Inhibition and treatment of gastrointestinal biofilms

Abstract

Orally administered physiologically acceptable anti-biofilm compositions comprising enzymes and if desired additional components such as antimicrobials, antibiotics, antifungals, herbals, chelating agents, lactoferrin and related compounds, minerals, surfactants, binders, and fillers useful for the inhibition and treatment of gastrointestinal biofilms in humans. Physiologically acceptable anti-biofilm compositions containing these enzymes are useful in the inhibition, reduction and/or treatment of gastrointestinal biofilm infections, and associated systemic symptoms caused by biofilms associated microorganisms within the gastrointestinal tract. Methods of identification, preparation and use of such physiologically acceptable anti-biofilm compositions are also provided.

1 . A physiologically acceptable anti-biofilm composition suitable for oral administration to a mammal while retaining effectiveness in the gut, the composition comprising at least one of an anti-biofilm acid-stable cellulase or an anti-biofilm anti-polymeric β-1,6-N-acetyl-D-glucosamine (poly-β-1,6-GlcNAc) agent in an amount capable of significant biofilm degradation in at least one pharmaceutically acceptable carrier. 
   
   
       2 . The composition of  claim 1  wherein the composition further comprises both the anti-biofilm acid-stable cellulase and the anti-biofilm anti-poly-β-1,6-GlcNAc agent. 
   
   
       3 . The composition of  claim 1  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is a hexosaminidase. 
   
   
       4 . The composition of  claim 1  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is Dispersin B. 
   
   
       5 . The composition of  claim 1  wherein the composition further comprises an effective amount of at least one of an acid-stable hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, or  Serratia  peptidase. 
   
   
       6 . The composition of  claim 5  wherein the composition further comprises at least three of the hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, and  Serratia  peptidase. 
   
   
       7 . The composition of  claim 6  wherein the composition further comprises all of hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, and  Serratia  peptidase. 
   
   
       8 . The composition of  claim 5  wherein the amount of cellulase per dose is about 100-300 CU, the amount of hemicellulase/pectinase complex is about 60-100 HSU, the amount of β-gluconase is about 6-10 BGU, the amount of acid protease is about 15-25 SAP, and the amount of alkaline protease is about 15-25 HUT. 
   
   
       9 . The composition of  claim 5  wherein the amount of cellulase per dose is about 200 CU, the amount of hemicellulase/pectinase complex is about 80 HSU, the amount of β-gluconase is about 8 BGU, the amount of acid protease is about 20 SAP, and the amount of alkaline protease is about 20 HUT. 
   
   
       10 . The composition of  claim 5  wherein the amount of cellulase per dose ranges from 1 to 10,000 CU, the amount of hemicellulase/pectinase complex ranges from 1 to 8,000 HSU, the amount of β-gluconase ranges from 1 to 1000 BGU, the amount of acid protease ranges from 1 to 10,000 SAP, and the amount of alkaline protease ranges from 1 to 40,000 HUT. 
   
   
       11 . The composition of  claim 1  wherein the physiologically acceptable anti-biofilm composition further comprises an effective amount of at least one an acid-stable agent, the at least one agent selected from the following: a disaccharide; amylase; α-amylase; β-amylase; glucoamylase; endoglucanase; xylanase; lipase; lysozyme; an enzyme with dipeptidyl peptidase IV (DPP-IV) activity; chitosanase; bromelain; papain; ficin; kiwi protease; any plant-derived protease or proteinase, or phytase. 
   
   
       12 . The composition of  claim 11  wherein the lipase is a microbial lipase. 
   
   
       13 . The composition of  claim 11  wherein the lipase comprises a lipase from at least one of  Candida, Pseudomonas, Bacillus, Humicola  or  Rhizomucor.    
   
   
       14 . The composition of  claim 11  wherein the amylase is at least one of a  Bacillus  amylase or  Aspergillus  amylase. 
   
   
       15 . The composition of  claim 11  wherein the composition further comprises at least one pectinase that is at least one of a polygalacturonase (EC3.2.1.15), pectinesterase (EC3.2.1.11), pectin lyase (EC4.2.2.10) or hemicellulase. 
   
   
       16 . The composition of  claim 11  wherein the pectinase is at least one an  Aspergillus niger  pectinase or  Aspergillus  aculeatus pectinase. 
   
   
       17 . The composition of  claim 1  wherein the physiologically acceptable anti-biofilm composition further comprises at least one acid-stable enzyme in an amount capable of biofilm degradation, the at least one enzyme selected from the following: 1,2-1,3-α-D-mannan mannohydrolase, 1,3-β-D-xylanxylanohydrolase, 1,3-β-D-glucan glucanohydrolase, 1,3(1,3; 1,4)-α-D-glucan 3-glucanohydrolase, 1,3(1,3; 1,4)-β-D-glucan 3(4)-glucanohydrolase, 1,3-1,4-α-D-glucan 4-glucanohydrolase, 1,4-α-D-glucan glucanehydrolase, 1,4-α-D-glucan glucohydrolase, 1,4-(1,3:1,4)-β-D-glucan 4-glucanohydrolase, 1,4-β-D-glucan glucohydrolase, 1,4-β-D-xylan xylanohydrolase, 1,4-β-D-mannan mannanohydrolase, 1,5-α-L-arabinanohydrolase, 1,4-α-D-glucan maltohydrolase, 1,6-α-D-glucan 6-glucanohydrolase, 2,6-β-fructan fructanohydrolase, α-dextrin 6-glucanohydrolase, α-D-galactoside galactohydrolase, α-D-glucoside glucohydrolase, α-D-mannoside mannohydrolase, acylneuraminyl hydrolase, Aerobacter-capsular-polysaccharide galactohydrolase, β-D-fructofuranoside fructohydrolase, 1-D-fucoside fucohydrolase, α-D-fructan fructohydrolase, β-D-galactoside galactohydrolase, 13-D-glucoside glucohydrolase, β-D-glucuronoside, glucuronosohydrolase, β-D-mannoside mannohydrolase, β-N-acetyl-D-hexosaminide N-acetylhexosamino hydrolase, cellulose-sulfate sulfohydrolase, collagenase, dextrin 6-α-D-glucanohydrolase, glycoprotein-phosphatidylinositol phosphatidohydrolase, hyaluronate 4-glycanohydrolase, hyaluronoglucuronidase, pectin pectylhydrolase, peptidoglycan N-acetylmuramoylhydrolase, phosphatidylcholine 2-acylhydrolase, phosphatidylcholine 1-acylhydrolase, poly(1,4-α-D-galacturonide), poly(1,4-(N-acetyl-β-D-glucosaminide))-glycanohydrolase, proteases, sucrose α-glucosidase, triacylglycerol acylhydrolase, triacylglycerol protein-acylhydrolase. 
   
   
       18 . The composition of  claim 1  wherein the composition further comprises an acid-stable subtilisin in an amount capable of biofilm degradation. 
   
   
       19 . The composition of  claim 1 , further comprising acid-stable DNAse I in an amount capable of biofilm degradation. 
   
   
       20 . The composition of  claim 1  wherein the composition further comprises at least one of oil of oregano, berberine, undecylenic acid, a prescription antibiotic, a prescription antimicrobial, a probiotic microorganism or a prebiotic. 
   
   
       21 . A method of screening for a physiologically acceptable anti-biofilm composition suitable for oral administration to a mammal while retaining effectiveness in the gut, the method comprising,
 providing a significant plurality of samples of a live target biofilm on at least one substrate;   applying to each of the plurality of samples one of range of doses of a candidate anti-biofilm agent selected from the group comprising acid-stable cellulase and an anti-biofilm anti-polymeric β-1,6-N-acetyl-D-glucosamine (poly-β-1,6-GlcNAc) agent, under conditions wherein the samples of the target biofilm can grow absent a significant anti-biofilm effect due to the candidate anti-biofilm agent; and,   determining whether each of the range of doses of candidate anti-biofilm agent inhibited growth of its respective sample.   
   
   
       22 . The method of  claim 21  wherein the method further comprises screening both the anti-biofilm acid-stable cellulase and the anti-biofilm anti-poly-β-1,6-GlcNAc agent. 
   
   
       23 . The method of  claim 21  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is a hexosaminidase. 
   
   
       24 . The method of  claim 21  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is Dispersin B. 
   
   
       25 . The method of  claim 21  wherein the method further comprises screening at least one of an acid-stable hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, or  Serratia  peptidase. 
   
   
       26 . The method of  claim 25  wherein the amount of cellulase is equivalent to a dose of about 100-300 CU, the amount of hemicellulase/pectinase complex is about 60-100 HSU, the amount of β-gluconase is about 6-10 BGU, the amount of acid protease is about 15-25 SAP, and the amount of alkaline protease is about 15-25 HUT. 
   
   
       27 . The method of  claim 21  wherein the method further comprises screening at least one an acid-stable agent selected from the following: a disaccharide; amylase; α-amylase; β-amylase; glucoamylase; endoglucanase; xylanase; lipase; lysozyme; an enzyme with dipeptidyl peptidase IV (DPP-IV) activity; chitosanase; bromelain; papain; ficin; kiwi protease; any plant-derived protease or proteinase, or phytase. 
   
   
       28 . The method of  claim 27  wherein the lipase is a microbial lipase. 
   
   
       29 . The method of  claim 27  wherein the lipase comprises a lipase from at least one of  Candida, Pseudomonas, Bacillus, Humicola  or  Rhizomucor.    
   
   
       30 . The method of  claim 27  wherein the amylase is at least one of a  Bacillus  amylase or  Aspergillus  amylase. 
   
   
       31 . The method of  claim 27  wherein the method comprises screening at least one pectinase that is at least one of a polygalacturonase (EC3.2.1.15), pectinesterase (EC3.2.1.11), pectin lyase (EC4.2.2.10) or hemicellulase. 
   
   
       32 . The method of  claim 27  wherein the pectinase is at least one an  Aspergillus niger  pectinase or  Aspergillus aculeatus  pectinase. 
   
   
       33 . The method of  claim 21  wherein the method further comprises screening at least one of the following: 1,2-1,3-α-D-mannan mannohydrolase, 1,3-β-D-xylanxylanohydrolase, 1,3-β-D-glucan glucanohydrolase, 1,3(1,3; 1,4)-α-D-glucan 3-glucanohydrolase, 1,3(1,3; 1,4)-β-D-glucan 3(4)-glucanohydrolase, 1,3-1,4-α-D-glucan 4-glucanohydrolase, 1,4-α-D-glucan glucanehydrolase, 1,4-α-D-glucan glucohydrolase, 1,4-(1,3:1,4)-β-D-glucan 4-glucanohydrolase, 1,4-β-D-glucan glucohydrolase, 1,4-β-D-xylan xylanohydrolase, 1,4-β-D-mannan mannanohydrolase, 1,5-α-L-arabinanohydrolase, 1,4-α-D-glucan maltohydrolase, 1,6-α-D-glucan 6-glucanohydrolase, 2,6-β-fructan fructanohydrolase, α-dextrin 6-glucanohydrolase, α-D-galactoside galactohydrolase, α-D-glucoside glucohydrolase, α-D-mannoside mannohydrolase, acylneuraminyl hydrolase, Aerobacter-capsular-polysaccharide galactohydrolase, β-D-fructofuranoside fructohydrolase, β-D-fucoside fucohydrolase, α-D-fructan fructohydrolase, β-D-galactoside galactohydrolase, β-D-glucoside glucohydrolase, β-D-glucuronoside, glucuronosohydrolase, β-D-mannoside mannohydrolase, β-N-acetyl-D-hexosaminide N-acetylhexosamino hydrolase, cellulose-sulfate sulfohydrolase, collagenase, dextrin 6-α-D-glucanohydrolase, glycoprotein-phosphatidylinositol phosphatidohydrolase, hyaluronate 4-glycanohydrolase, hyaluronoglucuronidase, pectin pectylhydrolase, peptidoglycan N-acetylmuramoylhydrolase, phosphatidylcholine 2-acylhydrolase, phosphatidylcholine 1-acylhydrolase, poly(1,4-α-D-galacturonide), poly(1,4-(N-acetyl-β-D-glucosaminide))-glycanohydrolase, proteases, sucrose α-glucosidase, triacylglycerol acylhydrolase, triacylglycerol protein-acylhydrolase. 
   
   
       34 . The method of  claim 21  wherein the method further comprises screening an acid-stable subtilisin. 
   
   
       35 . The method of  claim 21  wherein the method further comprises screening an acid-stable DNAse I. 
   
   
       36 . The method of  claim 21  wherein the method further comprises screening at least one of oil of oregano, berberine, undecylenic acid, a prescription antibiotic, a prescription antimicrobial, a probiotic microorganism or a prebiotic. 
   
   
       37 . A method of inhibiting a gastrointestinal biofilm infection in a mammal, the method comprising:
 identifying the presence of the gastrointestinal biofilm infection,   orally administering to the mammal a therapeutically effective amount of at least one anti-biofilm agent comprising an acid-stable cellulase or an anti-polymeric β-1,6-N-acetyl-D-glucosamine (poly-β-1,6-GlcNAc) agent in at least one pharmaceutically acceptable carrier, in an amount and for a time sufficient to cause significant biofilm degradation within the gastrointestinal system of the mammal.   
   
   
       38 . The method of  claim 37  wherein the method further comprises administering both the anti-biofilm acid-stable cellulase and the anti-biofilm anti-poly-β-1,6-GlcNAc agent. 
   
   
       39 . The method of  claim 37  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is a hexosaminidase. 
   
   
       40 . The method of  claim 37  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is Dispersin B. 
   
   
       41 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of at least one of an acid-stable hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, or  Serratia  peptidase. 
   
   
       42 . The method of  claim 41  wherein the amount of cellulase per dose is about 100-300 CU, the amount of hemicellulase/pectinase complex is about 60-100 HSU, the amount of β-gluconase is about 6-10 BGU, the amount of acid protease is about 15-25 SAP, and the amount of alkaline protease is about 15-25 HUT. 
   
   
       43 . The method of  claim 41  wherein the amount of cellulase per dose ranges from 1 to 10,000 CU, the amount of hemicellulase/pectinase complex ranges from 1 to 8,000 HSU, the amount of β-gluconase ranges from 1 to 1000 BGU, the amount of acid protease ranges from 1 to 10,000 SAP, and the amount of alkaline protease ranges from 1 to 40,000 HUT. 
   
   
       44 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of at least one an acid-stable agent selected from the following: a disaccharide; amylase; α-amylase; β-amylase; glucoamylase; endoglucanase; xylanase; lipase; lysozyme; an enzyme with dipeptidyl peptidase IV (DPP-IV) activity; chitosanase; bromelain; papain; ficin; kiwi protease; any plant-derived protease or proteinase, or phytase. 
   
   
       45 . The method of  claim 44  wherein the lipase is a microbial lipase. 
   
   
       46 . The method of  claim 44  wherein the lipase comprises a lipase from at least one of  Candida, Pseudomonas, Bacillus, Humicola  or  Rhizomucor.    
   
   
       47 . The method of  claim 44  wherein the amylase is at least one of a  Bacillus  amylase or  Aspergillus  amylase. 
   
   
       48 . The method of  claim 44  wherein the method further comprises administering a therapeutically effective amount of at least one pectinase that is at least one of a polygalacturonase (EC3.2.1.15), pectinesterase (EC3.2.1.11), pectin lyase (EC4.2.2.10) or hemicellulase. 
   
   
       49 . The method of  claim 44  wherein the pectinase is at least one an  Aspergillus niger  pectinase or  Aspergillus  aculeatus pectinase. 
   
   
       50 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of at least one acid-stable enzyme selected from the following: 1,2-1,3-α-D-mannan mannohydrolase, 1,3-β-D-xylanxylanohydrolase, 1,3-β-D-glucan glucanohydrolase, 1,3(1,3; 1,4)-α-D-glucan 3-glucanohydrolase, 1,3(1,3; 1,4)-β-D-glucan 3(4)-glucanohydrolase, 1,3-1,4-α-D-glucan 4-glucanohydrolase, 1,4-α-D-glucan glucanehydrolase, 1,4-α-D-glucan glucohydrolase, 1,4-(1,3:1,4)-β-D-glucan 4-glucanohydrolase, 1,4-β-D-glucan glucohydrolase, 1,4-β-D-xylan xylanohydrolase, 1,4-β-D-mannan mannanohydrolase, 1,5-α-L-arabinanohydrolase, 1,4-α-D-glucan maltohydrolase, 1,6-α-D-glucan 6-glucanohydrolase, 2,6-β-fructan fructanohydrolase, α-dextrin 6-glucanohydrolase, α-D-galactoside galactohydrolase, α-D-glucoside glucohydrolase, α-D-mannoside mannohydrolase, acylneuraminyl hydrolase, Aerobacter-capsular-polysaccharide galactohydrolase, β-D-fructofuranoside fructohydrolase, β-D-fucoside fucohydrolase, α-D-fructan fructohydrolase, β-D-galactoside galactohydrolase, β-D-glucoside glucohydrolase, β-D-glucuronoside, glucuronosohydrolase, β-D-mannoside mannohydrolase, β-N-acetyl-D-hexosaminide N-acetylhexosamino hydrolase, cellulose-sulfate sulfohydrolase, collagenase, dextrin 6-α-D-glucanohydrolase, glycoprotein-phosphatidylinositol phosphatidohydrolase, hyaluronate 4-glycanohydrolase, hyaluronoglucuronidase, pectin pectylhydrolase, peptidoglycan N-acetylmuramoylhydrolase, phosphatidylcholine 2-acylhydrolase, phosphatidylcholine 1-acylhydrolase, poly(1,4-α-D-galacturonide), poly(1,4-(N-acetyl-β-D-glucosaminide))-glycanohydrolase, proteases, sucrose α-glucosidase, triacylglycerol acylhydrolase, triacylglycerol protein-acylhydrolase. 
   
   
       51 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of an acid-stable subtilisin. 
   
   
       52 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of an acid-stable DNAse I. 
   
   
       53 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of at least one of oil of oregano, berberine, undecylenic acid, a prescription antibiotic, a prescription antimicrobial, a probiotic microorganism or a prebiotic. 
   
   
       54 . The composition of  claim 1  for use as an active therapeutic substance. 
   
   
       55 . The composition of  claim 1  for use in the manufacture of a medicament for inhibiting or treating a gastrointestinal biofilm in a mammal. 
   
   
       56 . A method of manufacturing a medicament able to reduce symptoms associated with a gastrointestinal biofilm in a human patient, comprising combining a pharmaceutically effective amount of at least one of an anti-biofilm acid-stable cellulase or an anti-biofilm anti-polymeric β-1,6-N-acetyl-D-glucosamine (poly-β-1,6-GlcNAc) agent in an amount capable of significant biofilm degradation with at least one of a pharmaceutically acceptable carrier, adjuvant, excipient, buffer and diluent. 
   
   
       57 . The method of  claim 56  wherein the medicament further comprises both the anti-biofilm acid-stable cellulase and the anti-biofilm anti-poly-β-1,6-GlcNAc agent. 
   
   
       58 . The method of  claim 56  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is a hexosaminidase. 
   
   
       59 . The method of  claim 56  wherein the anti-biofilm anti-poly-β-1,6-GlcNAc agent is Dispersin B. 
   
   
       60 . The method of  claim 56  wherein the medicament further comprises an effective amount of at least one of an acid-stable hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, or  Serratia  peptidase. 
   
   
       61 . The method of  claim 60  wherein the medicament further comprises at least three of the hemicellulase/pectinase complex, 6-gluconase, acid protease, alkaline protease, and  Serratia  peptidase. 
   
   
       62 . The method of  claim 60  wherein the medicament further comprises all of hemicellulase/pectinase complex, β-gluconase, acid protease, alkaline protease, and  Serratia  peptidase. 
   
   
       63 . The method of  claim 60  wherein the amount of cellulase per dose is about 100-300 CU, the amount of hemicellulase/pectinase complex is about 60-100 HSU, the amount of β-gluconase is about 6-10 BGU, the amount of acid protease is about 15-25 SAP, and the amount of alkaline protease is about 15-25 HUT. 
   
   
       64 . The method of  claim 60  wherein the amount of cellulase per dose is about 200 CU, the amount of hemicellulase/pectinase complex is about 80 HSU, the amount of β-gluconase is about 8 BGU, the amount of acid protease is about 20 SAP, and the amount of alkaline protease is about 20 HUT. 
   
   
       65 . The method of  claim 60  wherein the amount of cellulase per dose ranges from 1 to 10,000 CU, the amount of hemicellulase/pectinase complex ranges from 1 to 8,000 HSU, the amount of β-gluconase ranges from 1 to 1000 BGU, the amount of acid protease ranges from 1 to 10,000 SAP, and the amount of alkaline protease ranges from 1 to 40,000 HUT. 
   
   
       66 . The method of  claim 56  wherein the physiologically acceptable anti-biofilm medicament further comprises an effective amount of at least one an acid-stable agent, the at least one agent selected from the following: a disaccharide; amylase; α-amylase; β-amylase; glucoamylase; endoglucanase; xylanase; lipase; lysozyme; an enzyme with dipeptidyl peptidase IV (DPP-IV) activity; chitosanase; bromelain; papain; ficin; kiwi protease; any plant-derived protease or proteinase, or phytase. 
   
   
       67 . The method of  claim 66  wherein the lipase is a microbial lipase. 
   
   
       68 . The method of  claim 66  wherein the lipase comprises a lipase from at least one of  Candida, Pseudomonas, Bacillus, Humicola  or  Rhizomucor.    
   
   
       69 . The method of  claim 66  wherein the amylase is at least one of a  Bacillus  amylase or  Aspergillus  amylase. 
   
   
       70 . The method of  claim 66  wherein the medicament further comprises at least one pectinase that is at least one of a polygalacturonase (EC3.2.1.15), pectinesterase (EC3.2.1.11), pectin lyase (EC4.2.2.10) or hemicellulase. 
   
   
       71 . The method of  claim 66  wherein the pectinase is at least one an  Aspergillus niger  pectinase or  Aspergillus aculeatus  pectinase. 
   
   
       72 . The method of  claim 56  wherein the physiologically acceptable anti-biofilm medicament further comprises at least one acid-stable enzyme in an amount capable of biofilm degradation, the at least one enzyme selected from the following: 1,2-1,3-α-D-mannan mannohydrolase, 1,3-β-D-xylanxylanohydrolase, 1,3-β-D-glucan glucanohydrolase, 1,3(1,3; 1,4)-α-D-glucan 3-glucanohydrolase, 1,3(1,3; 1,4)-β-D-glucan 3(4)-glucanohydrolase, 1,3-1,4-α-D-glucan 4-glucanohydrolase, 1,4-α-D-glucan glucanehydrolase, 1,4-α-D-glucan glucohydrolase, 1,4-(1,3:1,4)-β-D-glucan 4-glucanohydrolase, 1,4-β-D-glucan glucohydrolase, 1,4-β-D-xylan xylanohydrolase, 1,4-β-D-mannan mannanohydrolase, 1,5-α-L-arabinanohydrolase, 1,4-α-D-glucan maltohydrolase, 1,6-α-D-glucan 6-glucanohydrolase, 2,6-β-fructan fructanohydrolase, α-dextrin 6-glucanohydrolase, α-D-galactoside galactohydrolase, α-D-glucoside glucohydrolase, α-D-mannoside mannohydrolase, acylneuraminyl hydrolase, Aerobacter-capsular-polysaccharide galactohydrolase, β-D-fructofuranoside fructohydrolase, β-D-fucoside fucohydrolase, α-D-fructan fructohydrolase, β-D-galactoside galactohydrolase, β-D-glucoside glucohydrolase, β-D-glucuronoside, glucuronosohydrolase, β-D-mannoside mannohydrolase, β-N-acetyl-D-hexosaminide N-acetylhexosamino hydrolase, cellulose-sulfate sulfohydrolase, collagenase, dextrin 6-α-D-glucanohydrolase, glycoprotein-phosphatidylinositol phosphatidohydrolase, hyaluronate 4-glycanohydrolase, hyaluronoglucuronidase, pectin pectylhydrolase, peptidoglycan N-acetylmuramoylhydrolase, phosphatidylcholine 2-acylhydrolase, phosphatidylcholine 1-acylhydrolase, poly(1,4-α-D-galacturonide), poly(1,4-(N-acetyl-β-D-glucosaminide))-glycanohydrolase, proteases, sucrose α-glucosidase, triacylglycerol acylhydrolase, triacylglycerol protein-acylhydrolase. 
   
   
       73 . The method of  claim 56  wherein the medicament further comprises an acid-stable subtilisin in an amount capable of biofilm degradation. 
   
   
       74 . The method of  claim 56  further comprising acid-stable DNAse I in an amount capable of biofilm degradation. 
   
   
       75 . The method of  claim 56  wherein the medicament further comprises at least one of oil of oregano, berberine, undecylenic acid, a prescription antibiotic, a prescription antimicrobial, a probiotic microorganism or a prebiotic. 
   
   
       76 . The composition of  claim 1  wherein the composition further comprises an effective amount of a  Serratia  peptidase. 
   
   
       77 . The method of  claim 21  wherein the method further comprises screening a  Serratia  peptidase. 
   
   
       78 . The method of  claim 37  wherein the method further comprises administering a therapeutically effective amount of a  Serratia  peptidase. 
   
   
       79 . The method of  claim 56  wherein the medicament further comprises an effective amount of a  Serratia  peptidase. 
   
   
       80 . A physiologically acceptable anti-biofilm composition suitable for oral administration to a mammal while retaining effectiveness in the gut, the composition comprising at least one of an anti-biofilm  Serratia  peptidase agent in an amount capable of significant biofilm degradation in at least one pharmaceutically acceptable carrier. 
   
   
       81 . A method of screening for a physiologically acceptable anti-biofilm composition suitable for oral administration to a mammal while retaining effectiveness in the gut, the method comprising,
 providing a significant plurality of samples of a live target biofilm on at least one substrate;   applying to each of the plurality of samples one of range of doses of a candidate anti-biofilm agent comprising a  Serratia  peptidase agent, under conditions wherein the samples of the target biofilm can grow absent a significant anti-biofilm effect due to the candidate anti-biofilm agent; and,   determining whether each of the range of doses of candidate anti-biofilm agent inhibited growth of its respective sample.   
   
   
       82 . A method of inhibiting a gastrointestinal biofilm infection in a mammal, the method comprising:
 identifying the presence of the gastrointestinal biofilm infection,   orally administering to the mammal a therapeutically effective amount of at least one anti-biofilm agent comprising a  Serratia  peptidase agent in at least one pharmaceutically acceptable carrier, in an amount and for a time sufficient to cause significant biofilm degradation within the gastrointestinal system of the mammal.   
   
   
       83 . A method of manufacturing a medicament able to reduce symptoms associated with a gastrointestinal biofilm in a human patient, comprising combining a pharmaceutically effective amount of a  Serratia  peptidase agent in an amount capable of significant biofilm degradation with at least one of a pharmaceutically acceptable carrier, adjuvant, excipient, buffer and diluent.