Dietary supplements containing viable probiotic bacteria are increasing in popularity in the marketplace as their health benefits become recognized. Reported benefits range from alleviating constipation and diarrhea to reducing various intestinal infections such as those caused by rotaviruses, pathogenic E. coli and Helicobacter pylori as discussed in U.S. Pat. Nos. 7,090,840 and 7,029,669. Beneficial species of Lactobacillus and bifidobacteria are among the widely recognized probiotics, and strains of these bacteria that are capable of colonizing the intestinal tract are advantageous (see U.S. Pat. Nos. 7,150,986 and 6,887,465).
Colonization may involve physical attachment to epithelial cell surfaces such as those of the microvilli in the ileum section of the small intestine, or simple domination of the contents of the cecum, or adherence within the mucin layers of the colon. There is much to learn about how probiotics colonize the mammalian intestinal tract but it has been established that when colonization occurs, more probiotic microorganisms appear in the feces and this correlates with more probiotic microorganisms in both the proximal and distal sections of the intestinal tract. See Muralidhara et al, 1997, Journal of Food Protection Vol. 40, No. 5, Pages 288-295. Young pigs were used to demonstrate the relationship between colonization and fecal probiotic counts in Muralidhara's work; the porcine intestinal tract is very similar in physiology to the human intestinal tract and similar studies with young pigs are presented in examples of the present invention to demonstrate its effectiveness.
Colonization that results in the competitive exclusion of pathogenic microorganisms is particularly beneficial and can occur when probiotics occupy most of the intestinal attachment sites and are encouraged to produce lactic acid and other antimicrobial compounds. Effective intestinal colonization by probiotics depends on the availability of proper microbial nutrition that must be provided by the diet. See Gibson et al 1995, Gastroenterology 106: 975-982; Christl et al, 1992, Gut 33: 1234-1238 and Reid et al, 1990, Clin. Microbiol. Rev. 3: 335-344. However, normal diets do not provide nutrients that necessarily benefit probiotics so there is a need to fortify the diet with such nutrients. Prebiotics are one class of microbial nutrients that are currently popular in the marketplace. They are typically certain oligosaccharides that are not digested in the small intestine but serve as nutrients for select probiotic bacterial genera, e.g., bifidobacteria, when they arrive in the colon. An article titled “Prebiotics Enhance Gut Health” by Laura Brandt is available online at the nutrasolutions website; another titled “Prebiotics: A More Reliable Way to Increase Gut-Friendly Bacteria” by Dr. James Meschino is available at the chiroweb website, in the archival section. Today, a variety of functional foods are fortified with prebiotics such as fructooligosaccharides (FOS) and inulin, in an effort to provide probiotic stimulation in vivo. This practice adds cost to the foods being fortified and is not very effective in stimulating Lactobacillus probiotics. Thus, there is a need for reducing the cost of prebiotics while providing for both Lactobacillus and Bifidobacterium stimulation.
Although enzymes have been used to generate prebiotics under laboratory conditions followed by subsequent feeding of the preformed prebiotics to achieve probiotic stimulation (see U.S. Pat. Nos. 6,791,015 and 6,730,502), no one has suggested using enzymes to generate these effects in vivo. U.S. Pat. No. 5,817,350 discloses the use of the prebiotic enzymes cellulose, amylase and hemicellulase, for use as dietary supplements, but not use of these enzymes to stimulate administered probiotics, or enhancement of their prebiotic effect by addition of polysorbate compounds. U.S. patent application 20010031276 discloses the use of polysorbate compounds as feed additives for ruminant animals but does not relate to their use in combination with prebiotics and probiotics, or their use in non-ruminant animals.
If polysorbate surfactants are combined with enzymes, water activity (Aw) becomes important. High levels of Aw, e.g., Aw>0.04, can significantly destabilize the shelf life of certain enzymes. Polysorbates are viscous, sticky, non-aqueous liquids that are not available in dry form. Before incorporation into enzyme formulations, polysorbates must be rendered into dry powders and the Aw of any formulation containing them should be below 0.04.
“Internal Probiotic Culture” or IPC, as used herein, is the totality of viable probiotic microorganisms present in the intestinal tract at any given time: i.e., the sum of probiotic microorganisms adhering to intestinal epithelial cells, mucous and mucin layers, ingested food and waste material. It is desirable to enhance the IPC of humans and mammals. IPC can be estimated from the total viable Lactobacillus and/or Bifidobacteria counts (colony forming units) present in fresh fecal matter.