Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Intentional mixture of two or more micro-organisms – cells,...
Reexamination Certificate
1998-05-29
2003-02-25
Marx, Irene (Department: 1657)
Drug, bio-affecting and body treating compositions
Whole live micro-organism, cell, or virus containing
Intentional mixture of two or more micro-organisms, cells,...
C424S093450, C424S093510, C424S093480
Reexamination Certificate
active
06524574
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates to a mixture of probiotics to be fed to monogastric animals and, more specifically, a mixture of facultative anaerobic probiotic organisms affecting and controlling or inhibiting the colonization of deleterious bacteria in the intestines of monogastric animals and humans.
2. Background of the Prior Art
Probiotics are defined as microbes that are fed to animals to improve the microbial populations in the intestines of animals or humans. Most prior art probiotics are lactic acid-producing bacteria. The probiotics of the present invention include both bacteria and yeasts.
Probiotics have been fed to animals to reduce or replace the potentially pathogenic intestinal bacteria with nonpathogenic species.
Spring, March 1997; (Animal Talk), opines that the main regulatory mechanisms used by natural gut inhibitants as Lactobacilli and Enterococci is to keep pathogenic bacteria from colonizing the digestive tract. Mechanisms discussed include competition for nutrients, growth factors, intestinal receptor sites, and stimulation of epithelial cell turnover. Creation of a restrictive environment includes lower pH, VFA and lactic acid production or induction of an immunologic process or antimicrobial substances.
Presser, et al, 1997 (Appl & Environ Microbio Vol. 63 No. 6:2355-2360), observed the negative effects on the growth rate of
E. coli
as a function of pH and lactic acid concentration. The findings demonstrated
E. coli
growth rate was linearly related to the hydrogen ion concentration. In the range 0 to 100 mM lactic acid, growth rate was also linearly related to the undissociated lactic acid concentration.
Hudault et al., 1997 (Appl & Environ Microbio Vol. 63 No. 6:513-518) demonstrated an antagonistic activity exerted in vitro and in vivo by
Lactobacillus casei
(strain GG) against
Salmonella typhimurium
C5 infection. The mechanism of this antagonist activity was reported to be dependent upon an acidic environment due to lactic acid itself or alternatively to a substance that is active at low pH.
Kimura, et al., 1997 (Applied and Environmental Microbiology Vol.63, No. 9:3394-3398) found that human test subjects harbored a unique collection of Bifidobacteria and Lactobacilli species. Their results emphasized the complexity of the relationship between the intestinal microflora and the host where it appears that the host's predominant strains do not illicit as large of a IgG titer response as do nondominant intestinal species.
Perdigon, et al., 1991 (J Dairy Research Vol. 58:485-496) showed an immunoadjuvant activity of oral
Lactobacillus casei
by playing an important role in the prevention of enteric infections by increasing IgA secretion into the intestinal lumen thus providing a defense for the mucosal surface against enteric (i.e., Salmonella and
E. coli
) challenges.
Bernet, et al., 1994 (Gut 35:483-489) demonstrated
Lactobacillus acidoiphilis
LA1 binds to cultured human intestinal cell walls and inhibits cell attachment and cell invasion by enterovirulent bacteria.
Fujiwara, et al., 1997 (Appl & Environ Microbio Vol. 63 No. 6:506-512) showed that the binding inhibitor produced by
B. longum
and other Bifidobacterium species was estimated to contribute to their normal anti-infectious activities by preventing the binding of pathogenic strains of
E. coli
to their common bacterium intestinal binding structures.
Firon, et al., 1983 (Carbohydrate Res. 120:235-249) commented on carbohydrate specificity of the surface lectins of Escherichia, Kleibsiella, and Salmonella organisms.
Adlerberth, et al., 1996 (Appl & Environ Microbio Vol. 63 No. 6:2244-2251) found mannose-specific adhesions in a variety of gram-negative bacteria including members of the family Enterobacteriaceae such as Escherichia, Shigella, Enterobacter, Kleibsiella, and Salmonella and in Pseudomonas and Vibrio.
Adlerberth, et al., also found that Saccharomyces species of yeast contain mannose- containing polysaccharides in their cell walls which
E. coli
and other intestinal bacteria adhere to in a mannose-specific manner.
Jonvel, 1993 (Feed Mix Vol. 1, No. 4) citing Gedek, stated there are three possible explanations of Saccharomyces yeasts' mode of action: 1) fixation of
E. coli
on the yeast cell wall surface if
E. coli
has fimbria (i.e., the fimbria have an affinity to mannose); 2) fixation of enterotoxin on the yeast cell wall surface and are destroyed; and 3)
E. coli
destruction by lethal yeast toxin.
Abe, et al. (1995. J Dairy Science 78:2838-2846) showed that probiotics fed to newborn calves and piglets decreased frequency of diarrhea, and stimulated body weight gains and feed conversion in those animals over control animals.
Saavedra, et al., 1994 (Lancet 334:1046-1049) showed that feeding of a Bifidobacteria and Enterococci species to hospitalized human infants for prevention of diarrhea and the shedding of rotavirus.
Ozawa, et al. 1983 (Applied and Environmental Microbiology 45: 151) reported that the administration of an Enterococcus species to calves and piglets promoted colonization of beneficial bacteria and decreased the occurrence of detrimental bacteria, such as Salmonella, in the intestine.
Surawiciz, et al., 1989 (Gastroenterology Vol. 96, No. 4: 961-968) showed that
Saccharomyces boulardii
significantly reduced the incidence of antibiotic-associated diarrhea in hospitalized patients. S. boulardii has antagonistic activity against a variety of bacterial pathogens. In rats and humans it increases the disaccharidase activity in intestinal mucosa which, in turn, may improve carbohydrate absorption within the host.
From the Bergey's Manual of Systematic Bacteriology 1984. it is observed that within the genera Campylobacter, Pseudomonas and most Vibrio and Clostridia species do not utilize lactose as a carbon source for growth. Also, Enterobacteriaceae, in general, are poor or nonutilizers of lactose. Whereas, the Enterococcus species used in the invention readily utilize lactose.
The importance of mannose-sensitive adhesions of gram-negative intestinal bacteria for intestinal colonization of these bacteria was investigated, as well as the presence of a mannose-specific adhesion in a gram-positive bacterial species, i.e., Lactobacilli and Enterococci, which belong to the indigenous intestinal microflora. Further, investigation was made into the ability of gram-positive, non-pathogenic Enterococci and Lactobacilli to associate with animals' intestines and mannose containing polysaccharides (mannose) found as a major cell wall component in species of Saccharomyces, and, in particular, its use in conjunction with the lactic acid-producing metabolism of Enterococci and/or Lactobacilli to rid and/or prophylatically protect the intestines of monogastric animals and humans of potentially pathogenic bacteria.
SUMMARY OF THE INVENTION
The invention consists of a defined combination of facultative anaerobic probiotic organisms effective in controlling or inhibiting the colonization of certain deleterious bacteria in the intestines of monogastric animals and humans. In a preferred embodiment of the invention, nutrients are added to the mixture of probiotics to enhance efficacy in some situations.
The invention is a unique probiotic mixture that combines two viable lactic acid-producing Enterococcus strains and two viable Saccharomyces yeast strains that are preferably added to an active carrier that includes nutrients to assist in the growth and/or activity of the probiotics. The mixture is fed to monogastric animals to control the microflora population of the intestinal tract and to maintain a proper balance of naturally occurring beneficial microflora while competing with and helping to exclude deleterious strains of microflora such as bacterial pathogens, thus aiding the ability of the animal to maintain a normal, healthy intestinal environment and, in turn, utilize feeds better.
Ingesting this probiotic mixture by monogastric animals and humans will help the body protect itself from colonization o
Brown Patrick K.
Spangler David A.
Witzig Thomas E.
Afremova Vera
Agri-King, Inc.
Clark H.
Laurenzo Brian J.
Marx Irene
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