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ASTM E2196-07

(Test Method)

Standard Test Method for Quantification of a Pseudomonas aeruginosa Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk Reactor

Standard Test Method for Quantification of a <i>Pseudomonas aeruginosa</i> Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk Reactor

SIGNIFICANCE AND USE
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.
SCOPE
1.1 This test method is used for growing a repeatable Pseudomonas aeruginosa biofilm in a continuously stirred flow reactor. In addition, the test method describes how to sample and analyze biofilm for viable cells.
1.2 In this test method, biofilm population density is recorded as log colony forming units per surface area.
1.3 Basic microbiology training is required to perform this test method. This standard does not claim to address all of the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety practices and determine the applicability of regulatory limitations prior to use.

General Information

Status
Historical
Publication Date
31-Mar-2007
ICS
07.100.01 - Microbiology in general
Technical Committee
E35 - Pesticides, Antimicrobials, and Alternative Control Agents
Drafting Committee
E35.15 - Antimicrobial Agents
Current Stage
Ref Project

Relations

Replaces
ASTM E2196-02 - Standard Test Method for Quantification of a <i>Pseudomonas aeruginosa</i> Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk Reactor
Effective Date
01-Apr-2007
Replaced By
ASTM E2196-12 - Standard Test Method for Quantification of <i>Pseudomonas aeruginosa</i> Biofilm Grown with Medium Shear and Continuous Flow Using Rotating Disk Reactor
Effective Date
01-Apr-2012
Referred By
ASTM E2871-21 - Standard Test Method for Determining Disinfectant Efficacy Against Biofilm Grown in the CDC Biofilm Reactor Using the Single Tube Method
Effective Date
01-Apr-2007

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ASTM E2196-07 - Standard Test Method for Quantification of a <i>Pseudomonas aeruginosa</i> Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk ReactorASTM E2196-07 - Standard Test Method for Quantification of a <i>Pseudomonas aeruginosa</i> Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk Reactor
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ASTM E2196-07 - Standard Test Method for Quantification of a <i>Pseudomonas aeruginosa</i> Biofilm Grown with Shear and Continuous Flow Using a Rotating Disk Reactor
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Please contact ASTM International (www.astm.org) for the latest information.
Designation: E2196 – 07
Standard Test Method for
Quantification of a Pseudomonas aeruginosa Biofilm Grown
with Shear and Continuous Flow Using a Rotating Disk
1
Reactor
This standard is issued under the fixed designation E2196; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope other, embedded in a matrix of extracellular polymeric sub-
2 stances of microbial origin, while exhibiting an altered pheno-
1.1 This test method is used for growing a repeatable
type with respect to growth rate and gene transcription.
Pseudomonasaeruginosabiofilminacontinuouslystirredflow
3.1.1 Discussion—Biofilms may be comprised of bacteria,
reactor. In addition, the test method describes how to sample
fungi, algae, protozoa, viruses, or infinite combinations of
and analyze biofilm for viable cells.
these microorganisms. The qualitative characteristics of a
1.2 In this test method, biofilm population density is re-
biofilm, including, but not limited to, population density,
corded as log colony forming units per surface area.
taxonomic diversity, thickness, chemical gradients, chemical
1.3 Basic microbiology training is required to perform this
composition,consistency,andothermaterialsinthematrixthat
test method. This standard does not claim to address all of the
arenotproducedbythebiofilmmicroorganisms;arecontrolled
safety problems associated with its use. It is the responsibility
by the physiochemical environment in which it exists.
of the user of this standard to establish appropriate safety
3.2 coupon—biofilm sample surface.
practices and determine the applicability of regulatory limita-
tions prior to use.
4. Summary of Test Method
2. Referenced Documents 4.1 This test method is used for growing a repeatable
Pseudomonas aeruginosa biofilm in a rotating disk reactor.
2.1 Other Standards:
3
The biofilm is established by operating the reactor in batch
Buffered Dilution Water Preparation —Method 9050 C.1a
4
mode (no flow) for 24 h. A steady state growth (attachment is
Rotating Disk Reactor —Repeatability and Relevance
5
equal to detachment) is reached while the reactor operates for
Rotating Disk Reactor —Efficacy Test Method
an additional 24 h with continuous flow of the nutrients. The
3. Terminology
residence time of the nutrients in the reactor is set to select for
biofilm growth, and is species and reactor parameter specific.
3.1 biofilm, n—microorganisms living in a self-organized,
During the entire 48 h, the biofilm is exposed to continuous
cooperativecommunityattachedtosurfaces,interfaces,oreach
fluid shear from the rotation of the disk.At the end of the 48 h,
biofilm accumulation is quantified by removing coupons from
1
This test method is under the jurisdiction of ASTM Committee E35 on
the disk, scraping the biofilm from the coupon surface,
Pesticides and Alternative Control Agents and is the direct responsibility of
disaggregating the clumps, then diluting and plating for viable
Subcommittee E35.15 on Antimicrobial Agents.
cell enumeration.
Current edition approved April 1, 2007. Published May 2007. Originally
approved in 2002. Last previous edition approved in 2002 as E2196 – 02. DOI:
10.1520/E2196-07.
5. Significance and Use
2
Ellison, S.L.R., M. Rosslein, A. Williams. (Eds.) 2000. Quantifying Uncer-
5.1 Bacteria that exist in a biofilm are phenotypically
tainty in Anyalytical Measurement, 2nd Edition. Eurachem.
3
Eaton, A.D., L.S. Clesceri, Rice, E.W., A.E. Greenberg. (Eds.) Standard different from suspended cells of the same genotype.The study
Methods for the Examination of Water and Waste Water , 21st Edition. American
of biofilm in the laboratory requires protocols that account for
Public HealthAssociation,American Water WorksAssociation, Water Environment
this difference. Laboratory biofilms are engineered in growth
Federation. Washington D.C., 2005.
4
reactors designed to produce a specific biofilm type. Altering
Zelver, N., M. Hamilton, B. Pitts, D. Goeres, D. Walker, P. Sturman, J.
Heersink. 1999. Methods for measuring antimicrobial effects on biofilm bacteria:
system parameters will correspondingly result in a change in
from laboratory to field. In: Doyle, R.J. (Ed.), Methods in Enzymology-Biofilms Vol
thebiofilm.Thepurposeofthismethodistodirectauserinthe
310, Academic Press, San Diego, CA, pp. 608-628.
5 laboratory study of biofilms by clearly defining each system
Zelver, N., M. Hamilton, D. Goeres, J. Heersink. 2001. Development of a
Standardized Antibiofilm Test. In: Doyle, R.J. (Ed.), Methods in Enzymology-
Biofilms Vol 337, Academic Press, San Diego, CA, pp. 363-376.
Copyright © ASTM International, 100 Barr Harbor Dri
...

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Standard Test Method for Quantification of Pseudomonas aeruginosa Biofilm Grown with Medium Shear and Continuous Flow Using Rotating Disk Reactor

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.
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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)).  
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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.  
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Standard Test Method for Testing Disinfectant Efficacy against Pseudomonas aeruginosa Biofilm using the MBEC Assay

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  
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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.  
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Standard Practice for Preparing a Pseudomonas aeruginosa or Staphylococcus aureus Biofilm using the CDC Biofilm Reactor

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).
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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.  
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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.  
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5.3 The pieces of equipment given here are as examples only. Other similar devices may be used as appropriate.
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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.  
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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.  
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1.5 Basic microbiology training is required to perform this test method.  
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Standard Practice for Preparation Of Cell Monolayers on Glass Surfaces for Evaluation of Microbicidal Properties of Non-Chemical Based Antimicrobial Treatment Technologies

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.  
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SIGNIFICANCE AND USE
5.1 Substrate–bonded, antimicrobial agents are not typically free to diffuse into their environment under normal conditions of use. This test method ensures good contact between the bacteria and the treated fiber, fabric, or other substrate, by constant agitation of the test specimen in a challenge suspension during the test period.  
5.2 The metabolic state of the challenge species can directly affect measurements of the effectiveness of particular antimicrobial agents or concentrations of agents. The susceptibility of the species to particular biocides could be altered depending on its life stage (cycle). One-hour contact time in a buffer solution allows for metabolic stasis in the population. This test method standardizes both the growth conditions of the challenge species and substrate contact times to reduce the variability associated with growth phase of the microorganism.  
5.3 Leaching of an antimicrobial is dependent upon the test conditions being utilized and the ultimate end use of the product. Additional testing may be required to determine if a compound is substrate-bound in all conditions or during the end use of the product.  
5.4 This test method cannot determine if a compound is leaching into solution or is immobilized on the substrate. This test method is only intended to determine efficacy as described in subsequent portions of the method.  
5.5 The test is suitable for evaluating stressed or modified specimens, when accompanied by adequate controls.
Note 1: Stresses may include laundry, wear and abrasion, radiation and steam sterilization, UV exposure, solvent manipulation, temperature susceptibility, or similar physical or chemical manipulation.
SCOPE
1.1 This test method is designed to evaluate the antimicrobial activity of antimicrobial-treated specimens under dynamic contact conditions. This dynamic shake flask test was developed for routine quality control and screening tests in order to overcome difficulties in using classical antimicrobial test methods to evaluate substrate-bound antimicrobials. These difficulties include ensuring contact of inoculum to treated surface (as in AATCC TM100), flexibility of retrieval at different contact times, use of inappropriately applied static conditions (as in AATCC TM147), sensitivity, and reproducibility.  
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1.6 This test method should be performed only by those trained in microbiological techniques.  
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 may involve hazardous materials, operations and equipment. 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.  
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    English language
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  • Standard
    5 pages
    English language
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ASTM
ASTM E3226-19(Test Method)
Standard Test Method for Processing Cellulose Sponge-wipes to Detect Bacillus anthracis Spores Sampled from Environmental Surfaces

SIGNIFICANCE AND USE
5.1 This procedure describes a standardized method of processing cellulose wipes in a biosafety level 3 laboratory in order to detect and provide a semi-quantitative estimate of B. anthracis contamination after sampling of non-porous surfaces. Sampling may be conducted to characterize the extent of contamination or for clearance of an area after decontamination.
SCOPE
1.1 This test method covers a standardized method of processing cellulose wipes in a biosafety level 3 (BSL3) laboratory in order to detect and provide a semi-quantitative estimate of Bacillus anthracis contamination after sampling of non-porous surfaces. Sampling may be conducted to characterize the extent of the contamination, or for area clearance after decontamination.  
1.2 The laboratory procedures should be performed in a BSL3 laboratory by those trained for BSL3 microbiological techniques.  
1.3 This test method is specific to B. anthracis, but could be adapted for use with other organisms.  
1.4 The interlaboratory study was conducted with cellulose sponge wipes pre-moistened with neutralizing buffer. All reproducibility, sensitivity, and specificity data are based on the performance of these wipes. A review was conducted by subcommittee in 2019, and re-confirmed these ILS data are valid.  
1.5 Units—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.  
1.6 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.

  • Standard
    14 pages
    English language
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ASTM
ASTM E3179-18(Test Method)
Standard Test Method for Determining Antimicrobial Efficacy of Ultraviolet Germicidal Irradiation against Influenza Virus on Fabric Carriers with Simulated Soil

SIGNIFICANCE AND USE
5.1 This test method determines the effectiveness of UVGI devices for reducing viable microorganisms deposited on carriers.  
5.2 This test method evaluates the effect soiling agents have on UVGI antimicrobial effectiveness.  
5.3 This test method determines the delivered UVGI dose.
SCOPE
1.1 This test method defines test conditions to evaluate ultraviolet germicidal irradiation (UVGI) light devices (mercury vapor bulbs, light-emitting diodes, or xenon arc lamps) that are designed to kill/inactivate influenza virus deposited on inanimate carriers.  
1.2 This test method defines the terminology and methodology associated with the ultraviolet (UV) spectrum and evaluating UVGI dose.  
1.3 This test method defines the testing considerations that can reduce UVGI surface kill effectiveness (that is, soiling).  
1.4 Protocols for adjusting the UVGI dose to impact the reductions in levels of viable influenza virus are provided (Annex A1).  
1.5 This test method does not address shadowing.  
1.6 The test method should only be used by those trained in microbiology and in accordance with the guidance provided by Biosafety in Microbiological and Biomedical Laboratories.2  
1.7 This test method is specific to influenza viruses  
1.8 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.9 Warning—Mercury has been designated by many regulatory agencies as a hazardous substance that can cause serious medical issues. Mercury, or its vapor, has been demonstrated to be hazardous to health and corrosive to materials. Use caution when handling mercury and mercury-containing products. See the applicable product Safety Data Sheet (SDS) for additional information. The potential exists that selling mercury or mercury-containing products, or both, is prohibited by local or national law. Users must determine legality of sales in their location.  
1.10 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.  
1.11 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.

  • Standard
    6 pages
    English language
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