本文選題：奶牛 + SARA��； 參考：《南京農業(yè)大學》2015年博士論文

【摘要】：瘤胃是反芻動物營養(yǎng)物質消化代謝的主要場所,其健康與否直接關系動物機體健康與生產性能的發(fā)揮。本論文擬系統(tǒng)研究高精料日糧對奶牛瘤胃健康和泌乳性能的影響,研究內容包括以下四方面。1.SARA對瘤胃細菌菌群結構與組成的影響試驗選用4頭裝有瘤胃瘺管的泌乳奶牛,將其隨機分為對照組(CON,40%精料)或SARA組(SARA,70%精料),采用2×2交叉試驗設計。每試驗期第12、17和21天晨飼后0 h采集瘤胃液樣品,提取基因組DNA,進行焦磷酸測序。結果顯示,SARA組奶牛瘤胃pH低于5.8的持續(xù)時間大于5小時,表明SARA模型成功構建。與對照組相比,SARA組奶牛瘤胃乳酸、丙酸、丁酸、戊酸、總VFA和LPS的濃度(P0.05)顯著升高,乙丙比(P0.05)顯著降低。SARA組奶牛瘤胃菌群豐度指數(Chao1和Ace)和多樣性指數(Shannon)顯著低于(P0.05)對照組。在門水平上,SARA組瘤胃變形菌門和擬桿菌門的相對豐度顯著降低(P0.05),但厚壁菌門和放線菌門的相對豐度顯著增加(P0.05)。屬水平上,與對照組相比,SARA組奶牛瘤胃中普雷沃氏菌屬、密螺旋菌屬、厭氧支原體屬、不動桿菌屬、Papillibacter和unclassified Lentisphaerae,and unclassified bacteria的相對豐度顯著降低(P0.05),而瘤胃球菌屬、奇異菌屬、雙歧桿菌和unclassified Clostridiales的相對豐度顯著升高(P0.05)。SARA組奶牛瘤胃內革蘭氏陰性菌的比例顯著下降,且瘤胃LPS濃度與擬桿菌門細菌拷貝數間存在顯著負相關(r =-0.446,P= 0.029)。結果說明,高精料日糧誘發(fā)的SARA可導致瘤胃pH下降,VFA累積,LPS濃度升高;瘤胃細菌菌群結構紊亂,瘤胃細菌菌群的多樣性降低。2.SARA對奶牛瘤胃代謝產物的影響瘤胃微生物菌群的改變與瘤胃代謝密切相關,本章擬通過研究SARA對奶牛代謝產物的影響,進一步解析SARA對瘤胃健康的影響。試驗動物、試驗設計與試驗日糧同上。選用每試驗期第12、17、21天晨飼前瘤胃液樣品,進行GC-MS代謝組學檢測。試驗結果表明,瘤胃液樣品中共檢測到233種化合物,主要包括有機酸類、脂肪酸類、糖類、氨基酸類、嘌呤類、胺類、糖醇類等物質。統(tǒng)計顯示,較對照組相比,SARA組黃嘌呤、次黃嘌呤、尿嘧啶等細菌降解產物含量顯著提高(P0.05);LPS、生物胺、乙醇胺、戊二酸等有毒有害或促炎物質的含量顯著提高(P0.05);丙氨酸、亮氨酸、甘氨酸、谷氨酸、亮氨酸等15種氨基酸的含量顯著提高(P0.05)。對差異代謝物的通路富集分析結果顯示,氨�；鵷RNA生物合成,苯丙氨酸、酪氨酸和色氨酸生物合成,以及纈氨酸、亮氨酸和異亮氨酸生物合成三條通路發(fā)生顯著富集(P0.05)。結果說明,高精料日糧誘發(fā)的SARA會導致瘤胃代謝紊亂,瘤胃內有毒或促炎物質含量升高,氨基酸合成增加,異常代謝產物的增加可能會影響奶牛瘤胃和機體健康。3.SARA對瘤胃上皮基因表達的影響與機制研究本試驗旨在通過基因芯片技術研究SARA對瘤胃上皮基因表達的影響,并結合體外細胞培養(yǎng)手段揭示瘤胃上皮炎癥相關基因對高精料日糧的響應模式及其可能機制。體內試驗:試驗動物、試驗設計與試驗日糧同上。于每期第21 d采集對照組和SARA組瘤胃上皮樣品,進行基因芯片分析。結果顯示,兩組間共有245個基因存在差異。對差異基因進行通路富集分析和功能注釋后,發(fā)現(xiàn)因子受體與其關聯(lián)通路(cytokine-cytokine receptor pathway)顯著富集(P0.05),通路中IL-1β、IL-2、IL-22、CCL19、CCL8、CX3CR1、CXCL6、INHBE、LEPR、PRL和TNFRSF9的表達量在SARA組奶牛瘤胃上皮中顯著上調(P0.05),而通路中IL-6和IL15RA的表達量顯著下調(P0.05),結果表明,瘤胃上皮可能已發(fā)生局部炎癥反應。為驗證基因芯片結果的準確性,使用qRT-PCR對IL-1β、IL-2和IL-6的mRNA表達量進行了測定。相關性分析顯示,瘤胃上皮炎癥因子與瘤胃環(huán)境因子(pH和LPS)存在關聯(lián)(P0.05),為進一步驗證這一結果,進行體外細胞培養(yǎng)試驗。結果說明,瘤胃上皮細胞IL-1β、IL-2、IL-6和IL-8的mRNA表達量在LPS處理后顯著上調(P0.05),而低pH值處理顯著提高了瘤胃上皮細胞TNF-α的表達量,表明瘤胃環(huán)境因子pH和LPS可能在SARA引發(fā)瘤胃局部炎癥反應過程中起著重要作用。4.SARA對奶牛泌乳性能及原奶細菌菌群的影響本章旨在研究SARA對奶牛泌乳性能及乳中細菌菌群結構和組成的影響。試驗動物、試驗設計與試驗日糧同上。分別于每試驗期第17、18和19天采集奶樣,并于每試驗期第12、17和21天飼喂后0 h和4 h采集尾靜脈血,對所采集奶樣中的細菌菌群結構和組成以及所采集血樣中的相關指標進行分析。結果顯示,對照組和SARA組奶牛的干物質采食量(DMI)、產奶量和乳成分無顯著差異(P0.05)。較對照組相比,SARA組奶牛尾靜脈血中白細胞、淋巴細胞數量和白蛋白的含量顯著升高(P0.05),而總蛋白、球蛋白、膽固醇和低密度脂蛋白的含量顯著降低(P0.05)。454測序結果顯示,原奶中的優(yōu)勢菌群為放線菌門、后壁菌門、變形菌門和擬桿菌門。較對照組相比,SARA組奶牛原奶中乳房炎病原菌,包括Stenotrophomonasmaltophilia、Streptococcus parauberis、Brevundimonas diminuta 等細菌的相對豐度顯著提高(P0.05);同時,SARA組奶牛原奶中嗜冷細菌,包括Pseudomonas、Brevundimonas、Sphingobacterium、Alcaligenes、腸桿菌屬和乳桿菌屬等的相對豐度也顯著提高(P0.05)。然而,兩組奶牛原奶中細菌菌群的豐度和多樣性指數(Ace,Chao和Shannon)無顯著差異。結果說明,本試驗中,SARA雖未顯著影響奶牛的泌乳性能,但可能影響奶牛乳腺健康,增加奶�；既榉垦罪L險,并可能降低原奶和乳制品的感官質量,縮短液態(tài)奶的貯存期限。綜上所述,本論文結合組學技術層層漸進、依次揭示了高精料飼喂誘發(fā)的SARA導致瘤胃細菌菌群結構改變,群落趨于簡單化;代謝紊亂,有毒有害或促炎物質含量升高;瘤胃上皮基因表達發(fā)生改變,引發(fā)局部瘤胃炎癥;原奶中細菌菌群中乳房炎致病菌含量增多,增加奶�；既榉垦罪L險。研究結果有助于揭示瘤胃健康與宿主健康的內在關系,為奶牛的健康飼喂提供一定的理論依據。
[Abstract]:The rumen is the main place for nutrient digestion and metabolism of ruminants, and its health is directly related to the health and production performance of the animal body. This paper intends to systematically study the effects of high concentrate diet on the ruminal health and lactating performance of dairy cows. The contents of the study include the following four aspects of the structure and composition of the bacterial flora of the rumen bacteria in the following four aspects. 4 lactating cows with rumen fistula were randomly divided into control group (CON, 40% semen) or group SARA (SARA, 70% semen), and the 2 x 2 cross test was designed. The tumor gastric juice samples were collected at 0 h for 12,17 and 21 days in each test period, and genomic DNA was extracted and sequenced. The results showed that the pH in the rumen of the SARA group was less than 5.8. The duration of the SARA model was more than 5 hours. Compared with the control group, the rumen lactic acid, propionic acid, butyric acid, valerate, valerate, total VFA and LPS concentrations (P0.05) were significantly higher in the SARA group, and the ethylene propylene ratio (P0.05) significantly decreased the abundance index of the rumen group (Chao1 and Ace) and the diversity index (Shannon) of the.SARA group significantly lower than that of the (P0.05) control group At the gate level, the relative abundance of the rumen deformable bacteria gate and the bacteriobacteria in the SARA group decreased significantly (P0.05), but the relative abundance of the actinomycetes and the actinomycetes increased significantly (P0.05). Compared with the control group, the genus Poulet Was, the genus Acinetobacter, Acinetobacter, Papillibacter and UN in the rumen of the group SARA cows. The relative abundance of classified Lentisphaerae and and unclassified bacteria decreased significantly (P0.05), while the relative abundance of rumen, kiwi, Bifidobacterium and unclassified Clostridiales increased significantly (P0.05) in the rumen of the cow's rumen of the cow, and the LPS concentration in the rumen and the bacteriobacteria were copied. There was a significant negative correlation between the number of shellfish (R =-0.446, P= 0.029). The results showed that the SARA induced by high concentrate diet could lead to the decline of the rumen pH, the accumulation of VFA, the increase of LPS concentration, the disorder of the ruminal bacterial flora and the diversity of the rumen bacterial flora, which reduced the effects of.2.SARA on ruminal microbial flora and rumen metabolism. Closely related, this chapter intends to study the effects of SARA on the metabolic products of dairy cows and further analyze the effect of SARA on the health of the rumen. Experimental animals, experimental design and experimental diet are the same. The samples of the tumor and gastric juice before 12,17,21 day morning of each test period were selected for the GC-MS metabolomics test. The results showed that 233 of the samples of the tumor gastric juice detected a total of 233 Compounds, mainly including organic acids, fatty acids, carbohydrates, amino acids, purines, amines, amines, sugar alcohols, etc. statistics show that compared with the control group, the content of the biodegradation products, such as xanthine, hypoxanthine and uracil in the SARA group, is significantly increased (P0.05); LPS, biogenic amines, ethanolamine, glutaric acid and other toxic or probiotic substances The content of 15 amino acids, such as alanine, leucine, glycine, glutamic acid, leucine, and other 15 amino acids increased significantly (P0.05). The analysis of pathway enrichment for differential metabolites showed that amyl tRNA biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, and the biosynthesis of valine, leucine and isoleucine by valine, valine and isoleucine. The pathway was significantly enriched (P0.05). The results showed that the SARA induced by high concentrate diet could lead to the disorder of rumen metabolism, the increase of toxic or proinflammatory substances in the rumen, the increase of amino acid synthesis, and the increase of abnormal metabolites may affect the ruminal gene expression of rumen by the rumen and the body's health.3.SARA. The aim of this study was to study the effect of SARA on the gene expression in the rumen epithelium by gene chip technology, and to reveal the response patterns and possible mechanisms of ruminal dermatitis related genes to high concentrate diet by means of cell culture methods in vitro. In vivo experiments: experimental animals, experimental design and experimental diets, at the same time. A control group and SARA were collected at twenty-first D per period. The results showed that there were 245 differences between the two groups. After the pathway enrichment analysis and functional annotation, we found that the factor receptor and its associated pathway (cytokine-cytokine receptor pathway) were significantly enriched (P0.05), IL-1 beta, IL-2, IL-22, CCL19, CCL8, CX3CR1, CXCL in the pathway. 6, the expression of INHBE, LEPR, PRL and TNFRSF9 was significantly up-regulated in the rumen epithelium of SARA dairy cows (P0.05), while the expression of IL-6 and IL15RA in the pathway was significantly down (P0.05). The results showed that the rumen epithelium may have local inflammatory response. To verify the accuracy of the gene chip results, qRT-PCR is used to express the IL-1 beta, IL-2, and the expressions. The correlation analysis showed that the rumen dermatitis factor was associated with the rumen environmental factors (pH and LPS) (P0.05). In order to further verify this result, the cell culture test was carried out in vitro. The results showed that the mRNA expression of the IL-1 beta, IL-2, IL-6 and IL-8 in the rumen epithelial cells was significantly up-regulated after LPS treatment (P0.05), while the low pH value was treated significantly. The expression of TNF- alpha in the rumen epithelial cells showed that the effects of the rumen environmental factors pH and LPS on the local inflammatory response of the rumen may play an important role in the effects of.4.SARA on lactating performance and the bacterial flora of the dairy cows. This chapter aims to study the effects of SARA on milk performance and the structure and composition of bacterial flora in dairy cows. Experimental animals, experimental design and experimental diet were the same. Milk samples were collected at 17,18 and 19 days of each test period respectively, and 0 h and 4 h were collected at the 12,17 and 21 days of each trial period. The structure and composition of bacterial flora in the milk samples and the related indexes in the collected blood samples were analyzed. The results showed that the control group and the SAR were the SAR. There was no significant difference in the dry matter intake (DMI), milk production and milk composition in the A group (P0.05). Compared with the control group, the white blood cells, the number of lymphocytes and the content of albumin in the tail vein blood of the SARA group increased significantly (P0.05), while the total protein, globulin, cholesterol and low density lipoprotein were significantly reduced (P0.05).454 sequencing results. The dominant bacteria in the original milk were actinomycetes, posterior wall, deformable bacteria and bacteriobacteria. Compared with the control group, the pathogens of mastitis in the milk of SARA dairy cows, including Stenotrophomonasmaltophilia, Streptococcus parauberis, Brevundimonas diminuta and other bacteria, were significantly increased (P0.05), and the original milk of SARA Group dairy cows was also milk. The relative abundance of Pseudomonas, Brevundimonas, Sphingobacterium, Alcaligenes, Enterobacteriaceae and lactobacilli also increased significantly (P0.05). However, there was no significant difference in the abundance and diversity index (Ace, Chao and Shannon) in the two groups of dairy cows. The results showed that in this experiment, SARA did not significantly affect milk. The lactating performance of cattle may affect the health of dairy cows, increase the risk of dairy cow's mastitis, reduce the sensory quality of milk and dairy products and shorten the storage period of liquid milk. In summary, this paper, combined with the technology layer gradually, reveals the structure change of the rumen bacterial flora induced by high concentrate feeding in SARA. The community tends to be simple; metabolic disorders, toxic or harmful or proinflammatory substances increase; the gene expression in the rumen epithelium changes, causes local tumor gastritis; the bacterial flora of the bacterial flora in the original milk increases the risk of mastitis in dairy cows. The results help to reveal the inherent relationship between the health of the rumen and the health of the host, and milk for milk. The health feeding of cattle provides a certain theoretical basis.
【學位授予單位】：南京農業(yè)大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：S858.23

【相似文獻】

相關期刊論文 前10條

1 肖訓軍,孟慶翔;反芻動物瘤胃酸中毒及其調控[J];中國畜牧雜志;2001年05期

2 顧懷君,溫業(yè)嫦;洗胃導胃療法治療牛瘤胃酸中毒[J];貴州畜牧獸醫(yī);2001年04期

3 金朝寧;牛瘤胃酸中毒的糾正[J];青海畜牧獸醫(yī)雜志;2001年02期

4 吳元昌,張銀國,李飛,何高明;一起被誤診的乳牛瘤胃酸中毒[J];畜牧與獸醫(yī);2002年10期

5 吳元昌,張銀國,李飛,何高明;一起被誤診的乳牛瘤胃酸中毒[J];中國奶牛;2002年04期

6 夏兆剛,肖訓軍;反芻動物瘤胃酸中毒發(fā)生機理及防治的研究進展[J];畜禽業(yè);2002年01期

7 趙慶,陳玲,葛廣山,何登峰;一起急性牛瘤胃酸中毒的治療體會[J];中國動物檢疫;2002年04期

8 肖開田;耕牛瘤胃酸中毒的防治[J];貴州畜牧獸醫(yī);2003年02期

9 夏季;羊喂精料過量易發(fā)生瘤胃酸中毒[J];養(yǎng)殖技術顧問;2003年03期

10 李四清;怎樣治療犢牛瘤胃酸中毒[J];河南畜牧獸醫(yī);2003年01期

相關會議論文 前10條

1 張耿;毛勝勇;姚文;朱偉云;;瘤胃酸中毒研究進展[A];動物生理生化學分會第八次學術會議暨全國反芻動物營養(yǎng)生理生化第三次學術研討會論文摘要匯編[C];2004年

2 胡寧璽;薛小蓉;;牛瘤胃酸中毒的治療[A];中國奶業(yè)協(xié)會年會論文集2009（上冊）[C];2009年

3 曹維維;王夢芝;丁洛陽;;反芻動物瘤胃酸中毒的發(fā)生機理及其綜合防治研究進展[A];中國畜牧獸醫(yī)學會2010年學術年會——第二屆中國獸醫(yī)臨床大會論文集(上冊)[C];2010年

4 侯全中;張勝利;;如何防治牛瘤胃酸中毒[A];冀農杯2008“綠色奧運”科技論文集[C];2008年

5 劉燁彤;劉大程;盧德勛;胡紅蓮;;慢性瘤胃酸中毒狀態(tài)下奶山羊瘤胃細菌內幾種相關細菌數量變化的研究[A];中國畜牧獸醫(yī)學會動物營養(yǎng)學分會第十次學術研討會論文集[C];2008年

6 徐俊;;亞急性瘤胃酸中毒診斷方法研究進展[A];第五屆中國畜牧科技論壇論文集[C];2011年

7 羅立平;賀建忠;韓博;;奶牛亞急性瘤胃酸中毒研究新進展[A];中國畜牧獸醫(yī)學會家畜內科學分會第七屆代表大會暨學術研討會論文集（下冊）[C];2011年

8 郭勇慶;曹志軍;李勝利;;奶牛亞急性瘤胃酸中毒診斷及發(fā)病機制研究進展[A];中國畜牧獸醫(yī)學會養(yǎng)牛學分會2011年學術研討會論文集[C];2011年

9 謝正露;張樹坤;姜雪元;葉平生;張源淑;;肝臟對高精料致亞急性瘤胃酸中毒奶山羊的保護效應[A];全國動物生理生化第十二次學術交流會論文摘要匯編[C];2012年

10 楊煥民;柳巨雄;欒新紅;胡仲明;;飼料應激對肉牛HSPs表達的影響[A];中國生理學會第21屆全國代表大會暨學術會議論文摘要匯編[C];2002年

相關重要報紙文章 前10條

1 吉林省大安市 馮國明;羊瘤胃酸中毒的診斷與防治[N];河北農民報;2013年

2 孫志強;冬春養(yǎng)羊要防瘤胃酸中毒[N];河北農民報;2013年

3 內蒙古烏盟農牧學校 張曉東;肉牛瘤胃酸中毒及其預防[N];中國畜牧水產報;2000年

4 李大剛;牛瘤胃酸中毒的防治[N];中國畜牧報;2004年

5 山東省沂水縣三十里獸醫(yī)站 馮培松;肉羊瘤胃酸中毒的防治[N];河北科技報;2010年

6 河北遠征藥業(yè) 王東軍;瘤胃酸中毒[N];中國畜牧獸醫(yī)報;2007年

7 夏道倫;精料喂羊過量 謹防瘤胃酸中毒[N];中國畜牧報;2003年

8 劉國信;羊急性瘤胃酸中毒的防治[N];中國畜牧獸醫(yī)報;2005年

9 河北省畜牧獸醫(yī)研究所 安永福;瘤胃酸中毒的防控[N];河北科技報;2012年

10 河北省畜牧獸醫(yī)研究所 安永福;瘤胃酸中毒的防控[N];河北科技報;2012年

相關博士學位論文 前4條

1 張瑞陽;組學技術研究亞急性瘤胃酸中毒對奶牛瘤胃微生物、代謝和上皮功能的影響[D];南京農業(yè)大學;2015年

2 苑學;肉牛亞急性瘤胃酸中毒微生態(tài)制劑的研制及其效果評價[D];吉林大學;2011年

3 胡紅蓮;奶山羊亞急性瘤胃酸中毒營養(yǎng)生理機制的研究[D];內蒙古農業(yè)大學;2008年

4 李飛;奶山羊亞急性瘤胃酸中毒模型構建與奶牛日糧CBI的優(yōu)化[D];西北農林科技大學;2014年

相關碩士學位論文 前7條

1 路丹;瘤胃異常代謝產物組胺對奶牛肝臟中COX-2表達的影響[D];南京農業(yè)大學;2014年

2 魏德泳;瘤胃酸中毒發(fā)生的微生物學機制及阿卡波糖調控作用的研究[D];南京農業(yè)大學;2010年

3 劉燁彤;慢性酸中毒對瘤胃發(fā)酵功能及瘤胃微生物數量變化的影響[D];內蒙古農業(yè)大學;2009年

4 王婷婷;亞急性瘤胃酸中毒病牛瘤胃組胺對瘤胃上皮細胞炎性通路的影響[D];吉林大學;2015年

5 董淑紅;硫胺素對山羊瘤胃代謝的影響及其與亞急性瘤胃酸中毒關系的研究[D];揚州大學;2011年

6 梅靈;不同產奶水平奶牛瘤胃酸中毒相關指標及血液、乳、糞便中脂多糖濃度的調查研究[D];內蒙古農業(yè)大學;2012年

7 郭立輝;VFA對亞急性瘤胃酸中毒肉牛瘤胃上皮細胞炎性信號通路的影響[D];吉林大學;2015年

，

本文編號：1970565