Method for Identifying Pre-Biotics and Compositions Containing the Same
A method for indentifying test agents that exhibit prebiotic activity on companion animal skin commensal microorganisms and dermatological compositions that include such agents. The method includes providing a test culture of a test agent, a companion animal skin commensal microorganism and a minimal carbon medium. The method provides a time efficient and cost effective way to predict in vivo prebiotic activity of a test agent on skin commensal microorganisms.
1 . A method for identifying a test agent as a prebiotic, the method comprising:
providing a first culture comprising a test agent, a quantity of a companion animal skin commensal microorganism, and a minimal carbon media;
measuring a metabolite level or a replication level of the companion animal skin commensal microorganism of the first culture;
comparing the measured metabolite level or replication level to a control value; and
identifying the test agent as a prebiotic when the measured replication level or the measured metabolic level is greater than the control value.
2 . The method of claim 1 , wherein the providing the first culture comprises combining the test agent, the companion animal skin commensal microorganism, and the minimal carbon media in a vessel.
3 . The method of claim 2 , wherein an amount of time passes prior to measuring the metabolite level or replication level.
4 . The method of claim 3 , wherein the test agent induces a change in the companion animal skin commensal microorganism during the amount of time that passes.
5 . The method of claim 4 , wherein the at least one of the metabolite level and the replication level changes.
6 . The method of claim 5 , wherein the change is an increase in metabolite level and the metabolite level is measured with an ATP assay.
7 . A method for identifying a test agent as a prebiotic, the method comprising:
combining a test agent, a quantity of a companion animal skin commensal microorganism, and a minimal carbon media in a vessel to provide a first culture;
measuring a metabolite level or a replication level of the companion animal skin commensal microorganism of the first culture;
comparing the measured metabolite level or replication level to a control value; and
identifying the test agent as a prebiotic when the measured replication level or the measured metabolic level is greater than the control value.
8 . The method of claim 7 , wherein the companion animal skin commensal microorganism is a species from the genus Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Propionibacteria, Corynebacteria, Actinobacteria, Clostridiales, Lactobacillales, Staphylococcus, Bacillus, Micrococcus, Streptococcus, Bacteroidales, Flavobacteriales, Enterococcus, Pseudomonas, Malassezia, Maydida, Debaroyomyces , or.
9 . The method of claim 7 , wherein the first culture includes more than one kind of companion animal skin commensal microorganisms.
10 . The method of claim 7 , wherein the companion animal skin commensal microorganism comprises a reporter gene.
11 . The method of claim 7 , wherein the minimal carbon media has a carbon content of less than about 0.001% by weight, based on the weight of the media.
12 . The method of claim 7 , wherein the minimal carbon media comprises NaCl, (NH4)2HPO4, K2HPO4, and MgSO4.7H2O.
13 . The method of claim 7 , wherein the quantity of the companion animal skin commensal microorganism in the first culture is between about 0.5×10 7 CFU/ml and 5×10 7 CFU/ml
14 . The method of claim 7 , wherein the first culture has a pH of between about 6.6 and about 7.4.
15 . A dermatological composition, formulated for topical application to a body surface, comprising a dermatologically acceptable carrier and at least one test agent identified as a prebiotic by the method of claim 7 .
16 . The method of claim 7 , wherein the control value is obtained by measuring the metabolite level or replication level of a second culture.
17 . The method of claim 7 , wherein the second culture comprises the companion animal skin commensal microorganism and a minimal carbon media and wherein the second culture is substantially free of the test agent.
18 . The method of claim 7 , wherein the control is substantially free of iron, biotin, nicotinic acid, D-pantothenic acid, pyridoxal, pyridoxamine dihydrochloride, thiamin hydrochloride, valine, arginine, galactose, mannose, fructose, sucrose, lactose, and maltose.
19 . The method of claim 7 , wherein the metabolite level is measured by measuring an amount of ATP present in the first culture.
20 . The method of claim 19 , wherein the control value is obtained by measuring an amount of ATP present in a second culture comprising the companion animal skin commensal microorganism and wherein the second culture is substantially free of the test agent.