补喂复合益生菌对3～6月龄伊犁马粪便纤维含量、挥发性脂肪酸浓度和血浆细胞因子水平的影响

试验旨在研究补喂复合益生菌对3～6月龄伊犁马粪便纤维含量、VFA浓度和血浆细胞因子水平的影响。选取体重相近(90.23kg±11.02kg)、健康状况良好的3月龄伊犁马哺乳马驹10匹,随机分为2组,每组5匹,分别为对照组和试验组。在分阶段补喂等量营养水平的精料补充料和苜蓿干草的基础上,试验组每匹马驹每天补喂0.075g(2×10¹¹ CFU/g)枯草芽孢杆菌和0.133g(3×10¹⁰ CFU/g)植物乳杆菌复合菌,进行为期90d的饲养试验。结果显示,补喂复合益生菌能够显著或极显著降低马驹粪便中酸性洗涤纤维(ADF)和淀粉的含量(P<0.05;P<0.01),分别比对照组低3.50%和11.32%;显著或极显著提高马驹粪便中乙酸和丙酸的浓度(P<0.05;P<0.01),分别比对照组高14.65%和64.29%;极显著提高血浆中肿瘤坏死因子α(TNF-α)的水平(P<0.01),比对照组高16.17%。由此可见,补喂复合益生菌能够降低马驹粪便中纤维的含量、提高挥发性脂肪酸(VFA)的浓度,增加3～6月龄伊犁马血浆细胞因子水平。     

动物乳杆菌发酵液的体外抑菌试验

采用牛津杯法测定了动物乳杆菌发酵液及除酸、除过氧化氢后的发酵液对各种指示菌的抑制作用。研究结果显示：动物乳杆菌原始发酵液的抑菌效果最好;除酸后的发酵液的抑菌效果有一定变化,抑菌谱减小;而除过氧化氢后的发酵液的抑菌谱与除酸后的没有变化。     

Screening of Probiotics Inhibiting E. coli K88 in vitro

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.     

猪轮状病毒Vp4全基因与乳酸菌非抗性表达载体的重组及在大肠杆菌中的表达

以猪A组轮状病毒mRNA为模板，应用优化的反转录聚合酶链式反应（RT—PCR）技术，扩增了2331 bp的Vp4全长基因，通过T-A克隆技术，将PCR产物克隆至克隆载体pGEM-T中，构建质粒pGEM-T-Vp4。以SalⅠ和KpnⅠ双酶切pGEM—T—Vp4．和以胸苷酸合成酶基因（thymidylate synthase，thyA）为选择压力的非抗生素抗性的穿梭表达载体pW425t，并将纯化的Vp4基因亚克隆至表达载体pW425t中，构建出可以在乳酸菌与大肠杆菌之间穿梭表达的原核表达重组质粒pW425t-Vp4。将pW425t-Vp4转化至thyA基因缺陷型的大肠杆菌感受态E．coli X13中，通过生长功能弥补筛选阳性克隆，SDS-PAGE分析，可见约87.67Ku融合蛋白。Western blot分析表明，该蛋白具有与轮状病毒多克隆抗体的反应原性，研究结果为pW425t-Vp4在乳酸菌受体菌株中表达提供科学依据。     

嗜酸乳杆菌生产共轭亚油酸的发酵条件研究

[目的]为提高微生物法发酵合成共轭亚油酸(CLA)的产量,优化嗜酸乳杆菌生产共轭亚油酸的发酵条件.[方法]试验以亚油酸为底物,利用嗜酸乳杆菌合成共轭亚油酸,选取对CLA的生成量影响显著的几个因素:底物浓度、接种量、pH、发酵时间、发酵温度进行单因素试验,并在此基础上进行正交试验优化发酵条件.[结果]单因素试验得到的最优发酵条件为底物浓度1.0 mg/ml、接种量4％、初始pH 5.5、发酵时间48 h、发酵温度37℃.正交试验发现,底物浓度、发酵时间、发酵温度显著影响CLA的合成产量.最终得到的最优发酵条件为底物浓度为1.5 mg/ml、接种量2％、初始pH 5.5、发酵时间36 h、发酵温度33℃.用气质联用法对产物进行定性分析,证明产物是c-9,t-11 CLA.此时,CLA的产量为91.54 μg/ml.[结论]试验优化了共轭亚油酸的发酵条件,为更有效率地利用嗜酸乳杆菌生产共轭亚油酸提供了理论依据.     

嗜酸乳杆菌La-1产细菌素发酵条件的研究

合适的发酵条件能够有效地提高嗜酸乳杆菌La-1细菌素的产量。在已经确定碳源、氮源和复合维生素的基础上,采用正交试验的方法研究了以MRS为基础的发酵培养基中各营养成分的相互协同作用,10L发酵罐试验证实,优化后的培养基和各项优化技术参数能使细菌总数提高了53.6%,细菌素抗菌活性提高了35.3%,抑菌圈达到了25mm以上。     

Effects of ethanol,molasses and Lactobacillus plantarum on fermentation characteristics and aerobic stability of total mixed ration silages

This study was conducted to evaluate the effects of Lactobacillus plantarum, molasses and/or ethanol on fermentation quality and aerobic stability of total mixed ration (TMR) silage, which is widely used in dairy cow diets at mid‐to‐late lactation in Tibet. TMR was treated with ethanol (E), molasses (M), Lactobacillus plantarum(L), ethanol+molasses (EM), ethanol+Lactobacillus plantarum (EL) plus control. After 45 d of ensiling, inoculant significantly (P < 0·05) increased lactic acid (LA) concentration and decreased pH, ammonia nitrogen (AN) concentration, and aerobic bacterial and yeast counts, compared to control. After the first 3 d of aerobic exposure, LA for silages without ethanol started to decrease, while LA for E silages almost remained unchanged till the end of the aerobic exposure period. The pH in TMR silages without ethanol gradually increased, while that for E and EL remained about 4·60 and 4·00, respectively, and EL showed the lowest pH among six silages over the course of aerobic exposure. Aerobic bacterial counts in control, M and EM silages were significantly higher (P < 0·05) than those in E, L and EL, and yeast counts in E and EL silages were significantly lower (P < 0·05) than those in other silages after 9 d of aerobic exposure. The results suggest that inoculation with L. plantarum was more effective in altering fermentation characteristics than adding molasses, while ethanol showed a potential to protect TMR silages from pH increase and delayed the growth of aerobic bacteria and yeast either alone or in combination with L. plantarum.     

嗜酸乳酸菌对吉富罗非鱼生长、非特异性免疫酶活性和肠道菌群的影响

选用初始体重为2.43(±0.03)g的吉富罗非鱼(GIFT Oreochromis niloticus),随机分为5组,分别投喂含0、8x105、4x106、2x107、1.5×108 CFU/g嗜酸乳酸菌(Lactobacillus acidophilus)的试验饲料,记为G0 、G1、G2、G3和G4.养殖42 d后,测定罗非鱼的生长指标、体成分、血清非特异性免疫相关酶活性和肠道细菌数量.结果显示,G3和G4的增重率(WGR)、特定生长率(SGR)和蛋白质效率(PER)均高于G0,但未到达显著水平;饲料系数(FC)以G3最低,各组之间差异不显著.饲料中添加乳酸菌对罗非鱼全鱼水分和灰分含量没有显著性影响,粗蛋白和粗脂肪含量有不同程度的升高,但差异不显著.与G0相比,添加嗜酸乳酸菌可提高罗非鱼血清超氧化物歧化酶(SOD)、碱性磷酸酶(AKP)和酸性磷酸酶(ACP)活性,其中G3组SOD活性显著升高.饲料中添加嗜酸乳酸菌能降低罗非鱼肠道细菌总数,其中G2和G3显著低于G0;G2、G3和G4罗非鱼肠道中大肠杆菌数与G0相比显著降低;罗非鱼肠道乳酸菌数量随嗜酸乳酸菌添加水平的增加呈现升高的趋势,但差异未达到显著性水平;各组罗非鱼肠道双歧杆菌数量差异不显著.结果表明,饲料中添加嗜酸乳酸菌可一定程度促进吉富罗非鱼的生长,提高血清SOD和AKP活性,降低肠道中细菌总数,抑制大肠杆菌生长.     

玉米浸泡液中噬淀粉乳杆菌的分离与鉴定

从玉米浸泡液中分离出３株细菌，这些细菌为兼性厌氧，革兰氏阳性，无运动性，不形成芽胞的杆菌，呈单在或短链状排列．无过氧化氢酶．４５℃生长，１５℃不生长，发酵纤维二糖、水杨苷，不发酵阿拉伯糖、棉子糖、鼠李糖，水解淀粉．这些细菌与Ｌ．Ｋ．Ｎａｋａｍｕｒａ（１９８１）所报道的噬淀粉乳杆菌（Ｌａｃｔｏｂａｃｉｌｌｕｓａｍｙｌｏｖｏｒｕｓ）的性状完全一致，证明由玉米制淀粉的过程中有噬淀粉乳杆菌参与．     

Lactobacillus parafarraginis ZH1 producing anti‐yeast substances to improve the aerobic stability of silage

Enhancing the aerobic stability of silage is very important in silage production. The objective of this study was to compare the roles of a new bacterial strain Lactobacillus parafarraginis ZH1 with Lactobacillus plantarum Chikuso‐1 (LP) and Lactobacillus buchneri NCIMB 40788 (LB) in improving the aerobic stability of oat silage and identify the anti‐yeast substances produced by them. After ensiled for 45 days, either inoculated silages or control silage were of pH values lower than 4.2. The control and LP inoculated silage had poorer aerobic stability than LB and ZH1 inoculated ones (p < 0.05). ZH1 inoculated silage produced more acetic acid, benzoic acid and hexadecanoic acid than LB inoculated one and had the best aerobic stability (p < 0.05). In MRS medium, strain ZH1 produced more hexadecanoic acid than LP and LB strains, more benzoic acid than strain LP (p < 0.05), and the equal amount of benzoic acid to LB (p > 0.05). In addition, benzoic acid and hexadecanoic acid had low minimal inhibitory concentrations to target yeasts in the pure culture. In conclusion, L. parafarraginis ZH1 had stronger anti‐yeast potential and more effectively improved the aerobic stability of silage than other strains used in this study.