-The propionic acid bacterial tlora in Swiss raw milk was investigated. Four hundred and fifty-three strains from lowland milk and 21 strains from Alpine raw milk were c1assified to the species level by protein profile analysis and restriction analysis of the 23S ribosomal ribonucleic acid (rRNA). Plasmid profiles and random amplified polymorphie deoxyribonucleic acid (RAPD) was used to differentiate the strains. The tlora was found to be extremely diverse. Ali four dairy Propionibacterium species were found in lowland raw milk: 71 % were P.freudenreichii, 19 % P.jensenii, 8 % P. acidipropionici and 2 % P. thoenii. P. acidipropionici was not found in alpine milk but P. freudenreichii made up 55 %, P. jensenii 15 % and P. thoenii 30 % of the total. Among 278 P.freudenreichii strains, 219 (79 %) different strains were identified by RAPD to the strain level. For the other species strain diversity was even greater. Only 30 % of the strains analyzed carried plasmids. The Swiss chee se industry has a large reservoir of Propionibacterium strains in raw milk for future applications and developments. © InralElsevier, Paris propionibacterium / milk / electrophoresis / PCR Résumé -La flore propionique dans le lait cru suisse en provenance de la plaine et des alpages.La flore propionique dans le lait cru suisse a été analysée. L'analyse des profils protéiques et l'analyse de restriction de l'ARNr 235 ont été faites avec 453 souches bactériennes du lait en provenance de la plaine et 21 souches du lait d'alpages, en vue de les classer en espèces. Leur contenu en plasmides et la méthode de l'amplification génique au hasard de l'ADN ont été utilisés pour la diffé-renciation des souches. La tlore propionique dans le lait cru suisse s'est avérée être extrêmement variée. Les quatre espèces laitières de Propionibacterium ont été trouvées dans le lait cru de plaine; 71 % étaient des P. f reudenreichii, 19 % des P. jensenii, 8 % des P. acidipropionici et 2 % des P. thoenii. Dans le lait cru d'alpage P. acidipropionici n'a pas été trouvé, P. freudenreichii formait 55 %, P.jensenii 15 % et P. thoenii 30 % des bactéries propioniques. Parmi les 278 souches de P.freudenreichii isolées 219 (79 %) souches différentes ont été identifiées par RAPD. La diversité chez les autres
-One hundred and thirty-five propionibacteria isolated from brown spots of Appenzell, Emmental, Raclette and Sbrinz cheese were analyzed by random amplified polymorphie deoxyribonucleic acid (RAPD) and protein profiling. Both Propionibacteriumfreudenreichii subsp. shennanii and P.freudenreichii subsp.freudenreichii seem to be involved in the formation ofbrown spots in ail these cheeses. In ail Appenzell cheeses, in addition to P.freudenreichii, slow-growing P. acidipropion ici and P. jensenii strains, which are known to cause brown spots, were found. A high proportion of unknown bacteria were noted in the brown spots of Raclette cheese. None of the isolated strains from Appenzell, Raclette and Sbrinz came from the commercial propionibacteria used in cheese production. Only one of the strains isolated was found in raw milk obtained from the same region where the cheese was produced. © Inra/Elsevier, Paris Propionibacterium / cheese / electrophoresis / PCR Résumé -Identification des bactéries propioniques dans les points bruns des fromages suisses à pâte dure et mi-dure. Cent trente cinq bactéries propioniques isolées des points bruns des fromages emmental, sbrinz, appenzell et raclette ont été analysées par RAPD et par SDS-PAGE de leurs protéines. Diverses souches des deux sous-espèces de P. freudenreichii, shermanii etfreudenreichii, semblent jouer un rôle dans la formation des points bruns dans ces fromages. Non seulement des souches de P. freudenreichii mais également de souches à croissance lente de P. acidipropionici et P. jensenii, qui sont connues pour favoriser la formation de points bruns, ont été trouvées dans tous les fromages d'appenzelI. Une proportion importante de bactéries inconnues ont été trouvées dans les points bruns des fromages à raclette. Aucune des souches isolées des fromages sbrinz, appenzell et raclette ne provenait des cultures commerciales fournies par la station fédérale de recherches laitières suisse. Parmi les souches isolées, une seule a aussi été trouvée dans le lait cru obtenu de la region où le fromage avait été produit.
-An amplified sequence of approximately 350 bases from the 23S rRNA gene of each bacteria was used to identify 37 strains of dairy propionibacteria. After generation of the fragment by PCR (polymerase chain reaction), it was eut with the restriction endonuclease MspI. The resulting electrophoretic pattern showed a distinct profile for each of the Propionibacterium species. Purification of DNA was not necessary and the analyses could also be performed with crude bacterial extract. A collection of anaerobie bacteria commonly found in milk and milk products was also submitted to the analysis. None yielded the same pattern as propionibacteria.The analysis of the propionibacteria was compared to that performed by electrophoresis of the soluble cell-free protein extracts and that of c1assical microbiological tests. Our method gave identical results and was less time consuming (results could be obtained within a single day and were also easier to read) than both the c1assical tests and protein electrophoresis. © Inra/Elsevier, Paris.
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