Growth and metabolism of selected strains of probiotic bacteria in milk

Growth and metabolism of five probiotic strains with well-documented health effects were studied in ultra-high temperature (UHT) treated milk, supplemented with 0.5% (w/v) tryptone or 0.75% (w/v) fructose. The probiotic strains were Lactobacillus acidophilus La5, Lb. acidophilus 1748, Lactobacillus rhamnosus GG, Lactobacillus reuteri SD 2112 and Bifidobacterium animalis BB12. Fermentation was followed for 72 h at 37 degrees C and the samples were analysed for pH, log cfu ml(-1), volatile compounds, organic acids and carbon dioxide. The strains reduced pH from 6.7 to between 3.9 and 4.4 after 24 h of incubation. All strains attained viable cell counts above 8.7-9.18 log cfu ml(-1) after 6-16 h of incubation. The two Lb. acidophilus strains showed a stable level of viable cells during 12-72 h of incubation but the three other strains showed a reduction of 0.4-1.1 log cfu ml(-1) from 24 to 72 h of incubation. However, all strains showed cell levels between 7.8 and 8.7 log cfu ml(-1) after 72 h of incubation. After 48 h of incubation, the amount of lactic acid produced varied according to strain from 6949 to 14,000 mg kg(-1) and acetic acid produced varied from 0 to 6901 mg kg(-1). Three of the strains metabolised citrate but only low amounts of diacetyl and acetoin were detected within strains, 0.2-0.8 and 6.5-10 mg kg(-1), respectively. Carbon dioxide produced varied from 221 to 3942 mg kg(-1) and was connected to the citrate-fermenting ability of the strain used and their carbohydrate fermentation pathway. Three of the strains produced detectable levels of acetaldehyde and the concentration varied from 9.4 to 12.6 mg kg(-1) after 24 h of incubation. All five probiotic strains showed very different profiles of metabolites during fermentation; however, the two Lb. acidophilus strains were the most alike. Our findings show the importance of controlling the fermentation time since the probiotic strains produced different amounts of metabolic products according to fermentation time.     

益生菌Lactobacillus casei Zhang发酵特性的评价

以L.acidophilus NCFM,L.rhamnosus GG,L.casei Shirota和Bifidobacterium animalis Bb12为参照,对益生菌L.casei Zhang的发酵特性进行了研究.结果表明:发酵结束时,添加L.casei Zhang的发酵乳具有较高的pH值和较低的滴定酸度.4℃贮藏28 d后,L.casei Shirota和Bifidobacterium animalis Bb12的蛋白水解能力下降,而L.asei Zhang,L addophilusNCFM和L.rhamnosus GG蛋白水解能力不断上升.尽管Lcasei Zhang在脱脂乳中的发酵速度低于对照益生菌.但贮藏期内其活菌数缓慢上升并于贮藏结束时达到最高.L.casei Zhang完全具备益生菌所应有的发酵特性,因此其适合应用于发酵乳的开发.     

Manufacture of Turkish Beyaz cheese added with probiotic strains

The survival of the probiotic strains Lactobacillus fermentum (AB5-18 and AK4-120) and Lactobacillus plantarum (AB16-65 and AC18-82), all derived from human faces, was investigated in Turkish Beyaz cheese production. Three batches of Turkish Beyaz cheese were produced: one with the test probiotic culture mix (P), another with a commercial starter culture mix including Lactoccocus lactis subsp. cremoris, Lactococcus lactis subsp. lactis (C) and the third with equal parts of the commercial starter culture mix and test probiotic culture mix (CP). The cheeses were ripened at 4 °C for 120 days and the viability of cultures was determined monthly. Cheese samples were analyzed for total solids, fat in solids, titratable acidity, pH, salt in total solids, proteolysis, sensory evaluation, aroma compounds and biogenic amines. While initial lactic acid bacteria load in P cheese was 2.7 × 10⁹ at the beginning, it was 7.42 × 10⁷ cfu/g at the end of 120 days of ripening. The results showed that test probiotic culture mix was successfully used in cheese production without adversely affecting the cheese quality during ripening. The chemical composition and sensory quality of P cheeses were also comparable with C cheeses. The present study indicates that probiotic cultures of human origin are feasible for Turkish Beyaz cheese production.     

In vitro comparison of probiotic properties of Lactobacillus casei Zhang, a potential new probiotic, with selected probiotic strains

Since 2004, our research group has isolated 240 Lactobacillus strains from Koumiss, a traditional fermented alcoholic beverage prepared from mare's milk in Inner Mongolia, Xinjiang of China and Mongolia. Among these Lactobacillus strains a novel strain with potential probiotic properties, Lactobacillus casei Zhang, was screened out and studied extensively for its probiotic properties, health-promoting effects and fermentation characteristics. In vitro tests indicated that L. casei Zhang had high tolerance to simulated gastric, intestine juices and bile salts, similar to commercial probiotic strains such as Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus GG, L. casei Shirota and Bifidobacterium animalis Bb12. Higher acid-production activity and proteolytic activity was observed in the fermented milk inoculated with L. casei Zhang during the refrigerated storage than in the samples inoculated with the selected commercial probiotics. The yogurt samples fermented with L. casei Zhang exhibited similarer viable count (1.0 × 10⁹ cfu/mL) as the other samples after 28 d of refrigerated storage. The results suggested that L. casei Zhang showed good potential for application in functional foods and health-related products.     

Texture profile of fermented rice extracts with probiotic strains and different contents of waxy maize starch,and sensory acceptance of flavoured selected extract

Rice is a staple food for a great part of the world’s population, and its processing generates a great volume of low value by-products, such as bran and broken grains. This work aimed to elaborate fermented rice by-products extracts with probiotic strains and with different waxy maize starch (WMS) contents (0%, 4%, 8%, 12% and 16% w/w), in order to select one with texture profile similar to Greek-style yogurt, and to characterise the chemical composition and sensory acceptance of the selected extract added with artificial strawberry aroma and fresh strawberry syrup. The texture profile of the extract changed with WMS content and the 4% WMS fermented rice by-products extract was the most similar to Greek-style yogurt. The flavoured fermented rice by-products with WMS product presented 72.67% moisture, 2.55% protein, 0.2% lipid, 0.8% ash (340 mg 100 g⁻¹ of potassium), 27.4 µmol Trolox g⁻¹ and 134.74 mg GAE 100 g⁻¹ of phenolic compounds. The flavoured fermented rice by-products extract did not show microbiological risk and presented probiotic characteristic. Regarding the sensory analysis, it was observed that the aroma stood out among the other attributes, obtaining the highest score and acceptance index, while colour, flavour and texture scored ‘liked regularly to moderately’. The flavoured fermented rice by-products extract is an innovative product and feasible regarding the technological, physical, chemical, microbiological and sensory characteristics, having great potential to be inserted into the market.     

益生乳杆菌的筛选研究

以分离保存的15株乳杆菌为研究对象,从中筛选益生乳杆菌两株。通过检测乳杆菌的疏水性和自聚性,得到表面疏水性和自聚性均较高的菌株为Ind-3、CH10和M8。其中,乳杆菌CH10和M8在模拟胃肠环境下活菌数均达到106mL-1以上。通过灌胃乳杆菌CH10和M8,表明小鼠体重增加明显高与对照组,且实验组小鼠粪便中的β-半乳糖苷酶活性明显高于对照组。灌胃两株乳杆菌后,小鼠肠道内双歧杆菌与乳杆菌活菌数提高了,且肠杆菌、肠球菌和产气荚膜梭菌的生长繁殖受到不同程度的抑制,而对照组无明显变化。结果表明筛选出的两株乳杆菌可以促进小鼠对营养物质的吸收,提高肠道内有益菌的数量,抑制有害菌的生长繁殖,或许能够改善寄主肠道的微生态环境。     

Incorporation of selected nutraceuticals and probiotic bacteria into a fermented milk

Yoghurts were produced from a base milk containing three important nutraceuticals, namely ω-3-fatty acids, isoflavones and phytosterols. The cultures employed to make the yoghurts were single probiotic strains of Lactobacillus gasseri or Bifidobacterium infantis and, to achieve a short production time, a two-stage fermentation procedure was used with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus providing the rapid acidification. Yoghurts containing counts of >1.0×10⁸ cfu mL⁻¹ of the individual probiotics and high counts of the traditional species from yoghurt were awarded overall scores for sensory acceptability >4.0 out of 5.0; the nutraceuticals appeared to have no adverse effect on flavour. Storage trials at 5 °C showed that the viability of the probiotic cultures was retained over 15 days.     

不同益生菌的加入对酸奶储藏期内风味的影响

以LGG、NCFM、BB-12和ABY四种益生菌酸奶为研究对象,跟踪检测这些样品在20℃储藏条件下的滴定酸度、益生菌活菌数、双乙酰含量和乙醛含量,比较不同益生菌对酸奶风味及酸度的影响。研究表明,在20℃常温储藏条件下,4种益生菌都不同程度地加速了酸奶的后酸化,其中以LGG和NCFM最为明显;除LGG外,NCFM,BB-12和ABY在酸奶储藏期内的存活率都呈下降趋势;不同益生菌酸奶中双乙酰含量和乙醛含量有明显差异,在添加LGG的酸奶中含量最低,在添加NCFM的酸奶中含量最高。     

N-nitrosamine content of New Zealand beer and malted barley

The nitrosamine content of 16 New Zealand beers and one Australian beer, all manufactured in 1980, was determined by chemiluminescence. The levels, calculated as N-nitrosodimethylamine (NDMA) ranged from 0.2–1.8 μg dm^?3 with a mean of 0.76 μg dm^?3. Over a period of 2 years, the NDMA levels in New Zealand malt have been reduced as the result of changes in kilning technology, introduced to control the level of nitrogen oxides in kiln air. For example, the mean value for directly-fired malt was 5.4 μg kg^?1, that for malt dried in air heated by burners to which sulphur had been added was 2.4 μg kg^?1 and that for malt dried in air to which SO₂ had been released was 0.3 μg kg^?1. For the purposes of quality control, screening beer or malt for nitrosamines can be accomplished adequately by the injection of CH₂Cl₂ extracts directly into a thermal energy analyser.     

具有抗氧化和抑菌能力的益生性乳酸菌筛选及鉴定

为筛选出具有抗氧化和抑菌能力的益生性乳酸菌菌株,对本实验室保存的18株乳酸菌进行耐酸耐胆盐、疏水性、自由基清除能力和抑菌能力分析,利用16S rDNA和管家基因pheS和rpoA序列同源性分析对筛选菌株进行鉴定。结果表明,菌株A1、A7、A13、A14、A15在pH 3.0时的存活率均大于70%,在牛胆盐质量分数为03%时的存活率均大于40%。菌株A15的产酸能力最强,24 h时的pH值为3.68。菌株A1、A7、A14、A15均具有高疏水性。菌株A15完整细胞悬液的羟自由基清除能力(39%)和DPPH自由基清除能力(51%)最强,菌株A14次之。此外,菌株A15对金黄色葡萄球菌和大肠杆菌的生长均有抑制作用。16S rDNA和管家基因pheS和rpoA序列同源性分析结果表明:A1为乳肠球菌、A7为副干酪乳杆菌、A14为副植物乳杆菌、A15为植物乳杆菌植物亚种,为开发新型营养健康乳酸菌发酵食品奠定基础。     

潜在益生菌菌株脱除胆固醇特性研究

采用气相色谱测定胆固醇的方法，对分离出来的7个潜在益生菌菌株进行了体外纯培养模式降低胆固醇的实验，结果表明，双歧杆菌普遍具有较强的降低胆固醇能力，其次是干酪乳杆菌。总之，进行功能性实验的7株益生菌在纯培养模型和发酵模型中均表现出了不同的脱除胆固醇能力，这主要是由菌株本身的特性决定的.     

Screening selected strains of probiotic lactic acid bacteria for their ability to produce biogenic amines (histamine and tyramine)

The aim of this study was to investigate the production of biogenic amines (BA), histamine and tyramine by some probiotic lactic acid bacteria (LAB). Fifteen strains representing six LAB species were screened qualitatively by growing them in a decarboxylase medium. Quantitative analysis was carried out by HPLC analysis with direct derivatization of acid extracts. Lactobacillus casei (TISTR 389) and Lactobacillus delbrueckii subsp. bulgaricus (TISTR 895) were found to produce BA. The highest levels of histamine (1820.9 ± 3.5 mg L^?1) and tyramine (5486.99 ± 47.6 mg L^?1) formation were observed for the TISTR 389 strain, while TISTR 895 produced only histamine (459.1 ± 0.63 mg L^?1) in the decarboxylase broth. Biogenic amine potential was not observed for the Lactobacillus acidophilus, Lactobacillus lactis subsp. lactis, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum strains studied. This study confirmed that BA formation is strain dependent and not related to the species. Therefore, careful screening for amino acid decarboxylase activity is recommended before selecting LAB as appropriate starter or probiotic strains in food and dairy industry.     

Growth of probiotic bacteria and bifidobacteria in a soy yogurt formulation

Soy beverage and cows' milk yogurts were produced with Steptococcus thermophilus (ATCC 4356) and Lactobacillus delbrueckii subsp. bulgaricus (IM 025). The drop in pH during fermentation was faster in the soy beverage than in cows' milk, but the final pH values were similar. Yogurts were prepared with a yogurt starter in conjunction with either the probiotic bacteria Lactobacillus johnsonii NCC533 (La-1), Lactobacillus rhamnosus ATCC 53103 (GG) or human derived bifidobacteria. The presence of the probiotic bacteria did not affect the growth of the yogurt strains. Approximately 2 log increases in both L. rhamnosus GG and L. johnsonii La-1 were observed when each was added with the yogurt strains in both cows' milk and the soy beverage. Two of the five bifidobacteria strains grew well in the cows' milk and soy beverage during fermentation with the yogurt bacteria. High pressure liquid chromatography (HPLC) analyses showed that the probiotic bacteria and the bifidobacteria were using different sugars to support their growth, depending on whether the bacteria were growing in cows' milk or soy beverage.     

Effect of temperature on growth and metabolism of probiotic bacteria in milk

The growth and metabolism of six probiotic strains with documented health effects were studied in ultra-high temperature (UHT) treated milk supplemented with 0.5% (w/v) tryptone or 0.75% (w/v) fructose at different temperatures. The probiotic strains were Lactobacillus acidophilus La5, Lb. acidophilus 1748, Lb. johnsonii LA1, Lb. rhamnosus GG, Lb. reuteri SD 2112 and Bifidobacterium animalis BB12. Fermentation was followed for 48 h at 20, 30, 37 and 45 °C and the samples were analysed for pH, log cfu mL⁻¹, volatile compounds, organic acids and carbon dioxide. All six probiotic strains showed very different profiles of metabolites during fermentation, however, the two Lb. acidophilus strains were the most alike. All strains, except Lb. reuteri SD 2112, showed viable cell numbers above 6.5 log cfu mL⁻¹ after 48 h fermentation at 30, 37 and 45 °C. The probiotic strains produced different amounts of metabolic products according to temperature and fermentation time illustrating the importance of controlling these parameters.     

Influence of probiotic strains added to cottage cheese on generation of potentially antioxidant peptides,anti-listerial activity,and survival of probiotic microorganisms in simulated gastrointestinal conditions

Our objective was to evaluate the viability of probiotic microorganisms added to cottage cheese under simulated gastrointestinal conditions, the release of potentially-antioxidant peptides, and their antimicrobial effect on Listeria monocytogenes. Cottage cheeses were prepared in triplicate, incorporating Lactobacillus casei, Lactobacillus rhamnosus GG, the commercial mix YO-MIX™ 205, or a control without probiotic addition. The probiotic population remained at >10⁶ cfu g⁻¹ during 28 days of storage at 8 °C. Cheeses made with added probiotics showed an increased metabolic activity with higher levels of lactic and acetic acids. Higher numbers of potentially bioactive peptides were observed in cheeses added with probiotics. L. monocytogenes population was reduced by about one log cycle after 20 days of storage, in cheeses with probiotics added. Our results indicate that cottage cheese is a good vehicle for probiotic bacteria.     

Antimicrobial effects of probiotic bacteria against selected species of yeasts and moulds in cheese-based dips

The antimicrobial properties of selected probiotic bacteria against Aspergillus niger , Penicillium roqueforti , Fusarium spp., Candida albicans and Saccharomyces cerevisiae were examined. Well diffusion and spot and streak methods showed strong inhibition effect of probiotic bacteria and their metabolites against moulds and minimal effect against yeasts. Among the moulds species tested, the inhibitory effect was strongest against Fusarium spp., moderate against Penicillium roqueforti and minimal against A. niger . All strains of Lactobacillus rhamnosus and L. paracasei subsp. paracasei showed maximum inhibitory effect. When probiotic bacteria and yeasts and moulds were co-cultured in broth media, strains of L. rhamnosus showed maximum inhibitory effect, whereas L. paracasei subsp. paracasei , L. acidophilus , Bifidobacterium animalis and Propionibacterium showed moderate inhibitory effect against C. albicans . Saccharomyces cerevisiae was minimally controlled by probiotic bacteria. Pre-grown probiotic bacterial culture metabolites controlled yeasts and moulds more effectively than their freeze-dried or frozen forms. Adding metabolites of probiotic bacteria (5% w/w) showed an effective control against A. niger , Fusarium spp. and C. albicans during the shelf life of 10 weeks at 4 °C and no colonies of yeasts and moulds were formed on the surface of the dip.     

African maize porridge: a food with slow in vitro starch digestibility

The incidence of diabetes is very low in rural, traditionally living, South African Black people, but higher in the urbanised Black population. One factor, that could have contributed to the increased prevalence of diabetes, is the change in diet from maize porridge to bread. An in vitro method was used to determine the starch digestibility of African maize porridge compared to other cereal foods. Maize porridge had a much lower in vitro starch digestibility than white bread (P<0.001). There was a positive correlation (P=0.05) between rate of starch digestibility of maize porridge and endosperm hardness. Decreasing the particle size of the maize meal by conversion to maize flour did not increase starch digestibility. Both decreasing and increasing the cooking time decreased the starch digestibility. The predicted glycaemic index for maize porridge ranged from 39 to 50 (glucose standard), which suggests that maize porridge may be useful in the dietary management of diabetes.     

Influence of malt,wheat, and barley extracts on the bile tolerance of selected strains of lactobacilli

The bile tolerance of three strains of lactobacilli, selected on the basis of their ability to ferment cereal substrates and tolerate simulated gastric transit was examined. Lactobacillus reuteri (NCIMB 1195), L. acidophilus (NCIMB 8821), and L. plantarum (NCIMB 8826) were exposed to 2% bile for 4 h in a phosphate-saline buffer at pH 7. The effects of supplementing the assays with cereal extracts, increasing concentrations of glucose, and free amino nitrogen was also examined. In the absence of any supplements L. reuteri showed the greatest resistance to bile, whilst L. acidophilus displayed the greatest sensitivity. Addition of cereal extracts improved the tolerance of all three strains, barley and wheat extracts imparted similar levels of enhancement in viability, whilst the addition of malt showed a greater positive influence. The extent of improvement was related to the concentration of soluble sugars and FAN present in the cereals, this was illustrated further by the addition of glucose to the assays, as the viability of the organisms was progressively improved with increasing concentrations. The presence of free amino nitrogen also increased bacterial resistance to bile but to a lesser extent both glucose and cereal extracts.