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.     

Effect of the Lactobacillus rhamnosus strain GG and tagatose as a synbiotic combination in a dextran sulfate sodium-induced colitis murine model

Synbiotics, a combination of prebiotics and probiotics, produce synergistic effects to promote gastrointestinal health. Herein, we investigated the synbiotic interaction between the Lactobacillus rhamnosus strain GG (LGG; a probiotic strain) and tagatose (a prebiotic) in a dextran sulfate sodium (DSS)-induced colitis murine model. Initially, body weight, food intake, and clinical features were dramatically decreased after treatment with DSS, and the addition of LGG, tagatose, or both ameliorated these effects. In our pyrosequencing analysis of fecal microbiota, DSS treatment increased the abundance of Proteobacteria and decreased that of Firmicutes. When LGG and tagatose were administered as synbiotics, the gut microbiota composition recovered from the dysbiosis caused by DSS treatment. In particular, the abundance of Bacteroides, Lactobacillus, and Akkermansia was significantly associated with probiotic, prebiotic, and synbiotic treatments. Taken together, our results suggest that LGG and tagatose as synbiotics can alleviate colitis, and synbiotics could be applied as dietary supplements in dairy foods such as yogurt and cheese.     

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.     

Viability of Probiotics in Goat Cheese During Storage and Under Simulated Gastrointestinal Conditions

The objective of this study was to evaluate the suitability of two strains of probiotic bacteria (Bifidobacterium animalis subsp. lactis and Lactobacillus rhamnosus) incorporated individually into Boursin-type goat cheese and their in vitro resistance through the passage of the gastrointestinal tract in the cheese. The viability of B. lactis and Lb. rhamnosus cultures was unaffected throughout 35 days of storage at 4 °C, with a final count of >?7 log CFU/g. No significant difference (p?>?0.05) was observed between probiotic treatments and control in relation to pH and titratable acidity. B. animalis presented greater resistance to the artificial gastric and enteric juices than Lb. rhamnosus, with mean decreases in the initial populations of 0.2 and 4.0 log CFU/g within 35 days of storage, respectively. The addition of probiotic cultures did not affect the consumer acceptance of the goat cheeses. Three segments of consumers with different liking were identified. The results demonstrated that “Boursin” goat cheese is a promising matrix for the incorporation and protection of B. animalis subsp. lactis.     

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.     

Dynamics of starter,adjunct non-starter lactic acid bacteria and propionic acid bacteria in low-fat and full-fat Dutch-type cheese

The microbial dynamics of Dutch-type cheeses differing in starter (commercial DL starter or single strain of Lactococcus lactis ssp. cremoris), adjunct (Lactobacillus or Propionibacterium) and fat contents (10% or 28% fat) were investigated by culture-dependent and culture-independent analysis. The cheese microbiota was dominated by the adjunct Lactobacillus after 4 weeks of ripening and the fat content did not influence the microbial diversity. The Leuconostoc sp., presumably from the DL starter, was detected in cheeses made with added Lactobacillus plantarum and Lactobacillus rhamnosus and was not detected in cheese made with added Lactobacillus paracasei after 4 and 7 weeks. No Lactobacillus spp. were detected in cheese with added Propionibacterium, while Leuconostoc was the only species detected. In cheeses made with Lc. lactis ssp. cremoris as starter, the Lactobacillus microbiota was similar to the cheese milk microbiota after 24 h while after 4 weeks different species of Lactobacillus and Leuconostoc were detected.     

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.     

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.     

Lactobacillus GG—a human probiotic strain with thorough clinical documentation

Abstract

Probiotic bacteria for human nutrition are generally strains of lactic acid bacteria or bifidobacteria. The beneficial effects of these strains on human health and well‐being are documented in experimental and clinical studies. Lactobacillus (casei subsp.) rhamnosus GG (ATCC 53103) is a probiotic strain of human origin. The strain has most of the characteristics generally proposed for a good probiotic strain, including excellent survival in the stomach and small intestine and transient colonization of the gastrointestinal tract, which is based on its adhesion capacity to intestinal cells. Successful recovery of the strain in stool samples has made dose‐response studies possible and enabled the evaluation of effective dosing using food products. The beneficial effects of the strain have been shown in many types of intestinal disturbances caused by pathogenic bacteria and viruses, as well as in prophylactic use. The latest studies on Lactobacillus GG have shown stabilizing effect on gut permeability and suppression of allergic reactions in food hypersensitivity. The safety of Lactobacillus GG has been documented in experimental, clinical, and epidemiological studies. The scientific documentation of Lactobacillus GG is reviewed in reference to uses in functional and clinical foods.     

Short communication: In vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1 isolated from Minas artisanal cheese

Some Lactobacillus strains may contribute to the health of the host when administered in adequate concentrations, demonstrating their probiotic potential. In contrast, Listeria monocytogenes is a foodborne pathogen that can cause enteropathy, meningoencephalitis, abortion, and septicemia. The aim of this survey was to evaluate the in vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1, isolated from Minas artisanal cheese of the Serra da Canastra (Minas Gerais, Brazil), against Lis. monocytogenes. We submitted B7 and D1 to in vitro testing (antibiogram, tolerance to bile salts and artificial gastric fluid, and spot-on-lawn) and in vivo testing (relative weight gain in mice). Both Lactobacillus strains demonstrated in vitro inhibitory activity against Lis. monocytogenes, as well as sensitivity to antimicrobials and resistance to gastric acids and bile salts. In the in vivo assays, mice treated with D1 gained more weight than mice in the other groups. These results indicate that D1 could have higher probiotic potential than B7 because improvements in feed conversion may help animals fight infection.     

Probiotic potential of Lactobacillus paracasei FM-LP-4 isolated from Xinjiang camel milk yoghurt

Fifty three probiotic lactic acid bacteria (LAB) strains with high anti-oxidative activity from Xinjiang camel milk yoghurt were screened and identified. Four strains showed higher 2,2-diphenyl-1-picrylhydrazyl scavenging activity and reducing power than the reference strain Lactobacillus rhamnosus GG (ATCC53103) with strain FM-LP-4, identified as Lactobacillus paracasei by 16S rDNA nucleotide sequencing, showing the highest. Strain FM-LP-4 also showed higher stress tolerance properties (acidity, bile, and osmotic pressure) and adhesion properties than L. rhamnosus GG. The in vivo antioxidant activity of FM-LP-4 was further assayed in d-galactose-induced oxidative damaged mice. Administration of FM-LP-4 significantly enhanced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and inhibited the biosynthesis of malondialdehyde (MDA) and protein carbonyl in a dose-dependent manner. Additionally, FM-LP-4 strain could significantly decrease the endotoxin level in the serum of mice. Overall, strain FM-LP-4 is a promising probiotic candidate for preparations of functional foods and antioxidant supplements.     

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.     

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.     

Technological and probiotic role of adjunct cultures of non-starter lactobacilli in soft cheeses

The influence of two cheese-isolated Lactobacillus strains on cheese composition, acceptability and probiotic capacity was assessed. Soft cheeses with and without the addition of Lactobacillus plantarum I91 or Lactobacillus paracasei I90 were prepared. Gross composition was assessed and secondary proteolysis was described by soluble fractions and free amino acids profiles. Acceptability was determined by a panel of 98 non-trained consumers. Cheeses harboring added Lactobacillus strains were also studied in vivo to evaluate their probiotic capacity. Gross composition of the cheeses was similar for control and treated (Lactobacillus-added) cheeses. Peptidolysis increased in cheeses with added lactobacilli, which was evidenced by a higher free amino acid content. Overall, the acceptability of the cheeses was good: 65%–80% of the consumers said that they “liked very much” or “liked” the cheeses. Cheeses with L. plantarum I91 showed the highest changes in composition and proteolysis and were the most accepted ones. On the contrary, composition of cheeses with L. paracasei I90 was similar to that of the controls, but these samples were less accepted than cheeses without lactobacilli. The oral administration of cheese containing L. plantarum I91 or L. paracasei I90 proved to be safe and able to enhance the number of IgA + cells in the small intestine lamina propria of mice. The use of selected strains of NSLAB exerted a technological and probiotic role: it contributed to the standardization of cheese quality and induced benefic health effects at the gut mucosa in vivo.     

Influence of whey-based fruit juice containing Lactobacillus rhamnosus on intestinal well-being and humoral immune response in healthy adults

The ability of probiotics to balance intestinal microbes and to modulate gut immune system has been reported for several probiotic strains. In the present study, Lactobacillus rhamnosus strain VTT E-97800 (E800) or L. rhamnosus Lc705 (the latter in combination with Propionibacterium freudenreichii ssp. shermanii JS) were separately administered to healthy adult volunteers (n=11 in both study groups) in a whey-based fruit juice. Survival of the potentially probiotic L. rhamnosus strains in the GI-tract and their effect on intestinal well-being and peripheral immune parameters were assessed. Both L. rhamnosus strains were recovered in high numbers in faecal samples during the consumption period and with few exceptions they were not detected after the 2 weeks follow-up period. Consumption of the juice supplemented with either of the L. rhamnosus strains did not have significant effect on faecal consistency or defecation frequency. No changes were observed in the immunological parameters (number or immunoglobulin A-, G- and M-secreting cells, expression of Fcα-receptor or complement receptors 1 and 3), except for a slight decrease in the number of IgA secreting cells during ingestion of the juice supplemented with L. rhamnosus Lc705 and P. freudenreichii ssp. shermanii JS. The results indicate that L. rhamnosus strains E800 and Lc705 had good survival ability in the GI-tract when administered in a whey-based fruit juice matrix. No adverse effects on intestinal function or on studied immunological parameters were observed during consumption of the whey based juice drink supplemented with the potential probiotic strains.     

Persistence of probiotic strains in the gastrointestinal tract when administered as capsules, yoghurt, or cheese

Most clinical studies of probiotics use freeze-dried, powdered bacteria or bacteria packed in capsules. However, probiotics are commercially available in various food matrices, which may affect their persistence in the gastrointestinal tract. The objective of the study was to compare oral and faecal recovery during and after administration of a combination of Lactobacillus rhamnosus GG and LC705, Propionibacterium freudenreichii subsp. shermanii JS, and Bifidobacterium animalis subsp. lactis Bb12 as capsules, yoghurt, or cheese. This randomized, parallel-group, open-label trial (n = 36) included a 4-week run-in, 2-week intervention, and 3-week follow-up period. Participants consumed 10¹⁰ cfu/day of probiotic combination and provided saliva and faecal samples before, during, and after the intervention. Strain-specific real-time PCR was used to quantify the strains.L. rhamnosus GG was the only probiotic strain regularly recovered in saliva samples. During the intervention period it was recovered in the saliva of 88% of the volunteers at least once. No difference was found between the yoghurt and cheese groups. At the end of the intervention, L. rhamnosus GG and LC705 counts were high in faecal samples of all product groups (8.08 and 8.67 log₁₀ genome copies/g, respectively). There was no matrix effect on strain quantity in faeces or the recovery time after ceasing the intervention. For P. freudenreichii subsp. shermanii JS and B. animalis subsp. lactis Bb12, a matrix effect was found at the end of the intervention (P < 0.01 and P < 0.001, respectively) and in the recovery time during follow-up (P < 0.05 for both). Yoghurt yielded the highest faecal quantity of JS and Bb12 strains (8.01 and 9.89 log₁₀ genome copies/g, respectively). The results showed that the administration matrix did not influence the faecal quantity of lactobacilli, but affected faecal counts of propionibacteria and bifidobacteria that were lower when consumed in cheese. Thus, the consumption of probiotics in yoghurt matrix is highly suitable for studying potential health benefits and capsules provide a comparable means of administration when the viability of the strain in the capsule product is confirmed.     

Prato cheese containing Lactobacillus casei 01 fails to prevent dextran sodium sulphate-induced colitis

The role of experimental probiotic Prato cheese containing Lactobacillus casei 01 in the prevention of dextran sodium sulphate (DSS)-induced ulcerative colitis in mice was evaluated. For the DSS in vivo model, mice were divided into six groups. Groups 1–3 represented noninflamed groups and groups 4–6 received a DSS (2%) solution. Mice from groups 1 and 4 intragastrically received 500 μL of phosphate-buffered saline (control groups), while mice from groups 2 and 5 intragastrically received 500 μL of conventional Prato cheese (control groups). Finally, mice from group 3 and 6 intragastrically received 500 μL of Prato probiotic cheese containing L. casei 01 (9.47 log cfu mL⁻¹). Groups treated with probiotic Prato cheese exhibited reduced weight loss caused by the consumption of DSS solution. However, the probiotic cheese failed to reduce the inflammation scores in DSS-induced colitis group. Overall, probiotic Prato cheese was not effective to ameliorate the symptoms of DSS-induced colitis.     

Effect of clinical and probiotic Lactobacillus rhamnosus strains on intestinal permeability and bacterial translocation in healthy and colitic rats

Lactobacillus rhamnosus is one of the most widely used probiotic microorganisms. Although this microorganism is not known for its virulence some rare cases of bacteraemia and endocarditis have been observed. Thus, it is important to identify the possible risks associated with each strain. Comparison of clinical and probiotic strains may give information on properties that could be a safety concern. A possible adverse effect is bacterial translocation. Animals with induced-colitis constitute a good model to assess translocation in the case of mucosal barrier disruption. We aimed at determining the ability of five L. rhamnosus strains, from clinical or probiotic origin, showing different in vitro properties, to induce in vivo translocation in both, healthy and colitic animals. We also tested the effects on the gut mucosal lining by measuring intestinal permeability. None of the in vitro parameters used for selection of the strains included in this study appear to be a risk for translocation or mucosal barrier disruption.     

Probiotics: An alternative strategy for combating salmonellosis: Immune mechanisms involved

Salmonella produces infections of different nature and severity depending of many factors including the Salmonella serovar involved, strain virulence, infective dose, host animal species, age and immune status of the host. The treatments against Salmonella infections rely on supportive and antibiotic therapy to eliminate the pathogen, but the development of resistance by Salmonella to the antimicrobials most commonly used limits its efficacy. Other disadvantages of antibiotic treatments are that they can lead to acute diarrhea (antimicrobials normally induce an imbalance of intestinal bacterial flora) and may produce chronic toxicity. Considering this undesired consequences of antibiotics and because at the present there are no effective oral vaccines which protect against salmonellosis, scientists have been searching for alternative methods to control enteric infections. In the present review, probiotics are proposed as an attractive possibility to attend this concern. Probiotic are live microorganisms, which when administered in adequate amounts confer a health benefit on the host. In vitro and in vivo studies showed the effectiveness of probiotic administration in the prevention or in the treatment against Salmonella infection. There are several mechanisms by which probiotic strains might exert their effects. They include non immune mechanisms (stabilization of the gut mucosal barrier, competition for adhesion, secretion of antimicrobial substances, etc.) and the modulation of the mucosal and systemic immune responses. These mechanisms are species and/or strain specific. There are also evidences that in some cases, a mix of probiotic strains can be more useful than each strain alone against this infection. In addition, the presence of one or more probiotic strains in a fermented product can improve the beneficial properties of the probiotic strains involved. It was also reviewed the security of probiotics administration after Salmonella infection in healthy host and in immunosuppressed or babies hosts. Although, the major part of the researches were performed in animal models through in vivo assays or by in vitro studies using human cell lines, some studies carried out in humans to verify the probiotic effects were also addressed in the present review. Nevertheless, is of critical importance to perform more clinical trials in humans to validate the results obtained with each specific probiotic strain or probiotic product.