A culture medium for screening 16S rRNA methylase-producing pan-aminoglycoside resistant Gram-negative bacteria

Nordmann, Patrice; Mazé, Alain; Culebras, Esther; Dobiáš, J; Jayol, Aurélie; Poirel, Laurent

doi:10.1016/j.diagmicrobio.2018.01.026

Cited by 7 publications

(6 citation statements)

References 14 publications

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“…Rectal swabs were cultured two hours after collection in Luria–Bertani broth (LBB, Roth, Karlsruhe, Germany) overnight. After LBB enrichment, a 10 µL loop of each broth was struck onto four different selective media, namely ChromID ESBL (bioMérieux, La Balme-La-Grotte, France) to select extended-spectrum β-lactamase (ESBL)-producing Gram-negatives, ChromID Carba Smart (bioMérieux) to select carbapenem-resistant Gram-negatives, home-made SuperAminoglycoside supplemented with amikacin 30 mg/L and gentamicin 30 mg/L to select pan-aminoglycoside-resistant Gram-negatives, and SuperPolymyxin supplemented with colistin 3.5 mg/L (EliTech Microbio, Signes, France) to select colistin-resistant Gram-negatives [ 6 , 7 ]. Grown colonies were tested by oxidase test (bioMérieux), and positive results were excluded as nonfermenting Gram-negative bacteria.…”

Section: Methodsmentioning

confidence: 99%

Does an Antibiotic Stewardship Applied in a Pig Farm Lead to Low ESBL Prevalence?

Fournier

Nordmann

Pittet³

et al. 2021

Antibiotics

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Background. The aim of the present study was to prospectively evaluate the prevalence of intestinal carriage of colistin-resistant and extended-spectrum β-lactamase (ESBL)-producing Enterobacterales among pigs from a Swiss farm attending an animal health and antibiotic stewardship program and to determine the associated mechanisms of resistance. Materials/Methods. Eighty-one fecal samples were recovered and screened for either β-lactam-resistant, colistin-resistant, or aminoglycoside-resistant Enterobacterales, using respective screening media. All recovered isolates were tested for antimicrobial susceptibility and their clonal relationship (PFGE and MLST). Plasmid typing was performed by plasmid-based replicon typing (PBRT). Resistance genes were searched by PCR and sequencing. Results. A total of 38 ESBL-producing Escherichia coli and a single ESBL-producing Enterobacter cloacae were recovered from 81 pigs, corresponding to a prevalence of 50%, no other β-lactamase producer being identified. Among the 38 ESBL-producing E. coli, all belonged to sequence type (ST) ST10, except two ST34 and ST744 isolates. Among the ST10-blaCTX-M-1 isolates, three subclones (n = 22, n = 13, and n = 1, respectively) were identified according to the PFGE analysis. The most commonly identified IncI1 plasmid harboring the blaCTX-M-1 gene was 143 kb in size and coharbored other resistance genes. Only three colistin-resistant Enterobacterales isolates were recovered, namely two Klebsiella pneumoniae isolates and a single E. cloacae isolate. Screening for the plasmid-borne mcr-1 to mcr-9 genes in these three isolates gave negative results. The two K. pneumoniae isolates were clonally related, belonged to ST76, and harbored a truncated mgrB chromosomal gene being the source of colistin resistance. Conclusion. A high prevalence of fecal carriage of ESBL-producing E. coli was found, being mainly caused by the spread of a clonal lineage within the farm. By contrast, a low prevalence of colistin-resistant Enterobacterales was found.

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Section: Methodsmentioning

confidence: 99%

Does an Antibiotic Stewardship Applied in a Pig Farm Lead to Low ESBL Prevalence?

Fournier

Nordmann

Pittet³

et al. 2021

Antibiotics

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“…The samples were incubated in Luria-Bertani (LB) broth (5 ml) overnight. From each sample, one calibrated loop (10 l) was plated on each of the three selective media, i.e., (i) CHROMagar mSuperCARBA (CHROMagar, Paris, France) (23), (ii) SuperAminoglycoside (Drigalski agar supplemented with 30 g/ml gentamicin, 30 g/ml amikacin, 10 g/ml vancomycin, and 5 g/ml amphotericin B) (24), and (iii) SuperPolymyxin (eosin methylene blue agar supplemented with 1 g/ml colistin, 10 g/ml daptomycin, and 5 g/ml amphotericin B) (25). The isolates that were selected were identified at the species level using the API20E system (bioMérieux, La Balme-les-Grottes, France).…”

Section: Methodsmentioning

confidence: 99%

Screening and Characterization of Multidrug-Resistant Gram-Negative Bacteria from a Remote African Area, São Tomé and Príncipe

Poirel

Aires-de-Sousa

Kudyba

et al. 2018

Antimicrob Agents Chemother

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The occurrence of resistance to last-resort antibiotics was evaluated among isolates recovered from hospitalized children in a remote African archipelago, São Tomé and Príncipe, where there is limited access to those antibiotics. Fifty patients were screened for colonization by carbapenem-, pan-aminoglycoside-, or polymyxin-resistant A total of 36 isolates (including 30 and 4) were recovered from 23 patients, including 26 isolates harboring the carbapenemase gene, a single isolate harboring the 16S rRNA methylase gene encoding pan-resistance to aminoglycosides, and 8 isolates coharboring both genes. A single isolate possessed the plasmid-borne colistin resistance gene A high clonal relationship was found for OXA-181-producing (4 clones), and conversely, three of the four OXA-181-producing isolates were clonally unrelated. This study overall showed a high prevalence of resistance to last-resort antibiotics in this country, where no epidemiological data were previously available.

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“…One calibrated inoculated loop (10 μL) of each sample was plated on to SuperPolymyxin selective agar medium that contains 3.5 μg/mL of colistin and 10 μg/mL of daptomycin (ELITechGroup, Signes, France) to select for colistin-resistant Gram-negative bacteria [9] . In addition, screening of ESBL-producing isolates was performed using ChromID ESBL (bioMérieux, La Balme les Grottes, France) selective agar medium, screening of pan-aminoglycoside-resistant Gram-negative bacteria was performed using SuperAminoglycoside selective medium [10] , and screening of carbapenemase-producing Enterobacterales was performed using SuperCarba and CarbaSmart selective plates (bioMérieux) [11] . Antibiotic susceptibility testing was performed by disk diffusion assay, except for colistin which was tested by broth microdilution.…”

Section: Screening Of Resistant Isolatesmentioning

confidence: 99%

Occurrence of CTX-M-15- and MCR-1-producing Enterobacterales in pigs in Portugal: Evidence of direct links with antibiotic selective pressure

Fournier

Aires-de-Sousa

Nordmann

et al. 2020

International Journal of Antimicrobial Agents

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To undertake a prospective analysis of the occurrence of colistin-resistant and extended-spectrum ß-lactamase (ESBL)-producing Enterobacterales colonizing pigs at two farms in Portugal, and to evaluate the putative correlations with usage of different antibiotics. Materials and methods: One hundred and two faecal samples recovered from two different Portuguese pig farms were screened for polymyxin-resistant and ESBL-positive Enterobacterales. The authors had undertaken a study at one of the farms previously, but the use of colistin has since been banned; zinc oxide and amoxicillin are used as prophylactic and curative drugs, respectively, at this farm. The other farm included in this study used zinc oxide alone. Results: Ninety-three ESBL-producing isolates (62 Escherichia coli , 29 Klebsiella pneumoniae , one Enterobacter aerogenes and one Enterobacter cloacae ) and 17 colistin-resistant isolates (12 E. coli , four K. pneumoniae and one E. cloacae ) were recovered. Among the ESBL producers, the majority (84%) produced CTX-M-15, while the others produced CTX-M-1 or CTX-M-9. Many different strain and plasmid backgrounds were identified, ruling out a massive dissemination of one major clone. In total, 17 colistin-resistant isolates were recovered, all from the first farm. All produced MCR-1, corresponding to 12 E. coli (10 clones) and three K. pneumoniae (two clones). The MCR-1 producers were all recovered from the farm where colistin had been used 2 years previously. Conclusion:This study showed a surprisingly high rate of CTX-M-15 producers at two Portuguese pig farms. A link was found between antibiotic selective pressure (ß-lactam or polymyxin) and the corresponding resistance rate.

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A culture medium for screening 16S rRNA methylase-producing pan-aminoglycoside resistant Gram-negative bacteria

Cited by 7 publications

References 14 publications

Does an Antibiotic Stewardship Applied in a Pig Farm Lead to Low ESBL Prevalence?

Does an Antibiotic Stewardship Applied in a Pig Farm Lead to Low ESBL Prevalence?

Screening and Characterization of Multidrug-Resistant Gram-Negative Bacteria from a Remote African Area, São Tomé and Príncipe

Occurrence of CTX-M-15- and MCR-1-producing Enterobacterales in pigs in Portugal: Evidence of direct links with antibiotic selective pressure

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