3MTM

3MTM

3MTM

3MTM

3MTM

3MTM

3MTM

FDA BAM 3MTM

3MTM

3MTM Food Safety Department ()

3MTM

3MTM

3MTM FAQ

3MTM


.
Biological film

dry rehydratable film media. 3MTM Petrifilm

. 3MTM Petrifilm

.
3MTM

3MTM (Petrifilm) cold water soluble gel

film (dry
rehydratable film media) , , .

3M , 2 .
, + , , + , ,
.
3MTM

7.
8.

2) ()

1) (1) 1 1
( 53) 351 24~48
.

1
1
1 (1g )
. "" .
3 4 5 2 0 .

5)
1) (1) 1 1
( 54) , 351 242
,
.

6)
1) (1) 1 1
( 55) , 351 24~48

.

3.
III.
9.
.
1)
)
. 2. . .
) : Butterfield's phosphate buffered dilution water(BPD)
. 1. . .
2)
. (2) 1 1
351 24~48
.
3)
. 9. (1) () )
.

.
. (2) 1 1
, 351 242
, .

.
. (2) 1 1
, 351 24~48
.
3MTM

Standard Nutrient TTC* (

(Petrifilm - )

Aerobic Count

Plates)

Violet Red Bile Lactose TTC* (

(Petrifilm - )

Coliform Count Gas CO2

Plates)

Violet Red Bile Lactose TTC (

(Petrifilm E. - )

coli/Coliform BCIG* Glucuronidase

Count Plates)

Gas CO2

Potato Dextrose with BCIP - Phosphatase

(Petrifilm Yeast Antibiotics* ()

and Mold Count

Plates)

Modified Baird Parker TTC (

(Petrifilm Staph - )

Express Count Toluidine blue-O Dnase

Plates)

PALCAM,

(Petrifilm Modified Oxford(MOX),

Environmental IDF pre-enrichment broth, LRB

Listeria Plates)

Violet Red Bile Lactose TTC* (

(Petrifilm Rapid - )

Coliform Count Methyl Red

Plates) Gas CO2

Violet Red Bile Glucose TTC* (

(Petrifilm - )

Enterobacteriac Phenol Red

eae Count Gas CO2

Plates)

Modified Violet Red Bile BCIG* Glucuronidase

Lactose

Violet Red Bile Lactose TTC* (

- )

(High Sensitivity Gas CO2

Coliform Count

Plates)

*
TTC: Tri-phenyl-tetrazolium chloride
BCIG: 5-bromo-4-chloro-3-indoxyl--D-glucuronide
BCIP: 5-bromo-4-chloro-3-indoxyl phosphate
PYM : chloromaphenicol chlorotetracycline
3MTM

() ()
35 24~48 AOAC 482
35 24
35 24~48
25 72~120
35 24 Dnase 3
Dnase 35 1~3
35 282
35 8~24
35 24
35 24



.

.
(180) 1 90
60.7% .
.
(
, ) (53) ( 4 )
.

.
.
, , 1
.

2 .
2
.

(homogenizer stomacher)
.
,
.

.
.
.
(1025
) 9 .
(1025g)
.
(100250) .
 ( 100cm2) (15)
(10100)
.
(10g)
9 .
40 15 10
100 .
(1025g) 9
.
10 , 100 , 1,000
.
510cm 70% ,
, 2 . Tween
80 .
40
, 70% .
3MTM

: .

1. 2. (Phosphate 3.
buffer, 0.1% peptone water, .
distilled water, phosphate
buffered saline, and
Butterfields buffer)
( ).

4. 5. 20 6. Technical guide book

(1ml (PAC, PCC, PEC, PEB, . .
STX, PYM) or 3ml (RCC) or 5ml
(PEL))
3MTM

. .
3MTM .

= ( X )

Ex) 7.5 ml 1 : 10

( 7.5 ml X 10 ) - 7.5 ml = 67.5 ml

( ) () ( ) ( )


colony counter PPR .

5 24 . ( 1/2
) 1 30300
. 300 300
1cm2 20 .
30 .

.
1

1 (1g )
. ""
. 3 4 5 2 0 .

10-1 10-2 10-3 CFU/g

TNTC 160 14 18,000 30~300

TNTC 200 21
TNTC 222 36 29,000 2 30~300
TNTC 244 32

24 4 0 400 30
21 0 0

TNTC TNTC 500 47,000 300

TNTC TNTC 440

0 0 0 N.D.
0 0 0

TNTC : Too Numerous To Count ( )

N.D. : Not Detected
3MTM

Aerobic Count Air sampling Direct 1ml
Coliform Count Contact Method plate . 1
E. coli/Coliform Count plate .
Rapid Coliform Countt
Enterobacteriaceae Count
Staph Express Count Air sampling Direct 1ml
Contact Method plate . 3
plate .
Yeast and Mold Count Air sampling Method 1ml
plate . 1
plate .
Yeast and Mold Count Direct Contact Method Yeast and Mold plate 1ml letheen
broth .
plate 30-37 24
4
.
Air sampling Method

1. clip 2. , clip
. .

3. clip 4. 3MTM
. 15 .

.
Direct Contact Method

1. 2a.
. .
.

2b. 3. 3MTM Petrifilm

. .
.
.


3MTM

Petrifilm Aerobic Count

plates

, , , , , Petrifilm Yeast and Mold

( ) High sugar, , , Count Plates

High salt

, , , , Petrifilm Coliform Count

, Plates

, , , Petrifilm Rapid Coliform

Count Plates

, , Petrifilm High-Sensitivity
Coliform Count Plates

, , , , Petrifilm E. coli/Coliform
Count Plates

, , Petrifilm Enterobacteriaceae
Count Plates

, , , , Sraph Express Count Plates

, ,

, , , Petrifilm Environmental
Listeria Count Plates
 FDA (Food & Drug Administration) BAM (Bacteriological
Analytical Manual) 3MTM

Nutrient 48 at 35 2
Petrifilm 48 at 35 2

PDA 5 at 25 5
5
Petrifilm 5 at 25 - 5

VRBA 24 at 35 Illumination
BGLB 24~48 at 35
Gram
3

LST 3 each dil 24~48 at 35
BGLB broth 48 at 35 (MPN/g)
Gram
4

Petrifilm 24 at 35

24

LST/BGLB 48 EC broth streaking
EC Broth 24~48 at 45.5
MPN
L-EMB 24 at 35

PCA slalnt 24 at 35 Bio(IMVIc),
4~6

Petrifilm 48 at 35
2

BP 48 at 35 3 ,
,
BHI Broth 24 at 35 Purify & Enumerate
Coagulase 6 at 35
4

Petrifilm 26 at 35

1


(10-1) 4 at 30- acriflavin, nalidixic
acid, cycloheximide 44 at 30
PALCAM OXA 24~48 at 35 Black Halo
TSAYE (5) 24~48 at 30
BioChem& Rapid Kit , Henry, NO2 illumination,
motility, CHO ferment, Elisa, Serology
6~7

Petrifilm 31 at 35
31

VRBG 24 at 37 Brack red SMEDP
BGGB (MPN) 48 at 37 ISO 7402
Petrifilm 24 at 35

BAM 3MTM
FDA BAM
Nutrient Agar
Potato Dextrose Agar (PDA)
VRB Agar
Baird Parker Agar (BP Agar) & Coagulase
MPN-EMB-PCA Slant
ISO 11290-2
3MTM

00 11 22 33

()
()
3M Food Safety Department()

1. 3MTM
3MTM dry rehydratable film media
.
.

= =
(Efficiency) (Productivity) (Profitability)

3M 1
. 1500 1
1,405 1 1,353
52 .
( : )
3M
(/)
1 () 15,000,000 15,000,000
(/) 12,207 12,207
(/) 14.8 36.7
() 825 333
(/)
( ) 150 150
110 0
160 800
420 950

(/) 160 70
( )

()/ 1,405 1,353

(/)= 52
1 5 = 2,609,138
, 1,340 1 , 1,317
 . ,
.
, 3MTM 4 .

(Increased Efficiency) (Increased Productivity)

(Decreased Labor hour) (Decreased Cost)

3MTM .
variability .
, water bath, .
.
.
.
, , AOAC .
.

2. 3MTM Petrifilm Plate Reader (PPR)

3MTM Petrifilm Plate Reader( PPR) 3MTM . PPR
3MTM ,
.
PPR .

, , /
.
.
IAFP 3 20 manual
.
1 4 .
,
.
.
ID
..
, .
3MTMPPR Data time saving, transcription error, data
, .
Microsoft Excel sheet uneditable log file transcription error
. Transcription error
.
Unchangeable log file , FDA 12CFRPart11
.
Log file text LIMS (Lab information
management system) .
Mapping
.

.

PPR .
MicrosoftWindows 2000/XP
Microsoft Excel 2000 or higher (to use spreadsheet option)
USB2 Port
256MB RAM
CD-Rom Drive
1.5GHz CPU
5GB disk space needed (to allow for saving data and images)
3M PPR 3MTM
powerful , process .

9









3M

5
1)
2)
3)
4)
5) PPR

3M PPR , , PPR
LIMS(Laboratory Information Management
System) software. PPR 3M
, PPR
.
3MTM PPR

() 3M
()
110 0
50 50
65 24
5 1
/ 72 9
25 0
8 5

() 335 89
() 5.6 1.5
* 1 100
1 () 1,049.6
1 () 131.2
* 1 8

, 1 100 3M 3M PPR 1
131 . 5 250
3M 131
.
 PPR .
160 PPR .
4.2 PPR , 1,240
. , ,
, ,
.

3M Petrifilm Plate Reader Benefit Analysis Input Worksheet/Overall Results

160

3.2

8,000

- % 72.00 %

1.333333333

8,000

- % 72.00 %

$13,760.00

3M Petrifilm Plate Reader 4,400,000

- 12,974,763

- 3M Petrifilm Plate Reader 508,814

3M Petrifilm Plate Reader = 906

3M Petrifilm Plate Reader = 12,465,948

PPR () 4.2
3MTM

(1)
8 .
.
(2)
.
.
, .

. 25
50% .
.
.
.

1.
8 .

(PAC) 18
(PCC) 18
/ (PEC) 18
(PYM) 18
(RCC) 12
(PEB) 18
(HSCC) 18
(STX) 18
(PEL) 18
Note: Lot .
3MTM

.
1) .
2) .
3) .

Option 1: Autoclave or bench-top sterilizer

Autoclaving of inoculated Petrifilm Plates prior to disposal is recommended. A typical
autoclave protocol of 121/250 and 15 pounds per square inch (psi) is appropriate for
decontaminating inoculated Petrifilm Plates. Most lab supply houses sell portable
sterilizers and autoclaves that range from $500 to $50,000.
Option 2: Contract waste disposal service
Professional disposal services may be contracted for removal of biological waste, such as
inoculated Petrifilm Plates.
Option 3: Pressure cooker
Five- to 12-quart pressure cookers can be purchased for less than $100 from most
discount or farm stores. A lab hotplate may be required for heating the pressure cooker.
Follow the operators instructions when using the pressure cooker.
1. Place Petrifilm Environmental Listeria Plates into a suitable heat resistant container
(e.g., metal or polypropylene)
2. Add water to the pressure cooker to produce steam
3. Process at a minimum of 10 psi (115.6/240)*
4. Process for a minimum of 30 minutes

o It is important to begin the 30-minute timing AFTER the unit reaches a

minimum of 10 psi (which is when pressure controller jiggles)
o The time required to reach 10 psi can vary depending on the amount of material inthe
pressure cooker
*CRC Handbook of Chemistry and Physics, 54th Edition, E-18 Steam Tables
Handbook of Chemical Engineering Calculations 2nd Edition, 1994. Ed. N.P. Chopey, McGraw Hill
Inc.
3MTM FAQ

Q) 3MTM ?
A) 3MTM , ,
gelling agent .
, 3MTM
.
.

Q) 3MTM ?
A) 3MTM .
. ,
3MTM ,
. ,
. 3MTM
.

Q) 3MTM ?
A) 3MTM AOAC . AOAC
, MPN() FDA(Food Drug
Administration) BAM(Bacteriological Analytical Manual)
.

Q) 3MTM ?
A) 3MTM () (
) , AOAC, AFNOR
. 75% 3MTM
.

Q) 3MTM ?
A) 3MTM , , .
3M ,
. , QC
.
Q) , O157:H7, , , 3MTM
?
A) 3MTM , O157:H7, ,
, . 3M

. , 3M .

Q) 3MTM , , O157:H7 ?
A) 3MTM O157:H7, ,
. 3MTM
,
. , 3MTM
.

Q) 3MTM . ?
A) 3MTM 100 .
3M Korea Technical Bulletin TB-001 Shipping and Receiving
.

Q) ?
A) , .
(, ) . , 1:10
.
. , .
: Butterfields phosphate water, buffered peptone water,
quarter-strength Gingers solution, saline solution (0.85~0.90%), bisulfate-free Letheen broth.

Q) pH ?
A) pH , /, , ,
pH .
. pH
. .
. 3M
pH 7 . , ,
6.5~7.5 , 6~8 . pH
7.0~7.2 pH .
pH 3M
pH .
 pH , pH
.

Q) pH ?
A) pH pH NaOH pH HCl .

Q) pH ?
A) pH . pH
pH strip , pH meter
. pH , pH .
3MTM

Petrifilm Aerobic Count Plate (PAC)

.

.

Pancreatic Digest of Casein 3.4g

Yeast Extract 2.4g
Sodium Pyruvate 6.8g
Dextrose 0.6g
Potassium Phosphate(dibasic) 1.3g
Potassium Phosphate(monobasic) 0.4g

Guar Gum(Cold water soluble gelling agent) 91.4g

2,3,5-Triphenyltetrazolium Chloride 0.0205g
(Tetrazolium indicator dye for red color reaction)

1) 20-25
251 723
.

2)
pasteurization . 351 483
.
 Genera/group Associated Foods
Actinomycetes Milk
Alcaligenes Milk
Arthrobacter Milk
Bacillus Milk/eggs
Clostridium Milk
Coryneform bacteria Eggs
Lactobacillus Milk/meats/juices
Microbacterium Milk
Micrococcus Milk
Psuedomonas Eggs
Streptococcus/Enterococcus Milk/meats/eggs

* From Compendium of Methods for the Microbial Examination of Foods , by the American
Public Health Association

3) , .
MRS broth . 35-37 723 .

Liquifier
1) (Liquifiers)
, Agar plates . ,
.
AOAC .
) 16
= 16 CFU/square/g 20 squares = 320 CFU/g
2)
- .
- 24 48 2 .
- 5-10 plate
- 30 ~ 32
-
Petrifilm Coliform Count plates (PCC)

Gram , (lactose)
. 1g
108-11
CFU (colony forming unit) , ,
. , . , , ,
. E. coli, Klebsiella pneumoniae(
), Enterobacter cloacae & aerogenes( ), Erwinia carotovora &
Citrobacter freundii(, , ) 6,600
.

Yeast Extract 9.6g

Pancreatic Digest of Gelatin 20.9g
Bile Salt #3 1.6g
Peptic Digest of Animal Tissue 1.6g
Lactose 21.4g
Sodium Chloride 5.3g
Crystal Violet 2
Neutral Red 0.1g
Guar gum(Cold water soluble gelling agent) 65.7g
2,3,5-Triphenyltetrazolium Chloride 0.11g

1)
43~45
.
44-44.5 .
Petrifilm Rapid Coliform Count plates (RCC)

.

. .

3M .
Petrifilm E. coli/Coliform Count plates (PEC)

/ .
.
. ,
.
(undercook
etc.) (post-contamination) . EPEC, ETEC, EIEC,
EHEC . 3M -glucuronidase indicator -glucuronidase
. chromogenic media E. coli
O157 -glucuronidase .

Yeast Extract 9.6g

Pancreatic Digest of Gelatin 20.9g
Bile Salt #3 1.6g
Peptic Digest of Animal Tissue 1.6g
Lactose 21.4g
Sodium Chloride 5.3g
Crystal Violet 2
Neutral Red 0.1g
Guar gum 65.4g
5-Bromo-4-Chloro-3-Indoxyl-beta-D-Glucuronic Acid, 0.2g
Cyclohexylammonium Salt
2,3,5-Triphenyltetrazolium Chloride 0.11g
Petrifilm Staph Express Count plates and Disk

.
19
.
.

Modified Baird Parker agar

3M

Dried potato flakes 1:10 1:100 .
.

Whey powder 1:10 S. aureus Staph Express disk

1:100
. .
Soy flour 1:10 S. aureus Staph Express disk
1:100
. .

Beef gravy mix 1:10 Staph Express disk

1:100
.
.

Dried orange or red cheese 1:10 1:100 .

powder

.

Dried dark chocolate mix 1:10 Staph Express disk

1:100
.
.

Raw milk Raw milk 1:10
phosphatase .
pink
,
.
.
Raw milk cheese Raw milk 1:10
phosphatase . , Staph
pink , Express disk
.
. .
Raw shrimp Disk Pink zone S. aureus ,
. Coagulase test .
Meat trimmings . Red-violet colony
Staph
Express disk
.
Refrigerated dough . Red-violet colony
Staph
Express disk
.
Raw products/In-process . Red-violet colony
foods Staph
Express disk
.
 (STX) .

Q) Staph Express plate(STX) ?

A) .
. S. aureus
. S. aureus (+),
.

Q) Disk ?
A) . S.
aureus .

Q) Disk ?
A) S. aureus Disk DNA .
DNA pink zone .

Q) Disk ?
A) Red-violet .
disk .

Q) STX ?
A) .
.

Q) STX red-violet ?
A) S. aureus.

Q) STX blue-green ?
A) Blue-green Enterococcus or Bacillus .

Q) STX black ?
A) Black staphylococcus , stress S. aureus .
.
Q) Environmental testing STX ?
A) STX , swab test .

Q) S. aureus pink-zone ?
A) Pink-zone S.aureus . Staphylococcus hyicus, Staphylococcus intermedius
pink-zone , enterotoxin
.

Q) surface sampling STX ?

A) surface sampling STX . sampling 3
. 7 .

Q) ?
A) 1 .

Q) disk ?
A) 5 disk .
.

Q)
?
A) shelf-life . plates disk
disk . (2 )
.

Q) disk ?
A) pouch disk disk
. disk .

Q) STX enterotoxin ?
A) . . Enterotoxin
, S. aureus protein .
Petrifilm Enterobacteriaceae Count plates (PEB)

. ,
, , , , , , .
, .
. , .


.

Violet red bile/glucose nutrients

Petrifilm Environmental Listeria plates (PEL)

Listeria . Listeria monocytogenes ,

. , .
Listeria Listeria monocytogenes, Lsiteria innocua, Listeria welshimeri and Listeria
grayilmurrayi 8 , (listeriosis) Listeria
monocytogenes 1 . , , ,
30% , 70% .

Listeria .

PALCAM
Modified Oxford (MOX) medium
IDF pre-enrichment broth
Listeria repair broth (LRB)
 Listeria

3M Petrifilm, Environmental Listeria Plate Listeria monocytogenes Listeria

. 2,600 . Listeria
L. monocytogenes, L. innocua, L. welshimeri 98% , 2%
L. grayi, L. ivanovii, L. seeligeri .

Listeria species

(Food, Food environments, Feed) (Percentage of Listeria-positive sample)

Raw refrigerated ground beef, pork 180 L. innocua 45.4%

sausage, ground turkey, chicken L. monocytogenes 40.2%

L. welshimeri 14.6%

Food processing environmental 552 L. monocytogenes 89.4%

L. innocua 8.8%

L. seeligeri 0.9%

L. ivanovii 0.9%

Seafood 128 L. innocua & L. welshimeri 74.1%

L. monocytogenes 25.9%

Smoked fish 258 L. innocua 42%

L. welshimeri 30%

L. monocytogenes 28%

Crawfish plants-whole, raw crawfish 78 L. innocua 66.6%

L. monocytogenes 14.8%

L. welshimeri 14.8%

L. grayi 3.7%

Crawfish plants-environment 407 L. innocua 60.8-74.1%

L. monocytogenes 13.0-18.9%

L. welshimeri 3.4-13.0%

L. grayi 1.7-4.3%

L. welshimeri/innocua 1.7-8.7%

Animal feed-corn, hay and grass silage 210 L. innocua 83.7%

L. monocytogenes 13.5%

L. welshimeri 2.7%

Dairy plants-environment and 378 L. innocua 60%

equipment L. monocytogenes 35%

 L. welshimeri 5%

Raw vegetables, beef, chicken, pork, 410 L. innocua 71.7%

frozen foods L. monocytogenes 17.4%

L. seeligeri 6.5%

L. welshimeri 4.4%

1. Elliot T. Ryser, Samuel M. Arimi, M. Marie-Claire Bunduki & Catherine W. Donnelly, 1996.
Recovery of different Listeria ribotypes from naturally contaminated, raw refrigerated
meat and poultry products with two primary enrichment media. Applied &
Environmental Microbiology, 62 (5): 1781-1787.
2. Joseph A. Odumeru, Jennifer Belvedere, Cara van Donkersgoed, 2004. Evaluation of the
MicroFoss system for the detection of Listeria species in environmental samples. Journal
of Microbiological Methods 57: 1-7.
3. Ralf Hartemink and Franklin Georgsson, 1991. Incidence of Listeria species in seafood
and seafood salads. International Journal of Food Microbiology 12(2-3): 189-195.
4. Ronda Dillon, Thakor Patel and Samuel Ratnam, 1994. Occurrence of Listeria in hot and
cold smoked seafood products. International Journal of Food Microbiology 22(1): 73-77
5. Joanne Thimothe, Jonathan Walker, Varanuch Suvanich, Ken L. Gall, Michael W. Moody &
Martin Wiedmann, 2002. Detection of Listeria in crawfish processing plants and in raw,
whole crawfish and processed crawfish (Procambarus spp.). Journal of food protention,
65(11): 1735-1739
6. Victoria R. Lappi, Joanne Thimothe, Johnathon Walker, Jon Bell, Kenneth Gall, Michael W.
Moody and Martin Weidmann, 2004. Impact of intervention strategies on Listeria
contamination patterns in crawfish processing facilities: a longitudinal study. Journal of
food protection 76(6): 1163-1169.
7. Elliot T. Ryser, Samuel M. Arimi and Catherine W. Donnelly, 1997. Effects of pH on
distribution of Listeria ribotypes in corn, hay and grass silage. Appllied & Environmental
Microbiology, 63(9): 3695-3697.
8. Todd J. Pritchard, K.J. Flanders, C.W. Donelly, 1995. Comparison of the incidence of
Listeria on equipment versus environmental sites within dairy processing plants.
International Journal of Food Microbiology, 26: 375-384.
9. Young-Chun Choi, Sun-Young Cho, Boo-kil Park, Duck-Hwa Chung & Deog-Hwan Oh,
2001. Incidence and characterization of Listeria spp. From foods available in korea.
Journal of Food Protection, 64(4): 554-558.
Petrifilm Yeast and Mold Count plates (PYM)

( ) .

Potato dextrose agar nutrients

3MTM Swab kit ()

3M , Swab kit .

3MTM Quick swab

swab
.
1.0mL .
5 .
Letheen broth .
wet swab dry swab .
.
Rayon Ca alginate .

3MTM e-Swab
10ml Buffered Peptone Water (BPW) .
.
10mL .
Rayon Ca alginate .

e.Swab Quick Swab

BPW Letheen brith
No Yes

10 mL 1 mL
Wet & Dry swab
Swab head Rayon Rayon