dnp ﬁlm medium for microorganism testing · after opening fold the end of the bag over twice, and...

TM

DNP film medium for microorganism testing

1 2

Simplifying and Optimizing Food-Related Microorganism Testing

DNP Medi·Ca Colony Counting System • Culture images are captured and colonies are automatically counted in a single step, increasing efficiency and saving time.

• DNP's unique image analysis technology - high colony detection accuracy

• Test results & images are simply traceable

Time & space saving

• No need for medium preparation and sterilization

• Speedy sample inoculation without special equipment

• Stackable design greatly reduces the operation space required

Simple operation, reliable results

• Extensive training is not required

• Test results correlate well with established conventional laboratory procedures (AOAC PTM certified)

• Color universal design (CUD) allows for easy distinction between tests

DNP film medium Medi·Ca for microorganism testing is certified by the Color Universal Design Organization, as made in consideration of color universal design, which can be better visualized by more people regardless of individual differences in color vision.

020406080100120140160180

Agar method Medi·CaCO2 Emission Amount (g-CO2e/sheet)

55%less CO2

DisposalFrom raw material procurement to manufacture

Volume is 1/20th that of a petri dish

Reducing the environmental burden

• Volume is 1/20th that of a petri dish - waste can be reduced

• Greenhouse gas (GHG) emissions are also reduced compared to traditional agar tests

*In-company investigation

3 4

How to Use

Disposal

Interpretation

The use of chromogens assist in the visualization of bacterial colonies and counting.

For a Large Number of ColoniesCount colonies within 1-3 squares printed on the cover and calculate an average.

Multiplying the average number by 20 provides the estimated count.

Colony IsolationLift the cover and pick a single colony from the gel.

1. Add appropriate strile diluent to a food sample and blend it with a blender, stomacher or pulsifier.

2. Dilute the sample solution so that the number of bacteria is within an appropriate range (250 cfu/mLor less).

Storage

Keep in the refrigerator (2-8°C).

After opening

Fold the end of the bag over twice, and seal with a tape.The shelf life under refrigerated conditions is 3 months after opening.

Incubation

Incubate the sheets, in lots of up to 25 sheets.

Type of medium Incubation Color of colonies

Blue

Blue

Red

E. coli: navy-blue/blue-violetNon-E. coli coliform: pink/red-purple

35±1 ℃

35±1 ℃

35±1 ℃

35±1 ℃37±1 ℃

48±2 hours

24±1 hours

24±1 hours

24±1 hours

(For Aerobic Bacteria)AC

(For Coliform Bacteria)CC

(For Staphylococcus aureus)SA

(For E. coli & Coliform)EC

Inoculation

Place the sheet on a flat surface and lift yhe cover.

Drop the cover onto the sample (No need to manually spread the sample).

Leave the sheet on a flat, horizontal surface for 3 minutes or more.

Increase efficiency by stacking Medi・Ca

New sheets can be stacked on top of sheets in which solidification is still occurring (up to 10 sheets). This saves space and time during sample placement.*Do not apply pressure to the incubation area of the dripped product. The liquid may leak if pressure is applied.

Aseptically remove each sheet from the aluminum bag.

Place a 1 mL sample suspension onto the center of the medium.

1 Sample Preparation

2

3

4

5

6

All used sheets should be considered to be contaminated and should be disposed of it in accordance with the disposal standards of local governments and facilities after appropriate sterilization.

5 6

Example (Strains) Example (Foods)

Escherichia coli (NBRC 15034)

Staphylococcus aureus(ATCC 25923)

Proteus mirabilis (NBRC 105697)

Bacillus licheniformis(NBRC 12200)

Bacillus subtilis(NBRC 3134)

Bacillus cereus(NBRC 15305)

Raw ground chicken

Icecream + E. coli

Green tea leaf

Raw bean sprouts

Raw salmon

Chocolate

*Incubation conditions: 35°C, 48 hours (Diluent: phosphate buffered saline)*These colorings are for example only.

● Brightly colored colonies

● Reducing the spreading of colonies

Tips for counting

Characteristics

Plate count ager

Plate count ager

For Aerobic Bacteria

Plate count agar (log cfu/g)

Medi·Ca AC (log cfu/g)

0

2

4

6

8

10

0 2 4 6 8 10

R = 0.97● Superior performance

● AOAC PTM certified

Foods including Bacillus spp.

Mixed Powder

Pseudomonas aeruginosa

(A): 1 cfu

(B): 4 cfu

Colonies of aerobic bacteria exhibit clear red color by redox indicator.

High correlation withthe plate count agarhas been confirmed witha wide variety of foods.

AC

AC

AC

Incubation: 35±1℃, 48±2 hours

(A) Count a spreading colonies as 1cfu.*Do not count thin, microscopic colonies scattered acrossan area where a colony is spread out.

(B) Merged colonies count the areas with dark coloring.In this example, there are 4 cfu.

Colonies turn red, making them easy tovisualize and count, even when there isa lot of food residue or color.

Colonies of Bacillus spp. that are easilyspread on conventional agar media donot spread on this product.

* In-company investigation



Enterobacter cloacae(NBRC 13536)

Klebsiella pneumoniae(NBRC 14940)

Escherichia fergusonii(NBRC 102419)

Citrobacter freundii(NBRC 12681)

Enterobacter aerogenes(NBRC 13534)

Raw ground chicken

Cookie + E. coli

Raw tuna

Raw ground pork

Pasteurized milk + K. pneumoniae

Radish sprouts

*Incubation conditions: 35°C, 24 hours (Diluent: phosphate buffered saline)

*These colorings are for example only.

● Brightly colored colonies at 24 hours

Coloring of the entire area caused by enzymes contained in foods

Characteristics

For Coliform Bacteria

In some unheated foods and dairy products, residual enzymes in the food may cause blue coloring of the entire incubation area.

If the visibility of the colonies is affected, improvement can be expected by further dilution.

Colonies of coliform bacteria exhibit clear blue color by chromogenic substrate.

VRB agar

CC

CCRaw pork ham

7 8

Powdered cheese (diluted 10 times)

Dilution

Powdered cheese (diluted 102 times)

Powdered cheese (diluted 102 times)+ K. pneumoniae

When coliforms are present

Incubation: 35±1°C, 24±1 hour

Identification is easier compared to agarplate media, since it is not necessary toconfirm the presence or absence of airbubbles or the size of colonies.

● Superior performance


High correlation withthe traditional agar methodshas been confirmed witha wide variety of foods.


Medi·Ca CC (log cfu/g)

VRB agar (log cfu/g)

0

2

4

6

8

10

0 2 4 6 8 10

R = 0.93

9 10

● Superior performance

High correlation with the traditional agar methods has been confirmed with a wide variety of foods.



Medi·Ca EC (log cfu/g)

0

2

4

6

8

10

0 2 4 6 8 10VRB agar (log cfu/g)

R = 0.99

non-E. coliColiform

0

2

4

6

8

10

0 2 4 6 8 10

Medi·Ca EC (log cfu/g)

Chromogenic agar (log cfu/g)

R = 1.00E. coli




Citrobacter freundii(NBRC 12681)

Klebsiella pneumoniae(ATCC13883)

Enterobacter cloacae(ATCC 222)

Raw ground chicken

Raw shrimp

Omelet + E. coli

Roast beef + E. coli

Icecream + E. coli

Raw salmon

*Incubation conditions: 35°C, 24 hours (Diluent: phosphate buffered saline)

*These colorings are an example.


Coloring of the entire area caused by enzyme contained in foods

Characteristics

Colonies of E. coli O157:H7

For E. coli & Coliform

Coloring example (E. coli + Enterobacter cloacae)

E. coli O157:H7 ATCC 43895(Serotype O157:H7, verotoxin I- and II-producing strains)

Raw oyster (diluted 10 times)+ E. coli

Raw oyster (diluted 10 times)

EC

The conventional tests using agar and liquid media can be replacedwith one sheet of this product.

Identification is easier compared to other agar media, since it is notnecessary to confirm the presence or absence of air bubbles or thesize of colonies.

In some unheated foods and dairy products, residual enzymes in the food may cause coloring of the entire incubation area.


While most E. coli produce β-glucuronidase, E. coli O157: H7 does not specifically produce β-glucuronidase, and it therefore exhibits a red-violet color similar to that of non-E. coli coliform.

When E. coli is presentWhen E. coli is present

Colonies of E. coli exhibit blue-violet to navy blue color and colonies of non-E. coli coliform exhibit clear pink to red-violet color using 2 types of chromogenic substrates.

Incubation: 35±1°C, 24±1 hour


11 12

Staphylococcus aureus(ATCC 25923)

Staphylococcus aureus(NBRC 100910)

Staphylococcus aureus(NBRC 13276)

*BP agar: Baird-Parker agar MSEY agar: Mannitol salt agar with egg yolk


For Staphylococcus aureus

● High specificity

*Incubation conditions: Medi Ca: 35°C, 24 hours / BP agar and MSEY agar: 35°C, 48 hours (Diluent: Butterfield's phosphate buffer or phosphate buffered saline)

*These colorings are for example only.

Colonies of Staphylococcus aureus exhibit clear blue color by chromogenic substrate.

Incubation: 35 or 37±1°C, 24±1 hour

Staphylococcus aureus(D0152)

Cooked ham + S. aureus

Pasteurised milk + S. aureus Cream puff + S. aureus

Egg sandwich + S. aureus

Coloring of the entire area caused by enzymes contained in foods

Raw Asari clams(diluted 10 times) + S. aureus

Buckwheat flour (diluted 10 times)

Dilution

SA● Superior performance

Medi·Ca SA (log cfu/g)

Mannitol salt agar with egg yolk (log cfu/g)

0

1

2

3

4

5

6

0 1 2 3 4 5 6

R = 0.96

Medi·Ca SA (log cfu/g)

Baird-Parker agar (log cfu/g)

0

1

2

3

4

5

6

0 1 2 3 4 5 6

R = 0.95High correlation with the traditional agar methods has been confirmed with a wide variety of foods.

Bacillus licheniformis(NBRC 12200)

Bacillus cereus(NBRC 13494)

Bacillus cereus(D0068)

*These pictures show S. aureus ATCC25923

Incubation

Media required

Determination method

Egg-yolk reaction negative

Egg-yolk reaction negative

Positive

*These pictures show S. aureus ATCC13565

SA

Black colony + egg-yolk reaction (Clear zone)

Mannitol degradability + egg-yolk reaction(Opaque zone)

48 hours

Blue colony

24 hours

1 sheet 3 or more plates

SA BP agar MSEY agar

BP agar MSEY agar

Identification is easier compared to agarcount media, since it is not necessary toconfirm the egg-yolk reaction.

The incubation time is decreased byhalf to 24 hours, improving the timeto report results.

For Medi·Ca, only one sheet is required while surface plating method requires three or more plates.

Some S. aureus that exhibit negative egg-yolkreactions on conventional agar media can bedetected.

Many bacteria other than S. aureus are inhibited, and even if they develop, the colonies exhibit pink to red-violet coloring, making identification easy.

In some unheated foods, residual enzymes in the food may cause coloring of the entire incubation area.


Characteristics Example (Strains) Example (Foods)



13 14

Sophisticated image analysis technology unique to DNP

Displaying storage folders

A list of test result folders Retrieval screen

Easy management of the test results and images

Since results and images are automatically saved by date and type of medium, it is easy to trace past test history.

In addition, keyword retrieval can be conducted from the retrieval screen.

● Merged colonies ● Colonies containing air bubbles

● In case the color of sample solutions is dark

* The above counting results are an example

* Depending on the type of food specimen and the incubation conditions, there may be a difference with the visually counted values may occur.

After countingBefore counting

After countingBefore counting After countingBefore counting

● Microcolonies

After countingBefore counting

Colony detection is highly accurate and highly correlated with visually counted values.

Colony Counting System for DNP film medium

3. Various additional operations concerning the counting results

Promoting the efficiency of the counting operation

Simple operation through 3 STEPS1. Scan Medi·Ca with the dedicated scanner

2. Choose the medium type and initiate counting

3. Confirm and modify the counting result

1. Successive scanning with Medi・Ca

*Note: The processing speed is that at the time of shipment and may be slower due to software installation, etc.

2. Rapid counting regardless of the number of colonies

Approximately 5 sec. per sheet

Counting can be done in approximately 5 sec. per sheet*regardless of the number of colonies, and not only thecounting results, but also images are automatically saved.

Certain test results can also be obtained regardless of theproficiency of the inspector, and time saving and burdenreduction of the inspector can be expected.

Counting results can bemodified and comments canbe added by mouse operation.

Results can also be outputand printed for daily reports.

System configuration● Laptop● Dedicated application● Dedicated scanner*The applications and scanners are not sold separately.

*Please contact us directly for more details of the laptop and scanner

Max 14 sheets

Up to 14 sheets can be in the scanner for successivescanning with Medi·Ca.

Additional Medi·Ca can also be set during scanning.

Characteristics●Optimizing the counting operation

● Sophisticated image analysis technology unique to DNP

● Easy management of the test results and images

Precautions for use1. This product is intended to measure the numbers of microorganisms in foods and beverages and is not intended for medical laboratory testing and sterilization testing.

2. This product is not verified for all kinds of food, food manufacturing processes, test protocols and strains. Laboratories should verify its performance in-house before use.

3. Do not open the cover film until immediately before inoculation.

4. Do not use this product after its expiration date.5. Do not use this product if any breakage, deformation, discoloration, pollution or contamination is observed.

6. Do not expose this product to ultraviolet or direct daylight.7. Do not press on the cover film immediately after the sample solution is dropped. The sample solution may leak outside the incubation area.

8. If the sample solution leaks outside the incubation area, replace it with a new product and try again.

9. If this product comes into contact with your eyes or mouth, rinse your eyes or mouth immediately with water and seek medical advice.

10. Since there is always a risk of microbial infection when handling this product, take appropriate measures against biohazards under the guidance of an expert.

11. Handle this product etc. that has been in contact with a sample or sample solution, as a potentially infectious material.

Store in a refrigerator (2 to 8°C).

Storage method

Storage method after openingAfter opening the package, fold the open end of the package at least 2 times and tape it. Store in a refrigerator (2 to 8°C) and use within 3 months.

Expiration dateThe expiration date of this product is indicated on the upper part of this product (the date indicated after "EXP" is the expiration date). The expiration date indicated on the package applies before this product has been opened while it is appropriately stored.

Disposal methodSince this product has a risk of second infection after use, dispose of it in accordance with the disposal standards of local governments and facilities after appropriate sterilization.

Scope of warranty responsibilityIf the product is found to be defective, it will be replaced with a new product based on the number of defective units.The user is entirely responsible for judgment and use of the inspection results, and the manufacturer or distributor of this product shall not be responsible for them.

2019. 9"Medi·Ca" is a trademark or registered trademark of Dai Nippon Printing Co., Ltd.

Business Development Department, Global Business Division, Packaging Operations1-1-1, Ichigaya-Kagacho, Shinjuku-ku, Tokyo 162-8001, Japanhttps://www.dnp.co.jp/CGI/inquiry_eng/form.cgi

dnp ﬁlm medium for microorganism testing · after opening fold the end of the bag over twice, and...

Documents