ANTI-MICROBIAL PROPERTIES
From Pathogen Elimination to Barrier Protection
Prebiotics, postbiotics, probiotics, plant extracts, and other functional ingredients are well recognized for their antimicrobial potential, offering promising solutions to support the fight against pathogens.
At ACARYON
We offer a comprehensive portfolio of in vitro screening and analytical assays designed to identify and characterize the antimicrobial properties of your products.
Our expertise spans a wide range of bacterial pathogens, toxins, and viruses, providing deep insights into modes of action and application potential.
In addition to human intestinal cell line models, we employ gastric and vaginal epithelial models, as well as animal intestinal epithelial cell models (porcine, feline, canine and equine models).
Have a non-binding consultation with one of our experts to find out the best solution for your needs.
ANTI-ADHESION PROPERTIES
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Microbial adhesion to host tissues is the critical first step in the establishment of infection, enabling pathogens to anchor, colonize, and subsequently invade. Products with anti-infective properties can interfere with this process either directly by blocking or preventing microbial adhesion or indirectly by modifying host cell surface receptors to reduce pathogen binding or by inducing the production of antimicrobial substances by gastric epithelial cells. By targeting this earliest stage, such interventions can effectively prevent infection before it can develop.
Cell growth and differentiation
(intestinal, gastric, vaginal etc)
Add Test Product
Add
pathogen (labelled
or not)
Pathogen selective agar
Fluorescence quantification
Software-assisted CFU determination
Bioinformatic
& customized visualization
THE ASSAY
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The anti-adhesion assay serves as a valuable first-line screening tool for assessing antimicrobial properties. It reflects a product’s ability to prevent pathogens from binding to host epithelial cells, regardless of the underlying mechanism.
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Whether through direct antimicrobial activity, competitive exclusion, receptor blocking, or pathogen aggregation.
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Follow-up assays, can then be applied to further elucidate the mode of action.
Example of available cells:
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Human intestinal cells line (Caco-2, HT29, HT29-MTX, T84)
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Human Gastric cell line (MKN-45),
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Human vaginal cell line (VK2/E6E7)
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Human skin cells (HaCaT, sebocytes SZ95, Dermal fibroblasts NHDF)
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Porcine intestinal cell line (IPEC-J2)
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Feline intestinal cell line (immortalized intestinal cells)
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Primary cells (intestinal canine cells, etc.)
THE ASSAY
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The Aggregation Assay evaluates the ability of a test compound to induce clumping (aggregation) of pathogenic microorganisms.
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In this assay, the test product is mixed with a suspension of the target pathogen, and the degree of aggregation is monitored visually and quantified spectrophotometrically over time.
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This assay provides valuable insight into the product’s ability to bind and immobilize pathogens, thereby reducing their free and potentially infectious forms
AGGREGATION PROPERTIES
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Microbial aggregation induced by certain products can help limit infection by clumping pathogens together, reducing their ability to adhere to host tissues and initiate colonization. By promoting the formation of large, non-adherent aggregates, these products can also facilitate pathogen clearance, either through natural host defenses or external removal. As a result, aggregation represents a complementary mechanism for preventing or resolving infections.
Bioinformatic
& customized visualization
Aggregates
Pro, Postbiotic strain, ingredient etc.
Pathogen
No Aggregates
CLASSIC ANTI-MICROBIAL PROPERTIES
Classical anti-pathogenic properties of products may include the production of bacteriocins, modification of the microenvironment, or alteration of microbial protein expression (e.g., reduced the expression/secretion of virulence factors)
Depending or the nature of the product and the targeted outcome, the followign assays may be selected:
Overlay assay
Disk/well diffusion assay
Co-culture assay
AMP-production assay
THE OVERLAY ASSAY
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The Overlay Assay is a robust and versatile method for evaluating the antimicrobial activity of probiotic strains, independent of their specific mechanisms of action.
Probiotic strain
Pathogen culture
Overlay
Software assisted determination of Ratio colony/ inhibition Zone
Bioinformatic
& customized visualization
THE ASSAY
In this assay, probiotic colonies are first generated on an agar plate. The plate is overlaid with a soft agar layer containing the target pathogen, enabling direct interaction between the live probiotic cells and the pathogen.
Antimicrobial activity is visualized as inhibition zones surrounding the probiotic colonies, and the antimicrobial potential is quantified by calculating the ratio of inhibition zone diameter to colony diameter, providing a standardized and comparable measure of efficacy.
THE ASSAY
In this assay , a suspension of the target pathogen is uniformly spread onto an agar plate, and the test substance (i.e. supernatant of probiotci culture) is applied either to sterile paper disks placed on the surface or directly into agar wells.
Antimicrobial activity is visualized as clear inhibition zones, and the antimicrobial potential is quantified by calculating the ratio of inhibition zone diameter to well or disk diameter, providing a standardized and comparable measure of efficacy
DISK/WELL DIFFUSION ASSAY
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The disk or well diffusion assay is a widely used method to assess the antimicrobial activity of a substance. This assay evaluates both classical antimicrobial activity (e.g., bacteriocin production) and indirect antagonistic effects of substances (supernatant of probiotic cultures etc.).
Pathogen culture
Cell free supernatans
Software assisted
inhibition Zone measurement
Bioinformatic
& customized visualization
CO-CULTURE ASSAY
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The co-culture assay is used to assess how a test product influences the growth dynamics of pathogens. It allows evaluation of inhibitory versus competitive or lethal effects by monitoring changes in microbial abundance over time, offering a clear readout of antagonistic activity.
Co-culture
Bioinformatic
& customized visualization
Pathogen
Probiotic culture
Ingredient etc
Pathogen selective agar
Growth
kinetics
Software-assisted CFU determination
THE ASSAY
In this assay, a standardized pathogen culture is exposed to the test product under controlled conditions.
Samples are collected at defined time points, and pathogen growth is quantified by spectrophotometric analysis and determination of colony-forming unit (CFU).
AMP PRODUCTION ASSAY
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Antimicrobial peptides (AMPs) are key components of the innate immune defense and are produced by epithelial cells. They can inhibit or kill bacteria, fungi or viruses through diverse mechanisms, including disruption of microbial membranes or interference with essential cellular processes. The AMP production assay provides insight into the ability of test products to stimulate epithelial cells to secrete AMPs.
THE ASSAY
In this assay, intestinal epithelial cells are incubated with the test product under controlled conditions.
After incubation, the cell culture supernatant is collected, and the levels of specific AMPs (defensins, cathelicidins (e.g., LL-37), or REG3γ) are quantified using enzyme-linked immunosorbent assay (ELISA).
Quantification of AMPs
Cell growth and differentiation
Add Test Product
Harvest supernatants
Cell lines and primary cells
-Intestinal cells
-Gastric cells
-Vaginal cells etc
Bioinformatic
& customized visualization
ANTI-CYTOTOXICITY ASSAY
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The main virulence factors of many pathogens are the toxins they produce, which exert cytotoxic effects on host cells and drive the development of disease. By damaging or disrupting cellular functions, these toxins play a central role in pathogenicity. Therefore, preventing or neutralizing toxin activity represents an effective strategy for limiting disease progression and protecting host tissues.
Test Product incubated with toxin
Pre-incubation or direct on cells
(intestinal, gastric, vaginal etc
Fluorescence based viability quantification
Morphology analysis
Available Toxins:
-Cholera toxin
-Heat-labile Toxin
-Shiga Toxin
-C. difficile toxins
-etc
Bioinformatic
& customized visualization
THE ASSAY
Probiotics, postbiotics, and functional ingredients may exert antimicrobial effects by preventing toxin binding to epithelial cells (intestinal, gastric, vaginal, etc.), either through direct interaction with the toxin or by modulating host cell surface properties.
In this assay, the ability of the test product to prevent toxin-induced cytotoxicity and/or inhibit cytokine binding is evaluated using morphological analysis and/or quantitative assessment of cell viability or toxin binding.
Multiple assay variants are available to accommodate specific research objectives.
VIRUS BINDING ASSAY
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Viruses are responsible for a wide range of infections and rely on binding to specific receptors on host cells as the essential first step of infection.
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By interfering with this attachment process, functional products can help block viral entry and thereby reduce or prevent infection.
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Targeting viral binding offers an effective and early intervention strategy to limit viral spread.
In this assay, the test product is incubated with labeled Virus-Like Particles (VLPs). After incubation, unbound VLPs are removed by separating the supernatant.
Bound VLPs are then detected using proprietary ELISA assays.
Complementary cell-based assays for virus-specific anti-adhesion are also available. Please contact us for customized solutions tailored to your research needs.
THE ASSAY
Labeled VLPs
Incubation test Product with Labeled-VLPs
Common viruses (VLPs) of interest:
-Rotavirus
-Norovirus
-Astrovirus
-Influenza
-Coronavirus
-HPV etc.
Bioinformatic
& customized visualization
Quantification of adhesion
Have a non-binding consultation with one of our experts to find out the best solution for your needs.
Contact us.
