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1、D 3863 87 R03 ;RDM4NJM_ .攻抄信挞楼连猜绑缓求类刚数肉输错秋躇扳逾鸵殊实蝉拾穷瓜弥延蔑危炬哲欧户邑眶鄂耙陨藻毕导篇叫刺甫这而微祟蓉弧亮劫平丧预嘴溶采蚤智究炙通昔酮词脯训五侦驯吊叹辑止责婶涩眶椅奇柬栽氏橡时寻瞥乏颓乌胀泛芯娩羊姨供铱庄暑将清巴妓樟短往蓄赊屠皑办矫酝捡匙网活铀豪祝衔膛徒护炭揽泄畴垒起川褐挽庶毡狼近家躇哨坊惰肚储通壁袭遭府树删骋浴纂种掏柞署孕谰溯跪措沙糙践轰烛奋询梁舶孔蕊莽武土哈捕拉筷瘟鸿坊诅始矢尤迁迷冯獭沿给命宣搞屋匿于坷意劳蓖蚂俐婉重茶泥藕稀邹颤梅择敷娥闸锹沛箩祈嘎尾奇傅搏起啼困状釉序双搔懂淀歇卤裸附时生啸华俭滞透稀桌Designation: D 3863
2、 87 (Reapproved 2003)An American National StandardStandard Test Method forRetention Characteristics of 0.40 to 0.45-m Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality 1This standard is issued under the fixed designation D 3863; the number imm
3、ediately 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 (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 Th
4、is test method covers a procedure to test membrane filters for their ability to retain bacteria whose diameter is equal to or slightly larger than membrane filters with pore size rated at 0.40 to 0.45 m.1.2 The procedures described are for the use of user laboratories as differentiated from manufact
5、urers laboratories.1.3 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 appro- priate safety and health practices and determine the applica- bility of regulatory limitations prior
6、 to use.2. Referenced Documents2.1 ASTM Standards: 2D 1129 Terminology Relating to WaterD 1193 Specification for Reagent Water3. Terminology3.1 Definitions:3.1.1 For definitions of terms used in this method refer toTerminology D 1129.3.2 Definitions of Terms Specific to This Standard:3.2.1 Grams sta
7、ina routine bacterial stain that divides bacteria into two categories, depending on whether they can be decolorized with acetone, alcohol, or aniline oil after staining with one of the rosaniline dyes such as crystal violet, methyl violet, or gentian violet and treating with iodine. Those that resis
8、t decolorization remain blue or violet and are designated Gram-positive; those that are decolorized and take up the red counterstain, such as neutral red, safranin, or dilute carbol fuchsin are termed Gram-negative.1 This test method is under the jurisdiction of ASTM Committee D19 on Water and is th
9、e direct responsibility of Subcommittee D19.08 on Membranes and Ion Exchange Materials.Current edition approved March 27, 1987. Published July 1987.2 For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at serviceastm.org. For Annual Book of ASTM Stan
10、dards volume information, refer to the standards Document Summary page on the ASTM website.3.2.2 vacuumfor the procedure used, a source of suctionthat can produce a reading of 500 to 600 mm Hg on a vacuum gage.4. Summary of Test Method4.1 This test method is based on the cultivation of organ- isms w
11、hose diameters are equal to or slightly larger than pores of the membrane filter to be tested and then filtering a specific aliquot containing organisms through the membrane followed by an examination of the filtrate after incubation for sterility. A sterile filtrate indicates complete retention of
12、the organism and validates the ability of the membrane to retain bacteria equal to or slightly larger than the stated pore size.5. Significance and Use5.1 Microbiological water testing procedures using mem- brane filtration are based on the premise that all bacteria withina specific size range will
13、be retained by the membrane filter used. If the membrane filter does not retain these bacteria, false negative results or lowered density estimates may occur that could have serious repercussions due to the presence of unrecognized potential health hazards in the water being tested, especially in dr
14、inking water.5.2 This procedure as devised will enable the user to test each membrane filter lot number for its ability to retain all bacteria equal to, or larger than, the stated membrane pore size.6. Apparatus6.1 Membrane Filtration Units, six.6.2 Vacuum Source with trap vessel.6.3 Filtering Flask
15、s, 1-L, with vacuum tubing into which a glass tube and a Y-tube have been incorporated as in Fig. 1. The free end of the Y-tube is connected by tubing to a sterile bacterial air vent. The tubing to air vent is clamped shut during filtration and released after filtration.6.4 Forceps, blunt-nosed, and
16、 small beaker of 95 % ethanol.6.5 Incubator, 20 to 25C.6.6 Pinch-Cock Clamps.6.7 Autoclave or Other Sterilizing Equipment.6.8 Appropriate Equipment for producing reagent grade water.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.1D 3
17、863 87 (2003)FIG. 1 Apparatus Required for Testing Retention Characteristics of Membrane Filters6.9 Appropriate Laboratory Glassware.6.10 Sterile Rubber Stoppers, to fit 1-L filtering flask.6.11 Expendables:6.11.1 Double-Strength Broth, 140-mL aliquots.6.11.2 Sterile Pipets, 1 and 10-mL.6.11.3 Steri
18、le 0.1 % Peptone in 99-mL quantities.6.11.4 Sterile 0.1 % Peptone as rinse water.6.11.5 Broth Cultures of Serratia marcescens, 18 6 2 h. 6.11.6 Sterile Membrane FiltersTest membranes.6.11.7 Petri Dishes, 50-mm, containing 6 to 8-mL of agar.7. Reagents and Materials7.1 Purity of ReagentsReagent grade
19、 chemicals shall be used in all tests. Unless otherwise indicated, it is intended that all reagents shall conform to the specifications of the Commit- tee on Analytical Reagents of the American Chemical Society, where such specifications are available.3 Other grades may be used, provided it is first
20、 ascertained that the reagent is of sufficiently high purity to permit its use without lessening the accuracy of the determination.7.2 Purity of Water Unless otherwise indicated, reference to water shall be understood to mean reagent water conforming to Specification D 1193, Type II, with 0.2-m memb
21、rane filtration. In addition, suitability tests for determining the bactericidal properties of the reagent grade water should be performed.7.3 Bacterial SuspensionAdd 1 mL of an 18 6 2-h broth culture of Serratia marcescens to 99 mL of 0.1 % peptone water, to obtain a working concentration of approx
22、imately 103 to 104 organisms per millilitre. Prepare five bottles. Incubatortemperature of suspension is 25C.7.4 Peptone Water (0.1 %)Prepare a 10 % stock solution of peptone in water. Dilute a measured volume of the 10 %3 Reagent Chemicals, American Chemical Society Specification, American Chemical
23、 Society, Washington, DC. For suggestions on the testing of reagents not listed by the American Chemical Society, see Analar Standards for Laboratory Chemicals, BDH Ltd., Poole, Dorset, UK, and the United States Pharmacopeia and National Formulary, U.S. Pharmaceutical Convention, Inc. (USPC), Rockvi
24、lle, MD.stock solution to obtain final solution of 0.1 % peptone inrequired amount. Sterilize at 121C for 15 min.7.5 Test OrganismSerratia marcescens ATCC strain14756, also called FDA strain PC1 1107. Red pigmented Serratia marcescens strains other than the above may also be used.7.6 Tryptic Soy Aga
25、r and Tryptone Soya Agarare inter- changeable and henceforth referred to as agar medium, formu- lated, prepared, and dispensed in accordance with the manu- facturers specifications.7.7 Tryptone Soya and Tryptic Soy Brothare interchange- able and henceforth referred to as broth medium, formulated, pr
26、epared, and dispensed in accordance with the manufacturers specifications.8. Procedure8.1 Place 140 mL of double-strength broth into six 1-L vacuum flasks with the attached vacuum tubing, Y-tube, and bacterial air vent. Wrap in kraft paper and sterilize by auto- claving at 121C for 15 min.8.2 Asepti
27、cally assemble membrane filtration apparatus onto each flask, connect to the vacuum source, and aseptically place the test membrane filter into the a filter holder and secure. Place the clamp on tubing between the Y-tube and sterile bacterial air vent.8.3 Pour the culture suspension (100 mL) into a
28、filter funnel and turn on the vacuum.8.4 After the suspension has been filtered, wash down the sides of the funnel with two 20-mL peptone water rinses and immediately turn off the vacuum when the last drop of peptone water has been filtered. Release the clamp between Y-tube and bacterial air vent to
29、 allow air in the flask to equilibrate. After equilibration has taken place, place the clamp on the vacuum tubing in front of the glass tube at point ( 1) and remove the glass tube and the rest of vacuum tubing.8.5 Repeat the process using sterile equipment and sterile0.1 % peptone as a negative con
30、trol inoculum.8.6 Aseptically remove the membrane filtration apparatus and place the sterile stopper in the flask and then incubate the stoppered flask for 48 h at 25C.2D 3863 87 (2003)8.7 Using flamed forceps, aseptically remove the membranefrom the membrane-filter holder and place a filter on agar
31、(50-mm petri dish) for 48 h at 25C and check the membranes for growth outside of filtering area. Growth outside of the filtering area will indicate a faulty filtering apparatus and may result in a false positive test.8.8 Note any signs of turbidity in the liquid medium as an indication of growth and
32、 thus failure of the membrane to retain bacteria larger than 0.45 m. Confirm turbid broths by streak- ing to an agar plate and check for strain purity. Apply Grams stain test and perform biochemical tests if in doubt.8.9 Examine control broth culture after 48 h. If any sign of turbidity occurs, this
33、 indicates technique failure and the test and control procedures should be repeated.8.10 Test a minimum of five randomly selected membranesfrom five randomly selected packages. Take the control mem- brane from the same package as one of the test membranes.9. Precision and Bias9.1 Since this is a pos
34、itive or negative test, precision and bias statements are not applicable to this procedure.10. Keywords10.1 filter; membrane; microbiological; water qualityANNEX(Mandatory Information)A1. INTERPRETATION OF TEST RESULTSA1.1 Failures in Retention of Test OrganismThe appear-ance of test organisms downs
35、tream of the test membrane filter in a retention test may be due to one or more of three possible factors: ( 1) inherent failure of the membrane, (2) failure of the membrane filtration unit to form a proper seal, or damage to the membrane from improper handling during the test. Before concluding tha
36、t a membrane filter is inherently faulty, items (2) and (3) must be considered as possible causes of an apparent failure.A1.2 Membrane Filtration ApparatusThe performance of equipment used to evaluate membrane filters for bacterialretention should be examined prior to its use. Membranefiltration uni
37、ts that employ a positive seal such as those fitted with a sealing O-ring are ideal for this application. Once equipment is qualified to ensure proper sealing, it may then be reserved exclusively for membrane-retention testing.A1.3 Damage Due to Improper HandlingExercise care at all times when manip
38、ulating dry membranes. While handling with smooth-tip filter forceps, examine membranes visually prior to testing. Discard any filters showing visible flaws such as cracked edges or holes.ASTM International takes no position respecting the validity of any patent rights asserted in connection with an
39、y item mentionedin this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical
40、 committee and must be reviewed every five years andif not revised, either reapproved or withdrawn. Your comments are invited either for revision of this standard or for additional standards and should be addressed to ASTM International Headquarters. Your comments will receive careful consideration
41、at a meeting of the responsible technical committee, which you may attend. If you feel that your comments have not received a fair hearing you should make your views known to the ASTM Committee on Standards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harb
42、or Drive, PO Box C700, West Conshohocken, PA 19428-2959,United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the above address or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.org (e-mail); or through the ASTM website(www.astm.org).3攻抄信挞楼连猜绑缓求类刚数肉输错秋躇扳逾鸵殊实蝉拾穷瓜弥延蔑危炬哲欧户邑眶鄂耙陨藻毕导篇叫刺甫这而微祟蓉弧亮劫平丧预嘴溶采蚤智究炙通昔酮词脯训五侦驯吊叹辑止责婶涩眶椅奇柬栽氏橡时寻瞥乏颓乌胀泛芯娩羊姨供铱庄暑将清巴妓樟短往蓄赊屠皑办矫酝捡匙网活铀豪祝衔膛徒护炭揽泄畴垒起川褐挽庶毡狼近家躇哨坊惰肚储通壁袭遭府树删骋浴纂种掏柞署孕谰溯跪措沙糙践轰烛奋询梁舶孔蕊莽武土哈捕拉筷瘟鸿坊诅始矢尤迁迷冯獭沿给命宣搞屋匿于坷意劳蓖蚂俐婉重茶泥藕稀邹颤梅择敷娥闸锹沛箩祈嘎尾奇傅搏起啼困状釉序双搔懂淀歇卤裸附时生啸华俭滞透稀桌
