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Adequacy of in situ glass slides and direct sand extractions to assess the microbiota within sand columns used for drinking water treatment

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Historically, Cholodny-Rossi buried glass slide techniques have been used to study the microbiota of subsurface environments, yet the bias of such a technique has not been compared against direct sand extraction using modern in situ probing. Over a period of 34 wk, four separate 4-m-deep sand columns receiving raw lake water were examined to compare direct extraction of sand filter biofilm material against in situ glass slide biofilms. Significantly different DAPI direct counts and fluorescent in situ hybridization signals for major phylogenetic groups were observed. Not only were lower proportions (P < 0.001) of EUB338-probed DAPI cells observed on in situ glass slides, but also fewer -Proteobacteria (12%–21%) and more α-Proteobacteria (16%–33%) when compared to direct sand extracts. Hence, investigators of the microbial ecology of even simple sand biofilms must consider the inherent biases from "accepted" methods and seek further independent methods to identify those which may be most accurate.Key words: sand filter, biofilms, in situ hybridization, groundwater recharge.

Les techniques de Cholodny-Rossi basées sur des lamelles de verres enfouies ont été utilisées historiquement pour l'étude de la microflore d'environnements en subsurface. Néanmoins, le biais d'une telle technique n'a jamais été comparé et confronté à une extraction de sable directe employant un sondage in situ moderne. Sur une période de 34 semaines, quatre colonnes de sable distinctes de 4 m de profondeur recevant de l'eau de lac brute ont été examinées afin de comparer l'extraction directe de matériel de biofilms de filtres de sable par rapport à des biofilms in situ sur des lamelles de verre. Un nombre significativement différent de comptes directs par DAPI et de signaux par hybridation in situ en fluorescence ont été observés pour les groupes phylogénétiques majeurs. Non seulement de plus faibles proportions (P < 0,001) de cellules DAPI sondées avec EUB338 furent observées sur des lamelles de verre in situ mais, de surcroît, moins de -Proteobacteria (12 %–21 %) et davantage de α-Proteobacteria (16 %–33 %) furent observés par rapport aux extraits de sable directs. Ainsi, les chercheurs s'intéressant à l'écologie microbienne de simples biofilms de sable doivent prendre en compte les biais innés de méthodes « acceptées » et explorer des méthodes alternatives afin de déterminer laquelle serait susceptible de donner une mesure adéquate de la réalité.Mots clés : filtre de sable, biofilms, hybridation in situ, recharge de l'eau souterraine.[Traduit par la Rédaction]

Keywords: biofilms; filtre de sable; groundwater recharge; hybridation in situ; in situ hybridization; recharge de l'eau souterraine; sand filter

Document Type: Research Article

Publication date: 2001-07-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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