3M Food Safety
3M™ Petrifilm™ Aerobic Count Plate
Aerobic
Count
Interpretation Guide
The 3M™ 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. 3M Petrifilm AC Plates are
used for the enumeration of aerobic bacteria.
It is easy to interpret the 3M Petrifilm AC Plate. A red indicator
dye in the plate colors the colonies. Count all red colonies
Aerobic Bacteria Count: 152 regardless of their size or color intensity.
Interpretation Guide: 3M™ Petrifilm™ Aerobic Count Plate
3M™ Petrifilm™ Aerobic Count (AC) Plates
Figure 1 Figure 2
Count: 0 Count: 16
3M Petrifilm AC Plate without colonies. 3M Petrifilm AC Plate with a few bacterial colonies.
Figure 3 Figure 4
Count: 143 Count: ~560
The preferable counting range on a 3M Petrifilm AC Plate is When colonies number more than 250, estimate the count.
25–250 colonies. Determine the average number of colonies in one square (1cm2) and
multiply it by 20 to obtain the total count per plate. The inoculated
area on a 3M Petrifilm AC plate is approximately 20cm2.
User’s Responsibilities: 3M Petrifilm Plate performance has not been evaluated with all combinations of microbial flora, incubation conditions and food matrices. It is
the user’s responsibility to determine that any test methods and results meet the user’s requirements. Should re-printing of this Interpretation Guide be necessary, user’s
print settings may impact picture and color quality.
2
TNTC (Too Numerous to Count) To obtain a more accurate count, dilute the sample further.
Figure 5 Figure 6
Count: TNTC (Estimated count = 103) Count: TNTC (Estimated count = 105)
3M Petrifilm AC Plate with colonies that are TNTC. With very high counts, the entire growth area may turn pink. You
might observe individual colonies only at the edge of the growth
area. Record this as a TNTC result.
Figure 7 Figure 8
Count: TNTC (Estimated count = 103) Count: TNTC (Estimated count = 107)
Occasionally, distribution of colonies appears uneven. This is also The colonies on the 3M Petrifilm AC Plate may appear countable
an indication of a TNTC result. 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.
User’s Responsibilities: 3M Petrifilm Plate performance has not been evaluated with all combinations of microbial flora, incubation conditions and food matrices. It is the
user’s responsibility to determine that any test methods and results meet the user’s requirements. Should re-printing of this Interpretation Guide be necessary, user’s print
settings may impact picture and color quality.
3
Gel Liquefication and Food Particles
Figure 9 Figure 10
Count: ~160 Count: 83
A few species of bacteria liquify the gel in the 3M Petrifilm AC Because colonies on 3M Petrifilm AC Plate are red, you can
Plate. When this occurs, determine the average count in a few distinguish them from opaque, irregularly shaped food particles.
unaffected squares and then multiply it by 20 to obtain the See Circles 1 and 2.
estimated count. Do not count red spots within the liquified area.
User’s Responsibilities: 3M Petrifilm Plate performance has not been evaluated with all combinations of microbial flora, incubation conditions and food matrices. It is
the user’s responsibility to determine that any test methods and results meet the user’s requirements. Should re-printing of this Interpretation Guide be necessary, user’s
print settings may impact picture and color quality.
4
Reminders for Use: 3M™ Petrifilm™ Aerobic Count (AC) Plate
Storage
Petrifilm Aerobic
Petrifilm Aerobic
™
≤8°C Petrifilm Aerobic
™
1 Store unopened pouches of plates at <8°C
(<46°F). Use before expiration date on package.
2 To seal opened pouch, fold end over and
tape shut.
3 To prevent exposure to moisture, do not
refrigerate opened pouches. Store resealed
In areas of high humidity where condensate may pouches in a cool, dry place. Use plates within
be an issue, it is best to allow pouches to reach one month after opening. Avoid exposure of plates
room temperature before opening. to temperatures >25°C (>77°F) and/or relative
humidity >50%.
Sample Preparation
4 Prepare a dilution of food product.* Weigh or
pipette food product into an appropriate
5 Add appropriate quantity of one of the following
sterile diluents: Butterfield’s phosphate buffer
6 Blend or homogenize sample per current
procedure. For optimal growth and recovery of
container such as a stomacher bag, dilution bottle, (IDF phosphate buffer, KH2PO4 @ 0.0425 g/L, microorganisms, adjust the pH of the sample
Whirl-Pak® bag, or other sterile container. adjust to pH 7.2), 0.1% peptone water, peptone suspension to 6.6–7.2:
*See Petrifilm Use with Dairy and Juice Products sheet
salt diluent (ISO method 6887-1), buffered peptone • For acidic products, adjust the pH with 1N NaOH
for recommended dilutions. water (ISO 6887-1), saline solution (0.85–0.90%), • For alkaline products, adjust the pH with 1N HCl
bisulfite-free letheen broth or distilled water. Do
not use buffers containing citrate, bisulfite or
thiosulfate; they can inhibit growth.
Inoculation
7 Place 3M Petrifilm Aerobic Count Plate on level
surface. Lift top film.
8 With 3M™ Electronic Pipettor or equivalent held
perpendicular to plate, place 1mL of sample or
9 Roll top film down onto sample gently to prevent
pushing sample off film and to avoid entrapping
diluted sample onto center of bottom film. air bubbles. Do not let top film drop.
10 With ridge side down, place 3M™ Petrifilm™
Spreader on top film over inoculum.
11 Gently apply pressure on 3M Petrifilm Spreader
to distribute inoculum over circular area before
12 Lift 3M Petrifilm Spreader. Wait a minimum of
1 minute for gel to solidify.
gel is formed. Do not twist or slide the spreader.
5
Reminders for Use: 3M™ Petrifilm™ Aerobic Count (AC) Plate cont.
Incubation Interpretation
13 Incubate plates with clear side up in stacks of up
to 20 at time and temperature listed below. It may
14 3M Petrifilm Aerobic Count Plates can be counted
on a standard colony counter or other illuminated
15 Colonies may be isolated for further identification.
Lift top film and pick the colony from the gel.
be necessary to humidify incubator to minimize magnifier. Refer to the Interpretation Guide
moisture loss. section when reading results.
Incubation Time and Temperature Vary by Method
Most common approved methods:
AOAC® Official Method 986.33 and 989.10 (milk and dairy products)
• Incubate 48±3 hours at 32°±1°C
AOAC® Official Method 990.12
• Incubate 48±3 hours at 35°±1°C
AFNOR Validated Method 3M 01/1-09/89
• Incubate 72±3 hours at 30°±1°C
For detailed CAUTIONS, DISCLAIMER OF WARRANTIES/LIMITED REMEDY and LIMITATION OF 3M LIABILITY,
STORAGE AND DISPOSAL information and INSTRUCTIONS FOR USE, see Product’s package insert.
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