Evolution of carbohydrate fraction in carbonated fermented milks as affected by beta-galactosidase activity of starter strains

The influence of carbonation on the evolution of lactose, galactose and glucose in fermented milks with added probiotic bacteria (Lactobacillus casei, Lactobacillus acidophilus and/or Bifidobacterium bifidum) was evaluated and related to beta-galactosidase activity of starter strains. During incubation and first days of refrigeration, lactose hydrolysis resulting in the liberation of galactose and glucose occurred in CT (Streptococcus thermophilus/Lb. casei), AT (Str. thermophilus/Lb. acidophilus) and ABT fermented milks (Str. thermophilus/Lb. acidophilus/Bifid. bifidum). Levels of galactose were higher than those of glucose and could be related to the preferential consumption of glucose by actively growing bacteria. Through the incubation, lactose and monosaccharide levels were not affected by milk carbonation. However, during refrigerated storage the presence of this gas was associated with slightly lower content of lactose and higher levels of galactose and glucose in AT and ABT products but not in CT fermented milks. Through the refrigeration galactose was moderately utilised by Lb. acidophilus in AT products whereas the presence of Bifid. bifidum seems to prevent the consumption of this sugar in ABT fermented milks. Glucose remained constant, with minor variations in CT products but a continuous increase of this sugar occurred in carbonated AT and ABT fermented milks during storage. Beta-galactosidase activity displayed by Str. thermophilus strains was similar at pH 6.5 (initial pH of non-carbonated samples) and pH 6.3 (initial pH of carbonated samples) whereas Lb. acidophilus LaA3 showed greater beta-galactosidase activity at pH 6.3 than at higher pH values. Thus, the enhanced metabolic activity of Lb. acidophilus caused by the low initial pH of carbonated milk also promoted higher cellular beta-galactosidase activity that could have released greater amounts of galactose and glucose from lactose in AT and ABT fermented milks through the refrigerated period. In CT fermented milks, similar beta-galactosidase activity levels of Str. thermophilus at pH 6.5 and 6.3 together with the absence of beta-galactosidase activity in Lb. casei could explain the lack of differences on glucose and galactose content between carbonated and non-carbonated samples.     

Viability of probiotic (Bifidobacterium, Lactobacillus acidophilus and Lactobacillus casei) and nonprobiotic microflora in Argentinian Fresco cheese

We evaluated the suitability of Argentinian Fresco cheese as a food carrier of probiotic cultures. We used cultures of Bifidobacterium bifidum (two strains), Bifidobacterium longum (two strains), Bifidobacterium sp. (one strain), Lactobacillus acidophilus (two strains), and Lactobacillus casei (two strains) in different combinations, as probiotic adjuncts. Probiotic, lactic starter (Lactococcus lactis and Streptococcus thermophilus), and contaminant (coliforms, yeasts, and molds) organisms were counted at 0, 30, and 60 d of refrigerated storage. Furthermore, the acid resistance of probiotic and starter bacteria was determined from hydrochloric solutions (pH 2 and 3) of Fresco cheese. The results showed that nine different combinations of bifidobacteria and L. acidophilus had a satisfactory viability (count decreases in 60 d <1 log order) in the cheese. Both combinations of bifidobacteria and L. casei cultures assayed also showed a satisfactory survival (counts decreased <1 log order for bifidobacteria but no decrease was detected for L. casei). On the other hand, the three combinations of bifidobacteria, L. acidophilus, and L. casei tested adapted well to the Fresco cheese environment. When a cheese homogenate at pH 3 was used to partially simulate the acidic conditions in the stomach, the probiotic cultures had an excellent ability to remain viable up to 3 h. At pH 2, the cell viability was more affected; B. bifidum was the most resistant organism. This study showed that the Argentinian Fresco cheese could be used as an adequate carrier of probiotic bacteria.     

Antiviral activities of cell-free supernatants of yogurts metabolites against some RNA viruses

The antiviral activity of the cell-free supernatants (CFS) containing the metabolites of five yogurts fermented under anaerobic conditions with Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantarum, Streptococcus thermophilus and Bifidobacterium bifidum, respectively, was initially evaluated against seven RNA viruses using virus-induced cytopathic effects reduction method, while comparing that of De Man, Rogosa, and Sharpe (MRS) broths containing metabolites fermented with the same bacteria. All the CFS of yogurt showed high antiviral activity against seven RNA viruses, while the CFS of MRS broth exhibited similar or lower antiviral activity. Each CFS of yogurt and MRS broth showed no cytotoxicity with normal morphology in Vero or MDCK cells. Furthermore, CFS of each yogurt was more effective than that of MRS broth against the three enteroviruses and CFS of yogurt containing metabolites fermented with L. plantarum exhibited strong anti-influenza virus activity among that of the other yogurt and MRS broth. Therefore, CFS of the yogurt containing metabolites fermented with probiotic bacteria showed high potential to be used for developing fermented milk-based foods or drugs.     

Prevention of gastrointestinal infection using immunobiological methods with milk fermented with Lactobacillus casei and Lactobacillus acidophilus

The protective effect of feeding milk fermented with a mixture of Lactobacillus casei sp. and Lb. acidophilus sp. against Salmonella typhimurium infection in mice was compared with that obtained feeding milks fermented with these microorganisms individually. The survival rate obtained after oral infection with Sal. typhimurium was 100% in mice pretreated by feeding during 8 d with the mixture of Lb. casei and Lb. acidophilus fermented milks. Similar treatments with the individual milks were ineffective. Moreover, mice became more susceptible to infection with Sal. typhimurium after such treatment. The colonization of liver and spleen with the pathogen was markedly inhibited by the pretreatment with the mixture of fermented milk, while such inhibition was not observed using the Lb. casei and Lb. acidophilus milks. The highest levels of anti-salmonellae antibodies in serum and in intestinal fluid were found in the group of mice fed with the mixture and with Lb. casei fermented milk respectively. However, this latter milk was not effective in protecting against Sal. typhimurium. When the mice were first infected with Sal. typhimurium and then fed with the mixture of fermented milks, pathogen colonization was not prevented. The results suggest that the augmentation of resistance to salmonellae caused by the treatment with Lb. casei- + Lb. acidophilus-fermented milk was due to the anti-salmonellae protective immunity mainly mediated by the mucosal tissue. Milk fermented with this mixture could be used as an immunobiological method to prevent gastrointestinal infection.     

Investigation of the possible use of probiotics in ice cream manufacture

This study attempted to investigate the possibility of using some types of probiotic bacteria in the production of ice cream and was aimed at making a contribution to the manufacture of new functional foods. For this purpose, different cream levels (5% and 10%) and different strains of probiotic bacteria ( Lactobacillus acidophilus , Bifidobacterium bifidum and both) were used in ice cream production to determine their effects on the quality of the ice creams in each group. During storage of 1, 15, 30, 45, 60, 75 and 90 days, L. acidophilus , B. bifidum counts and sensory analyses were performed. The results obtained at the end of storage demonstrated that the counts of L. acidophilus and B. bifidum continued to decrease during the storage but all types of ice cream sample seemed to preserve their probiotic property even after 90 days. Higher counts of probiotic bacteria were observed in the samples with L. acidophilus , B. bifidum , and L. acidophilus and B. bifidum in double-cultured samples respectively. In general, it appeared that the ice cream samples with 5% cream content were found to be more delectable. All ice cream samples have shown good results in preserving their probiotical properties for more than 3 months. Although sensory scores of probiotic ice cream samples reduced during this time, they rated as 'tasty' throughout the storage.     

Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in ice cream for use as a probiotic food.

Probiotic ice cream was made by fermenting a standard ice cream mix with Lactobacillus acidophilus and Bifidobacterium bifidum cultures and then freezing the mix in a batch freezer. Survival of the L. acidophilus and B. bifidum, as well as beta-galactosidase activity, was monitored during 17 wk of frozen storage at -29 degrees C. After freezing of the fermented mix, bacterial counts were 1.5 x 10(8) cfu/ml for L. acidophilus and 2.5 x 10(8) cfu/ml for B. bifidum. Seventeen weeks after freezing, these counts had decreased to 4 x 10(6) and 1 x 10(7) cfu/ml, respectively. During the same period, beta-galactosidase activity decreased from 1800 to 1300 units/ml. Probiotic ice cream was prepared at pH 5.0, 5.5, and 6.0 to determine consumer preferences and was compared with standard Utah State University "Aggie" ice cream. All samples were strawberry-flavored and were evaluated by 88 judges. The preferred pH of probiotic ice cream, based on overall acceptance, was pH 5.5. We demonstrated that probiotic ice cream is a suitable vehicle for delivering beneficial microorganisms such as L. acidophilus and B. bifidum to consumers. The bacteria can be grown to high numbers in ice cream mix and remain viable during frozen storage.     

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.     

嗜酸乳杆菌、双歧杆菌和干酪乳杆菌的选择性计数

介绍了干酪乳杆菌、嗜酸乳杆菌和双歧杆菌的选择性计数方法。LC培养基只能计数干酪乳杆菌,MRS-水杨素（或山梨醇）培养基可以计数嗜酸乳杆菌和干酪乳杆菌;而MRS培养基可以计数这3种益生菌,通过减法原则从嗜酸乳杆菌、干酪乳杆菌和双歧杆菌混合物中单独计数。另外,MRS-NNLP培养基也可用于选择性计数双歧杆菌。     

两歧双歧杆菌与嗜酸乳杆菌微生态制剂的工业化生产工艺研究

研究了两歧双歧杆菌和嗜酸乳杆菌双联微生态制剂的工业化生产工艺。对发酵培养基及发酵条件进行了研究和优化，并确定了发酵奶的冷冻干燥工艺条件。冻干后的产品中两歧双歧杆菌和嗜酸乳杆菌的活菌数均达到了10^9-10^10g^-1，而且在冻干工艺条件下，发酵产物中的各种活性成分得以最大限度地保存下来，使产品可以达到作为微生态制剂应用的相关标准。     

Acid, bile, and heat tolerance of free and microencapsulated probiotic bacteria

ABSTRACT:  Eight strains of probiotic bacteria, including Lactobacillus rhamnosus , Bifidobacterium longum, L. salivarius, L. plantarum , L. acidophilus , L. paracasei , B. lactis type Bl-O4, and B. lactis type Bi-07, were studied for their acid, bile, and heat tolerance. Microencapsulation in alginate matrix was used to enhance survival of the bacteria in acid and bile as well as a brief exposure to heat. Free probiotic organisms were used as a control. The acid tolerance of probiotic organisms was tested using HCl in MRS broth over a 2-h incubation period. Bile tolerance was tested using 2 types of bile salts, oxgall and taurocholic acid, over an 8-h incubation period. Heat tolerance was tested by exposing the probiotic organisms to 65 °C for up to 1 h. Results indicated microencapsulated probiotic bacteria survived better ( P < 0.05) than free probiotic bacteria in MRS containing HCl. When free probiotic bacteria were exposed to oxgall, viability was reduced by 6.51-log CFU/mL, whereas only 3.36-log CFU/mL was lost in microencapsulated strains. At 30 min of heat treatment, microencapsulated probiotic bacteria survived with an average loss of only 4.17-log CFU/mL, compared to 6.74-log CFU/mL loss with free probiotic bacteria. However, after 1 h of heating both free and microencapsulated probiotic strains showed similar losses in viability. Overall microencapsulation improved the survival of probiotic bacteria when exposed to acidic conditions, bile salts, and mild heat treatment.     

Fibers from fruit by-products enhance probiotic viability and fatty acid profile and increase CLA content in yoghurts

This study evaluated the effect of the supplementation of total dietary fiber from apple, banana or passion fruit processing by-products on the post-acidification, total titratable acidity, bacteria counts and fatty acid profiles in skim milk yoghurts co-fermented by four different probiotics strains: Lactobacillus acidophilus L10 and Bifidobacterium animalis subsp. lactis BL04, HN019 and B94. Apple and banana fibers increased the probiotic viability during shelf-life. All the fibers were able to increase the short chain and polyunsaturated fatty acid contents of yoghurts compared to their respective controls. A synergistic effect between the type of fiber and the probiotic strain on the conjugated linoleic acid content was observed, and the amount of α-linolenic acid was increased by banana fiber. The results of this study demonstrate, for the first time, that fruit fibers can improve the fatty acid profile of probiotic yoghurts and point out the suitability of using fibers from fruit processing the by-products to develop new high value-added fermented dairy products.     

PCR method for detection and identification of Lactobacillus casei/paracasei bacteriophages in dairy products

Bacteriophage infections of starter lactic acid bacteria (LAB) pose a serious risk to the dairy industry. Nowadays, the expanding use of valuable Lactobacillus strains as probiotic starters determines an increase in the frequency of specific bacteriophage infections in dairy plants. This work describes a simple and rapid Polymerase Chain Reaction (PCR) method that detects and identifies bacteriophages infecting Lactobacillus casei/paracasei, the main bacterial species used as probiotic. Based on a highly conserved region of the NTP-binding genes belonging to the replication module of L. casei phages phiA2 and phiAT3 (the only two whose genomes are completely sequenced), a pair of primers was designed to generate a specific fragment. Furthermore, this PCR detection method proved to be a useful tool for monitoring and identifying L. casei/paracasei phages in industrial samples since specific PCR signals were obtained from phage contaminated milk (detection limit: 10(4) PFU/mL milk) and other commercial samples (fermented milks and cheese whey) that include L. casei/paracasei as probiotic starter (detection limit: 10(6) PFU/mL fermented milk). Since this method can detect the above phages in industrial samples and can be easily incorporated into dairy industry routines, it might be readily used to earmark contaminated milk for use in processes that do not involve susceptible starter organisms, or processes which involve phage-deactivating conditions.     

益生菌对发酵菠萝果汁综合品质的影响

菠萝果汁分别接种干酪乳杆菌、植物乳杆菌、嗜酸乳杆菌和肠膜明串珠菌进行发酵,研究菠萝果汁发酵过程中理化品质和感官品质的变化规律。结果表明,4种乳酸菌在菠萝果汁中生长良好,发酵24 h,植物乳杆菌和肠膜明串珠菌的活菌数对数值均已超过8.3;经乳酸菌发酵后,菠萝果汁的pH、维生素C（VC）含量均显著降低,而总酸、可溶性固形物、总酚、1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率显著升高。其中,植物乳杆菌具有较强的产酸能力,肠膜明串珠菌能够延缓VC的流失。在对感官品质影响方面,4种发酵菠萝果汁之间存在差异,经肠膜明串株菌发酵的菠萝果汁感官评价较优（79.79分）,而经嗜酸乳杆菌能够更好地保留菠萝果汁特有的挥发性香气成分。研究结果为益生菌菠萝果汁产品的开发提供参考依据和技术支持。     

Starter cultures employed in the manufacture of biofermented milks

The market for fermented milks is expanding as more consumers become interested in the possible health aspects of the product. This paper reviews the types of bacteria that are currently being used in the manufacture of biofermented milks. The requirements of a starter culture in terms of ability to ferment milk to a stable product and to provide the necessary sensory properties for consumer demands are outlined. The paper also discusses and highlights some of the problems encountered in developing milks for the health market using probiotic organisms such as Lactobacillus acidophilus and species of Bifidobacterium.     

On the viability of five probiotic strains when immobilised on various polymers

The viability of probiotic bacteria in six food‐grade polymers, at two concentrations, was evaluated in order to predict their feasibility as materials for bacterium immobilisation. Alginate and whey proteins were the most adequate polymers, except for Lactobacillus acidophilus Ki and Lactobacillus casei 01 at 2% (m/v) alginate. Xanthan gum appeared to be a potential vector for three strains. L‐carrageenan was adequate for both Bifidobacterium strains, but not for Lactobacillus at 2% (m/v). Bifidobacterium strains were not negatively affected by cellulose acetate phthalate, while the opposite held for L. acidophilus strains. Chitosan was the poorest polymer for immobilisation of probiotic bacteria.     

Isolation and identification of lactobacilli from novel-type probiotic and mild yoghurts and their stability during refrigerated storage.

A total of 26 Lactobacillus strains were isolated from various mild yoghurts and novel-type probiotic dairy products and from a starter culture preparation and were identified by using DNA-DNA hybridization technique. The species present in those products were found to be Lactobacillus acidophilus, L. johnsonii, L. crispatus, L. casei, L. paracasei and L. rhamnosus. DNA homology analysis revealed that some strains had been misclassified by their investigators. Three strains designated as L. acidophilus (L. acidophilus LA-1, L. acidophilus ATCC 43121 and the Lactobacillus strain from Biogarde culture) were found to belong to L. johnsonii and L. acidophilus L1 to be L. crispatus. Strains designated as L. casei were found to be members of three separate species: L. casei, L. paracasei and L. rhamnosus. Viable numbers of lactobacilli in mild and probiotic yoghurts varied greatly including some products with very low Lactobacillus counts. The majority of the probiotic yoghurts, however, contained viable counts above 10(5) per g even at the end of the best before use period.     

酵母菌乳酸菌共发酵对荔枝汁品质的影响

本研究以鲜榨荔枝汁为原料,分别对其进行干酪乳杆菌及三种酵母菌(BO213、D254、EC1118)的复合发酵,探索发酵过程中乳酸菌活菌数、pH、可溶性固形物、糖组分、有机酸、DPPH?清除能力以及感官指标等变化。结果表明：发酵后荔枝汁中的乳酸菌活菌数均高于7 lg (cfu/mL),均具有一定的益生功能。复合发酵不仅能促进干酪乳杆菌的生长,还能降低荔枝汁的糖度,且酒精度低于0.50%,符合无醇饮料的标准。随着发酵时间的延长,各组的糖含量均呈现下降的趋势。发酵24 h后干酪乳杆菌单独发酵组的乳酸含量显著低于复合发酵组,从高到低依次为：D254+干酪组(6.62 g/L)>Ec1118+干酪组(6.55 g/L)>BO213+干酪组(2.35 g/L)>干酪组(0.94 g/L)。D254+干酪组、Ec1118+干酪组发酵24 h的荔枝汁体外抗氧化能力(DPPH?清除能力)相比单独发酵组更具优势。发酵后的荔枝汁香气更佳,且复合发酵优于单独发酵,其中BO213+干酪组的香气最好,Ec1118+干酪组次之。综上,通过Ec1118+干酪乳杆菌可制得一款风味良好、营养丰富的发酵无醇益生荔枝饮料。     

Effect of milk base and starter culture on acidification,texture, and probiotic cell counts in fermented milk processing

In the present work, the compared effect of milk base and starter culture on acidification, texture, growth, and stability of probiotic bacteria in fermented milk processing, was studied. Two strains of probiotic bacteria were used, Lactobacillus acidophilus LA5 and L. rhamnosus LR35, with two starter cultures. One starter culture consisted only of Streptococcus thermophilus ST7 (single starter culture); the other was a yogurt mixed culture with S. thermophilus ST7 and L. bulgaricus LB12 (mixed starter culture). For the milk base preparation, four commercial dairy ingredients were tested (two milk protein concentrates and two casein hydrolysates). The resulting fermented milks were compared to those obtained with control milk (without enrichment) and milk added with skim milk powder. The performance of the two probiotic strains were opposite. L. acidophilus LA5 grew well on milk but showed a poor stability during storage. L. rhamnosus LR35 grew weakly on milk but was remarkably stable during storage. With the strains tested in this study, the use of the single starter culture and the addition of casein hydrolysate gave the best probiotic cell counts. The fermentation time was of about 11 h, and the probiotic level after five weeks of storage was greater than 106 cfu/ml for L. acidophilus LA5 and 10(7) cfu/ml for L. rhamnosus LR35. However, an optimization of the level of casein hydrolysate added to milk base has to be done, in order to improve texture and flavor when using this dairy ingredient.     

Production of beta-glucosidase and hydrolysis of isoflavone phytoestrogens by Lactobacillus acidophilus, Bifidobacterium lactis, and Lactobacillus casei in soymilk

ABSTRACT:  The study determined β-glucosidase activity of commercial probiotic organisms for hydrolysis of isoflavone to aglycones in fermenting soymilk. Soymilk made with soy protein isolate (SPI) was fermented with Lactobacillus acidophilus LAFTI^® L10, Bifidobacterium lactis LAFTI^® B94, and Lactobacillus casei LAFTI^® L26 at 37 °C for 48 h and the fermented soymilk was stored for 28 d at 4 °C. β-Glucosidase activity of organisms was determined using ρ-nitrophenyl β-D-glucopyranoside as a substrate and the hydrolysis of isoflavone glycosides to aglycones by these organisms was carried out. The highest level of growth occurred at 12 h for L. casei L26, 24 h for B. lactis B94, and 36 h for L. acidophilus L10 during fermentation in soymilk. Survival after storage at 4 °C for 28 d was 20%, 15%, and 11% greater ( P < 0.05) than initial cell counts, respectively. All the bacteria produced β-glucosidase, which hydrolyzed isoflavone β-glycosides to isoflavone aglycones. The decrease in the concentration of β-glycosides and the increase in the concentration of aglycones were significant ( P < 0.05) in the fermented soymilk. Increased isoflavone aglycone content in fermented soymilk is likely to improve the biological functionality of soymilk.     

含金属硫蛋白鸡蛋水解物对两种乳酸菌体外生长的影响

以乳酸菌生长过程中的pH、OD值和菌落总数为评价指标,研究了普通鸡蛋、含金属硫蛋白鸡蛋、普通鸡蛋水解物以及含金属硫蛋白的鸡蛋水解物对动物双歧杆菌、嗜酸乳杆菌以及两种混合菌的影响。结果表明,鸡蛋水解物作为氮源能够促进乳酸菌的增殖,经比较,最佳氮源为含金属硫蛋白鸡蛋水解物,其动物双歧杆菌、嗜酸乳杆菌和混合菌发酵液OD值为普通鸡蛋水解物的1.05倍、1.08倍和1.09倍。含金属硫蛋白的鸡蛋水解物在动物双歧杆菌、嗜酸乳杆菌以及两种混合菌发酵液的最佳添加量分别为2.0%、1.5%和2.0%,活菌数分别可达7.5×109、3.21×109和2.33×1010 CFU/mL。本研究为开发乳酸菌发酵金属硫蛋白鸡蛋肽产品提供指导。