RESUMO
Cotrimoxazole, the combined formulation of sulfamethoxazole and trimethoprim, is one of the treatments of choice for several infectious diseases, particularly urinary tract infections. Both components of cotrimoxazole are synthetic antimicrobial drugs, and their combination was introduced into medical therapeutics about half a century ago. In Gram-negative bacteria, resistance to cotrimoxazole is widespread, being based on the acquisition of genes from the auxiliary genome that confer resistance to each of its antibacterial components. Starting from previous knowledge on the genotype of resistance to sulfamethoxazole in a collection of cotrimoxazole resistant uropathogenic Escherichia coli strains, this work focused on the identification of the genetic bases of the trimethoprim resistance of these same strains. Molecular techniques employed included PCR and Sanger sequencing of specific amplicons, conjugation experiments and NGS sequencing of the transferred plasmids. Mobile genetic elements conferring the trimethoprim resistance phenotype were identified and included integrons, transposons and single gene cassettes. Therefore, strains exhibited several ways to jointly resist both antibiotics, implying different levels of genetic linkage between genes conferring resistance to sulfamethoxazole (sul) and trimethoprim (dfrA). Two structures were particularly interesting because they represented a highly cohesive arrangements ensuring cotrimoxazole resistance. They both carried a single gene cassette, dfrA14 or dfrA1, integrated in two different points of a conserved cluster sul2-strA-strB, carried on transferable plasmids. The results suggest that the pressure exerted by cotrimoxazole on bacteria of our environment is still promoting the evolution toward increasingly compact gene arrangements, carried by mobile genetic elements that move them in the genome and also transfer them horizontally among bacteria.
RESUMO
Uropathogenic Escherichia coli is the leading etiologic agent of urinary tract infections, encompassing a highly heterogeneous group of strains. Although many putative urovirulence factors have been described, none of them appear in all uropathogenic E. coli strains, a fact that suggests that this group would be composed of different pathogenic subgroups. In this work, a study was performed on two collections of E. coli isolates proceeding from urine cultures from two groups of patients with urinary tract infection: pregnant women and children with urinary tract abnormalities. The isolates were analyzed for their virulence content and for their phylogeny by means of PCR determinations and of phenotypic assays. Associations among the virulence traits analyzed were searched for and this approach led to the identification of five urovirulence profiles. From a total of 230 isolates, 123 (53%) could be assigned to one of these profiles. A few loci appeared as markers of these profiles so that their presence allowed predicting the general virulence content of the strains. It is presumed that these conserved associations among the virulence functions would be devoted to ensure the coherence of the bacterial pathogenic strategy. In addition, three profiles appeared with significantly different frequencies depending on the host of origin of the isolates, indicating the existence of a correlation between the virulence content of the strains and their host specificity.
Assuntos
Infecções por Escherichia coli/microbiologia , Infecções Urinárias/microbiologia , Escherichia coli Uropatogênica/isolamento & purificação , Fatores de Virulência/genética , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Filogenia , Gravidez , Gestantes , Escherichia coli Uropatogênica/classificação , Escherichia coli Uropatogênica/genética , Escherichia coli Uropatogênica/metabolismo , Fatores de Virulência/metabolismo , Adulto JovemRESUMO
Urinary tract infections are among the most common infectious diseases encountered in humans and Escherichia coli is their leading etiologic agent. Uropathogenic E. coli encompasses a group of bacteria possessing a variable virulence gene assortment. It is generally agreed that many urovirulence factors remain to be discovered and that this information is required to gain knowledge on the pathogenic processes underlying the different clinical presentations of urinary tract infections. The production of higher-molecular-mass microcins, a group of ribosomally-synthesized peptide antibiotics comprising microcins H47, I47, E492, M and ColV, has been proposed as a virulence trait of some uropathogenic E. coli. To study this possibility, clones producing any of these microcins were selected from a collection of 160 Gram-negative clinical isolates from urine cultures and their virulence profile was analyzed. The study consisted in surveying genetic loci known to be relevant to urinary tract infection caused by E. coli. Depending on the type of microcin produced, different virulence patterns were observed which seemed to be determined by the degree of compatibility between virulence and microcin loci. In conclusion, results pointed to a relationship between higher-molecular-mass microcins and urovirulence.
Assuntos
Bacteriocinas/biossíntese , Infecções por Escherichia coli/microbiologia , Infecções Urinárias/microbiologia , Escherichia coli Uropatogênica/genética , Escherichia coli Uropatogênica/patogenicidade , Fatores de Virulência/biossíntese , Bacteriocinas/genética , Genótipo , Humanos , Reação em Cadeia da Polimerase/métodos , Escherichia coli Uropatogênica/isolamento & purificação , Fatores de Virulência/genéticaRESUMO
Microcins are ribosomally synthesized peptide antibiotics that are produced by enterobacterial strains. Although the first studies concentrated on plasmid-encoded activities, in the last years three chromosome-encoded microcins have been described: H47, E492, and M. Here, a new microcin, I47, is presented as a fourth member of this group. Common features exhibited by chromosome-encoded microcins were searched for. The comparison of the genetic clusters responsible for microcin production revealed a preserved general scheme. The clusters essentially comprise a pair of activity-immunity genes which determine antibiotic specificity and a set of microcin maturation and secretion genes which are invariably present and whose protein products are highly homologous among the different producing strains. A strict functional relationship between the maturation and secretion pathways of microcins H47, I47, and E492 was demonstrated through genetic analyses, which included heterologous complementation assays. The peptide precursors of these microcins share a maturation process which implies the addition of a catecholate siderophore of the salmochelin type. Microcins thus acquire the ability to enter gram-negative cells through the catechol receptors. In addition, they employ a common mode of secretion to reach the external milieu by means of a type I export apparatus. The results presented herein lead us to propose that chromosome-encoded microcins constitute a defined subgroup of peptide antibiotics which are strictly related by their modes of synthesis, secretion, and uptake.