Third Conference in Food Microbiology Liège...Third Conference in Food Microbiology 9-10-11...

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All characteristic colonies with Rapid’L.mono® were confirmed as Listeria monocytogenes. From the 159 analysed samples, 68 (42,8%) were positive with at least 1 medium; Among these positive samples, only 55,9% can be detected with the 2 media (ref erence medium and Rapid’L.mono® medium). These 2 media allowed both the detection of 77,9% (53/68) of positive samples. A more precise analysis of the results shows that the Rapid’L.mono® medium is less selective than the reference media. Thus it is not convenient for direct enumeration or for a detection after 24 h preenrichment in s emi-Fraser. However, after 48h enrichment in Fraser, the Rapid’L.mono® allows a more efficient isolation of Listeria monocytogenes i solates among other Listeria species. The addition of an extra plate after an e nrichment of 24h in Fraser allows the detection of more isolates than the incubation of 48h (res ults not shown). ASSESSMENT OF RAPID’L.MONO® MEDIUM FOR THE DETECTION OF LISTERIA MONOCYTOGENES IN NATURALLY CONTAMINATED RAW MEAT AND CHEESE Y. Ghafir 1 , L. De Zutter 2 , G. Vlaemynck 3 , J. Denys 1 , G. Daube 1 * 1 Food microbiology, Faculty of veterinary medicine, University of Liege, Liege, Belgium 2 Food microbiology, Faculty of veterinary medicine, University of Ghent, Ghent, Belgium 3 Kwaliteit van dierlijke producten en transformatietechnologie, Landbouwkundig onderzoek, Melle, Belgium Material and Methods • Forty two cheese, 90 raw meat and 27 dried meat samples were assayed for the presence of Listeria monocytogenes in 25g. For cheese, the ISO-11290 was used with Palcam, Oxford and Rapid’L.mono® as isolation and confirmation media. For meat, the NF-V-08-055 method was used with Palcam and Rapid’L.mono®. For a part of the meat samples, Oxford was used instead of Palcam and an extra plate was inoculated after a 24h incubation in Fraser. Introduction Listeria monocytogenes is an important foodborne pathogen. Cheese and meat are among the most frequently implicated foods. For 10 years, performant detection methods have been developed by the main standardisation organisms. The disadvantage of these methods is that the results based on a few isolates characterisation. The Rapid’L.mono® medium (Sanofi) may be a good alternative for Palcam and Oxford media because it allow to distinguish Listeria monocytogenes among the other Listeria species by using its chromogenic properties. Results and Discussion Table 1. Recovery of Listeria monocytogenes fr om diff erent food with reference media and RapidÕL. mono¨ Third Confer ence in Food Microbiology 9-1 0-11 september 1998 Liè ge Cheese Meat Meat* Dried meat Total Total number of samples 42 63 27 27 159 Number of samples positive with one or both media 5 47 9 7 68 Number of samples positive with both media 4 26 3 5 38 Number of samples positive only with Palcam or Oxford 1 13 1 0 15 Number of samples positive only with Rapid’L.mono® 0 8 5 2 15 Number of samples negative with both media 37 16 18 19 90 *: analysed with the modified NF-V-08-055 method 25 g sample + 225 ml 1/2 Frazer 0,1 ml + 10 m l Fraz er 24 h ± 2 h at 30¡C ± 1¡C 24 h ± 2 h at 37¡C ± 1¡C* Camp-test and biochemical confirmation Isolation Palcam or/and Oxford 24 h ± 2 h at 37¡C ± 1¡C + 18 h - 24 h 24 h ± 2 h at 37¡C ± 1¡C Direct L. monocytogenes Identification Is olation Rapid'L.mono ® 24 h ± 2 h at 37¡C ± 1¡C + 24 h * only for a part of the samples Conclusions The use of Rapid’L.mono® medium is time-saving in order to confirm the presence of Listeria monocytogenes. However, because of its weak selectivity, it should be used after a selective enrichment in Fraser during minimum 24h.

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Page 1: Third Conference in Food Microbiology Liège...Third Conference in Food Microbiology 9-10-11 september 1998 Liège hees eM at* Dri ed meat T otal T otal number of sampl es 4 2 6 3

All characteristic colonies with Rapid’L.mono® were confirmed

as Listeria monocytogenes.

From the 159 analysed samples, 68 (42,8%) were positive with

at least 1 medium;

Among these positive samples, only 55,9% can be detected with

the 2 media (ref erence medium and Rapid’L.mono® medium).

These 2 media allowed both the detection of 77,9% (53/68) of

positive samples.

A more precise analysis of the results shows that the

Rapid’L.mono® medium is less select ive than the reference

media. Thus it is not convenient for direct enumeration or for a

detection after 24 h preenrichment in s emi-Fraser. However,

after 48h enrichment in Fraser, the Rapid’L.mono® allows a

more efficient isolation of Listeria monocytogenes i solates

among other Listeria species.

The addition of an extra plate after an e nrichment of 24h in

Fraser allows the detection of more isolates than the incubation

of 48h (results not shown).

ASSESSMENT OF RAPID’L.MONO® MEDIUM FOR THE DETECTION OF LISTERIA MONOCYTOGENES

IN NATURALLY CONTAMINATED RAW MEAT AND CHEESE

Y. Ghafir1, L. De Zutter2, G. Vlaemynck3, J. Denys1, G. Daube1* 1 Food microbiology, Faculty of veterinary medicine, University of Liege, Liege, Belgium

2 Food microbiology, Faculty of veterinary medicine, University of Ghent, Ghent, Belgium 3 Kwaliteit van dierlijke producten en transformatietechnologie, Landbouwkundig onderzoek, Melle, Belgium

Material and Methods• Forty two cheese, 90 raw meat and 27 dried meat samples were

assayed for the presence of Listeria monocytogenes in 25g. For

cheese, the ISO-11290 was used with Palcam, Oxford and

Rapid’L.mono® as isolation and confirmation media. For meat, the

NF-V-08-055 method was used with Palcam and Rapid’L.mono®.

For a part of the meat samples, Oxford was used instead of Palcam

and an extra plate was inoculated after a 24h incubation in Fraser.

Introduction• Listeria monocytogenes is an important foodborne pathogen. Cheese

and meat are among the most frequently implicated foods. For 10

years, performant detection methods have been developed by the

main standardisation organisms. The disadvantage of these methods

is that the results based on a few isolates characterisation. The

Rapid’L.mono® medium (Sanofi) may be a good alternative for

Palcam and Oxford media because it allow to distinguish Listeria

monocytogenes among the other Listeria species by using its

chromogenic properties.

Results and Discussion

Table 1. Recovery of Listeria monocytogenes fr om diff erent foodwith reference media and RapidÕL. mono¨

Third Confer ence in Food Microbiology9-1 0-11 september 1998

Liè ge

Cheese Meat Meat* Dried

meat

Total

Total number of samples 42 63 27 27 159

Number of samples positive with

one or both media

5 47 9 7 68

Number of samples positive with

both media

4 26 3 5 38

Number of samples positive only

with Palcam or Oxford

1 13 1 0 15

Number of samples positive only

with Rapid’L.mono®

0 8 5 2 15

Number of samples negative with

both media

37 16 18 19 90

*: analysed with the modified NF-V-08-055 method

25 g sample + 225 ml 1/2 Frazer

0,1 ml + 10 ml Frazer

24 h ± 2 h at 30¡C ± 1¡C

24 h ± 2 h at 37¡C ± 1¡C*

Camp-test and biochemical

confirmation

Isolation Palcam

or/and Oxford

24 h ± 2 h at 37¡C ± 1¡C + 18 h - 24 h

24 h ± 2 h at 37¡C ± 1¡C

Direct L. monocytogenes

Identification

Isolation

Rapid'L.mono ®

24 h ± 2 h at 37¡C ± 1¡C + 24 h

* only for a part of the samples

Conclusions• The use of Rapid’L.mono® medium is time-saving in order to

confirm the presence of Listeria monocytogenes. However,

because of its weak selectivity, it should be used after a selective

enrichment in Fraser during minimum 24h.