1、 Collection of SANS standards in electronic format (PDF) 1. Copyright This standard is available to staff members of companies that have subscribed to the complete collection of SANS standards in accordance with a formal copyright agreement. This document may reside on a CENTRAL FILE SERVER or INTRA
2、NET SYSTEM only. Unless specific permission has been granted, this document MAY NOT be sent or given to staff members from other companies or organizations. Doing so would constitute a VIOLATION of SABS copyright rules. 2. Indemnity The South African Bureau of Standards accepts no liability for any
3、damage whatsoever than may result from the use of this material or the information contain therein, irrespective of the cause and quantum thereof. ISBN 978-0-626-22313-7 SANS 5277:2009Edition 4SOUTH AFRICAN NATIONAL STANDARD Resistance to attack by Chaetomium globosum Published by SABS Standards Div
4、ision 1 Dr Lategan Road Groenkloof Private Bag X191 Pretoria 0001Tel: +27 12 428 7911 Fax: +27 12 344 1568 www.sabs.co.za SABS SANS 5277:2009 Edition 4 Table of changes Change No. Date Scope Foreword This South African standard was approved by National Committee SABS SC 1028G, Pesticides Resistance
5、to fungal attack, in accordance with procedures of the SABS Standards Division, in compliance with annex 3 of the WTO/TBT agreement. This document was published in June 2009. This document supersedes SABS SM 277:1975 (third edition). SANS 5277:2009 Edition 4 1 Contents Page Foreword 1 Scope 3 2 Norm
6、ative references . 3 3 Apparatus and materials . 3 4 Test fungus and culture medium 3 5 Maintenance of the test fungus. 4 6 Preparation of test specimens 4 7 Procedure 4 8 Evaluation 5 SANS 5277:2009 Edition 4 2 This page is intentionally left blank SANS 5277:2009 Edition 4 3 Resistance to attack by
7、 Chaetomium globosum 1 Scope This standard specifies a method for the determination of the resistance of the test specimens to attack by Chaetomium globosum. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only t
8、he edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. Information on currently valid national and international standards can be obtained from the SABS Standards Division. SANS 171, Glassware and equipment for microbiologi
9、cal tests. 3 Apparatus and materials 3.1 Apparatus The apparatus and equipment shall comply with the requirements of SANS 171. 3.2 Materials 3.2.1 Distilled water, that has been distilled in an all-glass apparatus or de-mineralized water of equivalent purity. 3.2.2 Sterile distilled water. 3.2.3 A n
10、on-ionic wetting agent, that is non-toxic to the test fungus. 4 Test fungus and culture medium 4.1 Test fungus Chaetomium globosum, ATCC 16021, SABS TCC 15 or ATCC 36703, SABS TCC 381. SANS 5277:2009 Edition 4 4 4.2 Culture medium for the test fungus 4.2.1 Weigh out accurately: Ammonium nitrate (NH4
11、NO3)3,0 g Mono-potassium phosphate (KH2PO4).2,5 g Di-potassium phosphate (K2HPO4) 2,0 g Magnesium sulfate, heptahydrate (MgSO4 7H2O) .0,2 g Ferrous sulfate, heptahydrate (FeSO4 7H2O) 0,1 g 4.2.2 Dissolve the salts in approximately 800 mL of distilled water. Add 20,0 g of agar and boil to dissolve. 4
12、.2.3 Allow to cool to approximately 60 C and by adding either 0,1 M sodium hydroxide solution or 0,1 M hydrochloric acid, as might be required, so adjust the pH value of the medium that it will be 6,3 0,2 after sterilization. 4.2.4 Add distilled water to bring the volume to 1 L. Dispense into suitab
13、le containers for storage and into test tubes for the cultivation of the test fungus. Sterilize by autoclaving for 20 min at 121 C 2 C. 4.2.5 Allow the test tubes to cool in a slanted position that provides maximum growth surface. 5 Maintenance of the test fungus Subculture the test fungus at interv
14、als of 14 d on sterile strips (approximately 25 mm 12 mm) of filter paper placed on slants of the culture medium in suitable tubes. 6 Preparation of test specimens 6.1 When relevant, subject the test specimens to the appropriate pre-treatment specified in the relevant standard for the required bacte
15、ria. 6.2 The quantity and dimensions of the test specimens shall be as specified in the relevant standard for the required bacteria. 7 Procedure 7.1 Sterilization of test specimens 7.1.1 Place the test specimens in a suitable container and add distilled water that contains 0,05 % of the non-ionic we
16、tting agent until they are completely covered. If the specimens are to be used subsequently for testing of water-repellency, then omit the wetting agent. 7.1.2 Keep the specimens submerged for 1 h, then pour off the solution, close the container, and sterilize by autoclaving for 10 min at 121 C 2 C.
17、 7.2 Spore suspension 7.2.1 Add 10 mL of the sterile distilled water that contains 0,05 % of the non-ionic wetting agent to a slant culture (see clause 5) of the fungus. 7.2.2 Using a sterile inoculating wire, gently scrape the surface growth from the culture of the test fungus. SANS 5277:2009 Editi
18、on 4 5 7.2.3 Filter the spore suspension through sterile coarse cotton gauze into a sterile flask that contains 50 mL of the sterile distilled water and shake the flask. 7.3 Inoculation 7.3.1 Using sterile forceps, aseptically transfer each test specimen to a Petri dish that contains a depth of 4 mm
19、 to 5 mm of the sterile culture medium. Ensure that one surface of each specimen is in firm contact with the culture medium and that each end is bent up vertically (away from the agar surface) for a distance of at least 25 mm. NOTE In cases where a breaking strength or other test is to be used for s
20、ubsequent evaluation, it is advisable not to permit areas of the test specimens that are to be inserted later in the jaws of a tensile strength testing machine (or used for similar anchoring purposes) to come into contact with the agar surface. 7.3.2 Place a sterile strip (approximately 25 mm 12 mm)
21、 of filter paper on the culture medium but not in contact with the test specimen. 7.3.3 Spray the spore suspension (see 7.2) on to the surface of each test specimen and on to the surface of each strip of filter paper to the point of run-off. Ensure that the spore suspension is evenly distributed ove
22、r the surface of each test specimen. 7.3.4 The room or space where the spraying takes place shall be frequently disinfected. 7.4 Incubation Incubate the Petri dishes at 28 C 2 C and a relative humidity of 90 % 5 % for a period of 14 d. 7.5 Visual examination 7.5.1 After incubation, note whether the
23、surface of each filter paper strip in the Petri dishes is covered with the fruiting bodies of the fungus. Remove the test specimens from the Petri dishes and examine them for fungal growth. 7.5.2 In cases where fungal growth is not evident or where there are signs of contamination, discard the speci
24、mens and repeat the test on a new set of test specimens. 8 Evaluation 8.1 Visual examination Report the extent of fungal growth on the control filter paper and on the test specimens. 8.2 Other tests Perform any other tests in accordance with the method(s) specified in the relevant standard for the r
25、equired bacteria. SABS This page has been left blank intentionally SABS Standards Division The objective of the SABS Standards Division is to develop, promote and maintain South African National Standards. This objective is incorporated in the Standards Act, 2008 (Act No. 8 of 2008). Amendments and
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