Shell-Core Microbeads Loaded with Probiotics: Influence of Lipid Melting Point on Probiotic Activity.

Probiotics have many beneficial physiological activities, but the poor stability during storage and gastrointestinal digestion limits their application. Therefore, in this study, a novel type of shell-core microbead for loading probiotics was prepared through high-precision concentric drop formation technology using gelatin as the shell material and lipids as the core material. The microbeads have a regular spherical structure, uniform size, low moisture content (<4%) and high probiotic activity (>9.0 log CFU/g). Textural testing showed that the hardness of the medium-chain triglyceride microbeads (MCTBs), cocoa butter replacer microbeads (CBRBs) and hydrogenated palm oil microbeads (HPOBs) increased gradually (319.65, 623.54, 711.41 g), but their springiness decreased (67.7, 43.3, 34.0%). Importantly, lipids with higher melting points contributed to the enhanced stability of probiotics during simulated digestion and storage. The viable probiotic counts of the HCTBs, CBRBs and HPOBs after being stored at 25 °C for 12 months were 8.01, 8.44, and 8.51 log CFU/g, respectively. In the simulated in vitro digestion process, the HPOBs resisted the destructive effects of digestive enzymes and gastric acid on probiotics, with a reduction in the probiotic viability of less than 1.5 log CFU/g. This study can provide new ideas for the preparation of intestinal delivery probiotic foods.

Evaluation of the Functional and Sensory Properties of Probiotic Whey Cheese with Herbs in Vacuum or Modified Atmosphere Packaging.

Research background: Some herbs provide functional properties to foods, especially their antibacterial and antioxidant properties. On the other hand, modified atmosphere packaging is being considered as an alternative to vacuum packaging to preserve the functional and sensory properties of foods. Since the shelf life of whey cheese is quite short, different packaging methods such as modified atmosphere packaging are favoured. Besides, the addition of herbs both gives flavour to the cheese and improves its functional properties. Experimental approach: In the present study, oregano (Origanum onites) or rosemary (Rosmarinus officinalis) was added to probiotic whey cheese (lor) containing Lactobacillus acidophilus La-5 and Bifidobacterium lactis Bb-12 under modified atmosphere packaging (MAP) (80 % CO2 and 20 % N2) or vacuum packaging. The physicochemical, microbiological and sensory properties as well as antioxidant and proteolytic activities of the cheese samples were determined. Results and conclusions: The addition of herbs did not negatively affect the viable counts of B. lactis and L. acidophilus, and the cheese samples contained at least 8 log CFU/g of both probiotic bacteria for 35 days. MAP improved the viability of B. lactis and L. acidophilus in cheese with rosemary during the first few weeks of storage compared to vacuum packaging. The addition of herbs significantly increased the total phenolic content and antioxidant activity under both MAP and vacuum. MAP improved the antioxidant activity of lor cheese with added herbs on days 14 and 28 more than vacuum packaging. Lor cheese with rosemary under MAP conditions showed the highest DPPH˙ (2,2,-diphenyl-1-picrylhydrazyl) scavenging activity and also the highest proteolytic activity throughout storage. The sample with rosemary under MAP had the highest taste and aroma scores throughout the entire storage period. Fortification with herb and MAP offers advantages in the production of whey cheese. The use of rosemary and modified atmosphere packaging makes it possible to achieve high viability of probiotic bacteria, total phenolic content, antioxidant activity and sensory acceptance in lor cheese. Novelty and scientific contribution: This is the first study in which both different herbs and different packaging methods were applied to probiotic whey cheese (lor). The study shows that the functional properties of whey cheese can be improved by using different herbs under different packaging conditions. Among the analysed properties of the product, the improvement of the viability of probiotic bacteria is particularly valuable for human health. Thus, it contributes to the science of functional food and enables the use of these parameters in some other foods.

Improving the physicochemical and antioxidative properties of fermented goat milk using carob molasses and some probiotic strains.

Fermented goat milk samples, processed by adding 0, 2, and 4% carob molasses, and Bifidobacterium bifidum DSMZ or Lactobacillus helveticus CH5, were evaluated for their physicochemical, antioxidative, and sensory properties along with probiotic viability during storage. pH and viscosity gradually decreased over the storage period whereas their values increased with the increased amount of carob molasses. Bifidobacterium bifidum DSMZ or Lactobacillus helveticus CH5 kept higher viable numbers. Lactobacillus helveticus CH5 strain recorded higher viability. Increasing the added amount of carob molasses increased the total phenolic content, which in turn is reflected in the increase in antioxidant activity. Adding carob molasses participated in the partial masking of the goaty flavor and had no significant effect on the product's acceptability, which was strongly influenced by the progress of cold storage. Therefore, fermented goat milk described by its enhanced physicochemical and antioxidative properties could be obtained by adding carob molasses and probiotics. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01382-2.

Encapsulation of probiotic bacteria using polyelectrolytes stabilized nanoliposomes for improved viability under hostile conditions.

Probiotics viability and stability is a core challenge for the food processing industry. To prolong the viability of probiotics (Lactobacillus acidophilus), gelatin (GE)-chitosan (CH) polyelectrolytes-coated nanoliposomes were developed and characterized. The average particle size of the nanoliposomes was in the range of 131.7-431.6 nm. The mean zeta potential value of the nanoliposomes differed significantly from -42.2 to -9.1 mV. Scanning electron micrographs indicated that the nanoliposomes were well distributed and had a spherical shape with a smooth surface. The Fourier transform infrared spectra revealed that the GE-CH polyelectrolyte coating has been effectively applied on the surface of nanoliposomes and L. acidophilus cells were successfully encapsulated in the lipid-based nanocarriers. X-ray diffraction results indicated that nanoliposomes are semicrystalline and GE-CH polyelectrolyte coating had an influence on the crystalline nature of nanoliposomes. Moreover, the coating of L. acidophilus-loaded nanoliposomes with GE-CH polyelectrolytes significantly improved its viability when exposed to simulated gastrointestinal environments. The findings of the current study indicated that polyelectrolyte-coated nanoliposomes could be used as an effective carrier for the delivery of probiotics and their application to food matrix for manufacturing functional foods.

Lactobacillus reuteri-fortified camel milk infant formula: Effects of encapsulation, in vitro digestion, and storage conditions on probiotic cell viability and physicochemical characteristics of infant formula.

Lactobacillus reuteri fortified camel milk infant formula (CMIF) was produced. The effect of encapsulation in different matrices (sodium alginate and galacto-oligosaccharides) via spray drying, simulated infant gastrointestinal digestion (SIGID), and storage conditions (temperature and humidity) on the viability of L. reuteri in CMIF and the physicochemical properties of CMIF were evaluated. Compared with free cells, probiotic cell viability was significantly enhanced against SIGID conditions upon encapsulation. However, L. reuteri viability in CMIF decreased after 60 d of storage, predominantly at higher storage humidity and temperature levels. At the end of the storage period, significant changes in the color values were observed in all CMIF, with a reduction in their greenness, an increase in yellowness, and a wide variation in their whiteness. Moreover, pH values and caking behavior of all CMIF stored at higher temperature (40°C) and humidity [water activity (aw) = 0.52] levels were found to be significantly higher than the samples stored under other conditions. Over 30 d of storage at lower humidity conditions (aw = 0.11 and 0.33) and room temperature (25°C), no significant increase in CMIF lipid oxidation rates was noted. Fourier-transform infrared spectroscopy analysis showed that, compared with the other storage conditions, CMIF experienced fewer changes in functional groups when stored at aw = 0.11. Microscopic images showed typical morphological characteristics of milk powder, with round to spherical-shaped particles. Overall, camel milk fortified with encapsulated L. reuteri can be suggested as a promising alternative in infant formula industries, potentially able to maintain its physicochemical characteristics as well as viability of probiotic cells when stored at low humidity levels (aw = 0.11) and temperature (25°C), over 60 d of storage.

Value-Added Utilization of Citrus Peels in Improving Functional Properties and Probiotic Viability of Acidophilus-bifidus-thermophilus (ABT)-Type Synbiotic Yoghurt during Cold Storage.

Citrus peel, a fruit-processing waste, is a substantial source of naturally occurring health-promoting compounds, including polyphenols, and has great potential as a dietary supplement for enhancing the functional properties of food. The present work aimed to investigate the effects of sour orange (SO), sweet orange (SWO), and lemon (LO) peels on the typical physiochemical, antioxidant, antibacterial, and probiotic properties of synbiotic yoghurt fermented by acidophilus-bifidus-thermophilus (ABT)-type cultures during cold storage (0−28 days). High-performance liquid chromatography-diode array detection (HPLC-DAD) analysis showed that the total phenolic content in the SO peel were more than 2-fold higher than that in the SWO and LO peel. The predominant phenolic compounds were myricetin (2.10 mg/g dry weight) and o-coumaric acid (1.13 mg/g) in SO peel, benzoic acid (0.81 mg/g) and naringin (0.72 mg/g) in SWO peel, and benzoic acid (0.76 mg/g) and quercetin (0.36 mg/g) in LO peel. Only 0.5% (w/w) of citrus peel addition did not reduce the overall acceptance of ABT synbiotic yoghurt but led to increased acidity and decreased moisture during cold storage (14 and 28 days). Additionally, compared to control samples without citrus peel addition, supplementation with citrus peels improved the antioxidant property of the ABT synbiotic yoghurt. ABT milks with SO and SWO peel addition had significantly stronger DPPH radical scavenging activities than that with LO peel addition (p < 0.05). Antibacterial analysis of ABT synbiotic yoghurt with citrus peel addition showed that the diameters of inhibition zones against S. aureus, B. subtilis, and E. coli increased by 0.6−1.9 mm relative to the control groups, suggesting the enhancement of antibacterial activities by citrus peels. The viabilities of probiotic starter cultures (L. acidophilus, S. thermophilus, and Bifidobacterial sp.) were also enhanced by the incorporation of citrus peels in synbiotic yoghurt during cold storage. Hence, our results suggest that citrus peels, especially SO and SWO peels, could be recommended as a promising multifunctional additive for the development of probiotic and synbiotic yoghurt with enhanced antioxidant and antibacterial properties, as well as probiotic viability.

Microbiological Testing of Probiotic Preparations.

Probiotic microorganisms that are potentially beneficial to the health of the host are commercially available in a great variety of products. Not all microorganism strains present in products have proven beneficial to the health properties. These products include not only foodstuffs but also dietary supplements, food for special medical purposes, medicinal products, as well as cosmetics and medical devices. These products contain from one to a dozen bacterial strains of the same or different species and sometimes also fungal strains. Since the pro-health effects of probiotics depend on a specific strain, the number of its cells in a dose, and the lack of pathogenic microorganisms, it is extremely important to control the quality of probiotics. Depending on the classification of a given product, its form, and its content of microorganisms, the correct determination of the number of microorganisms and their identification is crucial. This article describes the culture-dependent and culture-independent methods for testing the contents of probiotic microorganisms, in addition to biochemical and genetic methods of identification. The microbiological purity requirements for various product categories are also presented. Due to numerous reports on the low quality of probiotic products available on the market, it is important to standardise research methods for this group of products and to increase the frequency of inspections of these products.

Co-encapsulation of probiotics with prebiotics and their application in functional/synbiotic dairy products.

Oral administration of live probiotics along with prebiotics has been suggested with numerous beneficial effects for several conditions including certain infectious disorders, diarrheal illnesses, some inflammatory bowel diseases, and most recently, irritable bowel syndrome. Though, delivery of such viable bacteria to the host intestine is a major challenge, due to the poor survival of the ingested probiotic bacteria during the gastric transit, especially within the stomach where the pH is highly acidic. Although microencapsulation has been known as a promising approach for improving the viability of probiotics in the human digestive tract, the success rate is not satisfactory. For this reason, co-encapsulation of probiotics with probiotics has been practised as a novel alternative approach for further improvement of the oral delivery of viable probiotics toward their targeted release in the host intestine. This paper discusses the co-encapsulation technologies used for delivery of probiotics toward better stability and viability, as well the incorporation of co-encapsulated probiotics and prebiotics in functional/synbiotic dairy foods. The common encapsulation technologies (and the materials) used for this purpose, the stability and survival of co-encapsulated probiotics in the food, and the release behavior of the co-encapsulated probiotics in the gastrointestinal tract have also been explained. Most studies reported a significant improvement particularly in the viability of bacteria associated with the presence of prebiotics. Nevertheless, the previous research has mostly been carried out in the simulated digestion, meaning that future systematic research is to be carried out to investigate the efficacy of the co-encapsulation on the survival of the bacteria in the gut in vivo.

Characterization and Cell Viability of Probiotic/Prebiotics Film Based on Duck Feet Gelatin: A Novel Poultry Gelatin as a Suitable Matrix for Probiotics.

The probiotic viability, physicochemical, mechanical, barrier, and microstructure properties of synbiotic edible films (SEFs) based on duck feet gelatin (DFG) were evaluated. Four synbiotic systems were obtained by mixing four types of prebiotics, namely, dextrin, polydextrose, gum Arabic, and sago starch, with DFG to immobilize of probiotic (Lactobacillus casei ATCC). The ability of DFG to create a suitable matrix to increase probiotic viability was compared with those of other commercial gelatins in a preliminary evaluation. The DFG showed proper probiotic viability compared with other gelatins. The addition of prebiotics reduced the transparency of SEFs and increased color differentiation, uniformity, and complete coverage of probiotic cells. The estimated shelf-life of surviving bacteria in the SEFs stored at 4 and 25 °C showed that gum arabic showed the best performance and enhanced the viability of L. casei by 42% and 45%, respectively. Dextrin, polydextrose, and sago starch enhanced the viability of L. casei at 4 and 25 °C by 26% and 35%, 26% and 5%, and 20% and 5%, respectively. The prebiotics improved the physicochemical, mechanical, and barrier properties of all SEFs, except polydextrose film. The viability of L. casei can be increased with the proper selection of gelatin and prebiotics.

Effect of Fluidized Bed Drying, Matrix Constituents and Structure on the Viability of Probiotic Lactobacillus paracasei ATCC 55544 during Storage at 4 °C, 25 °C and 37 °C.

The stabilization of probiotics for application in non-refrigerated food products is a challenging task. In the present study, probiotic Lactobacillus paracasei (Lacticaseibacillus paracasei) ATCC 55544 cells were immobilized in a dairy matrix comprising of whole milk powder, skim milk powder, or milk protein isolate using fluidized bed drying technology. The samples were taken out at different drying stages, with an apparent water activity (aw) of aw 0.5, aw 0.4, and aw 0.3, respectively, and vacuum-packed to maintain the aw and stored at three different temperatures of 4 °C, 25 °C, and 37 °C. The study evaluated the impact of matrix constituents, milk fat, protein, and carbohydrate on the viability of encapsulated probiotic L . paracasei ATCC 55544 during storage for 1 month. The whole milk powder matrix provided superior protection to the bacteria. Confocal Laser Scanning Microscopy (CLSM) was used to investigate the structure of the immobilizing matrix and the location of the probiotic L. paracasei cells embedded within the matrix. The CLSM study revealed that the probiotic bacterial cells are mostly embedded as clusters beneath the top layer. We hypothesize that the biofilm-like structure, together with the protective whole milk powder matrix, helps to retain the superior viability of probiotic cells during storage at non-refrigerated storage conditions of 25 °C and 37 °C.

Physicochemical properties, texture, and probiotic survivability of oat-based yogurt using aquafaba as a gelling agent.

Despite high consumer demands, the manufacture of nondairy yogurt from oat milk is currently hindered due to the lack of consistency and texture. An oat-based yogurt was developed using oat milk and probiotics (Streptococcus thermophilus and Lactobacillus bulgaricus) with aquafaba (AF) and vegetable oil (VO) as added ingredients. Physicochemical analyses and viability of probiotics were investigated after yogurt formation and for 3 weeks under refrigerated storage. Results showed that adding AF decreased syneresis and increased water holding capacity during storage. Both AF and VO had a beneficial effect on hardness, the most important textural property of yogurt. Confocal laser scanning microscopy revealed that the added ingredients played a major role in the formation of the gel network structure of the yogurt. Both Streptococcus thermophilus and Lactobacillus bulgaricus remained at acceptable levels > 8.28 Log CFU/g and > 5.79 Log CFU/g after 3 weeks at 4°C regardless of the added ingredients.

Survival and storage stability of encapsulated probiotic under simulated digestion conditions and on dried apple snacks.

The objective of the current study was to explore the probiotics carrier potential of apple dried snacks and improve the survival of probiotics under simulated gastrointestinal conditions. Purposely, the probiotics were encapsulated using two hydrogel materials (sodium alginate and carrageenan) by using encapsulator. Briefly, slices of apple were immersed in solution containing free and encapsulated probiotics and then dried by conventional drying method. The dried apple snack was analyzed for different characteristics (physiochemical and microbiological) during storage. The viability of the free and encapsulated probiotics was accessed in apple snack and under simulated gastrointestinal conditions. Apple snack rich with encapsulated probiotics showed a significant result (p < .05) regarding the survival and stability. The encapsulated probiotics decreased from 9.5 log CFU/g to 8.83 log CFU/g as compared to free probiotics that decreased to 5.28 log CFU/g. Furthermore, encapsulated probiotics exhibited a better stability under simulated gastrointestinal conditions as compared to free. During storage, an increase in phenolic content and hardness was observed while decrease in pH was noted. Results of sensory parameters indicated apple snack as potential and acceptable probiotics carrier.

Production and characterization of synbiotic Doogh, a yogurt-based Iranian drink by gum arabic, ginger extract and B. bifidum.

The objective of this study was to determine the effects of using ginger extract and gum arabic as a prebiotic on the viability of Bifidobacterium bifidum and physico chemical properties of Doogh (yogurt drink) during storage at 4 °C for 30 days. The results showed that by increasing the amounts of gum arabic to 0.5%, B. bifidum count increased significantly (P < 0.05), but further increasing of the gum arabic, had no significant effect on B. bifidum count (P > 0.05). By increasing the amounts of gum arabic, the viscosity of the Doogh samples was increased and phase separation was reduced significantly (P < 0.05). The Doogh sample containing 0.25% gum arabic and 0.25% ginger extract had the highest number of probiotic population. Results showed that addition of B. bifidum increased acid development (°D) in Doogh samples during storage. In addition, phase separation in Doogh enriched by gum arabic was slower compared to control sample and the samples containing ginger extract. Doogh samples containing 0.25-1% gum arabic alone showed acceptable stability during storage time at 4 °C. In general, Doogh sample containing 0.25% ginger extract and 0.5% gum arabic gained the highest overall acceptability score in comparison with the other samples.

Incorporation of probiotics (Bifidobacterium animalis subsp. Lactis) into 3D printed mashed potatoes: Effects of variables on the viability.

3D printing is an emerging technology with the potential to revolutionize people's eating habits. This study firstly optimized the mashed potatoes (MP) formulation and correlated its 3D performance with rheological properties. Yield stress and consistency index (K) were closely related with MP's extrusion behavior, and too high values of them (like 2558 Pa and 2794 Pa·sn) caused the difficulty in extrusion process. Yield stress and elastic modulus (G') were critical to MP's self-supporting performance and too low values of them resulted in the deformation of printed parts during storage. The feasibility of incorporation of probiotics (Bifidobacterium animalis subsp. Lactis BB-12) into 3D printed mashed potatoes (MP) was then studied. MP with probiotics was printed with different nozzle diameter (0.6, 1.0 and 1.4 mm), printing temperature (25, 35, 45 and 55 °C) and evaluated for survival during extrusion and storage at 5 °C. It was found only the small nozzle diameter (0.6 mm) resulted in the reduction of probiotic viability from 9.93 log CFU/g to 9.74 log CFU/g. Greater reduction of viable counts of probiotics (from 10.07 log CFU/g to 7.99 log CFU/g) was found when the MP was held in a heating nozzle barrel at 55 °C for 45 min. No significant difference of probiotics viability in 3D printed samples was found during 12-day storage period at 5 °C. This study provides a new dimension on the development of functional foods by 3D printing.

Probiotics in Goat Milk Products: Delivery Capacity and Ability to Improve Sensory Attributes.

Dairy foods, particularly those of bovine origin, are the predominant vehicles for delivery of probiotic bacteria. Caprine (goat) milk also possesses potential for successful delivery of probiotics, and despite its less appealing flavor in some products, the use of goat milk as a probiotic carrier has rapidly increased over the last decade. This review reports on the diversity, applicability, and potential of using probiotics to enhance the sensory properties of goat milk and goat milk-based products. A brief conceptual introduction to probiotic microorganisms is followed by an account of the unique physicochemical, nutritive, and beneficial aspects of goat milk, emphasizing its advantages as a probiotic carrier. The sensory properties of probiotic-enriched goat milk products are also discussed. The maintenance of probiotic viability and desirable physicochemical characteristics in goat milk products over shelf life is possible. However, the unpleasant sensory features of some goat milk products remain a major disadvantage that hinder its wider utilization. Nevertheless, certain measures such as fortification with selected probiotic strains, inclusion of fruit pulps and popular flavor compounds, and production of commonly consumed tailor-made goat milk-based products have potential to overcome this limitation. In particular, certain probiotic bacteria release volatile compounds as a result of their metabolism, which are known to play a major role in the aroma profile and sensory aspects of the final products.

Characterization of Diversity and Probiotic Efficiency of the Autochthonous Lactic Acid Bacteria in the Fermentation of Selected Raw Fruit and Vegetable Juices.

The diversity of indigenous lactic acid bacteria (LAB) in fermented broccoli, cherry, ginger, white radish, and white-fleshed pitaya juices was analyzed using culture-independent and -dependent approaches. The major properties of selected probiotic strains, including dynamic variations in pH, viable cell counts, antibiotic resistance, bacterial adhesion to hydrophobic compounds, and survivability during simulated gastrointestinal transit, were investigated using broccoli as the fermentation substrate. In broccoli and ginger juices, the genus Lactobacillus occupied the dominant position (abundances of 79.0 and 30.3%, respectively); in cherry and radish juices, Weissella occupied the dominant position (abundances of 78.3 and 83.2%, respectively); and in pitaya juice, Streptococcus and Lactococcus occupied the dominant positions (52.2 and 37.0%, respectively). Leuconostoc mesenteroides, Weissella cibaria/soli/confusa, Enterococcus gallinarum/durans/hirae, Pediococcus pentosaceus, Bacillus coagulans, and Lactococcus garvieae/lactis subspecies were identified by partial 16S rRNA gene sequencing. In general, the selected autochthonous LAB isolates displayed no significant differences in comparison with commercial strains with regard to growth rates or acidification in fermented broccoli juice. Among all the isolates, L. mesenteroides B4-25 exhibited the highest antibiotic resistance profile (equal to that of L. plantarum CICC20265), and suitable adhesion properties (adhesion of 13.4 ± 5.2% ∼ 36.4 ± 3.2% and 21.6 ± 1.4% ∼ 69.6 ± 2.3% to ethyl acetate and xylene, respectively). Furthermore, P. pentosaceus Ca-4 and L. mesenteroides B-25 featured the highest survival rates (22.4 ± 2.6 and 21.2 ± 1.4%, respectively), after simulated gastrointestinal transit. These results indicated a high level of diversity among the autochthonous bacterial community in fermented fruit and vegetable juices, and demonstrated the potential of these candidate probiotics for applications in fermentation.

Enrichment of probiotic ice cream with different dietary fibers: Structural characteristics and culture viability.

This study evaluated the effect of 5 dietary fibers (apple, orange, oat, bamboo, and wheat) on the physicochemical, rheological, and textural characteristics; sensory properties; and culture viability of probiotic ice cream stored at -18°C for 180 d. The presence of orange and apple fibers increased the titratable acidity, decreased the lightness (color) value of the ice creams, and enhanced the red and yellow coloration. Compared with the control sample, the consistency indices and apparent viscosities of the experimental samples increased with the addition of all dietary fibers except oat fiber. The highest viscosity was obtained in the sample fortified with apple fiber, whereas the ice cream containing orange fiber showed the highest hardness after d 60 of storage. The addition of orange and apple fibers significantly increased melting resistance; however, panelists did not generally like these samples in terms of taste-flavor. All ice creams had viable counts of Lactobacillus acidophilus of ≥7 log cfu/g during storage except the samples with orange and bamboo fiber. Bifidobacterium lactis counts were also found to be >6 log cfu/g in those samples until d 150 of storage.

Bifidogenic characteristic and protective effect of saba starch on survival of Lactobacillus plantarum CIF17AN2 during vacuum-drying and storage.

Resistant starch (RS) from unripe saba banana (Musa sapientum (Linn)) (Kluai Hin) exhibited high resistance to gastric acid and intestinal amylases. Its bifidogenic effect under competition of human fecal microflora was determined in the simulated proximal region of human colon. In addition, saba RS effectively protected Lactobacillus plantarum CIF17AN2 during drying process. The maximum survival of 85.81% was achieved under vacuum drying operated at 37 °C when saba RS was added. The addition of saba RS to formulate a synbiotic product was able to retain high viability of the vacuum-dried L. plantarum during 8-week storage at ambient temperature. This is because saba RS can stabilize the moisture content of the synbiotic product. In contrast, the dramatic increase of moisture content in the vacuum-dried L. plantarum without saba RS led to significant decrease in cell survival. Moreover, saba RS could potentially protect the vacuum-dried L. plantarum from gastric acid and bile exposures.

Improved probiotic viability in stress environments with post-culture of alginate-chitosan microencapsulated low density cells.

In this study, probiotics (Saccharomyces cerevisiae Y235) were entrapped in alginate-chitosan microcapsules by emulsification/internal gelation technique. Two different encapsulation patterns were established as directly entrapped high density cells (dEHDC) and entrapped low density cells with culture (ELDCwc). The performance of microencapsulated cells, with free cells (FC) as control, was investigated against sequential stress environments of freeze-drying, storage, and simulated gastrointestinal fluids. After being freeze-dried without cryoprotectant, the survival rate of ELDCwc (14.33%) was significantly higher than 10.00% of dEHDC, and 0.05% of FC. The lower temperature (-20°C) and ELDCwc pattern were beneficial for keeping viable cells at 7.00 logCFU g(-1) after 6 months. Furthermore, the ELDCwc microcapsule maintained viable cells of 6.29 logCFU g(-1) after incubation in SGF and SIF. These studies demonstrated that the pattern of entrapped low density cells with culture was an effective and superior technique of resisting harmful stress environments.

Assessment of Probiotic Viability during Cheddar Cheese Manufacture and Ripening Using Propidium Monoazide-PCR Quantification.

The use of a suitable food carrier such as cheese could significantly enhance probiotic viability during storage. The main goal of this study was to assess viability of commercial probiotic strains during Cheddar cheesemaking and ripening (4-6 months) by comparing the efficiency of microbiological and molecular approaches. Molecular methods such as quantitative PCR (qPCR) allow bacterial quantification, and DNA-blocking molecules such as propidium monoazide (PMA) select only the living cells' DNA. Cheese samples were manufactured with a lactococci starter and with one of three probiotic strains (Bifidobacterium animalis subsp. lactis BB-12, Lactobacillus rhamnosus RO011, or Lactobacillus helveticus RO052) or a mixed culture containing B. animalis subsp. lactis BB-12 and L. helveticus RO052 (MC1), both lactobacilli strains (MC2), or all three strains (MC3). DNA extractions were then carried out on PMA-treated and non-treated cell pellets in order to assess PMA treatment efficiency, followed by quantification using the 16S rRNA gene, the elongation factor Tu gene (tuf) or the transaldolase gene (tal). Results with intact/dead ratios of bacteria showed that PMA-treated cheese samples had a significantly lower bacterial count than non-treated DNA samples (P < 0.005), confirming that PMA did eliminate dead bacteria from PCR quantification. For both quantification methods, the addition of probiotic strains seemed to accelerate the loss of lactococci viability in comparison to control cheese samples, especially when L. helveticus RO052 was added. Viability of all three probiotic strains was also significantly reduced in mixed culture cheese samples (P < 0.0001), B. animalis subsp. lactis BB-12 being the most sensitive to the presence of other strains. However, all probiotic strains did retain their viability (log 9 cfu/g of cheese) throughout ripening. This study was successful in monitoring living probiotic species in Cheddar cheese samples through PMA-qPCR.