基于响应面法优化饲用粪肠球菌发酵培养基

为了实现粪肠球菌(Enterococcus faecalis)E_(XW)27的高密度培养,对健康仔猪肠道分离的粪肠球菌E_(XW)27发酵培养基进行响应面优化,以MRS为基础培养基,活菌浓度为评价指标,对发酵培养基中碳氮源、微量元素、缓冲盐体系以及促生长因子等通过单因素试验和响应面试验优化高密度发酵培养的培养基配方,确定最佳活菌浓度时的培养基组成。结果显示:蔗糖57.3 g/L、蛋白胨45 g/L、磷酸氢二钾3.75 g/L、柠檬酸铵3.5 g/L、乙酸钠6.25 g/L、硫酸镁1.5 g/L、硫酸锰0.45 g/L、组氨酸1.4 g/L、维生素C 0.01 g/L、碳酸钙10 g/L。发酵液最高活菌数达到1.03×10~(10) CFU/mL,是相同条件下MRS培养基中活菌数的10.61倍,菌体浓度显著提高,具有实际应用价值。     

粪肠球菌R-WL发酵培养基的优化

本试验旨在优化培养基提高粪肠球菌(Enterococcus faecalis)R-WL发酵液的菌体密度,为高效、优质的微生态制剂产品研制奠定基础。首先通过生长曲线的测定、培养条件和培养基成分的单因素试验,确定菌种RWL的最适培养条件为:时间14 h、pH 8.5、温度37℃、接种量3%,最适碳源为乳糖,浓度为10 g/L,最适氮源为蛋白胨+酵母膏,浓度为30 g/L。然后通过Plackett-Burman试验分析培养基中对粪肠球菌发酵影响最重要的主要因素为蛋白胨、乙酸钠和柠檬酸二铵,再通过最陡爬坡试验获得主要因素的最适范围,最后通过Box-Behnken试验和响应面分析得到主要因素的最适添加量。结果表明:最优发酵培养基配方为:乳糖10 g/L、酵母膏15 g/L、蛋白胨12.18 g/L、乙酸钠4.5 g/L、柠檬酸二铵4.5 g/L、硫酸镁0.58 g/L、硫酸锰0.25 g/L、磷酸氢二钾2 g/L、轻质碳酸钙3 g/L,初始pH 8.5。通过试验验证,优化后发酵培养基的发酵菌液活菌数可达(6.44±0.10)×109cfu/mL,比原MRS培养基活菌数提高了71.28%。     

屎肠球菌发酵工艺优化及菌粉的制备

为应用高活性的屎肠球菌菌粉制作畜禽饲料添加剂,试验对屎肠球菌的发酵培养基、发酵时间、喷粉载体等发酵和制粉条件进行优化。结果表明：最优发酵培养基选用32.50 g/L低蛋白乳清粉、30.00 g/L葡萄糖、2.00 g/L酵母提取物、5.00 g/L蛋白胨、0.15 g/L硫酸镁、0.03 g/L硫酸锰、2.00 g/L磷酸氢二钾,发酵时间为15 h,喷粉载体选用13%扶态粉、2%白炭黑作为保护剂制备菌粉。该方法制备的屎肠球菌菌粉中活菌数约3.46×10⁸CFU/g,可添加于畜禽饲料。     

屎肠球菌益生作用研究概述

屎肠球菌是一种益生菌,可以通过吸收动物体内的养分来维持体内肠道菌群的平衡,同时也可以调节动物小肠绒毛的发育,增强其抗病能力等.本文对屎肠球菌的生物学特性、调节胃肠道菌群作用、营养调控作用、益生作用机理、作为饲料添加剂的独特优势以及在药品和食品中的应用等进行了综述,以期为广大同行提供有益参考.     

5 L发酵罐中粪肠球菌发酵工艺参数优化

本试验旨在通过单因素试验及Box-Behnken响应面试验设计法优化饲用粪肠球菌EXW 27发酵工艺,探索搅拌速度、中和剂、pH、接种量、初始糖含量、通气方式以及补料工艺等对菌体在发酵罐内生长的影响。优化菌体小试发酵工艺为:将种子液以3%(V/V)接种于装液量为70%的5 L小罐,发酵温度37℃,搅拌转速165 r/min,通空气保压0.02 Mpa,自动流加12.5%的氨水控制pH在5.5,当pH高于5.5流加葡萄糖补料发酵18 h左右结束,在此最佳条件下所得发酵液的活菌浓度约为3.799×10^10 cfu/mL,比优化前提高了近3倍,为以后工业化生产提供参考。     

鲤鱼源粪肠球菌增菌培养基某的优化

利用SAS软件研究鲤鱼源粪肠球菌(Enterococcus fae clis)YL-1增茵培养基,首先,通过Plackett-Burman试验设计在培养基的各个因子中筛选出对YL-1生长有利的重要因子,然后在Box-Behnken试验设计下确定最大响应值,并通过优化的结果验证.结果表明预测值和实际值具有良好的拟合性,优化模型可靠,确定YL-1最佳培养基为:蛋白胨1.365%、牛肉膏1-25%、酵母浸粉0.625%、麦芽糖2.5%、吐温-80 0.1%、KH2PO4 0.2%、柠檬酸三铵0.16%、MgSO4 0.25%、初始pH值6.4.培养优化后YL-1的茵体浓度达1.71×10.CFU/ml.     

响应面法优化饲用屎肠球菌QW256增殖培养条件

为明确具有益生潜力的屎肠球菌QW256最适培养基与培养条件,利用Plackett-Burman试验、最陡爬坡试验、中心组合设计试验以及响应面法探究不同碳源、氮源、营养因子以及培养条件对菌株屎肠球菌QW256活菌数的影响。结果表明：屎肠球菌QW256菌株最佳增殖培养条件为接种量5%、培养温度37℃、初始pH 7.8;最适培养基成分为糖蜜24.7 g/L、酵母粉10 g/L、玉米浆16 g/L、柠檬酸0.25 g/L、柠檬酸钠5.1 g/L、硫酸镁0.6 g/L及精氨酸0.2 g/L。在优化后的培养基和培养条件下培养17 h时活菌数达1.12×10¹⁰CFU/mL,为其工业化应用提供了试验依据和技术支持。     

屎肠球菌及其在仔猪生产上的研究现状

屎肠球菌不仅具有优良的生物学特性,还可以调节机体免疫功能,提高采食量及饲料转化率,降低腹泻率,改善肠道内环境,促进动物健康,是畜牧生产及食品工业首选使用的益生菌菌种之一。文章主要综述了屎肠球菌的生物学特性及其在仔猪生产上的应用、目前存在的问题及今后的发展方向。     

微胶囊化屎肠球菌及其特性研究

试验采用发酵前包被的工艺,对所筛得屎肠球菌进行微胶囊化,并对微囊化菌体的特性进行了研究。结果表明:所制得微胶囊粒径大小合适,形态好;微囊化培养的屎肠球菌对致病型大肠杆菌K88、K99、金黄色葡萄球菌、巴氏杆菌及沙门氏菌都有较强的抑制作用,其中对大肠杆菌K99的抑菌效果最好;与游离培养状态下相比,微囊化培养的屎肠球菌具有更好的生长优势,更好的抵御高铜、模拟胃液的能力(P<0.01);储存试验表明,4℃条件下,储藏2个月,活菌数基本没有下降。由此可见,微胶囊包被能明显提高屎肠球菌在极端环境下的存活率。     

乌鲁木齐市宠物源粪肠球菌和屎肠球菌耐药性及耐药基因检测

为了解乌鲁木齐市宠物源粪肠球菌和屎肠球菌对临床常用抗菌药物的耐药性及其耐药基因携带情况,在乌鲁木齐市多家宠物医院、警犬基地、犬养殖基地采集宠物(犬和猫)直肠粪样449份进行粪肠球菌和屎肠球菌的分离鉴定,通过琼脂稀释法测定10种抗菌药物的最小抑菌浓度,利用PCR方法检测11种耐药基因.结果分离鉴定出宠物源粪肠球菌和屎肠球...     

枯草芽孢杆菌Bacillus subtilis BS1发酵培养基的优化

为提高枯草芽孢杆菌BS1发酵液中芽孢含量,通过单因素试验、正交试验及回归正交组合试验优化其发酵培养基,最终确定培养基配方为:麦芽浸粉1 g/l、黄豆粉15 g/l、CaCO36.68 g/l、MnSO40.4 g/l、MgCl22 g/l,初始pH值7.0。使用该配方,在37℃、180 r/min条件下于恒温摇床中振荡培养48 h,枯草芽孢杆菌BS1菌株发酵液中芽孢含量可达8.55×1010cfu/ml。     

一株致病性粪肠球菌的分离与鉴定

从1羽发病雏鸡中分离到1株溶血性革兰氏阳性球菌。初步分离鉴定后经攻毒试验能够造成小白鼠和雏鸡死亡,具有致病性。药敏试验表明,该菌对多种药物均表现耐药性,仅对庆大霉素、青霉素、氧氟沙星敏感。通过BLAST软件分析该菌株的16S rRNA序列,确定该致病性细菌为粪肠球菌。     

Vancomycin for multi-drug resistant Enterococcus faecium cholangiohepatitis in a cat

A 12-year-old, neutered male domestic shorthair cat was evaluated with a life-long history of intermittent, predominantly small bowel diarrhea and a 3 day history of hematochezia. At presentation, the cat had increased liver enzyme activities and an inflammatory leukogram. Histopathology demonstrated inflammatory bowel disease (IBD), cholangiohepatitis and pancreatitis. The cholangiohepatitis was associated with a multi-drug resistant Enterococcus faecium. Gallbladder agenesis was also documented. Treatment with vancomycin was safely instituted for 10 days. Clinical signs resolved, however, cure of the bacterial cholangiohepatitis was not achieved. The risk of vancomycin resistant enterococci (VRE) in human and veterinary medicine is discussed.     

地衣芽孢杆菌TS-01发酵培养基的优化

通过正交实验对地衣芽孢杆菌TS-01的发酵培养基在摇瓶中进行了优化。研究发现:当培养基中葡萄糖为20g/l、豆饼粉为70g/l、玉米浆为7g/l时,发酵液中活菌数高于其它配比的培养基,达9.6×108cfu/ml。     

Solid-state fermentation of corn-soybean meal mixed feed with Bacillus subtilis and Enterococcus faecium for degrading antinutritional factors and enhancing nutritional value

Background: Corn and soybean meal(SBM) are two of the most common feed ingredients used in pig feeds.However, a variety of antinutritional factors(ANFs) present in corn and SBM can interfere with the bioavailability of nutrients and have negative health effects on the pigs. In the present study, two-stage fermentation using Bacillus subtilis followed by Enterococcus faecium was carried out to degrade ANFs and improve the nutritional quality of corn and SBM mixed feed. Furthermore, the microbial composition and in vitro nutrient digestibility of inoculated mixed feed were determined and compared those of the uninoculated controls.Results: During the fermentation process, B. subtilis and lactic acid bacteria(LAB) were the main dominant bacteria in the solid-state fermented inoculated feed, and fermentation produced a large amount of lactic acid(170 mmo L/kg),which resulted in a lower pH(5.0 vs. 6.4) than the fermented uninoculated feed. The amounts of soybean antigenic proteins(β-conglycinin and glycinin) in mixed feed were significantly decreased after first-stage fermentation with B. subtilis. Inoculated mixed feed following two-stage fermentation contained greater concentratioin of crude protein(CP), ash and total phosphorus(P) compared to uninoculated feed, whereas the concentrations of neutral detergent fiber(NDF), hemicellulose and phytate P in fermendted inoculated feed declined(P 0.05) by 38%, 53%, and 46%,respectively. Notably, the content of trichloroacetic acid soluble protein(TCA-SP), particularly that of small peptides and free amino acids(AA), increased 6.5 fold following two-stage fermentation. There was no difference in the total AA content between fermented inoculated and uninoculated feed. However, aromatic AAs(Phe and Tyr) and Lys in inoculated feed increased, and some polar AAs, including Arg, Asp, and Glu, decreased compared with the uninoculated feed. In vitro dry matter and CP digestibility of inoculated feed improved(P 0.05) compared with the uninoculated feed.Conclusions: Our results suggest that two-stage fermentation using B. subtilis followed by E. faecium is an effective approach to improve the quality of corn-soybean meal mixed feed.     

屎肠球菌对仔猪肠道健康的影响

屎肠球菌是仔猪生产中常用的益生菌之一。该益生菌不仅能够在仔猪的胃肠道中存活,并维持仔猪肠道菌群平衡,还可以调节仔猪的小肠绒毛发育、养分吸收、免疫功能以及抗衣原体、轮状病毒和鼠伤寒沙门氏菌感染的能力,对仔猪肠道健康有重要影响。本文就上述研究进展进行综述。