Impact of a yogurt matrix and cell microencapsulation on the survival of Lactobacillus reuteri in three in vitro gastric digestion procedures.

The goal of this study was to assess the interaction between microencapsulation and a yogurt food matrix on the survival of Lactobacillus reuteri NCIMB 30242 in four different in vitro systems that simulate a gastric environment. The four systems were: United States Pharmacopeia (USP) solutions, a static two-step (STS) procedure which included simulated food ingredients, a constantly dynamic digestion procedure (IViDiS), as well a multi-step dynamic digestion scheme (S'IViDiS). The pH profiles of the various procedures varied between systems with acidity levels being: USP > STS > IViDiS = S'IVIDiS. Addition of a food matrix increased the pH in all systems except for the USP methodology. Microencapsulation in alginate-based gels was effective in protecting the cells in model solutions when no food ingredients were present. The stability of the probiotic culture in the in vitro gastric environments was enhanced when (1) yoghurt or simulated food ingredient were present in the medium in sufficient quantity, (2) pH was higher. The procedure-comparison data of this study will be helpful in interpreting the literature with respect to viable counts of probiotics obtained from different static or dynamic in vitro gastric systems.

Carbon footprint of Canadian dairy products: calculations and issues.

The Canadian dairy sector is a major industry with about 1 million cows. This industry emits about 20% of the total greenhouse gas (GHG) emissions from the main livestock sectors (beef, dairy, swine, and poultry). In 2006, the Canadian dairy herd produced about 7.7 Mt of raw milk, resulting in about 4.4 Mt of dairy products (notably 64% fluid milk and 12% cheese). An integrated cradle-to-gate model (field to processing plant) has been developed to estimate the carbon footprint (CF) of 11 Canadian dairy products. The on-farm part of the model is the Unified Livestock Industry and Crop Emissions Estimation System (ULICEES). It considers all GHG emissions associated with livestock production but, for this study, it was run for the dairy sector specifically. Off-farm GHG emissions were estimated using the Canadian Food Carbon Footprint calculator, (cafoo)(2)-milk. It considers GHG emissions from the farm gate to the exit gate of the processing plants. The CF of the raw milk has been found lower in western provinces [0.93 kg of CO2 equivalents (CO2e)/L of milk] than in eastern provinces (1.12 kg of CO2e/L of milk) because of differences in climate conditions and dairy herd management. Most of the CF estimates of dairy products ranged between 1 and 3 kg of CO2e/kg of product. Three products were, however, significantly higher: cheese (5.3 kg of CO2e/kg), butter (7.3 kg of CO2e/kg), and milk powder (10.1 kg of CO2e/kg). The CF results depend on the milk volume needed, the co-product allocation process (based on milk solids content), and the amount of energy used to manufacture each product. The GHG emissions per kilogram of protein ranged from 13 to 40 kg of CO2e. Two products had higher values: cream and sour cream, at 83 and 78 kg of CO2e/kg, respectively. Finally, the highest CF value was for butter, at about 730 kg of CO2e/kg. This extremely high value is due to the fact that the intensity indicator per kilogram of product is high and that butter is almost exclusively fat. Protein content is often used to compare the CF of products; however, this study demonstrates that the use of a common food component is not suitable as a comparison unit in some cases. Functionality has to be considered too, but it might be insufficient for food product labeling because different reporting units (adapted to a specific food product) will be used, and the resulting confusion could lead consumers to lose confidence in such labeling. Therefore, simple units might not be ideal and a more comprehensive approach will likely have to be developed.

The impact of meals on a probiotic during transit through a model of the human upper gastrointestinal tract.

Commercial literature on various probiotic products suggests that they can be taken before meals, during meals or after meals or even without meals. This has led to serious confusion for the industry and the consumer. The objective of our study was to examine the impact of the time of administration with respect to mealtime and the impact of the buffering capacity of the food on the survival of probiotic microbes during gastrointestinal transit. We used an in vitro Digestive System (IViDiS) model of the upper gastrointestinal tract to examine the survival of a commercial multi-strain probiotic, ProtecFlor®. This product, in a capsule form, contains four different microbes: two lactobacilli (Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011), Bifidobacterium longum R0175 and Saccharomyces cerevisiae boulardii. Enumeration during and after transit of the stomach and duodenal models showed that survival of all the bacteria in the product was best when given with a meal or 30 minutes before a meal (cooked oatmeal with milk). Probiotics given 30 minutes after the meal did not survive in high numbers. Survival in milk with 1% milk fat and oatmeal-milk gruel were significantly better than apple juice or spring water. S. boulardii was not affected by time of meal or the buffering capacity of the meal. The protein content of the meal was probably not as important for the survival of the bacteria as the fat content. We conclude that ideally, non-enteric coated bacterial probiotic products should be taken with or just prior to a meal containing some fats.

A dynamic model that simulates the human upper gastrointestinal tract for the study of probiotics.

A dynamic model of the human upper gastrointestinal (GI) tract was designed to better simulate conditions of ingestion and digestion, by including a food matrix as part of the model design. The dynamic model consisted of two reactors maintained at 37 degrees C, one simulating stomach conditions and the other simulating duodenum conditions. The model was tested by comparing survival of bacteria isolated from humans (Bifidobacterium infantis, Lactobacillus johnsonii, Lactobacillus rhamnosus, and Lactobacillus acidophilus) animals (Bifidobacterium animalis, 2 strains), and fermented dairy products (Bifidobacterium longum, Lactobacillus kefir, Lactobacillus kefirgranum, and Leuconostoc mesenteroides) with their survival as determined by conventional methods. Five strains were not able to survive (>3 log reduction) 15 min in a medium acidified at pH 2.0 using the conventional testing method, but survival was improved significantly for some strains in the dynamic model. Two strains (Bifidobacterium animalis ATCC 25527 and Lactobacillus johnsonii La-1 NCC 533) showed good survival with both methods. The dynamic model was shown to better represent the events during upper GI tract transit than the conventional methods, by incorporation of a food matrix to buffer the gastric acidity and therefore expose bacteria to pH levels found in vivo before, during, and after a meal.

Microentrapment of probiotic bacteria in a Ca(2+)-induced whey protein gel and effects on their viability in a dynamic gastro-intestinal model.

Entrapping probiotic bacteria in gels with ionic cross-linking is typically achieved with polysaccharides (alginate, pectin, carraghenan). In this study, whey proteins were used for this purpose by carrying out the Ca(2+)-induced gelation of pre-heated whey protein isolate (WPI). A Lactobacillus rhamnosus cell suspension was added in a denatured WPI solution in a 30 : 70 volume ratio. Gelation was carried out by extrusion of the cell suspension in a CaCl(2) solution. Beads of approximately 3 mm diameter were formed. The population in the beads was 8.0 x 10(8) cells g(-1). Entrapment efficiency in gel beads was 96%, with a survival level of 23%. Scanning electron microscopy of beads before freeze-drying showed a tight protein network containing encapsulated Lb. rhamnosus cells homogeneously distributed throughout the matrix. The survival to freeze-drying of the bead-entrapped cells was 41%. Viability of microentrapped cells in a dynamic gastro-intestinal (GI) model was studied and the results were compared to free cells freeze-dried in a milk-based cryoprotective solution, as well as in a pre-denatured WPI solution. Results showed that protein gelation provided protection against acidic conditions in the stomach after 90 min, as well as against bile after 30, 60 and 90 min in the duodenum. Moreover, the milk-based cryoprotective solution was equally effective after 90 min in the duodenum. It is concluded that the gelation of whey proteins induced by Ca(2+) ions can protect the cells against adverse conditions of the GI system. However, certain stages in the entrapment process, particularly extrusion in the solution of CaCl(2), still need to be optimized in order to reduce the mortality of the cells during gelation.

Plasmid stability in immobilized and free recombinant Escherichia coli JM105(pKK223-200): importance of oxygen diffusion, growth rate, and plasmid copy number.

Stability of the plasmid pKK223-200 in Escherichia coli JM105 was studied for both free and immobilized cells during continuous culture. The relationship between plasmid copy number, xylanase activity, which was coded for by the plasmid, and growth rate and culture conditions involved complex interactions which determined the plasmid stability. Generally, the plasmid stability was enhanced in cultured immobilized cells compared with free-cell cultures. This stability was associated with modified plasmid copy number, depending on the media used. Hypotheses are presented concerning the different plasmid instability kinetics observed in free-cell cultures which involve the antagonistic effects of plasmid copy number and plasmid presence on the plasmid-bearing/plasmid-free cell growth rate ratio. Both diffusional limitation in carrageenan gel beads, which is described in Theoretical Analysis of Immobilized-Cell Growth, and compartmentalized growth of immobilized cells are proposed to explain plasmid stability in immobilized cells.