phagocytosis, decreased monocyte activation ratio, at this time oral garlic mixture can make macrophage phagocytosis ability decrease, efficacy of oral monocyte activation ratio rose.3. garlic mixture against infectious bursal disease 5 mg/kg garlic mixture can reduce the bursa virus load to a certain extent, good protective effect on cystic lymph nodule Fabricius bursa, improve in the cell injury after re settled and effectively reduce the proportion of bursal B cells in promoting consumption, bursa, thymus and repair; promote the body to produce high levels of resistance into the body Specific immune response; the content of upregulation of bursa of IFN- gamma of non-specific immune response; lymphocyte apoptosis induced by the bursa, the IBDV target cells reduced, inhibition of replication and proliferation of the virus. To sum up, with the preventive and therapeutic effects of body injury drinking 5 mg/kg oral garlic mixture due to the IBDV BC6/85 strain.

【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S858.31

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