Probiotic strains, a process for the selection of them, compositions thereof, and their use

Abstract

The present invention relates to a novel process for the selection of new probiotic strains which comprises the following steps: a) selecting for non-pathogenic strains which are capable of surviving in breast milk and/or amniotic fluid, and b) selecting for non-pathogenic strains which are able to be transferred to breast milk and/or amniotic fluid after oral intake in healthy individuals without colonizing other internal organs except mucousas. The invention also provides new Lactobacillus strains, which are: CECT5711 ( Lactobacillus coryniformis ), CECT5713 ( Lactobacillus salivarius subsp. salivarius), CECT5714: ( Lactobacillus gasseri , formerly L. acidophilus ), CETC5715: ( Lactobacillus gasseri ), and CECT5716: ( Lactobacillus fermentum ); and refers to their use for the prophylaxis or treatment against digestive, infective, neuro-degenerative and immune related diseases such as allergies or inflammatory diseases.

1 . A method for the selection of probiotic microbial strains, comprising the following steps: 
 a. selecting for non-pathogenic strains which are capable of surviving in breast milk and/or amniotic fluid, and    b. selecting for non-pathogenic strains which are able to be transferred to breast milk and/or amniotic fluid after oral intake in healthy individuals without colonizing other internal organs except mucousas.    
     
     
         2 . A method according to  claim 1 , wherein both breast milk and amniotic fluid are from human sources.  
     
     
         3 . A method as claimed in  claim 2 , wherein the probiotic tested strains are any lactic acid bacteria selected from the genera: Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Streptococcus and Bifidobacterium.  
     
     
         4 . A method according to claims  3 , wherein the probiotic strains tested had previously been obtained from: breast milk, feces of breastfed babies or amniotic fluid.  
     
     
         5 . A method according to  claim 4 , wherein the strains tested had been obtained from human samples.  
     
     
         6 . A bacterial strain selected by a method according to any of claims  1  to 5  
     
     
         7 . A strain of bacteria deposited in the CECT under Accession N o  5711 ( Lactobacillus coryniformis ) or a variant thereof.  
     
     
         8 . A strain of bacteria deposited in the CECT under Accession N o  5713 ( Lactobacillus salivarius  subsp. salivarius) or a variant thereof.  
     
     
         9 . A strain of bacteria deposited in the CECT under Accession N o  5714 ( Lactobacillus gasseri,  formerly  L. acidophilus ) or a variant thereof.  
     
     
         10 . A strain of bacteria deposited in the CECT under Accession N o  5715 ( Lactobacillus gasseri ) or a variant thereof.  
     
     
         11 . A strain of bacteria deposited in the CECT under Accession Nu o  5716 ( Lactobacillus fermentum ) or a variant thereof.  
     
     
         12 . A biologically pure culture of a strain according to any of claims  6 - 11 .  
     
     
         13 . Use of mammal milk and of mammal amniotic fluid as a source to obtain non-pathogenic probiotic bacteria, especially probiotic bacteria selected from the genera Lactobacillus, Lactococcus, Enterococcus, Streptococcus and Bifidobacterium  
     
     
         14 . Use of mammal milk and of mammal amniotic fluid according to  claim 13  where the mammal milk and the mammal amniotic fluid are human.  
     
     
         15 . Use of mammal milk and of mammal amniotic fluid according to  claim 14  as a source to obtain the microbial strains defined in any of the  claims 6  to  11 .  
     
     
         16 . A composition comprising at least one of the bacterial strains defined in  claims 6  to  11 , where the composition comprises preferably from 2 to 6 strains, more preferably from 2 to 4 strains, most preferably from 2 to 3 strains, and where each of the strains is present in the composition in a proportion from 0.1% to 99.9%, preferably from 1% to 99%, more preferably from 10% to 90%.  
     
     
         17 . A composition comprising at least one of the bacterial strains defined in  claims 6  to  11  together with another strain or mixture of strains where the mixture comprises preferably from 2 to 6 strains, more preferably from 2 to 4 strains, most preferably from 2 to 3 strains and where each of the strains is present in the composition in a proportion from 0.1% to 99.9%, preferably from 1% to 99%, more preferably from 10% to 90%.  
     
     
         18 . A composition comprising a strain as claimed in any one of  claims 6  to  11  or a composition as defined in any of the  claims 16  to  17  in a lyophilized form.  
     
     
         19 . A composition comprising a strain as claimed in any one of  claims 6  to  11  or a composition as defined in any of the  claims 16  to  17  in a frost form.  
     
     
         20 . A composition comprising a strain as claimed in any one of  claims 6  to  11  or a composition as defined in any of the  claims 16  to  17  in an inactivated form or dead.  
     
     
         21 . A composition obtainable from the supernatant of a culture of a bacterial strain according to any of the  claims 6  to  11 , or from a composition according to any of the  claims 16  to  17 .  
     
     
         22 . A composition obtainable by extraction of a culture of any of the bacterial strains according to  claims 6  to  11 , or by extraction of a composition according to  claims 16  to  17 .  
     
     
         23 . A product obtainable from the metabolic activity of any of the strains specified in  claims 6  to  11 , from a culture according to  claim 12  or from a composition according to  claims 16  to  17 , wherein the product is preferably an enzyme.  
     
     
         24 . Food product comprising support material and at least one strain according to any of the  claims 6  to  11 , a culture, a composition or a product according to claims  12  and  16  to  24 .  
     
     
         25 . A product according to  claim 24  wherein the support material is a food composition selected from milk, yoghourt, curd, cheese, fermented milks, milk based fermented products, meat based fermented products, fermented cereals based products, milk based powders, cereal based powders, infant formulae, clinical nutrition formula, ice-creams, juices, flours, bread, cakes, sugar, candies or chewing-gums.  
     
     
         26 . A product according to  claim 24 , wherein the microbial strain according to  claims 6  to  11  is contained in the support material in an amount from about 10 5  cfu/g to about 10 12  cfu/g support material, preferably from about 10 6  cfu/g to about 10 11  cfu/g support material, more preferably from about 10 6  cfu/g to about 10 10  cfu/g support material.  
     
     
         27 . A pharmaceutical composition comprising at least one strain according to any of the  claims 6  to  11 , a culture, a composition or a product according to claims  12  and  16  to  26  and pharmaceutically acceptable excipients.  
     
     
         28 . A pharmaceutical composition according to  claim 27  wherein the microbial strains are contained in an amount from 10 5  cfu/g to 10 14  cfu/g support material, preferably from 10 6  cfu/g to 10 13  cfu/g support material, more preferably from 10 7  cfu/g to 10 12  cfu/g support material.  
     
     
         29 . A strain of bacteria as claimed in anyone of  claims 6  to  11 , a culture, a composition or product according to claims  12  and  16  to  26  for therapeutic or prophylactic treatment.  
     
     
         30 . A composition according to  claim 29  for topic, oral, ocular, nasal, enteral, urogenital, vaginal or rectal administration  
     
     
         31 . A composition according to  claim 29  designed to be administered to pregnant woman for the therapeutic or prophylactic treatment of their foetus.  
     
     
         32 . A composition according to  claim 29  designed to be administered to lactating woman for the therapeutic or prophylactic treatment of their breastfed babies.  
     
     
         33 . Use of a strain according to any of the  claims 6  to  11 , a culture, a composition or a product according to claims  12  and  16  to  27  in the manufacture of a product for the therapeutic or prophylactic treatment of human and animal diseases.  
     
     
         34 . Use according to  claim 33  for the treatment and/or prophylaxis of chronic or acute infection, or of undesirable microbial colonization, wherein the infection or colonization is caused by parasites, bacteria, yeasts, fungi or viruses, of a mucosal surface in a subject or animal in need thereof, wherein the mucosal surface is selected from but not restricted to the group consisting of oral, nasopharyngeal, respiratory, gastric, intestinal, urogenital and glandular.  
     
     
         35 . Use according to  claim 33  for the treatment and/or prophylaxis of temporally depressed immune levels in individuals subjected to physiological stress.  
     
     
         36 . Use according to  claim 33  for the improvement of the immune gut barrier in a subject or animal in need thereof; for the treatment and/or prophylaxis of down-regulating hypersensitivity reactions to food and metabolic intolerance such as: lactose intolerance; of constipation and other gastro-intestinal disorders; of inflammatory or auto-immune disorders such as: 113D, ulcerative colitis, arthritis, atherosclerosis, multiple sclerosis, psoriasis or sarcoidosis; and of tumor growth, metastasis and cancer in a subject or animal in need thereof.  
     
     
         37 . Use according to  claim 33  for the treatment and/or prophylaxis of allergic disorders and asthma in a subject or animal in need thereof.  
     
     
         38 . Use according to  claim 33  for the treatment and/or prophylaxis of neuro-degenerative diseases in an individual or animal in need thereof, selected from, but not restricted to, the group consisting of Parkinson, stroke, Alzheimer, Huntington and dementia.