本文選題：益生菌　切入點(diǎn)：發(fā)酵優(yōu)化　出處：《蘭州交通大學(xué)》2015年碩士論文

【摘要】：濫用抗生素嚴(yán)重地影響著畜產(chǎn)品的安全和質(zhì)量,尋找一種抗生素替代品成為當(dāng)前畜牧養(yǎng)殖急需解決的問題。復(fù)合微生物飼料添加劑來源于腸道正常菌群,能夠通過優(yōu)勢(shì)菌群、生物拮抗等作用改善腸道微生物環(huán)境。調(diào)整并維持腸道微生物平衡,增強(qiáng)免疫能力,促進(jìn)營(yíng)養(yǎng)物質(zhì)的消化吸收,起到促生長(zhǎng)和防病的作用。因此,益生菌復(fù)合制劑已成為取代抗生素的首選開發(fā)研究對(duì)象。本研究采用耐酸和耐膽鹽試驗(yàn)、選擇性培養(yǎng)等方法,成功地從斷奶羔羊新鮮糞便中分離純化出一株具有良好耐受模擬胃酸和模擬膽鹽環(huán)境的菌株,經(jīng)鑒定為乳酸菌R1。通過單因素試驗(yàn)、正交優(yōu)化試驗(yàn)、添加菌體促生長(zhǎng)因子試驗(yàn)等對(duì)乳酸菌進(jìn)行發(fā)酵培養(yǎng)基和培養(yǎng)條件的研究。同時(shí)對(duì)本實(shí)驗(yàn)室保藏的一株枯草芽孢桿菌與一株釀酒酵母進(jìn)行了培養(yǎng)基和培養(yǎng)條件的優(yōu)化。并對(duì)乳酸菌R1進(jìn)行了微膠囊包埋,采用真空恒溫干燥法將枯草芽孢桿菌發(fā)酵液制備干菌粉,采用自然風(fēng)干法將釀酒酵母固體發(fā)酵物干燥研磨制備干菌粉。最后將乳酸菌R1微膠囊、枯草芽孢桿菌干菌粉和釀酒酵母干菌粉復(fù)配。制備了復(fù)合微生物飼料添加劑。利用此制劑在羔羊上進(jìn)行了動(dòng)物活體實(shí)驗(yàn)驗(yàn)證。實(shí)驗(yàn)結(jié)果如下:(1)乳酸菌R1在初始為培養(yǎng)條件為:pH6.0、溫度34℃、靜置培養(yǎng)12-14小時(shí)、接種量4%時(shí)生長(zhǎng)狀態(tài)最好。最適培養(yǎng)基為:蛋白胨0.5%,葡萄糖2%,牛肉膏1%,酵母膏0.5%,吐溫80 0.1%,碳酸鈣0.2%,乙酸鈉0.5%,檸檬酸二銨0.2%,硫酸鎂0.02%,硫酸錳0.005%;增菌因子配比為:番茄汁2%,土豆汁3%,胡蘿卜汁6%,維生素C 0.2%;活菌數(shù)可以達(dá)到7.41×109 CFU/m L。乳酸菌R1微膠囊包覆率達(dá)到了90.5%同時(shí)具有較好的耐酸性、較好的腸溶性,在常溫儲(chǔ)藏90d存活率為79.5%,活菌數(shù)為3.2×109 CFU/g。(2)枯草芽孢桿菌在初始培養(yǎng)條件為:pH為7.0-7.5、溫度35℃、裝液量30mL/250、接種量7%、轉(zhuǎn)速為200r/min、培養(yǎng)17h生長(zhǎng)狀態(tài)最好。最適培養(yǎng)基為葡萄糖3%、酵母膏1.5%、磷酸氫二鈉0.2%、磷酸二氫鈉0.1%;菌體生物量達(dá)到7.90×109 CFU/m L。木屑添加量為15g/25m L,芽孢產(chǎn)率接近95%,制備的干菌粉活菌數(shù)為5.71×1010CFU/g.(3)釀酒酵母在培養(yǎng)條件為:搖床轉(zhuǎn)速150 r/min、溫度25℃,接種量1%、裝液量25m L/250m L、培養(yǎng)20h生長(zhǎng)狀態(tài)最好。最佳液體培養(yǎng)基為紅糖5%、酵母膏2%、硫酸銨0.1%、氯化鉀0.2%;酵母活菌數(shù)達(dá)到了2.38×108 CFU/mL固體培養(yǎng)基按麩皮∶糖蜜∶豆粕=7∶2∶1的比例配制,料水比1:0.6,用礦物質(zhì)鹽溶液拌料,培養(yǎng)時(shí)間60h,生物量最大值為8.73×109 CFU/g,比優(yōu)化前提高了231.8倍。(4)動(dòng)物活體試驗(yàn)表明:復(fù)合微生物飼料添加劑的使用,顯著地提高了羔羊的日增重,降低肉料比,降低了斷奶羔羊腹瀉發(fā)生率,其中對(duì)照組腹瀉發(fā)生率為26.7%,試驗(yàn)組腹瀉發(fā)生率僅為6%,羔羊腹瀉率降低了20.7%。本研究篩選得到較優(yōu)良的微生物飼料添加劑菌株,對(duì)其進(jìn)行發(fā)酵優(yōu)化,制劑增重防病效果顯著,為實(shí)際生產(chǎn)微生物飼料添加劑提供了一定的理論參考價(jià)值。
[Abstract]:The abuse of antibiotics seriously affect the quality and safety of animal products, to find a substitute for antibiotics has become the urgent problem to be solved. Animal husbandry compound microbial feed additives derived from normal intestinal flora, the dominant microflora, improve the intestinal microbial environment biological antagonistic action. Adjust and maintain the intestinal microflora balance, enhance immunity and promote digestion and absorption of nutrients, to promote growth and prevent disease. Therefore, probiotic combination has become a substitute for antibiotics preferred object of study. This study used acid and bile salts resistance test method of selective culture, successfully weaned lamb from fresh feces of isolated strains with good tolerance the simulation of gastric acid and bile salt environment simulation, and identified as Lactobacillus R1. by single factor test, orthogonal test, cell growth by adding Test of medium and culture conditions of the fermentation of lactic acid bacteria. Meanwhile a Bacillus subtilis strain of the yeast Saccharomyces cerevisiae and the optimization of medium and culture conditions of lactic acid bacteria. R1 was microencapsulated by vacuum drying, constant temperature method of Bacillus subtilis preparation of fermentation liquor dry bacterium powder, drying method of yeast fermentation solid preparation of dry grinding dry bacterium powder. The lactic acid bacteria R1 microcapsule, Bacillus subtilis bacteria and yeast powder dry dry bacterium powder compound. The composite microbial feed additives were prepared. The animal experiment in lamb by using this formulation. The experimental results are as follows: (1) R1 of lactic acid bacteria in the initial condition is: pH6.0, temperature 34 C, static culture for 12-14 hours, when the inoculation amount was 4%, the best growth state. The optimum medium was peptone 0.5%, glucose 2%, beef extract 1%, yeast extract 0.5%, Twain 800.1%, calcium carbonate 0.2%, 0.5% sodium acetate, ammonium citrate 0.2%, Magnesium Sulfate 0.02%, 0.005% manganese sulfate; enrichment factor ratio: Tomato Juice 2%, potato juice 3%, Fresh Carrot Juice 6%, vitamin C 0.2%; viable count can reach 7.41 * 109 CFU/m L. lactic acid bacteria R1 microencapsulation rate reached 90.5% and has good acid resistance, enteric and good storage in 90d survival rate was 79.5%, the living bacteria number is 3.2 * 109 CFU/g. (2) of Bacillus subtilis in initial culture conditions: pH 7.0-7.5, 35 degrees Celsius temperature, liquid volume 30mL/250. 7% inoculation quantity, speed 200r/min, 17h culture growth status of the best. The best medium was 3% glucose, 1.5% yeast extract, two sodium hydrogen phosphate 0.2%, 0.1% sodium dihydrogen phosphate; biomass reached 7.90 * 109 CFU/m L. sawdust was added into 15g/25m L, spore yield close to 95%, number of live bacteria was 5.71 bacteria powder preparation dry preparation * 1010CFU/g. (3) in yeast culture conditions: rotation speed of 150 r/min, temperature 25, inoculum size 1%, medium volume 25m L/250m L, 20h culture medium. The growth state of the best the best liquid is brown sugar 5%, yeast extract 2%, 0.1% ammonium sulfate, potassium chloride 0.2%; yeast bacteria number reached 2.38 x 108 CFU/mL solid medium by bran: molasses meal: =7: 2: 1 of the proportion, the ratio of material to water 1:0.6, with mineral salt solution mixing, fermentation time 60H, the maximum biomass was 8.73 * 109 CFU/g, 231.8 times more than that before optimization. (4) animal experiment table Ming: the use of composite microbial feed additives, and significantly improve the daily gain of lamb, decrease the ratio of feed to meat, reduce the incidence of diarrhea of weaned lamb, which controls the incidence of diarrhea was 26.7%, the incidence of diarrhea in experimental group was only 6%, lamb diarrhea rate decreased by 20.7%. in this study were obtained excellent. The fermentation of the feed additive strain has a significant effect on the weight gain and disease prevention. It provides a theoretical reference value for the actual production of microbial feed additives.

【學(xué)位授予單位】：蘭州交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S816.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳丹丹;顧勝華;張金娜;寶福凱;湯岳琴;吳曉磊;;腸道菌群對(duì)免疫系統(tǒng)的影響及其群落分析方法[J];應(yīng)用與環(huán)境生物學(xué)報(bào);2013年03期

2 華均超;張邦輝;周明;李金友;;屎腸球菌T013復(fù)合制劑在斷奶仔豬生產(chǎn)中的應(yīng)用研究[J];安徽農(nóng)業(yè)科學(xué);2012年11期

3 葉雪飛;阮輝;馮石開;朱承亮;何國(guó)慶;;發(fā)酵乳桿菌增殖培養(yǎng)基營(yíng)養(yǎng)因子優(yōu)化研究[J];食品科學(xué);2010年05期

4 代永剛;田志剛;南喜平;;乳酸菌及其生理功能研究的進(jìn)展[J];農(nóng)產(chǎn)品加工(學(xué)刊);2009年07期

5 張日俊;;微生物飼料添加劑的科學(xué)使用[J];飼料與畜牧;2007年06期

6 張董燕;季海峰;徐煒玲;;益生菌對(duì)動(dòng)物腸道微生物生態(tài)學(xué)影響的研究進(jìn)展[J];中國(guó)畜牧獸醫(yī);2007年03期

7 潘寶海;張建東;譙仕彥;;芽孢桿菌對(duì)畜禽生產(chǎn)性能的影響[J];飼料研究;2007年01期

8 張日俊;動(dòng)物微生態(tài)系統(tǒng)的生物防治和營(yíng)養(yǎng)免疫作用及微生物飼料添加劑的科學(xué)使用[J];飼料工業(yè);2005年16期

9 王桂瑛,劉燕;益生素的研究及應(yīng)用進(jìn)展[J];飼料博覽;2004年06期

10 黃慶飛,戴永恒,莫文偉,余桂華;EM菌技術(shù)在奶牛飼養(yǎng)中的應(yīng)用效果研究[J];廣西畜牧獸醫(yī);2004年03期

，

本文編號(hào)：1698013