Means and Methods for Preventing and/or Treating Caries

Abstract

The present invention relates to a microorganism belonging to the group of lactic acid bacteria capable of specifically binding to Streptococcus mutans , wherein the specific binding is (i) resistant to heat treatment; (ii) resistant to protease treatment; (iii) calcium-dependent; (iv) formed within a pH range between 4.5 and 8.5; and/or (v) formed in the presence of saliva. The present invention is also relates to an analog or fragment of the microorganism which is thermally inactivated or lyophilized, wherein the analog or fragment retains the capability of specifically binding to Streptococcus mutans . The invention further encompasses compositions and additives for food, feed or drinks comprising the microorganism or an analog or fragment thereof. Also provided is uses of the microorganism or an analog or fragment thereof for the preparation of an anticariogenic or pharmaceutical composition or anticariogenic food or feedstuff as well as methods for producing the compositions or food or feedstuff.

1 . An isolated microorganism belonging to the group of lactic acid bacteria, wherein said isolated microorganism binds to  Streptococcus mutans  in the presence of saliva or at a pH range between 4.5 and 8.5, and wherein the binding between said isolated microorganism and  Streptococcus mutans  is calcium-dependent, resistant to heat treatment at more than 95° C. for at least 20 minutes, or resistant to protease treatment. 
     
     
         2 . The isolated microorganism of  claim 1 , wherein said microorganism binds to  Streptococcus mutans  in the presence of saliva and at a pH range between 4.5 and 8.5, and wherein the binding between said microorganism and  Streptococcus mutans  is calcium-dependent, resistant to heat treatment at more than 95° C. for at least 20 minutes, and resistant to protease treatment. 
     
     
         3 . The isolated microorganism of  claim 1 , wherein said microorganism is of the genus of  Lactobacillus.    
     
     
         4 . The isolated microorganism of  claim 1 , wherein said microorganism is  Lactobacillus paracasei  or  Lactobacillus rhamnosus.    
     
     
         5 . The isolated microorganism of  claim 1 , wherein said microorganism is capable of binding to  Streptococcus mutans  serotype c (DSMZ 20523) and/or serotype e (NCTC 10923) and/or serotype f (NCTC 11060). 
     
     
         6 . The isolated microorganism of  claim 1 , wherein said microorganism is live, dead, thermally inactivated, lyophilized, spray-dried, or dried. 
     
     
         7 . An analog or fragment of the isolated microorganism of  claim 1  which is thermally inactivated or lyophilized, wherein said analog or fragment retains the capability of specifically binding to  Streptococcus mutans  as said isolated microorganism. 
     
     
         8 . A composition comprising the isolated microorganism of  claim 1 , or an analog or fragment thereof, in an amount:
 (a) between 10 2  to 10 12  cells per mg in a solid form of the composition,   (b) between 10 3  to 10 8  cells per mg in a solid form of the composition, or   (c) between 10 2  to 10 13  cells per ml,   wherein said composition optionally comprises no more than 1% (w/w) lactose or more than 6% (w/w) lactose.   
     
     
         9 . The composition of  claim 8 , wherein the composition is solid, liquid or gaseous form. 
     
     
         10 . The composition of  claim 8 , wherein the composition is in form of powder, tablet, solution, aerosol, granule, pill, suspension, emulsion, capsule, syrup, liquid, elixir, extract, tincture or fluid extract. 
     
     
         11 . The composition of  claim 8 , wherein the composition is:
 (a) a cosmetic composition further comprising a cosmetically acceptable carrier or excipient,   (b) a pharmaceutical composition further comprising a pharmaceutical acceptable carrier or excipient, or   (c) a food or feed composition further comprising an orally acceptable carrier or excipient.   
     
     
         12 . The composition of  claim 8 , wherein the composition has anticariogenic activity. 
     
     
         13 . The composition of  claim 8 , wherein the composition is in form of a dentifrice, chewing gum, lozenge, mouth wash, mouth rinse or dental floss. 
     
     
         14 . An isolated microorganism belonging to the group of lactic acid bacteria isolated from an assay comprising:
 (a) growing a microorganism belonging to the group of lactic acid bacteria to stationary phase;   (b) mixing said microorganism with  Streptococcus mutans  which has been grown to stationary phase;   (c) incubating the mixture of said microorganism and  Streptococcus mutans  obtained in step (b) under conditions allowing formation of aggregates of said microorganism and  Streptococcus mutans ; and   (d) detecting aggregates by occurrence of a pellet,   wherein, prior to mixing with  Streptococcus mutans , the microorganism is:   (i) heated at more than 95° C. for at least 20 minutes,   (ii) lyophilized at room temperature under vacuum for about two hours,   (iii) treated with a protease, or   (iv) resuspended in a solution comprising about 2 mM of calcium.   
     
     
         15 . The isolated microorganism of  claim 14 , wherein the formation of aggregates of the microorganism and  Streptococcus mutans  occurs in the presence of saliva or at a pH range between 4.5 and 8.5. 
     
     
         16 . The isolated microorganism of  claim 14 , wherein said microorganism is of the genus of  Lactobacillus.    
     
     
         17 . The isolated microorganism of  claim 14 , wherein said microorganism is  Lactobacillus paracasei  or  Lactobacillus rhamnosus.    
     
     
         18 . The isolated microorganism of  claim 14 , wherein said microorganism is live, dead, thermally inactivated, lyophilized, spray-dried, or dried. 
     
     
         19 . An analog or fragment of the isolated microorganism of  claim 14  which is thermally inactivated or lyophilized, wherein said analog or fragment retains the capability of specifically binding to  Streptococcus mutans  as said isolated microorganism. 
     
     
         20 . A composition comprising the isolated microorganism of  claim 13 , or an analog or fragment thereof, in an amount:
 (a) between 10 2  to 10 12  cells per mg in a solid form of the composition,   (b) between 10 3  to 10 8  cells per mg in a solid form of the composition, or   (c) between 10 2  to 10 13  cells per ml,   wherein said composition optionally comprises no more than 1% (w/w) lactose or more than 6% (w/w) lactose.   
     
     
         21 . The composition of  claim 20 , wherein the composition is solid, liquid or gaseous form. 
     
     
         22 . The composition of  claim 20 , wherein the composition is in form of powder, tablet, solution, aerosol, granule, pill, suspension, emulsion, capsule, syrup, liquid, elixir, extract, tincture or fluid extract. 
     
     
         23 . The composition of  claim 20 , wherein the composition is:
 (a) a cosmetic composition further comprising a cosmetically acceptable carrier or excipient,   (b) a pharmaceutical composition further comprising a pharmaceutical acceptable carrier or excipient, or   (c) a food or feed composition further comprising an orally acceptable carrier or excipient.   
     
     
         24 . The composition of  claim 20 , wherein the composition has anticariogenic activity. 
     
     
         25 . The composition of  claim 20 , wherein the composition is in form of a dentifrice, chewing gum, lozenge, mouth wash, mouth rinse or dental floss. 
     
     
         26 . A method for the production of an anticariogenic composition comprising obtaining the isolated microorganism of  claim 1 , or an analog or fragment thereof, and formulating said microorganism, or the analog or fragment thereof, with a cosmetically, pharmaceutical or orally acceptable carrier or excipient. 
     
     
         27 . A method for the production of an anticariogenic foodstuff or feedstuff comprising obtaining the isolated microorganism of  claim 1 , or an analog or fragment thereof, and formulating said microorganism, or the analog or fragment thereof, to a foodstuff or feedstuff. 
     
     
         28 . A method for the production of an anticariogenic composition comprising obtaining the isolated microorganism of  claim 13 , or an analog or fragment thereof, and formulating said microorganism, or the analog or fragment thereof, with a cosmetically, pharmaceutical or orally acceptable carrier or excipient. 
     
     
         29 . A method for the production of an anticariogenic foodstuff or feedstuff comprising obtaining the isolated microorganism of  claim 13 , or an analog or fragment thereof, and formulating said microorganism, or the analog or fragment thereof, to a foodstuff or feedstuff.