3 Interpretation Guide Petrifilm Aerobic Count Plate This guide familiarizes you with results on 3M Petrifilm Aerobic Count Plates. For more information, contact the official 3M Microbiology Products representative nearest you. The Petrifilm Aerobic Count (AC) plate is a ready-made culture medium system that contains Standard Methods nutrients, a cold-water-soluble gelling agent, and an indicator that facilitates colony enumeration. Petrifilm AC plates are used for the enumeration of aerobic bacteria. Aerobic Bacteria Count = 152 A red indicator dye in the plate colors the colonies. Count all red colonies regardless of their size or color intensity. 1
3M Petrifilm Aerobic Count Plate 2 3 Count = 0 It is easy to interpret the Petrifilm AC plate. Figure 2 shows a Petrifilm AC plate without colonies. Count = 16 Figure 3 shows a Petrifilm AC plate with a few bacterial colonies. 4 5 Count = 143 The preferable counting range on a Petrifilm AC plate is 25 250 colonies. See figure 4. Estimated Count = 560 When colonies number more than 250, as in figure 5, estimate the count. Determine the average number of colonies in one square (1 cm 2 ) and multiply it by 20 to obtain the total count per plate. The inoculated area on a Petrifilm AC plate is approximately 20 cm 2.
TNTC (Too Numerous to Count) To obtain a more accurate count, dilute the sample further 6 7 Count = TNTC (Estimated count = 10 3 ) Figure 6 shows a Petrifilm AC plate with colonies that are too numerous to count (TNTC). Count = TNTC (Estimated count = 10 5 ) With very high counts, the entire growth area may turn pink, as shown in figure 7. You might observe individual colonies only at the edge of the growth area. Record this as a TNTC result. 8 9 Count = TNTC (Estimated count = 10 3 ) Occasionally, distribution of colonies appears uneven, as shown in figure 8. This is also an indication of a TNTC result. Count = TNTC (Estimated count = 10 7 ) The colonies on the Petrifilm AC plate in figure 9 appear countable at first glance. However, when you look closely at the edge of the growth area, you can see a high concentration of colonies. Record this as a TNTC result.
Gel Liquefication and Food Particles Estimated Count = 160 A few species of bacteria liquify the gel in the Petrifilm AC plate, as shown in figure 10. When this occurs, determine the average count in a few unaffected squares and then multiply it by 20 to obtain the estimated count. Do not count red spots within the liquified area. 10 1 Count = 83 Because colonies on Petrifilm AC plates are red, you can distinguish them from opaque, irregularly shaped food particles (see circles 1 and 2). 2 11
3 Petrifilm Aerobic Count Plates Reminders for Use For detailed CAUTIONS, DISCLAIMER OF WARRANTIES / LIMITED REMEDY, LIMITATION OF 3M LIABILITY, STORAGE AND DISPOSAL information, and INSTRUCTIONS FOR USE see Product s package insert. Storage Petrifilm Petrifilm Petrifilm Store unopened packages at 8 C To seal opened package, fold end 1 2 3 ( 46 F). Use before expiration over and tape shut. date on package. In areas of high humidity where condensate may be an issue, it is best to allow packages to reach room temperature before opening. Keep resealed packages at 25 C ( 77 F) and 50%RH. Do not refrigerate opened packages. Use Petrifilm plates within one month after opening. Sample Preparation Add appropriate quantity of one of the 4 5 6 following sterile diluents: Butterfield's Prepare a 1:10 or greater dilution of food sample. Weigh or pipette food product into an appropriate container such as a stomacher bag, dilution bottle, Whirl-Pak bag, or other sterile container. Inoculation phosphate buffer (IDF phosphate buffer, 0.0425 g/l of KH 2PO 4 adjusted to ph 7.2), 0.1% peptone water, peptone salt diluent (ISO method 6887), buffered peptone water (ISO method 6579), saline solution (0.85-0.90%), bisulfatefree letheen broth, or distilled water. Do not use buffers containing citrate, bisulfite, or thiosulfate; they can inhibit growth. Blend or homogenize sample per current procedure. Adjust ph of the diluted sample between 6.6 and 7.2 : for acid products, use 1N NaOH, for alkaline products, use 1N HCI. Place Petrifilm plate on level With pipette perpendicular to 7 8 9 surface. Lift top film. Petrifilm plate, place 1 ml of sample onto center of bottom film. Release top film; allow it to drop. Do not roll top film down. Continued - over
<70F FLAT side RIDGE side With ridge side down, place Gently apply pressure on spreader to Lift spreader. Wait at least one 10 11 12 spreader on top film over inoculum. distribute inoculum over circular area. minute for gel to form. Do not twist or slide the spreader. Incubation Interpretation 13 14 Incubate plates with clear side up in stacks of no more than 20. It may be necessary to humidify incubator to minimize moisture loss. Petrifilm plates can be counted on a standard colony counter or other magnified light source. Refer to the Interpretation Guide section when reading results. 15 Colonies may be isolated for further identification. Lift top film and pick the colony from the gel. Incubation time and temperature varies by method. Most common approved methods: AOAC Official Method 986.33 & 989.10 (milk and dairy products) Incubate 48h ± 3h at 32 C ± 1 C AOAC Official Method 990.12 Incubate 48h ± 3h at 35 C ± 1 C AFNOR Validated Method 3M 01/1-09/89 Incubate 72h ± 3h at 30 C ± 1 C Additional Comments Questions? U.S., call 1-800-328-6553, Canada, call 1-800-563-2921 for technical service. To order Petrifilm plates in the U.S., call 1-800-328-1671. Latin America / Africa regions, call 5255-5270-0454 3 Microbiology Products 3M Center Bldg. 275-5W-05 St. Paul, MN 55144-1000 USA 1 800 328-6553 www.3m.com/microbiology Email: microbiology@3m.com 3M Canada Post Office Box 5757 London, Ontario N6A 4T1 Canada 1 800 563-2921 Asia Pacific region, call 65-64548611 3M Europe Laboratoires 3M Santé Boulevard de l Oise F-95029 Cergy-Pontoise Cedex France 33 1 30 31 8571 3M and Petrifilm are trademarks of 3M Company. Whirl-Pak is a registered trademark of Nasco. 40% Pre-consumer waste paper 10% Post-consumer waste paper Printed in U.S.A. 2005 70-2008-4572-8 (15.35)ii