Viability of the Lactobacillus rhamnosus HN001 Probiotic Strain in Swiss‐ and Dutch‐Type Cheese and Cheese‐Like Products

The objective of this study was to determine the viability of the probiotic Lactobacillus rhamnosus HN001 in Swiss‐type and Dutch‐type cheese and cheese‐like products (milk fat is substituted by stearin fraction of palm fat) during manufacture, ripening, and storage. The use of the probiotic L. rhamnosus HN001 in Dutch‐type cheese and cheese‐like products significantly (P = 0.1) changed their chemical composition (protein and fat content) and an insignificant increase (approximately 1.6% in cheese‐like products and approximately 0.3% in cheese) in yield. L. rhamnosus HN001 did not affect the rate of changes in the pH of ripened cheese and cheese‐like products. A minor increase in probiotic counts was observed in initial stages of production and were partially removed with whey. Ripened cheese and cheese‐like products were characterized by high survival rates of probiotic bacteria which exceeded 8 log CFU/g after ripening. An insignificant reduction in the number of viable probiotic cells was noted during storage of Swiss‐type and Dutch‐type cheese, whereas a significant increase in probiotic cell counts was observed in cheese‐like products during storage.     

Assessment of the probiotic potential of lactic acid bacteria isolated from Minas artisanal cheese produced in the Campo das Vertentes region,Brazil

The microbiota of the Brazilian Minas artisanal cheese, made from raw milk, is not well known and may include probiotic bacteria. This study aimed to assess the in vitro and in vivo probiotic properties of the lactic acid bacteria isolated from these cheeses. Thirty‐six samples of the Lactobacillus/Pediococcus group were selected for in vitro investigation. Pediococcus acidilactici (PA2) and Lactobacillus plantarum (LP4) showed the best results and were tested for their ability to protect Salmonella Typhimurium orally infected mice. LP4 showed better probiotic potential than PA2 and allowed higher values of weight gain (P < 0.05). Thus, Lb. plantarumLP4 may have potential use as a probiotic in foods after future technological screening.     

Survival and activity of 5 probiotic lactobacilli strains in 2 types of flavored fermented milk

The viability of 5 probiotic lactobacilli strains (Lactobacillus acidophilus LA-5, Lactobacillus casei L01, Lactobacillus casei LAFTI L26, Lactobacillus paracasei Lcp37, and Lactobacillus rhamnosus HN001) was assessed in 2 types of probiotic flavored drink based on fermented milk during 21 days of refrigerated storage (5°C). Also, changes in biochemical parameters (pH, titrable acidity, and redox potential) during fermentation as well as the sensory attributes of final product were determined. Among the probiotic strains, L. casei LAFTI L26 exhibited the highest retention of viability during refrigerated storage period, while L. acidophilus LA-5 showed the highest loss of viability during this period. The decline in cell count of probiotic bacteria in strawberry fermented milk was significantly greater compared to peach fermented milk. In an overall approach, peach fermented milk containing L. casei LAFTI L26 was selected as the optimal treatment in this study in both aspects of viability and sensory accpeptibility.     

Survival of Lactobacillus rhamnosus strains inoculated in cheese matrix during simulated human digestion

Survival of probiotic bacteria during transit through the gastrointestinal (GI) tract is influenced by a number of environmental variables including stomach acidity, bile salts, digestive enzymes and food matrix. This study assessed survival of seven selected Lactobacillus rhamnosus strains delivered within a model cheese system to the human upper GI tract using a dynamic gastric model (DGM). Good survival rates for all tested strains were recorded during both simulated gastric and duodenal digestion. Strains H12, H25 and N24 demonstrated higher survival capacities during gastric digestion than L. rhamnosus GG strain used as control, with H12 and N24 continuing to grow during duodenal digestion. Strains L. rhamnosus F17, N24 and R61 showed adhesion properties to both HT-29 and Caco-2 cells. The ability to attach to the cheese matrix during digestion was confirmed by scanning electron microscopy, also indicating production of extracellular polysaccharides as a response to acid stress.     

Prato cheese as suitable carrier for Lactobacillus acidophilus La5 and Bifidobacterium Bb12

To assess Prato cheese as suitable carrier for probiotic bacteria, four cheeses were produced. Control cheese contained only starter culture, whereas the others contained starter culture and the probiotic cultures Lactobacillus acidophilus La5 and Bifidobacterium Bb12 either separately or in combination. Bacterial viability, physicochemical composition, proteolysis, and texture profile were assessed over 60 days of storage. The addition of microorganisms together or separately did not affect the characteristics of Prato cheese. On storage, the cheeses showed increased proteolysis, lower firmness, and the probiotic cheeses presented counts higher than 10⁶ cfu g⁻¹. The viability of probiotics during in vitro gastrointestinal simulation, including the effect of the cheese matrix, was also assessed. The probiotic bacteria showed resistance to loss of viability during in vitro gastrointestinal simulation.     

Isomaltooligosaccharide increases the Lactobacillus rhamnosus viable count in Cheddar cheese

Five batches of Cheddar cheese were manufactured containing different levels of isomaltooligosaccharide (IMO) and a probiotic strain of Lactobacillus rhamnosus to study the effect of IMO on the survival of starter lactococci and probiotic micro‐organisms, on proteolytic patterns, cheese composition and sensory properties. The cheese was exposed to conditions simulating those found in the gastrointestinal tract to evaluate the survival of Lb. rhamnosus. Results demonstrated that the addition of Lb. rhamnosus and IMO did not affect the main compositional variables of Cheddar cheese. The counts of starter culture and probiotic organisms increased in cheese which contained Isomaltooligosaccharide (Batches 3, 4 and 5) more than in the control (Batches 1 and 2) during the fermentation. The probiotic counts in fresh cheese (B‐4) was 9.23 log₁₀ cfu/g which was more than one log cycle greater than in the control (B‐2). The probiotic counts remained above 8 log₁₀ cfu/g at the end of the manufacturing process. Primary proteolysis was not affected by the addition of probiotic bacteria and IMO, but the level of secondary proteolysis was slightly higher compared with the control group. The addition of IMO improved the texture and sensory quality of the cheese and the probiotic bacterium had the same effect. Under conditions that simulated the gastrointestinal tract, the probiotic bacteria in cheese (B‐4) exhibited good survival and remained above the recommended 6–7 log₁₀ cfu/g.     

A survey of non-starter lactic acid bacteria in traditional cheeses: Culture dependent identification and survival to simulated gastrointestinal transit

Cultivable NSLAB in traditional Pasta filata and ewes' milk cheeses were studied by both PCR-DGGE of cells from Rogosa agar and by isolation and molecular identification after a simulated gastric juice (SGJ) treatment of the cheese. Two to six species were retrieved from each sample. The majority of isolates were identified as Lactobacillus paracasei, Lactobacillus fermentum, Lactobacillus rhamnosus, Enterococcus or Pediococcus. Bile tolerance and bile salt hydrolase (BSH) activity were tested on 88 strains: 64% were able to grow with ≥0.15% bile and 40% were BSH positive. The effect of simulated digestion was tested on 15 strains. Inactivation ranged from 0.15 to 2.93 log cycles; most of the lethality was associated with pancreatic juice treatment. Although SGJ treatment alone may not provide a correct estimate of tolerance to gastrointestinal transit, it allowed selection of strains with a high tolerance to gastric juice, which may be tested as probiotic candidates.     

Development of a fermented goats' milk containing Lactobacillus rhamnosus: in vivo study of health benefits

BACKGROUND: Lactobacillus rhamnosus CRL1505, a strain of goats' milk origin, is able to stimulate mucosal immunity and protect immunocompetent mice from intestinal and respiratory infections. RESULTS: In this work we developed and characterized a fermented goats' milk containing L. rhamnosus CRL1505, and we demonstrated in a model of immunosuppression in mice that the final dairy product preserves the immunomodulatory properties of the strain. L. rhamnosus CRL1505 survived the manufacturing process of fermented milk and maintained a viability of 10⁶ cfu g^?1 during storage. The fermented goats' milk was accepted by 90.48% of the panelists and was considered as having an acid taste and pleasant aroma. We also demonstrated that the developed product, used as a supplement during the repletion of immunocompromised malnourished mice, was effective in accelerating the recovery of clinical parameters altered by malnutrition and to induce increased resistance against intestinal and respiratory infections. CONCLUSION: Goats' milk fermented with L. rhamnosus CRL1505 could be manufactured as an alternative probiotic dairy product since this new food has the ability to stimulate the common mucosal immune system and to improve defenses against respiratory and intestinal infections. Copyright © 2011 Society of Chemical Industry     

A comprehensive approach to determine the probiotic potential of human-derived Lactobacillus for industrial use

Specific strains should only be regarded as probiotics if they fulfill certain safety, technological and functional criteria. The aim of this work was to study, from a comprehensive point of view (in vitro and in vivo tests), three Lactobacillus strains (Lactobacillus paracasei JP1, Lactobacillus rhamnosus 64 and Lactobacillus gasseri 37) isolated from feces of local newborns, determining some parameters of technological, biological and functional relevance. All strains were able to adequately grow in different economic culture media (cheese whey, buttermilk and milk), which were also suitable as cryoprotectants. As selective media, LP-MRS was more effective than B-MRS for the enumeration of all strains. The strains were resistant to different technological (frozen storage, high salt content) and biological (simulated gastrointestinal digestion after refrigerated storage in acidified milk, bile exposure) challenges. L. rhamnosus 64 and L. gasseri 37, in particular, were sensible to chloramphenicol, erythromycin, streptomycin, tetracycline and vancomycin, increased the phagocytic activity of peritoneal macrophage and induced the proliferation of IgA producing cells in small intestine when administered to mice. Even when clinical trails are still needed, both strains fulfilled the main criteria proposed by FAO/WHO to consider them as potential probiotics for the formulation of new foods.     

Probiotic lactobacilli in a semi-soft cheese survive in the simulated human gastrointestinal tract

In this study, probiotics Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus HN001 in cheese were studied using models simulating the human gastrointestinal tract with the aim of investigating whether the cheese matrix affected the survival and metabolic properties of these probiotic strains. Probiotics in cheese survived in the simulated upper gastrointestinal tract model, and numbers of L. acidophilus, L. rhamnosus and total lactobacilli were increased in the colonic fermentation simulations of the probiotic cheese when compared with the non-probiotic cheese used as a control. The cheese matrix also beneficially affected cyclooxygenase-gene expression of colonocytes in a cell culture model. Freeze-dried probiotics, which were also analysed in the colonic simulator, showed similar changes in Lactobacillus numbers, although gave a stronger increase and also affected other microbial groups. These results indicate that the probiotic microbes in cheese survive in the gastrointestinal tract and that the cheese matrix does not seem to affect the probiotic survival.     

Effects of tropical fruit pulps and partially hydrolysed galactomannan from Caesalpinia pulcherrima seeds on the dietary fibre content,probiotic viability,texture and sensory features of goat dairy beverages

The use of cheese whey and probiotic cultures in the production of dairy beverages has been highly attractive; nonetheless, whey-based goat beverages tend to be poor and watery when compared to fermented milks. The addition of fruits and fibre ingredients might improve texture and mouthfeel of this kind of product. Fermented whey-based goat beverages prepared using Streptococcus thermophilus TA-40 as starter culture, with added guava or soursop pulps, and with or without addition of partially hydrolysed galactomannan from Caesalpinia pulcherrima seeds (PHGM), showed to be good vehicles for Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus rhamnosus Lr-32, maintaining their viability above 7 log CFU/ml during 21 days. PHGM increased the dietary fibre content and enhanced the instrumental texture and sensory features of both guava and soursop dairy beverages, especially texture, appearance, and overall acceptability. The PHGM might be recommended to improve nutritional and sensory quality of fermented probiotic beverages produced with goat milk and cheese whey.     

In vitro evaluation of physiological probiotic properties of different lactic acid bacteria strains of dairy and human origin

Eleven lactic acid bacteria strains of importance to the dairy industry were subjected to in vitro analyses to determine their probiotic potential. Seven strains were isolated from ewe’s and cow’s milk (Enterococcus faecalis – five –, Lactococcus lactis and Lactobacillus paracasei). Four were obtained from American Type Culture Collection (ATCC), isolated from cheese (Lactobacillus casei 393), human feces (L. paracasei 27092 and Lactobacillus rhamnosus 53103) and used in cheese making (L. lactis 54104). Although none of the strains was able to degrade mucin, all E. faecalis showed, at least, one transferable antibiotic resistance, which excluded them as candidates for addition to foods. Of the remaining six safe strains, L. lactis strains were more tolerant to low pH than Lactobacillus spp.; all were tolerant to pancreatin and bile salts and showed antibacterial activity. The highest level of adhesion to Caco-2 cells was observed with L. lactis 660, even higher than L. rhamnosus ATCC 53103 (recognized probiotic and used as control). The physiological probiotic properties of these strains, mainly isolated from dairy sources, are interesting in view of their use in cheese productions as starter and non starter cultures. The five LAB safe strains studied may have potential as novel probiotics in the dairy foods.     

Flavour-enhancement of low-fat Kashkaval cheese using heat-or freeze-shocked Lactobacillus delbrueckii var. helveticus cultures

Kashkaval cheese of different fat contents was manufactured using heat- or freeze-shocked cultures of Lactobacillus delbrueckii var. helveticus added at a level of 2% to cheese milk prior to renneting. Levels of moisture, total N, salt or titratable acidity of cheeses with different fat levels were different. Proteolysis and lipolysis in low-fat Kashkaval cheese without additives were lower than those found in full-fat cheese. Incorporation of heat- or freeze-shocked L. delbrueckii var. helveticus cultures into milk of low-fat cheese greatly enhanced proteolysis and increased slightly the levels of free fatty acids. Regular low fat cheese did not develop adequate Kashkaval cheese flavour and the cheese was firm. However, addition of heat- or freeze-shocked cultures increased the flavour intensity and improved body and texture of the resultant cheese, yielding low-fat cheese with flavour intensity and body characteristics similar to those of standard fat cheese at each stage of ripening.     

Hazelnut milk fermentation using probiotic Lactobacillus rhamnosus GG and inulin

Following the consumer demand of healthy vegetable products due to their interesting nutritional profiles and potential functionalities, the fermentation process of hazelnut milk with Lactobacillus rhamnosus GG and S. thermophilus was studied. The effect of different factors (glucose, inulin and inoculum contents) was analysed to ensure sufficient probiotic survivals in a minimum time. The shelf life of the optimised product was characterised in terms of its main physicochemical and quality parameters (probiotic survivals and sensory analysis). Results showed that the defined formulation allowed high probiotic survivals (≈10⁸ cfu mL^?1) throughout cold storage and >60% survived to the in vitro digestion process (≈10⁵ cfu mL^?1). Lactobacillus rhamnosus GG was no able to degrade inulin, which remained to exert health benefits in the host. The product was highly appreciated by the sensory panel during its shelf life despite the formation of a weak gel, which presented syneresis at the last storage time.     

Effect of fermented milk containing Lactobacillus rhamnosus SD11 on oral microbiota of healthy volunteers: A randomized clinical trial

The aims of this study were to evaluate whether short-term consumption of fermented milk containing Lactobacillus rhamnosus SD11 affected levels of oral microbiota in vivo and whether L. rhamnosus SD11 could colonize in the human mouth. We also monitored for potential side effects of the probiotic. The applicability of using L. rhamnosus SD11 compared with Lactobacillus bulgaricus as a starter culture for fermented milk was evaluated. After informed consent, 43 healthy young adults were recruited and randomly assigned to either the probiotic or control group and received fermented milk containing L. rhamnosus SD11 or L. bulgaricus, respectively, once daily for 4 wk. The numbers of mutans streptococci, lactobacilli, and total bacteria in saliva were counted at baseline and then after 4 and 8 wk. An oral examination was performed at baseline and after 8 wk. The persistence of L. rhamnosus SD11 was investigated by DNA fingerprinting using arbitrary primer-PCR. Results demonstrated that statistically significant reductions in mutans streptococci and total bacteria were observed in the probiotic group compared with the control group, and the number of lactobacilli was significantly increased in both groups after receiving fermented milks. Lactobacillus rhamnosus SD11 could be detected (in >80% of subjects) up to 4 wk following cessation of dosing among subjects in the probiotic group. No side effects were reported. Thus, L. rhamnosus SD11 could be used as a starter culture for fermented milk. Daily consumption of L. rhamnosus SD11-containing fermented milk for 4 wk may have beneficial effects on oral health by reducing salivary levels of mutans streptococci. The probiotic was apparently able to colonize the oral cavity for a longer time than previously reported. However, the potential benefits of probiotic L. rhamnosus SD11 on oral health require further evaluation with a larger group of volunteers in a longer-term study.     

Survival of probiotic lactobacilli in the upper gastrointestinal tract using an in vitro gastric model of digestion

The aim of this study was to investigate survival of three commercial probiotic strains (Lactobacillus casei subsp. shirota, L. casei subsp. immunitas, Lactobacillus acidophilus subsp. johnsonii) in the human upper gastrointestinal (GI) tract using a dynamic gastric model (DGM) of digestion followed by incubation under duodenal conditions. Water and milk were used as food matrices and survival was evaluated in both logarithmic and stationary phase. The % of recovery in logarithmic phase ranged from 1.0% to 43.8% in water for all tested strains, and from 80.5% to 197% in milk. Higher survival was observed in stationary phase for all strains. L. acidophilus subsp. johnsonii showed the highest survival rate in both water (93.9%) and milk (202.4%). Lactic acid production was higher in stationary phase, L. casei subsp. shirota producing the highest concentration (98.2 mM) after in vitro gastric plus duodenal digestion.     

Microbial,physico‐chemical and sensory characteristics of mango juice‐enriched probiotic dairy drinks

This study aimed to determine whether mango juice can improve the viability of probiotics in a fermented dairy‐based beverage whilst maintaining its quality characteristics. Formulations containing Lactobacillus acidophilus La‐5 culture, whole cow's milk and varying concentrations of mango juice (0%, 10%, 20%, 30% and 40% (w/w)) were produced and stored for five weeks at 4 °C. Results showed that probiotic viability was enhanced with the addition of 10% mango juice. Additionally, this formulation improved probiotics tolerance when exposed to in vitro gastrointestinal digestion. According to the sensory analysis, beverage sensory scores improved as levels of mango juice increased from 20% to 40%.     

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.     

Effect of different matrices on probiotic resistance to in vitro simulated gastrointestinal conditions

In this study, we evaluated the effect of different matrices (MRS, milk and milk with inulin) on the tolerance of probiotic strains (Lactobacillus acidophilus La‐5 and Bifidobacterium animalis subsp. lactis BB‐12) to simulated conditions similar to those found in the gastrointestinal tract. Both probiotic strains demonstrated a significantly lower viability after exposure to in vitro gastric and intestinal conditions, and B. animalis subsp. lactis BB‐12 showed higher survival during the test compared to L. acidophilus La‐5 in all tested matrices. Milk and inulin protected probiotics from in vitro gastrointestinal stress. These results suggest that it is critical to formulate the food matrix to be used as probiotic carrier.     

Effect of buckwheat flour and oat bran on growth and cell viability of the probiotic strains Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their combination SYNBIO®, in synbiotic fermented milk

Fermented foods have a great significance since they provide and preserve large quantities of nutritious foods in a wide diversity of flavors, aromas and texture, which enrich the human diet. Originally fermented milks were developed as a means of preserving nutrients and are the most representatives of the category. The first aim of this study was to screen the effect of buckwheat flour and oat bran as prebiotics on the production of probiotic fiber-enriched fermented milks, by investigating the kinetics of acidification of buckwheat flour- and oat bran-supplemented milk fermented by Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their 1:1 combination named SYNBIO®. The probiotic strains viability, pH and sensory characteristics of the fermented fiber-enriched milk products, stored at 4 °C for 28 days were also monitored. The results showed that supplementation of whole milk with the tested probiotic strains and the two vegetable substrates results in a significant faster lowering of the pH. Also, the stability of L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® during storage at 4 °C for 28 days in buckwheat flour- and oat bran-supplemented samples was remarkably enhanced. The second aim of the study was to develop a new synbiotic product using the best combination of probiotics and prebiotics by promoting better growth and survival and be acceptable to the consumers with high concentration of probiotic strain. This new product was used to conduct a human feeding trial to validate the fermented milk as a carrier for transporting bacterial cells into the human gastrointestinal tract. The probiotic strains were recovered from fecal samples in 40 out of 40 volunteers fed for 4 weeks one portion per day of synbiotic fermented milk carrying about 10⁹ viable cells.