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Dogs leaving the ICU carry a very large multi-drug resistant enterococcal population with capacity for biofilm formation and horizontal gene transfer.

Ghosh A, Dowd SE, Zurek L - PLoS ONE (2011)

Bottom Line: In vitro intra-species conjugation assays demonstrated that E. faecium were capable of transferring tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin resistance traits to human clinical strains.Multi-locus variable number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE) of E. faecium strains showed very low genotypic diversity.It is recommended to restrict close physical contact between pets released from the ICU and their owners to avoid potential health risks.

View Article: PubMed Central - PubMed

Affiliation: Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, United States of America.

ABSTRACT
The enterococcal community from feces of seven dogs treated with antibiotics for 2-9 days in the veterinary intensive care unit (ICU) was characterized. Both, culture-based approach and culture-independent 16S rDNA amplicon 454 pyrosequencing, revealed an abnormally large enterococcal community: 1.4±0.8×10(8) CFU gram(-1) of feces and 48.9±11.5% of the total 16,228 sequences, respectively. The diversity of the overall microbial community was very low which likely reflects a high selective antibiotic pressure. The enterococcal diversity based on 210 isolates was also low as represented by Enterococcus faecium (54.6%) and Enterococcus faecalis (45.4%). E. faecium was frequently resistant to enrofloxacin (97.3%), ampicillin (96.5%), tetracycline (84.1%), doxycycline (60.2%), erythromycin (53.1%), gentamicin (48.7%), streptomycin (42.5%), and nitrofurantoin (26.5%). In E. faecalis, resistance was common to tetracycline (59.6%), erythromycin (56.4%), doxycycline (53.2%), and enrofloxacin (31.9%). No resistance was detected to vancomycin, tigecycline, linezolid, and quinupristin/dalfopristin in either species. Many isolates carried virulence traits including gelatinase, aggregation substance, cytolysin, and enterococcal surface protein. All E. faecalis strains were biofilm formers in vitro and this phenotype correlated with the presence of gelE and/or esp. In vitro intra-species conjugation assays demonstrated that E. faecium were capable of transferring tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin resistance traits to human clinical strains. Multi-locus variable number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE) of E. faecium strains showed very low genotypic diversity. Interestingly, three E. faecium clones were shared among four dogs suggesting their nosocomial origin. Furthermore, multi-locus sequence typing (MLST) of nine representative MLVA types revealed that six sequence types (STs) originating from five dogs were identical or closely related to STs of human clinical isolates and isolates from hospital outbreaks. It is recommended to restrict close physical contact between pets released from the ICU and their owners to avoid potential health risks.

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Related in: MedlinePlus

Biofilm formation, gelatinase (protease) phenotype and presence of esp in enterococci from the feces of ICU dogs.(A) E. faecalis (n = 90), (B) E. faecium (n = 85). The dashed lines indicate biofilm formation activity (<0.2 = no biofilm, 0.2–0.7 = biofilm, >0.7 = strong biofilm). Bars correspond to the mean ± SEM of 5 replicates. E. faecalis V583 used as a positive control.
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pone-0022451-g004: Biofilm formation, gelatinase (protease) phenotype and presence of esp in enterococci from the feces of ICU dogs.(A) E. faecalis (n = 90), (B) E. faecium (n = 85). The dashed lines indicate biofilm formation activity (<0.2 = no biofilm, 0.2–0.7 = biofilm, >0.7 = strong biofilm). Bars correspond to the mean ± SEM of 5 replicates. E. faecalis V583 used as a positive control.

Mentions: All E. faecalis isolates tested were biofilm formers (OD595>0.2) and many of them (47/90, 52.2%) produced a strong biofilm (OD595>0.7) (Figure 4A). Overall, biofilm formation correlated with the presence of strong gelatinase phenotype and/or with the esp gene. In contrast, none of E. faecium formed biofilm and all of them lacked the strong gelatinase phenotype as well as esp (Figure 4B).


Dogs leaving the ICU carry a very large multi-drug resistant enterococcal population with capacity for biofilm formation and horizontal gene transfer.

Ghosh A, Dowd SE, Zurek L - PLoS ONE (2011)

Biofilm formation, gelatinase (protease) phenotype and presence of esp in enterococci from the feces of ICU dogs.(A) E. faecalis (n = 90), (B) E. faecium (n = 85). The dashed lines indicate biofilm formation activity (<0.2 = no biofilm, 0.2–0.7 = biofilm, >0.7 = strong biofilm). Bars correspond to the mean ± SEM of 5 replicates. E. faecalis V583 used as a positive control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3139645&req=5

pone-0022451-g004: Biofilm formation, gelatinase (protease) phenotype and presence of esp in enterococci from the feces of ICU dogs.(A) E. faecalis (n = 90), (B) E. faecium (n = 85). The dashed lines indicate biofilm formation activity (<0.2 = no biofilm, 0.2–0.7 = biofilm, >0.7 = strong biofilm). Bars correspond to the mean ± SEM of 5 replicates. E. faecalis V583 used as a positive control.
Mentions: All E. faecalis isolates tested were biofilm formers (OD595>0.2) and many of them (47/90, 52.2%) produced a strong biofilm (OD595>0.7) (Figure 4A). Overall, biofilm formation correlated with the presence of strong gelatinase phenotype and/or with the esp gene. In contrast, none of E. faecium formed biofilm and all of them lacked the strong gelatinase phenotype as well as esp (Figure 4B).

Bottom Line: In vitro intra-species conjugation assays demonstrated that E. faecium were capable of transferring tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin resistance traits to human clinical strains.Multi-locus variable number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE) of E. faecium strains showed very low genotypic diversity.It is recommended to restrict close physical contact between pets released from the ICU and their owners to avoid potential health risks.

View Article: PubMed Central - PubMed

Affiliation: Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, United States of America.

ABSTRACT
The enterococcal community from feces of seven dogs treated with antibiotics for 2-9 days in the veterinary intensive care unit (ICU) was characterized. Both, culture-based approach and culture-independent 16S rDNA amplicon 454 pyrosequencing, revealed an abnormally large enterococcal community: 1.4±0.8×10(8) CFU gram(-1) of feces and 48.9±11.5% of the total 16,228 sequences, respectively. The diversity of the overall microbial community was very low which likely reflects a high selective antibiotic pressure. The enterococcal diversity based on 210 isolates was also low as represented by Enterococcus faecium (54.6%) and Enterococcus faecalis (45.4%). E. faecium was frequently resistant to enrofloxacin (97.3%), ampicillin (96.5%), tetracycline (84.1%), doxycycline (60.2%), erythromycin (53.1%), gentamicin (48.7%), streptomycin (42.5%), and nitrofurantoin (26.5%). In E. faecalis, resistance was common to tetracycline (59.6%), erythromycin (56.4%), doxycycline (53.2%), and enrofloxacin (31.9%). No resistance was detected to vancomycin, tigecycline, linezolid, and quinupristin/dalfopristin in either species. Many isolates carried virulence traits including gelatinase, aggregation substance, cytolysin, and enterococcal surface protein. All E. faecalis strains were biofilm formers in vitro and this phenotype correlated with the presence of gelE and/or esp. In vitro intra-species conjugation assays demonstrated that E. faecium were capable of transferring tetracycline, doxycycline, streptomycin, gentamicin, and erythromycin resistance traits to human clinical strains. Multi-locus variable number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE) of E. faecium strains showed very low genotypic diversity. Interestingly, three E. faecium clones were shared among four dogs suggesting their nosocomial origin. Furthermore, multi-locus sequence typing (MLST) of nine representative MLVA types revealed that six sequence types (STs) originating from five dogs were identical or closely related to STs of human clinical isolates and isolates from hospital outbreaks. It is recommended to restrict close physical contact between pets released from the ICU and their owners to avoid potential health risks.

Show MeSH
Related in: MedlinePlus