Revision No.: 00 A.Y.: 2 Semester 2022-2023: Microbiological Test Methods Microbial Analysis of Yeast and Molds

Tarlac State University Villa College of Science
MICROBIOLOGICAL TEST METHODS Food Technology Department
Microbial Analysis of Yeast and Molds
Revision No.: 00 A.Y.: 2nd Semester 2022-2023
Lucinda, San Sebastian,

Tarlac City
Page 1 of 7 Group No.:
APPROVAL PAGE
Written by: Signature: Date: Designation: Rating:

Valencia, Jeane Lennix M.
March 27,
Student
2023
Reviewed and Approved by: Signature: Date: Designation:
March 27,
Christine Louanne B. Alturas Instructor
2023

Tarlac City
A. SCOPE AND PURPOSE
The 3M™ Petrifilm™ Yeast and Mold Count (YM) Plate is a type of rapid testing use as an
indicator that facilitates yeast and mold enumeration, present in dairy products and to assess the
Microbiological Safety of the Products.
B. PRINCIPLE OF METHOD
Food manufacturers frequently test their products for YMC since yeasts and moulds can lead to
food deterioration and decomposition. It can also produce toxins that may lead to potentially serious
illnesses.
The 3M™ Petrifilm™ Yeast and Mold Count Plate is a sample-ready culture medium system
which contains nutrients supplemented with antibiotics, a cold-water-soluble gelling agent, and an
indicator that facilitates yeast and mold enumeration. It identifies food borne yeast and mold in your food
products (Dairy) and in your food processing environment. The rapid Yeast and Mold Count Plate method
is faster and easier than the traditional agar method that requires a five-day incubation period.
C. MATERIALS
EQUIPMENT APPARATUS
Incubator Beaker
Analytical balance Erlenmeyer flask
Biosafety Cabinet Pipette/ Tips
Bag mixer Spreader
Test tube
Test tube holder


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Petrifilm
HDP (Plastic)Bag

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D. PROCEDURE
D.1 Sample Preparation
Start
Prepare 25g of Dairy product then place it in the

HDP bag with 225 mL of BPW.
Mix until homogenize in a bag mixer or you can

manually mix it using you hand.
Ready for testing
End

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Tarlac City
D.2 Inoculation
Control
Start
Place the petri film on a flat and level surface.
Dispense 1 mL of BPW in the center bottom of

the petrifilm
Place the spreader on top to spread evenly

and to not form any bubbles
End

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Sample (25g Dairy and 225mL BPW)
Start
Dispense 1 mL of first dilution of the sample in the

center bottom of the petrifilm
Place the spreader on top of the film to spread the

sample evenly and to not form any bubbles
End
D.3 Incubation
Start
Incubate the sample and the control in the

incubator.
Incubate Yeast and Moulds petrifilm (the control

and the sample) for 5 days at 20-25˚c
Wait until the incubation period is finish
End

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D.4 Interpretation
Start
After incubation, the results are ready for reading/interpretation.
For Yeast count the colonies that has dirty white to brown color.
For Molds, count all the colonies that has blue color.
,
After counting all the colonies decide whether the test is passed or
failed based on the standard given
End
E. REFERENCES
3M Food safety (2018). Yeast and Mold Count Plate

Product instruction
https://www.hettweb.com/applications/fungi-and-yeasts-in-milk-and-dairy-products/

Tarlac City
F. APPENDICES
Microbial Analysis of Enterobacteriaceae

Tarlac City

March 27,
Student
2023


Tarlac City
March 27,
2023
APPROVAL PAGE

Tarlac City
The 3M™ Petrifilm™ Enterobacteriaceae Count plate is a type of rapid testing use to test
the pathogenic bacteria present in the dairy product. It is an effective method in counting the
possible colonies and set to be an indicators of food quality and also for food safety.
Enterobacteriaceae are pathogenic bacteria that can cause illnesses as these bacteria produces
toxins that can be every harmful to human. Enterobacteriaceae have had a large impact on infectious
diseases, clinical microbiology and public health. They are involved in diarrhea diseases and recognize as
one of the major bacterial food-borne pathogens.
 3M™ Petrifilm™ Enterobacteriaceae Count Plates provide a cost-effective, convenient and reliable
method for testing food products, that enable you to quickly determine potential sources of
contamination. It is also used for consistent, fast, automated reading and recording of results as this plate
enumerates all coliforms as well as potential pathogens present in the product being test.
C. MATERIALS
EQUIPMENT APPARATUS
Incubator Beaker
Bag mixer Spreader
Test tube
Test tube holder
Petrifilm
HDP (Plastic)Bag

Tarlac City
D. PROCEDURE
Start


Ready for testing
End

Tarlac City


Tarlac City
D.2 Inoculation
Control Start

the petrifilm
Place the spreader on top to spread evenly and

to not form any bubbles
End
Sample (25g Dairy and 225mL BPW)
Start
Dispense 1 mL of first dilution of the sample in the



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End

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D.3 Incubation
Start

incubator.
Incubate Yeast and Molds petrifilm (the control and

the sample) for 24 hours at 37˚c
End

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D.4 Interpretation
Start
After incubation, the results are ready for reading/interpretation.
Count all the colonies that has red colonies with yellow zone and red
colonies with gas bubbles with/ without yellow zone.
After counting all the colonies decide whether the test is passed or failed
based on the standard given
End
E. REFERENCES
3M Food safety (2018). Enterobacteriaceae Count Plate
Product instruction
https://www.3mphilippines.com.ph/3M/en_PH/p/d/v000530644/

Tarlac City
F. APPENDICES
Microbial Analysis of Lactic Acid Bacteria

Tarlac City
The 3M™ Petrifilm™ Lactic Acid Bacteria Count plate is use to enumerates lactic acid bacteria in
food and environmental samples by appropriate dilutions.
Lactic acid bacteria (LAB) are one of the most significant groups of probiotic organisms,
commonly used in fermented dairy products. Although its good, to much Probiotics, including lactic-acid
producing bacteria, may cause digestive issues and brain fog in some people. They may also negatively
affect immunocompromised people, although this mostly applies to supplements rather than foods.
The 3M™ Petrifilm™ Lactic Acid Bacteria Count Plate is a self-contained, sample ready culture
medium system which contains nutrients, selective agents, a cold-water-soluble gelling agent and a
tetrazolium indicator that facilitates colony enumeration. The plate contains oxygen scavenging
compounds which create an anaerobic environment for the recovery of homo fermentative and hetero
fermentative lactic acid bacteria in the food and beverage industries.
C. MATERIALS
EQUIPMENT APPARATUS
Incubator Beaker
Bag mixer Spreader
Test tube
Test tube holder


Tarlac City
Petrifilm
HDP (Plastic)Bag

Tarlac City
D. PROCEDURE
Start


Ready for testing
End

Tarlac City


Tarlac City
D.2 Inoculation
Control
Start

the petrifilm
Place the spreader on top to spread evenly

and to not form any bubbles
End
Sample (25g Dairy Product and 225mL BPW)
Start
Dispense 1 mL of sixth dilution of the sample in the


End

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Tarlac City
D.3 Incubation
Start

incubator.
Incubate Lactic acid petrifilm (the control and the

sample) for 48 hours at 28-37˚c
End

Tarlac City
D.4 Interpretation
Start
After incubation, the results are ready for

reading/interpretation.
Count all Red colonies regardless of size and Red colonies with gases are called hetero
intensity. Red colonies with or without gasses fermentative, while the colonies without
gases are called homo fermentative
After counting all the colonies decide whether the

test is passed or failed based on the standard
given
End
E. REFERENCES
3M Food safety (2018). Lactic Acid Bacteria Count Plate
Product instruction
https://www.3m.com/3M/en_US/p/d/b00013947/

Tarlac City
F. APPENDICES

Tarlac City
APPROVAL PAGE

March 27,
Student
2023
March 27,
2023

Revision No.: 00 A.Y.: 2 Semester 2022-2023: Microbiological Test Methods Microbial Analysis of Yeast and Molds

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Tarlac State University Villa College of Science

MICROBIOLOGICAL TEST METHODS Food Technology Department

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 1 of 7 Group No.:

Written by: Signature: Date: Designation: Rating:

Reviewed and Approved by: Signature: Date: Designation:

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 2 of 7 Group No.:

A. SCOPE AND PURPOSE

Analytical balance Erlenmeyer flask

Biosafety Cabinet Pipette/ Tips

Bag mixer Spreader

Test tube holder

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 2 of 7 Group No.:

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 3 of 7 Group No.:

D.1 Sample Preparation

Prepare 25g of Dairy product then place it in the

Mix until homogenize in a bag mixer or you can

Ready for testing

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 3 of 7 Group No.:

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 4 of 7 Group No.:

Place the petri film on a flat and level surface.

Dispense 1 mL of BPW in the center bottom of

Place the spreader on top to spread evenly

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 5 of 7 Group No.:

Sample (25g Dairy and 225mL BPW)

Place the petri film on a flat and level surface.

Dispense 1 mL of first dilution of the sample in the

Place the spreader on top of the film to spread the

Incubate the sample and the control in the

Incubate Yeast and Moulds petrifilm (the control

Wait until the incubation period is finish

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 5 of 7 Group No.:

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023

Lucinda, San Sebastian,

Page 6 of 7 Group No.:

After incubation, the results are ready for reading/interpretation.

3M Food safety (2018). Yeast and Mold Count Plate

Microbial Analysis of Yeast and Molds

Revision No.: 00 A.Y.: 2nd Semester 2022-2023