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, Abstr. 91 st Gen. Meet. Am. Soc. Micro biol, 1991.

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, Antibacterial activity of Lactobacillus sake isolated from dry fermented sausages

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, Surface-associated 1 PrfA-regulated proteins of Listeria monocytogenes synthetized . under stress conditions

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. -sorqvist-s, Heat resistance of Listeria monocytogenes by two recovery media used with and without cold preincubation

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-. P. and M. J. Picketi, , p.362

, Points critiques pour la maftrise (PCM)

, 2. Lors de la commercialisation, p.374

, Pal' aillCUl'ôt il fllll'l()l+e (]LW, danf) celte eilttet.tion upiùéullquo

. Pl, Ofcsslonnels intemdffont. les contl·ôles et val'lficalions auxquol.s ils sont ll'lt'll.t<& en application de l'aJ?tlcJo ll-4 de 1a loi modifi6e du l.er aoiH, p.10

, ~~: ot dë la dh?ectfon de la conctii'l'enc<.~, do la. con~ommalion ct.. de lu J.?épl'esafon de.li r1?audês adJ, demandons quns les direetclll'H dépal'tcmetllaux des set?vlc:nR vrH~l'inah?e

J. :. and !. H. , s alirn(mluires [J>J·oduits laiuc~)·B, pJ?oduils de charct~l.crio, viandes et viand<:S hâchécs, J'll'oduits de satJI'h'lF-Jerle, pât.f~sel'i.es à la ct·èmo lndustl

, Incuber le TSA-YE à 37°C pendant 24 heures ou jusqu'à ce que la croissance soit satisfaisante. · 2. Quand une culture pure est obtenue, effectuer les tests Gram, oxydase et catalase. Seules les bacéries Gram positif, oxydase négative, Sélectionner une colonie suspecte isolée et l'ensemencer en stries sur milieu TSA-YE pour confirmer la pureté

, Mettre en suspension les colonies dans 4,6 ml de solution saline stérile dans un tube à essai. Les bactéries en suspension doivent avoir une turbidité égale au standart n° 1 de Macfarland. Utiliser cet inoculum pour le CAMP test et pour l'inoculation de l'unité de Micro-ID Listeria

, Ouvrir l'emballage métallisé et placer l'unité de Micro-ID Listeria à plat sur la paillasse. ll ne faut pas enlever la bande plastique couvrant les puits

, Ouvrir le couvercle et pipeter approximativement 0,3 nù de la suspension bactérienne dans chaque puits d' inoculation (puits supérieurs)

, Fermer le couvercle et placer la galerie verticalement sur son support. S'assurer que les bactéries en suspension soient en contact avec les disques de substrat et ne pas coucher la galerie ( les disques de détection doivent rester secs)

, Si les deux sont positives, L. monocytogenes est suspecté et la galerie doit être incubée pendant 20 heures supplémentaires. Si ces tests sont négatifs, Après 4 heures, noter les réactions de l'esculine (E) et du rharnnose (RHAM)

, Après 24 heures d'incubation, placer l'unité à plat sur la paillasse, ouvrir le couvercle et ajouter deux gouttes (0, 1 ml à peu près ) de KOH à 20% dans le puits d'inoculation du test de VP uniquement

, Fermer le couvercle et mettre la galerie en position verticale. ll faut être sûr que le KOH a coulé dans la solution du test VP. Ensuite faire tourner l'unité de Micro-ID Listeria de 90° dans le sens des aiguilles d'une montre de telle sorte que les disques dans les cinq premiers puits soient humidifiés

, Remettre la galerie en position verticale et s'assurer par tapotement que la solution n'est pas piégée sous les disques d'identification

, Lire toutes les réactions immédiatement à l'exception du test VP (Laisser la couleur se développer pendant 10 minutes approximativement dans le puits et, ensuite

C. A. , At.kins, Munch-Paterson

, Une souche de Staphylococci; au reus fortement bêta-hémolytique par exemple CIP 5710 (Collection de l'Institut P:lSteur de Paris) Une souche de Rhodococcus equi, par exemple CIP 5869. fi est recomm:mdé de détenir des souches de Lisreria des différentes espèces

, Il est import:mt que le laboratoire reçoive un échantillon réellement représentatif

, Pour la préïJaration de l'échantillon et la prise d'essai, voir la nonne ISO 6887 et les normes concem:mt pour le produit à examiner. ?our b ÇJréparation de la suspension-mère

!. Introduire, ), de façon à obte:ur un rapport prise d'essai 1 milieu d'enrichissement de l/10 (r, x) de prise d'essai (masse ou volun1e) d:J.IJs un volume du prernie:-bouillon d'enrichissc:m:::t. formule Fraser-demi

!. :. P~c, \·cr 0,1 ml du bouillon Fraser-demi incubé obtenu en (5.2.1). i!t maculer 10 ml de bouillon d'enrichissement

, Pré!c·:cr i l'aide d'une anse bouclée un aliquot du bouillon demi-Fraser incubé (5 .2.1) et isoler sur une gélose

, !. c~rtir de l'enrichissement secondaire

, Proc~d.::r comme ci -<lessus à partir du bouillon d'enrichissement formule Fraser incubé 18

, ldcntific::nion présomptive de Listerra son · observauon des géloses d'isolement E:~;d'incubation. ? sur ;;dose séiecuve pour Lisrerra formule Oxford. les !tsrcrra forment en 24 h des colonies grises ou gris-?:~.::-d~:r-..:s !uïS:J.IJtes. d'environ 1 mm de di:::unctre, entourees d'un halo brun-nmr ; après 48 h d'incubation. les -421 -colonies typiques ont un diamètre d'environ 2 mm, sont inci'llStées dans la gélose e: présentent une dépression c,entr;ale. * sur gélose sélective pour

, Sélectionner 5 colonies typiques sur chaque boîte. S'il y en a moins àe 5, les retenir toutes

, L'tm des protocoles cités ci-dessous peut être suivi : ? La technique classique consiste à purifier la colonie puis à l'identifier à l'aide de tests morphologiques

, Une technique alternative consiste à identifier Listeria monocytogenes par la mise en évidcnc:: d'une séqueneo

, Les colonies de Lisreria mesurent environ 1 mm de diamètre, sone tr.mslucides, non pigmentées ; soumises à un éclairage de Henry, elles apparaissent bleuâtres avec une suriace gr

, ex:unen de la mobilité

. L&apos;e, umen de la mobilite est n:::llisë soit au microscope sur un état frais ( 1 ). soit e.'1 milieu gélose

, Préparer un éta.lemcnt d'une su.speru-ion d'une colonie en solution saline :i 0.85o

, Sur w1~ gélose mobilité inoculé: par piqün: cenuale et incubée 48 ha zsoc. lc::s Listt!ria poussent autour de la p1q~e et présentc:m le plus souvent une: pousse rypique dite en ombrelle

, Sélectionner les souches c, J.I.J.Iase (+)

, CAMP/S.a.ureus (+), D-xylose (-), rharnnose( +)

, Milieu d'isolement d'Oxford Curtis

, Milieu de: base: Composition : : · Mélange spécial de peptones, p.23

, Esculine: 1,0 g

, Citrate ferrique ammoniacal : 0,5 g

, Chlorure de lithium : 15,0 g

, Eau : 1 000 ml, p.424

G. D. Artis, M. G. Mitchell, A. F. Gng, and E. J. Griffin, , 1988.

, Préparation : Dissoudre les composants ou le milieu complet déshydratés dans l'eau, en portant à ébullition

, Colistinc sulfate : 20 mg

, Acriflavine HCl : 5 mg

, Cefotetan : 2 mg : Fosfomycine : 10 mg

, Ethanol : 5 ml

, Eau stérile : 5 ml

, Milieu complet : ajouter la quantité de supplément appropriée au milieu de base fondu et refroidi à environ 45 oC, bien homogénéiser

L. , boites ainsi préparées peuvent être stockées quinze jours

, !. ~-1ilieu d'isolement de Mox

, Milieu de base : Composition : Gélose columbia modifiée : 38, vol.1

, Esculine : 1,0 g

, Citrate ferrique ammoniacal : 0,5 g

, Chlorure de lithium : 15,0 g

, Colistine: 0,01 g

, Eau : 1 000 ml

U. Macclain, FSIS method for isolation and identification of Listtri4 monocytogtmes from processed meat and poultry produr.ts, ~boratory communication n? 51, Microbiology Division, Food safecy an> inspection service U.S. Departement of Agriculture, 1989.

P. Milieu-de,

J. , Milieu de base: Composition : Mélange spécial de peptones : 23,00 g

, Amidon de mals: 1,00 g

, Chlorure de sodium : 5,00 g

, Citrate ferrique ammoniacal : 0, vol.50

, Chlorure de lithium : 15,00 g

, Agar agar: 12 à 18,00 g (2)

, Eau : 1 000 ml

P. Van-nettcn, A. Van-de-ven, P. Moseel, and D. A. , A selective and diagnostic medium for use in the enumeration of Listeria son in fonds. lot, J. Food Microbiol, vol.6, pp.187-198, 1988.

, Préparation : Dissoudre des composants de base ou le milieu complet déshydraté dans l'eau en portant à l'ébullition. Si nécessaire, ajuster le pH de sorte qu'après stérilisation il soit de 7 ,2. Répartir le milieu de base gélosé dans des récipients appropriés

, mélange : Acriflavine HCl: 5,0 mg

, Ceftazidime : 20,0 mg

, Sulfate de· Polymyxine B : 10,0 mg

, Soytone : 3,0 g

, Chlorure de lithium : 5,0 g

, Glucose : 2,5 g

, Eau : 1 000 ml

, Conserver entre 0 ct 5 oC, un mois maximum. en évitant toute déshydratation du milieu surtout s'il est coulé en boite de Pétri

, Milieu de base : géiyutc 10 g. -Chlomre de sodium S g

L. , Pour les deux hydrates de carbone, préparer des solutions à S p. 100 en eau distillée. Stériliser par filtration en utilisant une membrane de porosité 0, voir ci-dessous) peuvent être remplacées par des gluci-discs Biomérieux, p.45

&. Examen-des-boites-li and . Suspectes, Test d'illumination d'Henry Pour la source lumineuse, vous pouvez utiliser « Bausch and Lamb Nicholas Illuminator, cat, p.2, 1961.

, S'il n'y .a pas de norme internationale spécifique disponible, il est

. Transférer, 1 ml de l'échantillon pour essai s'il est liquide ou 0,1 ml de la suspension mère pour les autres produits à la surface d'une boite de milieu gébsé (5.3.1) préalablement séché et, vol.0

, Etaler l'inoculum le plus rapidement possible à la surface du milieu gélosé en essayant de ne pas toucher les bords de la boite avec l'étaleur en verre

, Retourner les boites préparées selon 9.2.2 et les faire incuber

, Sélection des boites et interprétation : Après vingt-quatre heures d'incubation, repérer les C<J!onies

. Lignes-direcirices-provisoires-relatives-a-u\-o,

A. Pastel and . Lrises,

, UI: lesquels la matière première a lrubi un tra.i t:arent assainissant est eotuellanent "ir'rlêoelable dena 25 gn ? En ~ les lots de prcxiuits qui se révèlent c:x:ntarn:in3s doivent itre sai sis (et ~letoe!nt après saisie destinés A une utilisaticn particulière tella que fente, Jiu stade de La fabricatiDn : I.e cri:tère d' acc:eptaticn de oes p.tOdui ts p

, Au stade as la dist.ribu.tion : Le critère d 1 acoept:atic:n àe oes prcrluits est filté à titre transitoil:e à 10

, En 1' abserYE oe i:rx.lt tra.i tsrent assainissant, · la QUal.i té sanitaire àe oes prcduits est étroi t:arent liée A la qualité de la ma-t

, Les différentes 91'X1Uêtes rial.is:êes jusqu' è oe jo.rr (plan de surveillarx:la, erx:J.l,lêtes ~fiques dans le cache da l'êpidân:l.e) rocutxent clairement qua oertaines fab:d.catirns peuvent ~tre c:x:rrtaminêes avec une fréQuenoel non négligeable

O. Toutefois and . Cx, et en tout état Cie cause l.ergeaent inférieures A la c:ontam.inaticn ré::essaire t;:lOI.lr ocx::.asià'lner une listériose chez un irrlividu en l:x:Jn état de santé, En cx:nséquerY::a, et o:xrpte tenu des .in:format.icns déjà largeroent diffusées aupris des perscl"liES à risq:ue par les autorités sanitaires, i l a été décidé dans le crrrt:axta ac:tue1 de l' épidanie d 1 swliQuer les 1~ direc:trioes sui ventes lXJ.1t:' les p:rdluits au lait CtU, qui ~ util&ment vous guider dans vos déci s:fCXlS

, Au stade às la fabrication : cri tërel; : " :indécelable d!!ms 25 g" Pour les produits très se.~"'~.sibles (type St Neota:L

, facilement identifiables, quant à l'établissement d'origine et au lot de fabrication, selon les dispositions énoncées ci-aprés. 4.1 -Identification de l'établissement de fabrication Le numéro de code de l'établissement tel qu'il figure dans la liste des usines agréés pour l'exportation vers tous les pays (soit numéro minéralogique du département d'origine suivi d'une ou deux lettres, soit numéro d'agrément = code clair sur -les caisses (carton ru oois) d'expédition -les emballages unitaires de dhaque fromage (soit sur l'étiquette), -soit sur la ooîte elle-même, IV -ETIQUETAGE Les fromages, quelle que soit leur destination, doivent être, à tous les stades de la commercialisation

, Pour les fromages préemballés, le code d'identification reste celui imposé par la réglementation

. Cependant, rien ne s'oppose à ce que le producteur ajoute au code d'usine réglementairement défini un code supplémentaire à usage comrrercial (de 01 à 99) lui permettant de personnaliser ses expéditions par importateur, sur l'emballage unitaire de dhaque fromage (étiquette ou boÎte). Ce code ne devra apparaître ni sur les caisses d'expédition

N. B. , dans le cas des fromages de grande taille destinés à être décrupés, il est recomrrandé de faire figurer sur la "collerette" le nwrero de code de l'établissement de fabrication autant de fois que de parts prévues

D. Le-numéro, identification du lot constitué par le numéro de code de l'établissement de fabrication suivi de la date de conditionnement en quantième de l'année doit être indiqué sur le papier d'emballage des fromages par perforations pour la date, impression ou perforation pour le code usine) ainsi que sur les bulletins d'analyses

, ANTE : r::ans tous les cas, il est trés vivement recOlT\lffindé aux fabricants d'inscrire une date limite d'utilisation optimale (D.L.U.O) sur les emballages afin d'éviter des consommations trop tardives de ces produits. Celle-ci indiquée au noyen de la mention, 1ARQUE IMroRI