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CEN/TR 15215-1:2006

Characterization of sludges - Detection and enumeration of Salmonella spp. in sludges, soils, soil improvers, growing media and biowastes - Part 1: Membrane filtration method for quantitative resuscitation of sub-lethally stressed bacteria (to confirm efficacy of log drop treatment procedures)

Edition date: 2006-01-25
In Force
Available languages: German, French, English
Summary: This part of the CEN Technical Report specifies a membrane filtration procedure for the quantitative resuscitation and enumeration, by culture of individual colonies on chromogenic agar media, of Salmonella spp. including potentially sub-lethally damaged Salmonella spp. in sewage sludges. It may be suitable for other sludges, soils, soil improvers, growing media and biowastes but the user shall validate the method using these materials. The fully defined scope will be determined after the proposed validation trials have been agreed and carried out. NOTE 1 The objective is to cover untreated and treated sludges, soils, soil improvers, growing media and biowastes. The method is particularly suited to determining the efficiency of treatment procedures for the elimination of pathogens in sewage sludge as outlined in the Revision of Directive 86/278/EEC (3rd Draft, CEN/TC 308 – doc 525). Treatment type A processes are initially to be validated through a to be defined Log10 reduction with a test organism such as Salmonella senftenberg W775. The method has a limit of detection of approximately 1 cfu/g wet weight sludge, dependent on the solids content which at high concentrations (> 20 % (w/v)) can restrict filtration of the sample volume through the membrane if not first diluted. NOTE 2 Salmonella spp. can be present in biosolids including untreated and treated sewage sludge as both vegetative and sub-lethally damaged cells; the latter require resuscitation to enable colony growth for accurate enumeration on agar media. NOTE 3 This method is not suitable for treated sludges containing less than 1 viable Salmonella spp. per 1 g wet weight. NOTE 4 This method is not suitable for untreated sludges containing low levels of Salmonella.

Dieses Dokument legt ein Membranfiltrationsverfahren für die Quantifizierung von Salmonella ssp., einschließlich möglicherweise subletal geschädigter Salmonella ssp., in Klärschlamm durch Kultivieren von einzelnen Kolonien auf chromogenen Agarmedien fest. Das Verfahren kann auch für andere Schlämme, Böden, Bodenverbesserungsmittel, Kultursubstrate und Bioabfall geeignet sein, der Anwender muss jedoch das Verfahren unter Verwendung dieser Materialien validieren. Der vollständig definierte Anwendungsbereich wird bestimmt, nachdem die vorgeschlagenen Validierungsuntersuchungen vereinbart und durchgeführt sind. ANMERKUNG 1 Der vorgesehene Anwendungsbereich umfasst die Untersuchung von unbehandelten und behandelten Schlämmen, Böden, Bodenverbesserungsmitteln, Kultursubstraten und Bioabfall umfasst. Das Verfahren ist besonders geeignet zur Bestimmung der Wirksamkeit von Behandlungsverfahren zur Beseitigung von Krankheitserregern im Klärschlamm, die in der Überarbeitung der Richtlinie 86/278/EWG (3. Entwurf, CEN/TC 308 – doc 525), Verfahren zur Behandlung vom Typ A, kurz beschrieben sind und die zunächst durch eine festzulegende log10-Reduzierung mit Prüfkeimen, wie z. B. Salmonella Senft Das Verfahren hat eine Nachweisgrenze von etwa 1 KBE/g Feuchtschlammmasse, in Abhängigkeit vom Feststoffanteil, der bei hohen Konzentrationen (> 20 % (Massenvolumenkonzentration)) und ohne vorherige Verdünnung die Filtration des Probenvolumens durch die Membran behindern kann. ANMERKUNG 2 Salmonella spp. kann in Biofeststoffen, einschließlich unbehandelten und behandelten Klärschlamms, sowohl in Form von vegetativen als auch von subletal geschädigten Zellen vorliegen; letzteres erfordert eine Wiederbelebung, um zur genauen Zählung auf Agarmedien ein Koloniewachstum zu ermöglichen. ANMERKUNG 3 Dieses Verfahren ist nicht für behandelte Schlämme geeignet, die weniger als 1 lebensfähige Salmonella spp. je Gramm Feuchtmasse enthalten.

La présente partie du Rapport Technique CEN définit une méthode par filtration sur membrane permettant la revivification quantitative et le dénombrement, au moyen de la culture de colonies individuelles sur milieu chromogène gélosé, de Salmonella spp., y compris des salmonelles potentiellement endommagées, mais de façon non létale, dans des boues d’épuration. Cette méthode peut s’avérer convenir à d’autres boues, sols, amendements du sol, supports de culture et biodéchets mais l’utilisateur doit valider la méthode au moyen de ce matériel. Le domaine d’application pleinement défini ne sera établi qu’une fois les essais de validation proposés pris en compte et réalisés. NOTE 1 L’objectif est de couvrir le domaine des boues, sols, amendements du sol, supports de culture et biodéchets bruts et traités. Cette méthode permet surtout de déterminer l’efficacité des procédés de traitement pour l’élimination des pathogènes dans les boues d’épuration comme souligné dans la Révision de la Directive 86/278/CEE (3ème version, CEN/TC 308 – doc. 525). Les procédés de traitement de type A doivent être initialement validés au moyen d’un abattement de log10 à définir, avec un organisme d’essai de type Salmonella senftenberg W775. La méthode a une limite de détection d’environ 1 ufc/g de masse de boue humide, dépendant de la teneur en solides qui peut, à forte concentration (> 20 % (m/v)), limiter la filtration d’un volume d’échantillon à travers la membrane si celui-ci n’est pas préalablement dilué. NOTE 2 Salmonella spp. peut être présente dans les biosolides comprenant des boues d’épuration brutes et traitées, que ce soit sous la forme de cellules endommagées végétatives et endommagées de façon non létale, ces dernières nécessitant une revivification pour permettre le développement de la colonie afin d’obtenir un dénombrement précis sur milieu gélosé. NOTE 3 Cette méthode n’est pas adaptée aux boues traitées contenant moins d’une cellule viable de Salmonella spp. par gramme de
ICS: 07.100.99-Other standards related to microbiology
CTN: CEN/TC 308 - 6289

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