热压结合处理对绿色魏斯氏菌生理特性的影响

为了探究热压结合处理在绿色魏斯氏菌致死过程中发挥的作用。本实验采用超高压(350 MPa、25℃、10 min)和热压结合(350 MPa、50℃、10 min)对绿色魏斯氏菌进行压力处理,研究了热压结合处理对绿色魏斯氏菌生理特性、细胞形态特征、细胞膜的通透性等的影响。结果显示,热压结合处理使菌落总数从超高压处理时的7.67(lg(CFU/mL))显著下降到5.71(lg(CFU/mL))(P0.05),协同的热处理使菌体表面出现严重褶皱、菌体内含物变性凝固加剧,使细胞膜损伤,从而改变膜的通透性,导致细胞外紫外吸收物质显著增加(P0.05)。因此,热压结合处理在超高压致死细菌中会加剧细胞膜的形变及损坏,增加细胞膜的通透性,从而导致细胞内紫外吸收物质的流出,实验结果为热压结合灭活绿色魏斯氏菌提供了理论依据。     

DMDC联合Nisin对模拟果汁中肠膜状明串珠菌细胞膜功能的影响

本文研究了二甲基二碳酸盐(DMDC)和Nisin处理对模拟果汁中肠膜状明串珠菌的杀菌效果及其细胞膜功能的影响。结果表明:DMDC联合Nisin作用于该菌时的部分杀菌浓度指数为0.38,小于0.50,两者之间具有很好的协同杀菌作用。扫描电镜分析发现该菌经DMDC和Nisin处理后其细胞形态没有发生明显变化。该菌经DMDC处理后仅有个别菌体细胞的膜通透性出现增加,而该菌经Nisin处理后,约7%菌体细胞的膜通透性增加。Nisin处理虽能改变该菌的细胞膜通透性,增加胞外极性物质的摄入,但并不能明显促进胞内物质的流失,高浓度的DMDC处理能导致该菌溶液的紫外吸收值增加约60%。DMDC和Nisin两者对该菌细胞内物质的流失、细胞膜的通透性的增加和胞内p H的下降没有相互促进作用,但DMDC和Nisin的联合作用能促进该菌细胞内脱氢酶的进一步失活。     

超高压协同乳酸链球菌素抑制低温火腿中的耐压腐败菌

为探寻低温肉制品中耐压腐败菌的抑制手段,揭示超高压和乳酸链球菌素之间的协同抑菌效应。以肉制品中典型的耐压腐败菌绿色魏斯菌和肠膜明串珠菌为供试材料,评价二者在1～500μg/mL的乳酸链球菌素存在条件下的生长情况,并结合100～600MPa(5min,20℃)的超高压处理研究不同压力条件与乳酸链球菌素之间的协同增效作用。结果表明:随着乳酸链球菌素添加量的增加,两种菌的数量急剧降低,当添加量达到200μg/mL时,对二者的抑制程度达到最大;单独超高压处理时,绿色魏斯菌和肠膜明串珠菌分别在400MPa和300MPa的压力下受到显著抑制。而超高压结合200μg/mL乳酸链球菌素处理时,二者受到显著抑制的压力均降低到100MPa;500MPa,5min,20℃的超高压条件结合200μg/mL乳酸链球菌素处理能够抑制绿色魏斯菌数达9lg(cfu/mL)。超高压和乳酸链球菌素之间存在显著的协同作用,乳酸链球菌素的添加将会有效降低工业化生产中超高压的压力水平。     

乳制品中明串珠菌的分离鉴定及其生物学特性研究

对内蒙古自治区呼伦贝尔市牧区98个乳样中的明串珠菌进行了分离及生物学特性研究。共分离到明串珠菌3株，分别属于乳明串珠菌(EW21)，肠膜明串珠菌葡聚糖亚种(EW24—2)和肠膜明串珠菌肠膜亚种(CB8—1)。它们均可在pH=4．8的环境中生长，且菌株EW24—2和CB8—1可利用蔗糖生成葡聚糖。     

假肠膜明串珠菌产细菌素复配抑菌液的筛选及保鲜效果研究

目的 筛选出最优的假肠膜明串珠菌（Leuconostoc mesenteroides, LM）产细菌素复配抑菌液,同时探究抑菌液对冷鲜鸡肉的保鲜效果。方法 以冷鲜鸡肉及其主要腐败菌为研究对象,研究假肠膜明串珠菌产细菌素的最佳生长周期,并利用假肠膜明串珠菌产细菌素抑菌液、乳酸链球菌素抑菌液、假肠膜明串珠菌产细菌素和柚子精油复配抑菌抑菌液以及假肠膜明串珠菌产细菌素和茶多酚复配抑菌液分别浸泡鸡肉,从肉的pH、汁液流失率、色度、菌落总数、挥发性盐基氮（total volatile basic nitrogen, TVB-N）、感官评定等评价其对鸡肉冷藏品质的影响,比较以上几种抑菌液对鸡肉的保鲜效果,筛选出最佳的抑菌液。结果 假肠膜明串珠菌产细菌素的生长周期与其抑菌活性呈一定正相关。实验组的4种抑菌液均能延缓冷鲜鸡肉劣变,但1.5%假肠膜明串珠菌产细菌素抑菌液、40 mg/L乳酸链球菌素抑菌液对冷鲜鸡肉保鲜效果有限。而1.5%假肠膜明串珠菌产细菌素和0.2%茶多酚复配抑菌液、1.5%假肠膜明串珠菌产细菌素和10%柚子精油复配抑菌液的保鲜效果较好,尤其在以1.5%假肠膜明串珠菌产细菌素和10%柚子精油复配抑菌液液处理时,对冷鲜鸡肉的保鲜作用最强。结论 1.5%假肠膜明串珠菌产细菌素和10%柚子精油复配抑菌液对冷鲜鸡肉具有最佳的保鲜效果。     

发酵香肠中分离纯化的三株乳酸菌产酸特性研究

为了研究从发酵香肠中分离纯化的3株乳酸菌粪肠球菌(Enterococcus faecalis)、戊糖片球菌(Pediococcus pentosaceus)和肠膜明串珠菌(Leuconostoc mesenteroides)的产酸性能,对这3株菌在不同pH、温度、NaCl、NaNO2条件下的生长情况及产酸情况进行了测定。研究结果显示:这3株菌中,肠膜明串珠菌和戊糖片球菌的生长特性较好;粪肠球菌的生长特性虽不如肠膜明串珠菌和戊糖片球菌,但产酸能力最好,戊糖片球菌耐盐性最好、肠膜明串珠菌耐亚硝酸盐的特性最好;在不同温度和pH条件的测试中,肠膜明串珠菌的生长能力最好,粪肠球菌次之。这3株乳酸菌在发酵肉制品中均产生乳酸。总之,这3株菌均具有用于发酵制备乳酸的能力。     

肠膜明串珠菌的研究和应用进展

肠膜明串珠菌是乳酸菌中的明串珠菌属的重要菌种。研究证实,肠膜明串珠菌能发酵糖类产生多种酸和醇,具有高产酸能力、抗氧化能力和拮抗致病菌等能力。目前肠膜明串珠菌已被广泛应用于风味剂、血浆代用品,还有望成为新型微生态制剂。     

食窦魏斯氏菌NCU034005与柠檬明串珠菌NCU027003对泡菜代谢物及风味的影响

在灭菌后的泡菜原材料中分别接种食窦魏斯氏菌NCU034005与柠檬明串珠菌NCU027003,室温纯种发酵7d,期间检测泡菜液中挥发性物质与非挥发性物质(葡萄糖、有机酸、氨基酸等)、pH、风味物质香气活度值。结果表明:食窦魏斯氏菌NCU034005与柠檬明串珠菌NCU027003纯种发酵泡菜中pH不同程度下降,两株乳酸菌对碳水化合物、有机酸与氨基酸的代谢能力不同;主成分分析表明食窦魏斯氏菌NCU034005与柠檬明串珠菌NCU027003纯种发酵泡菜中的代谢物与风味存在显著差异,且有自身的代表性标志代谢物与风味因子。食窦魏斯氏菌NCU034005与柠檬明串珠菌NCU027003对泡菜代谢物与风味有显著影响。     

SDS-PAGE电泳法分析朝鲜族辣白菜中乳酸菌分布

为了分析辣白菜发酵过程中乳酸菌分布,从自然发酵的自制朝鲜族辣白菜中运用平板稀释法根据细菌菌落形态特征分离筛选出59株乳酸菌,SDS-PAGE电泳法分析各菌株全细胞蛋白模式后将其划分为7大类,最后用16S rRNA序列同源性分析,鉴定出4种乳酸菌种类,分别为肠膜明串珠菌、柠檬明串珠菌、食窦魏斯氏菌和植物乳杆菌。     

碳酸饮料絮凝物的微生物分离及其耐酸性研究

从碳酸饮料的絮凝物中分离出的微生物,经过形态学特征及生理生化特性分析,获得了3株明串珠菌属菌株,鉴定为:假肠膜明串珠菌1株,肠膜明串珠菌葡聚糖亚种2株。对3株菌株进行了pH3.0的耐酸性研究,其中3#呈现出最强的耐酸性和碳酸饮料的适应性。     

真空包装熟肉制品中的特定腐败微生物及其控制

真空包装低温熟肉制品的腐败变质主要是由乳酸菌引起的,腐败突出表现在出汁、产酸、产气、发粘4种感官腐败.主导腐败的微生物由清酒乳杆菌、弯曲乳杆菌、异型发酵的肠膜明串珠菌以及绿色魏斯菌等乳酸菌组成.在蒸煮阶段能够杀灭绝大多数微生物,腐败菌污染主要是在于后续的冷却、切片和真空包装阶段.采用超高压处理以及应用生物保护培养物,可以有效控制该类肉制品中的腐败微生物及部分致病微生物.     

超高压处理抑制低温烟熏火腿中的优势腐败菌

研究尝试应用微生物菌体总RNA提取代替DNA提取,进而通过反转录-PCR( RT-PCR),结合变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)技术,以期揭示超高压处理后低温烟熏火腿中腐败微生物的存活情况,探索RNA-DGGE手段判定超高压处理后微生物存活状态的可行性.分别以400 MPa和600 MPa的压力在室温(22℃)条件下,对包装后的烟熏火腿进行10 min超高压处理,未经高压处理样品作对照,于4℃冷藏条件下,贮藏1、15、30、60、90 d,直接提取样品中微生物的总RNA,对其进行RT-PCR和DGGE指纹图谱分析.DGGE指纹图谱显示,超高压处理对烟熏火腿中的优势腐败菌具有较强的抑制作用,且随压力的升高抑菌效应增强;超高压处理后烟熏火腿微生物种群结构变得单一,Weissella viridescens和Leuconostoc mesenteroides是超高压处理后烟熏火腿中的优势腐败菌.基于菌体总RNA提取的DGGE手段能够有效检测超高压处理后微生物的存活状况,揭示超高压对低温烟熏火腿中优势腐败微生物的抑菌效果.     

Detection of Leuconostoc strains at a meat processing plant using polymerase chain reaction

To simplify the labor-intensive conventional routine testing of samples to detect Leuconostoc at a meat processing plant, we developed polymerase chain reaction (PCR) primers specific for Leuconostoc from 16S rRNA gene sequences. These primers did not detect other common lactic acid bacteria such as Lactobacillus plantarum, Lact. sake, Lact. fermentum, Lact. acidophilus and Weissella viridescens. PCR with this primer detected all Leuconostoc species tested (Leu. mesenteroides subsp. mesenteroides, Leu. pseudomesenteroides, Leu. carnosum, Leu. lactic, Leu. citreum, Leu. amelibiosum, Leu. gelidum), except for Leu. fallax, and no other lactic acid bacteria on agarose gel electrophoresis. The method could identify areas contaminated with Leuconostoc in a large-scale industrial meat processing plant. Of 69 samples analyzed, 34 were positive for Leuconostoc according to the conventional culture method (isolation of LAB producing dextran) and PCR, whereas 29 were negative according to both. Six samples were culture-negative but positive by PCR. No false negative results were generated by PCR. The method is rapid and simple, is useful for routinely monitoring areas contaminated with Leuconostoc in meat processing plants, and could help to prevent the spoilage of meat products.     

超高压协同温度处理对绿色魏斯氏菌的失活动力学

为了建立超高压协同温度处理对绿色魏斯氏菌在低温烟熏火腿中的失活模型,采用40、45、50 ℃结合 200～500 MPa对接种108～109 CFU/mL绿色魏斯氏菌的烟熏火腿进行5、10、15、20、25、30 min的超高压处理,同 时做相同条件的离体实验进行对比。采用薄层平板计数,以时间为横坐标,亡菌数量级为纵坐标作失活曲线,选择 一级线性模型和非线性模型（Weibull和Logistic）进行模型的拟合,用模型的评价因子相关系数R2、准确因子Af和 均方差验证方程的拟合度。结果表明：经热压结合处理后,接种于烟熏火腿中的绿色魏斯氏菌比相同条件下离体状 态的绿色魏斯氏菌的失活更缓慢,可能因为火腿基质中存在一些大分子物质（如蛋白质、脂肪等）对菌体的保护作 用。Logistic方程最适合描述绿色魏斯氏菌在两种状态下的失活情况,为预测绿色魏斯氏菌在烟熏火腿中的失活提 供了理论依据。     

The spoilage microflora of cured, cooked turkey breasts prepared commercially with or without smoking

Lactobacillus sakei subsp. carnosus was predominant in the spoilage flora of sliced, vacuum-packed, smoked, oven-cooked turkey breast fillets which developed mild, sour spoilage flavors after 4 weeks storage at 4 degrees C. In contrast, Leuconostoc mesenteroides subsp. mesenteroides predominated in the spoilage flora of sliced, vacuum-packed, unsmoked, boiled turkey breast fillets from the same plant which were also stored at 4 degrees C. The spoilage flora of the unsmoked breasts grew faster than that of the smoked breasts and was more diverse. Lactobacillus sakei, Weissella viridescens and an atypical group of leuconostoc-like bacteria were also members of the unsmoked turkey breasts flora. Consequently, the unsmoked breasts spoiled after 2 weeks at 4 degrees C: the packs swelled and the meat developed strong sour odors and flavors and abundant slime. Except for the unidentified leuconostocs, which apparently survived boiling of the unsmoked turkey, all the spoilage organisms contaminated the meats during the slicing and vacuum packaging operations. From their biochemical reactions and cellular fatty acid profiles, the atypical leuconostocs were more closely related to Leuconostoc carnosum than W. viridescens. Carnobacteria and Brochothrix thermosphacta were present in relatively large numbers on the raw turkey, but were not numerous in the spoilage flora of the cooked, vacuum-packed meat products.     

Characterization and identification of lactic acid bacteria in "morcilla de Burgos"

A total of 176 lactic acid bacteria (LAB) isolated from a typical Spanish blood sausage called "morcilla de Burgos" were identified by means of phenotypic characteristics and 16S rDNA RFLP (ribotyping). LAB were isolated from "morcilla" of different producers and in different storage periods, which includes unpackaged, vacuum and modified atmosphere packaged "morcilla" and vacuum packed and pasteurised "morcilla". The knowledge of specific spoilage bacteria of "morcilla de Burgos" will be useful to design new preservation methods to extend the shelf-life of this product. Identification made according to phenotypic and biochemical characteristics shows the majority of the isolates were heterofermentative LAB (93.2%) and eight different bacterial groups could be distinguished (A-G). Weisella viridescens was the main species detected (42%). In addition, Leuconostoc spp. (23.9%), Weissella confusa (11.4%) and Lactobacillus fructosus (5.7%) species were found. Few strains were phenotypically misidentified as Lactobacillus sanfrancisco, Pediococcus spp., Lactobacillus sakei/curvatus and Carnobacterium spp. and 11 strains remained unknown. Most of the leuconostocs were identified as Leuconostoc mesenteroides and Leuconostoc carnosum species. Ribotyping shows a quite good correlation with phenotypic methods, although it has been possible to identify 15 different clusters. W. viridescens and leuconostocs were also the predominant LAB. Strains identified as W. confusa by phenotypic characteristics were resolved in W. confusa and Weissella cibaria by ribotyping. Neither Carnobacterium piscicola nor Lb. sanfrancisco were identified by means of genotypic method. All Lb. fructosus strains and some more included in different phenotypic groups (17 strains in total) could not be associated with any reference strain (cluster VII).     

Leuconostoc carnosum associated with spoilage of refrigerated whole cooked hams in Greece

A polyphasic taxonomic approach was used to identify a major atypical group of gas-forming, arginine-negative lactic acid bacteria associated with spoilage of whole (nonsliced) refrigerated (4 degrees C) cooked hams produced in two Greek industrial meat plants. Biochemical characterization revealed that the ham isolates shared their phenotypic properties with Leuconostoc carnosum, Weissella viridescens, and Weissella hellenica. However, gas chromatographic analysis of cellular fatty acids clearly differentiated the ham isolates from the Weissella spp. None of the isolates contained eicosenoic acid (n-C20:1), which is typically synthesized by W. viridescens, but all strains contained high amounts of C 19cycl acid, which is absent in W. hellenica and has been found in trace amounts in W. viridescens. All strains had similar cellular fatty acid profiles, which were qualitatively similar to those of the cellular fatty acids of L. carnosum. In addition to the phenotypic and chemotaxonomic tests, three representative isolates were studied using a lactic acid bacteria database, which employs 16S and 23S HindIII restriction fragment length polymorphism patterns as operational taxonomic units in a numerical analysis. The isolate patterns were identical to those of the L. carnosum type strain, NCFB 2776T. Based on the polyphasic taxonomic approach, the dominating lactic acid bacteria group was identified as L. carnosum.     

Characterisation and biochemical properties of predominant lactic acid bacteria from fermenting cassava for selection as starter cultures

A total of 375 lactic acid bacteria were isolated from fermenting cassava in South Africa, Benin, Kenya and Germany, and were characterised by phenotypic and genotypic tests. These could be divided into five main groups comprising strains of facultatively heterofermentative rods, obligately heterofermentative rods, heterofermentative cocci, homofermentative cocci and obligately homofermentative rods, in decreasing order of predominance. Most of the facultatively heterofermentative rods were identified by phenotypic tests as presumptive Lactobacillus plantarum-group strains, which also comprised the most predominant bacteria (54.4% of strains) isolated in the study. The next predominant group of lactic acid bacteria (14.1% of total isolates) consisted of obligately heterofermentative rods belonging either to the genus Lactobacillus or Weissella, followed by the heterofermentative cocci (13.9% of isolates) belonging to the genera Weissella or Leuconostoc. Homofermentative cocci were also isolated (13.3% of isolates). Biochemical properties such as production of alpha-amylase, beta-glucosidase, tannase, antimicrobials (presumptive bacteriocin and H(2)O(2)-production), acidification and fermentation of the indigestible sugars raffinose and stachyose, were evaluated in vitro for selection of potential starter strains. A total of 32 strains with one or more desirable biochemical properties were pre-selected and identified using rep-PCR fingerprinting in combination with 16S rRNA sequencing of representative rep-PCR cluster isolates. Of these strains, 18 were identified as L. plantarum, four as Lactobacillus pentosus, two each as Leuconostoc fallax, Weissella paramesenteroides and Lactobacillus fermentum, one each as Leuconostoc mesenteroides subsp. mesenteroides and Weissella cibaria, while two remained unidentified but could be assigned to the L. plantarum-group. These strains were further investigated for clonal relationships, using RAPD-PCR with three primers, and of the 32 a total of 16 strains were finally selected for the development as starter cultures for Gari production.     

水牛乳中可培养乳酸菌多样性分析

采用传统分离培养方法,从三品杂交生水牛奶混合样品中,分离出105株乳酸菌,通过形态、生理生化、API细菌鉴定系统及16S rDNA基因序列分析方法对各菌株属种进行鉴定。16S rRNA序列分析结果显示,105株菌共分为5个属8个种,呈现较为丰富的乳酸菌多样性,具体数量分布为乳酸乳球菌21株,植物乳杆菌19株,格氏乳球菌17株,乳明串珠菌13株,食窦魏斯氏菌11株,肠膜明串珠菌8株,类肠膜魏斯氏菌6株,嗜热链球菌5株,糊精乳杆菌5株。由此可知,水牛乳中可培养乳酸菌优势菌群的主次关系为：乳酸乳球菌（Lactococcus lactis）＞植物乳杆菌（Lactobacillus plantaru）＞格氏乳球菌（Lactococcus garvieae）＞乳明串珠菌（Leuconostoc lactis）＞食窦魏斯氏菌（Weissella cibaria）,此为后续开发水牛乳中优势乳酸菌资源提供了良好的理论基础。