1、Designation:D499489(Reapproved 2014)Standard Practice forRecovery of Viruses from Wastewater Sludges1This standard is issued under the fixed designation D4994;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.
2、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 practice is used for the recovery of viruses fromwastewater sludges and favors the enteroviruses.1.2 Both procedures are applicable
3、to raw,digested,anddewatered sludges.SectionsProcedure AAdsorption6 to 10Procedure BSonication11 to 151.3 This practice was tested on standardized sludges asdescribed in 10.1.It is the users responsibility to ensure thevalidity of this practice for untested matrices.1.4 The values stated in SI units
4、 are to be regarded asstandard.No other units of measurement are included in thisstandard.1.5 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices
5、and determine the applica-bility of regulatory limitations prior to use.1.6 Only adequately trained personnel should be allowed toperform these procedures and should use safety precautionsrecommended by the U.S.Public Health Service,Center forDisease Control,2for work with potentially hazardous biol
6、ogi-cal organisms.2.Referenced Documents2.1 ASTM Standards:3D1129 Terminology Relating to WaterD1193 Specification for Reagent Water3.Terminology3.1 DefinitionsFor definitions of terms used in thispractice,refer to Terminology D1129.4.Significance and Use4.1 Although many laboratories are presently
7、isolating vi-ruses from sludge,a valid comparison of data generated has notbeen possible because of the lack of a standard test method(s).5.Apparatus5.1 Centrifuge(s),refrigerated,capableofattaining10 000 g,screw-capped 100-mL centrifuge bottles that canwithstand 10 000 g,and 250-mL screw-capped cen
8、trifugebottles capable of withstanding 2 500 g.5.2 pH Meter,measuring to an accuracy of at least 0.1 pHunit,equipped with a combination-type electrode.Calibratewith standard buffers.5.3 Filter Apparatus,for membrane sterilization,4,5with47-mm diameter filter holder and 50-mL slip-tip syringe(see7.7
9、for type of filter material).6.Purity of Reagents6.1 Purity of ReagentsReagent grade chemicals shall beused in all tests.Unless otherwise indicated,it is intended thatall reagents shall conform to the specifications of the Commit-tee on Analytical Reagents of the American Chemical Society,where such
10、 specifications are available.6Other grades may beused,provided it is first ascertained that the reagent is ofsufficiently high purity to permit its use without lessening theaccuracy of the determination.1This practice is under the jurisdiction of ASTM Committee D19 on Water andis the direct respons
11、ibility of Subcommittee D19.24 on Water Microbiology.Current edition approved Jan.1,2014.Published March 2014.Originallyapproved in 1989.Last previous edition approved in 2009 as D4494 89(2009).DOI:10.1520/D4994-89R14.2Richardson,J.H.,and Barkley,W.E.,Biological Safety in Microbiological andBiomedic
12、al Laboratories,2nd edition,U.S.Dept.of Health and Human Services,Public Health Service,Center for Disease Control,and National Institutes of Healthand Human Services,1988.3For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual
13、 Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.4The sole source of supply of the apparatus,Swinnex filter(No.SX0047000),known to the committee at this time is Millipore Corp.,80Ashby Rd.,Bedford,MA01730.5If you are aware of alternative suppl
14、iers,please provide this information toASTM International Headquarters.Your comments will receive careful consider-ation at a meeting of the responsible technical committee,1which you may attend.6Reagent Chemicals,American Chemical Society Specifications,AmericanChemical Society,Washington,DC.For su
15、ggestions on the testing of reagents notlisted by the American Chemical Society,see Analar Standards for LaboratoryChemicals,BDH Ltd.,Poole,Dorset,U.K.,and the United States Pharmacopeiaand National Formulary,U.S.Pharmaceutical Convention,Inc.(USPC),Rockville,MD.Copyright ASTM International,100 Barr
16、 Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 6.2 Purity of WaterUnless otherwise indicated,referencesto water shall be understood to mean reagent water conformingto Specification D1193,Type II.PROCEDURE AADSORPTION7.Reagents and Materials7.1 Aluminum Chloride Solution(12.07 g/L)Dissolve12.07 g of aluminum chloride(AlCl36H2O)in 500 mL ofwater and dilute to 1000 mL.Autoclave AlCl3solution at121C for 15 min.7.2 Buffered Beef Extract SolutionDissolve 10 g of beefextract p
