广州市售酸奶中乳酸菌的耐药性评估

研究广州市售酸奶中乳酸菌对不同种类抗生素的药物敏感性。从市售酸奶中分离并鉴定乳酸菌,采用纸片扩散法检测分离菌株对18种抗生素的药物敏感性;E-test方法对耐药菌株进行最低抑菌浓度测定。从31份市售保质期内商标注册酸奶中分离鉴定出48株乳酸菌,其中25株嗜热链球菌(Streptococcus thermophilus)、20株保加利亚乳杆菌(Lactobacillus bulgaricus)、3株副干酪乳杆菌(Lactobacillus paracasei)。48株乳酸菌对卡那霉素、链霉素均耐药;对头孢他啶、头孢噻肟、红霉素均敏感;对环丙沙星、庆大霉素、万古霉素等呈现出不同程度的耐受性;79.2%(38/48)表现出多重耐药性;E-test分析表明,除了保加利亚乳杆菌MR10的MIC为256μg/mL以外,所有万古霉素耐药菌株对万古霉素的MIC均大于1024μg/mL。不同品牌酸奶分离菌株对抗生素具有不同的耐受性,均有多重耐受性出现,应加强其耐药性连续监测及安全评价。     

高加索酸奶中乳酸菌的分离与鉴定

从自然发酵的5份酸奶样品中,通过平板划线等方法分离筛选乳酸菌。经形态特征,生理生化特性及糖发酵试验等,筛选到12株乳酸菌,分别为:乳杆菌7株,其中:3株德氏乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp.bulgaricus),3株瑞士乳杆菌(Lactobacillus hel-veticus),1株罗伊氏乳杆菌(Lactobacillus reuteri);乳酸球菌5株,包括3株嗜热链球菌(Streptococcus thermophilus),2株乳酸乳球菌乳脂亚种(Lactococcus lactis subsp.Cremoris)。     

基于特征风味定向筛选酸奶用乳酸菌

针对国内酸奶发酵剂成本高,菌剂被国外垄断的问题。本实验基于特征风味物质乙醛和双乙酰,从市售酸奶产品和菌剂中定向筛选出高风味物质含量菌株,经复配和感官评定,得到适合生产菌株。实验共分离鉴定出58株乳酸菌,包括嗜酸乳杆菌(9株)、鼠李糖乳杆菌(2株)、德氏乳杆菌(4株)、嗜热链球菌(39株)、乳酸乳球菌乳酸亚种/乳酸乳球菌(4株)。产乙醛能力呈现出德氏乳杆菌嗜热链球菌乳酸乳球菌嗜酸乳杆菌鼠李糖乳杆菌,最高达到25.57μg/g。产双乙酰能力呈现出鼠李糖乳杆菌嗜酸乳杆菌嗜热链球菌乳酸乳球菌德氏乳杆菌,最高达到3.57μg/g。德氏乳杆菌和鼠李糖乳杆菌产酸能力较强。菌株复配后大多风味物质含量提高,在一定条件下,呈现出乙醛/双乙酰比例越高,乙醛含量越高,感官风味越好。     

酸奶中乳酸菌的药物敏感性分析

目的:分析和鉴定不同产地酸奶中乳酸菌对一些常用抗生素的敏感性,指导合理使用抗菌药物与合理使用乳酸菌发酵制品。方法:利用改良的MRS培养基从各地酸奶中分离获得乳酸菌,然后采用琼脂稀释法,对分离获得的18株保加利亚乳杆菌和28株嗜热链球菌对11种常用抗生素的药物敏感性进行检测和分析。结果:同一菌株对不同的抗生素具有不同的药物敏感性,不同菌株对同一种抗生素也显示出不同的敏感性。结论:乳酸菌对常用抗生素的多重耐药已成为普遍现象,不同来源的菌株表现出了不同的耐药性。     

利用16S rDNA序列及tuf-RFLP鉴定蒙古国发酵乳中的乳酸菌

运用16S rDNA序列分析和tuf-RFLP技术对采于蒙古国扎布汗省的25份发酵乳样中分离出的110株乳酸菌进行鉴定。首先将分离的110株乳酸菌的16S rRNA基因进行扩增,测序并构建系统发育树,初步鉴定为41株嗜热链球菌,40株瑞士乳杆菌,11株德氏乳杆菌保加利亚亚种,2株发酵乳杆菌,1株乳明串珠菌,2株肠膜明串肠膜亚种,1株乳酸乳球乳酸亚种和12株属于干酪族的菌株。由于干酪乳杆菌族的16S rDNA序列差异很小,故采用tuf-RFLP技术对这12株进行了进一步的验证,通过分离菌株与模式菌株tuf-RFLP图谱的比较分析,结果表明这12株菌均为干酪乳杆菌。     

泡菜发酵乳酸菌的分离鉴定及耐药性分析

对吉林地区朝鲜族自制泡菜中的发酵乳酸菌进行分离鉴定,并对菌株耐药情况进行分析,以期为泡菜发酵菌的风险评估奠定基础。采集吉林省多个不同地区的泡菜制品,通过MRS培养基分离,16S rDNA测序,分析其乳酸菌种类及数量,同时采用平板琼脂稀释法进一步检测了分离菌对10 种临床常用抗生素的药物敏感性。结果表明,从15 份不同厂家泡菜中共分离得到34 株乳酸菌,分别为植物乳杆菌12 株,短乳杆菌5 株,鼠李糖乳杆菌3 株,干酪乳杆菌2 株,清酒杆菌2 株,肠膜明串珠菌2 株,嗜热链球菌2 株,食窦魏斯氏乳杆菌1 株,弯曲乳杆菌1 株,棒状乳杆菌1 株,嗜酸乳杆菌1 株,坚强肠球菌1 株,Lactobacillus namurensis 1 株。药敏实验结果表明,所有菌株对氨苄西林和氯霉素敏感,对其他抗生素均有不同程度的耐药性,其中对环丙沙星耐药率最高。     

传统发酵面团的菌相分析

对传统发酵面团的主要微生物菌群进行了分析,分离得到12株酵母菌,3株乳酸菌,16株醋酸菌,并对分离菌种进行了形态观察,生理生化反应实验,对菌种进行了初步鉴定。所得酵母菌主要为酵母属酵母,汉逊酵母属及毕赤氏酵母;乳酸菌主要为有嗜热链球菌,德氏乳杆菌保加利亚亚种及一种未知乳杆菌;醋酸菌主要为醋化醋杆菌,巴氏醋杆菌,汉氏醋杆菌及液化醋杆菌。     

西藏传统发酵奶制品中乳酸菌多样性分析及产胞外多糖菌株筛选

从22份西藏地区传统发酵奶制品中分离到113株乳酸菌,16S rDNA鉴定结果显示它们分属于6个属,包括乳杆菌属、链球菌属、明串珠菌属、肠球菌属、乳球菌属和芽孢杆菌属。其中乳杆菌属有12个种,德氏乳杆菌保加利亚亚种和发酵乳杆菌为优势菌种,另外还包括到1株格氏乳杆菌和1株卷曲乳杆菌。采用菌落拉丝并结合检测胞外多糖产量的方法,筛选到16株乳酸菌可在脱脂乳中产生胞外多糖。挑取其中产量较高的3株进行发酵性能测试,发现菌株德氏乳杆菌保加利亚亚种1 576的产酸速率适中、后酸化稳定,并能形成质构稠厚的发酵乳。     

水牛乳中乳酸菌的筛选

从自然发酵的水牛乳中分离、筛选乳酸菌株,用16SrRNA寡核苷酸序列分析方法对分离得到的菌株进行鉴定,结果得到5株乳酸菌株,分别为德氏乳杆菌保加利亚亚种3株、嗜热链球菌1株、乳酸乳球菌乳酸亚种1株,通过组合发酵试验,得到4组优良的混合发酵菌种。     

开菲尔粒中乳酸菌的分离与鉴定

从4个不同来源的开菲尔粒中分离得到40株乳酸菌，并对这40株乳酸菌进行了鉴定．结果为德氏乳杆菌8株，德氏乳杆菌保加力亚种6株，高加索奶乳杆菌2株，嗜酸乳杆菌1株，粪肠球菌7株，屎肠球菌2株，嗜热链球菌1株，乳脂链球菌3株。     

Interactions among lactic acid starter and probiotic bacteria used for fermented dairy products

Interactions among lactic acid starter and probiotic bacteria were investigated to establish adequate combinations of strains to manufacture probiotic dairy products. For this aim, a total of 48 strains of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium spp. (eight of each) were used. The detection of bacterial interactions was carried out using the well-diffusion agar assay, and the interactions found were further characterized by growth kinetics. A variety of interactions was demonstrated. Lb. delbrueckii subsp. bulgaricus was found to be able to inhibit S. thermophilus strains. Among probiotic cultures, Lb. acidophilus was the sole species that was inhibited by the others (Lb. casei and Bifidobacterium). In general, probiotic bacteria proved to be more inhibitory towards lactic acid bacteria than vice versa since the latter did not exert any effect on the growth of the former, with some exceptions. The study of interactions by growth kinetics allowed the setting of four different kinds of behaviors between species of lactic acid starter and probiotic bacteria (stimulation, delay, complete inhibition of growth, and no effects among them). The possible interactions among the strains selected to manufacture a probiotic fermented dairy product should be taken into account when choosing the best combination/s to optimize their performance in the process and their survival in the products during cold storage.     

市售酸奶中乳酸菌耐药性及耐药基因的检测

为探讨酸奶中乳酸菌所携带耐药基因对人类健康的潜在影响,对市售酸奶中的乳酸菌进行分离和鉴定,并通过药物敏感性实验确定菌株的耐药谱,同时利用聚合酶链式反应（polymerase chain reaction,PCR）扩增技术调查链霉素、庆大霉素、磺胺类和四环素等耐药基因的分布情况。结果表明：25 份市售酸奶样品中共分离得到56 株乳酸菌,包括26 株德氏乳杆菌保加利亚亚种、3 株植物乳杆菌、2 株嗜酸乳杆菌以及25 株嗜热链球菌。药敏结果显示,31 株乳杆菌对链霉素（87.1%）、庆大霉素（80.6%）、环丙沙星（74.2%）和四环素（61.3%）的耐药率较高,对头孢菌素类则较为敏感;而25 株嗜热链球菌同样对链霉素的耐药率最高,达76.0%;其次分别为万古霉素（32.0%）、环丙沙星（32.0%）和四环素（20.0%）。56 株乳酸菌中共检出5 种不同的耐药基因,分别为链霉素耐药基因ant(6)（检出率1.8%）、庆大霉素耐药基因aac(6')-aph(2')（检出率7.1%）、四环素耐药基因tetM（检出率5.4%）以及磺胺类耐药基因sulⅠ（检出率14.3%）和sulⅡ（检出率1.8%）。受试的乳酸菌中共有13 株检出耐药基因,其中有4 株携带两种不同的耐药基因。长期以来被认为安全并广泛应用于发酵食品领域的乳酸菌可能成为潜在的耐药基因贮存库。     

Production of pyroglutamic acid by thermophilic lactic acid bacteria in hard-cooked mini-cheeses

Pyroglutamic acid is present in high amounts (0.5g/ 100g) in many cheese varieties-and particularly in extensively ripened Italian cheeses such as Grana Padano and Parmigiano Reggiano. An in vivo model system for cooked mini-cheese production and ripening acceleration was set up to demonstrate the ability of thermophilic lactic acid bacteria, used as a starter, to produce pyroglutamic acid (pGlu). In mini-cheeses stored at 38 and 30 degrees C for up to 45 d, all starters tested produced different amounts of pGlu. In descending order of pGlu production, the bacteria analyzed were: Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactobacillus delbrueckii subsp. lactis. Evidence for the presence of glutamine to pGlu cyclase activity in lactic acid bacteria was provided. Cell lysates obtained from cultures of L. helveticus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, and S. thermophilus showed the ability to cyclize glutamine to pGlu, resulting in processing yields from 1.4 to 30.3%, depending on the subspecies. Formation of pGlu from free glutamine appeared to be similar to that observed using a glutamine-glutamine dipeptide substrate. Under the experimental conditions applied, pGlu aminopeptidase activity was only detected in L. helveticus. Thus, pGlu formation in long-ripened cooked cheese may depend on the activity of thermophilic lactic acid bacteria.     

内蒙古酸马奶中乳酸菌多样性的研究

采用16S rRNA基因全序列测定和聚类分析技术,对酸马奶中的乳酸菌进行了准确鉴定并构建了乳酸菌的系统发育树.然后对乳酸菌菌群进行了多样性分析,结果显示,酸马奶中的优势乳酸菌分别为:Lactobacillus plantarum(10%),Lactobacillus brevis(8%),Lactobacillus casei(7.8%),Enterococcus faecium(17%),Enterococcus faecalis(14%),Lactococcuslactis(19%),Lactobacillus acidlophilus(5%),Lactobacillus paracasei(2%),Lactobacillus delbrueckii subsp.bulgaricus(4%),Lactobacillus helveticus(4%),Enterococcus durans(4%),Leuconostoc mesenteroides (4%),Leuconostoc garlicum(1%),Streptococcus thermophilus(1%).     

DNA fingerprinting of thermophilic lactic acid bacteria using repetitive sequence-based polymerase chain reaction

DNA fingerprints of lactic acid bacteria were generated by polymerase chain reaction using a primer based on the repetitive elements found in the genome of Streptococcus pneumoniae (BOX-PCR). The method made it possible to identify 37 isolates from raw milk. industrial starters and yogurt. Differentiation at species, subspecies and strain level was possible for Lactobacillus delbrueckii subsp. lactis, Lb. delbrueckii subsp bulgaricus and Str. thermophilus. BOX-PCR was also applied to studying the strain composition of a starter culture and the direct detection of strains in commercial fermented milk.     

不同乳酸菌对椰子水饮料的发酵特性研究

以椰子水为原料,利用6株乳酸菌:发酵乳杆菌L20、面包乳杆菌32-2-2、短乳杆菌64-1、植物乳杆菌A33、嗜热链球菌IFFI6038和德氏乳杆菌保加利亚亚种CICC6045发酵椰子水制备饮料,分析测定发酵椰子水主要成分、抑菌活性和感官评价的变化。试验结果表明,发酵48 h时,面包乳杆菌32-2-2发酵椰子水饮料总酸含量最高,达10.85 g/L;植物乳杆菌A33发酵椰子水饮料pH值最低为3.1;德氏乳杆菌保加利亚亚种CICC6045发酵椰子水饮料还原糖含量最低为0.03 g/L;植物乳杆菌A33的抑菌谱最广;抑菌活性与pH值和乳酸含量呈显著相关性。植物乳杆菌、嗜热链球菌和德氏乳杆菌保加利亚亚种发酵椰子水感官评分较高。     

Proteolytic profiles and angiotensin-I converting enzyme and alpha-glucosidase inhibitory activities of selected lactic acid bacteria

ABSTRACT:  This study was conducted to examine the growth, proteolytic profiles as well as angiotensin-I converting enzyme (ACE) and α-glucosidase (α-glu) inhibitory potentials of selected strains of lactic acid bacteria (LAB). Two strains each of yogurt bacteria ( Streptococcus thermophilus —1275 and 285, and Lactobacillus delbrueckii ssp. bulgaricus —1092 and 1368), and probiotics ( L. acidophilus —4461 and 33200, and L. casei —2607 and 15286, and 1 strain of Bifidobacterium longum 5022), were cultivated in reconstituted skim milk (RSM) at 37 °C and their proteolytic profiles and ACE as well as α-glu inhibitory activities were determined. Among all the strains of lactic acid bacteria studied, yogurt bacteria grew very well, with the exception of L. delbrueckii ssp. bulgaricus 1368 which showed a slower growth during the initial 3 h of incubation. The growth pattern corresponded well with the decrease in pH for the organisms. All the organisms showed an increase in proteolysis with time. The variations in proteolytic capabilities translated into corresponding variations in ACE inhibitory potential of these organisms. Bifidobacterium longum 5022 showed the highest ACE inhibitory potential followed by L. delbrueckii ssp. bulgaricus 1368, L. casei 15286, S. thermophilus 1275, and L. acidophilus 4461. Organisms with high intracellular enzymatic activities grew well. Also, aminopeptidases of strains of L. acidophilus 4461 and S. thermophilus 1275 that could better utilize proline containing substrates showed enhanced ACE inhibitory potential. Lactic acid bacteria possessed the ability to inhibit α-glu activity, which endowed them an antidiabetic property as well.     

酸奶菌株分批发酵放大研究-从2.5L 到15L 发酵罐

2.5L发酵罐实验数据基础上,利用经验放大法将德氏乳杆菌保加利亚亚种KLDS 1.9201和唾液链球菌嗜热亚种KLDS 3.021 分批发酵结果从小试放大中试。结果表明,如果确定若干优化发酵参数,包括搅拌转速50～100r/min和溶氧浓度不高于5% 等,可将酸奶菌株发酵结果较成功地从2.5L 罐放大到15L 罐规模。KLDS 1.9201 在15L 罐中对数生长期间的初糖转化率达到了80.3%,乳酸浓度为49g/L,确定其发酵终点为12h,此时活菌数为2.2 × 109CFU/ml;KLDS 3.0201 在15L 罐中的初糖转化率达到了80.2%,乳酸浓度为39g/L,确定其最佳收获期为发酵8h,活菌数达到4.9 × 109CFU/ml。     

Characterisation of lactic acid bacteria isolated from fermented milk "laban"

The technological properties of 96 lactic acid bacteria isolated from Lebanese traditional fermented milk "laban" were characterised. They were classified by phenotypic and biochemical analyses as Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, thus indicating that laban is a fermented milk similar to yogurt. Most strains of L. bulgaricus (87.5%) exhibited a high acidification activity, whereas strains of streptococci showed low acidification ability. 33.3% of streptococci strains and 25% of lactobacilli strains displayed similar acidification performances as European strains. Results obtained for syneresis, texture and rheological parameters led us to consider that isolated strains were not low polymer-producing strains. Some of them displayed interesting characteristics such as low syneresis and high values for rheological parameters. The major flavour compounds found in pure cultures were acetaldehyde, acetone, 2-butanone, dimethyl disulfide, acetoin, 2,3-butanedione, 2,3-pentanedione, and acetic, hexanoic and butanoic acids. Acetaldehyde (7.4%) and organic acids (48.3%) were mainly produced by L. bulgaricus strains, whereas streptococci cultures contained high relative levels of 2,3-butanedione and acetoin, which represented around 82% of the total flavour compounds. Finally, strains isolated from laban samples exhibited different technological properties than those used in yogurt production, thus conferring specific characteristics to this product.