Detection, enumeration
and confirmation of
Listeria spp. &
L. monocytogenes
Screening of Listeria spp.
and Listeria monocytogenes
Enumeration of
Listeria monocytogenes
Confirmation of
Listeria monocytogenes
ALOA®
Listeria spp.
confirmation
ALOA®
ALOA® first isolation medium
according to EN ISO 11290-1
and 11290-2
Experience
cannot be copied!
ISO 16140
Validation
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ALOA ISO 11290.
ALOA® is a medium used for the screening of Listeria spp. and Listeria monocyto-
genes and enumeration of L. monocytogenes in food and environmental samples
® ®
( and protocols).
Simple protocol: 1 bag - 1 plate
R elease of negative samples
®
after 24 hours incubation on
FDA BAM ALOA® has become the mandatory medium of the ISO protocol
ISO (EN ISO 11290-1 and ISO 11290-2) for the screening and the enu-
AFNOR meration of L. monocytogenes.
CFSAN Listeria grow as blue-green regular round colonies (detection of
β-glucosidase by using a specific chromogenic substrate with ou
NORDVAL without opaque halo).
AOAC pending
Listeria monocytogenes show an opaque halo; this helps to
easily differentiate them from other species of Listeria. The
halo is due to the activity of a phospholipase involved in the
infection process of pathogenic Listeria.
Listeria spp. ®
and L. monocytogenes detection
ISO 16140 validation for food and animal feeding and environmental
samples
ISO 16140 Valid
Incubation
ation
# AES 10/3-09/
24 ± 2 h 00
at 30 °C ± 1 °C
Spread or Streak
ALOA plate with
X g or X ml of sample 0,1 ml
+ 9 X of Fraser 1/2
(Incubation during
48 hours is possible
Incubation
for your convenience; 24 h
i.e.: week-ends) at 37 °C ± 1 °C
Presumption* Absence of Listeria spp Presumption* of
of Listeria spp. and Listeria monocytogenes Listeria monocytogenes
Listeria spp. Confirmation :
Confirmation ®
or standard tests
* According to AFNOR certification, all positive samples have to be confirmed
®
Enumeration of Listeria monocytogenes
ISO 16140 validation for all food and animal feeding samples
ISO 16140 Valid
ation
Revitalisation # AES 10/3-09/
1 h ± 5 min 00
at 20 °C ± 2 °C
X g or X ml of sample + 9 X
of buffered peptone water
S pread plate P our plate
Spread: Pour 1 ml in about 15 ml ALOA® agar cooled at
• F or a result < 100 CFU/g: 47 °C ± 2 °C (for a result < 10 CFU/g)
0,1 ml on plate ALOA® Ø 90 mm
• For a result < 10 CFU/g:
1 ml on plate ALOA® Ø 140 mm
or 1 ml on 3 plates ALOA® Ø 90 mm
Incubation
48 h Confirmation :
®
at 37 °C ± 1 °C
or standard tests
• Absence of typical colonies • Presence of typical colonies
<10 or 100 CFU/g presumption of Listeria monocytogenes
®
Confirmation of
Listeria monocytogenes
®
is the confirmation method
Listeria monocytogenes
of Listeria monocytogenes after detection by
® ® (streak blue/green
or enumeration by + yellow coloration
+ opaque halo)
Confirmation from
a single typical colony
Inoculation by single streak
Up to 6 confirmations per plate
Bacillus cereus
Results within 24 hours only (blue streak + intense Listeria ivanovii
opaque halo, without (blue streak + opaque
yellow confirmation) halo, without yellow
Any positivesample detected by the coloration)
®
, method, then confirmed with
®
do not need to be confirmed
after enumeration.
Listeria spp. confirmation
AES CHEMUNEX introduces 2 confirmatory tests for Listeria spp.
®
confirmation with method.
5 hour protocol
Incubation
5 h at 30 °C ± 1 °C
Subculture one Listeria spp. presumptive (incubation can be exten- Use the Listeria spp. confirmation strip,
colony in 1 ml of BHI broth ded up to 24 hours) 10 to 20 minutes
2 4 hour protocol
Incubation 24 hours at 37 °C
ost effective method :
C
up to 12 confirmations per plate
Confirmation from one single
typical colony
Inoculation by prick Listeria spp.
(greenish colony with
black halo)
Ordering Information
® ®
Broths
AEB620088: ALOA Count® • Bottles packaging: • Dehydrated packaging
AEB520080: 20 plates Ø 90 mm
(bases + supplements) AEB610419L: 1/2 Fraser, 6 bottles 900 ml - Half Fraser
AEB520079: 120 plates Ø 90 mm
AEB120082: 10 plates Ø 140 mm 5 x 180 ml base + supplements AEB610418: ½ Fraser , 6 x 225 ml AEB140412: 1/2 Fraser, 500 g
AEB720088: ALOA One Day® - Dehydrated kit AEB610309L: BPW, 6 x 900 ml AEB140413: 1/2 Fraser, 5 kg
Confirmations AEB610308 : BPW, 6 x 225 ml AEB140417: 1/2 Fraser, 10 kg
© Masson Communication 2010 • DA176C
for 20 analysis (ALOA® plates + dehydrated
• Listeria monocytogenes :
1/2 Fraser) AEB110425S: s upplement for Fraser,
AEB120100 : 10 plates ALOA Confirmation®
AEB720089: ALOA One Day® - Dilubag® kit • Pouches packaging: 5 tubes of 10 ml
90 mm
120 analysis (ALOA® plates + 1/2 Fraser in AEB910303/4: BPW,
Dilubag®) • Listeria spp. : Dilubag® 3 liters x 4 - Buffered Peptone Water
AEB720090: ALOA One Day® - Bottles kit for AEB110101 : 20 tubes BHI 1 ml AEB910305/2: BPW, AEB240309: QS for 9 liters
120 analysis (ALOA® plates + 1/2 Fraser in AEB400050 : listeria spp confirmation strip AEB140302 : 500g
Dilubag® 5 liters x 2
bottles) AEB522050 : 20 plates PALCAM 90 mm AEB910913/4: 1/2 Fraser, AEB140303 : 5 kg
Dilubag® 3 liters x 4 AEB140307 : 10 kg
AEB910915/2: 1/2 Fraser,
Dilubag® 5 liters x 2
ISO 9001: v 2008
[Link] LRQA FAQ 9914921
AES Chemunex AES Chemunex AES Chemunex AES Chemunex AES Chemunex AES Chemunex AES Chemunex
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