The marine environment as a reservoir of enterococci carrying resistance and virulence genes strongly associated with clinical strains

To gain insights into the relationships and the genetic exchange among environmental and clinical enterococci, 59 strains (29 from marine aquaculture sites and 30 from clinical settings) resistant to tetracycline, erythromycin, ampicillin and/or gentamicin were analysed for the antibiotic resistance tet(M), tet(L), tet(O), erm(A), erm(B), mef blaZ, aac(6')-Ie aph(2″)-Ia and virulence gelE, cylB, efaA and esp genes, and for the copper resistance gene tcrB. Antibiotic resistance and virulence genes were detected more frequently in clinical than in environmental enterococci; the opposite was true for copper resistance. Conjugation experiments demonstrated the transfer of antibiotic resistance genes from marine to clinical enterococci in interspecific mating and the uncommon joint transfer of tet(L) and erm(B). Enterobacterial repetitive intergenic consensus polymerase chain reaction typing evidenced a cluster (90% similarity) encompassing strains carrying multiple antibiotic resistance genes from both sets; the others marine isolates exhibited polyclonality and bore tcrB. Our results demonstrate that antibiotic-resistant marine enterococci bear antibiotic resistance genes transferable to humans and suggest that copper resistance, not observed among clinical strains, may be useful for survival in the environment, whereas virulence genes likely confer no advantage to enterococcal populations adapted to a lifestyle outside the host

Epidemic Escherichia coli ST131 and Enterococcus faecium ST17 in coastal marine sediments from an Italian beach

Fecal indicator bacteria (FIB) are used worldwide to assess water quality in coastal environments, but little is known about their genetic diversity and pathogenicity. This study examines the prevalence, antimicrobial resistance, virulence, and genetic diversity of FIB isolated from marine sediments from a central Adriatic seaside resort. FIB, recovered from 6 out of 7 sites, were significantly more abundant at sampling stations 300 m offshore than close to the shore. Escherichia coli accounted for 34.5% of fecal coliforms, and Enterococcus faecalis accounted for 32% of enterococci. Most isolates (27% of E. coli and 22% of enterococci) were recovered from the sediments that had the highest organic content. Multidrug-resistant E. coli (31%) and enterococci (22%) were found at nearly all sites, whereas 34.5% of E. coli and 28% of enterococci harboring multiple virulence factors were recovered from just two sites. Pulsed-field gel electrophoresis typing showed wide genetic diversity among isolates. Human epidemic clones ( E. coli ST131 and Enterococcus faecium ST17) were identified for the first time by multilocus sequence typing in an area where bathing had not been prohibited. These clones were from sites far removed from riverine inputs, suggesting a wide diffusion of pathogenic FIB in the coastal environment and a high public health risk

The marine environment as a reservoir of enterococci carrying resistance and virulence genes strongly associated with clinical strains

To gain insights into the relationships and the genetic exchange among environmental and clinical enterococci, 59 strains (29 from marine aquaculture sites and 30 from clinical settings) resistant to tetracycline, erythromycin, ampicillin and/or gentamicin were analysed for the antibiotic resistance tet(M), tet(L), tet(O), erm(A), erm(B), mef blaZ, aac(6\ue2\u80\ub2)-Ie aph(2\ue2\u80\ub3)-Ia and virulence gelE, cylB, efaA and esp genes, and for the copper resistance gene tcrB. Antibiotic resistance and virulence genes were detected more frequently in clinical than in environmental enterococci; the opposite was true for copper resistance. Conjugation experiments demonstrated the transfer of antibiotic resistance genes from marine to clinical enterococci in interspecific mating and the uncommon joint transfer of tet(L) and erm(B). Enterobacterial repetitive intergenic consensus polymerase chain reaction typing evidenced a cluster (90% similarity) encompassing strains carrying multiple antibiotic resistance genes from both sets; the others marine isolates exhibited polyclonality and bore tcrB. Our results demonstrate that antibiotic-resistant marine enterococci bear antibiotic resistance genes transferable to humans and suggest that copper resistance, not observed among clinical strains, may be useful for survival in the environment, whereas virulence genes likely confer no advantage to enterococcal populations adapted to a lifestyle outside the host. \uc2\ua9 2013 Society for Applied Microbiology and John Wiley & Sons Ltd

Aquaculture Can Promote the Presence and Spread of Antibiotic-Resistant Enterococci in Marine Sediments

Aquaculture is an expanding activity worldwide. However its rapid growth can affect the aquatic environment through release of large amounts of chemicals, including antibiotics. Moreover, the presence of organic matter and bacteria of different origin can favor gene transfer and recombination. Whereas the consequences of such activities on environmental microbiota are well explored, little is known of their effects on allochthonous and potentially pathogenic bacteria, such as enterococci. Sediments from three sampling stations (two inside and one outside) collected in a fish farm in the Adriatic Sea were examined for enterococcal abundance and antibiotic resistance traits using the membrane filter technique and an improved quantitative PCR. Strains were tested for susceptibility to tetracycline, erythromycin, ampicillin and gentamicin; samples were directly screened for selected tetracycline [tet(M), tet(L), tet(O)] and macrolide [erm(A), erm(B) and mef] resistance genes by newly-developed multiplex PCRs. The abundance of benthic enterococci was higher inside than outside the farm. All isolates were susceptible to the four antimicrobials tested, although direct PCR evidenced tet(M) and tet(L) in sediment samples from all stations. Direct multiplex PCR of sediment samples cultured in rich broth supplemented with antibiotic (tetracycline, erythromycin, ampicillin or gentamicin) highlighted changes in resistance gene profiles, with amplification of previously undetected tet(O), erm(B) and mef genes and an increase in benthic enterococcal abundance after incubation in the presence of ampicillin and gentamicin. Despite being limited to a single farm, these data indicate that aquaculture may influence the abundance and spread of benthic enterococci and that farm sediments can be reservoirs of dormant antibiotic-resistant bacteria, including enterococci, which can rapidly revive in presence of new inputs of organic matter. This reservoir may constitute an underestimated health risk and deserves further investigation. \uc2\ua9 2013 Di Cesare et al

Location of the fish farm and of the sampling stations.

<p>The map is from <a href="http://earthobservatory.nasa.gov/" target="_blank">http://earthobservatory.nasa.gov/</a>, image courtesy Jesse Allen.</p

Resistance genes detected before and after sediment incubation in antibiotic-supplemented BHI broth.

*<p>detected both in sediment and in broth.</p

Control strains used in PCR assays.

a<p>American Type Culture Collection.</p

Abundance of benthic enterococci.

<p>Enterococcal abundance in the farm sediments and at the control site was determined by qPCR before and after incubation with antibiotic-supplemented BHI broth. A, St. 1; B, St. 2 and C, St. 3. *Not detectable i.e.</p

Primer pairs used to detect resistance genes in PCR assays.

<p>Primer pairs used to detect resistance genes in PCR assays.</p