E35 - Pesticides, Antimicrobials, and Alternative Control Agents

SIGNIFICANCE AND USE
5.1 An ideal wound dressing absorbs wound exudates, reduces the bioburden within/on the dressing, and protects wounds from microbial penetration through the dressing. Microbial barrier effectiveness testing is performed to measure the ability of microorganisms to penetrate through a material under specific conditions.  
5.2 The test may be used for solid wound dressings with or without the addition of antimicrobial agents where the manufacturer is claiming microbial barrier properties in their performance claim. Barriers can be physical (polyurethane) and/or chemical (antimicrobial barrier).  
5.2.1 Five (5) commonly used solid wound dressings are (1) hydrogel, (2) hydrocolloid, (3) film, (4) foam, and (5) alginate. Liquid adhesives also protect the wound from being infected by microorganisms. Liquid adhesive dressings contain cyanoacrylate or polyacrylate, which polymerizes quickly to create a solid barrier and protect the wound from contamination.  
5.3 This qualitative test method can be used to determine the microbial barrier properties of solid wound dressings including liquid adhesive.  
5.3.1 The test represents an in vitro wound model by using a nutrient-rich medium to mimic the wound, the contact time will represent the maximum period of use for a single dressing application on the wound, and an incubation temperature of 37 °C ± 2 °C represents the human body temperature.  
5.4 Results are documented as “Growth” or “No Growth.”  
5.4.1 No growth underneath the dressing indicates the barrier properties of the wound dressing are effective at preventing microbial penetration through the dressing.  
5.4.2 Growth underneath the dressing demonstrates that the challenge organism was able to penetrate the dressing and reach the media surface, indicating that microorganisms can reach the wound by penetrating through the dressing from the outside environment.
SCOPE
1.1 This test method is used to determine the microbial barrier properties of wound dressing. The test is designed to measure the ability of microorganisms to penetrate through a material (wound dressing) under specific conditions. This qualitative in vitro test will demonstrate results such as “Growth” or “No Growth” of the test organism on the agar surface after a specified contact time.  
1.2 The test method includes an initial inoculum of 3.0 × 106 ± 2.0 × 106 CFU/mL and a contact time of 24 h to 72 h or per the wound dressing label claim. To show barrier properties, wound dressing should not allow the challenged organism to penetrate and reach the media surface.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 Testing is to be performed by individuals trained in microbiological techniques under appropriately controlled conditions to ensure the integrity of results and personnel safety.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This method can be used to evaluate effectiveness of incorporated/bound antimicrobials in hydrophobic materials such as plastics, epoxy resins, as well as other hard surfaces.  
5.2 The aqueous based bacterial inoculum remains in close, uniform contact in a “pseudo-biofilm” state with the treated material. The percent reduction in the surviving populations of challenge bacterial cells at 24 h versus those recovered from a non-treated control is determined.  
5.3 The hydrophobic substrate may be repeatedly tested over time for assessment of persistent antimicrobial activity.
SCOPE
1.1 This test method is designed to evaluate (quantitatively) the antimicrobial effectiveness of agents incorporated or bound into or onto mainly flat (two dimensional) hydrophobic or polymeric surfaces. The method focuses primarily on assessing antibacterial activity; however, other microorganisms such as yeast and fungal conidia may be tested using this method.  
1.2 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 even contact of the inoculum with the test surface.
Note 1: This test method facilitates the testing of hydrophobic surfaces by utilizing cells held in an agar slurry matrix. This test method, as written, is inappropriate to determine efficacy against biofilm cells, which are different both genetically and metabolically than planktonic cells used in this test.  
1.3 This 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. Comparisons between the numbers of survivors on preservative-treated and control hydrophobic surfaces may also be made.  
1.4 The procedure also permits determination of “shelf-life” or long term durability of an antimicrobial treatment which may be achieved through testing both non-washed and washed samples over a time span.  
1.5 Knowledge of microbiological techniques is required for these procedures.  
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The plastic Petri plate (carrier) provides a closed system for enumeration and easy application of a pre-saturated or impregnated antimicrobial towelette by an analyst.  
5.2 Aliquoting of sterile 5 % non-heat-inactivated fetal bovine serum (five 10 µL spots) onto soiled carriers and inoculation of final test suspension onto treated carriers (five 10 µL spots) is conducted using a template and a positive displacement pipette, thereby ensuring a precise inoculum level and uniform distribution of soil and final test suspension.  
5.3 A single towelette is tested per 2-carrier set, eliminating the likelihood of cross contamination between carriers.  
5.4 The corkscrew-patterned circular motion of the product application (wipe outside to inside, wipe inside to outside using the wiping template; see Annex A3 – Annex A6) ensures uniform coverage and contact of disinfectant with the inoculated surface.  
5.5 The addition of neutralizer to the treated carriers at the end of the contact time results in neutralization of the test substance. This standard test method provides a procedure for performing neutralization verification to confirm that the microbicidal, microbistatic, or both types of activity of a test substance has been reduced by 50 % at the end of the contact time (see Annex A1 for neutralization verification procedure).  
5.6 The design of this standard test method minimizes any loss of viable organisms through carrier wash-off.  
5.7 It is optional to adjust (dilution in PBS) the inoculum to achieve desired control counts of 5.0 log10 CFU/carrier to 6.5 log10 CFU/carrier.  
5.8 Include, where applicable, comparisons of the test to other similar procedures such as Practices E1054 and E2362.
SCOPE
1.1 This test method quantitatively determines the effectiveness of various sizes of antimicrobial towelettes in treating hard, non-porous surfaces against Pseudomonas aeruginosa and Staphylococcus aureus.  
1.2 This test method may be used to evaluate towelettes for antimicrobial efficacy against additional microorganisms (with necessary modifications).  
1.2.1 This test method does not differentiate between chemical inactivation of the test microbe and mechanical removal of inoculum from a surface; rather, product efficacy is considered a combination of both attributes of a towelette-based formulation.  
1.3 This test method involves the use of hazardous materials, chemicals, and infectious microorganisms and therefore should be performed only by those trained in microbiological techniques in facilities designed and equipped for work with infectious agents at the appropriate biosafety level, a BSL-2 or higher laboratory; specifications provided in the “Biosafety for Biomedical and Microbiological Laboratories” (BMBL), 6th edition (BMBL).  
1.4 It is the responsibility of the investigator to determine whether Good Laboratory Practices (GLP Standards—For example, 40 CFR, Part 160 of FIFRA) are required and to follow them when appropriate.  
1.5 Strict adherence to the protocol is necessary for the validity of the test results.  
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 Extensional viscosity is a measure of the resistance of a liquid to stretching forces, such as those occurring during the disruption of liquid films and the formation of sprays used in agriculture and other purposes including painting operations or metal working. This method for measurement of a Screen Factor, gives a relative value for extensional viscosity, which may be used:  
4.1.1 To compare the potential for drift control of different polymers.  
4.1.2 To compare the relative extensional viscosity component of different spray tank mixtures.  
4.1.3 To determine the extent of breakdown of polymer solutions used as drift control additives during the recirculation of the solutions through pumps and screens.  
4.1.4 To use as a parameter in the Spray Drift Task Force Models for droplet size prediction.  
4.2 It should also be noted that many drift control polymers are irreversibly destroyed during the recirculation of spray mixes by pumping with high shear pumps such as gear or centrifugal pumps. It is advisable to subject the test mixture to similar pumping regimes to simulate practical conditions before carrying out the extensional viscosity test. Measurements of extensional viscosity are the only presently known method of determining the extent of this breakdown properties of dilute polymer solutions.  
4.3 This method is intended to produce a relative value for extensional viscosity. The purpose of the method is to compare the extensional viscosity produced by different polymer types or concentrations of polymer in spray tank mixes.
SCOPE
1.1 This test method covers the determination of the relative extensional viscosity or Screen Factor (SF) of dilute agricultural spray mixes.  
1.2 The test can be used for tank mixes containing dissolved, emulsified or dispersed materials, or mixtures.  
1.3 Results may be affected by the quality of the water used. Make-up water quality should therefore be specified in the presentation of results.  
1.4 Proper safety and hygiene precautions must be taken when working with pesticide formulations to prevent skin or eye contact, vapor inhalation, and environmental contamination. Read and follow all handling instructions for the specific formulation and conduct the test in accordance with good laboratory practice.
Note 1: References to the development of extensional viscosity from dissolved polymers, extensional viscosity effects on the droplet size distribution of sprays, and measurements of screen factor on recirculated spray mixes containing polymers are available.2,3  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Guide D6469 details the types of problems associated with uncontrolled microbial growth in fuels and fuel systems. Treatment with effective antimicrobial agents is one element of contamination control strategy.  
5.2 The procedure should be used to evaluate the relative efficacy of microbicides in liquid fuels boiling below 390 °C. The effect of environmental conditions, such as a variety of fuel additives, metal surfaces, and climatology, are variables that can be included in specific tests using this protocol.  
5.3 This practice addresses product performance issues only. Regulatory Agencies restrict and control the use of both pesticides (in the U.S.: 40 CFR 152) and fuel additives (40 CFR 79). Regardless of performance in this method, antimicrobials must only be used in compliance with applicable regulations. Specific industries, for example, the aviation industry, may place further restrictions on chemicals used for fuel treatment.
SCOPE
1.1 This practice is designed to evaluate antimicrobial agents for the prevention of microbially influenced deterioration of liquid fuels (as defined by Specification D396, D910, D975, D1655, D2069, D2880, D3699, D4814, D6227, D6751, and D7467), system deterioration, or both.  
1.2 Knowledge of microbiological techniques is required for these procedures.  
1.3 It is the responsibility of the investigator to determine whether Good Laboratory Practice (GLP) is required and to follow them where appropriate (40 CFR, 160), or as revised.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This test method provides for rapid screening of antimicrobial treatments located in or on the carpet face fiber or incorporated into the backing structure of the carpet (or both).  
5.2 This test method simulates actual use conditions that may occur on carpets (for example, food and beverage spills, soiling from foot traffic, prolonged moisture exposure).  
5.3 This test method provides a means to screen for activity and durability of an antimicrobial treatment under conditions of organic loading.  
5.4 This test method provides for the simultaneous assessment of multiple carpet components for antimicrobial activity.  
5.5 Carpets may be cleaned prior to testing with this test method in order to assess the durability of the antimicrobial effect.
SCOPE
1.1 This test method is designed to evaluate (qualitatively) the presence of antimicrobial activity in or on carpets. Use this test method to qualitatively evaluate both antibacterial and antifungal activity.  
1.2 Use half strength (nutrient and agar) tryptic soy agar as the inoculum vehicle for bacteria and half strength potato dextrose agar as the inoculum vehicle for mold conidia. Use of half strength agars may reduce undue neutralization of an antimicrobial due to excessive organic load.  
1.3 This test method simultaneously evaluates (both visual and stereo-microscopic) antimicrobial activity both at the fiber layer and at the primary backing layer of carpet.  
1.4 Use this test method to assess the durability of the antimicrobial treatments on new carpets, and on those repeatedly shampooed or exposed to in-use conditions.  
1.5 Knowledge of microbiological techniques is required for the practice of this test method.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This practice is to be used for the removal of virucidal agents from test product-virus mixtures, or from test product-neutralizer-virus mixtures, at or after the contact period and before the inoculation of these mixtures into host systems for assay of viral infectivity.  
5.2 The purpose of the practice is to reduce the concentration of the cytotoxic properties of the test product and neutralizers in order to permit the evaluation of viral infectivity at dilutions that would otherwise be toxic to the host cells.  
5.3 The practice is applicable to the testing of liquid, pre-saturated towelettes, and pressurized disinfectant products, as well as handwash/rub products.  
Note 3: When testing products, the ability of the solution to pass through the column must be verified prior to testing. Certain products with high viscosities are unable to pass through columns. If the product is determined to be too viscous, alternative neutralization methods should be employed.  
5.4 This practice is compatible with organic soil loads, hard water, disinfectants containing organic solvents, and chemical neutralizers.
SCOPE
1.1 This practice is intended to be used to reduce the cytotoxic level of the virus-test product mixture prior to assaying for viral infectivity. It is used in conjunction with evaluations of the virucidal efficacy of disinfectant solutions, wipes, trigger sprays, or pressurized disinfectant spray products intended for use on inanimate, nonporous environmental surfaces. This practice may also be used in the evaluation of hygienic handwashes/handrubs, or for other special applications. The practice may be employed with all viruses and host systems.
Note 1: Gel filtration columns may impact virus titer and their use should be taken into consideration when selected for use.  
1.2 This practice should be performed only by persons trained in virology techniques.  
1.3 This practice utilizes gel filtration technology. The effectiveness of the practice is dependent on the ratio of gel bed volume to sample size and uniformity in the preparation of columns as well as the conditions of centrifugation. The effectiveness of this practice is maximized by investigator practice and experience with gel filtration techniques.  
1.4 This practice will aid in the reduction, but not necessarily elimination, of test product toxicity while preserving the titer of the input virus.  
1.5 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method was designed principally for clay granular carriers and clay-based granular formulations, but need not be limited to these materials.  
4.2 This procedure is applicable to granules in the range from 6 to 80 mesh (3.36 to 0.17 mm).  
4.3 The sieve sizes used to calculate total particle count will be called the desired range and should be specified as part of the test results.
SCOPE
1.1 This test method is used to determine the number of particles per pound of granular carriers and granular pesticide formulations.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific precautionary statements, see Section 6.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 Fungi are known to produce objectionable odors, stains, and premature biodeterioration of various consumer products and construction substrates including textiles, carpet, ceiling tile, gypsum wallboard, lumber, and plasticized vinyl and other polymers.  
4.2 Antifungal activity is typically:  
4.2.1 Determination of article susceptibility to fungal colonization,  
4.2.2 Determination of fungistatic activity (qualitative determination of prevented or delayed fungal colonization), and  
4.2.3 Determination of fungicidal/sporicidal activity (quantitative determination of spore kill).  
4.3 The degree of required surface examination varies from gross visual examination to detailed microscopic assessment among these methods.  
4.4 This guide provides an overview of established methods and suggestions for their applicability, with consideration to the type of substrate treated or the type of antifungal treatment being assessed.
SCOPE
1.1 This guide provides information on various test methods currently available to assess antifungal activity on natural or synthetic substrates.  
1.2 Knowledge of microbiological techniques is required for the practice of this guide.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 Antimicrobial agents are routinely used for treating textile materials for the reduction of biodeterioration and bacterial odor generation. Furthermore, textiles are treated to prevent or limit microbial cross-contamination in healthcare settings.  
4.2 Antimicrobial agents used in textiles will vary with regard to their broad-spectrum effectiveness, biostatic/biocidal properties, and binding properties in or on particular substrates. When selecting antibacterial test methods as the sole means to predict end use behavior it is critical to understand the intended end use conditions of the treated articles.  
4.3 Textile materials differ with regard to the knit/weave, fabric composition, and added functional feature (for example, water repellent, flame retardant, softener, whitener). Each of these factors may alter test results within a given method.  
4.4 The test methods indicated below differ mainly in the procedure for inoculating samples, levels of nutrients in the bacterial challenge, organisms used, exposure times, and procedure for sterilization of test samples. Each of these parameters are often subject to industry modifications.  
4.5 Some antimicrobial treated articles are not suitable for sterilization due to the sensitivity of these antimicrobial agents to high temperature and humidity. Furthermore, some antimicrobial agents may be unrealistically activated due to UV sterilization which could show false positive antimicrobial properties. Sterilization of test fabrics prior to testing should be avoided if possible. All modifications of the methods indicated below should be clearly indicated on associated test reports and should be appropriate to the antimicrobial technology used.  
4.6 This guide is intended to review each commonly used industry test standard for its applicability with an understanding of each of the factors listed above. Further, it is the intention of this guide to indicate commonly used and generally accepted modifications of e...
SCOPE
1.1 This guide provides users with an index of procedures in the form of test methods, practices, and related international documents that are currently used in the textile industry for determining antibacterial properties of antimicrobial treated textile articles. This guide is not considered as all-inclusive for antimicrobial testing procedures related to textiles.  
1.2 This guide identifies some existing ASTM and other industry standard test methods applicable for testing the antibacterial performance on textiles and discusses options within each method that have been used to address specific end-use performance expectations in addition to measuring wash durability of such activity.  
1.3 This guide is intended to assist testing facilities in determining which test methods are appropriate for which treated articles based on type of antimicrobial active involved (diffusible versus non-diffusible), nature of test fabric, and expected end use.  
1.4 The test methods indicated in this guide should be performed only by those trained in microbiological techniques, are familiar with textile antimicrobial agents and with the end use exposures of the antimicrobial treated textile material.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 These procedures should be used to test topical antimicrobial-containing preparations that are intended to be fast-acting in reducing significantly the number of microorganisms on intact skin immediately and, for preoperative and vascular precatheterization preparations, maintenance of some degree of reduction for an extended time.
SCOPE
1.1 The practice is designed to measure the reduction of the microflora of the skin.  
1.2 A knowledge of microbiological techniques is required for these procedures.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3.1 Exception—In this practice, metric units are used for all applications except for linear measure, in which case inches are used, and metric units follow in parentheses.  
1.4 Performance of this procedure requires a knowledge of regulations pertaining to the protection of human subjects (1).2
Note 1: Importantly, it must be noted that the FDA currently does not accept data resulting from procedures in this Method for testing products for approval as Vascular Precatheterization Skin Preparations.  
1.5 This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Bacteria that exist in a biofilm are phenotypically different from suspended cells of the same genotype. The study of biofilm in the laboratory requires protocols that account for this difference. Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. The purpose of this method is to direct a user in the laboratory study of biofilms by clearly defining each system parameter. This method will enable a person to grow, sample, and analyze a laboratory biofilm. The method was originally developed to study toilet bowl biofilms, but may also be utilized for research that requires a biofilm grown under moderate fluid shear.
SCOPE
1.1 This test method is used for growing a reproducible (1)2 Pseudomonas aeruginosa biofilm in a continuously stirred tank reactor (CSTR) under medium shear conditions. In addition, the test method describes how to sample and analyze biofilm for viable cells.  
1.2 Although this test method was created to mimic conditions within a toilet bowl, it can be adapted for the growth and characterization of varying species of biofilm (rotating disk reactor—repeatability and relevance (2)).  
1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log10 colony forming units per surface area (rotating disk reactor—efficacy test method (3)).  
1.4 Basic microbiology training is required to perform this test method.  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This procedure may be used to assess the in vitro reduction of a microbial population of test organisms after exposure to a test material.
SCOPE
1.1 This guide covers an example of a method that measures the changes in a population of aerobic microorganisms within a specified sampling time when antimicrobial test materials are present.  
1.1.1 Several options for organism selection and growth, inoculum preparation, sampling times and temperatures are provided.  
1.1.2 When the technique is performed as a specific test method, it is critical that the above mentioned variables have been standardized.  
1.1.3 Antimicrobial activity of specific materials, as measured by this technique, can vary significantly depending on variables selected.  
1.1.4 Test Method E2783 may be referenced as an example of using fixed conditions and set variables to evaluate antimicrobial efficacy of water-miscible compounds.  
1.1.5 This guide serves as a general teaching document for evaluating the antimicrobial activity using a variety of conditions to offer the flexibility needed in test conditions to cover a broad range of microorganisms and test substances.  
1.1.6 It is important to understand the limitations of in vitro tests, especially comparisons of results from tests performed with different parameters. As an example, test results of microorganisms requiring growth supplements or special incubation conditions may not be directly comparable to organisms evaluated without those stated conditions.  
1.2 Knowledge of microbiological techniques is required for this procedure.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This in vivo procedure is designed to test the ability of hygienic handwash or handrub agents to eliminate fungal contamination from experimentally-contaminated hands. Since the two thumbpads and all eight fingerpads can be used in any given test, it allows for the incorporation of an input control (two), control for culturable cells of the test fungus remaining after the inoculum has dried (two), fungal cells eliminated after treatment with a control or reference solution (two), and up to four replicates to assess the fungus-eliminating efficiency of the formulation under test. No more than 100 µL of the test fungal suspension is required to complete one test.  
5.2 Whereas this practice is designed to work with fungi, similar ASTM standards exist for testing against viruses (Test Method E1838) and vegetative bacteria (Test Method E2276).  
5.3 The levels of culturable microorganisms left on hands after washing can be reduced further by drying the washed hands with paper, cloth, or warm air (5). A step for the drying of fingerpads after exposure to the control or test solution, therefore, has not been included to avoid fungal removal by the drying process itself.  
5.4 This practice is not designed to test surgical hand scrubs or preoperative skin preps.  
5.5 The level of contamination with culturable fungi on each fingerpad after the drying of the inoculum should be at least 104 CFU so that it would permit the detection of up to a 4-log10 reduction in the viability titer of the test organism by a test formulation under the conditions of this test. This in itself does not represent the product performance criterion, which may vary depending on the jurisdiction and the nature of the formulation being evaluated.
SCOPE
1.1 This practice is designed to assess the ability of hygienic handwash and handrub agents to reduce levels of fungal contamination on hands (3) . This practice is not meant for use with surgical hand scrubs (Test Method E1115) or preoperative skin preps (Test Method E1173).  
1.2 Performance of this procedure requires the knowledge of regulations pertaining to human experimentation.3  
1.3 The practice should be performed by persons with training in microbiology in facilities designed and equipped for work with infectious agents at biosafety level 2 (4).  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The procedure can be incorporated into protocols used to evaluate test materials containing antibacterial ingredients that are intended to reduce significantly the number of organisms on intact skin. It also may be used to provide an indication of residual antibacterial activity (as in Guide E2752). Examples of test materials, for which this practice is applicable, include pre-operative skin preparations, hand-washes, surgical scrubs, acne reduction products, and others. For each type of test material, types of resident flora or surrogate organisms, or a combination thereof, may differ and should be considered (this is, aerobic bacteria, anaerobic bacteria, yeast, or mold).  
5.2 The procedure may be used in protocols intended to evaluate and identify resident flora from the skin.  
5.3 Performance of this technique may require the knowledge of regulations pertaining to the protection of human subjects if the protocol involves application of the technique to the skin of human subjects.
SCOPE
1.1 This practice is designed to recover microorganisms from the skin of human subjects or human subject surrogates (animal skin, isolated porcine skin, human skin equivalents, and other such surfaces).  
1.2 Knowledge of microbiological techniques is required for these procedures.  
1.3 It is the responsibility of the investigator to determine if Good Laboratory Practice (GLP) and Good Clinical Practice (GCP) is required.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Methods such as D3273 Standard Test Method for Resistance to Growth of Mold on the Surface of Interior Coatings in an Environmental Chamber and D3274 Standard Test Method for Evaluating the Degree of Surface Disfigurement of Paint Films by Fungal or Algal Growth or Soil or Dirt Accumulation provide means for assessing mold and algal staining on paints. The Test Method E1428 Evaluating the Performance of Antimicrobials in or on Polymeric Solids Against Staining by Streptomyces species (A Pink Stain Organism) is used for solid polymeric materials, but is not appropriate for all antimicrobial technologies.  
5.2 This test method provides a technique for evaluating antimicrobials in or on polymeric materials against staining by Streptomyces species and should assist in the prediction of performance of treated articles under actual field conditions.
SCOPE
1.1 This test method is intended to assess susceptibility of polymer materials, as well as products that may directly contact the treated polymer, to staining by the Actinomycete Streptomyces species.  
1.2 This test method is also suitable for evaluating dark-pigmented test samples since the bacterial growth inhibition can be assessed.  
1.3 Familiarity with microbiological techniques is required. This test method should not be used by persons without at least basic microbiological training.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This method can be used to evaluate the effectiveness of incorporated or bound anti-adherent agents in synthetic polymeric materials and polymeric coatings intended to reduce the attachment of bacteria to the substrate surface.  
5.2 The synthetic polymeric substrate surface may be tested repeatedly over time for assessment of persistent ability of a material to resist bacterial adherence.  
5.3 This method is to quantify the degree of bacteria colonization of a surface to assess a materials ability to resist bacterial adherence because biofilm formation can contribute to material degradation and malfunction.
SCOPE
1.1 This method is designed to evaluate (quantitatively) the number of bacteria attached to the flat, two-dimensional surfaces of synthetic polymeric materials and polymeric coatings on various substrates that may or may not contain bound or incorporated anti-adherent agents. The method focuses on assessing the ability of the surface to reduce bacterial attachment. Other microorganisms such as yeast and fungal conidia may be tested using this method.  
1.2 This test method quantitatively determines the differences in bacterial adherence seen between synthetic polymeric surfaces that allow bacterial adherence and those that do not, comparing the number of organisms recovered from the control surface to the number recovered from the test specimen surface after the contact time. Knowledge of microbiological techniques is required for these procedures.  
1.3 This test method specifies proper methods for measuring the ability of a synthetic polymeric material to resist adherence against specified organism. Due to individual sensitivities, the result of one test organism might not be applicable for other organisms.  
1.4 This test method is designed to measure the potential ability to resist bacterial adherence of a non-porous surface compared directly to a polyester control panel known to support bacterial adherence under specific testing conditions.  
1.5 Antimicrobial treated non-porous surfaces may demonstrate ability to resist bacterial adherence in this method. This method does not purport to differentiate between anti-adherence and antimicrobial activity nor is it designed to reflect specific end-use or environmental conditions. Any product that demonstrates ability to resist bacterial adherence in this method should be measured for antimicrobial activity using a separate test technique such as Test Method E2180 or ISO 22196.  
1.6 The method focuses on assessing the ability of synthetic polymeric materials and polymeric coatings on various substrates to reduce bacterial attachment. The specimen with absorbing or adhesive surfaces may be unable to be disinfected properly before testing, or may trap inoculated organism during recovery process and thus lead to a false result. This method does not apply to specimens with absorbent or adhesive surfaces.  
1.7 The values stated in SI units are to be regarded as standard. The values given in parentheses after SI units are provided for information only and are not considered standard.  
1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method shall be used to determine if a chemical intended for use as a non-food contact sanitizer or as a one-step cleaner-sanitizer provides percent reductions of the selected test organisms on treated carriers as compared to control.
SCOPE
1.1 This test method is used to evaluate the antimicrobial efficacy of sanitizers on precleaned, inanimate, hard, nonporous, non-food contact surfaces against Staphylococcus aureus, or Klebsiella pneumoniae or Klebsiella aerogenes, or a combination thereof. Appropriate modifications to the method may be required when testing organisms not specified herein. When utilizing test surfaces not described herein (see Test Method E2274) or when evaluating spray-based or towelette-based antimicrobial products, modifications may also be required.  
1.2 This test method may also be used to evaluate the antimicrobial efficacy of one-step cleaner-sanitizer formulations recommended for use on lightly soiled, inanimate, nonporous, non-food contact surfaces.  
1.3 It is the responsibility of the investigator to determine whether Good Laboratory Practices (GLP) are required and to follow them where appropriate (see section 40 CFR, 160 or as revised.)  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard may involve hazardous materials, chemicals and microorganisms and should be performed only by persons who have had formal microbiological training.  This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
2.1 This guide provides information on the calibration of boom-type field sprayers used for broadcast, band, or row applications. It is easily adapted to boomless and air-carrier type sprayers for broadcast applications and to rotary atomizers such as spinning disks. A single calibration procedure that is applicable in all situations is not feasible because of the nature of the variables involved in pesticide application. Electronic controllers and other aids should be calibrated by one of these methods.
SCOPE
1.1 This guide is for those who prepare field sprayer calibration procedures. The purpose is to encourage methods that will improve uniformity, accuracy, and safety of pesticide application with field sprayers.  
1.2 The values stated in SI units are to be regarded as standard. The values given in parentheses are mathematical conversions to inch-pound units that are provided for information only and are not considered standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  For specific hazard statements see Section 4.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The effectiveness of antimicrobial agents incorporated into disinfectants, sanitizers, and antiseptics is measured by their ability to kill microorganisms within a specified contact time. Hence, accurate determination of antimicrobial effectiveness requires complete and immediate inactivation (neutralization) of the antimicrobial agent. Inefficient or incomplete neutralization will permit killing or inactivation of microorganisms to continue beyond the experimental exposure time, resulting in an overestimation of antimicrobial activity.  
5.2 The neutralization methods commonly used in antimicrobial effectiveness evaluations are chemical inactivation, dilution, and filtration. All critical parameters of an antimicrobial effectiveness evaluation—for example, media, equipment, microorganism(s), and temperature of solutions—must be duplicated in the performance of selected neutralization procedure.  
5.3 The neutralization evaluation must include at least three replications (five replications in Section 9) so that a statistical analysis of the microbial recovery data can be performed. The number of replicates used in the evaluation depends on the statistical significance required for the expected results, the variability encountered in the data, and the relative effectiveness of the neutralization procedure.  
5.4 A limitation of these evaluation procedures is that they use microorganisms that have not been exposed to an antimicrobial agent. Under experimental conditions, cells exposed to neutralization procedures are likely to be damaged to different degrees by the antimicrobial agent. Sublethal injury may be a factor in recovery, and the effect of the neutralization procedure on recovery of injured organisms should be examined. This method is not intended to assess recovery of injured organisms.
Note 3: Ideally, all microorganisms used in the antimicrobial effectiveness evaluation should be tested in the neutralization assay. However, representative organism...
SCOPE
1.1 These test procedures are used to determine the effectiveness of methodologies procedures and materials intended for inactivating (neutralizing, quenching) the microbicidal properties of antimicrobial agents; to ensure that no components of the neutralizing procedures and materials, themselves, exert an inhibitory effect on microorganisms targeted for recovery; and to demonstrate that the antimicrobial chemistry tested is microbicidal.  
1.2 Knowledge of microbiological and statistical techniques is required for these procedures.
Note 1: These methods are not suitable when testing the virucidal activity of microbicides (see Test Method E1482).  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This practice describes a procedure for producing spore suspensions of C. difficile ATCC 700792, C. difficile ATCC 43598, or C. difficile ATCC 43599. The spore suspensions may be used in antimicrobial efficacy testing, or other laboratory testing requiring C. difficile spores. A spore crop is considered acceptable if the titer is >8 log10 spores/mL, purity of 95 %, and is resistant to 2.5M HCl after 10 min of exposure.
SCOPE
1.1 This practice is designed to propagate spores of Clostridioides difficile using liver broth.  
1.2 It is the responsibility of the user of this practice to determine whether Good Laboratory Practices are required and follow when appropriate.  
1.3 This practice should only be performed by those trained in microbiological techniques.  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This practice may be used to determine if a pre-saturated or impregnated towelette demonstrates antimicrobial effectiveness as a disinfectant on hard surfaces. This practice provides survivor results in the form of a qualitative endpoint (growth positive versus growth negative). The results generated by following this practice do not provide for specific quantitative reductions.
SCOPE
1.1 This practice is designed to evaluate the antimicrobial activity of pre-saturated or impregnated towelettes when used as a hard surface disinfectant.  
1.2 It is the responsibility of the investigator to determine whether Good Laboratory Practices (GLP’s) are required and to follow them when appropriate.  
1.3 This practice should be performed only by those trained in microbiological techniques.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 Appropriate modifications to the practice may be required when testing organisms not specified herein.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilms with specific characteristics. Altering either the engineered system or operating conditions will modify those characteristics. The goal in biofilm research and efficacy testing is to choose the growth reactor that generates the most relevant biofilm for the particular study.  
5.2 The purpose of this test method is to direct a user in how to grow, treat, sample and analyze a Pseudomonas aeruginosa biofilm using the MBEC Assay. Microscopically, the biofilm is sheet-like with few architectural details as seen in Harrison et al (6). The MBEC Assay was originally designed as a rapid and reproducible assay for evaluating biofilm susceptibility to antibiotics (2). The engineering design allows for the simultaneous evaluation of multiple test conditions, making it an efficient method for screening multiple disinfectants or multiple concentrations of the same disinfectant. Additional efficiency is added by including the neutralizer controls within the assay device. The small well volume is advantageous for testing expensive disinfectants, or when only small volumes of the disinfectant are available.
SCOPE
1.1 This test method specifies the operational parameters required to grow and treat a Pseudomonas aeruginosa biofilm in a high throughput screening assay known as the MBEC (trademarked)2 (Minimum Biofilm Eradication Concentration) Physiology and Genetics Assay. The assay device consists of a plastic lid with ninety-six (96) pegs and a corresponding receiver plate with ninety-six (96) individual wells that have a maximum 200 μL working volume. Biofilm is established on the pegs under batch conditions (that is, no flow of nutrients into or out of an individual well) with gentle mixing. The established biofilm is transferred to a new receiver plate for disinfectant efficacy testing.3, 4 The reactor design allows for the simultaneous testing of multiple disinfectants or one disinfectant with multiple concentrations, and replicate samples, making the assay an efficient screening tool.  
1.2 This test method defines the specific operational parameters necessary for growing a Pseudomonas aeruginosa  biofilm, although the device is versatile and has been used for growing, evaluating and/or studying biofilms of different species as seen in Refs (1-4).5  
1.3 Validation of disinfectant neutralization is included as part of the assay.  
1.4 This test method describes how to sample the biofilm and quantify viable cells. Biofilm population density is recorded as log10 colony forming units per surface area. Efficacy is reported as the log10 reduction of viable cells.  
1.5 Basic microbiology training is required to perform this assay.  
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.7 ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentioned in this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the risk of infringement of such rights, are entirely their own responsibility.  
1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.  
1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Bacteria that exist in biofilms are phenotypically different from suspended cells of the same genotype. Research has shown that biofilm bacteria are more difficult to kill than suspended bacteria (5, 7). Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. For example, research has shown that biofilm grown under high shear is more difficult to kill than biofilm grown under low shear (5, 8). The purpose of this test method is to direct a user in the laboratory study of a Pseudomonas aeruginosa biofilm by clearly defining each system parameter. This test method will enable an investigator to grow, sample, and analyze a Pseudomonas aeruginosa biofilm grown under high shear. The biofilm generated in the CDC Biofilm Reactor is also suitable for efficacy testing. After the 48 h growth phase is complete, the user may add the treatment in situ or remove the coupons and treat them individually.
SCOPE
1.1 This test method specifies the operational parameters required to grow a reproducible (1)2 Pseudomonas aeruginosa ATCC 700888 biofilm under high shear. The resulting biofilm is representative of generalized situations where biofilm exists under high shear rather than being representative of one particular environment.  
1.2 This test method uses the Centers for Disease Control and Prevention (CDC) Biofilm Reactor. The CDC Biofilm Reactor is a continuously stirred tank reactor (CSTR) with high wall shear. Although it was originally designed to model a potable water system for the evaluation of Legionella pneumophila (2), the reactor is versatile and may also be used for growing and/or characterizing biofilm of varying species (3-5).  
1.3 This test method describes how to sample and analyze biofilm for viable cells. Biofilm population density is recorded as log10 colony forming units per surface area.  
1.4 Basic microbiology training is required to perform this test method.  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The guide may be used to demonstrate the effectiveness of topical antimicrobial products using pigskin as a surrogate for human skin and the cup scrub technique for sampling.  
5.2 The techniques described can be used to simulate Test Method E1174 and will use the pigskin substrate to overcome limitations posed by exposure of human subjects to potentially pathogenic microorganisms, while offering the benefit of applicability to a wide variety of hand-washing conditions that cannot be simulated in test tubes.  
5.3 Use of the pigskin surrogate offers less expensive and higher throughput screening.
SCOPE
1.1 This guide is designed to demonstrate the effectiveness of hand hygiene topical antimicrobial products using pigskin as a surrogate model.  
1.2 Knowledge of microbiological techniques is required for these procedures.  
1.3 This standard guide can be used to evaluate topical antimicrobial handwash or handrub formulations.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This procedure may be used to assess the in vitro reduction of a microbial population of test organisms after exposure to a test material.
SCOPE
1.1 This test method measures the changes of a population of aerobic and anaerobic microorganisms within a specific sampling time when tested against antimicrobial test materials in vitro. The organisms used are standardized as to growth requirements and inoculum preparation and must grow under the conditions of the test. The primary purpose of this test method is to provide a set of standardized conditions and test organisms to facilitate comparative assessments of antimicrobial materials miscible in aqueous systems.  
1.2 This test method allows the option of using a test sample size of 10 mL or 100 mL.  
1.3 Knowledge of microbiological techniques is required for this procedure.  
1.4 Aseptic technique should be practiced at all times.  
1.5 In this test method, SI units are used for all applications, except for distance in which case inches are used and SI units follow in parentheses.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Hand hygiene is considered one of the most important measures for preventing the spread of infectious microorganisms. Hand rubs reduce the microbial load on the hands without the use of soap and water, and are thus an important tool in the practice of good hand hygiene. Alcohol-based hand rubs are recommended in healthcare settings for use on hands that are not visibly soiled. They are formulated to be applied full strength to dry hands, “rubbed in” until dry, and are not rinsed off.  
5.2 This test method is designed specifically to evaluate hand rubs for efficacy in eliminating bacteria from experimentally-contaminated hands. It is designed as an alternative to Test Method E1174, which was intended primarily to evaluate antimicrobial handwashing agents that are lathered with the aid of water and then rinsed off. When using Test Method E1174 to evaluate hand rubs, inadequate drying of the hands after contamination dilutes the test material and can compromise activity, to result in an underestimation of effectiveness. Additionally, because hand rubs are not rinsed after product use, activity can be further degraded by build-up of soil from the contaminating broth and inactivated challenge bacteria on the hands.  
5.2.1 In this method, application to the hands of a small volume of high-titer test bacteria suspension minimizes soil load such that the skin is completely dry prior to application of the test material. Further, by applying the bacterial suspension only prior to those test material application cycles followed by sampling, excessive buildup of killed bacteria on the hands is avoided, and the potential impact of non-volatile test product ingredients on bacteria-eliminating effectiveness after ten consecutive applications can be specifically assessed.  
5.3 A reference control is evaluated for each subject prior to evaluation of the test material. Data from the reference control helps to control for inter-subject variability, inter-experimental variabi...
SCOPE
1.1 This test method is designed to determine the activity of healthcare personnel hand rubs, (also known as hand rubs, hygienic hand rubs, hand sanitizers, or hand antiseptics) against transient microbial skin flora on the hands after a single application and after repeated applications.  
1.2 Performance of this procedure requires the knowledge of regulations pertaining to the protection of human subjects (see 21 CFR Parts 50 and 56).  
1.3 This test method should be performed by persons with training in microbiology, in facilities designed and equipped for work with potentially infectious agents at biosafety level 2.2,3  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For more specific precautionary statements, see 8.2.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method utilizes filter paper as a medium for evaluating touch-transfer of bacteria to material surfaces. Sample surfaces are exposed to filter papers saturated with a Staphylococcus aureus suspension, followed by recovery of the bacteria from the surface with Replicate Organism Detection and Counting (RODAC) plates. This test method reports the log10 reduction of bacterial transfer on an intended test surface compared to a control surface. The test and control surfaces can differ by texture, coating, treatment, or any other desired variables, as long as they are the same material.
SCOPE
1.1 This test method is designed to evaluate the contact-mediated microbial transference on two-dimensional material surfaces. Material surfaces intended to reduce microorganism adherence or contamination may be evaluated using this test method. Additionally, this test method can be utilized on a wide variety of material surfaces for other downstream processes.  
1.2 This test method is designed to quantify the fomite transfer of bacteria on material surfaces through touch-transfer, be inexpensive, require limited training in practice, and be adaptable to many surfaces or microorganisms. Non-porous surfaces are contaminated with inoculated filter paper as a carrier and surface contamination is quantified by sampling with Replicate Organism Detection and Counting (RODAC) plates.  
1.3 Basic microbiology training is required to perform this test method.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SCOPE
1.1 The purpose of this terminology standard is to establish uniformity in terms used in the field of agricultural chemical application. Terms are adopted from related fields and where applicable from Terminology E609.  
1.2 The terms are appropriate to any agricultural chemical application. Units in parenthesis following a definition are meant as typical and are not exhaustive of all units available for the term.  
1.3 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The Single Tube Method is designed to evaluate the efficacy of disinfectants against biofilm grown in the CDC biofilm reactor following the procedures outlined in Practice E3161. Biofilm grown in the CDC reactor is representative of biofilm that forms under high fluid shear on surfaces conducive to biofilm formation.  
5.1.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilm with specific characteristics (2). Altering either the engineered system or operating conditions will modify those characteristics as well as the physicochemical environment. The goal in biofilm research and testing is to choose the growth reactor and operating conditions that generate the most relevant biofilm for the particular study.  
5.2 The test method was designed to determine the log10 reduction in bacteria after exposure to a disinfectant in a closed system.  
5.3 The test method uses 50 mL conical tubes. The conical geometry allows for disinfectant exposure to biofilm on all surfaces of the coupon. For foaming disinfectants or for disinfectants requiring a larger volume of neutralizer, 250 mL conical tubes are used which preserve the required geometry and allow for greater neutralization capacity.  
5.4 Each test includes three untreated control coupons (exposed to buffered dilution water) and five treated coupons (per disinfectant/concentration/contact time combination).
SCOPE
1.1 This test method specifies the operational parameters required to perform a quantitative liquid disinfectant efficacy test against bacterial biofilm.  
1.2 The test method was optimized and validated for a Pseudomonas aeruginosa or Staphylococcus aureus biofilm grown in the CDC Biofilm Reactor (E3161). The method is suitable for evaluating additional bacteria grown using the procedures outlined in methods with comparable coupon dimensions such as Practice E3161, Test Method E2562, or Test Method E2196.  
1.3 Disinfectant preparation and contact time are used in the assessment according to the manufacturer’s instructions for use.  
1.4 The test method uses a closed system to treat biofilm. A coupon is placed in a single tube for the treatment, neutralization, and harvesting steps to prevent the loss of cells.  
1.5 This test method describes a harvesting and analysis procedure which includes vortexing and sonicating treated and untreated control biofilm, and recovery of culturable cells using filtration to lower the limit of detection. Biofilm population density is recorded as log10 colony-forming units per coupon. Efficacy is reported as a log10 reduction of culturable cells.  
1.6 Basic microbiology training is required to perform this assay.  
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Bacteria that exist in biofilms are phenotypically different from suspended cells of the same genotype. Research has shown that biofilm bacteria are more difficult to kill than suspended bacteria (4, 5). Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. The purpose of this practice is to direct a user in the growth of a P. aeruginosa or S. aureus biofilm by clearly defining the operational parameters to grow a biofilm that can be assessed for efficacy using the Standard Test Method for Evaluating Disinfectant Efficacy Against Pseudomonas aeruginosa Biofilm Grown in CDC Biofilm Reactor Using Single Tube Method (E2871).  
5.2 Operating the CDC Biofilm Reactor at the conditions specified in this method generates biofilm at log densities (log10 CFU per coupon) ranging from 8.0 to 9.5 for P. aeruginosa and 7.5 to 9.0 for S. aureus. These levels of biofilm are anticipated on surfaces conducive to biofilm formation such as the conditions outlined in this method.  
5.2.1 To achieve an S. aureus biofilm with a population comparable to that for P. aeruginosa using the bacterial liquid growth medium conditions specified here, the S. aureus biofilm must be grown at 36 °C ±2 °C rather than at room temperature (21 °C ±2 °C).
SCOPE
1.1 This practice specifies the parameters for growing a Pseudomonas aeruginosa (ATCC 15442) or Staphylococcus aureus (ATCC 6538) biofilm that can be used for disinfectant efficacy testing using the Test Method for Evaluating Disinfectant Efficacy Against Pseudomonas aeruginosa Biofilm Grown in CDC Biofilm Reactor Using Single Tube Method (E2871) or in an alternate method capable of accommodating the coupons used in the CDC Biofilm Reactor. The resulting biofilm is representative of generalized situations where biofilm exist on hard, non-porous surfaces under shear rather than being representative of one particular environment. Additional bacteria may be grown using the basic procedure outlined in this document, however, alternative preparation procedures for frozen stock cultures and biofilm generation (for example, medium concentrations, baffle speed, temperature, incubation times, coupon types, etc.) may be necessary.  
1.2 This practice uses the CDC Biofilm Reactor created by the Centers for Disease Control and Prevention (1).2 The CDC Biofilm Reactor is a continuously stirred tank reactor (CSTR) with high wall shear. The reactor is versatile and may also be used for growing or characterizing various species of biofilm, or both (2-4) provided appropriate adjustments are made to the growth media and operational parameters of the reactor.  
1.3 Basic microbiology training is required to perform this practice.  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this practice.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This test method is designed specifically for emulsions of pesticide emulsifiable concentrates.  
5.2 This test method provides information on the absorption/adsorption of solvents by simulated organic soil and inorganic soil impacted by pesticide EC emulsion applications.  
5.3 The amount of solvent lost by volatilization at 40 °C as determined by this method is an indirect measure of the atmospheric availability of the solvent to potentially react with nitrogen oxides to form tropospheric ozone, a major air pollutant.
SCOPE
1.1 This test method simulates the application of an emulsion of a pesticide emulsifiable concentrate (EC) to soil with high organic matter (corn cob granules) and to soil with high inorganic matter (clay granules) and determines the amount of solvent retained by the granules, and withheld from the atmosphere, before and after exposure to 40 ºC in a vented oven. The granules simulate two extremes of soil composition, and the 40 ºC exposure simulates high temperature weathering. Solvent loss from organic substrates other than corn cob may also be determined by repeating the 40 °C exposure tests with the chosen substrate replacing corn cob. The results with corn cob, however, are a reference that must be reported with the alternate substrate results. The difference in solvent content of the granules before and after weathering is an indication of the emission of the solvent from soil impacted by emulsions or solutions during pesticide applications using common practices such as spraying and drip irrigating. Analysis of the granules for solvent content is by high pressure liquid chromatography (HPLC), gas chromatography (GC), or other methods tested and proven to be accurate and reproducible.
Note 1: Since it evaluates soil surface sorption, this test method will underestimate soil sorption from pesticide applications made below the soil surface. Sub-soil surface treatments may include, but are not limited to, mechanical soil injection and soil incorporation applications. In these cases, the increased depth of the sub-soil treatments reduce the soil surface exposure and facilitate increased levels of soil sorption.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 The United States has concentrated attention and testing efforts on surgical scrubbing far more than on hand care in patient-to-patient routines. Great Britain, the originators of infection control nursing, have always had their focus on infection transmission. In the United States, published articles have documented the short exposure time for health care personnel who do wash their hands between patients. The average is less than 10 s. The ideal product for the reduction of transient flora is one that rapidly kills or removes or both the microbial load acquired during health care activities. The emphasis on rapidity is essential simply because health care personnel will not take the necessary time when using conventional hand-washing products. The use of products not intended for use with water has increased dramatically and their use is common in European countries largely because of convenience and effectiveness. A second characteristic is the level of antimicrobial action. The use of a rapid and potent active product to reduce work-acquired microbial flora is ideal.  
4.2 Since the change from strictly in-vitro testing of topical antimicrobials for use on skin to simulated use testing in hand washing, prepping, site access testing, and sampling, emphasis has always been on washing hands, agitating, rubbing, and brushing with liquid on the skin site to estimate bacteria removed after testing.  
4.3 The use of hard agitation has diminished with surgical scrubs without brushes or with only mild agitation and friction.  
4.4 There is a history of microbial dispersal (9) and increase in surface bacteria from deeper layers resulting from showering (10-12), washing, scrubbing, and agitation. In the normal situation on the skin, there is a superficial, surface flora and a deeper or hidden flora  (5). The proportion of one to the other has been addressed by Selwyn (4) and his judgment is that from 20 to 50 % of the flora is “deep.” The intent in skin sampling has...
SCOPE
1.1 This guide covers test methods and sampling procedure options for leave-on products for consumer and hospital personnel. Leave-on products, such as alcohol hand rubs and lotions containing antimicrobial ingredients, are increasingly marketed and used by consumers and health care personnel. These products are distinguished from conventional washing and scrubbing preparations in that they do not rely on the rinsing, physical removal, and antimicrobial action in determining their effectiveness. Although agitation and friction may serve to release organisms from the skin and folds and crevices, organisms are then killed in situ and are not rinsed from the skin surface before sampling. Appropriate test methods for the hands have been published, while other sampling methods will be needed for testing body areas other than the hands.  
1.1.1 Researchers have described techniques to identify the expanded flora we now know can be present on the skin. It is impractical, if not prohibitive to attempt to recover and identify these varieties of organisms with each test. At some point in the design of a test, a decision is necessary for defining the target organisms. Should the sampling be designed to recover as much of the microflora as possible or a particular portion of it? Consideration of transient and resident, superficial and deep, or aerobic and anaerobic flora must be included in defining the objective in testing products. The recovery methods selected for any testing must be based on the projected use of the product type being tested.  
1.2 Methods of recovery after application of the contaminating organisms to a part of the body other than by the agitation/rubbing of the hands against a glass petri plate also need examination. Consideration should be given to contact plating, controlled swabbing with a template, and cup scrubbing (detergent/agitation used) since the target organisms for recovery are likely to be on th...

SIGNIFICANCE AND USE
5.1 This practice is to help in the development of protocols to assess the survival, removal and/or inactivation of human pathogens or their surrogates in indoor air. It accommodates the testing of technologies based on physical (for example, UV light) and chemical agents (for example, vaporized hydrogen peroxide) or simple microbial removal by air filtration or a combination thereof.  
5.2 While this practice is designed primarily for work with aerobic, mesophilic vegetative bacteria, it can be readily adapted to handle other classes of microbial pathogens or their surrogates.  
5.3 The pieces of equipment given here are as examples only. Other similar devices may be used as appropriate.
SCOPE
1.1 This practice is to assess technologies for microbial decontamination of indoor air using a sealed, room-sized chamber (~24 m3) as recommended by the U.S. Environmental Protection Agency (3). The test microbe is aerosolized inside the chamber where a fan uniformly mixes the aerosols and keeps them airborne. Samples of the air are collected and assayed, firstly to determine the rates of physical and biological decay of the test microbe, and then to assess the air decontaminating activity of the technology under test as log10 or percentage reductions in viability per m3 (1). The air temperature and relative humidity (RH) in the chamber are measured and recorded during each test.  
1.2 The chamber can be used to assess microbial survival in indoor air as well as to test the ability of physical (for example, ultraviolet light) and chemical agents (for example, vaporized hydrogen peroxide) to inactivate representative pathogens or their surrogates in indoor air.  
1.3 This practice does not cover testing of microbial contamination introduced into the chamber as a dry powder.  
1.4 This practice does not cover work with human pathogenic viruses, which require additional safety and technical considerations.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 There are no reproducible standardized protocols for preparing specimens used to evaluate the microbicidal efficacy of non-chemical treatments such as ultraviolet (UV), highenergy electron beam, or other forms of non-chemical antimicrobial technologies.  
5.2 Conventional protocols for applying bioburdens to carriers (see Test Method E2197) cause cells to stack upon one another, thereby creating multiple cell layers in which cells in layers closer to the carrier are masked by cells in overlying layers, which makes relative comparison of different non-chemical antimicrobial treatments more difficult.  
5.3 Steel and other metal carriers have asperities that can shield a percentage of the applied cells from direct exposure to electromagnetic irradiation.  
5.4 The combined effects of 5.2 and 5.3 confound determination of the microbicidal effect of electromagnetic irradiation on test specimens.  
5.5 The practice addresses these two confounding factors by:  
5.5.1 Using glass microscope slides – the surfaces of which are asperity-free – as carriers.  
5.5.2 Reliably depositing bacterial cells onto the carrier as a monolayer.  
5.6 The resulting specimen ensures that all microbes deposited onto the carrier are exposed equally to the irradiation source thereby ensuring that the only variables are the controlled ones – starting inoculum concentration, wavelength (λ – in nm), exposure time(s), and resulting energy dose (J).
SCOPE
1.1 This practice provides a protocol for creating bacterial cell monolayers on a flat surface.  
1.2 The cultures used and culture preparation steps in this Practice are similar to AOAC Method 961.02 and US EPA MB-06. However, test bacteria are applied to the carrier using an automated deposition device (6.2) rather than as a suspension droplet.  
1.3 The carrier inspection protocol is similar to US EPA MB-03 except that carrier surfaces are inspected microscopically rather than visually, unaided.  
1.4 A monolayer of cells eliminates the confounding effect caused by the shadowing effect of outer layers of bacteria stacked upon other bacteria on test specimens – thereby attenuating directed energy beams (that is, ultraviolet light, high-energy electron beams) before they can reach underlying cells.  
1.5 An asperity-free surface eliminates the shadowing effect of specimen surface topology that can block direct exposure of target bacteria to non-chemical antimicrobial treatments.  
1.6 This practice provides a reproducible target microbe and surface specimen to minimize specimen variability within and between testing facilities. This facilitates direct data comparisons among various non-chemical antimicrobial technologies.  
1.6.1 Antimicrobial pesticides used in clinical and industrial applications are expected to overcome shadowing effects. However, this practice meets a need for a protocol that facilitates relative comparisons among non-chemical antimicrobial treatments.  
1.6.2 This practice is not intended to satisfy or replace existing test requirements for liquid chemical antimicrobial treatments (for example Test Methods E1153 and E2197) or established regulatory agency performance standards such as US EPA MB-06.  
1.7 This practice was validated using Staphylococcus aureus (ATCC 6538) and Pseudomonas aeruginosa (ATCC 15442) using a protocol based on AOAC Method 961.02. If other cultures are used, the suitability of this practice must be confirmed by inspecting prepared surfaces, by using scanning electron microscopy (SEM) or comparable high-resolution microscopy.  
1.8 The specimens prepared in accordance with this practice are not meant to simulate end-use conditions.  
1.8.1 Non-chemical technologies are only to be used on visibly clean, non-porous surfaces. Consequently, a soil load is not used.  
1.9 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.10 Th...

SIGNIFICANCE AND USE
5.1 In the battle to reduce medical device and implant-related infections, prevention of bacterial colonization of surfaces is a logical strategy. Bacterial colonization of a surface is a precursor to biofilm formation. Biofilm is the etiological agent of many implant and device-related infections and once established, microorganisms in biofilm can be up to 1000 times more tolerant to antibiotic therapy. Often the best treatment strategy is removal of the implant or device at a high socioeconomic cost. Catheter associated urinary tract infections (CAUTI) are the most prevalent of the device-related healthcare associated infections. Catheter associated infections account for 37 % of all hospital acquired infections (HAI) and 70 % of all nosocomial urinary tract infections (UTI) in the U.S. (2, 3). The Intraluminal Catheter Model (ICM) was developed to evaluate the ability of antimicrobial catheters to inhibit biofilm growth on the catheter lumen.  
5.2 The purpose of this test method is to direct a user in how to grow, sample, and analyze an E. coli biofilm in a urinary catheter under a constant flow of artificial urine. The test method incorporates operational parameters utilized in similar published methods (4). The E. coli biofilm that grows has a patchy appearance that varies across the catheter. Microscopically, the biofilm is heterogenous, with large clusters in some areas, and flat sheets of cells or even single cells in others. By 24 h, the biofilm is developed in the control catheters. If the goal is to monitor early stage biofilm development, then tubing and effluent samples need to be collected prior to the 24 h sample collection. Monitoring biofilm development requires sampling. The biofilm generated in the Intraluminal Catheter Model is suitable for comparison testing between antimicrobial and control catheters.
SCOPE
1.1 This test method specifies the operational parameters required to assess the ability of antimicrobial urinary catheters to prevent or control biofilm growth. Efficacy is reported as the log reduction in viable bacteria when compared to a repeatable (1)2  Escherichia coli biofilm grown in the intra-lumen of a urinary catheter under a constant flow of artificial urine.  
1.2 The test method is versatile and may also be used for growing and/or characterizing biofilms and suspended bacteria of different species, although this will require changing the operational parameters to optimize the method based upon the growth requirements of the new organism.  
1.3 This test method may be used to evaluate surface modified urinary catheters that contain no antimicrobial agent.  
1.4 This test method describes how to sample and analyze catheter segments and effluent for viable cells. Biofilm population density is recorded as log colony forming units per surface area. Suspended bacterial population density is reported as log colony forming units per volume.  
1.5 Basic microbiology training is required to perform this test method.  
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard  
1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This procedure was designed principally for clay, corncob, nut shell, paper, or sand granular carriers and granular pesticide products, but need not be limited to these materials. There may be more appropriate test methods for other types of granular carriers and products.
SCOPE
1.1 This test method is used to determine the particle size distribution of granular carriers and granular pesticides.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific hazard statements, see Section 6.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The procedures in this practice should be used for in vivo evaluation of the antimicrobial activity of drug products applied topically to the skin that are intended to help prevent infection in minor cuts, scrapes and burns.  
5.1.1 This practice is applicable for testing liquids, ointments, powders, films, or dressings, containing or impregnated with an antimicrobial agent, for their effect to reduce an enhanced skin microflora or their effects to suppress the growth of the skin flora, or both.
SCOPE
1.1 The tests described in this practice are designed to evaluate antimicrobial agents in formulations intended for use as first aid antiseptic products for their ability to reduce or suppress the growth, or both, of the skin microflora.  
1.2 A knowledge of microbiological techniques is required for these procedures.  
1.3 Performance of this procedure requires the knowledge of regulations pertaining to the protection of human subjects. (See CFR Parts 50 and 56.)  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method is designed specifically for dry formulations, but need not be restricted to these materials.  
4.2 Either option of this test method (see Section 8) may be used to determine the percent suspensibility.  
4.3 This test method may not be applicable to all dry formulations such as those containing liquid or soluble technical/pesticides or ingredients that rise to the top upon separation.  
4.4 This test method may not be applicable to formulations containing ingredients that decompose under the test conditions.  
4.5 This test method may not give reproducible results if flocculation occurs.  
4.6 This test method should be run in duplicate.  
4.7 Products containing water soluble or volatile components may result in errors.
SCOPE
1.1 This test method is used to determine the percent suspensibility of dry pesticide formulations.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific precautionary statements see Section 7.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method is designed specifically for dry formulations.  
4.2 This test method may not be applicable to all dry formulations such as those containing water soluble or liquid technicals/pesticides or ingredients that rise to the top upon separation.  
4.3 This test method may not be applicable to those technicals that decompose below the drying temperature.  
4.4 This test method should be run in duplicate.  
4.5 Products containing water soluble or volatile components may result in errors.
SCOPE
1.1 This test method is used to determine the percent dispersibility of dry pesticide formulations.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  For specific precautionary statements see Section 7.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This test method is intended for use in quality control, material screening, and related problem solving where a nonvolatile content is desired or a comparison can be made with a known material of the same type.  
5.2 The parameters described may be altered to suit a particular analysis, provided the changes are noted in the report.
SCOPE
1.1 This test method covers the determination of the non-volatile matter of agricultural tank mix adjuvant solutions by thermogravimetry.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The different procedures and methods are designed to be used to produce survival data after microorganisms are exposed to antimicrobial agents in order to calculate values that can be used to analyze and rationalize the effectiveness of antimicrobial agents when tested using other, often applied test methods.  
5.2 The data from these test procedures may be used in the selection and design of other tests of effectiveness of antimicrobial agents, some of which may be required by regulatory agencies to establish specific claims. Basic kinetic information about killing rate often serves as the initial information on which a testing program can be built.
SCOPE
1.1 This guide covers the methods for determining the death rate kinetics expressed as D-values. These values can be derived from the construction of a kill curve (or survivor curve) or by using other procedures for determining the number of survivors after exposure to antimicrobial chemicals or formulations. Options for calculations will be presented as well as the method for calculation of a concentration coefficient.  
1.1.1 The test methods are designed to evaluate antimicrobial agents in formulations to define a survivor curve and to subsequently calculate a D-value. The tests are designed to produce data and calculate values that provide basic information of the rate-of-kill of antimicrobial formulations tested against single, selected microorganisms. In addition, calculated D-values from survivor curves from exposure at different dilutions of antimicrobial can be used to show the effect of dilution by calculation of the concentration exponent, η (2). D-value determination assumes the ideal of first-order killing reactions that are reflected in a straight-line reduction in count where a count-versus-time plot is done. The goal here is not to determine the time at which no survivors are found, but to determine a standard value that can be used in processing and exposure determinations or used to estimate dilutions.  
1.1.2 As an example of potential use of kill curve data, the published FDA, OTC Tentative Final Monograph for Health-Care Antiseptic Drug Products, Proposed Rule, June 17, 1994 has suggested the testing of topically applied antimicrobial products using survival curve (or kill curve) calculations. The methods described in this guide are applicable to these products, but adjustments such as the use of antifoaming agents when the reaction mixture is stirred may be necessary to counteract the presence of detergents in many formulations. Frequently the sampling for these tests is done after very short intervals of exposure to the formulation, such as 30 and 60 s. This methodology also has been applied to preservative testing of antimicrobial ingredients in more complex cosmetic formulations (5).  
1.2 The test methods discussed should be performed only by those trained in microbiological techniques.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The procedure is used to test the antimicrobial effectiveness of handwashing formulations. The test formulations generally are designed for frequent use to reduce the transient bacterial flora on hands. Alcohol-based hand rubs and other leave-on formulations used without the aid of water should be tested using Test Method E2755.
SCOPE
1.1 This test method is designed to determine the effectiveness of antimicrobial handwashing agents for the reduction of transient microbial skin flora when used in a handwashing procedure.2  
1.2 A knowledge of microbiological techniques is required for these procedures.  
1.3 This test method may be used to evaluate topical antimicrobial handwash formulations.  
1.4 Performance of this procedure requires the knowledge of regulations pertaining to the protection of human subjects.3  
1.5 The values stated in SI units are to be regarded as standard; except for distance, in which case inches are used and metric units follow in parentheses.  
1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.For more specific precautionary statements see 8.2.  
1.7 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This practice describes a procedure for preparing and storing a suspension of C. difficile spores that meets the following acceptance criteria: (1) spore titer of approximately 5.0×108 spores/mL, (2) spore purity of ≥95 %, and (3) a mean log10 reduction (LR) value >5.0 for 3 carriers exposed to 5000 ppm and a mean LR of E3218.
SCOPE
1.1 This practice specifies the procedures for producing and storing standardized suspensions of Clostridioides difficile spores for the evaluation of the sporicidal activity of antimicrobial formulations using the Quantitative Method for Testing Antimicrobial Agents against Spores of C. difficile on Hard, Non-porous Surfaces or other procedures.  
1.2 This practice may involve hazardous materials, chemicals, and microorganisms and should be performed only by persons with formal training in microbiology.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 The test method was designed to determine the LR in spores on a hard, non-porous surface after exposure to a test chemical in a closed system.  
5.2 Each test includes three control carriers (exposed to phosphate buffered saline with Tween-80), three test system control carriers (exposed to 1500 ppm ± 150 ppm sodium hypochlorite), and ten treated carriers (per test chemical/concentration/contact time combination).
SCOPE
1.1 This test method covers a standardized approach to quantitatively determine the effectiveness of antimicrobial chemicals in treating hard, non-porous surfaces contaminated with spores of C. difficile (ATCC 43598) grown in accordance with Practice E2839.  
1.2 This test method is based on principles established for Test Method E2197 and an Organisation for Economic Co-operation and Development Guidance Document.2  
1.3 Training in basic microbiology and aseptic technique are required to perform this assay.  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
4.1 This test method is intended for products sold as tank mix adjuvants for use with agricultural products.  
4.2 Part A provides a determination of whether the pH modification characteristics of the adjuvant tested meet the definition of an acidifier, alkalinity agent, or a basic blend in Terminology E1519. Part B provides a measure of the buffer capacity of the adjuvant. This can be used to estimate the amount of a buffer that will be required to keep a spray solution at the desired pH.  
4.3 Other pH standards can be used (see Test Method E70) as long as the pHs measured are not outside the range of the standards used.  
4.4 If samples of the material to be used in a particular application are available, the first method can be used to determine what adjuvant rate is required to obtain the desired pH. The ability of an adjuvant to adjust the pH of a particular system will depend on the initial pH before addition.  
4.5 The second method can be used to determine the amount of an adjuvant required to change the pH of a quantity of spray solution to the required value (see Note 1).
Note 1: Many acidifiers and basic blends also are buffers, so the pH change does not provide a useful measure of buffer capacity.
SCOPE
1.1 These test methods cover determining if an adjuvant meets the definitions of an acidifier, a basic blend, alkalinity agent, or buffer in Terminology E1519.  
1.2 The values stated in either SI units or inch-pound units are to be regarded separately as standard. The values stated in each system may not be exact equivalents; therefore, each system shall be used independently of the other. Combining values from the two systems may result in non-conformance with the standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 Hand hygiene is considered one of the most important measures for preventing the spread of infectious microorganisms and is critical for reducing the incidence of food-borne disease. Food-handling settings are unique in that moderate to heavy soil load present on hands often can influence the ability of a product to remove or kill microorganisms (3, 4). Test methods are needed for assessing the efficacy of hand hygiene products under conditions representative of those encountered in a food-handling environment.  
5.2 This test method is specifically designed to evaluate the effectiveness of food-handler products to kill and remove bacteria from experimentally-contaminated hands under conditions of moderate to heavy organic soil load. The inclusion of soils typical of food service setting makes this a methodology more appropriate than Test Method E1174, which was designed to evaluate healthcare personnel hand washes and does not include an option to include soil (4).
SCOPE
1.1 This test method is designed to determine the activity of food-handler handwashes against transient bacterial flora on the hands.  
1.2 Performance of this procedure requires the knowledge of regulations pertaining to the protection of human subjects (1)2.  
1.3 This test method should be performed by persons with training in microbiology, in facilities designed and equipped for work with potentially infectious agents at biosafety level 2 (2).  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For more specific precautionary statements see 8.1.1.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
3.1 This procedure was designed principally for clay, corncob, nut shell, or sand granular carriers and granular pesticide products but need not be limited to these materials. There may be more appropriate sampling methods for other types of granular carriers and products.
SCOPE
1.1 This test method presents general procedures for sampling and sampling practice for granular carriers and granular pesticides.  
1.2 The values stated in inch-pound units are to be regarded as standard. The values given in parentheses are mathematical conversions to SI units that are provided for information only and are not considered standard.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific hazard statements see Section 5.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

SIGNIFICANCE AND USE
5.1 This test method provides a simple approach for determining the transmission properties of film membranes and sheeting over a range of permeability exceeding four orders of magnitude. This test method is described here to measure the permeability of films used in soil fumigation, but it is also appropriate for other gases and membranes if the analytical methods are appropriately modified.  
5.2 This test method can be used for single or mixed compounds. This test method uses small quantities of test chemicals in vapor form, and microgram to milligram quantities of each chemical may produce a sufficient amount of vapor for each test depending on the analytical methods.  
5.3 Interlaboratory testing showed that the MTC estimated by this test method is relatively insensitive to the laboratory procedures. The interlaboratory testing involved measuring the MTC for several soil fumigant compounds and a wide range of film permeability. Analysts with prior experience handling and analyzing gaseous fumigant compounds had lower coefficients of variation (10 % to 20 %) compared to less experienced analysts (20 % to 50 %) based on triplicate tests. The coefficient of variation between laboratories was higher for less permeable film materials than for films with high MTC. This was attributed to the additional length of the experiments and potential for increased leakage from the apparatus and was most pronounced for less experienced analysts.
SCOPE
1.1 This test method covers the measurement of the transmission of a gas through plastic membranes, sheeting, films, and fabric materials using a static sealed diffusion chamber. The test method monitors gas diffusion across a film membrane and provides measurements of (1) gas concentrations on each side of the film membrane and (2) estimates of the mass transfer coefficient (MTC) for the tested gas and film material. The MTC represents the film permeability and is independent of the concentration gradient used during testing, which simplifies some aspects of the experimental design.  
1.2 This test method permits the loading of mixed vapors and simultaneous determination of the permeability of one film to various gases.  
1.3 Units—The values stated in SI units are to be regarded as the standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.