本文關(guān)鍵詞： 腹瀉 糞腸球菌 腸道微生物 宏基因組學(xué) 16SrDNA　出處：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：腸道微生物與機體的健康息息相關(guān)。仔豬在哺乳期因生理缺陷、外界環(huán)境壓力、食物變化等原因常導(dǎo)致腸道菌群紊亂,發(fā)生腹瀉。腹瀉同樣會加劇腸道微生物的紊亂,生產(chǎn)性能下降,并造成巨大經(jīng)濟損失。有研究表明宿主的腸道微生物與性別相關(guān),但也一些截然相反的結(jié)果。微生態(tài)制劑作為一種理想的抗生素替代品常用來調(diào)節(jié)豬腸道微生物平衡,預(yù)防和治療仔豬腹瀉。本文利用16SrDNA技術(shù)分析腹瀉仔豬腸道菌群結(jié)構(gòu)特征及灌服微生態(tài)制劑對仔豬腸道微生物的內(nèi)在影響及其性別差異。本實驗選擇胎次、產(chǎn)期、產(chǎn)仔數(shù)相近的新生仔豬48窩,觀察仔豬群健康狀況,一旦發(fā)現(xiàn)某窩仔豬發(fā)生腹瀉,立即選擇該窩腹瀉仔豬1頭及同窩同性別未腹瀉的仔豬1頭采集血樣、糞樣,直至采集20窩仔猜樣,并將這20窩仔豬隨機分對照組(C)、試驗組1(T1)、試驗組2(T2)、試驗組3(T3)、試驗組4(T4)、每組4窩。五個組于腹瀉當日(0d)、第3日(3d)、第6日(6d)全窩分別灌服雙蒸水1mL(C)、糞腸球菌2.5 × 108 CFU/mL(T1)、5 × 108 CFU/mL(T2)、1 × 109 CFU/mL(T3)、2 × 109 CFU/mL(T4)的微生態(tài)制劑1mL。到20日齡時,每窩選擇試驗期間發(fā)生過腹瀉與同性別從未腹瀉的仔豬各一頭采集血液樣品和糞樣。試驗分為兩個部分:1腹瀉仔豬腸道微生物、免疫指標的情況本實驗利用16SrDNA技術(shù)分析腹瀉仔豬腸道微生物特征及其性別差異。結(jié)果表明:腹瀉仔豬與未腹瀉仔豬腸道微生物多樣性沒有差異,但在門水平上,腹瀉組疣微菌門Verrucomicrobia(P=0.0419)的豐度顯著高于未腹瀉組;在屬水平上,腹瀉組消化鏈球菌屬Peptostreptococcus(P=0.0195)的豐度顯著高于未腹瀉組,而叢毛單胞菌屬Comamonas(P=0.0308)和Peptoniphilus(=0.0243)的豐度顯著低于未腹瀉組;公仔豬組中 Butyricimonas(P=0.0019)、Sharpea(P=0.0049)和Akkermansia(P=0.0168)的豐度顯著顯著高于母仔豬組。腹瀉仔豬血液中免疫球蛋白M(IgM)濃度顯著低于未腹瀉仔豬組(P=0.049)。2灌服糞腸球菌微生態(tài)制劑對仔豬腸道微生物、免疫指標的影響參考本實驗室前期關(guān)于糞腸球菌微生態(tài)制劑對仔豬腹瀉、血液指標及生長性能影響的研究結(jié)果,挑選對照組(C)、試驗組2(T2)、試驗組4(T4)中同單元、同性別仔豬各6頭,用以研究灌服糞腸球菌微生態(tài)制劑對仔豬腸道微生物的影響。結(jié)果表明,灌服糞腸球菌微生態(tài)制劑有治療腹瀉的功效,但沒有改變仔豬腸道微生物的多樣性和豐度。與對照組相比,灌服微生態(tài)制劑顯著提高血液中IgA(P=0.0250)、IgM(P=0.0402)的濃度。此外,通過兩個時間段對照組腸道微生物的比較,我們發(fā)現(xiàn)仔豬腸道微生物的多樣性和豐度隨日齡改變;在門水平,3-7日齡組梭桿菌門Fusobacteria(P=0.015)的豐度顯著低于20日齡組,但螺旋體門Spirochaetes(P=0.045)和纖維桿菌門Fibrobacteres(P=0.043)的豐度顯著高于20日齡組。
[Abstract]:Intestinal microbes are closely related to the health of the body. During lactation, piglets often suffer from intestinal microflora disorders due to physiological defects, environmental pressures, food changes, and other reasons. Diarrhea also exacerbates intestinal microbiological disorders. Poor productivity and huge economic losses. Studies have shown that intestinal microbes in the host are gender-related. As an ideal substitute for antibiotics, microecological agents are often used to regulate the microbial balance in pig intestines. To prevent and treat diarrhea in piglets, 16s rDNA technique was used to analyze the characteristics of intestinal microflora in piglets, the internal effects of microecological preparations on intestinal microbes and their sex differences. 48 litters of newborn piglets with similar litter size were observed. Once diarrhea was found in one litter piglet, one piglet with diarrhea and one piglet of the same litter with no diarrhea of the same sex were selected to collect blood samples and feces. Until the collection of 20 litter guess, The 20 piglets were randomly divided into two groups: control group (n = 1), test group (n = 1), test group (n = 2), test group (n = 3), test group (n = 3), test group (n = 4), group B (n = 4), each group (n = 4). The five groups were given double distilled water (1 mL C) and enterococcus faecalis (2.5 脳 10 ~ 8 CFU / m L ~ (1) T _ (1)) on the day of diarrhea. A microecological preparation of 1 脳 10 9 CFU / mL T3, 2 脳 10 9 CFU / mL T4). By the age of 20 days, Blood samples and feces were collected from one piglet with diarrhea during each litter selection trial and one piglet with no diarrhea of the same sex. The experiment was divided into two parts: 1. The characteristics of intestinal microbes and their sex differences in diarrhea piglets were analyzed by 16s rDNA technique. The results showed that there was no difference in intestinal microbial diversity between diarrhea piglets and non-diarrhea piglets, but at the portal level, there was no significant difference in intestinal microbial diversity between diarrhea piglets and non-diarrhea piglets. The abundance of Verrucomicrobiae (0.0419) in diarrhea group was significantly higher than that in non-diarrhea group, and the abundance of Peptostreptococcus P0. 0195 in diarrhea group was significantly higher than that in non diarrhea group, while the abundance of Comamonas P0.0308) and Peptoniphilus 0.0243) in diarrhea group was significantly lower than that in non diarrhea group. The abundance of Butyric imonasone P0. 0019 Sharpea P0. 0049) and Akkermania P0. 0168) in male piglets was significantly higher than that in female piglets, and the concentration of immunoglobulin (IgM) in blood of diarrhea piglets was significantly lower than that in non diarrhea piglets (P0. 049.2), and the microecological preparation of Enterococcus faecalis on intestinal microorganism of piglets was significantly lower than that of non diarrhea piglets (P 0. 049. 2). The effect of immune index on the effect of enterococcus faecalis microecological preparation on diarrhea, blood index and growth performance of piglets was studied in our laboratory. The control group was selected as the control group, the control group was divided into two groups: the control group (n = 2), the test group (n = 4) and the control group (n = 6). To study the effect of enterococcus faecalis microecological preparation on intestinal microorganism of piglets. The results showed that enterococcus faecalis microecological preparation had the effect of treating diarrhea. But it did not change the diversity and abundance of intestinal microorganism in piglets. Compared with the control group, the concentration of IgAP0. 0250 in blood was significantly increased by using microecological preparation. In addition, compared with the control group, the intestinal microbes in the control group were compared in two time periods. We found that the diversity and abundance of intestinal microorganism in piglets varied with age, and the abundance of Fusobacteriae (P0. 015) was significantly lower than that of 20 days old group, but the abundance of spirochaetes P0. 045) and Fibrobacteres P0. 043) were significantly higher than those of 20 days old group.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.28

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