自然发酵牛乳中乳酸菌和酵母菌的活菌动态变化

构建三批次各100 kg牛乳自然发酵体系,通过分析乳酸菌和酵母菌的活菌动态变化,对两者间的相互作用进行研究。结果表明,在（20±1）℃的自然发酵温度下,持续发酵56 h,pH值下降至4.6～5.0,发酵牛乳逐渐分层并形成上层奶嚼口和下层凝乳。连续三个批次发酵牛乳中奶嚼口和凝乳的乳酸菌平均活菌数分别为（12.02±1.21）lg（CFU/mL）和（11.54±1.30）lg（CFU/mL）,差异不显著（P＞0.05）;而凝乳中酵母菌平均活菌数[（6.39±0.60）lg（CFU/mL）]显著低于奶嚼口[（4.56±0.30）lg（CFU/mL）]（P＜0.05）。活菌动态变化表明,在牛乳自然发酵过程中,乳酸菌的生长具有延迟期、对数期、稳定期三个典型阶段,而酵母菌没有明显的对数期,随着乳酸菌的生长反而抑制酵母菌的生长。     

Enumeration and identification of yeasts isolated from Zimbabwean traditional fermented milk

The yeasts in 30 samples of the Zimbabwean traditional fermented milk, amasi, taken from farms, households and milk collection centres were enumerated and identified. The yeast counts ranged from <2 to 8.08 log cfu g⁻¹. Yeast isolates were identified using the API ID 32°C test kits and the simplified identification method (SIM) as well as with reference to the standard taxonomic keys. From the 30 samples, a total of 20 different yeast species were identified. Saccharomyces (S) cerevisiae (22 isolates), Candida (C) lusitaniae (11), C. colliculosa (7) and S. dairenensis (7) were the predominant species identified. Dekera (Dek.) bruxillensis, C. lipolytica and C. tropicalis were identified less often. Seven of the S. cerevisiae isolates were able to assimilatedl-lactate. The strain of C. kefyr isolated could assimilate lactose anddl-lactate, but not citrate. The analysed amasi samples contained a wide variety of yeasts, but only a few species predominated and these could possibly contribute to the characteristics of the fermented milk in the 48 h fermentation.     

传统发酵酸牛奶中乳酸菌多态性分析

以从内蒙古呼伦贝尔市牧区采集的1份传统发酵酸牛奶样品为研究对象,对其进行乳酸菌的分离鉴定。通过传统纯培养法分离出17株菌,并对17株菌进行16SrDNA序列分析、多位点pheS序列分析和生理生化鉴定,鉴定的结果为11株乳酸乳球菌乳酸亚种、1株格式乳球菌、1株粪肠球菌、2株植物乳杆菌植物亚种及2株弯曲乳杆菌。乳酸乳球菌乳酸亚种为优势菌(占总分离菌株的64.7%)。     

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.     

西藏牦牛发酵乳中乳酸菌及酵母菌的特性与相互作用

研究了西藏牦牛发酵乳中的优势菌株:发酵乳杆菌166A、乳酒假丝酵母37、酿酒酵母100、郎比克假丝酵母58在牛乳中的生长特性。探讨了发酵乳杆菌与酵母菌间的相互作用,在发酵过程中3株酵母菌均对发酵乳杆菌166A的生长有促进作用,发酵乳杆菌166A能抑制乳酒假丝酵母37的生长。酵母菌与发酵乳杆菌166A共同接种有利于保持产品冷藏期间活菌数的稳定,菌株之间可能存在共生作用。     

Biodiversity of lactic acid bacteria isolated from fermented milk products in Xinjiang,China

Fermented dairy products have been produced and consumed for thousands of years in Xinjiang, China. In this study, traditional culture-dependent methods and 16S rRNA gene analyses were performed to analyze the composition of lactic acid bacteria (LAB) from 86 samples of traditional fermented dairy products collected from four regions of Xinjiang. Quantitative polymerase chain reaction (qPCR) was used to quantify Bifidobacterium and Lactobacillus species. The LAB isolates (N = 705) were identified as belonging to seven genera and 26 species or subspecies. The predominant species of all isolates were Lactobacillus (Lb.) delbrueckii subsp. bulgaricus, Lb. fermentum, and Lb. helveticus. The LAB counts of these samples ranged from 5.35 to 10.06 Log CFU/mL. The bacterial counts of seven lactobacilli species and Bifidobacterium ranged from 0.98 ± 0.14 Log CFU/mL (for Lb. sakei from fermented mare milk) to 9.91 ± 0.17 Log CFU/mL (for Lb. helveticus from fermented mare milk). While the microbiota was significantly different between yak and cow milk, there was no significant difference between the Tex and Zhaosu counties. In conclusion, LAB communities in traditional fermented dairy products are complex and differ among local regions within Xinjiang.     

青海省海西地区传统发酵山羊乳中乳酸菌的分离及鉴定

从采自青海省海西地区牧民家庭的11份以山羊奶为主要原料的传统发酵乳中分离得到33株乳酸菌，经过形态特征．生理生化特性，糖发酵试验和乳酸旋光性的测定，将这33株乳酸茵初步鉴定为7株Lldelbrueckiisubsp．Lactis，4株L．delbrueckii subsp．bulgaricus.3株L．acidophilus，1株L，coryniformis subsp，corynformis，12株S．thermophilus，Ped．日cidilaaici 3株，Ped．Unnaeequi 2株，Leu．mesenteriodes subsp.mesentemiodes 1株，其中，杆菌占所分离菌株的45．45％，球菌占所分离菌株的54．55％。     

发酵乳中酵母菌和乳酸菌生长的相互影响

探讨了乳酸菌和酵母菌之间的相互作用。在发酵过程中,酿酒酵母对乳酸菌的生长有抑制作用。乳酸菌能促进酿酒酵母和马克思克鲁维酵母的生长。酿酒酵母和乳酸菌共同接种有利于保持产品冷藏期间活菌数的稳定,菌株之间可能存在共生作用。      

The role of interaction between yeasts and lactic acid bacteria in African fermented milks: a review

Yeasts are present in indigenous African fermented milks in numbers up to log 8 cfu g(-1), together with a varied lactic acid bacteria (LAB) flora, and therefore potentially contribute to product characteristics. However, interaction between yeasts and LAB in these products has received little notice. In studies of indigenous fermented milk in Zimbabwe and Uganda, many samples contained more than one species of yeast, but Saccharomyces cerevisiae was most commonly isolated. Other frequent isolates were other species of Saccharomyces and several species of Candida. Most yeast isolates were lactose-negative but usually galactose-positive. Some strains assimilated lactate and citrate. The growth in milk of strains of yeasts and LAB, isolated from naturally soured milk in Zimbabwe, and their interaction when selected pairs of strains were grown together has been studied. Interactions were shown by the significantly different amounts of certain metabolites produced, such as acetaldehyde and malty aldehydes, when co-cultures were compared to pure cultures. Preliminary sensory acceptance tests did not show, however, that milks made from a co-culture with Candida kefyr and LAB were preferable to the pure LAB culture. Further work is still needed to elucidate the reactions that may be taking place in fermented milk between varying LAB and yeast populations. The potential for use as starter cultures depends on various aspects, including the final product being prepared. The role of other microorganisms in naturally fermented milk also needs to be studied.     

自然发酵三樱椒中乳酸菌的分离及鉴定

为获得适用于三樱椒发酵的乳酸菌,该研究采用钙溶圈法从自然发酵三樱椒中分离乳酸菌,通过形态观察和分子生物学技术对其进行菌种鉴定,并对其生长、产酸、耐酸特性进行分析。结果表明,从自然发酵三樱椒中共分离纯化得到19株乳酸菌,经鉴定,16株为植物乳杆菌（Lactobacillus plantarum）,3株为赫轮魏斯氏菌（Weissella hellenica）。培养初期,赫轮魏斯氏菌生长比植物乳杆菌快,但培养12 h后植物乳杆菌比赫轮魏斯氏菌生长速度更快;植物乳杆菌产酸能力优于赫轮魏斯氏菌,培养21 h后,植物乳杆菌发酵液的pH值稳定在3.7,赫轮魏斯氏菌发酵液pH值稳定在4.3,其中植物乳杆菌PLP-1产酸能力最好;植物乳杆菌的耐酸性优于赫轮魏斯氏菌,在pH值＜4.0的培养条件下赫伦魏斯氏菌生长受到严重抑制,而植物乳杆菌可以在pH值为3.0的培养基中生长,其中植物乳杆菌PLP-1耐酸性最好。     

从风干肠中分离出的三株乳酸菌发酵性能的研究

对从自然发酵风干肠中分离出的三株乳酸菌的发酵性能进行研究,主要包括菌种的耐盐性试验、耐亚硝酸盐试验、生长温度的测定、产酸速率的测定、蛋白质及脂肪降解活性试验、拮抗性试验。结果表明:分离乳酸菌1号菌的耐热性强,可能在烘烤后继续发挥产酸效力,因此不适合作为发酵剂使用;2号菌、3号菌则可在6%的食盐浓度下和0.15g/kg的亚硝酸钠下生长,在65℃以上完全不生长,产酸速度快,不分解蛋白质和脂肪,相互之间无拮抗作用,完全符合肉制品发酵剂的要求。     

核桃乳酸菌饮料的研制

主要通过实验筛选出在核桃、大豆等植物蛋白中适宜生长且产酸产香良好的乳酸菌种,研究出复合植物蛋白原料的最佳配比与最适接种量,经过发酵、调配,研制出品质达标的核桃乳酸菌饮料。     

Taxonomic and molecular characterization of lactic acid bacteria and yeasts in nunu,a Ghanaian fermented milk product

Produced from raw unpasteurized milk, nunu is a spontaneously fermented yoghurt-like product made in Ghana and other parts of West Africa. Despite the importance of nunu in the diet of many Africans, there is currently only limited information available on the microorganisms associated with nunu processing. With the aim of obtaining a deeper understanding of the process and as a first step towards developing starter cultures with desired technological properties for nunu production, a microbiological characterization of nunu processing in three different towns in the Upper East region of Ghana, namely Bolgatanga, Paga and Navrongo, was carried out. Lactic acid bacteria (LAB) and yeasts associated with nunu processing were isolated and identified using a combination of pheno- and genotypic methods including morphological and carbohydrate fermentation tests, (GTG)₅-based rep-PCR, multiplex PCR, and 16S and 26S rRNA gene sequencing. The LAB counts during nunu processing increased from 4.5 ± 0.4 log cfu/ml at 0 h to 8.7 ± 1.8 log cfu/ml at 24 h of fermentation while yeasts counts increased from 2.8 ± 1.2 log cfu/ml at 0 h to 5.8 ± 0.5 log cfu/ml by the end of fermentation. Lactobacillus fermentum was the dominant LAB throughout the fermentations with Lactobacillus plantarum and Leuconostoc mesenteroides playing prominent roles during the first 6–8 h of fermentation as well. Less frequently isolated LAB included Lactobacillus helveticus, Enterococcus faecium, Enterococcus italicus, Weissella confusa and a putatively novel Lactococcus spp. The yeasts involved were identified as Candida parapsilosis, Candida rugosa, Candida tropicalis, Galactomyces geotrichum, Pichia kudriavzevii and Saccharomyces cerevisiae with P. kudriavzevii and S. cerevisiae being the dominant yeast species.     

Enhancement of survival of probiotic and non-probiotic lactic acid bacteria by yeasts in fermented milk under non-refrigerated conditions

The effects of yeasts on the survival of probiotic and non-probiotic lactic acid bacteria (LAB) were studied in fermented milk under non-refrigerated conditions (30 °C) with a view to develop ambient-stable fermented milk with live LAB. Five yeasts tested (Saccharomyces bayanus, Williopsis saturnus var. saturnus, Yarrowia lipolytica, Candida kefyr and Kluyveromyces marxianus) enhanced the survival of Lactobacillus bulgaricus (but not Streptococcus thermophilus) in a mixed yoghurt culture in yoghurt by ~ 10² to 10⁵-fold. Seven yeasts examined (Candida krusei, Geotrichum candidum, Pichia subpelliculosa, Kloeckera apiculata, Pichia membranifaciens, Schizosaccharomyces pombe and Y. lipolytica) improved the survival of Lactobacillus rhamnosus in fermented milk by ~ 10³ to 10⁶-fold. W. saturnus var. saturnus enhanced the survival of Lactobacillus acidophilus, L. rhamnosus (probiotic) and Lactobacillus reuteri by up to 10⁶-fold, but the same yeast failed to improve the survival of Lactobacillus johnsonii (probiotic), S. thermophilus and L. bulgaricus in fermented milk. These results provide definitive evidence that yeasts possess stability-enhancing effects on LAB and that the specific effects of yeasts on LAB stability vary with yeasts as well as with LAB. However, the molecular mechanism of such interaction of yeasts with LAB remains to be found.     

Growth characteristics of Candida kefyr and two strains of Lactococcus lactis subsp. lactis isolated from Zimbabwean naturally fermented milk.

Lactic acid bacteria (LAB) and yeasts constitute part of the microflora in Zimbabwean traditional fermented cows' milk, amasi. The present study was carried out to investigate the growth characteristics of Candida kefyr 23, Lactococcus lactis subsp. lactis biovar. diacetylactis C1 and L. lactis subsp. lactis Lc261, previously isolated from amasi, in ultrahigh temperature (UHT)-treated cows' milk. The strains were inoculated into the UHT milk as both single and yeast     

新疆传统发酵乳品中乳酸菌与酵母菌生长的相互影响

对新疆传统发酵乳酪乳清中的优势菌株:马乳酒样乳杆菌、乳酸乳球菌、瑞士乳杆菌、植物乳杆菌、东方伊莎酵母菌在驼乳中的生长特性进行了研究。探讨了4株乳酸菌与东方伊莎酵母菌之间的相互作用。结果表明,在发酵过程中东方伊莎酵母菌显著促进马乳酒样乳杆菌和乳酸乳球菌的生长(p0.05),对瑞士乳杆菌的生长促进作用不明显(p0.05)。同时,4株乳酸菌抑制东方伊莎酵母菌的生长(p0.05)。此外,乳酸菌与东方伊莎酵母菌共同接种发酵有利于保持发酵乳冷藏期间活菌数的稳定及缓解乳酸菌的过度产酸。综上所述:乳酸菌与酵母菌之间可能存在相互作用的关系。     

新疆传统发酵乳品中乳酸菌与酵母菌生长的相互影响

对新疆传统发酵乳酪乳清中的优势菌株:马乳酒样乳杆菌、乳酸乳球菌、瑞士乳杆菌、植物乳杆菌、东方伊莎酵母菌在驼乳中的生长特性进行了研究。探讨了4株乳酸菌与东方伊莎酵母菌之间的相互作用。结果表明,在发酵过程中东方伊莎酵母菌显著促进马乳酒样乳杆菌和乳酸乳球菌的生长(p<0.05),对瑞士乳杆菌的生长促进作用不明显(p>0.05)。同时,4株乳酸菌抑制东方伊莎酵母菌的生长(p<0.05)。此外,乳酸菌与东方伊莎酵母菌共同接种发酵有利于保持发酵乳冷藏期间活菌数的稳定及缓解乳酸菌的过度产酸。综上所述:乳酸菌与酵母菌之间可能存在相互作用的关系。      

自然发酵风干肠中乳酸菌的分离与鉴定

以自然发酵风干肠为研究对象,分析了细菌总数和乳酸菌菌数的变化情况,结果表明,乳酸菌为风干肠发酵过程中的优势菌群。通过对风干肠中乳酸菌的分离鉴定,共分离出戊糖片球菌(Pediococcus pentosaceus)、乳酸乳球菌乳酸亚种(Lactococcus lactis subsplactis)、短乳杆菌(Lactobacillus brevis)、弯曲乳杆菌(Lactobacillus curvatus)和发酵乳杆菌(Lactobacillus fermentum)5株乳酸菌。24h产酸速率测定结果表明,弯曲乳杆菌>短乳杆菌>乳酸乳球菌乳酸亚种>戊糖片球菌>发酵乳杆菌。      

Anti-Helicobacter pylori activity of fermented milk with lactic acid bacteria

BACKGROUND: Ten strains of lactic acid bacteria (LAB) were investigated for their anti‐Helicobacter pylori effects. The bactericidal activity and organic acid content in spent culture supernatants (SCS) from fermented milk were measured. In addition, the exclusion effect of SCS against H. pylori infection of human gastric epithelial AGS cells was assayed. RESULTS: Three LAB strains, LY1, LY5 and IF22, showed better anti‐Helicobacter effects than the other strains. There were no significant differences in the bactericidal activity of LAB strains between original SCS, artificial SCS and SCS treated by heating or protease digestion. However, neutralised SCS lost this activity. These results suggest that the anti‐H. pylori activity of SCS may be related to the concentration of organic acids and the pH value but not to protein components. In the AGS cell culture test, both fermented LY5‐SCS and artificial LY5‐SCS significantly reduced H. pylori infection and urease activity (P < 0.05). CONCLUSION: In this study, in vitro methods were used to screen potential probiotics with anti‐H. pylori activity. This may provide an excellent and rapid system for studying probiotics in the functional food and dairy industries. Copyright © 2011 Society of Chemical Industry     

发酵羊乳中乳酸菌低产粘菌株的筛选

对从商业乳酸菌发酵剂分离纯化的11株保加利亚乳杆菌和11株嗜热链球菌在发酵羊乳中的粘度进行了研究。结果表明,保加利亚乳杆菌L.b-124菌株和L.b-346菌株、嗜热链球菌S.t-222菌株和S.t-346菌株具有较低的产粘能力,凝乳时间也较短;将L.b-346菌株与S.t-346菌株按杆菌和球菌1:1比例混合后在43℃下发酵羊乳时,可获得较低的发酵粘度。