ASTM D4994-89(1996)

Standard Practice for Recovery of Viruses from Wastewater Sludges

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 B---Sonication 11 to 15
1.3 This practice was tested on standardized sludges as described in 10.1 and 17.1. It is the user's responsibility to ensure the validity of this practice for untested matrices.
1.4 This standard may involve hazardous materials, operations, and equipment. This standard does not purport 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 and health practices and determine the applicability of regulatory limitations prior to use.  
1.5 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,   for work with potentially hazardous biological organisms.

General Information

• Status: Historical
• Publication Date: 31-Dec-1995
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Relations

• Replaced By: ASTM D4994-89(2002) - Standard Practice for Recovery of Viruses from Wastewater Sludges
• Effective Date: 27-Oct-1989

Standards Content (Sample)

Designation: D 4994 – 89 (Reapproved 1996)
AMERICAN SOCIETY FOR TESTING AND MATERIALS
100 Barr Harbor Dr., West Conshohocken, PA 19428
Reprinted from the Annual Book of ASTM Standards. Copyright ASTM
Standard Practice for
1
Recovery of Viruses from Wastewater Sludges
This standard is issued under the fixed designation D 4994; 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 10 000 3 g, screw-capped 100-mL centrifuge bottles that can
withstand 10 000 3 g, and 250-mL screw-capped centrifuge
1.1 This practice is used for the recovery of viruses from
bottles capable of withstanding 2 500 3 g.
wastewater sludges and favors the enteroviruses.
5.2 pH Meter, measuring to an accuracy of at least 0.1 pH
1.2 Both procedures are applicable to raw, digested, and
unit, equipped with a combination-type electrode. Calibrate
dewatered sludges.
with standard buffers.
Sections
4
5.3 Filter Apparatus, for membrane sterilization, with
Procedure A—Adsorption 6 to 10
Procedure B—Sonication 11 to 15
47-mm diameter filter holder and 50-mL slip-tip syringe (see
7.7 for type of filter material).
1.3 This practice was tested on standardized sludges as
described in 10.1 and 17.1. It is the user’s responsibility to
6. Purity of Reagents
ensure the validity of this practice for untested matrices.
6.1 Purity of Reagents—Reagent grade chemicals shall be
1.4 This standard does not purport to address all of the
used in all tests. Unless otherwise indicated, it is intended that
safety concerns, if any, associated with its use. It is the
all reagents shall conform to the specifications of the Commit-
responsibility of the user of this standard to establish appro-
tee on Analytical Reagents of the American Chemical Society,
priate safety and health practices and determine the applica-
5
where such specifications are available. Other grades may be
bility of regulatory limitations prior to use.
used, provided it is first ascertained that the reagent is of
1.5 Only adequately trained personnel should be allowed to
sufficiently high purity to permit its use without lessening the
perform these procedures and should use safety precautions
accuracy of the determination.
recommended by the U.S. Public Health Service, Center for
2
6.2 Purity of Water—Unless otherwise indicated, references
Disease Control, for work with potentially hazardous biologi-
to water shall be understood to mean reagent water conforming
cal organisms.
to Specification D 1193, Type II.
2. Referenced Documents
PROCEDURE A—ADSORPTION
2.1 ASTM Standards:
3
D 1129 Terminology Relating to Water
3
7. Reagents and Materials
D 1193 Specification for Reagent Water
7.1 Aluminum Chloride Solution (12.07 g/L)—Dissolve
3. Terminology
12.07 g of aluminum chloride (AlCl ·6H O) in 500 mL of
3 2
3.1 Definitions—For definitions of terms used in this prac-
water and dilute to 1000 mL. Autoclave AlCl solution at
3
tice, refer to Terminology D 1129.
121°C for 15 min.
7.2 Buffered Beef Extract Solution—Dissolve 10 g of beef
4. Significance and Use
6
extract powder, 1.34gofNa HPO ·7H O, and 0.12 g of citric
2 4 2
4.1 Although many laboratories are presently isolating vi-
acid in 100 mL of water in a screw-cap flask by stirring for
ruses from sludge, a valid comparison of data generated has not
about2hona magnetic stirrer. Autoclave at 121°C for 15 min.
been possible because of the lack of a standard test method(s).
7.3 Disodium Hydrogen Phosphate Solution (4 g/100
mL)—Dissolve4gof disodium hydrogen phosphate
5. Apparatus
5.1 Centrifuge(s), refrigerated, capable of attaining
4
The Swinnex filter (No. SX0047000, available from Millipore Corp., 80 Ashby
Rd., Bedford, MA 01730, or equivalent, has been found suitable for this purpose.
5
1
Reagent Chemicals, American Chemical Society Specifications, American
This practice is under the jurisdiction of ASTM Committee D-19 on Water and
Chemical Society, Washington, DC. For suggestions on the testing of reagents not
is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
listed by the American Chemical Society, see Analar Standards for Laboratory
Current edition approved Oct. 27, 1989. Published March 1990.
2
Richardson, J. H., and Barkley, W. E., Biological Safety in Microbiological and Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
Biomedical Laboratories, 2nd. edition, U.S. Dept. of Health and Human Services, and National Formulary, U.S. Pharmaceutical Convention, Inc. (USPC), Rockville,
Public Health Service, Center for Disease Control, and National Institutes of Health MD.
6
and Human Services, 1988. Extract available from Grand Island Biological Corp., 3175 Staley Rd, Grand
3
Annual Book of ASTM Standards, Vol 11.01. Island, NY 14072, or equivalent, has been found
...