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Six strains (Bi11, Bi30, Bi36, Bi50, Bi52 and Bi55) isolated from bio-yoghurts and two strains (KD10 and KD11) derived from human faeces were identified by genus- and species specific polymerase chain reaction (PCR) with reference to the type strains of B. animalis subsp. lactis DSM 10140 and B. animalis subsp. animalis DSM 20104. The isolates were differentiated by using Bcu I ( Spe I), Xba I and Dra I endonucleases for subsequent pulsed field gel electrophoresis (PFGE) technique and by API 50 CHL tests.All the isolates tested were classified to B. animalis subsp. lactis species. The reliable identification as B. animalis subsp. lactis (by PCR with Bflact2/Bflact5 primers), however, required confirmation by a negative result of B. animalis subsp. animalis -specific PCR.Differentiation of the B. animalis subsp. lactis isolates with PFGE method enabled to distinguish KD11 strain with all the restriction enzymes applied, and Bi11 and Bi30 — exclusively with Dra I and Spe I enzymes, respectively. The biochemical tests, however, revealed that all the strains tested were characterised by a unique fermentation pattern. It was concluded that differentiation of the B. animalis subsp. lactis strains should be carried out on the basis of both genetic and phenotypic features.

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Acta Alimentaria
Authors: J. Krulj, N. Ćurčıć, A. Bočarov Stančıć, J. Kojıć, L. Pezo, L. Peıć Tukuljac, and M. Bodroža Solarov

programs at NCBI database. 1.3 Restriction enzyme digestion The PCR products, amplified by the primers ITS1/ITS4, were digested with the Hha I and Mwo I (HpyF10VI) enzymes (Thermo Scientific, USA). Digestion was performed by incubating a 10 μl aliquot of

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Dlauchy, D., TOrnai-Lehoczki, J. & Péter, G. (1999): Restriction enzyme analysis of PCR amplified rRNA as a taxonomic tool in yeast identification. Syst. appl. Microbiol. , 27 , 445-453. Restriction enzyme analysis of PCR

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.H.J. & van Der Vossen, J.M.B.M. (1995): Random amplified polymorphic DNA and restriction enzyme analysis of PCR amplified rDNA in taxonomy: two identification techniques for food-borne yeasts. J. appl. Bacteriol. , 79 , 525

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.L. (1987): Restriction enzyme analysis of lactose and bacteriocin plasmids from Streptococcus lactis subsp. diacetylactis WM4 and cloning of Bc/I fragments coding for bacteriocin production. Appl. environ. Microbiol. , 53 , 1171

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