OBJECTIVES: The aac(6')-Ih gene encoding aminoglycoside 6'-N-acetyltransferase type I subtype h [AAC(6')-Ih] is plasmid-borne in Acinetobacter baumannii where it confers high-level amikacin resistance, but its origin remains unknown. We searched for the gene in the genomes of a collection of 133 Acinetobacter spp. and studied its species specificity, expression and dissemination. METHODS: Gene copy number was determined by quantitative PCR, expression by quantitative RT-PCR, MIC by microdilution and transfer by plasmid mobilization. RESULTS: The aac(6')-Ih gene was present in the chromosome of the two Acinetobacter gyllenbergii of the collection and was detected in all seven A. gyllenbergii clinical isolates. They had indistinguishable flanking regions indicating that the gene was intrinsic to this species. A. baumannii PIS Aba23 promoters were provided by insertion of ISAba23, which disrupted the Pnative promoter in A. gyllenbergii. Both types of promoters were similarly potent in Escherichia coli and A. baumannii. Aminoglycoside MICs for A. baumannii harbouring pIP1858 were higher than for A. gyllenbergii due to gene dosage. The non-self-transferable plasmid could be mobilized to other A. baumannii cells by the broad host range plasmid RP4. CONCLUSIONS: We have found the origin of aac(6')-Ih in A. gyllenbergii, a species isolated, although rarely, in humans, and documented that dissemination of this gene is restricted to the Acinetobacter genus.
- MeSH
- acetyltransferasy genetika metabolismus MeSH
- Acinetobacter baumannii klasifikace účinky léků enzymologie genetika MeSH
- aminoglykosidy metabolismus farmakologie MeSH
- antibakteriální látky metabolismus farmakologie MeSH
- bakteriální léková rezistence * MeSH
- genová dávka MeSH
- infekce bakteriemi rodu Acinetobacter mikrobiologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- plazmidy analýza MeSH
- přenos genů horizontální MeSH
- promotorové oblasti (genetika) MeSH
- transpozibilní elementy DNA MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
A new separation and quantification method using ultra high-performance liquid chromatography (UHPLC) with UV detection was developed for the detection of sibiromycin in fermentation broth of Streptosporangium sibiricum. The solid phase extraction method based on cation-exchange was employed to pre-concentrate and purify fermentation broth containing sibiromycin prior to UHPLC analysis. The whole assay was validated and showed a linear range of detector response for the quantification of sibiromycin in a concentration from 3.9 to 250.0 μg mL⁻¹, with correlation coefficient of 0.999 and recoveries ranging from 71.66±3.55% to 74.76±5.18%. Method limit of quantification of the assay was determined as 0.18 μg mL⁻¹ and was verified with resulting RSD of 9.6% and accuracy of 97.6%. The developed assay was used to determine the sibiromycin production in 12 different fermentation broths. Moreover, several natural sibiromycin analogues/derivatives were described with pilot characterization using off-line mass spectrometry: the previously described dihydro-sibiromycin (DH-sibiromycin) and tentative bis-glycosyl forms of sibiromycin and its dihydro-analogue.
- MeSH
- Actinomycetales metabolismus MeSH
- aminoglykosidy analýza chemie metabolismus MeSH
- extrakce na pevné fázi MeSH
- fermentace MeSH
- hmotnostní spektrometrie metody MeSH
- kalibrace MeSH
- kultivační média speciální chemie MeSH
- lineární modely MeSH
- molekulární struktura MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- stabilita léku MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- přírodní látky, antibiotika,
- MeSH
- acetyltransferasy metabolismus terapeutické užití MeSH
- aminoglykosidy metabolismus terapeutické užití MeSH
- aminokyseliny chemie klasifikace metabolismus MeSH
- antibakteriální látky biosyntéza metabolismus terapeutické užití MeSH
- arginin analogy a deriváty metabolismus MeSH
- bakteriální proteiny izolace a purifikace metabolismus terapeutické užití MeSH
- lidé MeSH
- lysin analogy a deriváty metabolismus MeSH
- nukleosidy izolace a purifikace metabolismus terapeutické užití MeSH
- streptothriciny metabolismus terapeutické užití MeSH
- Check Tag
- lidé MeSH
- MeSH
- aminoglykosidy aplikace a dávkování farmakokinetika metabolismus MeSH
- chinoliny aplikace a dávkování farmakokinetika metabolismus MeSH
- karbapenemy aplikace a dávkování farmakokinetika metabolismus MeSH
- kritický stav MeSH
- lidé MeSH
- piperacilin aplikace a dávkování farmakokinetika metabolismus MeSH
- renální insuficience farmakoterapie MeSH
- rozvrh dávkování léků MeSH
- Check Tag
- lidé MeSH
An in vitro evaluation of 32 gentamicin-resistant, Gram-negative bacteria from hygiene-centres in Berne, Switzerland and Bratislava, Czechoslovakia, revealed that 24 strains produced gentamicin-modifying enzymes. Occurrence of acetyltransferases (AAC) was confirmed in 16 strains, adenylyltransferase (AAD) /2''/ was observed in 13 strains while 5 of the gentamicin-resistant strains produced both enzymes. All Czechoslovak strains were amikacin-susceptible, in amikacin-resistant Pseudomonas aeruginosa strains from Switzerland the presence of AAC/6'/ was found. In the majority of gentamicin-resistant, netilmicin-susceptible strains the occurrence of AAD/2''/ was observed. Gentamicin-resistance in the modifying enzymes producing strains was due either to production of an acetyltransferase or the adenylyltransferase except Swiss strains of Serratia marcescens where a simultaneous production of both types of the enzymes was noted. Twenty eight strains produced an enzyme modifying aminoglycosides of any kind.
- MeSH
- acetyltransferasy biosyntéza MeSH
- aminoglykosidy metabolismus farmakologie MeSH
- antibakteriální látky metabolismus farmakologie MeSH
- antibiotická rezistence MeSH
- bakteriální infekce mikrobiologie MeSH
- druhová specificita MeSH
- fosfotransferasy biosyntéza MeSH
- gentamiciny farmakologie MeSH
- gramnegativní bakterie účinky léků enzymologie metabolismus MeSH
- kanamycinkinasa MeSH
- lidé MeSH
- nukleotidyltransferasy biosyntéza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Československo MeSH
- Švýcarsko MeSH
- MeSH
- acetyltransferasy metabolismus MeSH
- aminoglykosidy metabolismus farmakologie MeSH
- antibakteriální látky metabolismus farmakologie MeSH
- antibiotická rezistence MeSH
- Enterobacteriaceae účinky léků enzymologie MeSH
- fosfotransferasy metabolismus MeSH
- nukleotidyltransferasy metabolismus MeSH
- Pseudomonas aeruginosa účinky léků enzymologie MeSH
- Pseudomonas enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Geografické názvy
- Československo MeSH
- Německo západní MeSH
- Švýcarsko MeSH
