title: High prevalence of OXA-143 and alteration of outer membrane proteins in carbapenem-resistant Acinetobacter spp. isolates in Brazil.
abstract: Carbapenem resistance amongst Acinetobacter spp. has been increasing in the last decade. This study evaluated the outer membrane protein (OMP) profile and production of carbapenemases in 50 carbapenem-resistant Acinetobacter spp. isolates from bloodstream infections. Isolates were identified by API20NE. Minimum inhibitory concentrations (MICs) for carbapenems were determined by broth microdilution. Carbapenemases were studied by phenotypic tests, detection of their encoding gene by polymerase chain reaction (PCR) amplification, and imipenem hydrolysis. Nucleotide sequencing confirming the enzyme gene type was performed using MegaBACE 1000. The presence of OMPs was studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and PCR. Molecular typing was performed using pulsed-field gel electrophoresis (PFGE). All isolates were resistant to carbapenems. Moreover, 98% of the isolates were positive for the gene encoding the enzyme OXA-51-like, 18% were positive for OXA-23-like (only one isolate did not show the presence of the insertion sequence ISAba1 adjacent to this gene) and 76% were positive for OXA-143 enzyme. Five isolates (10%) showed the presence of the IMP-1 gene. Imipenem hydrolysing activity was detected in only three strains containing carbapenemase genes, comprising two isolates containing the bla(IMP) gene and one containing the bla(OXA-51/OXA-23-like) gene. The OMP of 43 kDa was altered in 17 of 25 strains studied, and this alteration was associated with a high meropenem MIC (256 μg/mL) in 5 of 7 strains without 43 kDa OMP. On the other hand, decreased OMP 33-36 kDa was found in five strains. The high prevalence of OXA-143 and alteration of OMPs might have been associated with a high level of carbapenem resistance.
authors.affiliation: Laboratory of Bacteriology (LIM-54), Hospital das Clínicas, Medical School of the University of São Paulo, São Paulo, Brazil.
authors.forename: Anna Karina
authors.initials: AK
authors.lastname: Mostachio
authors.forename: Anna Sara
authors.initials: AS
authors.lastname: Levin
authors.forename: Camila
authors.initials: C
authors.lastname: Rizek
authors.forename: Flavia
authors.initials: F
authors.lastname: Rossi
authors.forename: Jessika
authors.initials: J
authors.lastname: Zerbini
authors.forename: Silvia Figueiredo
authors.initials: SF
authors.lastname: Costa
mesh_terms: D000150:Acinetobacter / Q000737:chemistry* / Q000187:drug effects / Q000201:enzymology* / Q000302:isolation & purification; D000151:Acinetobacter Infections / Q000453:epidemiology* / Q000382:microbiology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D016470:Bacteremia / Q000453:epidemiology / Q000382:microbiology; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D001938:Brazil / Q000453:epidemiology; D015780:Carbapenems / Q000494:pharmacology*; D016000:Cluster Analysis; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D058889:Molecular Typing; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D015995:Prevalence; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D015780:Carbapenems; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D001618:beta-Lactamases