Comparative evaluation of yogurt and low-fat cheddar cheese as delivery media for probiotic Lactobacillus casei

This study used Lactobacillus casei 334e, an erythromycin-resistant derivative of ATCC 334, as a model to evaluate viability and acid resistance of probiotic L. casei in low-fat Cheddar cheese and yogurt. Cheese and yogurt were made by standard methods and the probiotic L. casei adjunct was added at approximately 10(7) CFU/g with the starter cultures. Low-fat cheese and yogurt samples were stored at 8 and 2 degrees C, respectively, and numbers of the L. casei adjunct were periodically determined by plating on MRS agar that contained 5 microg/mL of erythromycin. L. casei 334e counts in cheese and yogurt remained at 10(7) CFU/g over 3 mo and 3 wk, respectively, indicating good survival in both products. Acid challenge studies in 8.7 mM phosphoric acid (pH 2) at 37 degrees C showed numbers of L. casei 334e in yogurt dropped from 10(7) CFU/g to less than 10(1) CFU/g after 30 min, while counts in cheese samples dropped from 10(7) CFU/g to about 10(5) after 30 min, and remained near 10(4) CFU/g after 120 min. As a whole, these data showed that low-fat Cheddar cheese is a viable delivery food for probiotic L. casei because it allowed for good survival during storage and helped protect cells against the very low pH that will be encountered during stomach transit.     

Evaluation of alginate-pectin capsules in Cheddar cheese as a food carrier for the delivery of folic acid

Edible gums were evaluated for folic acid encapsulation efficiency as single and mixed polymers. Alginate and pectin yielded the highest encapsulation efficiencies, 216 and 196 μg/100 g, respectively. Upon being combined and the encapsulation conditions optimized, the alginate-pectin (alg-pect) mixture resulted in 360 μg/100 g of folic acid being encapsulated. The alg-pect capsules were tested for their stability in a milk system where pH was adjusted from 6.7 to 4.5 over a 4 h period. Folic acid retention under the test conditions was retained at 360 μg/100 g within the alg-pect capsules indicating their ability to remain intact in a milk system. Stress tolerance of the capsules was tested under simulated cheese press pressures for 4 h until a force of 0.814 N/cm² was detected. Folic acid retention of up to 288 μg/100 g was recorded for the alg-pect capsules. Three stages in Cheddar cheese manufacturing namely; addition of capsules in the milk, incorporation of capsules after milling the curd and injection into the pressed block of raw cheese, were studied for capsules distribution. The former had the most even distribution of capsules while the latter two had poor and irregular distribution. Encapsulated folic acid showed more stability (360 μg/100 g) than the free folic acid (109 μg/100 g) in Cheddar cheese during the 3 months ripening period.     

Ice-cream as a probiotic food carrier

Ice-creams are food products showing potential for use as probiotic vehicles, with the added advantage of being appreciated by people belonging to all age groups and social levels. However, the development of ice-creams containing probiotic bacteria requires the overcoming of certain technological intrinsic requirements related to their processing stages. The aim of the present paper was to review the technological parameters involved in the production of probiotic ice-creams. Although the application of probiotics in cheeses, and especially in fermented milks, has been widely explored in the literature, ice-cream is a relatively innovative matrix for the application of probiotics, and thus a review about its potential as probiotic food carrier could be very helpful.     

Evaluation of Bifidobacterium breve strain Yakult-fermented soymilk as a probiotic food

We evaluated the effects of Bifidobacterium breve-fermented soymilk on probiotic function. An administered strain of B. breve strain Yakult was capable of growing in soymilk with no additives as high as 10(9) CFU/ml. During storage of the fermented soymilk at 10 degrees C for 20 days, viable counts of the strain did not change. The growth inhibition of the strain in a bile-containing medium was lessened by the addition of soy protein. In human feeding experiments, the administered B. breve was recovered at a level of over 10(9) CFU/g faeces, accompanied by an increase in the total number of bifidobacteria. These results indicate that fermented soymilk with B. breve strain Yakult could be a novel type of probiotic food.     

Survival of probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing in the gastrointestinal tract

Various foods may be used to deliver probiotic bacteria into the gastrointestinal tract; one such example is Requeijão, a Portuguese whey cheese. Survival and stability of Bifidobacterium animalis strains BLC-1, Bb-12, and Bo, Lactobacillus acidophilus strains LAC-1 and Ki, L. paracasei ssp. paracasei strain LCS-1 and L. brevis strain LMG 6906 inoculated into Requeijão, when exposed to simulated gastrointestinal tract conditions, were assessed. Homogenates of inoculated whey cheese in 0.85% (w/v) sterile saline water were exposed to a solution of hydrochloric acid (pH 2.5–3.0) and pepsin (1000 units mL^–1) at 37 °C, and then to 0.3% (w/v) bile salts after 60 or 120 min of acid exposure. All bacterial strains retained their initial viable cell numbers. Bifidobacterium animalis strains Bb-12 and Bo, and L. brevis strain LMG 6906 exhibited the highest viable cell numbers when exposed to bile salts, whereas the other strains had variable death rates.     

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.     

Determination of quantitative food consumption levels for use in microbial risk assessments: cheddar cheese as an example

Microbial risk assessment (MRA) is becoming increasingly used in the management of food safety because it can be used to quantify risks and help rank intervention strategies. The exposure assessment components of the assessments have become complex with many aspects of the contamination, survival, and growth of a pathogen in a food being taken into consideration. Insufficient consumption data constitutes an important data gap and consequently one of many sources of uncertainty in MRA even though the effects of uncertainty are smaller than those affecting bacterial concentration in foods. Therefore, food consumption data also play an important role in exposure assessment of MRA. In the United States, there are large-scale, nationwide sets of consumption data available for use in MRA, i.e., the National Health and Nutrition Examination Survey (NHANES). Newly released dietary interview data in the NHANES 2001 to 2002 survey show that it has been redesigned and that the data were sufficiently updated from previous versions to have more value for MRAs. We propose a model that can effectively use the new data sets and be incorporated into MRAs, using as an example consumption of Cheddar cheese/American-type cheese. This model included the prevalence of food eaten as well as the amount and frequency. We determined the amount of Cheddar/American cheese consumed per day with probability distribution (e.g., lognormal distribution). These could be further determined by gender, age, pregnancy, and combination food type, which we plan to do in the future. The frequency of the range of serving numbers for Cheddar/American cheese consumed per person per day and prevalence as the proportion of a population (e.g., survey respondents) eating a certain food in a day are also presented. Unlike traditional published mean values, the results of this model provide probability distribution intakes that can be compared with mean and median intakes. This allows values in the upper percentiles to be considered for inclusion in MRAs. We believe this simulation model can be adapted with different variables applicable to different foods, pathogens, and specific health risk population groups.     

A washed-curd goat's cheese as a vehicle for delivery of a potential probiotic bacterium: Lactobacillus delbrueckii subsp. lactis UO 004

This study characterizes the probiotic properties of Lactobacillus delbrueckii subsp. lactis UO 004 and examines its suitability for making cheese. This strain was isolated from infant feces and shows interesting features, such as acid and bile tolerance, adherence to intestinal epithelial cells, and inhibition of the growth of certain enteropathogens, that support its potential use as a probiotic strain. In this regard, the suitability of a washed-curd cheese (Vidiago type) made with goat's milk as a delivery system for this probiotic strain was assessed. Lactobacillus delbrueckii subsp. lactis UO 004 was incorporated into a starter culture (IPLA 001). Changes in the overall composition of control and experimental cheeses were determined during ripening through bacteriological, chemical, high-performance liquid chromatography, and gas chromatography analyses. Slight changes in the gross composition and appreciable differences in the flavor compounds profile were observed between control and experimental cheeses. This strain was capable of surviving at high cell numbers (10(8) to 10(9) CFU/g) in cheeses after 28 days of ripening without adversely affecting sensory criteria or appearance of the cheese, thus satisfying the criteria for a probiotic food product.     

Evaluation of a chitosan-based edible film as carrier of natamycin to improve the storability of Saloio cheese

The purpose of this study was to evaluate the effects of the application of chitosan coating containing natamycin on the physicochemical and microbial properties of semi-hard cheese. Three cheese groups were prepared: samples without coating, samples coated with chitosan and with chitosan containing 0.50 mg mL⁻¹ of natamycin, whose minimum inhibitory concentration was previously determinated on cheese surface. Microbiological analyses showed that natamycin coated samples presented a decrease on moulds/yeasts of 1.1 log (CFU g⁻¹) compared to control after 27 days of storage. Addition of natamycin also affected O₂ and CO₂ permeability, increasing from 7.12 to 7.68 × 10⁻¹⁵ g·(Pa s m)⁻¹, and from 10.69 to 64.58 × 10⁻¹⁴ g·(Pa s m)⁻¹, respectively. The diffusion coefficient values of natamycin from the film to phosphate buffered saline solution and to the cheese were 3.60 × 10⁻¹⁰ and 1.29 × 10⁻¹² cm² s⁻¹, respectively. This study demonstrated that chitosan-based coating/films can be used as release system containing natamycin to create an additional hurdle for moulds/yeasts in cheese thus contributing to extend its shelf-life.     

Preparation and characterisation of octenyl succinate starch as a delivery carrier for bioactive food components

The octenyl succinic anhydride (OSA) modified corn starch (OSAS) was synthesised with different degrees of substitution (DS) and the influences of three factors namely OSA/starch ratio, reaction temperature, and reaction time on the DS of OSAS were studied by response surface methodology (RSM). As a delivery carrier material, the characteristics of OSAS were investigated. The resistant starch (RS) content of OSAS was increased with increasing DS, indicating the improvement of colon-targeting property. The swelling ratio (SR) of the OSAS in simulated gastrointestinal fluids was higher than that of native starch and its hydrophilicity was decreased with increasing DS. For in vitro release study, the percentage of released bioactive components was less than 7% within the first 8 h and close to 100% over a period of 36 h when the DS was 0.60. These results demonstrate that OSAS can be a potential carrier for colon-targeted delivery of bioactive food components.     

Use of electron beam irradiation for mold decontamination on cheddar cheese.

Cheddar cheese slices, surface-inoculated with either Penicillium cyclopium or Aspergillus ochraceus spores, were vacuum packaged and irradiated using an electron beam accelerator. Following treatment at .21 and .52 kGy, the shelf-life of cheese containing P. cyclopium was extended by 3 and 5.5 d, respectively, in comparison with inoculated, untreated samples. Under similar treatment and storage conditions, cheese containing A. ochraceus exhibited average shelf-life extensions of 42.5 and 52.2 d, respectively. Increasing the postirradiation storage temperature to 15 degrees C reduced the shelf-life of cheese, especially with samples containing A. ochraceus. The lowest dose required to inactivate ca. 50 to 60 spores/cm2 of either A. ochraceus or P. cyclopium on the surface of cheese was ca. .42 and .95 kGy, respectively. Irradiation survival curves of A. ochraceus and P. cyclopium spores in cheese yielded average values (the dose required to reduce initial population by 90%) of .21 and .42 kGy, respectively.     

Protective effect of whey cheese matrix on probiotic strains exposed to simulated gastrointestinal conditions

A probiotic whey cheese added with Lactobacillus casei LAFTI®L26, Lactobacillus acidophilus LAFTI®L10 or Bifidobacterium animalis Bo was subject in vitro to sequential conditions that parallel the four major steps of digestion: mouth (artificial saliva), oesophagus-stomach (artificial gastric juice), duodenum (artificial intestinal juice) and ileum; its manufacture followed the traditional cheesemaking protocol of Portuguese Requeijão. MRS broth was inoculated in parallel as reference medium, to ascertain the protective effect of the whey cheese matrix itself upon those strains in every digestion step. Mouth conditions had an almost negligible effect upon all three strains, whereas oesophagus-stomach, duodenum and ileum conditions decreased the viable numbers of L. casei and L. acidophilus; in both systems, B. animalis suffered only slight decreases in viable numbers; and L. casei and L. acidophilus behaved likewise in MRS exposed to duodenum and ileum conditions. Whey cheese matrices thus appeared to protect the aforementioned three strains during transit throughout the simulated gastrointestinal system, so they are promising carriers of those probiotic bacteria.     

Evaluation of in vitro probiotic potential of phytase‐producing bacterial strain as a new probiotic candidate

Sixty‐three phytase‐producing bacterial strains were isolated from natural sources, and their probiotic potential was evaluated. Of these, only fifteen strains were selected on the basis of confirmatory plate assay. Among these, five phytase‐producing strains exhibiting potent probiotic properties were identified as Bacillus cereus P1, Bacillus subtilis P6, B. subtilis P7, Pseudomonas aeruginosa P12 and P. aeruginosa P15 on the basis of morphological, biochemical and molecular characteristics. Maximum phytase activity (2.74 EU mL^?1) with potential probiotic properties, that is more than 70% survivability at pH 2.0, up to 2.0% bile salt tolerance, sporulation efficiency of more than 80% and survival in anaerobic condition (94.31%), was revealed by B. subtilis P6 as compared to the well‐established commercial probiotic strains Lactobacillus sporogenes and Lactobacillus casei. Thus, phytase‐producing B. subtilis P6 with promising probiotic features can be used in food and animal feed applications for betterment of mankind after further validation.     

The survival and persistence in the human gastrointestinal tract of five potential probiotic lactobacilli consumed as freeze-dried cultures or as probiotic sausage

Among five lactobacilli (L. plantarum MF1291, MF1298, DC13, L. pentosus MF1300 and L. salivarius DC5) which were administrated as freeze-dried cultures for 17 volunteers, MF1298 and DC13 were the most frequently reisolated strains in faeces demonstrating the human gastric survival of these strains. Furthermore, MF1298 and DC13 persisted in the same volunteer after ended intake, suggesting host-specific persistence behaviour. When MF1298 was administrated as sausage fermented with this strain, the number of volunteers harbouring MF1298 increased from 4 to 10 indicating that the sausage matrix protects the survival through the gastrointestinal tract (GIT).     

Quality characteristics of kefir as a carrier for probiotic Lactobacillus rhamnosus GG

Kefir was prepared using kefir grains of different origins (US, Australia and India) and starter culture. The sensory scores for kefir made using starter culture and US kefir grains were significantly higher than for the other samples (P < 0.05). The index of viscosity was significantly higher (P < 0.05) in kefir prepared using Indian kefir grains. Shear stress–shear rate plots indicated yield stress in all kefirs. All samples exhibited shear‐thinning behaviour. The average count of Lactobacillus rhamnosus GG in kefir after incubation was 7.09 log CFU/g. No significant difference (P > 0.05) was observed in quality characteristics in kefir after the addition of LGG, indicating that it could be a potential probiotic carrier.     

Meat as a functional food with special reference to probiotic sausages

Consumers believe that foods are associated directly to their health. Today foods are not only used to satisfy our hunger but also to provide indispensible nutrients for humans and these nutrients having the health benefits regarding in controlling the diseases. The market for functional foods has seen a sharp rise in demand in the recent years. This has driven researchers to multiply their efforts in producing functional meat products also. Feed manipulation and post-mortem modification of meat coupled with enrichment of bioactive compounds are gaining importance. This review discusses the candidate ingredients and strategies, utilized in crafting such functional meat products, and the notable developments and commercial successes in functional meat industry. Dry fermented sausages meet the conditions required to carry viable probiotic microbes. This article enlists various microorganisms that are being commercially used in functional food products and potential bacteria for probiotic sausage production.     

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.     

Survival of a probiotic, Lactobacillus casei strain Shirota, in the gastrointestinal tract: selective isolation from faeces and identification using monoclonal antibodies.

Lactobacillus casei strain Shirota (LCS) is a probiotic bacterium used in the production of fermented milk products and lactic acid bacteria preparations. To investigate the survival of LCS in the gastrointestinal tract, we have developed a selective medium and specific monoclonal antibodies to isolate and identify this strain. Selective LLV agar medium was prepared by modifying LBS medium, a selective medium for lactobacilli, through the replacement of glucose with lactitol as a carbon source and vancomycin as a selective antibiotic. Culture in LLV agar medium followed by ELISA using monoclonal antibodies specific for LCS was able to detect the organism in faeces. Using this method, we studied the faecal recovery of LCS in individuals who drank 125 ml of fermented milk which contained 10(10) live LCS for 3 days. The mean recovery was about 10(7) live bacteria per gram of faeces, indicating that LCS survived transit through the gastrointestinal tract after ingestion of the fermented milk.     

Pediococcus acidilactici as a potential probiotic to be used in food industry

The objective of this study was to compare the results obtained between a commercial probiotic bacterium and a strain of Pediococcus acidilactici isolated from a food matrix. Besides the characterisation of the isolates, the main focus of this work was to evaluate their ability to survive through simulated gastrointestinal tract passage as well as the effect of the spray drying process to be used as concentrated cultures in the food industry. Both lactic acid bacteria produced any of the virulence determinants investigated; they exhibited no significant antibiotic resistances and <1 log unit reduction in viable counts was obtained after exposure to simulated gastrointestinal tract conditions. Concentrated cultures of P. acidilactici HA‐6111‐2 in 10% (w/v) of reconstituted skim milk were obtained by spray drying and, after 60 days of storage at 4 °C, <1 log unit reduction in viable count was recorded. In conclusion, from these preliminary characterisation tests and its extended viability in the dried form, this P. acidilactici strain should be considered as a potentially useful probiotic.     

Production of probiotic Feta cheese using Propionibacterium freudenreichii subsp. shermanii as adjunct

Probiotic Feta cheese was produced using Propionibacterium freudenreichii subsp. shermanii as an adjunct since it has been shown to induce apoptosis of colon cancer cells in vitro and in vivo through the production of propionate and acetate. Microbiological and physicochemical analysis of the cheese was performed through the 60 d period of ripening. Counts of propionibacteria increased until day 7 and then remained constant until the end of ripening (approximately 9 log cfu g⁻¹). Moreover, throughout ripening the presence of P. freudenreichii subsp. shermanii LMG 16424^T was confirmed by 16S rRNA gene sequence analysis, while propionic acid was first detected on day 7 and reached a concentration of 52.1 mm on day 60. The ripened cheese containing P. freudenreichii subsp. shermanii was very well accepted by the sensory evaluation panellists. This is the first time that P. freudenreichii subsp. shermanii was studied as a probiotic adjunct in a white-brined cheese.