發(fā)布時(shí)間：2018-01-30 01:11

  本文關(guān)鍵詞： 雞 益生菌 柔嫩艾美耳球蟲(chóng) 測(cè)序 鼠李糖乳桿菌 盲腸　出處：《河南科技大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：雞的球蟲(chóng)病是由一種或幾種球蟲(chóng)寄生于雞腸道上皮細(xì)胞內(nèi),從而導(dǎo)致腸道組織病變及腸道菌群的破壞。雞球蟲(chóng)病呈全球性分布,在當(dāng)今養(yǎng)禽業(yè)普遍存在且在世界范圍內(nèi)影響較大,集約化養(yǎng)雞業(yè)最為多發(fā),往往造成巨大的經(jīng)濟(jì)損失且對(duì)養(yǎng)禽業(yè)生產(chǎn)發(fā)展產(chǎn)生重大的影響。據(jù)統(tǒng)計(jì),全世界養(yǎng)禽業(yè)每年因雞球蟲(chóng)病導(dǎo)致的經(jīng)濟(jì)損失可達(dá)3億美元。柔嫩艾美耳球蟲(chóng)(Eimeria tenella)俗稱(chēng)盲腸球蟲(chóng),是危害性最為嚴(yán)重的球蟲(chóng)。隨著化學(xué)藥物的不斷使用,禽體內(nèi)藥物殘留和球蟲(chóng)耐藥性的問(wèn)題已經(jīng)成為社會(huì)目前關(guān)注的焦點(diǎn)。因此,尋求一種高效、安全低毒的抗球蟲(chóng)藥物已然成為當(dāng)前亟需解決的問(wèn)題。本研究通過(guò)對(duì)盲腸微生物測(cè)序發(fā)現(xiàn),以益生菌枯草芽孢桿菌、鼠李糖乳桿菌和酵母菌為添加物來(lái)探討益生菌對(duì)E.tenella感染雞的保護(hù)效果研究。主要通過(guò)下面幾個(gè)實(shí)驗(yàn)進(jìn)行研究:(1)E.tenella感染雞盲腸微生物的測(cè)序建立E.tenella感染模型,分為對(duì)照組和感染組,在感染第14天后,分別采集感染組和對(duì)照組盲腸組織,提取盲腸內(nèi)容物微生物基因組DNA,然后對(duì)其進(jìn)行測(cè)序。實(shí)驗(yàn)結(jié)果顯示:在Alpha多樣性分析中,感染組與對(duì)照組相比,感染組ACE的值增加16.86,Chao1值增加18.4,Simpson值增加0.01;E.tenella感染后對(duì)微生物群落影響較大,改變了雞盲腸內(nèi)微生物不同菌群在門(mén)和屬水平上的相對(duì)豐度;在Beta多樣性分析中,對(duì)照組的樣品間空間距離較近而感染組的樣品間空間距離較遠(yuǎn)。(2)益生菌對(duì)E.tenella感染雞保護(hù)作用建立E.tenella感染雞試驗(yàn)?zāi)Ｐ?以臨床癥狀、血便記分、存活率、增重率、和益生菌對(duì)E.tenella的保護(hù)效果研究。結(jié)果顯示:益生菌的添加能減輕球蟲(chóng)感染雞的血便計(jì)分,顯著降低了球蟲(chóng)感染雞的死亡率,提高球蟲(chóng)病感染雞的增重率。益生菌抗球蟲(chóng)指數(shù)為137,為低效抗球蟲(chóng)試劑。表明益生菌可以與其它飼料添加劑聯(lián)合應(yīng)用于臨床球蟲(chóng)病的防治。(3)益生菌對(duì)E.tenella感染雞盲腸組織結(jié)構(gòu)及免疫細(xì)胞的影響建立E.tenella感染雞試驗(yàn)?zāi)Ｐ?在接種球蟲(chóng)第7天,稱(chēng)重后取雞盲腸組織,用HE染色觀察盲腸組織病理學(xué)變化,采用AB-PAS和PAS染色法,來(lái)觀察E.tenella感染雞盲腸杯狀細(xì)胞糖原和粘液成分的變化。實(shí)驗(yàn)結(jié)果表明:對(duì)照組,盲腸黏膜層的糖原及粘液成份分布正常,排列有序。E.tenella感染后,造成盲腸黏膜層被破壞,大量的糖原及粘液丟失。結(jié)論:E.tenella感染雞后改變了雞盲腸內(nèi)微生物不同菌群在門(mén)和屬水平上的相對(duì)豐度;在E.tenella感染后添加益生菌能有效減輕雞的糞便計(jì)分,提高球蟲(chóng)病感染雞的增重率,提高了雞的抗球蟲(chóng)指數(shù)和感染雞的存活率;益生菌呈現(xiàn)出低效抗球蟲(chóng)的作用;同時(shí),添加益生菌可減輕盲腸腸黏膜和腸腺組織的病理學(xué)結(jié)構(gòu)的損傷,明顯改善雞的腸道粘膜免疫,進(jìn)而發(fā)揮了抗球蟲(chóng)作用。
[Abstract]:Coccidiosis of chicken is parasitic by one or more coccidiosis in the intestinal epithelial cells, resulting in intestinal tissue lesions and the destruction of intestinal flora. Chicken coccidiosis is distributed globally. In today's poultry industry is widespread and has a greater impact in the world, intensive chicken industry is the most frequent, often resulting in huge economic losses and the development of poultry industry has a significant impact. According to statistics. The annual economic loss caused by chicken coccidiosis in the world poultry industry can reach US $300 million. Eimteneria tenella (Eimteneria ella) is commonly known as cecum coccidia. With the continuous use of chemical drugs, the problems of drug residues and coccidiosis resistance in poultry have become the focus of attention. Therefore, to seek a high efficiency. Safety and low toxicity anti-coccidiosis drugs have become a problem that needs to be solved. Through sequencing of cecum microbes, we found the probiotics Bacillus subtilis. The protective effect of probiotics on E. tenella infected chicken was studied by using Lactobacillus rhamnoides and yeast as additives. E. tenella infection model was established by sequencing of E. tenella infected chicken cecum microbes. It was divided into control group and infection group. After 14 days of infection, the cecum tissues of infection group and control group were collected, and the microbial genomic DNA of cecum contents were extracted. Then it was sequenced. The results showed that in the analysis of Alpha diversity, the value of ACE in the infected group was increased by 16.86%, and the value of Chao1 was increased by 18.40% in the infected group as compared with the control group. Simpson increased by 0.01; E. tenella infection had a great influence on the microbial community, which changed the relative abundance of different microflora in chicken cecum at phylum and genus level. In Beta diversity analysis. The experimental model of E. tenella infected chicken was established by probiotics on the protection of E. tenella infected chicken. The clinical symptoms, blood stool score, survival rate, weight gain rate and the protective effect of probiotics on E. tenella were studied. The results showed that probiotics could reduce the blood stool score of coccidiosis infected chicken. The mortality rate of chickens infected with coccidiosis was significantly reduced and the weight gain rate of chickens infected with coccidiosis was increased. The probiotic index against coccidiosis was 137. It is indicated that probiotics can be combined with other feed additives to control clinical coccidiosis. Effects of probiotics on the tissue structure and immune cells of the cecum of E. tenella infected chicken, the experimental model of E. tenella infected chicken was established. On the 7th day of inoculation of coccidia, the cecum tissues were taken from chickens after weighing, and the histopathological changes of cecum were observed by HE staining. AB-PAS and PAS staining were used. To observe the changes of glycogen and mucus in goblet cells of the caecum infected with E. tenella. The results showed that the glycogen and mucus in the mucosal layer of the caecum were normal in the control group. After infection, the cecum mucous layer was destroyed. Conclusion the relative abundance of different microflora in the cecum of chicken was changed after infection with E. tenella at the phylum and genus level. The addition of probiotics after E. tenella infection can effectively reduce the fecal score of chickens, increase the weight gain rate of coccidiosis infected chickens, improve the anti-coccidiosis index and the survival rate of infected chickens. Probiotics showed low effect against coccidiosis. At the same time, the addition of probiotics could reduce the pathological structure of caecum mucosa and intestinal gland, improve the intestinal mucosal immunity of chicken, and then play an anti-coccidial effect.
【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S858.31

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