ASTM D4994-19

Standard Practice for Recovery of Viruses from Wastewater Sludges

SIGNIFICANCE AND USE
4.1 Although many laboratories are presently isolating viruses from sludge, a valid comparison of data generated has not been possible because of the lack of a standard test method(s).
SCOPE
1.1 This practice is used for the recovery of viruses from wastewater sludges and favors the enteroviruses.  
1.2 Both procedures are applicable to raw, digested, and dewatered sludges.    
Sections  
Procedure A—Adsorption  
7 to 10  
Procedure B—Sonication  
11 to 15  
1.3 This practice was tested on standardized sludges as described in 10.1. It is the user's responsibility to ensure the validity of this practice for untested matrices.  
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 Only adequately trained personnel should be allowed to perform these procedures and should use safety precautions recommended by the U.S. Public Health Service, Center for Disease Control,2  for work with potentially hazardous biological organisms.  
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.

General Information

• Status: Published
• Publication Date: 31-Mar-2019
• ICS: 13.060.30 - Sewage water
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Standards Content (Sample)

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.
Designation: D4994 − 19
Standard Practice for
1
Recovery of Viruses from Wastewater Sludges
This standard is issued under the fixed designation D4994; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope* 2. Referenced Documents
3
2.1 ASTM Standards:
1.1 This practice is used for the recovery of viruses from
D1129Terminology Relating to Water
wastewater sludges and favors the enteroviruses.
D1193Specification for Reagent Water
1.2 Both procedures are applicable to raw, digested, and
dewatered sludges.
3. Terminology
Sections
3.1 Definitions:
3.1.1 For definitions of terms used in this standard, refer to
Procedure A—Adsorption 7 to 10
Procedure B—Sonication 11 to 15 Terminology D1129.
1.3 This practice was tested on standardized sludges as
4. Significance and Use
described in 10.1. It is the user’s responsibility to ensure the
4.1 Although many laboratories are presently isolating vi-
validity of this practice for untested matrices.
rusesfromsludge,avalidcomparisonofdatageneratedhasnot
1.4 The values stated in SI units are to be regarded as
been possible because of the lack of a standard test method(s).
standard. No other units of measurement are included in this
standard. 5. Apparatus
5.1 Centrifuge(s), refrigerated, capable of attaining
1.5 This standard does not purport to address all of the
10000× g, screw-capped 100-mL centrifuge bottles that can
safety concerns, if any, associated with its use. It is the
withstand 10000× g, and 250-mL screw-capped centrifuge
responsibility of the user of this standard to establish appro-
bottles capable of withstanding 2500× g.
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use. 5.2 pH Meter, measuring to an accuracy of at least 0.1 pH
unit, equipped with a combination-type electrode. Calibrate
1.6 Only adequately trained personnel should be allowed to
with standard buffers.
perform these procedures and should use safety precautions
4,5
recommended by the U.S. Public Health Service, Center for 5.3 Filter Apparatus, for membrane sterilization, with
2
47-mm diameter filter holder and 50-mL slip-tip syringe (see
Disease Control, for work with potentially hazardous biologi-
7.7 for type of filter material).
cal organisms.
1.7 This international standard was developed in accor-
6. Purity of Reagents
dance with internationally recognized principles on standard-
6.1 Purity of Reagents—Reagent grade chemicals shall be
ization established in the Decision on Principles for the
used in all tests. Unless otherwise indicated, it is intended that
Development of International Standards, Guides and Recom-
all reagents shall conform to the specifications of the Commit-
mendations issued by the World Trade Organization Technical
tee onAnalytical Reagents of theAmerican Chemical Society,
Barriers to Trade (TBT) Committee.
3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
This practice is under the jurisdiction ofASTM Committee D19 on Water and contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
is the direct responsibility of Subcommittee D19.24 on Water Microbiology. Standards volume information, refer to the standard’s Document Summary page on
Current edition approved April 1, 2019. Published April 2019. Originally the ASTM website.
4
approved in 1989. Last previous edition approved in 2014 as D4494–89 (2014). The sole source of supply of the apparatus, Swinnex filter (No. SX0047000),
DOI: 10.1520/D4994-19. knowntothecommitteeatthistimeisMilliporeCorp.,80AshbyRd.,Bedford,MA
2
Richardson,J.H.,andBarkley,W.E., Biological Safety in Microbiological and 01730.
5
Biomedical Laboratories, 2nd edition, U.S. Dept. of Health and Human Services, If you are aware of alternative suppliers, please provide this information to
PublicHealthService,CenterforDiseaseControl,andNationalInstitutesofHealth ASTM International Headquarters. Your comments will receive careful consider-
1
and Human Services, 1988. ation at a meeting of the responsible technical committee, which you may attend.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

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D4994 − 19
6
where such specifications are available. Other grades may be means and further concentrated by
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: D4994 − 89 (Reapproved 2014) D4994 − 19
Standard Practice for
1
Recovery of Viruses from Wastewater Sludges
This standard is issued under the fixed designation D4994; 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 Scope*
1.1 This practice is used for the recovery of viruses from wastewater sludges and favors the enteroviruses.
1.2 Both procedures are applicable to raw, digested, and dewatered sludges.
Sections
Procedure A—Adsorption 6 to 10
Procedure A—Adsorption 7 to 10
Procedure B—Sonication 11 to 15
1.3 This practice was tested on standardized sludges as described in 10.1. It is the user’s responsibility to ensure the validity
of this practice for untested matrices.
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 safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.6 Only adequately trained personnel should be allowed to perform these procedures and should use safety precautions
2
recommended by the U.S. Public Health Service, Center for Disease Control, for work with potentially hazardous biological
organisms.
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.
2. Referenced Documents
3
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
3. Terminology
3.1 Definitions—Definitions: For definitions of terms used in this practice, refer to Terminology D1129.
3.1.1 For definitions of terms used in this standard, refer to Terminology D1129.
4. Significance and Use
4.1 Although many laboratories are presently isolating viruses from sludge, a valid comparison of data generated has not been
possible because of the lack of a standard test method(s).
5. Apparatus
5.1 Centrifuge(s), refrigerated, capable of attaining 10 000 × g, screw-capped 100-mL centrifuge bottles that can withstand
10 000 × g, and 250-mL screw-capped centrifuge bottles capable of withstanding 2 500 × 2500 × g.
1
This practice is under the jurisdiction of ASTM Committee D19 on Water and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved Jan. 1, 2014April 1, 2019. Published March 2014April 2019. Originally approved in 1989. Last previous edition approved in 20092014 as
D4494 – 89 (2009).(2014). DOI: 10.1520/D4994-89R14.10.1520/D4994-19.
2
Richardson, J. H., and Barkley, W. E., Biological Safety in Microbiological and Biomedical Laboratories, 2nd edition, U.S. Dept. of Health and Human Services, Public
Health Service, Center for Disease Control, and National Institutes of Health and Human Services, 1988.
3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D4994 − 19
5.2 pH Meter, measuring to an accuracy of at least 0.1 pH unit, equipped with a combination-type electrode. Calibrate with
standard buffers.
4,5
5.3 Filter Apparatus, for membrane sterilization, with 47-mm diameter filter holder and 50-mL slip-tip syringe (see 7.7 for
type of filter material).
6. Purity of Reagents
6.1 Purity of Reagents—Reagent grade chemicals shall be used in all tests. Unless otherwise indicated, it is intended that all
reagents shall conform to the specifications of the Committee on Analytical Reagents of the American Chemical Society, where
6
such specifications are available. Other grades may be used, provided it is first ascertained that the reagen
...