Lactic acid producing bacteria for use as probiotic organisms in the human vagina

Abstract

The present invention discloses novel isolated strains of lactic acid producing bacteria of the genera Lactobacillus and Pediococcus , and a method for isolation of such bacterial strains, having the ability to colonise and become established in a human vagina, even during menstrual discharge. Furthermore, a composition comprising said bacterial strains and a sanitary article comprising said bacterial strains, such as a tampon, for prophylaxis and/or treatment of infections of the urogenital tract, are described. The present invention also describes a method for prophylaxis and/or treatment of infections of the urogenital tract, wherein at least one of said bacterial strains are administered vaginally.

1 . A method for isolation of a bacterial strain of the genus  Lactobacillus  or  Pediococcus  having the ability to colonise and become established in a human vagina, even during menstrual discharge, comprising the following steps: 
 a) a bacterial sample is collected from the vaginal tract of a woman with a normal, healthy vaginal flora of microorganisms,    b) lactic acid producing bacteria are selected from the vaginal sample of step a),    c) the lactic acid producing bacteria of step b) are pure cultured in vitro in a suitable nutrient medium providing at least one isolated bacterial strain,    d) at least one pure cultured bacterial strain of step c) and/or a combination of at least two bacterial strains of step c) is evaluated as to its ability to colonise and become established in the vagina upon vaginal administration of the bacterial strain to a woman during her menstrual discharge, said woman displaying a disturbed vaginal flora of microorganisms, wherein said bacterial strain or strains is/are considered established if the bacterial strain or strains is/are still present in the vagina after at least two menstrual cycles from the time of administration, and    e) at least one bacterial strain of step d) that displays said ability to colonise and become established in the vagina is selected.    
     
     
         2 . The method according to  claim 1 , comprising a further step of at least one pure cultured bacterial strain of step c) and/or a combination of at least two bacterial strains of step c) is lyophilised.  
     
     
         3 . The method according to  claim 2 , comprising a further step of at least one lyophilised, pure cultured bacterial strain of step c) and/or a combination of at least two lyophilised bacterial strains of step c), is evaluated and selected as to stability of bacterial viability upon lyophilisation and viability of lyophilised bacteria over long periods of storage, wherein said bacterial viability is considered stable if at least 40% of the bacteria have a preserved viability upon lyophilisation and storage of the lyophilised bacteria at −18° C. for at least 12 months.  
     
     
         4 . The method according to  claim 2 , comprising a further step of at least one lyophilised, pure cultured bacterial strain of step c) and/or a combination of at least two lyophilised bacterial strains of step c), is evaluated and selected as to genetic stability upon repeated cultivation.  
     
     
         5 . The method according to  claim 3 , comprising a further step of at least one lyophilised, pure cultured bacterial strain of step c) and/or a combination of at least two lyophilised bacterial strains of step c), is evaluated and selected as to genetic stability upon repeated cultivation.  
     
     
         6 . A method for prophylaxis and/or treatment of infections of the urogenital tract, wherein at least one of the bacterial strains according to  claim 1  is vaginally administered in a therapeutically effective amount to prevent, alleviate the effects of, and/or treat a microbial infection.  
     
     
         7 . The method according to  claim 5 , said method is for prophylaxis and/or treatment of bacterial vaginosis or any other bacterial disorder in the vagina.  
     
     
         8 . An isolated bacterial strain of the genus  Lactobacillus  or  Pediococcus  obtained by the method of  claim 1 , said  Lactobacillus  or  Pediococcus  is selected from the group consisting of the strain of  Lactobacillus gasseri , LN 40, deposited under number LMG P-20560, the strain of  Lactobacillus  casei subsp rhamnosus, LN 113, deposited under number LMG P-20562, the strain of  Lactobacillus fermentum , LN 99, deposited under number LMG P-20561, the strain of  Lactobacillus crispatus , LN 01, deposited under number LMG P-20558, and the strain of  Pediococcus acidilactici , LN 23, deposited under number LMG P-20559, and having the ability to colonise and become established in a human vagina upon vaginal administration, even during menstrual discharge, said strains were deposited at Belgian Coordinated Collections of Microorganisms on 14 June 2001.