复合乳酸菌制剂的耐受性和抑菌性研究

本试验将四株饲用乳酸菌,制成复合乳酸菌制剂,通过单因素试验设计,以存活率为指标,研究复合微生态产品对高温、酸、胆盐的耐受性,采用琼脂稀释法、牛津杯法分别测定复合乳酸菌制剂对不同种类饲用抗生素的耐受性及对致病菌的抑制作用。结果表明:复合乳酸菌产品在温度65℃处理2.5、5、7.5 min以及85℃处理2.5 min对其存活率没有影响,但当85℃处理5 min,存活率下降至48%;p H为3时处理90 min存活率高达80.54%。对0.3%猪胆盐的耐受性较好,处理1 h存活率为73.52%;对抗生素(药物饲料添加剂)杆菌肽锌、亚甲基水杨酸杆菌肽耐受性最强,对吉他霉素的耐受性最差;可抑制致病菌株大肠埃希氏菌的生长。体外评价试验结果表明,复合乳酸菌制剂具有很好的益生特性,可进一步研究其在动物上的应用效果。     

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。     

被膜态与浮游态猪源乳杆菌上清液抑菌活性比较分析

试验旨在比较分析猪源约氏乳杆菌L-76和嗜淀粉乳杆菌L-102被膜态和浮游态下其上清液对病原菌的抑菌活性。采用结晶紫染色法测定L-76和L-102的生物被膜形成能力,研究其被膜态和浮游态乳杆菌上清液及上清液与不同影响因素作用后的抑菌活性,并利用扫描电镜观察其两种状态下的上清液对指示菌形态的影响。结果显示,L-76、L-102乳酸菌具有较强的生物被膜形成能力,被膜态和浮游态乳杆菌L-76、L-102具有良好的抑菌活性,其上清液经蛋白酶作用后,被膜态L-76和L-102上清液的抑菌圈直径比浮游态明显变小;经过氧化氢酶和不同温度作用后,两种状态的乳杆菌上清液的抑菌活性无明显变化（P>0.05）,但在pH 3.0作用下,两种状态的乳杆菌上清液的抑菌效果最好;扫描电镜结果显示,被膜态乳杆菌上清液对金黄色葡萄球菌的形态影响略大。综上所述,两种状态下乳酸杆菌上清液中均含有抑菌物质,且被膜态乳酸杆菌上清液中的抑菌物质含量略多或活性略高。     

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.     

Research Lactobacillus rhamnosusFermented Herbal and Bacillus subtilis Complex on Immune Performance,E.coli Infection of Broiler

This experiment was designed to study the complex of Lactobacillus rhamnosus fermented herbal and Bacillus subtilis on White Feather broiler immunity performance and impact of Escherichia coli infection.360 one-day-old broiler chickens were randomly divided into 3 groups with 4 replicates in each group and 30 chickens per replicate.The pretrial period lasted for 7 d,and the experiment lasted for 35 d.The chickens in the group Ⅰ(positive control group) and group Ⅱ(negative control group)were all only fed a basal diet,group Ⅲ was test group,by additive 1% fermented herbal preparations,groups Ⅱ and Ⅲ were intraperitoneal injection of 1 mL E.coli at 35 d,broiler mortality,immune organ index,cecalmicroflora content,immunoglobulin levels,IL-2 and IL-6 content were tested.The results showed that injection of E.coli caused massive death of chickens,group Ⅱtook up to 75.00%,it was significantly higher than groups Ⅰ and Ⅲ (P < 0.05),the mortality in group Ⅲ was significantly lower than group Ⅱ(P < 0.05),was only 23.33%.Injection of E.coli maked spleen index and thymus index of group Ⅱincreased significantly (P < 0.05),the spleen index and thymus index of groups Ⅰ and Ⅲ were no significant difference (P > 0.05).E.coli counts was significantly decreased after injectionin group Ⅲ (P < 0.05),but the number of intestinal Lactobacilli of group Ⅲ was significantly increased (P < 0.05),and inhibited the propagation of E.coli,the counts of E.coli in groups Ⅰ and Ⅲ were no significant difference (P > 0.05).At 42 d,the sIgA of the intestinal fluid in group Ⅲ were higher than that of groups Ⅰ and Ⅱ with 11.99% and 36.56%,respectively(P < 0.05).The serum IgG concentrations of group Ⅲ was higher than that of groupsⅠand Ⅱwith 14.68% and 28.15%,respectively(P < 0.05).At 42 d,the IL-2 content of group Ⅱ was the lowest,it was significantly lower than group Ⅲ(P < 0.05),the IL-6 of group Ⅲ was significantly lower than group Ⅱ(P < 0.05).     

Lactobacillus rhamnosus GG SpaC pilin subunit binds to the carbohydrate moieties of intestinal glycoconjugates

Lactobacillus rhamnosus GG (LGG) is a well‐established probiotic strain. The beneficial properties of this strain are partially dependent on its prolonged residence in the gastrointestinal tract, and are likely influenced by its adhesion to the intestinal mucosa. The pilin SpaC subunit, located within the Spa pili structure, is the most well studied LGG adhesion factor. However, the binding epitopes of SpaC remain largely unknown. The aim of this study was to evaluate the binding properties of SpaC to the carbohydrate moieties of intestinal glycoconjugates using a recombinant SpaC protein. In a competitive enzyme‐linked immunosorbent assay, SpaC binding was markedly reduced by addition of purified mucin and the mucin oligosaccharide fraction. Histochemical staining revealed that the binding of SpaC was drastically reduced by periodic acid treatment. Moreover, in the surface plasmon resonance‐based Biacore assay, SpaC bound strongly to the carbohydrate moieties containing β‐galactoside at the non‐reducing terminus of glycolipids. We here provide the first demonstration that SpaC binds to the oligosaccharide chains of mucins, and that the carbohydrate moieties containing β‐galactoside at the non‐reducing termini of glycoconjugates play a crucial role in this binding. Our results demonstrate the importance of carbohydrates of SpaC for mucus interactions.     

多型FMDV VP1表位嵌入型载体pMG-SLPMultiVP1的构建及其表达产物鉴定

构建出表达载体pMG-SLPMultiVP1,并对其进行双酶切鉴定及PCR鉴定。将阳性重组质粒转入到发酵乳杆菌中进行表达,并对表达产物进行SDS-PAGE、Western blot和IFAT鉴定。多型FMDV VP1表位嵌入的SLPMultiVP1融合基因成功克隆入穿梭质粒pMG36e中,其表达产物大小约为35ku,部分表达产物位于发酵乳杆菌表面。成功构建了一个乳杆菌嵌入型表达载体pMG-SLPMultiVP1,为以发酵乳杆菌为活菌载体的表位疫苗研究奠定了基础。     

同源短乳杆菌对鸡肠黏膜SIgA分泌的影响

为了了解短乳杆菌对机体免疫力的影响,用MRS培养基从鸡的消化道分离到乳酸杆菌,对该菌的形态特征、培养特性、生理生化特性等进行试验,证明该菌为短乳杆菌.用短乳杆菌饲喂本地肉仔鸡后检测其肠内容物SIgA滴度.结果显示,该菌对肠道粘膜有较强的粘附力,能提高鸡肠黏膜的SIgA滴度.本试验表明,短乳杆菌能显著提高机体的免疫力.     

利用16S rRNA全长序列鉴定植物乳杆菌

从标明含有嗜酸乳杆菌的活菌胶囊中分离到一株乳杆菌Lal菌株，利用16S rRNA全序列分析法和传统分类法对Lal菌株进行分类鉴定表明：Lal菌株属于植物乳杆菌种新株系，该菌株现在已经被中国科学院北京微生物所菌种保藏中心收录保存，编号为ASl．2986。     

4株酸奶噬菌体的分离

从上海光明乳业股份有限公司车间发酵罐中分离出4株酸奶噬菌体。1号噬菌体头部呈六角形，平均直径为95.1nm，头部剖面平均面积为8184．8nm^2，尾部的平均直径为18．3nm，平均长度为403．4nm；2号噬菌体头部呈六角形，平均直径为85．4nm，头部剖面平均面积为8046．2nm^2，尾部的平均直径为23．3nm，平均长度为549．4nm；3号噬菌体头部呈六角形，平均直径为95．5nm，头部剖面平均面积为9792．3nm^2，尾部的平均直径为19．8nm，平均长度为450．5nm；4号噬菌体头部呈六角形，平均直径为119．3nm．头部剖面平均面积为12586．1nm^2，尾部的平均直径为23．6nm，平均长度为520．5nm。