Actero™ Listeria Enrichment Media

Product Information
INTENDED USE: 4. Constantly stir and heat solution until powder is dissolved (avoid
boiling). The pH should be at 7.0 ± 0.2.
Actero™ Listeria Enrichment Media (“Actero™ Listeria”) is a selective 5. Cool the prepared medium to the appropriate temperature (29, 32
medium, specifically optimized for single-step recovery and enrichment of °C or 35°C) and use immediately.
Listeria spp. from food and environmental samples. Note: improper preparation may result in floating particles or
turbidity in the medium after autoclaving
PRINCIPLE OF OPERATION:
Actero™ Listeria Enrichment Media, “Actero™ Listeria”, combines nutritional Environmental Surface Sample Preparation
components with additional ingredients that are necessary to selectively 1. Use a non-bactericidal sterile cellulose sampling sponge
improve recovery and growth of Listeria. The selective agents present in (8×4×0.3 cm) that has been pre-moistened with D/E buffer.
the Actero™ Listeria have been optimized to efficiently target competing 2. Swab the surface to be tested with one side of the swab(s) in a
normal bacterial flora without affecting the growth of Listeria spp. horizontal direction (approximately 10 cm), and with the other side
in a vertical direction (approximately 10 cm) back and forth (one
CONTENTS: stroke back and one stroke forward) to swab the entire area of
100 cm2.
Dehydrated Actero™ Listeria Enrichment Media is provided in 500 g Bottle 3. Place each surface sampled sponge in a sterile sample bag, and keep
(Cat# FCM-011, BBFCM-011) or in 2 kg Bottle (Cat# FCM-022, at 4 ± 2°C until it is ready for testing. Analyze sample units as soon
BBFCM-022). Each format contains sufficient amount of medium powder as possible after their reception in the laboratory.
to prepare respectively 9.29 L and 37.16 L of liquid medium. 4. When ready to test, pre-warm the prepared Actero™ Listeria at 29, 32
or 35°C depending on the detection method to be used.
ADDITIONAL MATERIALS REQUIRED: 5. Add 90 mL of the pre-warmed Actero™ Listeria to each sponge
All Samples sample in its sample bag.
1. Distilled/deionized water. 6. Homogenize the sample for 30 seconds in a Stomacher® 400
2. Sterile Stomacher® bags or equivalent with and without filter. circulator or equivalent. Hand mixing is an acceptable alternative for
3. Stomacher® or equivalent. stomaching. To hand mix, massage each sponge that is in the sealed
4. Calibrated 10 μL plastic inoculating loops. bag for approximately one minute.
5. Rapid‘L.mono agar plates (RLM).
6. Horse blood overlay agar (HL). Enrichment of Environmental Surface Samples Using Actero™
7. Modified Oxford agar plates (MOX). Listeria Enrichment Media
8. Incubator: at 29 ± 0.5°C, 32 ± 0.5°C and 35 ± 2°C. Actero™ Listeria Culture Method or FoodChek™ Listeria spp.
9. Regular laboratory equipment is also required. assay
Food Contact Surface Samples (Stainless Steel, Plastic)
Environmental Samples Close the bags and incubate the samples in an incubator for 24 h at 29
1. Non-bactericidal sterile cellulose sampling sponges (8×4×0.3 cm) pre- ± 0.5°C or for 28 h at 32 ± 0.5°C.
moistened with neutralizing Dey-Engley buffer (D/E) (FoodChek Cat Non-Food Contact Surface Samples (Ceramic, Sealed Concrete, Rubber)
# FCLS-005). Close the bags and incubate the samples in an incubator for 28 h at 32
± 0.5°C.
PROCEDURE: DuPont™ BAX® System Real-Time PCR Assays for Genus
Actero™ Listeria Enrichment Media Preparation Listeria and for Listeria monocytogenes or Actero™ Listeria
Culture Method
Autoclave Method Environmental Surface Samples (Stainless Steel, Plastic, Sealed Concrete)
1. Prepare a clean bottle for each liter of medium preparation. Close the bags and incubate the samples in an incubator for 20-24 h at
2. Shake container of dry enrichment medium before each use. 35 ± 2°C.
3. Measure 53.8 g of powder into the bottle and add to 1 L of distilled 1. Note: Adherence to temperature is important for accurate results.
water.
4. Constantly stir and heat solution until powder dissolved (avoid Enrichment of Samples Using Actero™ Listeria Enrichment Media
boiling). The pH should be at 7.0 ± 0.2. NOTE: The sample preparation depends on the type and size of the
5. Sterilize the bottle of prepared medium by autoclaving at 110°C for 15 sample. Thus, the protocol to prepare the sample should be chosen in
min. function of those conditions. However, all the other steps are the same.
6. Cool bottle to room temperature and store at 2–8°C for up to 45 days.
Keep away from light. Food Sample Preparation
Liquid Whole Eggs (25 g)
Non-Autoclave Method Actero™ Listeria Culture Method or FoodChek™ Listeria spp. assay
1. Prepare a sterile and clean bottle for each liter of medium 1. Add 150 mL of the pre-warmed (32°C) Actero™ Listeria
preparation. Enrichment Media to each 25 g sample in a non-filtered
2. Shake container of dry enrichment medium before each use. Stomacher® sample bag or equivalent.
3. Measure 53.8 g of powder into the bottle and add to 1 L of sterile 2. Homogenize the sample for 30 seconds at 90 rpm in a
distilled water. Stomacher® 400 circulator or equivalent.

FoodChek™ - Actero™ Listeria Enrichment Media Package Insert – FCM-011, 022, 023, 191, 192, 199, 202, – February 2017 Page 1 of 5
Summary of Enrichment and Analysis of Samples Using Actero™ Listeria Enrichment Media
Sample Type Sample Preparation Analysis of Enriched Samples

Environmental Sample

1. Swab a 100 cm2 surface with a sponge (pre-moistened with D/E buffer) and keep
it in a sterile bag at 4°C until tested.
 By FoodChek™ Listeria spp. Assay
2. Homogenize sponge with 90 mL pre-warmed (29°C or 32°C) Actero™ Listeria
 By Actero™ Listeria Culture Method
Enrichment Media.
Food Contact Surface 3. Incubate at 29 ± 0.5°C for 24 h or at 32 ± 0.5°C for 28 h.
 Stainless steel
 Plastic 1. Swab a 100 cm2 surface with a sponge (pre-moistened with D/E buffer) and keep
it in a sterile bag at 4°C until tested.  By DuPont™ BAX® System Real-Time PCR
2. Homogenize sponge with 90 mL pre-warmed (35°C) Actero™ Listeria Assays for Genus Listeria and L. monocytogenes
Enrichment Media.  By Actero™ Listeria Culture Method
3. Incubate at 35 ± 2°C for 20-24 h.

Non Food Contact 1. Swab a 100 cm2 surface with a sponge and keep it in a sterile bag at 4°C until
Surface tested.
 By FoodChek™ Listeria spp. Assay
 Rubber 2. Homogenize sponge with 90 mL pre-warmed (32°C) Actero™ Listeria
 By Actero™ Listeria Culture Method
 Sealed concrete Enrichment Media.
 Ceramic 3. Incubate at 32 ± 0.5°C for 28 h.

1. Swab a 100 cm2 surface with a sponge and keep it in a sterile bag at 4°C until
Non Food Contact
tested.  By DuPont™ BAX® System Real-Time PCR
Surface
2. Homogenize sponge with 90 mL pre-warmed (35°C) Actero™ Listeria Assays for genus Listeria and L. monocytogenes
Enrichment Media.  By Actero™ Listeria Culture Method
 Sealed concrete
3. Incubate at 35 ± 2°C for 20-24 h.

Food Sample

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (32°C) Actero™

 By FoodChek™ Listeria spp. Assay
 Liquid whole eggs Listeria Enrichment Media.
 By Actero™ Listeria Culture Method
2. Incubate at 32 ± 0.5°C for 28 h.

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (29°C) Actero™

 By FoodChek™ Listeria spp. Assay
Listeria Enrichment Media.
 By Actero™ Listeria Culture Method
2. Incubate at 29 ± 0.5°C for 28 h.
 Cold smoked salmon
1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 22-24 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (32°C) Actero™

 By FoodChek™ Listeria spp. Assay
Listeria Enrichment Media.
 By Actero™ Listeria Culture Method
2. Incubate at 32 ± 0.5°C for 28 h.
 Frozen cooked
shrimp
1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 22-24 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 25 g sample with 100 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Raw frozen salmon Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 18-22 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (32°C) Actero™

 Soft fresh cheese  By FoodChek™ Listeria spp. Assay
Listeria Enrichment Media.
(Ricotta)  By Actero™ Listeria Culture Method
2. Incubate at 32 ± 0.5°C for 28 h.

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Mexican-style cheese Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 22-24 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Ice cream Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 18-22 h.  By Actero™ Listeria Culture Method

FoodChek™ - Actero™ Listeria Enrichment Media Package Insert – FCM-011, 022, 023, 191, 192, 199, 202, – February 2017 Page 2 of 5
 Sample Type Sample Preparation Analysis of Enriched Samples

1. Homogenize (30 sec) 25 g sample with 100 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 0.5°C for 22-24 h.  By Actero™ Listeria Culture Method
 Pasteurized milk
1. Homogenize (30 sec) 25 g sample with 150 mL pre-warmed (32°C) Actero™
Listeria Enrichment Media.  By Actero™ Listeria Culture Method
2. Incubate at 32 ± 0.5°C for 28 h.

1. Homogenize (30 s) 25 g sample with 150 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Bagged fresh spinach Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 22-24 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 125 g sample with 750 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Hot dog sausage Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 26-28 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 125 g sample with 500 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Smoked turkey breast Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 24-28 h.  By Actero™ Listeria Culture Method

1. Homogenize (30 s) 125 g sample with 500 mL pre-warmed (35°C) Actero™  By DuPont™ BAX® System Real-Time PCR
 Cured ham Listeria Enrichment Media. Assays for Genus Listeria and L. monocytogenes
2. Incubate at 35 ± 2°C for 26-28 h.  By Actero™ Listeria Culture Method

3. For the enrichment phase, close the bags and incubate the samples 2. Homogenize the sample for 30 seconds in a Stomacher® 400
in an incubator for 28 h at 32 ± 0.5°C. Adherence to temperature circulator or equivalent.
is important for accurate results. 3. For the enrichment phase, close the bags and incubate the samples
in an incubator for 22-24 h at 35 ± 2°C.
Cold Smoked Salmon (25 g),
Actero™ Listeria Culture Method or FoodChek™ Listeria spp. assay Raw Frozen Salmon (25 g),
1. Add 150 mL of the pre-warmed (29°C) Actero™ Listeria Enrichment DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria
Media to each 25 g sample in a filtered Stomacher® sample bag or and for Listeria monocytogenes or Actero™ Listeria Culture Method
equivalent. 1. Add 100 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment
2. Homogenize the sample for 30 seconds in a Stomacher® 400 Media to each 25 g raw frozen salmon sample in a filtered
circulator or equivalent. Stomacher® sample bag or equivalent.
3. For the enrichment phase, close the bags and incubate the samples 2. Homogenize the sample for 30 seconds in a Stomacher® 400
in an incubator for 28 h at 29 ± 0.5°C. Adherence to temperature circulator or equivalent.
is important for accurate results. 3. For the enrichment phase, close the bags and incubate the samples
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria in an incubator for 18-22 h at 35 ± 2°C.
and for Listeria monocytogenes or Actero™ Listeria Culture Method
1. Add 150 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment Soft Fresh Cheese Ricotta (25 g)
Media to each 25 g cold smoked salmon sample in a filtered Actero™ Listeria Culture Method or FoodChek™ Listeria spp. assay
Stomacher® sample bag or equivalent. 1. Add 150 mL of the pre-warmed (32°C) Actero™ Listeria Enrichment
2. Homogenize the sample for 30 seconds in a Stomacher® 400 Media to each 25 g sample in a filtered Stomacher® sample bag or
circulator or equivalent. equivalent.
3. For the enrichment phase, close the bags and incubate the samples 2. Homogenize the sample for 30 seconds in a Stomacher® 400
in an incubator for 22-24 h at 35 ± 2°C. circulator or equivalent.
3. For the enrichment phase, close the bags and incubate the samples
Frozen Cooked Shrimps (25 g) in an incubator for 28 h at 32 ± 0.5°C. Adherence to temperature
Actero™ Listeria Culture Method or FoodChek™ Listeria spp. assay is important for accurate results.
1. Add 150 mL of the pre-warmed (32°C) Actero™ Listeria Enrichment
Media to each 25 g sample in a filtered Stomacher® sample bag or Mexican-style cheese (25 g), Bagged Fresh Spinach (25 g)
equivalent. DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria
2. Homogenize the sample for 30 seconds in a Stomacher® 400 and for Listeria monocytogenes or Actero™ Listeria Culture Method
circulator or equivalent. 1. Add 150 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment
3. For the enrichment phase, close the bags and incubate the samples Media to each 25 g sample in a filtered Stomacher® sample bag or
in an incubator for 28 h at 32 ± 0.5°C. Adherence to temperature equivalent.
is important for accurate results. 2. Homogenize the sample for 30 seconds in a Stomacher® 400
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria circulator or equivalent.
and for Listeria monocytogenes or Actero™ Listeria Culture Method 3. For the enrichment phase, close the bags and incubate the samples
1. Add 150 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment in an incubator for 22-24 h at 35 ± 2°C.
Media to each 25 g sample in a filtered Stomacher® sample bag or
equivalent.
FoodChek™ - Actero™ Listeria Enrichment Media Package Insert – FCM-011, 022, 023, 191, 192, 199, 202, – February 2017 Page 3 of 5
 Ice Cream (25 g) 3. For the enrichment phase, close the bags and incubate the samples
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria in an incubator for 26-28 h at 35 ± 2°C. Adherence to temperature
and for Listeria monocytogenes or Actero™ Listeria Culture Method is important for accurate results.
1. Add 150 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment
Media to each 25 g sample in a filtered Stomacher® sample bag. Analysis of Enriched Samples
2. Homogenize the sample for 30 seconds in a Stomacher® 400 Actero™ Listeria Culture Method – All samples
circulator or equivalent. 1. At the end of the enrichment phase, streak a loopful of an enriched
3. For the enrichment phase, close the bags and incubate the samples sample over the surface of one MOX and one RLM agar plates.
in an incubator for 18-22 h at 35 ± 2°C. Incubate the streaked plates for 24-48 h at 35 ± 2°C in an
incubator.
Pasteurized Milk (25 g) 2. After the 24 h incubation, examine both the MOX and RLM agar
Actero™ Listeria Culture Method plates for colonies with morphology typical for Listeria spp. with
1. Add 150 mL of the pre-warmed (32°C) Actero™ Listeria Enrichment following re-examination after 48 h of total incubation.
Media to each 25 g sample in a non-filtered Stomacher® sample 3. Use a loop to contact a minimum of 20 (if available) suspect
bag or equivalent. colonies presented in MOX agar plate and collectively streak for
2. Homogenize the sample for 30 seconds in a Stomacher® 400 isolation on one or more HL agar plates. Incubate the streaked HL
circulator or equivalent. plates at 35 ± 2°C for 22 ± 4 h.
3. For the enrichment phase, close the bags and incubate the samples 4. Confirm the presumptive colonies growing on HL agar plates as
in an incubator for 28 h at 32 ± 0.5°C. Adherence to temperature Listeria spp. as recommended in the U.S. Food and Drug
is important for accurate results. Administration Bacteriological Analytical Manual, Chapter 10,
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria Detection and Enumeration of Listeria monocytogenes (2011) or in the
and for Listeria monocytogenes or Actero™ Listeria Culture Method FSIS Microbiology Laboratory Guidebook Chapter 8.09 (2013).
1. Add 100 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment
Media to each 25 g sample in a filtered Stomacher® sample bag or Interpretation and Test Result Report
equivalent. 1. If no suspect colonies with morphology typical of Listeria spp.
2. Homogenize the sample for 30 seconds in a Stomacher® 400 have been found on MOX (typical Listeria spp. are small (≈1 mm),
circulator or equivalent. black, round in shape and are surrounded by a darker zone) and/or
3. For the enrichment phase, close the bags and incubate the samples RLM agar plates (L. monocytogenes colonies will appear blue (pale
in an incubator for 22-24 h at 35 ± 2°C. blue, grey-blue to dark blue) without a yellow halo, L. ivanovii
colonies will appears blue-green with a yellow halo and other
Hot Dog Sausage (125 g) Listeria strains will be white, with or without a yellow halo)
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria following a 48 ± 2 h of total incubation, the sample is considered
and for Listeria monocytogenes or Actero™ Listeria Culture Method as being negative for the presence of Listeria spp.
1. Add 750 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment 2. If the suspect colonies with morphology typical for Listeria spp.
Media to each 125 g sample in a filtered Stomacher® sample bag (see above) have been found on MOX and/or RLM agar plates
or equivalent. after 48 h of total incubation, the sample should be considered as
2. Homogenize the sample for 30 seconds in a Stomacher® 3500 or potentially positive for Listeria spp. The suspect colonies must
equivalent. be confirmed according to the U.S. Food and Drug
3. For the enrichment phase, close the bags and incubate the samples Administration Bacteriological Analytical Manual, Chapter 10,
in an incubator for 26-28 h at 35 ± 2°C. Adherence to temperature Detection and Enumeration of Listeria monocytogenes (2011) or in the
is important for accurate results. FSIS Microbiology Laboratory Guidebook Chapter 8.09 (2013).

Smoked Turkey Breast (125 g)

DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria FoodChek™ Listeria spp. Assay – All samples
and for Listeria monocytogenes or Actero™ Listeria Culture Method Refer to the Analysis section of the package insert for the
1. Add 500 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment FoodChek™ Listeria spp. assay.
Media to each 125 g sample in a filtered Stomacher® sample bag
or equivalent. DuPont™ BAX® System Real-Time PCR Assays for Genus
2. Homogenize the sample for 30 seconds in a Stomacher® 3500 or Listeria and for Listeria monocytogenes – Environmental surface
equivalent. samples
3. For the enrichment phase, close the bags and incubate the samples Refer to the Test Protocol section of the package insert for each
in an incubator for 24-28 h at 35 ± 2°C. Adherence to temperature respective assay.
is important for accurate results.
PRODUCT STORAGE AND SHELF LIFE:
Cured Ham (125 g)
DuPont™ BAX® System Real-Time PCR Assays for Genus Listeria The dehydrated medium should be kept at 12–25°C, in tightly closed bottle
and for Listeria monocytogenes or Actero™ Listeria Culture Method in a dry well ventilated place protected from light. The expiry date is
1. Add 500 mL of the pre-warmed (35°C) Actero™ Listeria Enrichment indicated on the package.
Media to each 125 g sample in a filtered Stomacher® sample bag If the medium has been autoclaved, it can be stored at 2–8°C for up to 45
or equivalent. days, in a tightly closed bottle in a cool dry place protected from light. If
2. Homogenize the sample for 30 seconds in a Stomacher® 3500 or the medium has not been autoclaved, it should be used immediately.
equivalent.

FoodChek™ - Actero™ Listeria Enrichment Media Package Insert – FCM-011, 022, 023, 191, 192, 199, 202, – February 2017 Page 4 of 5
DISPOSAL: delivered hereunder by FoodChek Systems Inc., its affiliates or any other
person on its behalf shall, at the time of delivery, be manufactured to meet
Dispose all materials used and the enrichment medium by autoclaving or FoodChek Systems Inc.’s specifications and all applicable laws. All other
according to approved practices. terms, conditions and warranties, including any warranty of merchantability,
Ensure that all biohazard waste is disposed of according to local, municipal, quality, fitness or suitability for a particular or intended purpose, implied by
provincial, state and/or federal regulations. common law or statue, (implied warranties) are expressly excluded.
PRECAUTIONS: CATALOGUE NUMBER:
Biosafety level 2 procedures should be exercised. FCM-011 (BBFCM-011): Actero™ Listeria Enrichment Media, 500 g
(BMBL, http://www.cdc.gov/biosafety/publications/bmbl5/bmbl.pdf). FCM-022 (BBFCM-022): Actero™ Listeria Enrichment Media, 2 kg
Extreme care should be taken in handling test samples and enrichment
broths. All enrichment broths may contain various pathogens whether they
contain Listeria spp. or not. Moreover, those who are at the highest risk of FOR FURTHER INFORMATION PLEASE CONTACT:
serious illness like pregnant women, and potentially immunocompromised
individuals should have limited access to laboratory rooms or areas where FoodChek Systems Inc.
L. monocytogenes isolation or identification procedures are in progress. Suite 450, 1414 – 8 St. S.W.
Calgary, Alberta, Canada
T2R 1J6
TERMS AND CONDITIONS: Tel: 1-877-298-0208
FoodChek Systems Inc. makes no representations and warranties
concerning its products other than those stated herein. All Product(s)

FoodChek™ - Actero™ Listeria Enrichment Media Package Insert – FCM-011, 022, 023, 191, 192, 199, 202, – February 2017 Page 5 of 5