RESUMO

Ribosomal operons are great tools for microbe community characterization and for microorganisms relationship study, particularly in the case of the acid lactic bacteria. The ribosomal operon of the probiotic strain Lactobacillus delbrueckii UFV H2b20 was partially characterized. A genomic library of this strain was constructed and the clones with partial ribosomal operon were sub-cloned using the shot-gun method for subsequent sequencing with the forward primer. The sequence analysis revealed that the 3' end of the rDNA 16S was following by the short spacer region 1 (16S-23S) and that the 3' end of the rDNA 23S was following by the short spacer region 2 (23S-5S), which preceded the rDNA 5S. In the flanking region of the rDNA 5S gene of this operon rrn, a region encoding six tRNAs was detected.

Operons ribossomais têm sido instrumentos importantes na caracterização de comunidades microbianas e no estudo de relacionamentos entre microrganismos, principalmente em bactérias do ácido láctico. Operons ribossomais da linhagem probiótica, Lactobacillus delbrueckii UFV H2b20, foram parcialmente caracterizados. Um banco genômico da linhagem foi construído e os clones, contendo parte do operon ribossomal, foram subclonados pelo método de "shot gun", para em seguida serem seqüenciados com primer "forward". As seqüências indicaram a presença da extremidade 3' do rDNA 16S seguida da região espaçadora curta 1 (16S-23S) e a presença da extremidade 3' do rDNA 23S seguido da região espaçadora 2 (23S-5S), que por sua vez precedia o rDNA 5S. Adjacente ao gene rDNA 5S deste operon rrn uma região codificadora de 6 tRNAs foi detectada.

RESUMO

Multidrug efflux mechanisms in bacteria contribute significantly to intrinsic and acquired resistance to antimicrobial agents. Genome analysis have confirmed the broad distribution of these systems in Gram-negative as well as in Gram-positive bacteria. Among resistance mechanisms, the multidrug efflux system or pump deserves special attention, since a cell that has acquired it can simultaneously diminish or even suppress the susceptibility to a wide range of antimicrobials. The efflux system is mediated by transport proteins which confer resistance to toxic compounds. In Gram-negative bacteria, a tripartite efflux system is necessary to expel the drug to the outer medium: a protein localized in the cytoplasmic membrane; another in the periplasmatic space (membrane fusion protein - MFP); and a third in the outer membrane (outer membrane factor - OMF). The drug transport is active, and depends either on the energy provided by ATP hydrolysis or is directly driven by the proton motive force. The transport proteins are grouped in families, according to the homology of the amino acid sequences and to similarity of mechanisms. Among Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa have most of the hitherto identified and studied multidrug efflux systems.

RESUMO

Multidrug efflux mechanisms in bacteria contribute significantly to intrinsic and acquired resistance to antimicrobial agents. Genome analysis have confirmed the broad distribution of these systems in Gram-negative as well as in Gram-positive bacteria. Among resistance mechanisms, the multidrug efflux system or pump deserves special attention, since a cell that has acquired it can simultaneously diminish or even suppress the susceptibility to a wide range of antimicrobials. The efflux system is mediated by transport proteins which confer resistance to toxic compounds. In Gram-negative bacteria, a tripartite efflux system is necessary to expel the drug to the outer medium: a protein localized in the cytoplasmic membrane; another in the periplasmatic space (membrane fusion protein - MFP); and a third in the outer membrane (outer membrane factor - OMF). The drug transport is active, and depends either on the energy provided by ATP hydrolysis or is directly driven by the proton motive force. The transport proteins are grouped in families, according to the homology of the amino acid sequences and to similarity of mechanisms. Among Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa have most of the hitherto identified and studied multidrug efflux systems.

RESUMO

Lactic acid bacteria were isolated in the MRS, D-MRS and M17 modified media, during the processing of Italian salami, obtained from two processing plants, in the State of Paraná. The 484 isolates were tested for their antibacterial activity against Listeria monocytogenes (Microbiology Laboratory-UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) and Escherichia coli (ATCC 25922), by a deferred method. One hundred and fifteen isolates inhibited at least two of the pathogens. The isolates presented larger inhibition against L. monocytogenes. The 45 isolates with antagonistic action on the four indicator bacteria were identified by the system Biolog. Lactobacillus bifermentans prevailed among the isolates identified in MRS medium from the samples of the two processing plants.

Bactérias láticas foram isoladas durante o processamento de salame tipo italiano, obtido a partir de duas plantas de processamento, no Estado do Paraná. Para o isolamento, foram utilizados os meios MRS, D-MRS e M17. Um total de 484 isolados teve sua atividade antibacteriana testada sobre Listeria monocytogenes (Laboratório de Microbiologia - UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) e Escherichia coli (ATCC 25922), utilizando-se o deferred method. Cento e quinze isolados apresentaram zonas de inibição sobre, pelo menos, duas das bactérias indicadoras. Os isolados apresentaram maior inibição sobre Listeria monocytogenes. Os 45 isolados que apresentaram zonas de inibição sobre as quatro bactérias indicadoras foram identificados, utilizando-se o sistema Biolog. Lactobacillus bifermentans predominou entre os isolados identificados, obtidos no meio MRS, a partir das amostras das duas plantas de processamento.

RESUMO

Lactic acid bacteria were isolated in the MRS, D-MRS and M17 modified media, during the processing of Italian salami, obtained from two processing plants, in the State of Paraná. The 484 isolates were tested for their antibacterial activity against Listeria monocytogenes (Microbiology Laboratory-UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) and Escherichia coli (ATCC 25922), by a deferred method. One hundred and fifteen isolates inhibited at least two of the pathogens. The isolates presented larger inhibition against L. monocytogenes. The 45 isolates with antagonistic action on the four indicator bacteria were identified by the system Biolog. Lactobacillus bifermentans prevailed among the isolates identified in MRS medium from the samples of the two processing plants.

Bactérias láticas foram isoladas durante o processamento de salame tipo italiano, obtido a partir de duas plantas de processamento, no Estado do Paraná. Para o isolamento, foram utilizados os meios MRS, D-MRS e M17. Um total de 484 isolados teve sua atividade antibacteriana testada sobre Listeria monocytogenes (Laboratório de Microbiologia - UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) e Escherichia coli (ATCC 25922), utilizando-se o deferred method. Cento e quinze isolados apresentaram zonas de inibição sobre, pelo menos, duas das bactérias indicadoras. Os isolados apresentaram maior inibição sobre Listeria monocytogenes. Os 45 isolados que apresentaram zonas de inibição sobre as quatro bactérias indicadoras foram identificados, utilizando-se o sistema Biolog. Lactobacillus bifermentans predominou entre os isolados identificados, obtidos no meio MRS, a partir das amostras das duas plantas de processamento.