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Monitoring of Bacillus thermodenitrificans OHT-1 in compost by whole cell hybridization

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Abstract:

Thermophilic aerobic composting is a widely practiced method for the disposal of exhaust materials. We isolated a thermophilic bacteria strain from a compost sample under aerobic conditions at 60°C. On the basis of its 16S rRNA sequence and physiological characteristics, this strain was identified as Bacillus thermodenitrificans OHT-1. An 18-subunit oligonucleotide probe for 16S rRNA, labeled with fluorescein isothiocyanate, was developed for the detection of B. thermodenitrificans. Spores and vegetative cells of B. thermodenitrificans OHT-1 were detected in liquid culture and laboratory compost by whole cell hybridization using this oligonucleotide probe. The results obtained by whole cell hybridization were evaluated in growth experiments of B. thermodenitrificans OHT-1 in laboratory compost and were used to enumerate spores and vegetative cells.Key words: compost, Bacillus thermodenitrificans, 16S rRNA, whole cell hybridization.

Le compostage aérobie générateur de chaleur est un procédé largement répandu pour l'élimination et le recyclage des matières organiques. Nous avons isolé une souche de bactérie thermophile à partir d'un échantillon de compost dans des conditions aérobies à 60°C. D'après la séquence de l'ARNr 16S et des propriétés physiologiques, cette souche a été identifiée Bacillus thermodenitrificans OHT-1. Pour détecter ce B. thermodenitrificans, nous avons construit une sonde de 18-mer marquée à l'isothiocyanate de fluorescéine pour détecter l'ARNr 16S. En utilisant cette sonde par hybridation sur des cellules entières, il a été possible de détecter des spores et des cellules végétatives de B. thermodenitrificans OHT-1 en milieu liquide et dans un compost préparé en laboratoire. Les résultats d'hybridation obtenus sur des cellules entières ont été validés par des essais de croissance de B. thermodenitrificans OHT-1 dans un compost de laboratoire et ont permis de dénombrer les spores et les cellules végétatives.Mots clés : compost, Bacillus thermodenitrificans,ARNr 16S, hybridation sur cellules entières.[Traduit par la Rédaction]

Keywords: 16S rRNA; ARNr 16S; Bacillus thermodenitrificans; compost; hybridation sur cellules entières; whole cell hybridization

Document Type: Research Article

Publication date: 2002-09-01

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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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