Bacterial strains isolated from pigs

Patent metadata

Field	Value
Publication number	US-2017173089-A1
Application number	US-201615359144-A
Country	US
Kind code	A1
Filing date	Nov 22, 2016
Priority date	Jul 14, 2011
Publication date	Jun 22, 2017
Grant date	—

Abstract

A first aspect of the invention relates to a porcine lactic acid bacterial strain, wherein said bacterial strain is characterised by one or more of the following characteristics: (i) the ability to exhibit antimicrobial activity against E. coli ; (ii) the ability to exhibit antimicrobial activity against S. enteritidis ; (iii) the ability to suppress inflammation in IPEC cells induced by 12-0-tetradecaboylphorbol-13-acetate (PMA); (iv) the ability to block the attachment or invasion of IPEC cells by S. enteritidis ; (v) the ability to block the attachment or invasion of IPEC cells by E. coli ; (vi) the absence of antibiotic resistance to one or more antibiotics selected from the following: ampicillin; cefotaxime; chloramphenicol; erythromycin; gentamicin; tetracycline; vancomycin; metronizadole; nalidixic acid; and kanamycin; and (vii) the ability to exhibit heat stability when subjected to three cycles of heating, each cycle comprising heating at a temperature of 70° C. for a period of 15 minutes. Further aspects of the invention relate to compositions comprising said bacterial strains, and therapeutic uses of said bacterial strains.

First claim

1 . A pharmaceutical composition comprising:
at least one porcine lactic acid bacterial strain; wherein the at least one porcine lactic acid bacterial strain is in an amount sufficient to treat or prevent an intestinal disorder in a subject; wherein said pharmaceutical composition exhibits heat stability when subjected to three cycles of heating, each cycle comprising heating at a temperature of 70° C. for a period of 15 minutes, wherein said heat stability is determined by an ability of the at least one porcine lactic acid bacterial strain to block adherence of a pathogen to an intestinal pig epithelial cell (IPEC) in vitro; and
wherein said porcine lactic acid bacterial strain is lyophilized and/or said pharmaceutical composition further comprises a pharmaceutically acceptable excipient, diluent, or carrier.

2 . The pharmaceutical composition of claim 1 , wherein the at least one porcine lactic acid bacterial strain exhibits at least one characteristic selected from a group consisting of:
(i) antimicrobial activity against E. coli; (ii) antimicrobial activity against S. enteritidis; (iii) suppression of inflammation in IPEC cells induced by 12-O-tetradecaboylphorbol-13-acetate (PMA); (iv) an ability to block the attachment or invasion of IPEC cells by S. enteritidis; (v) an ability to block the attachment or invasion of IPEC cells by E. coli ; and (vi) absence of antibiotic resistance to an antibiotic selected from the group consisting of: ampicillin; cefotaxime; chloramphenicol; erythromycin; gentamicin; tetracycline; vancomycin; metronizadole; nalidixic acid; and kanamycin.

3 . The pharmaceutical composition of claim 2 , wherein the at least one porcine lactic acid bacterial strain exhibits any two characteristics selected from the group of claim 2 .

4 . The pharmaceutical composition of claim 2 , wherein the at least one porcine lactic acid bacterial strain exhibits any three characteristics selected from the group of claim 2 .

5 . The pharmaceutical composition of claim 2 , wherein the at least one porcine lactic acid bacterial strain exhibits any four characteristics selected from the group of claim 2 .

6 . The pharmaceutical composition of claim 2 , wherein the at least one porcine lactic acid bacterial strain exhibits any five characteristics selected from the group of claim 2 .

7 . The pharmaceutical composition of claim 2 , wherein the at least one porcine lactic acid bacterial strain exhibits all six characteristics in the group of characteristics in claim 2 .

8 . The pharmaceutical composition of claim 1 , wherein the at least one porcine lactic acid bacterial strain is selected from the group consisting of: L. johnsonii, L. reuteri, L. plantarum, L. gasseri, L. pentosus, L. acidophilus, L. vaginalis, L. mucosae , and any combination thereof.

9 . The pharmaceutical composition of claim 8 , comprising a combination of porcine lactic acid bacteria strains; wherein the combination of porcine lactic acid bacterial strains comprises L. johnsonii , and L. reuteri.

10 .- 17 . (canceled)

18 . A method of treating an intestinal disorder in a subject, said method comprising administering to the subject a pharmaceutical composition comprising at least one porcine lactic acid bacterial strain; wherein the at least one porcine lactic acid bacterial strain is in an amount sufficient to treat or prevent an intestinal disorder in a subject wherein said pharmaceutical composition exhibits heat stability when subjected to three cycles of heating, each cycle comprising heating at a temperature of 70° C. for a period of 15 minutes, wherein said heat stability is determined by an ability of the at least one porcine lactic acid bacterial strain to block adherence of a pathogen to an IPEC in vitro.

19 . (canceled)

20 . A method of improving intestinal microbiota in a subject in need thereof, said method comprising administering to the subject a pharmaceutical composition comprising: at least one porcine lactic acid bacterial strain in an amount sufficient to improve intestinal microbiota in said subject wherein said pharmaceutical composition exhibits heat stability when subjected to three cycles of heating, each cycle comprising heating at a temperature of 70° C. for a period of 15 minutes, wherein said heat stability is determined by an ability of the at least one porcine lactic acid bacterial strain to block adherence of a pathogen to an IPEC in vitro.

21 . (canceled)

22 . A feedstuff comprising the pharmaceutical composition of claim 1 .

23 . A food product comprising the pharmaceutical composition of claim 1 .

24 . A dietary supplement comprising the pharmaceutical composition of claim 1 .

25 . A food additive comprising the pharmaceutical composition of claim 1 .

26 . A process for producing a probiotic composition, said process comprising:
(a) culturing at least one porcine lactic acid bacterial strain in a culture medium and recovering said at least one porcine lactic acid bacterial strain from the culture medium; wherein said at least one porcine lactic acid bacterial strain comprises a 16S rRNA gene sequence with at least 93% homology to a 16s rRNA gene sequence of any one of SEQ ID NOs 1-87; and (b) mixing the culture of (a) with an excipient, diluent or carrier.

27 . A method of preparing at least one porcine lactic acid bacterial strain, said method comprising the steps of:
(i) obtaining faeces from an organically reared pig; (ii) freezing the faeces and dispersing in a suitable diluent; (iii) applying the dispersed faeces obtained in step (ii) to a suitable agar, optionally in the presence of supplemental pig colostrum carbohydrates, and incubating under an anaerobic conditions; (iv) selecting off distinct colonies of bacteria formed during step (iii) and seeding into a suitable broth, optionally in the presence of supplemental pig colostrum carbohydrates; (v) incubating the seeded colonies obtained in step (iv); and (vi) obtaining an aliquot of the incubated broth, said aliquot comprising at least one porcine lactic acid bacterial strain; wherein the at least one porcine lactic acid bacterial strain comprises a 16S rRNA gene sequence with at least 93% homology to a 16s rRNA gene sequence of any one of SEQ ID NOs 1-87.

28 .- 29 . (canceled)

30 . One or more porcine lactic acid bacterial strains obtained by, or obtainable by, the process of claim 27 .

31 . A pharmaceutical composition comprising:
a porcine lactic acid bacterial strain comprising a 16S rRNA gene sequence with at least 93% homology to a 16s rRNA gene sequence of any one of SEQ ID NOs 1-87; wherein said porcine lactic acid bacterial strain is lyophilized and/or said pharmaceutical composition further comprises a pharmaceutically acceptable excipient, diluent, or carrier; and
wherein said pharmaceutical composition is in unit dose form.

32 . The pharmaceutical composition of claim 1 , comprising from about 1×10 6 to about 1×10 10 colony forming units (CFU) of said at least one porcine lactic acid bacterial strain per gram of the composition.

33 . The pharmaceutical composition of claim 1 , in unit dose form.

34 . The pharmaceutical composition of claim 1 , wherein the intestinal disorder is selected from the group consisting of salmonellosis, irritable bowel syndrome (MS), inflammatory bowel disorder (IBD), functional dyspepsia, functional constipation, func

Assignees

• 4D Pharma Res Ltd

Inventors

• Kelly Denise

Classifications

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• A23K10/18

  of live microorganisms · CPC title

• A23V2002/00

  Food compositions, function of food ingredients or processes for food or foodstuffs · CPC title

• A23L33/135

  Bacteria or derivatives thereof, e.g. probiotics · CPC title