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Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage.

ABSTRACT: The likelihood that products prepared from raw meat and milk may act as vehicles for antibiotic-resistant bacteria is currently of great concern in food safety issues. In this study, a collection of 94 tetracycline-resistant (Tc(r)) lactic acid bacteria recovered from nine different fermented dry sausage types were subjected to a polyphasic molecular study with the aim of characterizing the host organisms and the tet genes, conferring tetracycline resistance, that they carry. With the (GTG)(5)-PCR DNA fingerprinting technique, the Tc(r) lactic acid bacterial isolates were identified as Lactobacillus plantarum, L. sakei subsp. carnosus, L. sakei subsp. sakei, L. curvatus, and L. alimentarius and typed to the intraspecies level. For a selection of 24 Tc(r) lactic acid bacterial isolates displaying unique (GTG)(5)-PCR fingerprints, tet genes were determined by means of PCR, and only tet(M) was detected. Restriction enzyme analysis with AccI and ScaI revealed two different tet(M) allele types. This grouping was confirmed by partial sequencing of the tet(M) open reading frame, which indicated that the two allele types displayed high sequence similarities (>99.6%) with tet(M) genes previously reported in Staphylococcus aureus MRSA 101 and in Neisseria meningitidis, respectively. Southern hybridization with plasmid profiles revealed that the isolates contained tet(M)-carrying plasmids. In addition to the tet(M) gene, one isolate also contained an erm(B) gene on a different plasmid from the one encoding the tetracycline resistance. Furthermore, it was also shown by PCR that the tet(M) genes were not located on transposons of the Tn916/Tn1545 family. To our knowledge, this is the first detailed molecular study demonstrating that taxonomically and genotypically diverse Lactobacillus strains from different types of fermented meat products can be a host for plasmid-borne tet genes.

SUBMITTER: Gevers D

PROVIDER: S-EPMC143591 | biostudies-literature | 2003 Feb

REPOSITORIES: biostudies-literature

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Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage.

Gevers Dirk D Danielsen Morten M Huys Geert G Huys Geert G Swings Jean J

Applied and environmental microbiology 20030201 2

The likelihood that products prepared from raw meat and milk may act as vehicles for antibiotic-resistant bacteria is currently of great concern in food safety issues. In this study, a collection of 94 tetracycline-resistant (Tc(r)) lactic acid bacteria recovered from nine different fermented dry sausage types were subjected to a polyphasic molecular study with the aim of characterizing the host organisms and the tet genes, conferring tetracycline resistance, that they carry. With the (GTG)(5)-P ...[more]

PMID: 12571056

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Project description:PurposeFive Lactobacillus strains isolated from sucuk (Turkish dry-fermented sausage) were studied for their genetic and technological properties.MethodsFor genotypic identification, strains 16S rRNA gene sequences were used. To determine the antimicrobial activity of strains, seven foodborne pathogens were tested. Strains technological properties were characterized.ResultsThese strains were identified as Lactobacillus plantarum by 16S rRNA gene sequence analysis and the phylogenetic tree obtained by neighbor-joining method allowed grouping of these strains into three subgroups. L. plantarum strains showed antagonistic activities against Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Micrococcus luteus strains. PCR assay, using specific primers, showed the presence of bacteriocin (plantaricin) encoding genes in all L. plantarum strains tested. Antimicrobial metabolite production of these strains started at log phase and reached the maximum level at the end of the stationary phase. Regarding their technological properties, better growth was observed at 25 °C compared with 15 °C and 45 °C. The isolates which grown well within the pH scale pH 4.5-6.5 range additionally showed a decent growth at 6.5% salt concentration. It has been found that strains do not exhibit lipolytic and proteolytic activities nor have lysine, ornithine, and arginine decarboxylase activity. On the other hand, one strain showed weak nitrate reductase activity, and four strains produced acetoin from glucose. In addition, all strains were DL-lactic acid producers. Consequently, L. plantarum strains isolated exhibited some biochemical properties required for a starter culture in sucuk and similar products.ConclusionsAll identified strains may be a protective culture in the production of fermented meat products. In particular, L. plantarum S51 was distinguished from other isolates due to the inability to form acetoin from glucose. Further work will be needed to characterize L. plantarum strains as starter culture.

Dataset Information

Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage.

Publications

Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage.

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