Biofilm Sampling Composition for Viable Microorganism Collection

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Solution Overview

Problem

Conventional methods for removing and collecting microorganisms, particularly those in biofilm form, are ineffective due to the protective extracellular matrix, leading to non-specific treatments, incomplete collection, and interference with downstream microbiological tests, especially for sensitive bacteria like Listeria.

Innovation Solution

A composition comprising anionic surfactants, enzymes (protease, polysaccharidase, laccase, lipase, cellulase, mannanase), polyol stabilizers, and sequestering agents, formulated to degrade biofilms without interfering with downstream tests, ensuring representative sampling and viability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional biocides are used to eliminate microorganisms, then microorganism removal is attempted, but the extracellular matrix of biofilms provides resistance and protects microorganisms, making elimination very difficult

Engineering Contradiction:
Improvemicroorganism elimination effectivenessVSAvoidbiofilm resistance
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention divides the biofilm removal function into multiple enzymatic components, each targeting specific parts of the extracellular matrix (proteases for proteins, polysaccharidases for polysaccharides, lipases for lipids, cellulases for cellulose, mannanases for mannans). This segmentation allows comprehensive degradation of the complex matrix structure that single biocides cannot penetrate or destroy.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention uses enzymes as intermediary substances to mediate the breakdown of the extracellular matrix. These enzymes act as biological mediators that specifically degrade the protective matrix components, enabling microorganism removal without direct contact between biocides and the protected microorganisms.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Adaptability or versatility

If broad-spectrum biocides are used to treat diverse microorganisms, then coverage is increased, but non-specific treatments cause interference with downstream microbiological tests

Engineering Contradiction:
Improvemicroorganism coverageVSAvoiddownstream test accuracy
Core Design Contradiction:
Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The invention extracts and removes only the extracellular matrix components using specific enzymes, leaving the microorganisms themselves intact and free from contaminating chemical residues. This selective extraction allows downstream microbiological tests to accurately detect and identify the microorganisms without interference from broad-spectrum biocide residues.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the chemical parameters of the treatment solution by using enzymes with specific substrate specificities rather than broad-spectrum biocides. This parameter change allows the treatment to be effective against diverse microorganisms while maintaining compatibility with downstream analytical tests through controlled degradation of only matrix components.

Inventive Principle:
Principle #35Parameter changes

3Reliability

If strong cleaning agents are used to remove biofilms, then biofilm removal is improved, but sensitive bacteria like Listeria are damaged or killed, affecting detection accuracy

Engineering Contradiction:
Improvebiofilm removal effectivenessVSAvoidbacterial sensitivity damage
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention applies local quality by using enzymes that specifically target the extracellular matrix components rather than causing general bacterial damage. Each enzyme acts locally on its specific substrate (proteins, polysaccharides, lipids, cellulose, mannans) within the matrix, degrading it while leaving the bacterial cells themselves intact and viable for accurate detection.

Inventive Principle:
Principle #3Local quality

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The composition effectively collects and preserves microorganisms, including biofilms, for precise identification and targeted treatment, reducing downtime and resource waste by ensuring accurate microbiological analysis.

Implementation Method 1

one or more surfactants, present at a content of at least 0.01% by weight and at most 1.5% by weight relative to the weight of said composition (sum of the weights of surfactants: weight of the composition), said surfactant(s) comprising (or consisting essentially of) an anionic surfactant

Methodology Applied
Scientific EffectSurfactant action: Surfactant

Implementation Method 2

at least two enzymes chosen from the group consisting of a (serine) protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Enzyme

Implementation Method 3

at least two enzymes chosen from the group consisting of a (serine) protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 4

at least one stabilising agent present at a content of at least 15% relative to the weight of said composition, said stabiliser being chosen from the group of polyols (preferably glycerol)

Methodology Applied
Scientific EffectStabilization:

Implementation Method 5

and, preferably at least one sequestering agent

Methodology Applied
Scientific EffectSequestration:

Data Source

PatentUS20250354193A1Composition for removing and collecting microorganisms
Publication Date: 2025.11.20 REALCO SA

AI summary

A sterile sampling composition includes a surfactant, one or more enzymes chosen from the group consisting of a protease, a polysaccharidase, a laccase, a lipase, a cellulase and a mannanase. A sampling kit includes the sterile sampling composition. A sampling method is based on the sterile sampling composition or kit and use of this composition or kit for the detection of bacteria on a surface.