Bacteria

Abstract

The present invention relates to bacteria and metabolites thereof that are capable of binding to A, B and/or O blood type antigens or which are acid and/or bile tolerant, their use in probiotic compositions and food products, and methods for their selection. The invention also relates to the use of said bacteria and metabolites for the prevention and/or treatment of gastrointestinal disorders.

1 - 15 . (canceled) 
     
     
         16 . A method of preventing or treating a gastrointestinal disorder comprising administering to a subject in need thereof a pharmaceutically effective amount of a bacteria or metabolite thereof, wherein the bacteria are characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or   e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay.   
     
     
         17 . The method of  claim 16 , wherein the method comprises administering to the subject a pharmaceutically effective amount of the bacteria. 
     
     
         18 . The method of  claim 17 , wherein the bacteria are characterized as having an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199. 
     
     
         19 . The method of  claim 17 , wherein the bacteria are characterized as having an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199. 
     
     
         20 . The method of  claim 17 , wherein the bacteria are characterized as having an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199. 
     
     
         21 . The method of  claim 17 , wherein the bacteria are characterized as having greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay. 
     
     
         22 . The method of  claim 17 , wherein the bacteria are characterized as having greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
     
     
         23 . The method of  claim 17 , wherein the method further comprises administering to the subject  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 or  Bifidobacterium lactis  Bi-07. 
     
     
         24 . The method of  claim 17 , wherein the bacteria comprise a mutant, variant or progeny of DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115 characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199,   d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or   e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay.   
     
     
         25 . The method of  claim 17  comprising administering to the subject a combination of two bacterial strains, wherein each of the bacterial strains is characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or 
 e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
 
     
     
         26 . The method of  claim 16 , wherein the gastrointestinal disorder comprises inflammatory bowel disease, irritable bowel syndrome, Chrone's disease, ulcerative colitis, constipation or diarrhea. 
     
     
         27 . A method of preventing or treating a gastrointestinal disorder comprising administering to a subject in need thereof a pharmaceutically effective amount of bacteria or a metabolite thereof, wherein the bacteria comprise DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115. 
     
     
         28 . A method of  claim 27 , wherein the method comprises administering to the subject a pharmaceutically effective amount of DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115. 
     
     
         29 . The method of  claim 28 , wherein the method further comprises administering to the subject  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 or  Bifidobacterium lactis  Bi-07. 
     
     
         30 . The method of  claim 29 , wherein the method comprises administering to the subject DSM 32109,  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 and  Bifidobacterium lactis  Bi-07. 
     
     
         31 . The method of  claim 27 , wherein the gastrointestinal disorder comprises inflammatory bowel disease, irritable bowel syndrome, Chrone's disease, ulcerative colitis, constipation or diarrhea. 
     
     
         32 . A probiotic composition, wherein:
 the probiotic composition comprises:
 bacteria or a metabolite thereof, and 
 a carrier; and 
   the bacteria are characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or 
 e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
   
     
     
         33 . The probiotic composition of  claim 32  comprising bacteria characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or 
 e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
 
     
     
         34 . The probiotic composition of  claim 33  further comprising  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 or  Bifidobacterium lactis  Bi-07. 
     
     
         35 . The probiotic composition of  claim 33  comprising a combination of two bacterial strains, wherein each of the bacterial strains is characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or 
 e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
 
     
     
         36 . The probiotic composition of  claim 33  comprising a mutant, variant or progeny of DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115 characterized as having:
 a) an average A antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 b) an average B antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 c) an average O antigen adhesion of greater than 100% when compared to  L. crispatus  LMG18199, 
 d) greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours as defined in an acid resistance assay, and/or 
 e) greater than 40% average bile tolerance represented as growth in 0.9% dehydrated fresh bile as a percentage of growth in MRS without bile as defined in a bile resistance assay. 
 
     
     
         37 . A probiotic composition, wherein:
 the probiotic composition comprises:
 bacteria or a metabolite thereof, and 
 a carrier; and 
   the bacteria comprise DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115.   
     
     
         38 . The probiotic composition of  claim 37  comprising DSM 32111, DSM 32108, DSM 32107, DSM 32098, DSM 32104, DSM 32112, DSM 32109, DSM 32105, DSM 32110, DSM 32103, DSM 32099, DSM 32106, DSM 32097, DSM 32114 or DSM 32115. 
     
     
         39 . The probiotic composition of  claim 38  further comprising  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 or  Bifidobacterium lactis  Bi-07. 
     
     
         40 . The probiotic composition of  claim 39  comprising DSM 32109,  Lactobacillus acidophilus  NCFM,  Bifidobacterium lactis  BL-04,  Lactobacillus paracasei  LPC37,  Bifidobacterium lactis  HNO19 and  Bifidobacterium lactis  Bi-07. 
     
     
         41 . A food product, dietary supplement, medical food or nutraceutical, wherein the food product, dietary supplement, medical food or nutraceutical comprises the probiotic composition of  claim 32 . 
     
     
         42 . The food product, dietary supplement, medical food or nutraceutical of  claim 41  comprising a nutritionally acceptable carrier, nutritionally acceptable diluent, nutritionally acceptable excipient, nutritionally acceptable adjuvant or nutritionally active ingredient in addition to the bacteria or metabolite. 
     
     
         43 . The food product of  claim 42  comprising a soft drink, fruit juice, a beverage comprising whey protein, health tea, cocoa drink, milk drink, lactic acid bacteria drink, yoghurt, cheese, ice cream, water ice, confectionery, biscuit, cake, snack food, balanced food, balanced drink, fruit filling, glaze, chocolate bakery filling, cheese cake flavored filling, fruit flavored cake filling, cake icing, doughnut icing, instant bakery filling cream, cookie filling, ready-to-use bakery filling, reduced-calorie filling, adult nutritional beverage, acidified soy/juice beverage, aseptic/retorted chocolate drink, bar mix, beverage powder, calcium-fortified milk or calcium-fortified coffee. 
     
     
         44 . The food product of  claim 42  comprising American cheese sauce, anti-caking agent for grated or shredded cheese, chip dip, cream cheese, dry-blended whip topping, fat-free sour cream, freeze/thaw dairy whipping cream, freeze/thaw stable whipped topping, low fat or light natural cheddar cheese, low fat Swiss style yoghurt, aerated frozen dessert, hard pack ice cream, low-fat ice cream, soft serve ice cream, barbecue sauce, cheese dip sauce, cottage cheese dressing, dry mix Alfredo sauce, mixed cheese sauce or dry mix tomato sauce. 
     
     
         45 . A food product, dietary supplement, medical food or nutraceutical wherein the food product, dietary supplement, medical food or nutraceutical comprises the probiotic composition of  claim 37 . 
     
     
         46 . A pharmaceutical composition, wherein the pharmaceutical composition comprises the probiotic composition of  claim 32 . 
     
     
         47 . The pharmaceutical composition of  claim 46  comprising a pharmaceutically acceptable carrier, pharmaceutically acceptable diluent, pharmaceutically acceptable excipient, pharmaceutically acceptable adjuvant or pharmaceutically active ingredient in addition to the bacteria or metabolite. 
     
     
         48 . A pharmaceutical composition, wherein the pharmaceutical composition comprises the probiotic composition of  claim 37 . 
     
     
         49 . A method of making the probiotic composition of  claim 32  comprising combining the bacteria or metabolite with the carrier. 
     
     
         50 . A method of making the probiotic composition of  claim 37  comprising combining the bacteria or metabolite with the carrier. 
     
     
         51 . A method of selecting bacteria or a metabolite thereof for treating or preventing a gastrointestinal disorder, wherein the method comprises:
 a) admixing test bacteria or a metabolite thereof with an A antigen, B antigen, O antigen or a combination thereof;   b) admixing  Lactobacillus reuteri  RC-14 control bacteria with the A antigen, B antigen, 0 antigen or combination thereof;   c) incubating said admixtures of step a) and b);   d) comparing the affinity of adhesion to said A antigen, B antigen, O antigen or combination thereof of the test bacteria or metabolite thereof and  Lactobacillus reuteri  RC-14; and   e) selecting bacteria (or a metabolite thereof) having a higher affinity of adhesion to said A antigen, B antigen, O antigen or combination thereof than  Lactobacillus reuteri  RC-14.   
     
     
         52 . A method of selecting bacteria or a metabolite thereof for treating or preventing a gastrointestinal disorder, wherein the method comprises:
 assaying one or both of the following:
 the average acid tolerance of the bacteria at a pH of 2.5 for 1.5 hours, and 
 the average bile tolerance of the bacteria in 0.9% bile; and 
   selecting bacteria (or a metabolite thereof) having:
 greater than −2.6 average acid tolerance represented as a log change when measured at a pH of 2.5 for 1.5 hours, and/or 
 greater than 40% average bile tolerance represented as growth in 0.9% bile as a percentage of growth in MRS without bile.