培养基成分对植物乳杆菌LIP-1抗冷冻性的影响

以益生菌植物乳杆菌LIP-1为研究对象,MRS基础培养基为对照,通过改变培养基的成分及其含量（碳源、氮源、碳氮总量及碳氮比、缓冲盐）,研究培养基成分的变化对植物乳杆菌LIP-1高密度培养的活菌数以及冷冻干燥后存活率的影响。结果表明：通过优化工艺和配方确定的最佳培养基组成为碳源（蔗糖）添加量33.35 g/L、复合氮源（酵母粉、大豆蛋白胨、酪蛋白胨质量比3∶3∶5）添加量10.80 g/L、缓冲盐（乙酸钠、磷酸氢二钾、柠檬酸钠质量比2.5∶1∶1）添加量14.85 g/L（即0.13 mol/L）;用优化后     

保护剂对冷干和贮存过程中乳杆菌存活率的影响

研究以植物乳杆菌(Lactobacillus plantarum)为目标菌株,对冷冻干燥高活力乳酸菌粉的保护剂和保存条件进行了试验。结果表明:①海藻糖、谷氨酸钠、蔗糖、山梨醇和乳糖等5种保护剂对植物乳杆菌在冷冻干燥和干燥后贮存过程中均有显著的保护作用,其中,海藻糖的保护作用最为显著,植物乳杆菌存活率高达78.90%,而蔗糖、谷氨酸钠、乳糖和山梨醇作用稍差;②添加了不同保护剂制得的植物乳杆菌粉在4℃和25℃条件下的贮存结果表明,随着贮存温度的升高,植物乳杆菌的死亡率增加,随着保存时间的延长植物乳杆菌的死亡速度加快,添加海藻糖可以明显减缓植物乳杆菌的死亡速度。     

植物乳杆菌JM113的体外抗氧化能力及其对采食脱氧雪腐镰刀菌烯醇污染饲粮的肉仔鸡保护作用

本试验目的是研究从健康藏鸡肠道内容物分离出的植物乳杆菌JM113的抗氧化能力及其对脱氧雪腐镰刀菌烯醇污染的肉仔鸡保护作用。与植物乳杆菌PZ01和肠球菌M23相比,植物乳杆菌JM113在1.2 mmol/L的过氧化氢下表现出最强的还原性和抗性(P0.05),而且对羟基、超氧化物阴离子、1,1-二苯基-2-苦肼基自由基具有非常好的清除能力(P0.05)。对于每个菌株,活细菌菌株的抗氧化活性比不含细胞的提取物和死细菌菌株更强(P0.05)。为了检测植物乳杆菌JM113在体内抗氧化性能,试验选用192只1日龄爱拔益加肉鸡随机分为4组,每组6个重复,每个重复8只鸡,试验期42d。试验处理为(1)对照组;(2)对照组基础上添加1×109cfu/kg植物乳杆菌JM113;(3)对照组基础上添加10 mg/kg的脱氧雪腐镰刀菌烯醇;(4)在含10 mg/kg的脱氧雪腐镰刀菌烯醇的对照组基础上添加1×109cfu/kg植物乳杆菌JM113。与对照组相比,饲粮添加10 mg/kg的脱氧雪腐镰刀菌烯醇显著降低了试鸡血清中超氧化物歧化酶活性,并增加试鸡空肠粘膜中丙二醛含量(P0.05)。然而在对照组和脱氧雪腐镰刀菌烯醇污染组饲粮中添加植物乳杆菌JM113均显著降低了试鸡空肠粘膜中丙二醛含量(P0.05)。脱氧雪腐镰刀菌烯醇污染组试鸡空肠粘膜中核因子E2相关因子2(Nrf2)的表达水平显著降低(P0.05),而添加植物乳杆菌JM113处理后Nrf2及其相关的下游HO-1基因的转录水平显著升高(P0.05)。添加植物乳杆菌JM113可促进绒毛生长,甚至与脱氧雪腐镰刀菌烯醇联合在一起仍有显著效果(P0.05)。在植物乳杆菌JM113处理组,尤其是脱氧雪腐镰刀菌烯醇污染处理添加植物乳杆菌JM113可显著上调紧密连接蛋白claudin-1的表达(P0.05)。因此,本研究表明植物乳杆菌JM113菌株具有很强的抗氧化能力,添加进脱氧雪腐镰刀菌烯醇污染的饲料可保护肉仔鸡肠道屏障的完整性,减轻脱氧雪腐镰刀菌烯醇对家禽的负面影响。     

响应面法优化乳双歧杆菌Z-1冷冻干燥保护剂配方

采用响应面法对乳双歧杆菌Z-1冷冻干燥保护剂配方进行优化。通过Plackett-Burman试验与最陡爬坡试验确定影响乳双歧杆菌Z-1冷冻干燥后活菌数的显著因素及最优响应值区间，再通过Box-Benhnken试验优化得到乳双歧杆菌Z-1冷冻干燥保护剂最佳配方。结果表明：海藻糖、蔗糖、L-半胱氨酸盐酸盐添加量是影响乳双歧杆菌Z-1冷冻干燥后活菌数的显著因素；经响应面试验设计建立活菌数和海藻糖、蔗糖、L-半胱氨酸盐酸盐添加量的回归模型，得到乳双歧杆菌Z-1冷冻干燥保护剂最佳配方为海藻糖添加量200.51 g/L、蔗糖添加量46.16 g/L、L-半胱氨酸盐酸盐添加量2.31 g/L，其他成分不变（脱脂乳粉添加量100 g/L、乳清蛋白粉20 g/L、甘油4 g/L、异抗坏血酸钠10 g/L、谷氨酸钠8 g/L）。     

菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响

试验初步研究菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响。在MRS 培养基中添加1%、2%、3%和4%的菊粉,测定2株菌生长的OD600nm位和pH。结果表 明,当菊粉添加量为1%时,促进植物乳杆菌和保加利亚乳杆菌的生长,植物乳杆菌 生长OD600nm最高为2.799,pH从5.98降到3.59;保加利亚乳杆菌生长OD600nm最高 为3.051,pH从6.55降到4.74。     

腐植酸钠对植物乳杆菌生长、代谢及形态的影响

本试验通过在MRS液态培养基中添加不同浓度的腐植酸钠,比较植物乳杆菌生长、代谢及形态变化,初步探讨了液态条件下腐植酸钠对植物乳杆菌的抑制作用。结果表明,1%~5%腐植酸钠降低了植物乳杆菌的活菌数,但可提前6h达到生长最高点;3%腐植酸钠作用后,植物乳杆菌的有机酸产量发生了改变,且菌落变小,菌体变粗;透射电镜结果表明腐植酸钠使得植物乳杆菌细胞壁和细胞膜结构部分变得模糊。因此,液态条件下腐植酸钠通过改变植物乳杆菌的代谢途径、形态结构达到抑制其生长的作用。     

菊粉对植物乳杆菌和保加利亚乳杆菌生长的影晌

试验初步研究菊粉对植物乳杆菌和保加利亚乳杆菌生长的影响.在MRS培养基中添加1%、2%、3%和4%的菊粉,测定2株菌生长的OD600 nm位和pH.结果表明,当菊粉添加量为1%时,促进植物乳杆菌和保加利亚乳杆菌的生长,植物乳杆菌生长OD600nm最高为2.799,pH从5.98降到3.59;保加利亚乳杆菌生长OD600nm最高为3.051,pH从6.55降到4.74.     

不同乳酸菌制剂对金荞麦单贮及金荞麦和玉米淀粉混贮的影响

本研究旨在探讨不同乳酸菌制剂对金荞麦单独青贮,以及金荞麦与玉米淀粉混合青贮过程中的乳酸菌菌数、理化指标变化趋势及青贮效果的影响。以人工栽植的初花期金荞麦为青贮原料,选择3种复合乳酸菌制剂进行青贮。结果表明,金荞麦和玉米淀粉混合青贮的青贮品质明显优于金荞麦单贮,添加植物乳杆菌、布氏乳杆菌、屎肠球菌的菌粉组合对于金荞麦青贮品质的提升要显著优于植物乳杆菌、干酪乳杆菌菌粉组合和植物乳杆菌、发酵乳杆菌菌粉组合。综上,金荞麦不适宜单独青贮,需要配合一定比例的玉米淀粉进行混合青贮,植物乳杆菌、布氏乳杆菌和屎肠球菌的复合乳酸菌制剂更有利于金荞麦青贮品质的提升。     

自主筛选的植物乳杆菌对酸羊乳品质与功能特性的影响

为改善酸羊乳风味与品质,提高酸羊乳的功能价值,将自浆水中筛选的植物乳杆菌JS5和JS19用于羊乳发酵,研究其对酸羊乳理化性质、质构特性及功能特性的影响。结果表明：2 种植物乳杆菌与商业发酵剂复合发酵后,均能在贮藏期内有效提高酸羊乳持水性和表观黏度,降低其硬度与胶着性,促进乙醛和双乙酰的释放;添加植物乳杆菌JS5的酸羊乳胆固醇降解率显著高于对照组（P＜0.05）,最高可达36.62%;添加植物乳杆菌JS19的酸羊乳体外抗氧化能力更强,贮藏1 d时1,1-二苯基-2-三硝基苯肼自由基清除率高达70.58%,铁离子还原/抗氧化能力达32.03 μmol/L。2 种菌株均为适用于羊乳发酵生产的优良菌株。     

植物乳杆菌WEI-70菌粉对产蛋鸡的安全性及生产性能和蛋品质的影响

试验旨在研究基础日粮中添加植物乳杆菌(Lactobacillus Plantarum)WEI-70菌粉对产蛋鸡的生产性能、蛋品质、组织病理学影响,评价菌株WEI-70对产蛋鸡的安全性和功效性。试验使用305日龄海兰褐蛋鸡288只,随机分为2组,每组设6个重复,每个重复24只鸡,对照组饲喂基础日粮,试验组在基础日粮中添加植物乳杆菌WEI-70冻干菌粉,每克日粮中的活菌浓度为1×10~6 CFU。结果表明:与对照组相比,试验组的平均日产蛋量显著提高了2.71%(P0.05),平均料蛋比显著下降了3.27%(P0.05),哈氏单位显著提高了3.42%(P0.05),蛋黄颜色显著提高了4.37%(P0.05);试验组和对照组的心、肝、脾的脏器重量和脏器指数均无显著差异(P0.05),都没有出现病理学变化,但与对照组相比,试验组的空肠绒毛高度显著提高了31.82%(P0.05)。由此可见,植物乳杆菌WEI-70在蛋鸡养殖生产中能够促进蛋鸡生产性能、蛋品质的提升和空肠绒毛高度增加,并且不会引起脏器的病理变化,体现出较高的安全性。     

Collagen (Cn-I) binding by gut lactobacilli.

Four gut Lactobacillus strains displaying the features which make them particularly promising for the preparation of probiotic products were investigated together with 5 fresh isolates and one collection strain of Lactobacillus plantarum for their ability to bind type I collagen (Cn-I). Immobilised Cn-I in microtitre plates was bound only by 3 strains of gut lactobacilli from piglets and the collection strain Lactobacillus plantarum LHI 10 from Prague in range of A570nm readings 0.114-0.221. Six strains (isolates from turkeys and a calf) did not bind Cn-I (A570nm < 0.1) in this assay. An influence of cultivation medium on Cn-I binding was significant (P < 0.001) in all four adherent strains. Significantly higher (P < 0.001) binding of Cn-I was observed for Lactobacillus casei L 81 and Lactobacillus plantarum LHI 10 grown on solid medium (MRS agar) than for MRS broth-grown cells, however, Lactobacillus plantarum L 5 and Lactobacillus fermentum L 435 expressed significantly (P < 0.001) higher Cn-I binding during cultivation in MRS broth. The specificity of the binding was confirmed because the Cn-I binding by lactobacilli after their preincubation with this protein was completely abolished. Three selected inhibitors (fucoidan, heparan sulphate and hyaluronic acid) significantly (P < 0.001) reduced Cn-I binding by the Lactobacillus plantarum L 5 strain. Following up on some earlier strain characteristics, these results suggest that the selected piglets lactobacilli are also able to bind Cn-I and therefore should antagonize collagen niche colonization by various enteropathogens when used for probiotic purposes.     

猪源植物乳杆菌的诱变选育及其对小鼠免疫性能的影响

To get a Lactobacillus plantarum strain with good acid resistance, cholate tolerance, and in vitro adhesion rate and study its effects on immune function of mice. After screening Lactobacillus plantarum by acid resistance and cholate tolerance after UV mutagenesis, we got BL-15 strain, which was cultured, freeze-dried, dissolved and administered to BALB/c mice orally (gastric perfusion) by three times. Blood, spleen and excrement were collected to detect changes in immune function index.One Lactobacillus plantarum strain BL-15 was selected from 17 mutant strains, with higher acid resistance, cholate tolerance and in vitro adhesion rate. The experimental data of animal tests indicated that IL-2 level and spleen index of mice at early growth stage had been increased. Furthermore, the mice treated with BL-15 as the immunity enhancer revealed higher level of SIgA in intestinal contents compared with control (P<0.05). The results showed that Lactobacillus plantarum strain BL-15 had been proved to be able to elevate immune response and enhance immune function.     

植物乳杆菌的益生特性及其在乳制品中的应用

植物乳杆菌（Lactobacillusplanrarum）能通过胃并定植于肠道,发挥调节肠道菌群、免疫调节、降低血清胆固醇、降血压和抗氧化等益生作用。此外,由于植物乳杆菌的益生特性,国内学者申请了诸多关于植物乳杆菌菌株专利。同时,近年来有关植物乳杆菌分子多样性的研究也充分开展。另外,植物乳杆菌还被广泛应用于乳制品生产,包括功能性酸乳、发酵豆乳、干酪、功能性乳酸菌饮料等乳制品中。     

Survival and persistence of Lactobacillus plantarum 4.1 and Lactobacillus reuteri 3S7 in the gastrointestinal tract of pigs

Lactobacillus sp. are important inhabitants of the intestines of animals. They are also largely used as probiotics for both humans and animals. To exert beneficial effects, lactobacilli have to survive through the gastrointestinal transit. Based on bile-salt resistance, pH tolerance, antimicrobial activity and heat resistance, Lactobacillus plantarum 4.1 and Lactobacillus reuteri 3S7 were previously selected and used as probiotic additives in pelleted feeding trials. Both strains were fed to pigs (sows and piglets) at a cell number of ca. 10(10) viable cells per day. A polyphasic approach, comprising growth on selective media, Biolog System analysis, 16S rRNA gene sequencing and RAPD-PCR typing, was used to identify and differentiate L. plantarum 4.1 and L. reuteri 3S7 from other faecal Lactobacillus sp., L. plantarum 4.1 and L. reuteri 3S7 had the capacity to survive during the gastrointestinal transit and were found in the feaces at a cell number of 6-8 log cfu/g. Their persistence was shown after 6 days from the administration. Compared to untreated pigs, the administration of L. plantarum 4.1 and L. reuteri 3S7 significantly (P<0.05) decreased the population of Enterobacteriaceae. Besides, the beta-glucuronidase activity of all pigs decreased of ca. 23.0% during administration. The findings of this study showed that L. plantarum 4.1 and L. reuteri 3S7 have the potential to be used as probiotic additives in pelleted feed for pigs.     

猪源乳酸杆菌的安全性及抗逆性评价

对本实验室自行分离自猪肠道的3株乳酸杆菌进行安全性和抗逆性检测,采用体外法对分离自不同生理阶段猪(仔猪、生长肥育猪、母猪)肠黏膜和食糜的罗伊乳杆菌(Lactobacillus reuteri)LA5、植物乳杆菌(L.plantarum)L47和约氏乳杆菌(Ljohnsonii)L63进行生长特性、溶血性、有害代谢产物、...     

Lectin-like binding of lactobacilli considered for their use in probiotical preparations for animal use

Three gut lactobacilli from piglets (Lactobacillus plantarum L 5, Lactobacillus paracasei L 81, Lactobacillus fermentum L 670) and Lactobacillus casei subsp. pseudoplantarum L.c.) from a calf were examined by microtitre plate binding assay for their lectin-like binding activity after their cultivation on Rogosa agar and in MRS broth. Three ECM (extracellular matrix) molecules (fetuin, porcine fibronectin and porcine mucin) were selected for this assay. Additionally, the effect of heparin on the binding of these three ECM molecules by Lactobacillus strains in microtitre plates was tested. Moreover, haemagglutination tests with pig, cattle, sheep, and hen erythrocytes were performed. However, none of the four Lactobacillus strains examined did react with any of the erythrocytes tested. The differences between individual strains were observed in their binding to immobilised ECM molecules. The best adherent was the Lactobacillus plantarum L5, however, the other three strains showed also good ECM binding. With regard to an influence of cultivation medium on lectin-like binding activity, binding of all ECM molecules was expressed in Lactobacillus paracasei L 81 to significantly higher degree (P < 0.001) after cultivation on Rogosa agar than in MRS broth. Similarly, strains Lactobacillus fermentum L 670 and Lactobacillus casei subsp. pseudoplantarum L.c. displayed significantly higher (P < 0.001) binding of fibronectin and mucin after growth on Rogosa agar in comparison with MRS broth cultivation. However, no significant (on fetuin and fibronectin binding) or opposite effect (on mucin binding) of cultivation medium was observed in Lactobacillus plantarum L 5 strain. The influence of cultivation medium on fetuin binding by Lactobacillus fermentum L 670 was also not significant while Lactobacillus casei subsp. pseudoplantarum L.c. bound fetuin significantly better (P < 0.01) after growth on Rogosa agar. Heparin pretreatment increased the binding of the ECM molecules by the Lactobacillus fermentum L 670 strain significantly (P < 0.001 or P < 0.05) with the exception of porcine fibronectin when the strain was cultivated in MRS broth. This result is important especially in the connection with the previous observations that heparin decreased ECM binding of enteropathogens as staphylococci or clinical enterococcal isolates. Following up on some earlier strain characteristics, these results indicate that the selected lactobacilli are probably suitable for probiotic purposes.     

植物乳酸杆菌对感染沙门氏菌蛋鸡血浆生化、免疫水平及沙门氏菌定植的影响

本试验旨在研究植物乳酸杆菌对感染肠炎沙门氏菌蛋鸡血浆生化、免疫水平及沙门氏菌定植的影响。将400只京红1号商品代蛋鸡随机分成4个处理组,每组5个重复,每个重复20只鸡。A组和C组饲喂基础饲粮,B组和D组在基础饲粮添加2×10~8 CFU/g植物乳酸杆菌。饲喂1周后,C组和D组连续2 d口服1×10~10CFU/m L肠炎沙门氏菌菌液,A组和B组口服等量无菌PBS溶液,饲养3周。结果表明:添加植物乳酸杆菌可显著提升总蛋白、球蛋白水平(P0.05),显著降低低密度脂蛋白和甘油三酯(P0.05);添加植物乳酸杆菌可显著增加IgM、IgG(P0.05);蛋鸡感染沙门氏菌的第14天和第21天及试验全程,添加植物乳酸杆菌显著减少盲肠食糜中肠炎沙门氏菌拷贝数(P0.05)。综上可知,日粮中添加植物乳酸杆菌可改善感染肠炎沙门氏菌蛋鸡的血浆生化指标,缓解炎症损伤,降低盲肠沙门氏菌定植数量,保障机体健康。     

植物乳酸杆菌对感染肠炎沙门氏菌蛋鸡生产性能、蛋品质及血浆生化指标的影响

为研究植物乳酸杆菌对感染肠炎沙门氏菌蛋鸡生产性能、蛋品质及血浆生化指标的影响,本试验选用400只沙门氏菌阴性蛋鸡,随机分成4个处理组,每处理组5个重复,每个重复20只试鸡。试验第1周,对照组（T1、T3组）饲喂基础日粮,试验组（T2、T4组）在基础日粮添加2×10⁸ CFU/g植物乳酸杆菌。饲喂1周后,T3、T4组试鸡连续2 d灌服1 mL肠炎沙门氏菌悬浮液（1.0×10⁸ CFU/mL）,T1、T2组试验鸡口服等量无菌PBS溶液。结果显示：①蛋鸡感染肠炎沙门氏菌后可极显著降低产蛋率（P_SE=0.02）及平均日采食量（P_SE=0.01）。添加植物乳酸杆菌可显著提高产蛋率（P_LP=0.02）,显著降低料蛋比（P_LP=0.04）。②蛋鸡感染肠炎沙门氏菌后,添加植物乳酸杆菌可显著增加蛋壳厚度（P_LP=0.01）和蛋壳强度（P_LP=0.02）,显著降低蛋黄颜色（P_LP=0.02）。③蛋鸡感染肠炎沙门氏菌后可极显著提高蛋鸡血浆中碱性磷酸酶（ALP）含量（P_SE<0.01）,植物乳酸杆菌干预可显著提高谷草转氨酶（AST）（P_LP=0.02）、钙（P_LP=0.04）和磷（P_LP=0.04）的含量。综上所述,植物乳酸杆菌可显著改善蛋鸡生产性能和蛋品质,维持蛋鸡生理生化指标稳定,同时可缓解沙门氏菌带来的病理损伤,保障蛋鸡机体健康。     

植物乳杆菌、布氏乳杆菌对甘蔗尾青贮品质及有氧稳定性的影响

为了研究植物乳杆菌、布氏乳杆菌对甘蔗尾青贮品质及有氧稳定性的影响。试验共分为10组,A组为对照组,添加生理盐水;B、C、D组为植物乳杆菌添加组,分别在新鲜甘蔗尾中添加10、20和30 mL/kg植物乳杆菌液;E、F、G组为布氏乳杆菌添加组,在新鲜甘蔗尾中分别添加10、20和30 mL/kg布氏乳杆菌液;H、I、J组为植物乳杆菌、布氏乳杆菌联合添加组,在新鲜甘蔗尾中分别添加10、20和30 mL/kg布氏乳杆菌和植物乳杆菌等体积混合的菌液。每组3个重复。室温青贮40 d,结束后采样测定青贮发酵品质和有氧稳定性。结果显示：与对照组相比,添加植物乳杆菌组降低了甘蔗尾青贮pH,显著增加乳酸含量（P<0.05）,且随着植物乳杆菌添加量的升高而升高。与添加植物乳杆菌相比,添加布氏乳杆菌组甘蔗尾青贮中乳酸含量显著降低（P<0.05）,乙酸含量显著升高（P<0.05）,且均随着布氏乳杆菌添加量的提高效果更显著;添加布氏乳杆菌组甘蔗尾青贮有氧稳定性较对照组提高48 h。布氏乳杆菌和植物乳杆菌联合组甘蔗尾青贮中乳酸含量显著高于对照组（P<0.05）,且干物质损失较单独添加布氏乳杆菌组有所降低,且能够提高青贮可溶性碳水化合物（WSC）利用效率。综上所述,植物乳杆菌与布氏乳杆菌联合处理（添加量达到2×10⁶ CFU/g鲜重,I组）青贮甘蔗尾能够有效提高青贮品质和有氧稳定性。