Probiotic bacterial molecules and their use in methods to treat/prevent infection by harmful bacteria and to provide nutritional health

Abstract

This invention provides isolated and characterized secreted molecules from probiotic bacteria for use in compositions and methods for the treatment and/or prevention of infection by harmful pathogenic bacteria. The isolated secreted molecules can also be used in nutritional or medical food products which provide probiotics to the gastrointestinal tract of a mammal.

1 . A composition for attenuation of virulence and/or treatment of an enteric bacterial infection in mammals, the composition comprising an effective amount of isolated secreted molecules from probiotic bacteria that attenuate the virulence of and/or treat enteric bacterial infection in mammals, wherein said probiotic bacteria is  Lactobacillus acidophilus  strain La-5, wherein the secreted molecules are peptides having from 2 to 10 amino acids and are not bacteriocins, and wherein the composition is concentrated. 
     
     
         2 . The composition of  claim 1 , wherein said composition is effective against  Salmonella  and/or  Escherichia coli  colonization. 
     
     
         3 . The composition of  claim 2 , wherein said  Salmonella  is  Salmonella enterica  and said  Escherichia coli  is EHEC O157:H7. 
     
     
         4 . The composition of  claim 1 , further combined within an edible food product, nutritional supplement, and/or ingestible liquid, one or more strains of whole probiotic bacteria and combinations thereof. 
     
     
         5 . The composition of  claim 1 , wherein said secreted molecules comprise an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         6 . The composition of  claim 1  further comprising a sugar source, and/or one or more antibiotics. 
     
     
         7 . The composition of  claim 6 , wherein the sugar source comprises glucose. 
     
     
         8 . The composition of  claim 1 , wherein said composition is concentrated by lyophilization or spray-drying. 
     
     
         9 . Isolated molecules from  Lactobacillus acidophilus  strain La-5, said molecules comprising an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof, wherein said molecules are not bacteriocins and are effective for treatment and/or attenuation of virulence of enteric bacterial infection in mammals in vivo, and wherein said molecules are concentrated. 
     
     
         10 . The molecules of  claim 9 , wherein said molecules are combined with one or more of an antibiotic, a sugar source, an edible food product, a nutritional supplement and ingestible liquid. 
     
     
         11 . A probiotic bacterial isolated proteinaceous fraction, said proteinaceous fraction comprising secreted molecules from Lactobacillus acidophilus strain La-5 in an amount effective in the treatment and/or attenuation of virulence of enteric bacterial infection in mammals in vivo, wherein said secreted molecules are proteinaceous having from 2 to 10 amino acids and are not bacteriocins and have one or more of the following characteristics:
 can withstand heating at up to about 90° C., freezing, thawing, lyophilization and/or spray drying and decrease virulence-related gene-expression, and wherein said proteinaceous fraction is concentrated.   
     
     
         12 . A food product, beverage product, health product, medicament or nutritional supplement that comprises one or more secreted molecules from a composition of  claim 1  in an amount effective for attenuation of virulence and/or treatment of an enteric bacterial infection in mammals and further optionally comprising one or more strains of whole probiotic bacteria. 
     
     
         13 . The food product, beverage product, health product, medicament or nutritional supplement of  claim 12 , wherein said molecules comprise an amino acid sequence selected from the group consisting of YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         14 . A method for attenuation of virulence of and/or therapeutically treating infections by  Escherichia coli  O157:H7 and/or  Salmonella  in a mammal, the method comprising administering to said mammal an effective amount of a composition of  claim 1  and optionally an antibiotic. 
     
     
         15 . The method of  claim 14 , wherein said secreted molecules comprise an amino acid sequence selected from the group consisting of YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         16 . A method for reducing the carriage by a food production animal of  Salmonella  strains that cause human salmonellosis, said method comprising administering an effective amount of the composition of  claim 1  to the food production animal prior to exposure to Salmonella strains that cause human salmonellosis. 
     
     
         17 . A method of reducing colonization by harmful bacteria in a mammal, the method comprising administration of an effective amount the composition of  claim 1  to said mammal. 
     
     
         18 . The method of  claim 17 , wherein said molecules inhibit the colonization of  E. coli  O157:H7 and/or  Salmonella.    
     
     
         19 . The method of  claim 18 , wherein said method further comprises the administration of a probiotic bacterium. 
     
     
         20 . The composition of  claim 5 , wherein said secreted molecules consist of an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         21 . The molecules of  claim 9 , said molecules consist of an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         22 . The food product, beverage product, health product, medicament or nutritional supplement of  claim 13 , wherein said molecules consist of an amino acid sequence selected from the group consisting of YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         23 . The method of  claim 15 , wherein said secreted molecules consist of an amino acid sequence selected from the group consisting of YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof 
     
     
         24 . A composition comprising whole  Lactobacillus acidophilus  strain La-5 bacteria for use in attenuation of virulence and/or treatment of an enteric bacterial infection in a mammal, wherein the bacteria secrete small peptides of 2 to 10 amino acids into the composition and the peptides attenuate virulence of and/or treat the enteric bacterial infection, wherein the peptides are not bacteriocins, and wherein the composition is concentrated. 
     
     
         25 . A composition comprising whole  Lactobacillus acidophilus  strain La-5 bacteria for use in reducing colonization of harmful bacteria in a mammal, wherein the bacteria secrete small peptides of 2 to 10 amino acids into the composition and the peptides reduce colonization of the harmful bacteria, wherein the peptides are not bacteriocins, and wherein the composition is concentrated. 
     
     
         26 . The composition of  claim 1 , wherein said secreted molecules comprise an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof and wherein the amino acid sequence comprises at least one modification selected from the group consisting of a deletion, a substitution, an insertion, glycosylation, unglycolsylation, complexation with organic or inorganic salts, PEGylation, and combinations thereof, wherein the modification does not substantially reduce the activity of the molecules for reduction and/or attenuation of virulence of bacterial infection. 
     
     
         27 . The proteinaceous fraction of  claim 11 , wherein said secreted molecules comprise an amino acid sequence selected from YPVEPF, YPPGGP, YPPG, NQPY and combinations thereof and wherein the amino acid sequence comprises at least one modification selected from the group consisting of a deletion, a substitution, an insertion, glycosylation, unglycolsylation, complexation with organic or inorganic salts, PEGylation, and combinations thereof, wherein the modification does not substantially reduce the activity of the molecules for reduction and/or attenuation of virulence of bacterial infection. 
     
     
         28 . The composition of  claim 1 , wherein said composition is cell-free. 
     
     
         29 . The composition of  claim 1 , wherein the molecules are purified. 
     
     
         30 . The proteinaceous fraction of  claim 11 , wherein the molecules are purified. 
     
     
         31 . The food product, beverage product, health product, medicament or nutritional supplement of  claim 12 , wherein the food product is a baked product. 
     
     
         32 . The food product, beverage product, health product, medicament or nutritional supplement of  claim 12 , being a health product, medicament, or nutritional supplement. 
     
     
         33 . A composition for attenuation of virulence and/or treatment of an enteric bacterial infection in mammals, the composition comprising an effective amount of isolated secreted molecules from probiotic bacteria that attenuate virulence of and/or treat enteric bacterial infection in mammals, wherein said probiotic bacteria is  Lactobacillus acidophilus  strain La-5, wherein the secreted molecules are proteinaceous having from 2 to 10 amino acids and are not bacteriocins, and wherein the composition is concentrated.