【摘要】：動物熱應(yīng)激病是一種常發(fā)疾病。國內(nèi)外的研究報道表明,熱應(yīng)激可以對家禽的生長性能、產(chǎn)蛋性能造成嚴(yán)重影響,特別是對機體的免疫功能也會造成嚴(yán)重影響。急性熱應(yīng)激的發(fā)病機理一直是研究熱點。我們參照已報道的急性熱應(yīng)激模型,以肉雞為試驗對象設(shè)置熱應(yīng)激條件。通過臨床癥狀觀察、血液常規(guī)指標(biāo)和生化指標(biāo)檢測、鱟試劑法檢測內(nèi)毒素、組織病理學(xué)及分子生物學(xué)等實驗方法,研究熱應(yīng)激對肉雞免疫功能的影響。主要研究結(jié)果如下:1臨床癥狀及病理學(xué)檢查結(jié)果:隨著熱應(yīng)激時間的持續(xù),肉雞出現(xiàn)精神沉郁、呼吸困難、腹瀉等癥狀。病理學(xué)檢查結(jié)果表明,熱應(yīng)激初期肉雞小腸絨毛、黏膜和黏膜下層未見明顯變化,后期小腸絨毛發(fā)生斷裂、毛細(xì)血管淤血、黏膜上皮細(xì)胞發(fā)生脫落。熱應(yīng)激初期肉雞的脾臟、胸腺和法氏囊出現(xiàn)毛細(xì)血管擴張,充血,到后期免疫器官的淋巴細(xì)胞減少,細(xì)胞核裂解,說明熱應(yīng)激對機體免疫器官損傷隨時間持續(xù)逐漸加劇。2鱟試劑試驗結(jié)果:熱應(yīng)激初期試驗組肉雞血清內(nèi)毒素含量呈極顯著的升高,隨著熱應(yīng)激持續(xù)血清內(nèi)毒素含量明顯升高,熱應(yīng)期后期試驗組內(nèi)毒素含量幾乎是對照組的7倍。在肉雞熱應(yīng)激期間熱應(yīng)激組和VC組內(nèi)毒素含量雖有差異但差異均不顯著。3血常規(guī)試驗結(jié)果:熱應(yīng)激初期肉雞血液紅細(xì)胞數(shù)目輕微升高,到后期顯著降低,并且與對照組相比呈差異顯著;在熱應(yīng)激各時間段,VC組與試驗組紅細(xì)胞數(shù)目相比呈差異不顯著。血液白細(xì)胞在熱應(yīng)激初期顯著降低,而后顯著升高。說明紅細(xì)胞和白細(xì)胞在熱應(yīng)激期間受到明顯的影響,VC對于緩解熱應(yīng)激的影響效果不明顯。4動物生化試驗結(jié)果:試驗組肉雞肝臟功能相關(guān)指標(biāo)ALT、AST持續(xù)升高,與對照組相比,呈差異極顯著,說明肉雞熱應(yīng)激條件下肝臟組織受到損傷,肝臟功能受到影響。腎臟功能相關(guān)指標(biāo)CREA在熱應(yīng)激初期呈顯著升高,隨著熱應(yīng)激時間的持續(xù)升高也越明顯;BUN在熱應(yīng)激初期升高不明顯,隨著熱應(yīng)激時間的持續(xù)BUN呈顯著性升高;TP、GLOB、ALB在熱應(yīng)激初期,試驗組與對照組相比差異顯著,隨著時間持續(xù)差異不顯著。說明肉雞在熱應(yīng)激過程中,肝腎受到損傷的同時,機體的免疫功能也受到抑制。5 RT-PCR法對脾臟內(nèi)毒素受體檢測結(jié)果:與對照組相比,試驗組肉雞脾臟中TLR4受體m RNA的相對表達(dá)量呈先降低后持續(xù)升高的變化趨勢,并且隨著肉雞熱應(yīng)激時間的持續(xù)TLR4受體m RNA的相對表達(dá)量也越明顯,10 h時,熱應(yīng)激組的相對表達(dá)量急速升高約為對照組的1.778倍;而VC組肉雞脾臟中TLR4受體m RNA的相對表達(dá)量呈持續(xù)下降的變化趨勢。這說明大量的內(nèi)毒素進入血液對肉雞免疫功能產(chǎn)生一定的影響。綜上所訴,持續(xù)的熱應(yīng)激能夠引起肝臟細(xì)胞的清除機能下降,肉雞的腸道損傷,大量的腸道內(nèi)毒素進入血液系統(tǒng),形成內(nèi)毒素血癥,隨著血液循環(huán)進入機體的免疫器官,并且隨著熱應(yīng)激時間的持續(xù)對機體的紅細(xì)胞、白細(xì)胞數(shù)目及免疫器官損傷程度加劇;腸源性內(nèi)毒素血癥的發(fā)生導(dǎo)致脾臟內(nèi)毒素受體TLR4 m RNA的表達(dá)上調(diào),說明熱應(yīng)激條件下內(nèi)毒素對肉雞免疫器官具有損傷功能。而VC在緩解熱應(yīng)激所致內(nèi)毒素對肉雞免疫功能損傷方面有一定的作用,但作用不是很明顯。
[Abstract]:Animal heat stress disease is a common disease. Domestic and foreign research reports show that heat stress can seriously affect the growth performance and egg production performance of poultry, especially the immune function of the body. The effects of heat stress on the immune function of broilers were studied by observing clinical symptoms, testing blood routine and biochemical indexes, testing endotoxin by limulus test, histopathology and molecular biology. The results of pathological examination showed that the intestinal villi, mucosa and submucosa of broilers did not change significantly in the early stage of heat stress, but the intestinal villi ruptured, capillaries congested and mucosal epithelial cells shed. Thymus and bursa of Fabricius appeared capillary dilatation, congestion, lymphocyte reduction and nuclear lysis in the immune organs at the late stage, indicating that heat stress on the body's immune organ damage with time continues to intensify. 2 Limulus amebocyte lysate reagent test results: heat stress early experimental group broilers serum endotoxin content showed a very significant increase, with heat stress continued to blood. The content of endotoxin in serum increased significantly, and the content of endotoxin in the late heat stress group was almost 7 times that of the control group. Compared with the control group, there was a significant difference in the number of red blood cells between the VC group and the experimental group at different time points of heat stress, but there was no significant difference in the number of red blood cells between the VC group and the experimental group. The results of biochemical test showed that ALT and AST of broilers in the experimental group increased continuously, which showed that liver tissue was damaged and liver function was affected under heat stress. CREA of kidney function increased significantly in the early stage of heat stress, and increased with the time of heat stress. BUN was not significantly elevated at the initial stage of heat stress, but increased significantly with the duration of heat stress; TP, GLOB, ALB in the initial stage of heat stress, the experimental group was significantly different from the control group, with no significant difference over time. Inhibition.5 RT-PCR assay of spleen endotoxin receptor: Compared with the control group, the relative expression of TLR4 receptor m RNA in the spleen of broiler chickens in the experimental group decreased at first and then continued to increase, and the relative expression of TLR4 receptor m RNA in the broiler chickens increased with the duration of heat stress. The relative expression of TLR4 receptor m RNA in the heat stress group increased at 10 h. The amount of TLR4 receptor m RNA in the spleen of VC broiler chickens showed a decreasing trend, which indicated that a large amount of endotoxin had an effect on the immune function of broiler chickens. Intestinal injury, a large number of intestinal endotoxins into the blood system, forming endotoxemia, with the blood circulation into the body's immune organs, and with the duration of heat stress on the body's red blood cells, white blood cells and immune organ damage increased; intestinal endotoxemia caused the occurrence of splenic endotoxin receptor TLR4 m R The up-regulated expression of NA indicated that endotoxin could damage the immune organs of broilers under heat stress, while VC could alleviate the injury of endotoxin induced by heat stress, but the effect was not obvious.
【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.31

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