Short communicationSupercritical carbon dioxide interpolymer complexes improve survival of B. longum Bb-46 in simulated gastrointestinal fluids
Introduction
Several probiotic lactic acid bacteria strains are available to consumers in both traditional fermented foods and in supplement form (Kourkoutas et al., 2005). Numbers of viable organisms in products are reduced due to exposure of products to different stresses during manufacturing, storage and consumption (Doleyres and Lacroix, 2005). However, probiotic cultures must remain viable in the environment where they act, to enable them to exert beneficial effect on the consumer (Schillinger, 1999).
These organisms must therefore survive the journey through the upper GIT so that they reach the colon in large numbers to colonize the host (Kailasapathy and Rybka, 1997, Alander et al., 1999, Lian et al., 2003, Hsiao et al., 2004, Mainville et al., 2005). On arrival in the colon, the ingested probiotics compete with other bacterial species already present for nutrients and adherence sites on the intestinal epithelium (Alander et al., 1999). Viability of these cultures in the GIT is affected mainly by gastric acid present in the stomach and bile in the duodenum (Rao et al., 1989, Lo et al., 2004, Mainville et al., 2005). This sensitivity of probiotics presents a challenge for their application in different industries (Hansen et al., 2002).
Several studies have shown poor survival of many strains of bifidobacteria in acidity and bile concentration present in the human GIT. Approaches for improving survival of these bacteria include selection of acid and bile resistant strains, use of O2 impermeable containers, two-step fermentations, stress adaptation, incorporation of micronutrients and microencapsulation (Picot and Lacroix, 2004).
Microencapsulation of bifidobacteria for improving gastrointestinal survival has been studied by various researchers (Rao et al., 1989, Sheu and Marshall, 1993, Cui et al., 2000, Lee and Heo, 2000, Sultana et al., 2000, Sun and Griffiths, 2000, Hansen et al., 2002, Guérin et al., 2003, Lian et al., 2003, Krasaekoopt et al., 2004, Capela et al., 2006). Most results indicated improved survival. However, most of the methods present problems for large scale production though promising on a laboratory scale (Picot and Lacroix, 2004). Also, these methods typically involve exposure of the probiotics to either water or organic solvent. This may compromise survival of encapsulated cells as they are sensitive to solvents and moisture. Thus, use of solvents should be avoided in order to improve survival. None of these previous studies reported survival of probiotics encapsulated in an interpolymer complex in supercritical CO2 (scCO2). This approach was reported for the first time by this group (Moolman et al., 2006). The aim of this study was to investigate the survival of interpolymer complex encapsulated Bifidobacterium longum Bb-46 in SGF and SIF, and to investigate effects of different modifications of the polymers on bacterial survival.
Section snippets
Bacterial cultures
B. longum Bb-46 was obtained in freeze-dried form from Chr-Hansen. The culture was stored at − 20 °C and then used as freeze-dried powder in encapsulation experiments.
Polymer formulations
Different polymer formulations used for encapsulation of bacteria are summarized in Table 1.
Preparation of ingredients for encapsulation
All equipment was wiped with 70% ethanol (NCP Alcohols) using a paper towel, and allowed to dry before contact with the materials. Poly (vinylpyrrolidone) (PVP) (Kollidon 12PF, mass-average molar mass 2000–3000 g/mol, BASF) was dried for 5 h
Survival in the basic system and with added copolymer
Probiotic cultures must withstand the acidic conditions of the stomach and reach the colon in large quantities (Kailasapathy and Rybka, 1997, Alander et al., 1999, Lian et al., 2003, Hsiao et al., 2004, Mainville et al., 2005). The encapsulated probiotic bacteria were therefore exposed to SGF and SIF to investigate the potential of the encapsulation for improving survival of the bacteria under the unfavourable conditions in upper sections of the GIT. Fig. 1 shows comparative counts for
Conclusions
Encapsulation in an interpolymer complex in scCO2 improved survival of B. longum Bb-46 through a simulated gastrointestinal envionment. The encapsulation method therefore has potential for application in food and pharmaceutical industries. Future in vitro studies will investigate the effect of the encapsulated bacteria on the microflora of the simulator of the human intestinal microbial ecosystem (SHIME) model. The effect of encapsulation on the shelf life of probiotics will also be
Acknowledgements
Thanks are due to Ellipsoid Technology (Pty) Ltd and the University of Pretoria for financial support.
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