Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections
Sixty-six clinical P. aeruginosa isolates, 17 obtained from canine otitis specimens and 49 received from human patients with bloodstream infections, were collected between February 2007 and January 2008. The minimal inhibitory concentrations (MICs) of the antimicrobial agents of these isolates were...
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| Format: | Article |
| Language: | English |
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Czech Academy of Agricultural Sciences
2010-04-01
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| Series: | Veterinární Medicína |
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| Online Access: | https://vetmed.agriculturejournals.cz/artkey/vet-201004-0004_molecular-mechanisms-of-ceftazidime-resistance-in-pseudomonas-aeruginosa-isolates-from-canine-and-human-infecti.php |
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| author | S.-J. Du H.-C. Kuo C.-H. Cheng A.C.Y. Fei H.-W. Wei S.-K. Chang |
| author_facet | S.-J. Du H.-C. Kuo C.-H. Cheng A.C.Y. Fei H.-W. Wei S.-K. Chang |
| author_sort | S.-J. Du |
| collection | DOAJ |
| description | Sixty-six clinical P. aeruginosa isolates, 17 obtained from canine otitis specimens and 49 received from human patients with bloodstream infections, were collected between February 2007 and January 2008. The minimal inhibitory concentrations (MICs) of the antimicrobial agents of these isolates were determined. Multidrug resistance was common, with 23 (34.8%) isolates found to be ceftazidime resistant. To explore the mechanisms of ceftazidime resistance, PCR analyses were performed to detect drug-resistance genes. The prevalence rate of Ambler class A, B, and D β-lactamase genes were obtained, with blaTEM-1 100%, blaPSE-1 100%, blaOXA-2 96.2%, blaSHV-18 91.3%, blaOXA-17 78.3%, blaVIM-3 26.1%, blaOXA-10 21.7% and blaSHV-1 8.7%. An efflux inhibition assay with the PAβN compound was conducted. The ceftazidime resistance isolates were also tested by RT-qPCR to determine the mRNA expression levels of the oprM and ampC genes. Five (21.7%) of the ceftazidime resistance isolates appeared to overactivate the OprM efflux system. The ampD, ampE, and ampR genes and the ampC-ampR intergenic region were subsequently amplified and sequenced. Five (21.7%) of the ceftazidime resistance isolates from humans and canines had a point mutation in AmpR (Asp135-Asn, n = 3; Als194-Ser, n = 2), which induces AmpC overproduction from 10- to 80-fold. This study first reported ceftazidime resistance in P. aeruginosa from canine otitis specimens, which are closely related to ESBLs (50%), including the overproduction of AmpC (25%) and the OprM efflux system (25%). The ESBLs (100%) played an important role in all ceftazidime resistance isolates from humans, and either AmpC (21.1%) or OprM (21.1%) might be overexpressed within the same isolate. A human patient isolate (H307B) showed simultaneous expression of ESBLs, the OprM efflux system, and AmpC overproduction. |
| first_indexed | 2024-04-10T08:01:00Z |
| format | Article |
| id | doaj.art-9ba8472885ac4234a18d44168c46a1ee |
| institution | Directory Open Access Journal |
| issn | 0375-8427 1805-9392 |
| language | English |
| last_indexed | 2024-04-10T08:01:00Z |
| publishDate | 2010-04-01 |
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| series | Veterinární Medicína |
| spelling | doaj.art-9ba8472885ac4234a18d44168c46a1ee2023-02-23T03:49:48ZengCzech Academy of Agricultural SciencesVeterinární Medicína0375-84271805-93922010-04-0155417218210.17221/64/2010-VETMEDvet-201004-0004Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infectionsS.-J. Du0H.-C. Kuo1C.-H. Cheng2A.C.Y. Fei3H.-W. Wei4S.-K. Chang5Graduate Institute of Veterinary Medicine, National Taiwan University, Taiwan, ROCDepartment of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan, ROCGraduate Institute of Veterinary Medicine, National Taiwan University, Taiwan, ROCGraduate Institute of Veterinary Medicine, National Taiwan University, Taiwan, ROCDepartment of Animal Science and Technology, National Taiwan University, Taipei, Taiwan, ROCGraduate Institute of Veterinary Medicine, National Taiwan University, Taiwan, ROCSixty-six clinical P. aeruginosa isolates, 17 obtained from canine otitis specimens and 49 received from human patients with bloodstream infections, were collected between February 2007 and January 2008. The minimal inhibitory concentrations (MICs) of the antimicrobial agents of these isolates were determined. Multidrug resistance was common, with 23 (34.8%) isolates found to be ceftazidime resistant. To explore the mechanisms of ceftazidime resistance, PCR analyses were performed to detect drug-resistance genes. The prevalence rate of Ambler class A, B, and D β-lactamase genes were obtained, with blaTEM-1 100%, blaPSE-1 100%, blaOXA-2 96.2%, blaSHV-18 91.3%, blaOXA-17 78.3%, blaVIM-3 26.1%, blaOXA-10 21.7% and blaSHV-1 8.7%. An efflux inhibition assay with the PAβN compound was conducted. The ceftazidime resistance isolates were also tested by RT-qPCR to determine the mRNA expression levels of the oprM and ampC genes. Five (21.7%) of the ceftazidime resistance isolates appeared to overactivate the OprM efflux system. The ampD, ampE, and ampR genes and the ampC-ampR intergenic region were subsequently amplified and sequenced. Five (21.7%) of the ceftazidime resistance isolates from humans and canines had a point mutation in AmpR (Asp135-Asn, n = 3; Als194-Ser, n = 2), which induces AmpC overproduction from 10- to 80-fold. This study first reported ceftazidime resistance in P. aeruginosa from canine otitis specimens, which are closely related to ESBLs (50%), including the overproduction of AmpC (25%) and the OprM efflux system (25%). The ESBLs (100%) played an important role in all ceftazidime resistance isolates from humans, and either AmpC (21.1%) or OprM (21.1%) might be overexpressed within the same isolate. A human patient isolate (H307B) showed simultaneous expression of ESBLs, the OprM efflux system, and AmpC overproduction.https://vetmed.agriculturejournals.cz/artkey/vet-201004-0004_molecular-mechanisms-of-ceftazidime-resistance-in-pseudomonas-aeruginosa-isolates-from-canine-and-human-infecti.phpampcamprextended spectrum β-lactamasesoprm efflux system |
| spellingShingle | S.-J. Du H.-C. Kuo C.-H. Cheng A.C.Y. Fei H.-W. Wei S.-K. Chang Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections Veterinární Medicína ampc ampr extended spectrum β -lactamases oprm efflux system |
| title | Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections |
| title_full | Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections |
| title_fullStr | Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections |
| title_full_unstemmed | Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections |
| title_short | Molecular mechanisms of ceftazidime resistance in Pseudomonas aeruginosa isolates from canine and human infections |
| title_sort | molecular mechanisms of ceftazidime resistance in pseudomonas aeruginosa isolates from canine and human infections |
| topic | ampc ampr extended spectrum β -lactamases oprm efflux system |
| url | https://vetmed.agriculturejournals.cz/artkey/vet-201004-0004_molecular-mechanisms-of-ceftazidime-resistance-in-pseudomonas-aeruginosa-isolates-from-canine-and-human-infecti.php |
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