MFLP-60 Screening Bottled Water (Water in Sealed Containers) for the Presence of Indicator and Pathogenic Bacteria
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48F1F3981E2043FEACE41C4CB4EF8BAF |
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0.02 |
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3 |
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2024-7-30 |
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Published by: POLYSCIENCE PUBLICATIONS, P.O. Box 1606, Station St-Martin, Laval, Quebec,Canada H7V 3P9. TEL.: 1-800-840-5870. FAX: (450) 688-1930.,Government of Canada Gouvernement du Canada,Laboratory Procedure MFLP-60,September 1999,HEALTH PROTECTION BRANCH,OTTAWA,SCREENING BOTTLED WATER (WATER IN SEALED CONTAINERS) FOR,THE PRESENCE OF INDICATOR AND PATHOGENIC BACTERIA.,Don Warburton,Bureau of Microbial Hazards, Food Directorate,Postal Locator: 2204A1,Health Canada, Ottawa ON K1A 0L2,E-mail: Don_Warburton@hc-sc.gc.ca,1. APPLICATION,This method is applicable to the detection of viable bacteria in bottled water (water in sealed containers;,including mineral and spring water) to determine compliance with the requirements of Sections 4 and 7,of the Food and Drugs Act and Division 12 of the Regulations.,2. PRINCIPLE,This screening method is based on the method of Warburton (7.1 ) for detecting viable microorganisms,in bottled water that has been treated by a disinfection step (e.g. ozonation, micro-filtration, UV light or,other means) and/or is carbonated. After determining the presence of aerobic colony counts (ACC),and/or growth in Presence Absence Broth and Buffered Peptone Water, selective and differential agars,can be used to aid in isolation and identification of coliforms, Aeromonas hydrophila, Escherichia coli,Pseudomonas aeruginosa and other bacteria, following procedures published in this Compendium.,3. DEFINITION OF TERMS,See Appendix A of Volume 3.,4. COLLECTION OF SAMPLES,4.1 See Appendix B of Volume 3.,4.2 Each sample unit shall contain at least 500 mL.,4.3 Keep the sample units refrigerated (0-4oC) or frozen during transport, depending on the nature,of the product.,5. MATERIALS AND SPECIAL EQUIPMENT,1) HGMF (1600 grid-cell, 0.45μm pore size) (ISO-GRIDR; available from Oxoid Inc., Ottawa, ON) or,equivalent.,2) Membrane filter forceps (Millipore Corp.),3) Tryptic soy agar (TSA; commercially available),MFLP-60,- 2 - September 1999,4) Buffered Peptone Water (BPW; commercially available),5) Presence Absence Broth (PA; commercially available); make single strength (1X).,Note: preparation instructions give triple strength (3X) for waste water analysis.,NOTE: It is the reponsibility of each laboratory to ensure that the temperature of the incubators or waterbaths,are maintained at the recommended temperatures. Where 35EC is recommended in text of the,method the incubator may be at 36 +/-1.0E C. Similarly, lower temperatures of 30 or,25 may be +/- 1.0EC. However, where higher temperatures are recommended, such as 43 or 45.5EC,it is imperative that the incubators or waterbaths be maintained within 0.5EC due to potential lethality,of higher temperatures on the microorganism being isolated.,6. PROCEDURE,Samples may be analyzed individually or after compositing. Carry out the test in accordance with the,following instructions:,6.1 Handling of Sample Units,6.1.1 In the laboratory prior to analysis keep sample units refrigerated (0-4EC).,6.1.2 Analyze sample units as soon as possible after receipt in the laboratory.,6.2 Preparation for Analysis,6.2.1 Have ready sterile tryptic soy agar (TSA) plates or flasks of 225 mL single strength (1X),presence absence broth or buffered peptone water.,6.2.2 Clean the surface of the working area with a suitable disinfectant. Continue with 6.3 or 6.4.,6.3 Filtration,6.3.1 Clearly label duplicate Petri dishes with appropriate identifiying information.,6.3.2 The HGMF will allow counts to be made from suspensions containing up to 5,000,organisms/mL. There normally should be no need to prepare further dilutions.,6.3.3 Agitate each sample to resuspend material that may have settled out.,6.3.4 Handle HGMF with sterile membrane filter forceps.,6.3.5 Following the manufacturer's instructions for use of the filtration apparatus, pour 100 mL of the,analytical unit into it. Open the filter valve until all liquid has passed through and aseptically,remove the HGMF. Do in duplicate.,6.3.6 Transfer the HGMF to the surface of a TSA plate, rolling it onto the agar to avoid trapping air,bubbles. Incubate plates right side up in stacks of not more than three, incubate at 35EC for,18-24 h .,6.3.7 Follow the manufacturer's instructions for cleaning the filtration apparatus.,6.3.8 If HGMF's do not contain growth, record test results as <1/100 mL.,6.3.9 Count and record results. Continue as below in 6.5.,MFLP-60,- 3 - September 1999,6.4 Inoculation of Broths,6.4.1 Aseptically transfer 100 mL of the sample unit to either PA (1X) or BPW. Incubate broths at,35EC for 18-24 h.,6.4.2 After incubation, PA broth will be yellow in colour and contain gas if coliforms are pre……
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