Interpretation

Guide
The 3M™ Petrifilm™ Coliform Count Plate
is a sample-ready-culture medium system
which contains modified Violet Red Bile
nutrients, a cold-water-soluble gelling agent
and a tetrazolium indicator that facilitates
colony enumeration.

CC
Coliform Count Plate
Food and Beverage Applications
Coliform Definitions by Method
The United States Food and Drug Administration (FDA) Bacteriological
Analytical Manual (BAM) defines coliforms as Gram negative rods, which
produce acid and gas from lactose fermentation. Coliform colonies growing
on the 3M™ Petrifilm™ Coliform Count Plate produce acid, which causes
the pH indicator to deepen the gel color, and gas trapped around red
colonies. In this interpretation guide, the number of coliforms per the FDA
BAM definition is the number of red colonies with gas.
ISO defines coliforms by their ability to grow in method-specific, selective
media. ISO method 4832 enumerates typical coliform colonies on Violet
Red Bile Lactose (VRBL) agar, with confirmation of atypical colonies. On
the 3M Petrifilm Coliform Count Plate, these coliforms are indicated by red
colonies with or without gas production. ISO method 4831, enumerating
coliforms by the most probable number (MPN) method, defines coliforms
by their ability to grow and produce gas in the conditions described in the
standard. On the 3M Petrifilm Coliform Count Plate, these coliforms are
indicated by red colonies with gas.
It is also possible to enumerate thermotolerant coliforms on the 3M
Petrifilm Coliform Count Plate. Typically thermotolerant coliforms can
be selected with an elevated incubation temperature. One example of a
method for enumeration of thermotolerant coliforms is described in NF Please refer to the
V08 060. Reading the total of red colonies on a 3M Petrifilm Coliform product instructions for
Count Plate incubated at 44°C ± 1°C for 24h ± 2h yields results equivalent additional information.
to enumeration with NF V08 060.

Figure 1 Figure 2

Total colonies with gas = 69 No growth = 0

Total colonies = 94 Notice the changes in gel color in Figures 2–5. As the coliform
The definition of coliforms may vary by country. count increases, the gel color deepens.
Please refer to section above and product instructions Background bubbles are a characteristic of the gel and are not
for definitions. a result of coliform growth.

Coliform Count Plate 2

 1
Figure 3 Figure 4

Total colonies with gas = 79 Estimated total coliform count = 220

Total colonies = 109 The circular growth area is approximately 20cm2. Estimates can be
The recommended counting limit on 3M Petrifilm Coliform made on plates containing greater than 150 colonies by counting
Count Plates is less than 150. the number of colonies in one or more representative squares and
Do not count colonies that appear on the foam barrier because determining the average number per square. Multiply the average
they are removed from the selective influence of the medium number by 20 to determine the estimated count per plate.
(see Circle 1). For a more accurate count, further dilution of sample may
be necessary.

Figure 5 Figure 6

Total coliform count = TNTC Total count = TNTC

3M Petrifilm Coliform Count Plates with colonies that are too When high numbers of non-coliform organisms such as
numerous to count (TNTC) have one or more of the following Pseudomonas are present on 3M Petrifilm Coliform Count
characteristics: many small colonies, many gas bubbles, and a Plates, the gel may turn yellow.
deepening of the gel color.
For a more accurate count, further dilution of sample may
For a more accurate count, further dilution of sample may be necessary.
be necessary.

3
 1

3
2

Figure 7 Figure 8

Total colonies with gas = 2 Total colonies with gas = 8

Total colonies = 2 Total colonies = 15
Food particles are irregularly shaped and are not associated Bubble patterns may vary. Gas may disrupt the colony so that
with gas bubbles. the colony “outlines” the bubble (see Circles 1 and 2). Artifact
bubbles may result from improper inoculation or from trapped
air within the sample. They are irregularly shaped and are not
associated with a colony (see Circle 3).

Bottled Water Applications

Coliform colonies are indicated by red colonies associated with gas for bottled water samples plated to 3M
Petrifilm Coliform Count Plates.

Figure 1 Figure 2

Coliform count: 0 Coliform count: 0

Gas bubbles surrounding filter do not indicate microbial growth. Red colonies without closely associated gas bubbles may be
See circle for example. coliforms and should be picked and tested with appropriate
confirmation methods.

4
 Figure 3 Figure 4

Coliform count: 3 Coliform count: 10

Coliforms produce acid (faint pink halo associated with colonies)
and are associated with gas bubbles.

C
A

Figure 5

Coliform count: 30
Gas bubbles may influence colony morphologies. The colony in
Circle A is distorted by the gas bubble. In Circle B, a faint colony
is underneath the gas bubble. Note large artifact gas bubble in the
center of the plate (Circle C).

5
 Reminders for Use:
Food and Beverage Applications
Storage Inoculation

Petrifilm Coliform
™

≤8°C

Store unopened pouches of plates at To seal opened pouch, fold end over and Place 3M Petrifilm Coliform Count Plate
1 <8°C (<46°F). Use before expiration date 2 apply adhesive tape. 3 on level surface. Lift top film. With a
on package. In areas of high humidity pipette perpendicular to plate, place 1mL
where condensate may be an issue, it of sample or diluted sample onto center
is best to allow pouches to reach room of bottom film.
temperature before opening.

Roll top film down onto sample gently With flat side down, place 3M™ Petrifilm™ Gently apply pressure on 3M Petrifilm
4 to prevent pushing sample off film and to 5 Spreader on top film over inoculum. 6 Spreader to distribute inoculum over
avoid entrapping air bubbles. Do not let circular area before gel is formed.
top film drop. Do not twist or slide the spreader. Lift
3M Petrifilm Spreader. Wait a minimum
of 1 minute for gel to solidify.

Incubation Interpretation Use Appropriate

Sterile Diluents
Butterfield's phosphate buffered dilution
water, 0.1% peptone water, peptone salt
diluents, buffered peptone water, saline
solution (0.85–0.90%), bisulfite-free
letheen broth or distilled water.

For optimal growth and recovery of the

microorganisms, adjust the pH of the
sample suspension to 6.6-7.2.

Do not use diluents containing citrate,

Incubate plates with clear side up in 3M Petrifilm Coliform Count Plates can
7 stacks of up to 20. It may be necessary 8 be counted using the 3M™ Petrifilm Plate
bisulfite or thiosulfate with the 3M
to humidify incubator to minimize Reader, on a standard colony counter or Petrifilm Coliform Count Plates; they
Printed Colors – Front:
moisture loss. See product instructions other illuminated magnifier. Colonies may can inhibit growth.
Requester: forBarker
Susan third party validated methods. be isolated for further identification. Lift
Creator:
File Name:
deZinnia_21849
Petrifilm_21849-07_FS.ai
top film and pick the colony from the gel. If citrate buffer is indicated in the
Scale:
Structure #: Illustration 1 Inch standard procedure, substitute with one
Date: 04/17/17
of the buffers listed above, warmed to
This artwork has been created as requested by 3M. 3M is responsible for the artwork
AS APPROVED and assumes full responsibility for its correctness. 40–45°C.

Coliform Count Plate 6

Reminders for Use:
Bottled Water Applications
Storage Hydration Procedure

≤8°C

Store unopened packages at ≤8°C To seal opened package, fold end over and Place 3M Petrifilm Plate on a flat, level
1 (≤46°F). Use before expiration date on 2 apply adhesive tape. Do not refrigerate 3 surface. With the pipette perpendicular
package. Just prior to use, allow unopened opened packages. Use 3M Petrifilm to the 3M Petrifilm Plate, place hydration
pouches to come to room temperature Coliform Count Plates within one month diluent onto the center of the bottom film.
before opening. after opening. Hydration diluents include distilled water,
deionized (DI) water and reverse osmosis
(RO) water.

Carefully roll top film down to avoid With flat side down, place 3M™ Petrifilm™ Gently apply pressure on spreader to
4 entrapping air bubbles. Do not let top 5 Spreader on top film over hydration 6 distribute inoculum or hydration diluent
film drop. diluent. over circular area before gel is formed. Do
not twist or slide spreader. Lift spreader.
• Allow the hydrated plates to remain
closed for a minimum of one hour
before use.
• Any additional hydrated 3M Petrifilm
Coliform Count Plates may be
stored in a sealed pouch or plastic
bag. Protect plates from light and
refrigerate at 2–8°C (36–46°F) for up
to 7 days.

Following standard procedures for water Carefully lift top film.

7 analysis, membrane filter water sample 8
using a 47 mm, 0.45 micron pore size
Mixed Cellulose Ester (MCE) filter.

Place filter in the center of the well. Roll top Lightly apply pressure to ensure uniform
9 film down to minimize air bubbles or gaps
10 contact of the filter with the gel and to
between the filter and the 3M Petrifilm eliminate any air bubbles.
Coliform Count Plate.

7
Incubation Interpretation

Incubate 3M Petrifilm Coliform Count 3M Petrifilm Coliform Count Plates can Colonies may be isolated for further
11 Plates in a horizontal position, clear side 12 be counted on a standard colony counter
13 identification. Lift top film and pick the
up, in stacks on no more than 20 plates or other illuminated magnifier. colony from the gel. Red colonies without
at 35°± 1°C for 24 ± 2 hours or 36°± closely associated gas bubbles may be
1°C for 24 ± 2 hours. Please refer to the coliforms and should be picked and tested
the product instructions for third party with appropriate confirmation methods.
validated methods.

Bubbles
The illustrations below show examples of various bubble patterns
associated with gas producing colonies.
All should be enumerated.

3M Food Safety offers a full line of

products to accomplish a variety of
your microbial testing needs. For
more product information, visit us at
3M.com/foodsafety/Petrifilm or call
1-800-328-6553.

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.

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3M Center, Building 275-5W-05 Post Office Box 5757 and LIMITATION OF 3M LIABILITY, STORAGE AND DISPOSAL information
St. Paul, MN 55144-1000 USA London, Ontario N6A 4T1 and INSTRUCTIONS FOR USE, see Product’s package insert.
Canada 3M and Petrifilm are trademarks of 3M. Used under license in Canada.
1-800-328-6553 Please recycle. Printed in USA. © 3M 2020. All rights reserved.
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