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    Evaluatie van de membraanfiltratiemethode op mCP-agar voor bepaling van sporen van Clostridium perfringens in water

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    Authors
    Schets FM
    Medema GJ
    Series/Report no.
    RIVM Rapport 289202007
    Type
    Report
    Language
    nl
    
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    Title
    Evaluatie van de membraanfiltratiemethode op mCP-agar voor bepaling van sporen van Clostridium perfringens in water
    Translated Title
    Evaluation of the membrane filtration method on mCP-agar for determination of Clostridium perfringens spores in water
    Publiekssamenvatting
    Current Dutch and European drinking water standards include criteria for spores of sulphite reducing clostridia. This has some inherent disadvantages. The reproducibility of the enumeration method for spores of sulphite reducing clostridia (SSRC) in Sulphite Cycloserine Agar (SCA) is poor. Some sulphite reducing clostridia are able to grow in sediments in drinking water mains and not all members of this group are of fecal origin. Enumeration of SSRC gives information about the effectiveness of water purification processes rather than about fecal pollution. Clostridium perfringens, however, is exclusively of fecal origin and does not grow in water or sediments. Besides from being a process indicator, C.perfringens spores could be an index parameter for the occurrence of persistent intestinal pathogens like viruses and (oo)cysts of protozoa. This report describes the evaluation of a membrane filtration method for the detection of C.perfringens spores on mCP-agar. The recovery of C.perfringens WR62 spores on mCP-agar was 63,6 +- 10,2% compared to sheep blood agar ; on SCA it was 73,5 +- 4,8%. The selectivity of mCP-agar is high ; 97,5% of the charateristic colonies isolated from sewage samples were confirmed as C.perfringens. The sensitivity, however, is low ; 8/24 (33,3%) of the non-characteristic colonies were also confirmed as C.perfringens. This implies the confirmation of both characteristic and non-characteristic colonies on mCP-agar. Numbers of SSRC and C.perfringens spores were determinated in ten sewage water samples of domestic, industrial and slaughter-house origin on resp. SCA and mCP-agar. In sewage water samples most of the SSRC are C.perfringens spores. In 8/10 samples no significant differences between SCA and mCP-agar occurred. In these samples the average recovery on mCP-agar compared to SCA was 103,8 +- 24,5%. In 2/2 slaughter-house samples significantly higher numbers SSRC on SCA were found. The assumption that most SSRC in sewage water are C.perfringens spores may not apply to this type of sample. As the isolated SSRC were not determined to the species level, we are unable to pronounce upon this with certainty. The mCP-method, however, can be used to determine the number of C.perfringens spores in sewage water of human origin.
    De methode voor bepaling van sporen van sulfiet reducerende clostridia (SSRC) op Sulfiet Cycloserine Agar (SCA) is slecht reproduceerbaar. Een aantal sulfiet reducerende clostridia groeit na in water of in sediment en niet alle sulfiet reducerende clostridia zijn van faecale oorsprong. Bepaling van het aantal SSRC geeft meer informatie over het functioneren van zuiveringsprocessen dan over faecale verontreiniging. Clostridium perfringens is exclusief van faecale oorsprong en vertoont geen nagroei in water en sediment. Sporen van C.perfringens zouden een index functie kunnen vervullen voor het voorkomen van persistente darmpathogenen zoals virussen en (oo)cysten van protozoa. Dit rapport beschrijft de evaluatie van een membraanfiltratie methode voor de detectie van sporen van C.perfringens op mCP-agar. De gemiddelde recovery van sporen van C.perfringens WR 62 op het mCP-medium bedroeg 63,6 +- 10,2% ten opzichte van schapebloed agar. Op SCA was deze recovery 73,5 +- 4,8%. De selectiviteit van het mCP-medium is hoog ; 97,5% van de karakteristieke kolonies is met behulp van de MNLG-tests als C.perfringens bevestigd. De specifiteit is echter minder goed ; 8/24 (33,3%) niet-karakteristieke kolonies werden toch als C.perfringens bevestigd. Dit heeft tot gevolg dat zowel karakteristieke als niet-karakteristieke kolonies op mCP-agar bevestigd moeten worden. De aantallen SSRC en sporen van C.perfringens werden in 10 monsters (huishoudelijke/industrieel/slachthuis) afvalwater met behulp van resp. de SCA- en de mCP-methode bepaald. In afvalwater bestaan SSRC grotendeels uit sporen van C.perfringens. In 8/10 onderzochte monsters bestond geen significant verschil tussen de SCA- en mCP-methode. De gemiddelde recovery op het mCP-medium ten opzichte van SCA bedroeg voor deze monsters 103,8 +- 24,5%. In 2/2 onderzochte monsters slachthuisafvalwater werd met de SCA-methode een significant hoger aantal SSRC dan sporen van C.perfringens met de mCP-methode gevonden. Mogelijk gaat de aanname dat SSRC in afvalwater grotendeels uit sporen van C.perfringens bestaan voor dit type afvalwater niet op. De geisoleerde SSRC zijn echter niet gedetermineerd waardoor hier niet met zekerheid een uitspraak over gedaan kan worden. De mCP-methode kan echter goed gebruikt worden voor het bepalen van het aantal C.perfrigens sporen in afvalwater van humane herkomst.
    Publisher
    Rijksinstituut voor Volksgezondheid en Milieu RIVM
    URI
    http://hdl.handle.net/10029/9976
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