PROBIOCAPS - tailored microencaPsulation technology foR prOBIOtic baCteriA: Production, stability and functionality enhancementS in various carriers

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Characterization of freezing effect upon stability of, probiotic loaded, calcium-alginate microparticles

Publication . 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.

2015Journal article

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Storage Stability of Lactobacillus paracasei as Free Cells or Encapsulated in Alginate-Based Microcapsules in Low pH Fruit Juices

Publication . 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.

2012Journal article

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Encapsulation of probiotic strains in plain or cysteine-supplemented alginate improves viability at storage below freezing temperatures

Publication . 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.

2012Journal article

Restricted access

Optical fibre-based methodology for screening the effect of probiotic bacteria on conjugated linoleic acid (CLA) in curdled milk

Publication . Silva, Lurdes I. B.; Rodrigues, Dina M.; Freitas, Ana C.; Gomes, Ana M.; Rocha-Santos, Teresa A. P; Pereira, M. E.; Duarte, A. C.

A methodology based on optical fibre (OF) detection was developed for screening the potential of CLA production by Lactobacillus casei-01, Lactobacillus acidophilus La-5 and Bifidobacterium lactis B94 in probiotic curdled milk. The OF based methodology was validated by comparison with an analytical method based on gas chromatography-mass spectrometry (GC-MS) and it showed comparable linearity (between 5 and 130 mu g), accuracy and detection limits, which ranged from 1.92 to 2.56 mu g for CLA methyl ester and oleic acid methyl ester, respectively. Furthermore, the proposed OF based methodology was an advantageous analytical methodology for screening CIA production in probiotic curled milk, due to its compact design and effective cost of analysis. The application of the OF analytical method allowed to detect CLA in the probiotic curdled milk: the production of CLA by probiotic enzymes reached values of 297 +/- 19 mu g g(-1) by 60 days of ripening unfolding this potential functional food as very interesting from a technological point of view.

2011Journal article

Open access

Development of probiotic tablets using microparticles: viability studies and stability studies

Publication . 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.

2013Journal article

Restricted access