唾液乳杆菌的生长特性及抑菌作用

为了探讨唾液乳杆菌的生长特性及抑菌作用,测定2株唾液乳杆菌的生长曲线,并选用口腔变形链球菌、牙龈卟啉单胞菌及金黄色葡萄球菌为指示菌,采用牛津杯法进行体外抑菌实验.结果表明:2株唾液乳杆菌分别在接种2h和4h进入对数生长期,8h进入对数生长末期,14h和16h进入稳定期;2株唾液乳杆菌菌液及代谢产物对口腔变形链球菌、牙龈卟啉单胞菌及金黄色葡萄球菌均有抑菌作用,对口腔变形链球菌及牙龈卟啉单胞菌的抑菌作用强于金黄色葡萄球菌;2株唾液乳杆菌菌体及调整pH值为7.0的代谢产物没有抑菌作用;2株唾液乳杆菌的抑菌能力主要来自菌体的代谢产物,代谢产物中起抑菌作用的成分为酸或酸依赖性物质.     

乳酸杆菌抑制致病菌及生物保鲜发酵剂特性评价

以15株乳酸杆菌为研究对象,动物双歧杆菌乳亚种BB-12作为对照,研究它们对肠道致病菌的抑菌特性.结果表明:9株菌对5株指示菌均有较好抑制作用,2株菌对金黄色葡萄球菌、肠炎沙门氏菌、大肠杆菌和蜡样芽孢杆菌均有抑制作用;以抑菌效果、凝乳状态、产酸性能为指标,筛选出4株菌,与嗜热链球菌ST-21组合制备生物保鲜发酵剂;通过...     

鸡源耐镉乳酸菌株的分离与鉴定

本试验旨在从鸡肠道中分离筛选出耐镉(Cd)的乳酸菌,为将其开发应用于防控家禽饲料重金属Cd污染提供理论基础。取鸡肠道内容物的样品进行乳酸菌分离,以耐受100 mg/L的Cd为标准筛选乳酸菌,采用分子生物学方法鉴定菌株。对筛选出的乳酸菌株进行生长性能、耐Cd性能及Cd吸附性能等测定。结果表明：本试验共筛选出5株耐Cd乳酸菌,分别为1株约氏乳杆菌Cd3-1和4株卷曲乳杆菌Cd1-1、Cd5-2、Cd4-5和Cd8-2。其中Cd3-1、Cd5-2和Cd8-2菌株最高可耐受1 000 mg/L的Cd,而Cd1-1和Cd4-5菌株最高可耐受400 mg/L的Cd。各菌株对50 mg/L Cd的吸附率随溶液pH的升高而增加,在pH为6.0和7.0时Cd的吸附率均显著高于pH为2.6时Cd的吸附率(P<0.05)。5株乳酸菌均可耐受0.3%的胆盐,且可抑制金黄色葡萄球菌的生长。综合上述结果,本试验从鸡肠道中分离筛选鉴定出5株耐Cd乳酸菌,包括1株约氏乳杆菌和4株卷曲乳杆菌,在pH接近中性的条件下,对Cd的吸附率可达30%。     

脱脂乳发酵体系中乳酸菌的高效离子交换色谱分析

应用高效离子交换色谱对脱脂乳发酵体系中的嗜热链球菌和保加利亚乳杆菌进行色谱表征。结果表明，单菌培养过程中保加利亚乳杆菌菌体形态的变化在其相应色谱行为上得到了体现，二者的色谱峰有比较好的稳定性。两种菌的峰面积与菌数之间呈现出良好的线性关系，可决系数R^2分别为0．9957和0．9916。因此本方法的建立，可用于酸奶发酵过程中乳酸菌的分析与表征，有望为乳酸菌混合物的研究提供一种快捷的新方法。     

基于PFGE技术的酸乳中乳酸菌的基因分型

采用选择性培养基分离酸乳中的保加利亚乳杆菌和嗜热链球菌,并利用生理生化和糖发酵实验结合16S rDNA同源性分析对分离所得菌株进行鉴定,获得保加利亚乳杆菌和嗜热链球菌分离菌株。利用脉冲场凝胶电泳（pulsed field gel electrophoresis,PFGE）分别对保加利亚乳杆菌和嗜热链球菌分离株进行基因分型。结果表明：所检测的酸乳样品中均含有保加利亚乳杆菌和嗜热链球菌;仅有2 种酸乳的嗜热链球菌为同一菌株,其余酸乳中的保加利亚乳杆菌及嗜热链球菌均为不同菌株。PFGE可以对保加利亚乳杆菌和嗜热     

In vitro growth inhibition of major mastitis pathogens by Staphylococcus chromogenes originating from teat apices of dairy heifers

Earlier field observations suggest that teat apex colonization by Staphylococcus chromogenes pre-partum in dairy heifers protects udder quarters against elevated somatic cell counts early post-partum. To explain these findings, the in vitro inhibitory capability of S. chromogenes from teat apices of heifers towards some major mastitis pathogens was tested using a modified cross-streaking method. Two out of 10 S. chromogenes isolates, both originating from two different teats from the same heifer, consistently inhibited growth of all Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis strains, but none of the Escherichia coli strains. The present study, therefore, supports the protective effect of teat apex colonization by S. chromogenes by in vitro production of inhibitory substances.     

不同来源乳酸菌的分离与鉴定

Six strains of Lactobacillus were isolated from traditional indigenous dairy products, piglet’s feces and chicken intestine, designated as LS, LT, LJ1, LJ2, LZ1 and LZ2， respectively. Identifications were done according to bacteria morphology, physiological and biochemical properties,16S rDNA sequence homology analysis, and their characteristics were researched. The results showed that LS, LT, LJ1 and LZ2 strains were Lactobacillus brevis, LJ2 strain was Enterococcus faecium, LZ1 strain was Lactobacillus acidophilus. All of the six strains had obvious bacteriostasis effect on Listeria monocytogenes, Salmonella typhimurium, Escherichia coli and Staphylococcus aureus.     

德氏乳杆菌保加利亚亚种和嗜热链球菌的共生机制研究进展

在发酵乳制品中,德氏乳杆菌保加利亚亚种和嗜热链球菌的共生对于发酵的影响至关重要,并对各自环境的适应尤其是酸环境的适应尤为重要.本文通过双组分信号转导系统、基因水平转移、蛋白质代谢、核苷酸碱基代谢、糖代谢、氧化性应激等方面对二者共生机制进行讨论,从而为研究和提高发酵制品的品质提供借鉴.     

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。     

嗜热链球菌对瑞士乳杆菌发酵乳后酸化的影响

瑞士乳杆菌是潜在的功能发酵乳发酵剂,为改善瑞士乳杆菌的发酵特性,使用嗜热链球菌配合瑞士乳杆菌用于发酵乳制备。结果表明：嗜热链球菌与瑞士乳杆菌混合发酵将凝乳时间缩短了3.5 h,黏度提高了1.8 倍,改善了乳清析出和质构;嗜热链球菌的使用有效减弱了发酵乳的后酸化,贮藏20 d发酵乳的pH值仍大于4.2。为研究这一现象的原因,测定发酵乳中的活菌数、乳糖消耗量、谷氨酸脱羧酶和β-半乳糖苷酶活性,结果表明,混合发酵乳贮藏期间具有较低的谷氨酸脱羧酶和β-半乳糖苷酶活性,乳糖利用量较低,这是后酸化减弱的主要原因。与嗜     

Isolation of surface (S) layer protein carrying Lactobacillus species from porcine intestine and faeces and characterization of their adhesion properties to different host tissues

Surface-layer proteins (Slps) of lactobacilli have been shown to confer tissue adherence. This study aimed to isolate and identify Slps carrying Lactobacillus species from the porcine intestine and faeces and to characterize these S-layer-expressing strains for their ability to adhere to the pig and human intestinal cells and to extracellular matrix (ECM) proteins. In total 99 strains, putatively belonging to the genus Lactobacillus, were isolated as pure cultures. SDS-PAGE and a gene probe specific for the Lactobacillus brevis ATCC 8287 S-layer protein gene (slpA) were used to screen the presence of strains possessing putative Slps. Eight of the 99 pure cultures exhibited Slps according to the SDS-PAGE analyses. In these strains the presence of genes encoding Slps was confirmed by PCR and partial sequencing. Only one isolate of the 99 strains gave a positive hybridisation signal with the L. brevis slpA probe but did not appear to produce S-layer protein. Their taxonomic identification, based on phenotyping and the 16S rRNA sequences, revealed that the eight S-layer protein-producing strains were closely related to Lactobacillus amylovorus, Lactobacillus sobrius and Lactobacillus crispatus. The strain with the slpA positive hybridisation result was identified as Lactobacillus mucosae. The SDS-extractable protein profile, the size of the putative S-layer protein and binding capability of the strains varied greatly, even among the isolates belonging to the same Lactobacillus cluster. Removal of the intact Slps from the bacterial surface by extraction with guanidine hydrochloride reduced the adhesion of some strains to fibronectin and laminin, whereas, the adhesiveness to laminin increased with some strains.     

干酪生产用菌种的筛选

采用从新疆民族乳制品中分离的菌株，研究了三种单一菌株（L1乳酸乳球菌、L2瑞士乳杆菌、L3干酪乳杆菌）和四种混合菌株（L4乳酸乳球菌：瑞士乳杆菌（1：1）、L5乳酸乳球菌：干酪乳杆菌（1：1）、L6瑞士乳杆菌：干酪乳杆菌（1：1）、L7嗜热链球菌：保加利亚乳杆菌（1：1））的发酵性能和生化特性，确定了适于干酪加工的菌种为L1、L5、L6。     

健康鸡嗉囊和盲肠内容物正常菌群的分离与鉴定

用血琼脂平板和乳清琼脂平板，采用集性没食子酸厌氧培养法，从健康非免疫产蛋母鸡的嗉囊和盲肠内容物中分离出正常菌群，地其中２１株瘘性厌氧的无芽胞杆菌和球菌，从革兰氏染色特性、形态特性、需氧生长条件和生化反应等方面进行了初步鉴定。结果其中有类肠膜明串珠菌１株，肠膜明串珠菌１株，粪链球菌３株，屎锭球菌１株，鸡肠链球菌１少酸链力３株，乳链球菌１株，假长双歧杆菌１株，星状双歧杆菌１株，嗜酸乳杆菌１株，嗜粪乳杆     

发酵乳后熟期间嗜热链球菌和保加利亚乳杆菌产乙醛特性

目的:分析嗜热链球菌和保加利亚乳杆菌在发酵乳后熟期间产乙醛特性,并研究乙醛合成与其关键调控基因表达量之间的关系.方法:以传统发酵乳制品中筛选出的具有优良发酵特性的嗜热链球菌和保加利亚乳杆菌为研究对象,将各试验菌株在复原全脂乳中进行单菌发酵,发酵结束后(pH4.5～4.6)于4℃冷藏后熟,测定48 h内发酵乳中的乙醛含量;采用反转录定量PCR技术检测乙醛合成关键调控基因pdc、pdh、ald、ldh的表达特征.结果:6株嗜热链球菌产乙醛量介于2.59～14.53 μg/g之间,6株保加利亚乳杆菌产乙醛量介于9.17～39.45 μg/g之间;乙醛合成量随着基因pdc、ald及pdh表达量的升高而增加,而与基因ldh的表达量呈负相关.结论:发酵乳后熟期间嗜热链球菌和保加利亚乳杆菌不同菌种、不同菌株乙醛产量差异显著,相同菌株在不同后熟时间产乙醛量差异明显,存在菌株特异性和时序变异性;调控基因pdc、pdh及ald具有促进乙醛合成的作用,基因ldh的表达不利于乙醛含量的积累.     

可利用表乳糖的仔猪肠道乳杆菌筛选及体外抑菌活性评价

本研究通过采用高通量测序、平板筛选、16S rDNA序列分析、特异性碳源培养和牛津杯法,从哺乳期健康仔猪的粪便中筛选分离和鉴定乳杆菌,并进行其利用表乳糖生长特性以及抑制仔猪腹泻相关致病菌活性的试验研究,探索表乳糖定向调控仔猪肠道乳杆菌增殖的可能性。结果表明:从7~10日龄哺乳期健康仔猪的粪便样品中分离出9株乳杆菌,其中唾液乳杆菌、卷曲乳杆菌、淀粉乳杆菌具有较强利用表乳糖的能力,在以2%表乳糖为唯一碳源的MRS培养基中显现出良好的生长态势,而且其培养上清液对大肠杆菌(猪源)、猪霍乱沙门氏菌、单核细胞增生李斯特氏菌、宋氏志贺氏菌以及金黄色葡萄球菌具有较好的抑菌活性。     

4株Lactobacillus菌株对结肠癌细胞增生的抑制效果研究

以分离自传统发酵乳制品中的4株菌——Lactobacillus paracasei supsp.paracasei M5AL、L.paracasei supsp.paracasei J23ANL、L.paracasei supsp.paracasei G15AL、L.coryniformis supsp.torquens T3AL为实验菌株,采用噻唑兰法研究其热灭活菌体、细胞壁及DNA对结肠癌细胞HT-29增殖的影响。结果表明：菌株J23ANL、M5AL、G15AL、T3AL热灭活菌体细胞、菌体细胞壁和菌体DNA均可不同程度地抑制人结肠癌细胞HT-29的增生,其中菌株T3AL对HT-29细胞增生的抑制率显著高于其他菌株的抑制率（P〈0.05）;单细胞凝胶电泳结果显示菌株T3AL可诱导HT-29细胞凋亡。     

散养成年蛋鸡空肠乳酸菌群的区系分析

试验以5只55周龄散养健康成年蛋鸡空肠内容物为样本,用无菌生理盐水稀释至10-6稀释浓度,吸取0.1 mL菌液滴于MRS平板上,置37℃培养箱培养24 h,分离乳酸菌。将分离纯化得到的乳酸菌,通过革兰氏染色、菌落形态观察、糖发酵试验进行乳酸菌种属鉴定。结果显示,从55周龄散养健康成年蛋鸡空肠中分离筛选出20株乳酸菌,经革兰氏染色后镜检显示,分离菌株有杆状和球形2种;经糖发酵试验鉴定为7个乳酸菌种,分别为唾液乳杆菌唾液亚种、棒状乳杆菌扭曲亚种、唾液乳杆菌水杨素亚种、能动乳杆菌、格氏乳杆菌、嗜酸乳杆菌和德氏乳杆菌保加利亚亚种。     

抑霉乳酸菌的筛分及其抑菌特性研究

试验旨在获取生物抑霉的优良菌株以及为质量安全青贮饲料生产提供高效菌剂。从青贮饲料、酸奶以及奶豆腐中分离纯化出乳酸菌和霉菌,并且通过双层平板法对具有抑制霉菌活性的乳酸菌菌株进行筛选,测定其上清液抑霉效果;采用蛋白酶处理和高温处理法分析乳酸菌抑霉的有效成分。结果表明:分离筛选的乳酸菌和霉菌经鉴定为植物乳杆菌和黄曲霉菌,且植物乳杆菌对黄曲霉菌的生长有明显抑制作用。蛋白酶处理对乳酸菌无细胞上清液的抑菌活性有不同影响,而加热处理并不改变其抑菌效果。研究表明,植物乳杆菌对黄曲霉菌生长有明显的抑制作用,推测其抑菌活性物质为蛋白质、肽类等,并且这些抑菌活性物质的热稳定性较高。     

不同发酵剂对褐色乳酸菌饮料的影响

[目的]发酵剂是影响褐色乳酸菌饮料品质的关键原料之一。本文测试不同型号的发酵剂对褐色乳酸菌饮料发酵时间、产品风味、稳定性的影响。[方法]以市面上最常用的褐色乳酸菌饮料发酵剂之一副干酪乳杆菌L.casei-01作为参照,另外购买2 种副干酪乳杆菌作为对比,进行测试。[结果]本文测试的发酵剂中,参照样副干酪乳杆菌L.casei-01发酵速度最慢,但风味层次感较强。副干酪乳杆菌L9s相较于参照样,发酵速度更快,但风味偏酸。副干酪乳杆菌Lpc-37相较于参照样,发酵速度更快,但稍慢于L9s,风味较柔和。3 种发酵剂对产品的离心沉淀率、动力学不稳定性影响较小。[结论]3 种发酵剂对产品发酵速度、风味影响较大;对产品的稳定性影响较小。     

Putative probiotic Lactobacillus spp. from porcine gastrointestinal tract inhibit transmissible gastroenteritis coronavirus and enteric bacterial pathogens

A total of 310 bacterial strains isolated from the porcine gastrointestinal tract were tested for their activity against transmissible gastroenteritis (TGE) coronavirus and other enteric pathogens. Based on activity, the strains Probio-38 and Probio-37 were selected as potential probiotics and identified as Lactobacillus plantarum Probio-38 and Lactobacillus salivarius Probio-37 respectively by 16S rRNA gene sequencing. Supernatants of these strains inhibited TGE coronavirus in vitro in ST cells, without any cytopathic effect even after 72 h of incubation. Both the strains exhibited high survival in synthetic gastric juice. The strains were resistant to 5% porcine bile and exhibited antimicrobial activity against all the 13 enteric bacterial pathogens tested. These strains also exhibited resistance to most of the antibiotics analyzed. The inhibition of transmissible gastroenteritis coronavirus and enteric bacterial pathogens as well as the bile tolerance, high survival in gastric juice, and the antibiotic resistance indicate that the two isolated bacterial strains are ideal probiotic candidates for animal application after proper in vivo experiments.