Designation: E2753 − 10Standard Specification forEretmocerus eremicus Rose and Zolnerowich(Hymenoptera: Aphelinidae)1This standard is issued under the fixed designation E2753; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (´) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This specification describes a method for determiningwhether the quantity and quality of adult Eretmocerus eremicusin a shipment adhere to quantity and quality specifications. Thetest also allows the purity of shipments to be determined.Included are referenced documents, a description of standardterminology, specifications, and the test method.1.2 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.2. Referenced Documents2.1 ASTM Standards:2E2200 Specification for Information Included with Packag-ing of Multi-Cellular Biological Control Organisms(Withdrawn 2010)33. Terminology3.1 name of product—Eretmocerus eremicus Gahan.3.2 preferred hosts and prey—Greenhouse Whitefly, Tri-aleurodes vaporariorum (Westwood), Silverleaf Whitefly, Be-misia argentifolii Bellows and Perring and Tobacco Whitefly,Bemisia tabaci (Gennadius).3.3 life stage shipped—pupa within the host insect, T.vaporariorum.3.4 card—a sample unit. Pupae of Eretmocerus eremicus aremounted on cardstock strips that are subdivided into “cards”.3.5 package claim—the number of adults expected toemerge from and leave each card.3.6 test statistic—the average number of wasps caughtduring the flight test expressed as a percentage of the packageclaim.3.7 critical value—a number that the test statistic is com-pared to in order to determine whether the quantity and qualityrequirement has been met. Critical values depend on theprobability of error that can be tolerated and on the number ofsamples used in the test.4. Classification4.1 Phylum—Arthropoda.4.2 Class—Insecta.4.3 Order—Hymenoptera.4.4 Family—Aphelinidae.4.5 Genus—Eretmocerus.4.6 Species—eremicus.5. Summary of Test Method (Determining the Number ofE. eremicus Released from a Shipment and itsConformity to the Standard)5.1 The test describes methods for determining whether thenumber of adult E. eremicus that are capable of flight meets orexceeds the package claim.Amethod of assessing the purity ofa shipment is included.5.2 The quantity of E. eremicus per card and their flightability is determined by counting the number of wasps caughton a sticky trap suspended in a test chamber. Three or more testchambers are used per shipment, each chamber contains onerandomly chosen card with E. eremicus pupae. The numbers ofwasps caught on each sticky card are counted and results areused to calculate the average number of wasps per card. Thisaverage is used in combination with the known samplingdistribution and variation in counts to judge whether specifi-cations are met. Live contaminants are identified and recorded.5.3 Quantity and Quality Specification—The test describedin this document is used for both the quantity and qualityassessment. To meet the quantity and quality specificationdefined by the “package claim”, 100 % of the number of adultsshould be capable of flight as measured by the flight testdescribed herein.1This specification is under the jurisdiction of ASTM Committee E35 onPesticides, Antimicrobials, and Alternative Control Agents and is the directresponsibility of Subcommittee E35.30 on Natural Multi-Cellular (Metazoan)Biological Control Organisms.Current edition approved April 1, 2010. Published May 2010. DOI: 10.1520/E2753–10.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at

[email protected] For Annual Book of ASTMStandards volume information, refer to the standard’s Document Summary page onthe ASTM website.3The last approved version of this historical standard is referenced onwww.astm.org.Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States15.4 Purity—The purity specification is that shipments, andhence samples, do not contain species other than Eretmoceruseremicus.6. Significance and Use6.1 This method was developed to determine that thenumbers of E. eremicus supplied in a shipment meet thepackage claim and that wasps at receipt have good flightcapability. The application of this method will ensure astandardized evaluation of the product and judicious decisionsabout product compliance to the package claim.7. Materials7.1 Test chamber (6.5×8cmacrylic tube).7.2 Support wire.7.3 Double-sided dry-stick yellow sticky trap (2.5 × 4 cm).7.4 Nylon screen (100 micron mesh).7.5 Clear plastic cling-film (see Fig. 1).7.6 Headband magnifier (7 to 10×) is optional.8. Test Unit8.1 A single shipment of E. eremicus is considered a testunit.9. Pre-Test Conditions9.1 If required, a shipment can be stored for a maximum 24h at 10 to 15°C, relative humidity (RH) 60 to 90 %.10. Sample Size10.1 Choose three or more sample cards per shipment (seethe interpretation of results to decide on the sample size).11. Sampling and Testing11.1 Place one card on the floor of each chamber. Positionthe yellow sticky trap inside the chamber on the support wire.Seal the top of the chamber with plastic wrap. Place thechamber within the crop, away from the heating pipes and outof direct sunlight. Test conditions should be within 20 to 25°Cand 60 to 90 % RH.12. Counting Procedure and the Assessment of Purity12.1 After two weeks, count the number of wasps caught onthe sticky trap and calculate the average for all chambers used.Express the average as a percentage of the number expected(the package claim) from the card. This value is called the teststatistic.12.2 Examine the parasitized scales and the yellow stickytrap and record the presence of other live insects or mitesappearing among the parasitoids.13. Properties of the Test13.1 In the absence of other data, the quantity and quality ofwasps in shipments are assumed to meet or exceed packageclaims. The test statistic and known properties of the counts ofwasps on sticky cards are used to judge whether the packageclaim is met. This is done by comparing the test statistic to acritical value (CV) that is tied to a probability of incorrectlydeclaring a shipment to be substandard and to the number ofcards tested. The CVs for three sample sizes and differentprobabilities of error are presented in Table 2.ACVisdetermined by selecting a sample size (the number of cardsassessed) and a probability of error. For example, if 3 cardswere assessed and a probability of error of 0.05 was chosen,CV = 90.13.2 Each probability of error in Table 2 is a measure of thelikelihood of wrongly categorizing the package to be deficientwhen it in fact has the specified number of healthy, flightcapable insects. More precisely, these are the probabilities ofobtaining a test statistic that is less than or equal to the CVwhen the package claim is exactly met. Note that for aparticular CV, the probability of error declines with increasingsample size reflecting the fact that the precision of the sampleinformation increases as the number of samples increase. Theprobabilities of error were calculated using a normal distribu-tion model with a standard deviation of 10.60. This model wasfound to well describe counts of wasps caught in the flight testchamber. Additional details for this statistical model areprovided in the appendix.14. Interpretation of Results14.1 The quantity and quality of wasps in a shipment will beconsidered below the package claim when the test statistic issmaller than the selected critical value. If an acceptable errorrate is 0.05, then the CV for a sample size of 3 is 82 % (Table2). For the example presented in the Table 1, the test statistic(90 %) is larger than 82 %; therefore, the shipment would beclassified as meeting the package claim. If 5 samples wereFIG. 1 Test ChamberTABLE 1 Example—Based on 3 Random Samples and anExpectation of 60 Wasps/Card (the Package Claim)Sample NumberWasps on YellowCard157254351Average 162/3 = 54Observed/Expected · 100Observed/Expected ·100 = 90%Example: Test statistic = 90E2753 − 102used, the CV with an error rate of 0.05 is 86 % (Table 2)sotheshipment would again be classified as meeting the packageclaim. However, if 10 samples were used, the CV for an errorrate of 0.05 is approximately 91 %, so now the shipment wouldbe classified as not meeting the package claim.15. Precision and Bias15.1 The probabilities of error shown in Table 2 reflect theprecision of the test. The bias for E. eremicus has not beenassessed.16. Keywords16.1 Bemisia argentifolii; Bemisia tabaci; Eretmocerus er-emicus; Greenhouse Whitefly; purity; quantity; SilverleafWhitefly; Tobacco Whitefly; Trialeurodes vaporariorum;whitefly parasiteAPPENDIX(Nonmandatory Information)X1. STATISTICAL BACKGROUNDX1.1 The test assumes that a shipment meets the require-ment that the number of wasps that will emerge and fly is atleast 100 % of the package claim. Hence the null hypothesis is:Ho:µ $100 (X1.1)where:µ = the scaled mean number of wasps in a shipment that arecapable of flight.X1.2 We wish to determine a critical value (CV) with whichto compare a sample mean to so that the probability ofincorrectly declaring a shipment substandard is acceptable(e.g., ≤ 0.05). This CV will depend on the number of samplestaken from a shipment and will increase as the number ofsamples increase. Thus, we seek to find a CV such that:Pr$x¯ # CV ? µ $100% 5 0.05 (X1.2)X1.3 This is not possible unless µ is fixed. Because thisprobability will decrease for a fixed value of CV and increasingvalues of µ, we can set µ to 100 and determine CV so that:Pr$x¯ # CV ? µ 5 100% 5 0.05 (X1.3)X1.4 If sample counts (x) are distributed as normal randomvariables with known standard deviation, appropriate values ofCV can be determined using a normal cumulative distributionfunction.BIBLIOGRAPHY(1) van Lenteren , et al, “Guidelines for Quality Control of Commer-cially Produced Natural Enemies,” Quality Control and Productionof Biological Control Agents—Theory and Testing Procedures,J.C.van Lenteren, Ed., CABI Publishing, 2003, pp. 286–287.(2) Luczynski, A., et al, “Influence of Cold Storage on Pupal Develop-ment and Mortality During Storage and on Post-Storage Performanceof Encarsia formosa and Eretmocerus eremicus (Hymenoptera:Aphelinidae),” Biological Control, Vol 40, 2007, pp. 107–117.TABLE 2 Critical Values for the 3 Sample Sizes (3, 5 and 10Cards) and Corresponding Probabilities of ErrorProbability of Error for Sample Size of:Count as %of standard351084 0.00 0.00 0.0085 0.01 0.00 0.0086 0.01 0.00 0.0087 0.02 0.00 0.0088 0.02 0.01 0.0089 0.04 0.01 0.0090 0.05 0.02 0.0091 0.07 0.03 0.0092 0.10 0.05 0.0193 0.13 0.07 0.0294 0.16 0.10 0.0495 0.21 0.15 0.0796 0.26 0.20 0.12E2753 − 103ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. 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