【摘要】：敗血癥是指病原菌通過(guò)各種途徑侵入血液,并在血液內(nèi)生長(zhǎng)繁殖產(chǎn)生大量毒素,引起嚴(yán)重的全身性感染。目前敗血癥主要通過(guò)抗生素進(jìn)行治療,但由于抗生素濫用和抗生素殘留帶來(lái)的耐藥性問(wèn)題已越來(lái)越嚴(yán)重。卵黃抗體(Egg Yolk Immunoglobulin, IgY)有高效、安全、不產(chǎn)生耐藥性、無(wú)毒副作用等特點(diǎn)廣泛應(yīng)用于各種疾病的防控。卵黃抗體被視為一種最具應(yīng)用前景的抗生素替代品。但卵黃抗體對(duì)于非腸道疾病的研究較少,本研究建立小鼠敗血癥模型,探討特異性卵黃抗體對(duì)小鼠敗血癥的防治效果。為卵黃抗體對(duì)全身性感染疾病的應(yīng)用研究提供了實(shí)驗(yàn)基礎(chǔ),為卵黃抗體作用機(jī)制的研究提供了新的動(dòng)物模型。 本研究以滅活的金黃色葡萄球菌免疫蛋雞制備特異性卵黃抗體。采用酶聯(lián)免疫吸附法測(cè)定卵黃抗體效價(jià),結(jié)果顯示抗體最高效價(jià)達(dá)到1：18000,抗體高效價(jià)(1：14000)可持續(xù)5周。采用水稀釋-兩次鹽析-超濾-冷凍干燥技術(shù)分離純化卵黃抗體,最終獲得抗體純度達(dá)83.4%。體外抑菌實(shí)驗(yàn)表明,特異性卵黃抗體能有效抑制金黃色葡萄球菌的生長(zhǎng),在卵黃抗體濃度為5mg/ml時(shí)就能起到顯著抑菌效果。免疫熒光結(jié)果表明特異性卵黃抗體能與金黃色葡萄球菌特異性結(jié)合。 用金黃色葡萄球菌通過(guò)尾靜脈注射建立小鼠敗血癥模型,通過(guò)口服或腹腔注射特異性卵黃抗體對(duì)敗血癥小鼠進(jìn)行防治,觀察小鼠健康狀體及死亡率。實(shí)驗(yàn)結(jié)果顯示,通過(guò)尾靜脈注射4×107CFU的金黃色葡萄球菌可致小鼠患敗血癥且死亡率為100%,該劑量為小鼠敗血癥最小致死量。對(duì)小鼠攻毒前采用口服灌胃特異性卵黃抗體進(jìn)行保護(hù),攻毒后持續(xù)3天。小鼠出現(xiàn)寒顫、聚堆、豎毛、行動(dòng)遲緩、眼角有膿狀物,不進(jìn)食、不飲水等不良現(xiàn)象,且與對(duì)照組相比小鼠死亡率無(wú)顯著性變化。而對(duì)小鼠攻毒前通過(guò)腹腔注射特異性卵黃抗體進(jìn)行保護(hù),攻毒后持續(xù)3天,實(shí)驗(yàn)組小鼠死亡率為0%,與對(duì)照組死亡率100%相比防治效果極顯著(P0.01),采用腹腔注射對(duì)小鼠進(jìn)行保護(hù)后小鼠血液中菌數(shù)有顯著降低,小鼠體重減少量與對(duì)照組相比顯著降低(P0.01),實(shí)驗(yàn)結(jié)果說(shuō)明腹腔注射特異性卵黃抗體可對(duì)敗血癥小鼠起到顯著的防治效果,但是口服特異性卵黃抗體對(duì)敗血癥小鼠無(wú)明顯保護(hù)作用。本研究為卵黃抗體應(yīng)用于全身性感染的防治提供了實(shí)驗(yàn)依據(jù)。
[Abstract]:Septicemia refers to pathogenic bacteria invading the blood through various ways, and growing and propagating in the blood to produce a large number of toxins, resulting in serious systemic infection. At present, septicemia is mainly treated by antibiotics, but the problem of drug resistance caused by antibiotic abuse and antibiotic residue has become more and more serious. Yolk antibody (Egg Yolk Immunoglobulin, IgY) has the characteristics of high efficiency, safety, no drug resistance, no toxic and side effects and other characteristics widely used in the prevention and control of various diseases. Yolk antibody is regarded as one of the most promising alternatives to antibiotics. However, there are few studies on yolk antibody for non-intestinal diseases. In this study, a mouse septicemia model was established to explore the preventive and therapeutic effect of specific yolk antibody on septicemia in mice. It provides an experimental basis for the application of yolk antibody to systemic infection diseases and a new animal model for the study of the mechanism of yolk antibody. In this study, inactivated Staphylococcus aureus immunized laying hens were used to prepare specific yolk antibodies. The titer of yolk antibody was determined by enzyme-linked immunosorbent assay (Elisa). The results showed that the highest titer of the antibody was 1 鈮，

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