CONICET | Buscador de Institutos y Recursos Humanos

Introduction Probiotics are live microorganisms that administered in adequate amounts, 106-107 CFU/mg, confer a beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 6-107 CFU/mg, confer a beneficial effect on the health of the host, being able to be used for therapeutic purposes.1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 1 However, these bacteria are sensitive to gastric secretions and bile salts. Therefore, is necessary to develop systems capable of protecting them. Cell microencapsulation in alginate matrix has been widely used to improve their viability in gastric medium.2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 biopolymers has been not investigated deeply.3 2 However, the protection of probiotics using alginate mixed with other biopolymers has been not investigated deeply.33 Materials and Methods Microencapsulation methodology: sodium alginate and each of the polymers (CarbopolR, carboxymethylcellulose, pregelatinized starch, chitosan), were dissolved in distilled water. The encapsulated Lactococcus lactis NZ9000 were grown in liquid medium (M17). The cells were centrifuged and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.3 and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.3 and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.3 and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.3 and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.3 Lactococcus lactis NZ9000 were grown in liquid medium (M17). The cells were centrifuged and resuspended with sodium alginate solution and the polymer. The suspension formed drip was added to a solution of calcium chloride 3% with stirring yielding alginate microparticles with encapsulated bacteria. The solid was filtered and washed with sterile distilled water and dried at 30 °C.33 Encapsulation test: The microparticles were dissolved in buffer pH 7.40. It took an aliquot of the solution and serial dilutions were made, sowed on plates with half Agar M17, incubated 24 hours at 30 °C and then came the plate count. Tolerance acid and bile salts test: The microparticles were incubated 2 hours at pH 1.5. Another batch of microparticles was incubated for 2 hours in a solution of 0.6% bile salts pH 7.4. An aliquot of each batch and dilutions were planted in half M17-Agar (24 hs at 30 °C) and then counting the colonies, expressing the results as CFU / g of microparticles.44 Release rate of bacteria: The microparticles were dissolved in buffer pH 7.4 and aliquots were taken at different times, appropriate dilutions were made, sowed on plates with half Agar M17, and then counting the number of colonies after 24 hours incubation at 30 oC. Results High cell loading (> 70%) was achieved by formulating alginate with carboxymethylcelullose. Survival of the microorganisms was found to increase on alginate-carboxymethylcelluose in comparison with free cells, and alginate pure encapsulation at different conditions of pH and bile salts. CarbopolR, chitosan, and pregelatinized starch did not show any advantanges over pure alginate microcapsules on the viability of the cells. In size and shape the microparticles, characterized by light and electron microscopy, we observed that ranged in size from 550-650 microns. Conclusion Microencapsulated cells of Lactococcus lactis NZ9000 in alginate beads formulated with cellulose derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. Lactococcus lactis NZ9000 in alginate beads formulated with cellulose derivatives resulted in better survived than for free cells after sequential incubation in simulated gastric, intestinal and bile salts fluids. References 1. Guarner, F., Schaafsma, G. J. International Journal of Food Microbiology, 39, 237-238, 1998. 2. Krasaekoopt, W., Bhandari, B., Deeth, H. International Dairy Journal, 13, 3-13, 2003. 3. Vandenberg, G.W., Drolet, C.,Scott1,, S.L., de la Noue, J. Journal of Controlled Release 77, 297-307, 2001. 4. Ding, W.K., Shah, N.P. Journal of Food Science, 72, M446-M450, 2007