Method of identifying a biologically-active composition from a biofilm
Inventors
Berkes, Eva A. • Monsul, Nicholas T.
Assignees
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Publication Number
US-9504739-B2
Publication Date
2016-11-29
Expiration Date
Abstract
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.
Core Innovation
The invention provides a method for identifying and selecting a biologically-active composition by growing a microbial biofilm on a substrate contained within a reactor vessel and contacted with a biological growth medium in the reactor vessel. The biofilm is harvested by removing the biofilm from the substrate and separating the biofilm from the growth medium to obtain a fraction comprising biofilm cellular mass. This fraction is tested for biological activity and then selected as the biologically-active composition when the fraction is found to have biological activity.
The biological activity determined in the method is anti-microbial activity or anti-inflammatory activity. The approach is aimed at preventing, dispersing, and/or treating pathogenic biofilms and associated chronic inflammation by using natural, non-toxic compositions that include one or more probiotic organisms or fractions and metabolites. The compositions are also described as supporting innate immunity and microbiotic homeostasis while targeting pathogenic biofilms.
In the described implementations, the method and compositions are associated with mechanisms such as preventing or disrupting deposition, inhibiting or disrupting EPS matrix secretion, and preventing, inhibiting, or disrupting quorum sensing. The document further describes biofilm efficacy testing using MBEC/BIC approaches, including biofilm growth on glass spheres with shear in a rotating biofilm reactor and evaluation against planktonic MIC comparisons, along with fractionation and removal from biofilm material.
Claims Coverage
The independent claim covers a method that identifies and selects a biologically-active composition from fractions of harvested microbial biofilm cellular mass, where the biological activity is anti-microbial and/or anti-inflammatory, determined by testing the harvested fraction.
Reactor-based biofilm growth and harvesting fraction
Growing a microbial biofilm on a substrate contained within a reactor vessel and contacted with a biological growth medium; harvesting the biofilm by removing it from the substrate and separating the biofilm from the growth medium to obtain a fraction comprising biofilm cellular mass.
Biofilm fraction biological activity testing and selection
Testing the fraction of biofilm cellular mass for biological activity; determining whether the fraction has biological activity; selecting, as the biologically-active composition, the fraction that is found to have biological activity, wherein the biological activity is anti-microbial activity or anti-inflammatory activity.
Anti-microbial testing via attachment inhibition, matrix disruption, and quorum-sensing interference
Testing the fraction for one or more anti-microbial properties against biofilm by evaluating attachment inhibition, matrix disruption, and quorum-sensing prevention, inhibition, or disruption.
Biofilm cultivation under applied shear
Growing a biofilm under applied shear stress in the method of claim 1.
Sonication-based biofilm removal
Removing biofilm from a substrate using sonication.
Centrifugation or filtration fraction separation
Separating the supernatant from the cell mass using centrifugation or filtration.
Probiotic microbe selection from specified genera
Selecting the probiotic microbe from Bacteroides, Bifidobacterium, or Lactobacillus.
Overall, the claim set centers on selecting an active composition by reactor-based growth of microbial biofilms, harvesting and fractionating biofilm cellular mass, and testing for anti-microbial activity and/or anti-inflammatory activity, with dependents narrowing biofilm production and removal conditions and specifying anti-microbial testing mechanisms such as attachment inhibition, EPS matrix disruption, and quorum-sensing prevention/inhibition/disruption.
Stated Advantages
Prevents, disperses, and/or treats pathogenic biofilms and associated chronic inflammation.
Supports innate immunity and microbiotic homeostasis.
Provides a biologically-active composition via selecting a fraction found to have anti-microbial and/or anti-inflammatory biological activity.
Documented Applications
Ocular dry eye/dry irritated eyes, including meibomian gland dysfunction, associated with biofilm-related inflammation.
Chronic rhinosinusitis associated with biofilm-related inflammatory conditions.
Periodontitis and chronic periodontitis associated with pathogenic biofilms and inflammation.
Respiratory biofilm-related inflammatory conditions.
Dermatologic biofilm-related inflammatory conditions.
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