Table 1.

Schematic description of selected standard methods for antimicrobial susceptibility of surfaces.

Standard Method, Title	Test Organisms		Notes
	Bacteria	Fungi
Agar Diffusion Methods
AATCC Test Method 147-2004 Antibacterial Activity Assessment of Textile Materials: Parallel Streak Method	Staphylococcus aureus, Klebsiella pneumoniae		Measurement of inhibition zone after incubation in agar culture in the presence of the test specimen. Designed for diffusible antimicrobial agents.
AATCC Test Method 30-2004 Antifungal Activity, Assessment on textile Materials: Mildew and Rot Resistance of Textile Materials		Aspergillus niger; Penicillium varians; Trichoderma viride	Evaluation using a microscope (50×) of the extent of fungal growth on the test specimens (discs).
BS EN 1104-2018 Paper and board intended to come into contact with foodstuffs-Determination of the transfer of antimicrobial constituents	Bacillus subtilis	Aspergillus niger	Evaluation of the diameter of inhibition zone around each specimen disc.
BS EN ISO 20645/AATCC 90 Textile fabrics, determination of antibacterial activity—Agar diffusion test plate	Staphylococcus aureus, Escherichia coli Klebsiella pneumoniae		Measurement of the inhibition zone after incubation in agar culture in the presence of the test specimen (< or >1 mm).
BS ISO 16869 Plastics assessment of the effectiveness of fungistatic compounds in plastic formulations		Aspergillus niger Chaetomium globosum Penicillium funiculosum Trichoderma longibrachiatum	Observation of fungal growth after pouring the inoculated molten nutrient–salt agar onto the surface of the base agar and test specimen to form a thin second layer.
ASTM E2722-14 Standard test method for using seeded-agar for the screening assessment of antimicrobial activity in fabric and air filter media	Staphylococcus aureus Serratia marcescens	Aspergillus brasiliensis	Designed to qualitatively evaluate the presence of antibacterial and antifungal activity in (or on) fabrics, or air filter media, simulating actual use conditions.
E2922-15 Standard guide for the use of standard test methods and practices for evaluating antibacterial activity on textiles	Staphylococcus aureus, Escherichia coli Klebsiella pneumoniae	Candida albicans	An index of procedures.
Suspension Methods
E2149-20 Standard methods for determining the antimicrobial activity of antimicrobial agents under dynamic contact conditions (fabric, paper, powder and granular material, surfaces)	Escherichia coli		Enumeration of CFU after contact between pieces of specimen with known bacterial concentration on the wrist-action shaker. Designed for non-diffusible antimicrobial agents.
AATCC test method 100-2004 antibacterial finishes on textile materials: assessment of	Staphylococcus aureus, Klebsiella pneumoniae		Designed to measure the antimicrobial activity of textiles after direct inoculation of the textile surface.
JIS L 1902 Testing antibacterial activity and efficacy on textile products	Staphylococcus aureus, Klebsiella pneumoniae		Final microbial concentrations are determined after test fabrics are allowed to incubate undisturbed in sealed containers at body-temperature for 18 h.
ISO 20473-2013 Textile determination of antibacterial activity of textile products	Staphylococcus aureus, Klebsiella pneumoniae		Three inoculation methods are proposed: (a) Absorption method (an evaluation method in which the test bacterial suspension is inoculated directly onto specimens); (b) Transfer method (test bacteria are placed on an agar plate and transferred onto specimens); (c) Printing method (test bacteria are placed on a filter and printed onto specimens).
Quantitative Methods Based on Direct Contact of Microorganisms with Surfaces
JIS Z 2801-2000 Antimicrobial product test for antimicrobial activity and efficacy for plastics	Escherichia coli Staphylococcus aureus,		Enumeration of CFU (after serial dilutions) in the test inoculum applied to the test surface.
ISO 22196/JIS Z 2801 Plastic measurement of antibacterial activity on plastic surfaces	Escherichia coli Staphylococcus aureus		Enumeration of CFU (after serial dilutions) in the test inoculum applied to the test surface.
ASTM E2180-18 Standard test method for determining the activity of incorporated antimicrobial agent(s) in polymeric or hydrophobic materials	Staphylococcus aureus Klebsiella pneumoniae, Pseudomonas aeruginosa		Designed to measure the antimicrobial activity of highly hydrophobic surfaces. The vehicle for the inoculum is an agar slurry, which reduces the surface tension of the saline inoculum carrier and allows formation of a “pseudo-biofilm”, providing more uniform contact of the inoculum with the test surface. The method can confirm the presence of antimicrobial activity in plastics or hydrophobic surfaces and allows determination of quantitative differences in antimicrobial activity between untreated plastics or polymers and those with bound or incorporated low water-soluble antimicrobial agents.
Methods Based on the Biofilm Formation
E3151-18 Standard test method for determining antimicrobial activity and biofilm resistance properties of tube, yarn or fiber specimens	Staphylococcus epidermidis		Determination of planktonic bacteria and adherent bacteria populations that survived exposure to test specimens. Designed for plastic surfaces and other non-porous surfaces including coated substrates