法氏囊結(jié)構(gòu)發(fā)育與雛雞對IBDV易感性關(guān)系及大蒜合劑抗IBDV作用
發(fā)布時間:2018-03-24 09:41
本文選題:傳染性法氏囊病 切入點(diǎn):大蒜合劑 出處:《中國農(nóng)業(yè)大學(xué)》2016年博士論文
【摘要】:傳染性法氏囊病(Infectious bursal disease, IBD),由傳染性法氏囊病病毒(Infectious bursal disease virus, IBDV)引起的一種雛雞急性、高度接觸性傳染病,主要發(fā)生于3-12周齡的雛雞,其特征性病變?yōu)榉ㄊ夏夷[大、水腫、充血、出血,B淋巴細(xì)胞大量消耗,法氏囊發(fā)育不良,并且會引起嚴(yán)重的體液和細(xì)胞免疫抑制,繼而引起多種疫苗的免疫失敗,增加機(jī)體對其它病原體的易感性,并且耐過雞往往發(fā)育不良,消瘦,貧血生長緩慢,給養(yǎng)禽業(yè)帶來巨大的經(jīng)濟(jì)損失。抗病毒治療僅通過抑制病毒功能但不影響宿主細(xì)胞是比較困難的,因此,尋找既可直接殺死病毒而不傷及機(jī)體組織,又可調(diào)動免疫防御系統(tǒng)的積極性,激發(fā)機(jī)體的主動免疫機(jī)制,增強(qiáng)免疫力,通過主動防御間接發(fā)揮抗病毒作用的藥物意義非凡。大蒜作為食物和藥物已有多年歷史,且長期適量食用不會對機(jī)體造成危害。鑒于此,本研究采用剖檢觀察、臟器指數(shù)測定、H.E.染色、免疫組化染色、透射電子顯微鏡觀察、ELISA、Western blot、流式細(xì)胞術(shù)等技術(shù),通過對傳染性法氏囊病體內(nèi)試驗(yàn)?zāi)P偷膬?yōu)化,大蒜合劑對小鼠的藥效學(xué)試驗(yàn),及中藥大蒜合劑抗傳染性法氏囊病的療效進(jìn)行研究,觀察大蒜合劑是否能夠預(yù)防和解除IBDV所致的免疫抑制,以期為中藥飼料添加劑或預(yù)防劑的合理使用提供可靠的依據(jù)。研究結(jié)果表明:1.傳染性法氏囊病體內(nèi)試驗(yàn)?zāi)P偷暮Y選雛雞對IBDV的易感性與法氏囊的發(fā)育狀況有關(guān),囊小結(jié)髓質(zhì)內(nèi)IgM+B淋巴細(xì)胞數(shù)量多、IgA+B淋巴細(xì)胞數(shù)量少、皮質(zhì)髓質(zhì)分界處的濾泡間上皮及上皮下基膜的完整性破壞是IBDV易感的有利因素。新出雛雛雞囊小結(jié)基膜完整、IgM+B淋巴細(xì)胞數(shù)量少,滴眼感染IBDV不易侵犯髓質(zhì);隨著日齡的增大,IgM+B淋巴細(xì)胞數(shù)量多,皮髓交界處基膜不完整,上皮細(xì)胞層細(xì)胞間出現(xiàn)間隙,雛雞對IBDV變得易感;6周齡后髓質(zhì)中IgM+B淋巴細(xì)胞數(shù)量減少,同時IgA+B淋巴細(xì)胞數(shù)量多,雛雞對IBDV的易感性又變差。且2,3,4,5,6周齡雛雞分別在200 BID50劑量下點(diǎn)眼感染IBDV BC6/85株時,3周齡自來航雞對IBDV的易感性較其余周齡更為敏感,適宜做為中藥抗傳染性法氏囊病的體內(nèi)試驗(yàn)?zāi)P汀?.大蒜合劑的藥效試驗(yàn)口服20 mg/kg以內(nèi)劑量的大蒜合劑能抑制小鼠巨噬細(xì)胞的吞噬能力,降低單核細(xì)胞的活化比率:免疫抑制后,小鼠巨噬細(xì)胞的吞噬能力上升,單核細(xì)胞的活化比率下降,此時口服大蒜合劑能使巨噬細(xì)胞的吞噬能力降低,單核細(xì)胞的活化比率上升。3.大蒜合劑抗傳染性法氏囊病的療效試驗(yàn)飲水口服5 mg/kg大蒜合劑可在一定程度上降低法氏囊內(nèi)病毒載量,對法氏囊淋巴小結(jié)保護(hù)作用較好,提高法氏囊內(nèi)損傷后細(xì)胞重新入駐的比例并有效減輕法氏囊內(nèi)B淋巴細(xì)胞消耗,促進(jìn)法氏囊、胸腺修復(fù);促進(jìn)機(jī)體產(chǎn)生高水平抗體進(jìn)行特異性免疫應(yīng)答反應(yīng);上調(diào)法氏囊內(nèi)IFN-γ的含量進(jìn)行非特異性免疫應(yīng)答反應(yīng);誘導(dǎo)法氏囊內(nèi)淋巴細(xì)胞凋亡,使IBDV靶細(xì)胞減少,抑制細(xì)胞內(nèi)病毒的復(fù)制和增殖。綜上所述,飲水口服5 mg/kg大蒜合劑對IBDV BC6/85毒株引起的機(jī)體損傷具有一定的預(yù)防和治療效果。
[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.
【學(xué)位授予單位】:中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】:博士
【學(xué)位授予年份】:2016
【分類號】:S858.31
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