Identification of Clostridium tyrobutyricum and related species using sugar fermentation, organic acid formation and DNA probes based on specific 16S rRNA sequences
1994
Klijn, N. | Bovie, C. | Dommes, J. | Hoolwerf, J.D. | Waals, C.B. van der | Weerkamp, A.H. | Nieuwehhof, F.F.J.
Spores of Clostridium tyrobityricum are considered as the causative agents for butyric acid fermentation (late blowing) in brine-salted, semi-hard and hard cheeses. In order to enable the early and specific detection of these bacteria, clostridia commonly present in raw milk and farm environments were classified using phenotypic and genotypic properties. In addition specific DNA probes were designed for the identification of clostridia commonly present in these environments. Phenotypic characterization based on the API 20A system, the formation and conversion of organic acids and the litmus milk reaction, only allowed the differentiation of 71 clostridial strains into three groups, comprising the species C. tyrobutyricum (group A), C. acetobutylicum, C. beijerinckii and C. butyricum (group B) and C. sporogenes and C. bifermentans (group C) respectively. Strains belonging to the C. tyrobutyricum group formed a well-defined and homogenous collection. Using DNA probes based on specific 16S rRNA sequences and DNA-DNA hybridizations a perfect correlation was obtained with the phenotypic classification. The 16S rRNA probes allowed the specific identification of this species. Using specific 16S rRNA probes, the species in phenotypic group B could be readily differentiated from each other and from other clostridia. A 16S rRNA probe was also developed for C. sporogenes belonging to group C. The probe allowed the differentiation of this species from all other clostridia. The identification based on the probes was fully compatible with the results from DNA-DNA hybridization studies.
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