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.     

Probiotic stability of yoghurts containing Jerusalem artichoke inulins during refrigerated storage

Experimentally prepared Jerusalem artichoke inulins (JAI) were compared with two commercial chicory root inulins for their prebiotic potentials in media broth model and growth-sustaining ability in non-fat yoghurts. Experimental yoghurts were made with 12% reconstituted skim milk (RSM) supplemented with 4% inulin powders, inoculated with mixed cultures of Lactobacillus casei LC-01, Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (1:0.5:0.5 based on supplier’s recommendation) and incubated overnight at 37 °C. Non-supplemented yoghurt was prepared from 16% RSM and used as control. The survival and acidifying activity of lactic and probiotic cultures in all yoghurts were investigated on weekly intervals during the shelf life of 28 days at 4 °C. Incorporation of JAI resulted in improved viability of LC-01, maintaining >7.0 log CFU/g during cold storage but did not affect the viability of yoghurt bacteria in comparison with the control.     

Survival of undesirable micro-organisms in fruit yoghurts during storage at different temperatures

The survival of undesirable micro-organismsYersinia enterocolitica,Escherichia coli,Rhodotorula mucilaginosa,Kluyveromyces marxianusin fruit yoghurts after inoculation at two different levels (10²–10³cfu/ml and 10⁴–10⁶cfu/ml) was investigated during storage at 4 and 8°C. When stored at 4°C, the survival ofY. enterocoliticawas not significant (3 days) except when the size of the initial inoculum was larger than 10⁵cfu/ml: in this case, viable cells of the pathogen were still recovered after 17 days of storage. Cells ofE. coliwere detected in the yoghurt samples after 24–48 days at 4°C, regardless of the inoculum size. The growth of yeasts easily occurred at 4°C;R. mucilaginosashowed an increase of 10–100 fold andK. marxianus) was able to grow from 10²to 10⁵–10⁶cfu/ml during the 30 days of storage. Similar trends were noted with 8°C storage. The survival of both contaminants was longer than 8–10 days and yeasts were able to grow remarkably well, with microbial values of 10⁶cfu/mlR. mucilaginosaand 10⁸cfu/ml (K. marxianusin the case of the large inoculum. Throughout the whole experiment, the counts for lactic streptococci were around 10⁸–10⁹cfu/ml whereas lactobacilli levels were 10⁶cfu/ml, and viability declined after 15 days of storage.The competitionin vitrobetween bothLactobacillusandStreptococcusstrains isolated from yoghurt andE. coliwas also investigated. WithLactobacillus, the inhibition ofE. coliwas very effective; no viableE. colicells were detected after 24 h incubation. TheStreptococcusstrain was also able to inhibitE. colibut not so rapidly; slight growth ofE. coliwas observed during the first 3 h, followed by a marked decrease in the number of cells.     

冷藏中低聚果糖对发酵乳中微生物的影响

对添加低聚果糖的原料奶，使用嗜酸乳杆菌、双歧杆菌和嗜热链球菌作为快速发酵剂进行发酵后．在4℃条件下储存42d。每周对发酵乳进行微生物分析和酸度测试。结果表明，低聚果糖质量浓度在10—50g／L时对嗜酸乳杆菌和嗜热链球菌的存活都没有太大影响。双歧杆菌活菌数下降幅度比嗜酸乳酸杆菌和嗜热链球菌要快。但添加低聚果糖对双歧杆菌的存活起到重要作用。结论是低聚果糖产品对双歧杆菌有益生作用，因此在冷藏发酵乳制品中有助于提高双歧杆菌活菌数。     

Influence of addition of raffinose family oligosaccharides on probiotic survival in fermented milk during refrigerated storage

The influence of the addition of raffinose family oligosaccharides (RFOs) extracted from lupin seeds on the survival of Bifidobacterium lactis Bb-12 and Lactobacillus acidophilus La-5 in fermented milk during 21 days of storage in refrigerated conditions was studied. For this purpose, viability and metabolic activity (expressed as pH, lactic and acetic acid production and utilization of soluble carbohydrates) of probiotic bacteria were determined. Retention of viability of B. lactis Bb-12 and L. acidophilus La-5 was greater in fermented milk with RFOs. The pH of probiotic fermented milk at 21 days of storage was lower (4.27) compared with probiotic fermented milk with RFOs (4.37). The highest levels of lactic and acetic acid were produced in probiotic fermented milk without RFOs compared with probiotic fermented milk with RFOs during storage at 4 °C. Soluble carbohydrates were utilised in fermented milk with and without RFOs, respectively, for maintaining B. lactis Bb-12 and L. acidophilus populations during refrigerated storage. In conclusion, all these experiments provide convincing evidence that RFOs have beneficial effects on the survival of these probiotic cultures in dairy products. As a result, such stored dairy products containing both probiotics and prebiotics have synergistic actions in the promotion of health.     

Effect of refrigerated storage time on the viability of probiotic bacteria in fermented probiotic milk drinks

The viability of Lactobacillus acidophilus and Bifidobacterium lactis in fermented milks A, B and AB, containing respectively these strains separately and mixed, was studied over 21 days of storage at 5°C. Samples were analysed for titratable acidity and viable cell counts of probiotics. Milks A and AB showed post-acidification mainly due to the β-galactosidase activity of L. acidophilus . Generally, the viability of the probiotics decreased during storage, whereas, in some cases, the proteoletic activity of L. acidophilus resulted in a higher survival rate of the probiotics even with the adverse effects of low temperature and organic acids.     

Survival and activity of selected probiotic organisms in set-type yoghurt during cold storage

The growth and metabolism of two probiotic organisms (L. acidophilus LAFTI^® L10 and Lactobacillus casei LAFTI^® L26) and a regular yoghurt culture (L. delbrueckii ssp. bulgaricus Lb1466 and Streptococcus thermophilus St1342) were studied in yoghurt containing 0.5%, 1.0%, and 1.5% (w/v) of high amylose corn starch powder (Hi-maize^®) or inulin. Viable cell counts of probiotic organisms, their metabolites and proteolytic activities, and viscosity of the yoghurts were determined during refrigerated storage for 28 d at 4 ^oC. In the presence of inulin, cultures showed better retention of viability (8.0 log cfu g⁻¹) in comparison with that of Hi-maize, which had a reduction by one log cycle. Lower concentrations of 0.5–1.0% Hi-maize improved (P<0.05) the production of propionic acid and also increased proteolytic activity of probiotic organisms substantially. A greater release of free amino acids may have sustained better growth of the organisms in yoghurts. Supplementation with either Hi-maize or inulin increased the viscosity of probiotic yoghurts significantly (P<0.05).     

Survival of human derived Lactobacillus plantarum in fermented cereal extracts during refrigerated storage

The aim of this study was to investigate the survival of a potentially probiotic Lactobacillus plantarum strain in barley, wheat and barley malt extracts. The extracts were produced from three flour/water suspensions, i.e., 5%, 20%, 30% w/w. After inoculation, the cultures were incubated for 24 h at 37 °C, and were subsequently stored at 4 °C for up to seventy days. The lactic acid and reducing sugar concentrations at the beginning of storage were significantly different between the fermented media, ranging from 0.5 g/L to 17 g/L and from 0.8 g/L to 6.5 g/L respectively, while the pH ranged between 2.9 and 3.4. It was observed that the cells survived much better in the malt extracts compared to barley and wheat extracts during refrigerated storage. Based on the results from a study using model media and supplemented cereal extracts it was derived that this was most likely due to their higher sugar concentration and the presence of protective unidentified compounds, albeit the fact that the malt extracts contained higher amounts of lactic acid.     

Effect of refrigerated storage temperature on the viability of probiotic micro-organisms in yogurt

The effect of refrigerated storage temperature was studied at 2, 5 and 8°C on the viability of probiotics in ABY ( Lactobacillus acidophilus, Bifidobacterium lactis BB-12 and yogurt bacteria. Bulgaricus , i.e. Streptococcus thermophilus and Lactobacillus delbrueckii ssp. Bulgaricus ) probiotic yogurt. The study was carried out during a 20-day refrigerated storage period to identify the best storage temperature(s). Also, the viability change of the probiotic micro-organisms was analysed at 5-day intervals throughout the refrigerated storage period. After 20 days, storage at 2°C resulted in the highest viability of L. acidophilus , whereas for Bifidobacterium lactis the highest viability was obtained when yogurt was stored at 8°C.     

肉鸡屠宰加工及冷藏中的微生物污染来源及菌相分析

研究了商业屠宰及冷藏过程对肉鸡胴体天然茵落的影响,结果表明,空气、预冷水与环境中各种接触面均是禽类生鲜产品的潜在污染源.尽管加工过程能减少胴体表面各种腐败茵、致病茵污染,但某些工序也增加了特定细菌的污染程度,细菌交叉污染发生在加工过程中的所有阶段.总的说来净膛操作会导致胴体污染程度上升,冲洗、预冷过程对减菌都有很好效果.但预冷过程也可能导致嗜冷茵污染的发生,并且加工过程中存活的细菌能在冷藏阶段继续增殖进而导致产品腐败.     

Short communication: Survival of the characteristic microbiota in probiotic fermented camel,cow, goat,and sheep milks during refrigerated storage

The objective of this study was to monitor the viability during storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in probiotic cultured dairy foods made from pasteurized camel, cow, goat, and sheep milks fermented by an ABT-type culture. The products manufactured were stored at 4°C for 42 d. Microbiological analyses were performed at weekly intervals. Streptococcus thermophilus CHCC 742/2130 was the most numerous culture component in all 4 products both at the beginning and at the end of storage. The viable counts of streptococci showed no significant decline in fermented camel milk throughout the entire storage period. The initial numbers of Lb. acidophilus LA-5 were over 2 orders of magnitude lower than those of Strep. thermophilus CHCC 742/2130. With the progress of time, a slow and constant decrease was observed in lactobacilli counts; however, the final viability percentages of this organism did not differ significantly in the probiotic fermented milks tested. The cultured dairy foods made from cow, sheep, and goat milks had comparable B. animalis ssp. lactis BB-12 counts on d 0, exceeding by approximately 0.5 log₁₀ cycle those in the camel milk-based product. No significant losses occurred in viability of bifidobacteria in fermented camel, cow, and sheep milks during 6 wk of refrigerated storage. In conclusion, all 4 varieties of milk proved to be suitable raw materials for the manufacture of ABT-type fermented dairy products that were microbiologically safe and beneficial for human consumption. It was suggested that milk from small ruminants be increasingly used to produce probiotic fermented dairy foods. The development of camel milk-based probiotic cultured milks appears to be even more promising because new markets could thus be conquered. It must be emphasized, however, that further microbiological and sensory studies, technology development activities, and market research are needed before such food products can be successfully commercialized.     

Effect of acacia (Robinia pseudo-acacia L.) honey on the characteristic microflora of yogurt during refrigerated storage

The primary purpose of this research was to monitor the influence of acacia honey addition to yogurt milk on survival of the microbial flora of yogurt during refrigerated storage for 6 wk. Results showed that the presence of honey at 1.0% to 5.0% (w/v) did not significantly influence (P>0.05) the viability of characteristic microorganisms (i.e., Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) in yogurt during storage at 4 degrees C. Similarly, honey had no effect on pH and lactic acid levels of the final products. Despite these findings, enrichment of yogurt with honey is recommended because honey is a natural sweetener that possesses a wide range of beneficial nutritional properties. In addition, at a rate of approximately 3.0% (w/v), it highly improves the sensory quality of the finished product without having a detrimental effect on characteristic lactic acid bacteria.     

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.     

Effect of long‐chain inulin on the texture profile and survival of Lactobacillus paracasei ssp. paracasei in set yoghurts during refrigerated storage

The effects of adding inulin at 20 g/L as a fat replacer and probiotic bacteria on the physicochemical and textural characteristics of yoghurt were studied. The ability of long‐chain inulin to improve the probiotic (Lactobacillus paracasei ssp. paracasei) bacteria viability in yoghurt was investigated. The addition of inulin made the texture (firmness, cohesiveness, adhesiveness and gumminess) of skimmed yoghurt similar to that of whole yoghurt, demonstrating the role of inulin as a fat replacer. However inulin increased syneresis and did not influence the viability of probiotic bacteria in the yoghurts. The addition of probiotic bacteria in yoghurts improved syneresis and increased firmness and gumminess.     

Texture changes in fresh cut celery during refrigerated storage

The effect of refrigerated storage temperature (0 or 10 °C) on variations in texture and lignin content of minimally processed celery was assessed. In addition, the evolution of soluble phenols was studied, as well as changes in other quality parameters, namely weight loss, surface colour and contents of chlorophyll and free sugars. To this end, celery cuts were packaged on PVC film‐covered polystyrene trays. The maximum shear force required to cut the strips transversally followed a similar tendency during storage at both temperatures tested. Increases in lignin content were observed which could be related to corresponding texture increases during the first few days in cold storage. There was no evidence that lignification occurred by a cicatrix‐forming process in the wounds caused during cutting. Copyright © 2003 Society of Chemical Industry     

Antioxidant responses in minimally processed celery during refrigerated storage

This work studies the effect of storage temperature (0, 4 and 10 °C) and time on the antioxidant capacity of cut celery packaged in polystyrene trays sealed with PVC film. Samples were taken at 0, 7, 14, 21 and 28 days of storage to determine total phenols, chlorogenic acid and ascorbic acid. The browning potential and antioxidant capacity of the product were also evaluated. The antioxidant power presented similar behaviour for the three temperatures tested, decreasing after the first 7 days and then increasing up to day 14. Such increase coincided with an elevation of the ascorbic acid content, which was stronger for higher temperatures. As a general conclusion, minimally processed celery retained its initial antioxidant capacity for a period of 21 days at 0 °C, showing the lowest levels of browning potential at this temperature.     

Proteolytic degradation of ewe milk proteins during fermentation of yoghurts and storage

Yoghurts are mostly produced from cow milk and to a very limited extent from ewe milk. The evolution of caseins and whey proteins in ovine milk submitted to different thermal treatments (63 degrees C/30 min; 73 degrees C/15 min; 85 degrees C/10 min or 96 degrees C/5 min) was followed during fermentation of yoghurts and during their storage up to 14 days, using two different sets of starters. One set of starter LAB was a "ropy" culture (YC-191), which is a well-defined mixed strain culture containing Streptococcus thermophilus ST-143 and Lactobacillus delbrueckii subsp. bulgaricus (LB-18 and LB-CH2). The other set of starter bacteria (YC-460) was a standard yoghurt culture("non-ropy") containing mixed strain culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. Contents of free amino groups in produced yoghurts increased gradually during the fermentation, up to a maximal value obtained after 4 h fermentation, then they did not change significantly during storage of yoghurt produced with YC-191 starter. In contrary, a large drop in the amount of free amino groups was observed in the first 24 h of storage in the case of yoghurt made with YC-460 indicating that microorganisms continue still to grow in low temperatures. During fermentation and storage of both yoghurt types, alpha-lactalbumin was hydrolyzed to a slightly bigger extent than beta-lactoglobulin. During fermentation, beta-casein was slightly more degraded than alpha(s)-caseins; however, the opposite was observed during storage up to 14 days. Generally, a more intense heat pretreatment led to a higher degradation of whey proteins and caseins during fermentation and storage. Differences in proteolytic activity between the two starters used (whey proteins more degraded by YC-191; caseins more degraded by YC-460) may lead to improvement in production and formulation of yoghurts differing in their physicochemical and rheological properties.     

Viability of bifidobacteria in commercial dairy products during refrigerated storage

Commercial milk and two brands of yogurt containing bifidobacteria were obtained from retail outlets. All products were evaluated for viability of bifidobacteria and lactic acid bacteria during refrigerated storage at 4 degrees C. Milk was evaluated at 9, 6, and 3 days prior and past its expiration date. The yogurts were evaluated at 3, 2, and 1 week prior and past their expiration. Viability of bifidobacteria and lactic acid bacteria in milk and yogurt remained above 10(6) CFU/ml or g until the expiration date of the respective products. This microbial concentration is the recommended minimum dose to receive the health benefits of these organisms.     

Viability of microencapsulated bifidobacteria in set yogurt during refrigerated storage

Bifidobacteria are probiotic organisms that improve the microbial balance in the human gut. They can be incorporated as live cultures in fermented dairy foods, including yogurt, for transmission to humans. Because bifidobacteria are sensitive to high acidity, their viability in yogurt is limited. The objective of the present study was to investigate the effect of microencapsulation on the viability of bifidobacteria in yogurt during refrigerated storage for 30 d. Live bifidobacterial cells were encapsulated in kappa-carrageenan. Cell enumeration, determination of titratable acidity and pH, quantitation of lactic and acetic acids, and sensory evaluation (consumer test) were carried out on the yogurt samples. Microbiological results showed a decline of 78 and 70.5% in the population of Bifidobacterium longum B6 and B. longum ATCC 15708, respectively, for the treatments containing nonencapsulated cells. No difference in bifidobacterial population was observed in the encapsulated treatments. The acetic acid content in the yogurt with nonencapsulated bifidobacteria was higher than that in the plain yogurt (control) and encapsulated treatments. The increase in lactic acid content during storage was not different among the various treatments for B. longum B6, but was greater for nonencapsulated than encapsulated B. longum 15708 and the control. Consumers judged the nonencapsulated treatment as the most sour, which was likely due to the higher acetic acid content. Consumers preferred the control and nonencapsulated treatments over the encapsulated treatment. Microencapsulation appears to increase the viability of bifidobacteria in yogurt. This technique can be used to transmit bifidobacteria via fermented products provided that sensory characteristics of the product are improved or maintained.