ASTM E2199-02
(Specification)Standard Specification for Encarsia formosa Gahan (Hymenoptera:Aphelinidae)
Standard Specification for <i>Encarsia formosa</i> Gahan (<i>Hymenoptera:Aphelinidae</i>)
SCOPE
1.1 This specification covers information on and the test method for determining the purity and number of adults released in shipments of Encarsia formosa, parasite of the greenhouse whitefly, Trialeurodes vaporariorum Westwood.
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E 2199 – 02
Standard Specification for
Encarsia formosa Gahan (Hymenoptera:Aphelinidae)
This standard is issued under the fixed designation E 2199; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope 5.2 The number of E. formosa adults released from parasit-
ized whitefly scales attached to cards will be determined by
1.1 This specification covers information on and the test
examining a minimum of 1000 parasitized scales and estimat-
method for determining the purity and number of adults
ing total number of live E. formosa and 95 % Confidence
released in shipments of Encarsia formosa, parasite of the
Limits (C.L.) per shipment. In each container, live contami-
greenhouse whitefly, Trialeurodes vaporariorum Westwood.
nants will be identified and recorded.
2. Referenced Documents
6. Significance and Use
2.1 ASTM Standards:
6.1 The biological control of greenhouse whitefly by E.
E 2200 Specification for Information Included With Pack-
formosa depends on accurate release numbers of adult para-
aging of Multi-cellular Biological Control Organisms
sites. Accurate packaging and maintenance of purity and
3. Terminology
viability of E. formosa shipments is, therefore, essential for the
effective management of this pest. This test method may be
3.1 Definitions of Terms Specific to This Standard:
performed by the producers and end-users.
3.1.1 life stage when shipped—immature.
3.1.2 name of product—Encarsia formosa Gahan
7. Materials
3.1.3 preferred host prey—greenhouse whitefly, Trialeu-
7.1 Dissecting microscope or headband magnifier (7 to
rodes vaporariorum (Westwood)
103).
4. Classification
7.2 45 mm diameter by 90 mm deep, clear plastic vial with
snap top.
4.1 Phylum—Arthropoda.
7.3 Fine pointed No. 5/0 paint brush.
4.2 Class—Insecta.
4.3 Order—Hymenoptera.
8. Test Unit
4.4 Family—Aphelinidae.
8.1 A single shipment of E. formosa is considered a test
4.5 Genus—Encarsia.
unit.
4.6 Species—formosa.
9. Pre-Test Conditions
TEST METHOD—Determining the Purity and Number
of Adults Released in Shipments of E. formosa on Cards 9.1 If samples must be held before testing, hold them
between 10 and 15°C, RH 60 to 90 %, for a maximum of 24 h.
5. Summary of Test Method
10. Sampling
5.1 This test method describes a method of counting the
number of E. formosa released from cards. 10.1 Specify the number of adults that should emerge from
individual cards as indicated on the package before conducting
the test. Randomly select enough cards per shipment to make
up a minimum of 1000 parasites per test unit.
This specification is under the jurisdiction of ASTM Committee E35 on
Pesticides and Alternative Control Agents and are the direct responsibility of
11. Sample Preparation and Treatment
Subcommittee E35.30 on Natural Multi-Cellular (Metazoan) Biological Control
Organisms.
11.1 Close each individual E. formosa card in a 45 mm
Current edition approved April 10, 2002. Published July 2002.
diameter by 90 mm deep, clear plastic vial. Cut a circle of
Steinberg, S., and Dale, J., “Designing and Implementing Quality Control of
yellow double-sided sticky trap card the same size as the inside
Beneficials Insects: Towards More Reliable Biological Pest Control,” Sting,
Newsletter on Biologi
...
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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.
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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)).
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.
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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.
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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.
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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.
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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.
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SIGNIFICANCE AND USE
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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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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.
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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.
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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.
1.2 This test method allows for the ability to evaluate many different types of treated substrates and a wide range of microorganisms. Treated substrates used in this test method can be subjected to a wide variety of physical/chemical stresses or manipulations and allows for the versatility of testing the effect of contamination due to such things as hard water, proteins, blood, serum, various chemicals, and other contaminants.
1.3 Surface antimicrobial activity is determined by comparing results from the test sample to controls run simultaneously.
1.4 This test method may not be appropriate for all types of antimicrobial-treated articles or antimicrobial agents. The proper test methodology should be determined based on antimicrobial mode of action and end-use expectations (Guide E2922)
1.5 Proper neutralization of all antimicrobials must be confirmed using Test Methods E1054.
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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SIGNIFICANCE AND USE
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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.
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