break the “mold” with rapid yeast and mould detection methods

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Break the “Mold” with Rapid Yeast and Mould detection methods


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Yeast & Mold Testing Yeasts & Molds are an important quality indicator in the food and

beverage industry, impacting spoilage and shelf life

― one of the top four indicator organisms

― Estimated to be 14% of total indicator testing volume

Important food segments for Yeast & Mold:

― Cultured dairy, fruits & vegetables, bakery products,

confectionary, RTE meats, nuts, spices, dry goods

Characteristics of Yeast & Molds:

― Grows well in low pH and low water activity

― Requires long incubation time: 5 days (reference method) vs.

most indicator organisms are incubated 24-48 hrs.

― Difficult to interpret (count): Molds grow as a irregular spreading

mass (as opposed to a pinpoint colony like bacteria)

Total Ct.34%

Coliform15%

E. coli 15%

Yeast and Mold14%

Staph aureus6%

EB6%

Other10%

Indicator Tests Worldwide


Reference methods for Yeast & Mold testingFDA-BAM Reference Methods ISO Reference Methods

FDA-BAM, Chapter 18

Spread-plate or Pour-plate

DRBC, DG18 or PDA, 25oC, 5-7 days*

0.1% Peptone water

25-50 g sample, 11 mL/99 mL (Dairy)

Dairy (SMEDP)

Spread-plate

Yeast extract glucose chloramphenicol agar,

PDA or DRBC, 25oC, 5 days*

Butterfield’s phosphate buffered water

ISO 21527-1 -2008 (Aw >0.95)

Spread-plate

Dichloran rose bengal chloramphenicol agar (DRBC)

0.1% Peptone water, 25oC, 5 days

ISO 21527-2 -2008 (Aw <0.95)

Spread-plate

Dichloran 18% glycerol agar (DG18)

0.1% Peptone water, 25oC, 5-7 days

ISO 6611: 2004 (Dairy)

Pour-plate

Yeast extract glucose chloramphenicol agar

25oC, 5 days

* Count plates after 5 days of incubation. If there is no growth at 5 days, re-incubate for another 48 h


Comparison of Techniques

Prepare Sample

Prepare Agar

Pour plates, allow to solidify

1st Dilution:

• Dispense 1mL over

3 plates

• Spread inoculum

on each plate0.3mL 0.3mL 0.4mL

0.1mL

3rd Dilution:

• Make 1:10 dilution of

the first dilution

• Dispense and spread

0.1mL over one plate

Incubate

0.1mL

Prepare Sample

Prepare Agar

1st Dilution:

Inoculate empty

dish

Pour agar, swirl,

allow to solidify

2st Dilution:

Make further 1:10

dilution, inoculate

1mL, pour agar,

allow to solidify

1.0 mL

Incubate

3rd Dilution:

Make further 1:10

dilution, inoculate

1mL, pour agar,

allow to solidify

1.0 mL

1.0 mL

Prepare Sample

1st Dilution:

Inoculate 3M

Petrifilm Plate

2st Dilution:

Make further 1:10

dilution, inoculate

1mL, on 3M

Petrifilm Plate

Incubate

25

3rd Dilution:

Make further 1:10

dilution, inoculate

1mL, pour agar

Spread Plate Process Pour Plate Process 3M Petrifilm Plate Process

2525

2nd Dilution:

• Dispense 0.1mL of first

dilution on one further

plate

• Spread inoculum


Yeast & Mould’s usually take much longer to grow and be counted than other microbiology tests

The group are very different from other microbes we count in traditional microbiology testing

Eukaryotic cells as compared to bacteria that are Prokaryotic

Very different from Bacteria including size

Often have slower metabolic pathways

Utilise different growing conditions

― Aerobic

― Can grow in lower Water Activity than bacteria

― Lower pH

― Often produce antibiotics so can be difficult competitors

Why does Yeast & Mould testing take so long?


Slow growing due to complex physiology


Complex cellular organisation


Total Viable Count

― 48 hrs with most food matrices

― Some newer tests allow this result within 24 hrs

E.coli Testing

― 24 hrs

Enterobactericeae testing

― 24 hrs with VRBGA or Petrifilm

Yeast & Mould Testing

― 5 days

Times of some common Food microbiology tests


Food testing for Yeast & Mould contamination is normally relate to food spoilage and

not food poisoning

Only a few relevant mycology food poisoning cases are seen every year

Commercially it has a huge impact on the value of food and the amount of food

wasted globally

Mould contamination can be a very visual issue

Yeast contamination often causes taste changes in a food sample

Food testing for Yeast & Mould contamination


As Yeast & Moulds are a very large group we often get mixed

colonies on a plate

These can overlap and make reading very difficult

In some cases a 3 day and a 5 day read are needed

Difficult to read


Opening an agar plate can lead to contamination of your lab with mould spores

For weeks after none selective media can be contaminated with small mould colonies

This can result in the need for cleaning with harsh chemicals such as Phenols

In severe cases the lab can need to be fumigated

Adds a lot of inconvenience and expense in the lab

Laboratory Contamination


Description: 3M Petrifilm Yeast & Mold plate with significantly improved time-to-result and interpretation.

Rapid Petrifilm™ Yeast & Mold Plate – Product Summary

Image of plate post-incubation:

colonies indicated by blue color

Key Features & Benefits:

• Yeast & Mold Plate with 48-60 hour incubation time across a wide variety of foods.

• AOAC RI and ISO-AFNOR validated results

• Easier interpretation on Yeast & Mold plate(special technology that keeps mold colonies from spreading out and overlapping)

• New indicator dye technology colors colonies blue and make them appear faster.

• Foam dam added to plate for easier inoculation

• All of the traditional benefits of Petrifilm plates are maintained: simplicity, ease-of-use, shelf life, consistency, eliminates agar prep.


RY&M Petrifilm Advantages

― Shorter incubation times up to 50% faster

― Easier interpretation less time to read the plates

― Single media use High and low water activity on single plates

― Ready to use. No wait to use the product

― Consistent (Especially important with the addition of antibiotic

supplements. When this is missed out the results given are not

accurate)

― Long shelf life 18 months compared to 3-4 weeks

― Smaller footprint 90% space saving

― Less waste 90% less material to autoclave / pay to dispose

― Sealed plates Reduce the risk of laboratory contamination

Comparison between traditional agar & Rapid Y&M plates


Thanks for your attention

Louise Mare

Technical Manager 3M South Africa

[email protected]

break the “mold” with rapid yeast and mould detection methods

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