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- Influence of L-cysteine, oxygen and relative humidity upon survival throughout storage of probiotic bacteria in whey protein-based microcapsulesPublication . Rodrigues, D.; Sousa, Sérgio; Santos, Nathália Baptista T.; Silva, J.P.; Lobo, J. M. Sousa; Costa, P; Amaral, Gustavo Henrique De Oliveira; Pintado, M.M.E; Gomes, Ana Maria; Malcata, F. Xavier; Freitas, Ana CThe survival rates of Lactobacilus acidophilus Ki, Lactobacillus paracasei L26 and Bifidobacterium animalis BB-12 were studied after whey protein microencapsulation via spray-drying, with or without L-cysteine-HCl, and storage up to 6 months at 5 degrees C and 22 degrees C, with variation in relative air humidity and oxygen levels. Lb. paracasei L26 was the least susceptible to storage conditions: above 10(6) cfu g(-1) were recorded by 180 d at 22 degrees C, irrespective of relative humidity, and the presence/absence of oxygen and L-cysteine. Higher relative humidity, higher temperature and longer storage periods were deleterious to survival of both B. animalis BB-12 and Lb. acidophilus Ki; the effect of L-cysteine-HCl was dependent on the probiotic strain. The effect of overhead oxygen was not significant upon any probiotic strain studied. Whey protein microcapsules containing L-cysteine-HCl protected probiotic cultures from simulated gastrointestinal conditions.
- Effects of encapsulation on the viability of probiotic strains exposed to lethal conditionsPublication . Borges, Sandra; Barbosa, Joana; Camilo, Rute; Carvalheira, Ana; Silva, Joana; Sousa, Sérgio; Gomes, Ana M.; Pintado, Maria M.; Silva, José P.; Costa, Paulo; Amaral, Maria H.; Teixeira, Paula; Freitas, Ana C.The effect of microencapsulation on the viability of Lactobacillus casei, L. paracasei, L. acidophilus Ki and Bifidobacterium animalis BB-12 during exposure to lethal conditions (25% NaCl, pH 3.0 and 55–60 ºC) was evaluated. Results demonstrated that survival of probiotic strains to the imposed lethal stress conditions was strain dependent. With the exception of exposure to 25% (w ⁄ v) NaCl, L. acidophilus Ki (free and encapsulated cells) demonstrated the highest survival rates through exposure to lethal conditions of temperature and pH. For this probiotic strain exposed to heat, microencapsulated cells expressed a higher heat tolerance at 55 ºC than free cells. For the other tested bacteria, in general, encapsulation had no positive effect on survival through the tested lethal conditions.
- Effects of processing and storage on Pediococcus pentosaceus SB83 in vaginal formulations: lyophilized powder and tabletsPublication . Borges, Sandra; Costa, Paulo; Silva, Joana; Teixeira, PaulaVaginal probiotics have an important role in preventing the colonization of the vagina by pathogens. This study aimed to investigate different formulations with Pediococcus pentosaceus SB83 (lyophilized powder and tablets with and without retarding polymer) in order to verify its stability and antilisterial activity after manufacture and during storage. The bacteriocinogenic activity of P. pentosaceus SB83 against Listeria monocytogenes was evaluated in simulated vaginal fluid. Suspension of Pediococcus pentosaceus SB83 reduced the pathogen only after 2 h and the lyophilized bacteria after 24 h of contact, and, in the tablets, P. pentosaceus SB83 lost the antimicrobial activity. The pH of simulated vaginal fluid decreased for all the tested conditions. As lyophilized powder demonstrated better results concerning antimicrobial activity, this formulation was selected to evaluate the antilisterial activity during the 12 months of storage. During storage at room temperature, lyophilized bacteria totally inhibited the pathogen only until one month of storage. At 4 degrees C, P. pentosaceus SB83 showed antimicrobial activity during all the time of storage investigated. Therefore, the better formulation of P. pentosaceus SB83 is the lyophilized powder stored at 4 degrees C, which may be administered intravaginally as a washing solution.
- Development of probiotic tablets using microparticles: viability studies and stability studiesPublication . Sousa e Silva, J. P.; Sousa, Sérgio C.; Costa, Paulo; Cerdeira, Emília; Amaral, Maria H.; Lobo, José Sousa; Gomes, Ana M. P.; Pintado, Maria M.; Rodrigues, Dina; Rocha-Santos, Teresa; Freitas, A. C.Alternative vectors to deliver viable cells of probiotics, to those conferring limited resistance to gastrointestinal conditions, still need to be sought. Therefore the main goal of the study was to develop tablets able to protect entrapped probiotic bacteria from gastric acidity, thus providing an easily manufacturing scale-up dosage form to deliver probiotics to the vicinity of the human colon. Whey protein concentrate microparticles with Lactobacillus paracasei L26 were produced by spray-drying and incorporated in tablets with cellulose acetate phthalate and sodium croscarmellose. The viability of L. paracasei L. 26 throughout tableting as well as its gastric resistance and release from the tablets were evaluated. Storage stability of L. paracasei L26 tablets was also performed by evaluation of viable cells throughout 60 days at 23 degrees C and 33% relative humidity. A decrease of approximately one logarithmic cycle was observed after the acid stage and the release of L. paracasei L26 from the tablets occurred only after 4 h in the conditions tested. Microencapsulated L. paracasei L26 in tablets revealed some susceptibility to the storage conditions tested since the number of viable cells decreased 2 log cycles after 60 days of storage. However, the viability of L. paracasei L26 after 45 days of storage did not reveal significant susceptibility upon exposure to simulated gastrointestinal conditions. The developed probiotic tablets revealed to be potential vectors for delivering viable cells of L. paracasei L26 and probably other probiotics to persons/patients who might benefit from probiotic therapy.
- Encapsulation of probiotic strains in plain or cysteine-supplemented alginate improves viability at storage below freezing temperaturesPublication . Sousa, Sérgio; Gomes, Ana M.; Pintado, Maria M.; Malcata, Francisco X.; Silva, José P.; Sousa, José M.; Costa, Paulo; Amaral, Maria H.; Rodrigues, Dina; Rocha-Santos, Teresa A. P.; Freitas, Ana C.Four probiotic bacteria (Lactobacillus paracasei L26, L. casei-01, L. acidophilus Ki, and Bifidobacterium animalis BB-12 R ) were encapsulated in plain alginate or alginate supplemented with L-cysteine·HCl, and resulting microcapsules were stored at different temperatures, namely 21, 4, −20, or −80◦C for a period of up to6months. The results showed that the encapsulation in calcium alginate microcapsules was only effective in promoting protection at freezing temperatures, independently of the sensitivity of the strain. Storage of calcium alginate microcapsules at −80◦C indicated a protective effect upon viability of all four probiotic strains and the presence of L-cysteine·HCl in the alginate matrix improved protection upon cell viability of B. animalis BB-12 R . An increase in storage temperature of encapsulated bacteria caused an increase in rate of loss in their viability that was strain dependent. This study suggests that microencapsulation of probiotic cells in calcium alginate can be suitable for sustaining the viability of probiotics in food products that require storage below freezing temperatures, even in the absence of cryoprotectors, contributing to an increased shelf life.
- Characterization of freezing effect upon stability of, probiotic loaded, calcium-alginate microparticlesPublication . Sousa, Sérgio; Gomes, Ana M.; Pintado, Maria M.; Silva, José P.; Costa, Paulo; Amaral, Maria H.; Duarte, Armando C.; Rodrigues, Dina; Rocha-Santos, Teresa A. P.; Freitas, Ana C.Microencapsulation, utilizing different techniques and polymers, has been studied with the objective of maintaining probiotic viability in food matrices, protecting the cells from their detrimental environment, storage conditions andthe passage of gastrointestinal tract (GIT). The main objective of this study was to assess the effect of freezing at−20◦C upon probiotic alginate-calcium microparticles’ integrity and functionality through parameters such as size,morphology and structure of microparticles as well as to assess cell resistance to simulated gastrointestinal tractconditions upon storage. In order to study the effect of freezing upon the stability of the microparticles, calcium-alginate microparticles, with or without probiotic cells (Lactobacillus casei-01, Lactobacillus paracasei L26, Lactobacillusacidophilus KI and Bifidobacterium animalis BB-12), were characterized at production time and after 60 days storage at−20◦C. An increase in particle size, loss of the spherical shape and porous net damages were observed after 60 daysof storage at −20◦C. In accordance, encapsulation in alginate was not able to exert protection to the encapsulatedprobiotic cells stored at −20◦C for 60 days, especially from acid and particularly bile salts. B. animalis BB-12 revealedto be the most resistant probiotic strain, to both the microencapsulation process and to GIT simulated conditions.
- On the viability of five probiotic strains when immobilized on various polymersPublication . Rodrigues, Dina; Rocha-Santos, Teresa; Sousa, Sérgio; Gomes, Ana M; Pintado, M. E.; Malcata, F. Xavier; Lobo, José M. Sousa; Silva, José P.; Costa, Paulo; Amaral, Maria H.; Freitas, Ana C.The viability of probiotic bacteria in six food-grade polymers, at two concentrations, was evaluated in order to predict their feasibility as materials for bacterium immobilisation. Alginate and whey proteins were the most adequate polymers, except for Lactobacillus acidophilus Ki and Lactobacillus casei 01 at 2% (m/v) alginate. Xanthan gum appeared to be a potential vector for three strains. L-carrageenan was adequate for both Bifidobacterium strains, but not for Lactobacillus at 2% (m/v). Bifidobacterium strains were not negatively affected by cellulose acetate phthalate, while the opposite held for L. acidophilus strains. Chitosan was the poorest polymer for immobilisation of probiotic bacteria.
- Storage Stability of Lactobacillus paracasei as Free Cells or Encapsulated in Alginate-Based Microcapsules in Low pH Fruit JuicesPublication . Rodrigues, Dina; Sousa, Sérgio; Gomes, Ana M.; Pintado, Maria M.; Silva, José P.; Costa, Paulo; Amaral, Maria H.; Rocha-Santos, Teresa; Freitas, Ana C.The main objective of this research effort was to study whether microencapsulation could be a viable alternative to obtain probiotic orange or peach juices. In order to be considered probiotic food, probiotic bacteria must be present in sufficient viable numbers to promote a benefit to the host. The survival and viability of Lactobacillus paracasei L26 in juices over 50 days of storage at 5°C was assessed, evaluating the potential use of encapsulated cells in alginate microcapsules. L. paracasei L26 demonstrated good viability in both orange and peach juices despite the low pH values of both juices. Microencapsulation in alginate, with or without double coating, revealed to be suitable to protect L. paracasei L26 since viable cells were approximately 9 log cfu/g after 50 days of storage at 5°C. In general, the probiotic fruit juices showed a decrease in pH during storage. Glucose and fructose contents as well as citric acid contents decreased during storage, whereas an increase in formic acid was observed. The outcome of this study points to L. paracasei L26 as having promising potential, especially in an encapsulated form, as functional supplements in fruit juices without dairy ingredients due to their tolerance in an acidic environment over 50 days of storage at 5°C. Further studies are warranted to prove the functionality of juices with encapsulated probiotic strains.