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).     

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。     

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.     

低聚木糖的制备及其对益生菌体外增殖的作用

针对玉米芯微波消解-内切木聚糖酶水解制备低聚木糖的工艺,以低聚木糖的得率为主体评价指标,通过单因素实验时影响低聚木糖得率的微波消解过程和内切木聚糖酶水解过程的因素与水平进行研究,并考察获得的低聚木糖对益生菌的体外增殖作用.结果表明:玉米芯酶法制备低聚木糖的最佳工艺条件为微波处理压力1.6 MPa,微波处理时间5 min,内切木聚糖酶用量140 U·g-1,酶解时间6 h;在最适条件下,玉米芯酶解液中低聚木糖的得率为82.5%,质量浓度为11.02 g·L-1;低聚木糖对益生菌的体外增殖实验表明,低聚木糖添加量在0.2%和0.4%时,分别可以对乳酸杆菌和枯草芽孢杆菌起到最好的增殖作用,分别达到311%和183%,添加量进一步提高反而会抑制这2种菌的生长.     

鼠李糖乳杆菌益生素对猪生产性能及血清学指标的影响

[目的]为今后鼠李糖乳杆菌益生素的开发应用奠定基础.[方法]对照组猪饲喂基础日粮,试验Ⅰ组和Ⅱ组分别饲喂添加2和4ml/头鼠李糖乳杆菌益生素(RLB)的基础日粮,研究鼠李糖乳杆菌益生素对猪生产性能及部分血清学指标的影响.[结果]试验Ⅰ组和Ⅱ组妊娠母猪的死产、畸形、木乃伊和弱仔猪的总数量比对照组减少36.36％和54.55％,且平均初生重提高5.38％和10.00％;试验Ⅰ组和Ⅱ组断奶仔猪的平均重量比对照组提高1.31％和2.96％;与对照组相比,试验Ⅰ组和Ⅱ组断奶仔猪的平均日增重分别提高5.03％和8.06％,料重比分别降低4.07％和5.81％,断奶仔猪的腹泻率分别降低74.90％和100.00％;试验Ⅰ组和Ⅱ组小猪的平均日增重较对照组分别提高1.60％和6.79％,料重比分别降低2.05％和5.13％.[结论]RLB可使断奶仔猪血清葡萄糖、IgA、白蛋白和总蛋白含量升高,血清胆固醇和IgG含量降低;RLB可以提高妊娠母猪、哺乳母猪、断奶仔猪和小猪的生产性能.     

传统发酵酸马奶中瑞士乳杆菌耐酸性的筛选

从新疆地区牧民家庭32份自然发酵酸马奶样品中分离鉴定出的82株瑞士乳杆菌中，通过耐酸实验选出39株可在pH3．0和pH3．5酸性环境中生长的菌株进行pH为3．0的人工胃液的耐受性实验。结果表明不同菌株对人工胃液的耐受能力有很大差异。     

干酪乳杆菌LC2W抗菌物质的研究

采用异丙醇抽提的方法，从干酪乳杆菌LC2W（Lb．casei CGMCC NO．0828）细胞中抽提到痢疾志贺氏菌（S．dysenteriae CMCC（B）51387）具有拮抗作用的蛋白类物质。该物质对蛋白酶尤其是胰蛋白酶敏感，对热不敏感。采用SDS—PAGE聚丙稀酰胺凝胶电泳的方法测得抑菌物质的分子量分别为32．1，44．3，87．6kDa。     

鸡源嗜酸乳扞菌的筛选

试验从健康雏鸡嗉囊、小肠、盲肠等部位分离到12株乳酸菌,通过试验筛选出了1株乳酸菌,初步鉴定为嗜酸乳杆菌,并对其耐酸、抑菌生物学特性进行初步研究。结果表明,嗜酸乳杆菌具有强耐酸性和黏附性,对大肠杆菌和金黄色葡萄球菌有很强的抑菌作用。     

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。     

青海部分牧区牦牛酸奶中乳杆菌的分离与鉴定

利用MRS培养基,对青海部分牧区收集的15份牦牛酸奶,进行了乳杆菌的分离,对分离得到的9株疑似乳杆菌,对其中的3株进行生物学鉴定。结果:分离株均为兼性厌氧菌,在MRS培养基上,菌落较大、透明、灰白色,镜检为杆状细菌,革兰氏染色阳性,过氧化氢酶试验为阴性,乳糖发酵阳性,牛乳发酵试验阳性,符合乳杆菌的特性。