Method of identifying a biologically-active composition from a biofilm
The subject invention provides materials and methods that effectively support innate immunity and/or disperse pathogenic biofilms using readily available, nontoxic, natural substances, while supporting restoration of normal microbiotic homeostasis. In one embodiment, the subject invention provides anti-biofilm compositions comprising one or more probiotic organisms, anti-microbial honey, and other ingredients such as prebiotic compounds, other hive products, green tea derivatives, other plant derivatives, and vitamin D3.
1. A method for identifying and selecting a biologically-active composition, wherein said method comprises: (1) growing a microbial biofilm on a substrate wherein said substrate upon which said biofilm is grown is contained within a reactor vessel and is contacted with a biological growth medium within said reactor vessel; (2) harvesting said biofilm by removing the biofilm from the substrate and separating the biofilm from the growth medium thereby obtaining a fraction comprising biofilm cellular mass; (3) testing said fraction of biofilm cellular mass for biological activity; (4) based on results of the testing in step (3), determining whether said fraction has biological activity; and (5) selecting, as the biologically-active composition, the fraction that is found to have biological activity; wherein said biological activity is anti-microbial activity or anti-inflammatory activity.
2. The method, according to claim 1 , wherein the biofilm is of a probiotic microbe.
3. The method, according to claim 1 , wherein the biofilm is grown from a biological sample obtained from a human.
4. The method, according to claim 1 , wherein the biological activity is anti-microbial activity.
5. The method, according to claim 1 , wherein the biological activity is anti-inflammatory activity.
6. The method, according to claim 1 , wherein the fraction is tested for one or more anti-microbial properties against biofilm selected from the group consisting of attachment inhibition, matrix disruption, and preventing, inhibiting, or disrupting quorum-sensing mechanisms.
7. The method, according to claim 2 , wherein the probiotic microbe is selected from the group consisting of Bacteroides, Bifidobacterium , and Lactobacillus.
8. The method, according to claim 7 , wherein the microbe is a Lactobacillus.
9. The method, according to claim 1 , wherein said method comprises removing the biofilm from the substrate by sonication.
10. The method, according to claim 9 , wherein the substrate comprises glass.
11. The method, according to claim 1 , wherein the biofilm is grown under shear stress.
12. The method, according to claim 1 , wherein the step of separating the supernatant from the cell mass comprises centrifugation or filtration.
13. The method, according to claim 12 , wherein centrifugation is used to separate the supernatant from the cell mass.