RESUMEN
OBJECTIVES: To identify the genes responsible for tetracycline resistance in a strain of Streptococcus australis isolated from pooled saliva from healthy volunteers in France. S. australis is a viridans Streptococcus, originally isolated from the oral cavity of children in Australia, and subsequently reported in the lungs of cystic fibrosis patients and as a cause of invasive disease in an elderly patient. METHODS: Agar containing 2 mg/L tetracycline was used for the isolation of tetracycline-resistant organisms. A genomic library in Escherichia coli was used to isolate the tetracycline resistance determinant. In-frame deletions and chromosomal repair were used to confirm function. Antibiotic susceptibility was determined by agar dilution and disc diffusion assay. RESULTS: The tetracycline resistance determinant from S. australis FRStet12 was isolated from a genomic library in E. coli and DNA sequencing showed two open reading frames predicted to encode proteins with similarity to multidrug resistance-type ABC transporters. Both genes were required for tetracycline resistance (to both the naturally occurring and semi-synthetic tetracyclines) and they were designated tetAB(46). CONCLUSIONS: This is the first report of a predicted ABC transporter conferring tetracycline resistance in a member of the oral microbiota.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/genética , Antiportadores/genética , Proteínas Bacterianas/genética , Boca/microbiología , Multimerización de Proteína/genética , Streptococcus/genética , Resistencia a la Tetraciclina/genética , Transportadoras de Casetes de Unión a ATP/química , Adulto , Antiportadores/química , Proteínas Bacterianas/química , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Valor Predictivo de las Pruebas , Proteínas y Péptidos Salivales/química , Proteínas y Péptidos Salivales/genética , Streptococcus/aislamiento & purificaciónRESUMEN
We have analyzed the total metagenomic DNA from both human oral and fecal samples derived from healthy volunteers from six European countries to determine the molecular basis for tetracycline and erythromycin resistance. We have determined that tet(M) and tet(W) are the most prevalent tetracycline resistance genes assayed for in the oral and fecal metagenomes, respectively. Additionally, tet(Q), tet(O), and tet(O/32/O) have been shown to be common. We have also shown that erm(B), erm(V), and erm(E) are common erythromycin resistance genes present in these environments. Further, we have demonstrated the ubiquitous presence of the Tn916 integrase in the oral metagenomes and the Tn4451 and Tn1549 integrase genes within the fecal metagenomes.