奶牛乳房炎與腸道菌群的關(guān)聯(lián)研究
發(fā)布時間：2019-04-29 07:31
【摘要】：乳房炎是奶牛養(yǎng)殖場中的高發(fā)病,經(jīng)抗生素治愈后極易復(fù)發(fā)。疾病的反復(fù)發(fā)作導(dǎo)致奶牛的產(chǎn)奶量持續(xù)下降,乳頭壞死,進而被淘汰。乳房炎也是哺乳期婦女常患的疾病,傳統(tǒng)的抗生素治療損傷新生兒的神經(jīng)系統(tǒng)。在乳房炎的治療方法中,抗生素因其大量的負面作用使得其已不適合在乳房炎治療中濫用,應(yīng)加快開發(fā)新的且有效的抗生素替代物用以治療或預(yù)防乳房炎。益生菌在多種疾病的預(yù)防與治療中都有成功的應(yīng)用實例。因此,益生菌在乳房炎的治療與預(yù)防中是否真的有效是一個有趣的研究方向。益生菌主要于宿主的腸道中發(fā)揮其關(guān)鍵作用,因此腸道菌群與乳房炎發(fā)病是否有關(guān)聯(lián),是益生菌能否應(yīng)用于乳房炎的治療中需要解答的首個問題。在本論文中,以健康奶牛與患乳房炎奶牛為對象,對健康奶牛與乳房炎奶牛的腸道菌群中常見的菌屬與菌種進行定量研究,相關(guān)性分析結(jié)果發(fā)現(xiàn)在兩組奶牛之間,以L.buchneri為首的乳桿菌種對兩組差異的貢獻度最大,達到27.32%,其次為L.sake,達到9.64%。選定哺乳期無菌小鼠為動物模型,將乳房炎奶牛糞便與健康奶牛糞便分別移植入無菌小鼠體內(nèi),同時選取一組移植乳房炎奶牛糞便的小鼠進行益生菌干預(yù)。通過菌群移植,對哺乳期小鼠的乳腺組織以及身體其它各項免疫指標進行監(jiān)控,探究移植健康奶牛糞便小鼠組與移植乳房炎奶牛糞便小鼠組以及移植乳房炎奶牛糞便同時進行益生菌干預(yù)組小鼠的乳腺組織是否發(fā)生炎癥以及炎癥程度如何。實驗結(jié)果發(fā)現(xiàn),移植乳房炎奶牛糞便的小鼠在菌群移植25天檢測時發(fā)現(xiàn)明顯的乳腺組織炎癥反應(yīng),而移植健康奶牛糞便的小鼠沒有明顯的炎癥,進行益生菌干預(yù)的小鼠組炎癥較弱。在組織病理學(xué)檢驗之后,采集三組小鼠糞便樣品與兩組奶牛糞便樣品一同進行了宏基因組測序以及16S rRNA基因全長測序。結(jié)果發(fā)現(xiàn),奶牛腸道菌群移植入小鼠體內(nèi)之后,其結(jié)構(gòu)發(fā)生改變,但是在移植健康組奶牛糞便的小鼠組中,腸道菌群的結(jié)構(gòu)改變較小,更傾向于健康組奶牛的腸道菌群結(jié)構(gòu)；移植乳房炎奶牛糞便的兩組小鼠,其中一組進行益生菌干預(yù)的小鼠組,兩組小鼠的腸道菌群結(jié)構(gòu)區(qū)分度較小,但與乳房炎奶牛的腸道菌群結(jié)構(gòu)改變較大。同時在健康組奶牛與移植其糞便的小鼠組腸道菌群中產(chǎn)乙酸、丁酸以及琥珀酸的菌屬豐度較高。16S rRNA基因全長測序發(fā)現(xiàn)在移植健康奶牛糞便的小鼠中Lactobacillus的含量較高。從宏基因組測序的結(jié)果與16S rRNA基因全長測序結(jié)果發(fā)現(xiàn),患乳房炎奶牛與健康奶牛的腸道菌群結(jié)構(gòu)與功能都存在差異,且將奶牛腸道菌群移植入無菌小鼠之后,這種菌群結(jié)構(gòu)與功能之上的差異同時被放大形成鮮明的對比。實驗結(jié)果表明健康的腸道菌群結(jié)構(gòu)對于保持奶牛的健康狀態(tài)有直接的影響,益生菌可調(diào)節(jié)奶牛的腸道菌群,對于維持奶牛健康,預(yù)防和緩解乳房炎有不可忽視的潛能。
[Abstract]:Mastitis is a high incidence in dairy cattle farms and easy to recur after being cured by antibiotics. The recurrence of the disease led to a continuous decline in milk production, nipple necrosis, and thus elimination of milk production in cows. Mastitis is also a common disease among lactating women. Traditional antibiotics treat neonatal nervous system damage. In the treatment of mastitis, because of its negative effects, antibiotics are no longer suitable for abuse in mastitis treatment. It is necessary to speed up the development of new and effective antibiotic substitutes for the treatment or prevention of mastitis. Probiotics have been successfully applied in the prevention and treatment of many diseases. Therefore, whether probiotics is really effective in the treatment and prevention of mastitis is an interesting research direction. Probiotics play a key role in the host intestine, so whether the intestinal flora is associated with mastitis is the first question whether probiotics can be used in the treatment of mastitis. In this paper, healthy cows and cows with mastitis were used to quantitatively study the common bacteria and bacteria in the intestinal microflora of healthy cows and mastitis cows. The results of correlation analysis showed that between the two groups of cows, the results of correlation analysis showed that there was no significant difference between the two groups of cows. The difference between the two groups was the largest by L.buchneri, with 27.32%, followed by L.sake, which reached 9.64%. Lactating aseptic mice were selected as animal models. Mastitis cow feces and healthy cow feces were transplanted into aseptic mice, and a group of mice transplanted mastitis cow feces were selected for probiotics intervention. Through microflora transplantation, breast tissue and other immune indexes of lactating mice were monitored. To investigate whether the mammary tissue of the mice transplanted with healthy cow feces and the mice transplanted with mastitis cow feces and the mice treated with probiotics at the same time had inflammation and the degree of inflammation. The results showed that mice transplanted with mastitis cow feces showed significant inflammation in breast tissue during 25 days of microflora transplantation, but no significant inflammation was found in mice transplanted with cow feces from healthy cows. The inflammation of mice treated with probiotics was weaker than that of mice treated with probiotics. After histopathological examination, three groups of mice fecal samples and two groups of cow fecal samples were collected and sequenced for metagenomic sequencing and 16s rRNA gene full-length sequencing. The results showed that the structure of intestinal flora was changed after it was transplanted into mice, but in the group of mice transplanted with cow feces, the structure change of intestinal flora was smaller, and it was more inclined to the structure of intestinal flora of cow in healthy group. Two groups of mice transplanted mastitis cow feces, one group of mice treated with probiotics, the intestinal microflora structure of the two groups of mice was smaller, but with mastitis cow intestinal flora structure change is greater. At the same time, the abundance of bacteria producing acetic acid, butyric acid and succinic acid was higher in the intestinal flora of healthy cows and mice transplanted with their feces. The full-length sequencing of 16s rRNA gene showed that the content of Lactobacillus in the feces of transplanted healthy cows was higher. From the results of macro-genome sequencing and full-length 16s rRNA gene sequencing, it was found that there were differences in the structure and function of intestinal flora between cows with mastitis and healthy cows, and the intestinal flora of dairy cows was transplanted into aseptic mice. The differences in the structure and function of this flora are also magnified to form a sharp contrast. The results showed that the structure of healthy intestinal flora had a direct effect on maintaining the healthy state of dairy cows. Probiotics could regulate the intestinal microflora of dairy cows, which could not be ignored to maintain the health of dairy cows, prevent and relieve mastitis.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S858.23
本文編號：2468069
[Abstract]:Mastitis is a high incidence in dairy cattle farms and easy to recur after being cured by antibiotics. The recurrence of the disease led to a continuous decline in milk production, nipple necrosis, and thus elimination of milk production in cows. Mastitis is also a common disease among lactating women. Traditional antibiotics treat neonatal nervous system damage. In the treatment of mastitis, because of its negative effects, antibiotics are no longer suitable for abuse in mastitis treatment. It is necessary to speed up the development of new and effective antibiotic substitutes for the treatment or prevention of mastitis. Probiotics have been successfully applied in the prevention and treatment of many diseases. Therefore, whether probiotics is really effective in the treatment and prevention of mastitis is an interesting research direction. Probiotics play a key role in the host intestine, so whether the intestinal flora is associated with mastitis is the first question whether probiotics can be used in the treatment of mastitis. In this paper, healthy cows and cows with mastitis were used to quantitatively study the common bacteria and bacteria in the intestinal microflora of healthy cows and mastitis cows. The results of correlation analysis showed that between the two groups of cows, the results of correlation analysis showed that there was no significant difference between the two groups of cows. The difference between the two groups was the largest by L.buchneri, with 27.32%, followed by L.sake, which reached 9.64%. Lactating aseptic mice were selected as animal models. Mastitis cow feces and healthy cow feces were transplanted into aseptic mice, and a group of mice transplanted mastitis cow feces were selected for probiotics intervention. Through microflora transplantation, breast tissue and other immune indexes of lactating mice were monitored. To investigate whether the mammary tissue of the mice transplanted with healthy cow feces and the mice transplanted with mastitis cow feces and the mice treated with probiotics at the same time had inflammation and the degree of inflammation. The results showed that mice transplanted with mastitis cow feces showed significant inflammation in breast tissue during 25 days of microflora transplantation, but no significant inflammation was found in mice transplanted with cow feces from healthy cows. The inflammation of mice treated with probiotics was weaker than that of mice treated with probiotics. After histopathological examination, three groups of mice fecal samples and two groups of cow fecal samples were collected and sequenced for metagenomic sequencing and 16s rRNA gene full-length sequencing. The results showed that the structure of intestinal flora was changed after it was transplanted into mice, but in the group of mice transplanted with cow feces, the structure change of intestinal flora was smaller, and it was more inclined to the structure of intestinal flora of cow in healthy group. Two groups of mice transplanted mastitis cow feces, one group of mice treated with probiotics, the intestinal microflora structure of the two groups of mice was smaller, but with mastitis cow intestinal flora structure change is greater. At the same time, the abundance of bacteria producing acetic acid, butyric acid and succinic acid was higher in the intestinal flora of healthy cows and mice transplanted with their feces. The full-length sequencing of 16s rRNA gene showed that the content of Lactobacillus in the feces of transplanted healthy cows was higher. From the results of macro-genome sequencing and full-length 16s rRNA gene sequencing, it was found that there were differences in the structure and function of intestinal flora between cows with mastitis and healthy cows, and the intestinal flora of dairy cows was transplanted into aseptic mice. The differences in the structure and function of this flora are also magnified to form a sharp contrast. The results showed that the structure of healthy intestinal flora had a direct effect on maintaining the healthy state of dairy cows. Probiotics could regulate the intestinal microflora of dairy cows, which could not be ignored to maintain the health of dairy cows, prevent and relieve mastitis.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S858.23
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