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McCormack A, Hobbs JK, Johnston PR, Coote PJ. Surviving Colonies of Pseudomonas aeruginosa Isolated In Vivo from Infected, Antibiotic-Treated Galleria mellonella Larvae Acquire an Antibiotic-Tolerant Phenotype. Antibiotics (Basel) 2025; 14:507. [PMID: 40426573 DOI: 10.3390/antibiotics14050507] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/04/2025] [Revised: 05/09/2025] [Accepted: 05/12/2025] [Indexed: 05/29/2025] Open
Abstract
Background: The aim of this work was to induce the formation of antibiotic-tolerant and/or persister cells in vivo using antibiotic therapy on Galleria mellonella larvae infected with P. aeruginosa, isolate these surviving cells, and characterise their phenotype and genotype. Methods: Infected larvae were treated with effective doses of either ceftazidime or meropenem. Despite this, surviving P. aeruginosa colonies were isolated from living larvae, and antibiotic killing, fitness, virulence, antibiotic resistance and the whole genome sequence of a selection of these isolates were compared with their original parent strains. Results: The surviving isolates had an increased minimum duration to kill 99% of the population (MDK99) upon exposure to ceftazidime or meropenem and decreased growth rates in culture, but they showed no change to the MIC or virulence-consistent with an antibiotic-tolerant phenotype. Long-read genome sequencing of selected isolates revealed only one single nucleotide polymorphism (SNP) within bkdB, encoding the lipoamide acyltransferase component of the branched-chain α-keto acid dehydrogenase, present in two independent isolates. However, time-kill assays with ceftazidime of bkdB knockout strains showed no significant change in the MDK99. Concomitant with the antibiotic-tolerant phenotype, many of the isolates also had a reduced rate of killing when exposed to heat stress. Conclusions: P. aeruginosa cells that survived antibiotic therapy in vivo were found to be antibiotic-tolerant and thermotolerant but not antibiotic-resistant and had reduced growth rates under optimal conditions but unchanged virulence. In the absence of a convincing genetic explanation, the co-induction of enhanced thermotolerance with antibiotic tolerance indicated that both are conferred by a heritable phenotypic mechanism.
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Affiliation(s)
- Alex McCormack
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, Fife, UK
| | - Joanne K Hobbs
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, Fife, UK
| | - Paul R Johnston
- School of Medicine, University of St Andrews, The North Haugh, St Andrews KY16 9TF, Fife, UK
| | - Peter J Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, Fife, UK
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Svedholm E, Bruce B, Parcell BJ, Coote PJ. Repurposing Mitomycin C in Combination with Pentamidine or Gentamicin to Treat Infections with Multi-Drug-Resistant (MDR) Pseudomonas aeruginosa. Antibiotics (Basel) 2024; 13:177. [PMID: 38391563 PMCID: PMC10886254 DOI: 10.3390/antibiotics13020177] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2024] [Revised: 02/05/2024] [Accepted: 02/08/2024] [Indexed: 02/24/2024] Open
Abstract
The aims of this study were (i) to determine if the combination of mitomycin C with pentamidine or existing antibiotics resulted in enhanced efficacy versus infections with MDR P. aeruginosa in vivo; and (ii) to determine if the doses of mitomycin C and pentamidine in combination can be reduced to levels that are non-toxic in humans but still retain antibacterial activity. Resistant clinical isolates of P. aeruginosa, a mutant strain over-expressing the MexAB-OprM resistance nodulation division (RND) efflux pump and a strain with three RND pumps deleted, were used. MIC assays indicated that all strains were sensitive to mitomycin C, but deletion of three RND pumps resulted in hypersensitivity and over-expression of MexAB-OprM caused some resistance. These results imply that mitomycin C is a substrate of the RND efflux pumps. Mitomycin C monotherapy successfully treated infected Galleria mellonella larvae, albeit at doses too high for human administration. Checkerboard and time-kill assays showed that the combination of mitomycin C with pentamidine, or the antibiotic gentamicin, resulted in synergistic inhibition of most P. aeruginosa strains in vitro. In vivo, administration of a combination therapy of mitomycin C with pentamidine, or gentamicin, to G. mellonella larvae infected with P. aeruginosa resulted in enhanced efficacy compared with monotherapies for the majority of MDR clinical isolates. Notably, the therapeutic benefit conferred by the combination therapy occurred with doses of mitomycin C close to those used in human medicine. Thus, repurposing mitomycin C in combination therapies to target MDR P. aeruginosa infections merits further investigation.
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Affiliation(s)
- Elin Svedholm
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Benjamin Bruce
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Benjamin J Parcell
- NHS Tayside, Medical Microbiology, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK
| | - Peter J Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
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3
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Fletcher M, McCormack A, Parcell BJ, Coote PJ. Combination Therapy with Ciprofloxacin and Pentamidine against Multidrug-Resistant Pseudomonas aeruginosa: Assessment of In Vitro and In Vivo Efficacy and the Role of Resistance-Nodulation-Division (RND) Efflux Pumps. Antibiotics (Basel) 2023; 12:1236. [PMID: 37627656 PMCID: PMC10451767 DOI: 10.3390/antibiotics12081236] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2023] [Revised: 07/24/2023] [Accepted: 07/25/2023] [Indexed: 08/27/2023] Open
Abstract
The aim of this work was to (i) evaluate the efficacy of a combination treatment of pentamidine with ciprofloxacin against Galleria mellonella larvae infected with an MDR strain of P. aeruginosa and (ii) determine if pentamidine acts as an efflux-pump inhibitor. Resistant clinical isolates, mutant strains overexpressing one of three RND efflux pumps (MexAB-OprM, MexCD-OprJ, and MexEF-OprN), and a strain with the same three pumps deleted were used. MIC assays confirmed that the clinical isolates and the mutants overexpressing efflux pumps were resistant to ciprofloxacin and pentamidine. The deletion of the three efflux pumps induced sensitivity to both compounds. Exposure to pentamidine and ciprofloxacin in combination resulted in the synergistic inhibition of all resistant strains in vitro, but no synergy was observed versus the efflux-pump deletion strain. The treatment of infected G. mellonella larvae with the combination of pentamidine and ciprofloxacin resulted in enhanced efficacy compared with the monotherapies and significantly reduced the number of proliferating bacteria. Our measurement of efflux activity from cells revealed that pentamidine had a specific inhibitory effect on the MexCD-OprJ and MexEF-OprN efflux pumps. However, the efflux activity and membrane permeability assays revealed that pentamidine also disrupted the membrane of all cells. In conclusion, pentamidine does possess some efflux-pump inhibitory activity, in addition to a more general disruptive effect on membrane integrity that accounts for its ability to potentiate ciprofloxacin activity. Notably, the enhanced efficacy of combination therapy with pentamidine and ciprofloxacin versus MDR P. aeruginosa strains in vivo merits further investigation into its potential to treat infections via this pathogen in patients.
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Affiliation(s)
- Megan Fletcher
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK; (M.F.); (A.M.)
| | - Alex McCormack
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK; (M.F.); (A.M.)
| | - Benjamin J. Parcell
- NHS Tayside, Medical Microbiology, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK;
| | - Peter J. Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK; (M.F.); (A.M.)
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Mackay B, Parcell BJ, Shirran SL, Coote PJ. Carbapenem-Only Combination Therapy against Multi-Drug Resistant Pseudomonas aeruginosa: Assessment of In Vitro and In Vivo Efficacy and Mode of Action. Antibiotics (Basel) 2022; 11:1467. [PMID: 36358122 PMCID: PMC9686798 DOI: 10.3390/antibiotics11111467] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/06/2022] [Revised: 10/20/2022] [Accepted: 10/21/2022] [Indexed: 04/28/2024] Open
Abstract
The aim of the study was to determine the efficacy of carbapenem-only combination treatments derived from four approved drugs (meropenem, doripenem, ertapenem and imipenem) against a MDR strain of P. aeruginosa in a Galleria mellonella larvae infection model. G. mellonella larvae were infected with P. aeruginosa NCTC 13437 (carrying the VIM 10 carbapenamase) and the efficacy of the six possible dual, four triple, and one quadruple carbapenem combination(s) were compared to their constituent monotherapies. Four of these combinations showed significantly enhanced survival compared to monotherapies and reduced the bacterial burden inside infected larvae but without complete elimination. Bacteria that survived combination therapy were slower growing, less virulent but with unchanged carbapenem MICs-observations that are consistent with a persister phenotype. In vitro time-kill assays confirmed that the combinations were bactericidal and confirmed that a low number of bacteria survived exposure. Mass spectrometry was used to quantify changes in the concentration of carbapenems in the presence of carbapenemase-carrying P. aeruginosa. The rate of degradation of individual carbapenems was altered, and often significantly reduced, when the drugs were in combinations compared with the drugs alone. These differences may account for the enhanced inhibitory effects of the combinations against carbapenem-resistant P. aeruginosa and are consistent with a 'shielding' hypothesis. In conclusion, carbapenem combinations show promise in combating MDR P. aeruginosa and are worthy of additional study and development.
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Affiliation(s)
- Brendan Mackay
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Benjamin J. Parcell
- NHS Tayside, Medical Microbiology, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK
| | - Sally L. Shirran
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Peter J. Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
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Detection of OXA-181-producing Pseudomonas aeruginosa in Germany. Int J Med Microbiol 2022; 312:151557. [PMID: 35842996 DOI: 10.1016/j.ijmm.2022.151557] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/24/2022] [Revised: 06/08/2022] [Accepted: 07/06/2022] [Indexed: 11/23/2022] Open
Abstract
OBJECTIVES To report the detection of the class D carbapenemase OXA-181 in an MDR clinical Pseudomonas aeruginosa isolate in Germany. METHODS Carbapenemase detection was performed by using several phenotypic tests such as the modified Hodge test, a combined disc test with boronic acid, EDTA or cloxacillin, a lysate-based inhibition assays and by PCR for common and rare carbapenemase genes. Antibiotic susceptibilities were determined by broth microdilution. The genetic environment of blaOXA-181 in the clinical P. aeruginosa isolate was characterised by Illumina and MinION sequencing. RESULTS An multidrug-resistant P. aeruginosa was isolated from a tracheal swab in 2019 and was sent to the German National Reference Centre for multidrug-resistant Gram-negative bacteria for carbapenemase detection. Several phenotypic tests indicated the presence of a carbapenemase which was not inhibited by EDTA nor by boronic acid. PCRs for common and rare carbapenemase genes revealed the presence of a blaOXA-181 gene. WGS data confirmed that the gene was located on the chromosome as part of a Tn2013 transposon. The genetic organisation of blaOXA-181 has already been described in a P. aeruginosa isolate from England, but both isolates differed significantly in their sequence types (ST111/ST235). Analysis of the genetic environment of the blaOXA-181 gene also revealed high homology to a plasmid from a Klebsiella pneumoniae isolate. CONCLUSIONS To our knowledge, this is the first report of blaOXA-181 in a clinical P. aeruginosa isolate in Germany which emphasises the ongoing spread of yet unusual carbapenemases among different Gram-negative species and therefore complicating their detection in routine laboratories.
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Ndugire W, Raviranga NGH, Lao J, Ramström O, Yan M. Gold Nanoclusters as Nanoantibiotic Auranofin Analogues. Adv Healthc Mater 2022; 11:e2101032. [PMID: 34350709 PMCID: PMC8816973 DOI: 10.1002/adhm.202101032] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/26/2021] [Revised: 07/13/2021] [Indexed: 12/21/2022]
Abstract
Auranofin, a gold(I)-complex with tetraacetylated thioglucose (Ac4 GlcSH) and triethylphosphine ligands, is an FDA-approved drug used as an anti-inflammatory aid in the treatment of rheumatoid arthritis. In repurposing auranofin for other diseases, it was found that the drug showed significant activity against Gram-positive but was inactive against Gram-negative bacteria. Herein, the design and synthesis of gold nanoclusters (AuNCs) based on the structural motif of auranofin are reported. Phosphine-capped AuNCs are synthesized and glycosylated, yielding auranofin analogues with mixed triphenylphosphine monosulfonate (TPPMS)/Ac4 GlcSH ligand shells. These AuNCs are active against both Gram-negative and Gram-positive bacteria, including multidrug-resistant pathogens. Notably, an auranofin analogue, a mixed-ligand 1.6 nm AuNC 4b, is more active than auranofin against Pseudomonas aeruginosa, while exhibiting lower toxicity against human A549 cells. The enhanced antibacterial activity of these AuNCs is characterized by a greater uptake of Au by the bacteria compared to AuI complexes. Additional factors include increased oxidative stress, moderate inhibition of thioredoxin reductase (TrxR), and DNA damage. Most intriguingly, the uptake of AuNCs are not affected by the bacterial outer membrane (OM) barrier or by binding with the extracellular proteins. This contrasts with AuI complexes like auranofin that are susceptible to protein binding and hindered by the OM barrier.
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Affiliation(s)
- William Ndugire
- Department of Chemistry, University of Massachusetts Lowell, One University Ave., Lowell, MA, 01854, USA
| | - N G Hasitha Raviranga
- Department of Chemistry, University of Massachusetts Lowell, One University Ave., Lowell, MA, 01854, USA
| | - Jingzhe Lao
- Department of Chemistry, University of Massachusetts Lowell, One University Ave., Lowell, MA, 01854, USA
| | - Olof Ramström
- Department of Chemistry, University of Massachusetts Lowell, One University Ave., Lowell, MA, 01854, USA
- Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar, SE-39182, Sweden
| | - Mingdi Yan
- Department of Chemistry, University of Massachusetts Lowell, One University Ave., Lowell, MA, 01854, USA
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Uddin F, Sohail M, Shaikh QH, Ahmed S, Khan S, Roulston K, McHugh TD. PCR and microarray analysis of AmpC and ESBLs producing Pseudomonas aeruginosa isolates from intensive care units. GENE REPORTS 2021. [DOI: 10.1016/j.genrep.2021.101178] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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Identification of the Pseudomonas aeruginosa O17 and O15 O-Specific Antigen Biosynthesis Loci Reveals an ABC Transporter-Dependent Synthesis Pathway and Mechanisms of Genetic Diversity. J Bacteriol 2020; 202:JB.00347-20. [PMID: 32690555 DOI: 10.1128/jb.00347-20] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/16/2020] [Accepted: 07/13/2020] [Indexed: 02/07/2023] Open
Abstract
Many bacterial cell surface glycans, such as the O antigen component of lipopolysaccharide (LPS), are produced via the so-called Wzx/Wzy- or ABC transporter-dependent pathways. O antigens are highly diverse polysaccharides that protect bacteria from their environment and engage in important host-pathogen interactions. The specific structure and composition of O antigens are the basis of classifying bacteria into O serotypes. In the opportunistic pathogen Pseudomonas aeruginosa, there are currently 20 known O-specific antigen (OSA) structures. The clusters of genes responsible for 18 of these O antigens have been identified, all of which follow the Wzx/Wzy-dependent pathway and are located at a common locus. In this study, we located the two unidentified O antigen biosynthesis clusters responsible for the synthesis of the O15 and the O17 OSA structures by analyzing published whole-genome sequence data. Intriguingly, these clusters were found outside the conserved OSA biosynthesis locus and were likely acquired through multiple horizontal gene transfer events. Based on data from knockout and overexpression studies, we determined that the synthesis of these O antigens follows an ABC transporter-dependent rather than a Wzx/Wzy-dependent pathway. In addition, we collected evidence to show that the O15 and O17 polysaccharide chain lengths are regulated by molecular rulers with distinct and variable domain architectures. The findings in this report are critical for a comprehensive understanding of O antigen biosynthesis in P. aeruginosa and provide a framework for future studies.IMPORTANCE P. aeruginosa is a problematic opportunistic pathogen that causes diseases in those with compromised host defenses, such as those suffering from cystic fibrosis. This bacterium produces a number of virulence factors, including a serotype-specific O antigen. Here, we identified and characterized the gene clusters that produce the O15 and O17 O antigens and show that they utilize a pathway for synthesis that is distinct from that of the 18 other known serotypes. We also provide evidence that these clusters have acquired mutations in specific biosynthesis genes and have undergone extensive horizontal gene transfer within the P. aeruginosa population. These findings expand on our understanding of O antigen biosynthesis in Gram-negative bacteria and the mechanisms that drive O antigen diversity.
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Kothari A, Kumar S, Omar BJ, Kiran K. Detection of extended-spectrum beta-lactamase (ESBL) production by disc diffusion method among Pseudomonas species from various clinical samples. J Family Med Prim Care 2020; 9:683-693. [PMID: 32318403 PMCID: PMC7114052 DOI: 10.4103/jfmpc.jfmpc_570_19] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2019] [Revised: 01/23/2020] [Accepted: 01/24/2020] [Indexed: 11/04/2022] Open
Abstract
Aim/Objectives This study was aimed to detect extended-spectrum beta-lactamase (ESBL) producing Pseudomonas species isolated from various clinical samples by phenotypic methods with their susceptibility testing. Materials and Methods Hundred Pseudomonas isolates were taken from various clinical samples of patients attending outpatient department (OPD) and inpatient department (IPD). Antimicrobial susceptibility test and ESBL detection were assessed using CLSI guidelines on Mueller Hinton agar. Results Out of 100 Pseudomonas isolates, 46 isolates were from female and 54 were from male patients. More cases of pseudomonal infection were in the age group between 46 and 60 years (34%), and 59% of Pseudomonas species were isolated from patients belongs to urban areas and the rest 41% were from rural. The isolates collected from OPD were 61% and rest 39% from IPD. Pseudomonas species showed maximum resistance to cephalosporin group of antibiotics and showed least resistance to imipenem, and showed 100% susceptibility to colistin. ESBL production was detected in 42% of total isolates. Conclusion The present study highlights that the Pseudomonas species remains an important cause of nosocomial infections. ESBL producing Pseudomonas species continue to be an important organism causing life-threatening infections. Multidrug resistance was seen in most of the strains. Resistance is developing even to combination of ceftazidime clavulanic acid. Resistance is developing to last resort of antibiotic, i.e. imipenem also. This gives the alarming signal for the future, making the therapeutic options more difficult. Strict infection control measures are to be taken to contain this so-called water and soil organisms as Pseudomonas.
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Affiliation(s)
- Ashish Kothari
- Department of Microbiology, All India Institute of Medical Science, Rishikesh, Uttarakhand, India
| | - Shailesh Kumar
- Department of Dentistry, All India Institute of Medical Science, Rishikesh, Uttarakhand, India
| | - Balram Ji Omar
- Department of Microbiology, All India Institute of Medical Science, Rishikesh, Uttarakhand, India
| | - Kamini Kiran
- Department of Pathology, All India Institute of Medical Science, Rishikesh, Uttarakhand, India
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Iovene MR, Pota V, Galdiero M, Corvino G, Lella FMD, Stelitano D, Passavanti MB, Pace MC, Alfieri A, Franco SD, Aurilio C, Sansone P, Niyas VKM, Fiore M. First Italian outbreak of VIM-producing Serratia marcescensin an adult polyvalent intensive care unit, August-October 2018: A case report and literature review. World J Clin Cases 2019. [DOI: 10.12998/wjcc.v7.i21.3518] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/04/2023] Open
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Iovene MR, Pota V, Galdiero M, Corvino G, Di Lella FM, Stelitano D, Passavanti MB, Pace MC, Alfieri A, Di Franco S, Aurilio C, Sansone P, Niyas VKM, Fiore M. First Italian outbreak of VIM-producing Serratia marcescens in an adult polyvalent intensive care unit, August-October 2018: A case report and literature review. World J Clin Cases 2019; 7:3535-3548. [PMID: 31750335 PMCID: PMC6854422 DOI: 10.12998/wjcc.v7.i21.3535] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/08/2019] [Revised: 05/15/2019] [Accepted: 07/27/2019] [Indexed: 02/05/2023] Open
Abstract
BACKGROUND Carbapenem-resistant Enterobacteriaceae has become a significant public health concern as hospital outbreaks are now being frequently reported and these organisms are becoming difficult to treat with the available antibiotics. CASE SUMMARY An outbreak of VIM-producing Serratia marcescens occurred over a period of 11 wk (August, 1 to October, 18) in patients admitted to the adult polyvalent intensive care unit of the University of Campania "Luigi Vanvitelli" located in Naples. Four episodes occurred in three patients (two patients infected, and one patient colonized). All the strains revealed the production of VIM. CONCLUSION After three decades of carbapenem antibiotics use, the emergence of carbapenem-resistance in Enterobacteriaceae has become a significant concern and a stricter control to preserve its clinical application is mandatory. This is, to our knowledge, the first outbreak of VIM-producing Serratia marcescens in Europe. Surveillance policies must be implemented to avoid future outbreaks.
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Affiliation(s)
- Maria Rosaria Iovene
- Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Vincenzo Pota
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Massimiliano Galdiero
- Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Giusy Corvino
- Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Federica Maria Di Lella
- Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Debora Stelitano
- Department of Experimental Medicine, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Maria Beatrice Passavanti
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Maria Caterina Pace
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Aniello Alfieri
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Sveva Di Franco
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Caterina Aurilio
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | - Pasquale Sansone
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
| | | | - Marco Fiore
- Department of Women, Child and General and Specialized Surgery, University of Campania "Luigi Vanvitelli", Naples 80138, Italy
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Wu B, Yang X, Yan M. Synthesis and Structure-Activity Relationship Study of Antimicrobial Auranofin against ESKAPE Pathogens. J Med Chem 2019; 62:7751-7768. [PMID: 31386365 PMCID: PMC7941214 DOI: 10.1021/acs.jmedchem.9b00550] [Citation(s) in RCA: 39] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023]
Abstract
Auranofin, an FDA-approved arthritis drug, has recently been repurposed as a potential antimicrobial agent; it performed well against many Gram-positive bacteria, including multidrug resistant strains. It is, however, inactive toward Gram-negative bacteria, for which we are in dire need of new therapies. In this work, 40 auranofin analogues were synthesized by varying the structures of the thiol and phosphine ligands, and their activities were tested against ESKAPE pathogens. The study identified compounds that exhibited bacterial inhibition (MIC) and killing (MBC) activities up to 65 folds higher than that of auranofin, making them effective against Gram-negative pathogens. Both thiol and the phosphine structures influence the activities of the analogues. The trimethylphosphine and triethylphosphine ligands gave the highest activities against Gram-negative and Gram-positive bacteria, respectively. Our SAR study revealed that the thiol ligand is also very important, the structure of which can modulate the activities of the AuI complexes for both Gram-negative and Gram-positive bacteria. Moreover, these analogues had mammalian cell toxicities either similar to or lower than that of auranofin.
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Affiliation(s)
- Bin Wu
- Department of Chemistry, The University of Massachusetts, Lowell, Massachusetts 01854, United States
| | - Xiaojian Yang
- Department of Chemistry, The University of Massachusetts, Lowell, Massachusetts 01854, United States
| | - Mingdi Yan
- Department of Chemistry, The University of Massachusetts, Lowell, Massachusetts 01854, United States
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13
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Dual β-lactam combination therapy for multi-drug resistant Pseudomonas aeruginosa infection: enhanced efficacy in vivo and comparison with monotherapies of penicillin-binding protein inhibition. Sci Rep 2019; 9:9098. [PMID: 31235728 PMCID: PMC6591303 DOI: 10.1038/s41598-019-45550-z] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/06/2019] [Accepted: 06/10/2019] [Indexed: 02/07/2023] Open
Abstract
The aim of the study was to determine the efficacy of dual β-lactam combination treatments derived from eight approved drugs against Galleria mellonella larvae infected with MDR strains of P. aeruginosa. Carbapenem-resistant P. aeruginosa NCTC 13437 and an unrelated clinical isolate were used to infect G. mellonella larvae and the efficacy of twenty-eight dual β-lactam combination therapies were compared to their constituent monotherapies. For the most potent combinations identified, penicillin-binding protein (PBP) inhibition profiles were measured and compared with each constituent antibiotic. Five of the dual β-lactam combinations resulted in greater than 70% survival of infected G. mellonella. Two combinations showed potent, enhanced efficacy versus both strains − ceftazidime + meropenem and aztreonam + meropenem. Comparison of PBP inhibition profiles revealed that the enhanced efficacy of these two dual β-lactam combinations could not be explained by more potent inhibition of PBPs or inhibition of a broader range of PBPs. A possible contribution to the enhanced efficacy of the combinations could be stimulation of innate immunity via increased haemocyte numbers compared to their constituent monotherapies. Combinations of β-lactam antibiotics show promise in overcoming MDR P. aeruginosa and are worthy of additional study and development.
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Prevalence of Extended Spectrum Betalactamase (ESBL) and Metallobetalactamase (MBL) Producing Pseudomonas aeruginosa and Acinetobacter baumannii Isolated from Various Clinical Samples. J Pathog 2018; 2018:6845985. [PMID: 30473888 PMCID: PMC6220378 DOI: 10.1155/2018/6845985] [Citation(s) in RCA: 30] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2018] [Accepted: 09/23/2018] [Indexed: 11/17/2022] Open
Abstract
This study was conducted with an objective to find the prevalence of extended spectrum betalactamase (ESBL) and metallobetalactamase (MBL) in P. aeruginosa and A. baumannii isolates obtained from various clinical samples. It was conducted in the Department of Microbiology, Adesh Institute of Medical Sciences and Research, Bathinda, over a period of two years from July 2014 to June 2016. Clinical specimens including urine, pus, blood, high vaginal swabs, respiratory samples, and various body fluids were processed and P. aeruginosa and A. baumannii isolates were identified by standard protocols. Antibiotic sensitivity testing for all isolates was done using Kirby-Bauer disc diffusion method. Disc potentiation test was performed to check ESBL and MBL production in these bacteria. Maximum ESBL positive isolates of P. aeruginosa were observed among pus samples and maximum MBL positive isolates were detected in tracheal aspirates. A. baumannii showed maximum positivity for ESBL and MBL production in endotracheal secretions. This study gives an alarming sign towards high prevalence of cephalosporin and carbapenem resistance due to production of extended spectrum betalactamases and metallobetalactamases, respectively. Early detection, stringent antibiotic policies, and compliance towards infection control practices are the best defenses against these organisms.
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Greenwood B, Meunier D, Hopkins KL, Pike R, Ivanov Z, Turton JF, Hill R, Woodford N, Livermore DM. Pseudomonas aeruginosa sequence type 357 with VEB extended-spectrum β-lactamases in the UK: relatedness and resistance. Int J Antimicrob Agents 2018; 52:301-302. [PMID: 29580931 DOI: 10.1016/j.ijantimicag.2018.03.013] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/21/2017] [Revised: 03/13/2018] [Accepted: 03/17/2018] [Indexed: 11/29/2022]
Affiliation(s)
- Beckie Greenwood
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Danièle Meunier
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK.
| | - Katie L Hopkins
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Rachel Pike
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Zdravko Ivanov
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Jane F Turton
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Robert Hill
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - Neil Woodford
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK
| | - David M Livermore
- Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit, National Infection Service, Public Health England, London, UK; Floor 2, Bob Champion Research & Educational Building, University of East Anglia, Norwich, Norfolk, UK
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Adjei CB, Govinden U, Moodley K, Essack SY. Molecular characterisation of multidrug-resistant Pseudomonas aeruginosa from a private hospital in Durban, South Africa. S Afr J Infect Dis 2017. [DOI: 10.1080/23120053.2017.1382090] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022] Open
Affiliation(s)
- Cosmos B. Adjei
- Antimicrobial Research Unit, University of KwaZulu-Natal, Westville, Durban, South Africa
| | - Usha Govinden
- Antimicrobial Research Unit, University of KwaZulu-Natal, Westville, Durban, South Africa
| | | | - Sabiha Y. Essack
- Antimicrobial Research Unit, University of KwaZulu-Natal, Westville, Durban, South Africa
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Laudy AE, Róg P, Smolińska-Król K, Ćmiel M, Słoczyńska A, Patzer J, Dzierżanowska D, Wolinowska R, Starościak B, Tyski S. Prevalence of ESBL-producing Pseudomonas aeruginosa isolates in Warsaw, Poland, detected by various phenotypic and genotypic methods. PLoS One 2017; 12:e0180121. [PMID: 28658322 PMCID: PMC5489192 DOI: 10.1371/journal.pone.0180121] [Citation(s) in RCA: 52] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2017] [Accepted: 06/11/2017] [Indexed: 01/17/2023] Open
Abstract
Knowledge of the prevalence of ESBL enzymes among P. aeruginosa strains compared to the Enterobacteraiceae family is limited. The phenotypic tests recommended by EUCAST for the detection of ESBL-producing Enterobacteriaceae are not always suited for P. aeruginosa strains. This is mainly due to the presence of other families of ESBLs in P. aeruginosa isolates more often than in Enterobacteriaceae, production of natural AmpC cephalosporinase and its overexpression, and co-production of metallo-β-lactamases. The aim of this study was to determine the occurrence of ESBLs in P. aeruginosa isolated from patients from hospitals in Warsaw, to evaluate the ESBL production of these isolates using currently available phenotypic tests, their modifications, multiplex PCR and molecular typing of ESBL-positive isolates by PFGE. Clinical isolates of P. aeruginosa were collected in 2000-2014 from four Warsaw hospitals. Based on the data obtained in this study, we suggest using three DDST methods with inhibitors, such as clavulanic acid, sulbactam and imipenem, to detect ESBL-producing P. aeruginosa strains. Depending on the appearance of the plates, we suggest a reduction in the distance between discs with antibiotics to 15 mm and the addition of boronic acid at 0.4 mg per disc. The analysed isolates carried genes encoding ESBL from the families VEB (69 isolates with VEB-9), GES (6 with GES-1, 1 GES-5, 5 GES-13 and 2 with GES-15), OXA-2 (12 with OXA-15, 1 OXA-141, 1 OXA-210, 1 OXA-543 and 1 with OXA-544) and OXA-10 (5 isolates with OXA-74 and one with OXA-142). The most important result of this study was the discovery of three new genes, blaGES-15, blaOXA-141 and blaOXA-142; their nucleotide sequences have been submitted to the NCBI GenBank. It is also very important to note that this is the first report on the epidemiological problem of VEB-9-producing bacterial strains, not only in Poland but also worldwide.
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Affiliation(s)
- Agnieszka E. Laudy
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
- * E-mail:
| | - Patrycja Róg
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
| | | | - Milena Ćmiel
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
| | - Alicja Słoczyńska
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
| | - Jan Patzer
- Department of Clinical Microbiology and Immunology, The Children’s Memorial Health Institute, Warsaw, Poland
| | - Danuta Dzierżanowska
- Department of Clinical Microbiology and Immunology, The Children’s Memorial Health Institute, Warsaw, Poland
| | - Renata Wolinowska
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
| | - Bohdan Starościak
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
| | - Stefan Tyski
- Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland
- Department of Antibiotics and Microbiology, National Medicines Institute, Warsaw, Poland
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Genetic Characterization of a blaVEB-2-Carrying Plasmid in Vibrio parahaemolyticus. Antimicrob Agents Chemother 2016; 60:6965-6968. [PMID: 27645248 DOI: 10.1128/aac.01749-16] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2016] [Accepted: 08/26/2016] [Indexed: 11/20/2022] Open
Abstract
This report describes the first detection of a blaVEB-2 gene in a Vibrio parahaemolyticus strain isolated from a shrimp sample. The blaVEB-2 gene was carried on a novel Inc-type plasmid that was likely to have originated from aquatic organisms, as indicated by a comparison with other known genetic elements in the GenBank database. However, the plasmid contains resistance elements usually harbored by members of the family Enterobacteriaceae, suggesting that gene transfer events occurred and contributed to the formation of this multidrug resistance-encoding plasmid.
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Jain S, Gaind R, Kothari C, Sehgal R, Shamweel A, Thukral SS, Chellani HK. VEB-1 extended-spectrum β-lactamase-producing multidrug-resistant Proteus mirabilis sepsis outbreak in a neonatal intensive care unit in India: clinical and diagnostic implications. JMM Case Rep 2016; 3:e005056. [PMID: 28348778 PMCID: PMC5330246 DOI: 10.1099/jmmcr.0.005056] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/03/2014] [Accepted: 06/21/2016] [Indexed: 12/03/2022] Open
Abstract
Introduction: Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae, multidrug-resistant (MDR) pathogens, are increasingly implicated in nosocomial outbreaksworldwide, particularly in neonatal intensive care units (NICUs). Proteus mirabilis is an uncommon nosocomial pathogen causing sepsis in neonates. Case Presentation: We report an outbreak of ESBL-positive MDR P. mirabilis sepsis involving five babies within 10 days in a NICU, which was promptly detected and managed. The aim of this study was to characterize the molecular mechanism of resistance to third-generation cephalosporins (3GCs) in the bacteria. Surveillance cultures were collected from health-care personnel (hand swabs, urine) and the surrounding patient-care environment. Ribotyping was performed to determine the clonality of the strain. Thirteen P. mirabilis were recovered from the blood cultures of the five babies and surveillance cultures. Twelve isolates were positive for the VEB-1 ESBL type, and were susceptible only to ciprofloxacin and carbapenems. There was an unusual phenotypic synergy observed between the 3GCs and imipenem/cefoxitin. The source of infection was traced to a contaminated multidose vial. The outbreak was associated with a high mortality (80 %). A change of empirical antibiotic policy to ciprofloxacin, with strict infection control measures, brought the outbreak to an abrupt end. Conclusion: This is believed to be the first report of a nosocomial outbreak of VEB-1 ESBL-producing P. mirabilis sepsis in neonates from India. The present report of infection due to VEB-1-producing P. mirabilis, an uncommon pathogen for an epidemic in a neonatal unit, highlights the growing significance of such Gram-negative bacteria as a cause of infections in newborns. Epidemic spread in a neonatal unit of an ESBL-producing Proteus species, which also had an intrinsically reduced susceptibility to imipenem, and resistance to colistin and tigecycline, can be a threatening situation and can result in high neonatal mortality unless recognized and controlled in a timely manner.
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Affiliation(s)
- Sarika Jain
- Department of Microbiology, VMMC and Safdarjang Hospital , New Delhi 110029 , India
| | - Rajni Gaind
- Department of Microbiology, VMMC and Safdarjang Hospital , New Delhi 110029 , India
| | - Charu Kothari
- Department of Microbiology, VMMC and Safdarjang Hospital , New Delhi 110029 , India
| | - Rachna Sehgal
- Department of Pediatrics, VMMC and Safdarjang Hospital , New Delhi 110029 , India
| | - A Shamweel
- Department of Microbiology, Vallabhbhai Patel Chest Institute , New Delhi , India
| | - S S Thukral
- Department of Microbiology, Vallabhbhai Patel Chest Institute , New Delhi , India
| | - Harish K Chellani
- Department of Pediatrics, VMMC and Safdarjang Hospital , New Delhi 110029 , India
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Diversity of Molecular Mechanisms Conferring Carbapenem Resistance to Pseudomonas aeruginosa Isolates from Saudi Arabia. CANADIAN JOURNAL OF INFECTIOUS DISEASES & MEDICAL MICROBIOLOGY 2016; 2016:4379686. [PMID: 27597874 PMCID: PMC4997076 DOI: 10.1155/2016/4379686] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Subscribe] [Scholar Register] [Received: 05/17/2016] [Accepted: 07/19/2016] [Indexed: 01/07/2023]
Abstract
Background. This study described various molecular and epidemiological characters determining antibiotic resistance patterns in Pseudomonas aeruginosa isolates. Methods. A total of 34 carbapenem-resistant P. aeruginosa clinical isolates were isolated from samples collected at a tertiary hospital in Riyadh, Saudi Arabia, from January to December 2011. Susceptibility testing, serotyping, molecular characterization of carbapenem resistance, and pulsed-field gel electrophoresis (PFGE) were performed. Results. All isolates were resistant to ceftazidime, and more than half were highly resistant (minimum inhibitory concentration (MIC) > 256 mg/L). Fifteen isolates had MIC values ≥64 mg/L for any of the carbapenems examined. Vietnamese extended-spectrum β-lactamase (VEB-1) (n = 16/34) and oxacillinase (OXA-10) (n = 14/34) were the most prevalent extended-spectrum β-lactamase and penicillinase, respectively. Verona imipenemase (VIM-1, VIM-2, VIM-4, VIM-11, and VIM-28) and imipenemase (IMP-7) variants were found in metallo-β-lactamase producers. A decrease in outer membrane porin gene (oprD) expression was seen in nine isolates, and an increase in efflux pump gene (MexAB) expression was detected in five isolates. Six serotypes (O:1, O:4, O:7, O:10, O:11, and O:15) were found among the 34 isolates. The predominant serotype was O:11 (16 isolates), followed by O:15 (nine isolates). PFGE analysis of the 34 carbapenem-resistant P. aeruginosa isolates revealed 14 different pulsotypes. Conclusions. These results revealed diverse mechanisms conferring carbapenem resistance to P. aeruginosa isolates from Saudi Arabia.
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El Zowalaty ME, Gyetvai B. Effectiveness of Antipseudomonal Antibiotics and Mechanisms of Multidrug Resistance in Pseudomonas aeruginosa. Pol J Microbiol 2016. [DOI: 10.5604/17331331.1197272] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
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Multidrug resistant, blaVEB positive Pseudomonas aeruginosa causing high mortality among haematology patients. Pathology 2015; 46:650-2. [PMID: 25393259 DOI: 10.1097/pat.0000000000000177] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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Wright LL, Turton JF, Hopkins KL, Livermore DM, Woodford N. Genetic environment of metallo-β-lactamase genes in Pseudomonas aeruginosa isolates from the UK. J Antimicrob Chemother 2015; 70:3250-8. [PMID: 26318194 DOI: 10.1093/jac/dkv263] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/01/2015] [Accepted: 07/29/2015] [Indexed: 01/01/2023] Open
Abstract
OBJECTIVES We sought to characterize the genetic environment of blaVIM and blaIMP genes in Pseudomonas aeruginosa isolates from the UK; these included members of six previously described prevalent complexes, A-F, which correspond to international 'high-risk clones', along with diverse strains. METHODS Metallo-β-lactamase (MBL)-encoding class 1 integrons were amplified by PCR from 218 P. aeruginosa isolates producing VIM-type (n = 196) or IMP-type (n = 22) enzymes, referred from UK hospital laboratories between 2003 and 2012. The variable regions of selected integrons were sequenced using a primer walking method. RESULTS One-hundred-and-nineteen isolates had an MBL-encoding integron with the 3' conserved sequence (3'CS), 65 had Tn5090-like 3' regions and 17 had the sul1 gene, but lacked the qacEΔ1 gene; the 3' region could not be amplified using any primer combinations for the remaining 17 isolates. Six integron profiles were each seen in more than five isolates. Predominant integron types were seen amongst isolates belonging to STs 111, 233, 654/964 and 773 (complexes A, C, D and F, respectively), whereas diverse integron profiles were seen in isolates belonging to ST235 (complex B) and ST357 (complex E). CONCLUSIONS In UK P. aeruginosa isolates, MBL genes occur in diverse class 1 integron structures, though commonly with 3' regions containing the classical 3'CS or Tn5090-like regions. Four of the six main clonal complexes, referred from multiple laboratories, carried a predominant integron type, whereas the remaining two had more diverse types.
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Affiliation(s)
- Laura L Wright
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, 61 Colindale Avenue, London NW9 5EQ, UK Norwich Medical School, University of East Anglia, Norwich, Norfolk NR4 7TJ, UK
| | - Jane F Turton
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, 61 Colindale Avenue, London NW9 5EQ, UK
| | - Katie L Hopkins
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, 61 Colindale Avenue, London NW9 5EQ, UK
| | - David M Livermore
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, 61 Colindale Avenue, London NW9 5EQ, UK Norwich Medical School, University of East Anglia, Norwich, Norfolk NR4 7TJ, UK
| | - Neil Woodford
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, 61 Colindale Avenue, London NW9 5EQ, UK
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Hong DJ, Bae IK, Jang IH, Jeong SH, Kang HK, Lee K. Epidemiology and Characteristics of Metallo-β-Lactamase-Producing Pseudomonas aeruginosa. Infect Chemother 2015; 47:81-97. [PMID: 26157586 PMCID: PMC4495280 DOI: 10.3947/ic.2015.47.2.81] [Citation(s) in RCA: 165] [Impact Index Per Article: 16.5] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2015] [Indexed: 12/18/2022] Open
Abstract
Metallo-β-lactamase-producing Pseudomonas aeruginosa (MPPA) is an important nosocomial pathogen that shows resistance to all β-lactam antibiotics except monobactams. There are various types of metallo-β-lactamases (MBLs) in carbapenem-resistant P. aeruginosa including Imipenemase (IMP), Verona integron-encoded metallo-β-lactamase (VIM), Sao Paulo metallo-β-lactamase (SPM), Germany imipenemase (GIM), New Delhi metallo-β-lactamase (NDM), Florence imipenemase (FIM). Each MBL gene is located on specific genetic elements including integrons, transposons, plasmids, or on the chromosome, in which they carry genes encoding determinants of resistance to carbapenems and other antibiotics, conferring multidrug resistance to P. aeruginosa. In addition, these genetic elements are transferable to other Gram-negative species, increasing the antimicrobial resistance rate and complicating the treatment of infected patients. Therefore, it is essential to understand the epidemiology, resistance mechanism, and molecular characteristics of MPPA for infection control and prevention of a possible global health crisis. Here, we highlight the characteristics of MPPA.
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Affiliation(s)
- Duck Jin Hong
- Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea
| | - Il Kwon Bae
- Department of Dental Hygiene, Silla University, Busan, Korea
| | - In-Ho Jang
- Department of Biomedical Laboratory Science, College of Health Sciences, Sangji University, Wonju, Korea
| | - Seok Hoon Jeong
- Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea
| | - Hyun-Kyung Kang
- Department of Dental Hygiene, Silla University, Busan, Korea
| | - Kyungwon Lee
- Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea
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Adamson DH, Krikstopaityte V, Coote PJ. Enhanced efficacy of putative efflux pump inhibitor/antibiotic combination treatments versus MDR strains ofPseudomonas aeruginosain aGalleria mellonella in vivoinfection model. J Antimicrob Chemother 2015; 70:2271-8. [DOI: 10.1093/jac/dkv111] [Citation(s) in RCA: 34] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2015] [Accepted: 04/01/2015] [Indexed: 11/14/2022] Open
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Potron A, Poirel L, Nordmann P. Emerging broad-spectrum resistance in Pseudomonas aeruginosa and Acinetobacter baumannii: Mechanisms and epidemiology. Int J Antimicrob Agents 2015; 45:568-85. [PMID: 25857949 DOI: 10.1016/j.ijantimicag.2015.03.001] [Citation(s) in RCA: 484] [Impact Index Per Article: 48.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2015] [Accepted: 03/05/2015] [Indexed: 02/07/2023]
Abstract
Multidrug resistance is quite common among non-fermenting Gram-negative rods, in particular among clinically relevant species including Pseudomonas aeruginosa and Acinetobacter baumannii. These bacterial species, which are mainly nosocomial pathogens, possess a diversity of resistance mechanisms that may lead to multidrug or even pandrug resistance. Extended-spectrum β-lactamases (ESBLs) conferring resistance to broad-spectrum cephalosporins, carbapenemases conferring resistance to carbapenems, and 16S rRNA methylases conferring resistance to all clinically relevant aminoglycosides are the most important causes of concern. Concomitant resistance to fluoroquinolones, polymyxins (colistin) and tigecycline may lead to pandrug resistance. The most important mechanisms of resistance in P. aeruginosa and A. baumannii and their most recent dissemination worldwide are detailed here.
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Affiliation(s)
- Anaïs Potron
- Laboratoire de Bactériologie, Faculté de Médecine-Pharmacie, Centre Hospitalier Régional Universitaire, Université de Franche-Comté, Besançon, France
| | - Laurent Poirel
- Emerging Antibiotic Resistance Medical and Molecular Microbiology Unit, Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, Switzerland.
| | - Patrice Nordmann
- Emerging Antibiotic Resistance Medical and Molecular Microbiology Unit, Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, Switzerland; HFR - Hôpital Cantonal de Fribourg, Fribourg, Switzerland
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Bokaeian M, Shahraki Zahedani S, Soltanian Bajgiran M, Ansari Moghaddam A. Frequency of PER, VEB, SHV, TEM and CTX-M Genes in Resistant Strains of Pseudomonas aeruginosa Producing Extended Spectrum β-Lactamases. Jundishapur J Microbiol 2014; 8:e13783. [PMID: 25789123 PMCID: PMC4350043 DOI: 10.5812/jjm.13783] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/27/2013] [Revised: 01/18/2014] [Accepted: 01/21/2014] [Indexed: 11/16/2022] Open
Abstract
Background: Pseudomonas aeruginosa is the most common pathogen causing nosocomial infections. Resistance of P. aeruginosa strains to broad-spectrum cephalosporins may be mediated by extended-spectrum β-lactamases (ESBLs). Objectives: We intended to investigate the prevalence of ESBLs and antimicrobial susceptibilities of P. aeruginosa isolated from patients in Zahedan, Iran. Materials and Methods: In this cross-sectional study, during 2012–2013, 116 P. aeruginosa isolates were collected from a teaching hospital in Zahedan, Iran. Susceptibility to eight antimicrobial agents was carried out by disk diffusion method. The ESBL producing strains were detected by combination disk test (CDT). ESBL positive isolates as well as other isolates showing minimum inhibitory concentrations (MICs) ≥ 4 μg/mL for ceftazidime, cefotaxime, ceftriaxone and aztreonam, were screened for the presence of the genes encoding blaTEM, blaSHV, blaPER-1 and blaVEB-1, by polymerase chain reaction (PCR). Results: Ciprofloxacin and piperacillin were the most efficient antipseudomonal agents. The results disclosed that 19 (16.37%) of the isolates were multidrug resistant and 8 (6.89%) were ESBL-positive. Of the 116 isolates, 30 (25.86%) were resistant to at least one of the antibiotics ceftazidime, ceftriaxone, cefotaxime or aztreonam and among these 30 (100%), 4 (13.3%), 2 (6.6%) and 2 (6.6%), amplified blaTEM, blaVEB-1, blaPER-1 and blaSHV, respectively. From the 30 TEM-positive isolates, 22 were ESBL-negative. Sequencing of the ESBL genes verified the accuracy of the PCR products. Conclusions: According to our results, blaTEM-116 was the most frequent isolated ESBL gene among the P. aeruginosa strains isolated from patients.
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Affiliation(s)
- Mohmmad Bokaeian
- Department of Microbiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, IR Iran
- Research Center for Infectious Diseases and Tropical Medicine, Zahedan University of Medical Sciences, Zahedan, IR Iran
| | - Shahram Shahraki Zahedani
- Department of Microbiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, IR Iran
| | - Morteza Soltanian Bajgiran
- Department of Microbiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, IR Iran
- Corresponding author: Morteza Soltanian Bajgiran, , Department of Microbiology, Zahedan University of Medical Sciences, Zahedan, IR Iran. Tel: +98-5413414558, Fax: +98-5413425723, E-mail:
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Yezli S, Shibl AM, Memish ZA. The molecular basis of β-lactamase production in Gram-negative bacteria from Saudi Arabia. J Med Microbiol 2014; 64:127-136. [PMID: 25418734 DOI: 10.1099/jmm.0.077834-0] [Citation(s) in RCA: 30] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
Resistance to β-lactams among Gram-negative bacteria is a worldwide issue. Increased prevalence of extended-spectrum β-lactamase (ESBL)-producers and the dissemination of carbapenem-resistance genes are particularly concerning. ESBL-producing strains are common in the Kingdom of Saudi Arabia, particularly among the Enterobacteriaceae, and carbapenem resistance is on the increase, especially among the non-fermenters. β-lactamase production is a major mechanism of resistance to these agents and although β-lactamase-producing strains have been documented in the Kingdom, relatively few reports characterized the molecular basis of this production. Nevertheless, available data suggest that CTX-M (CTX-M-15 in particular) is the predominant ESBL in the Enterobacteriaceae, with SHV also being prevalent in Klebsiella pneumoniae. Carbapenem resistance in the latter is mainly due to OXA-48 and NDM-1. In Pseudomonas aeruginosa, VEB-like enzymes are the most common ESBLs, and VIM is the prevalent metallo-β-lactamase. OXA-10 extended-spectrum enzymes are also frequent. PER and GES ESBLs have been reported in Acinetobacter baumannii, and oxacillinases (OXA-23 in particular) are the dominant carbapanamases in this species.
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Affiliation(s)
- Saber Yezli
- Saudi Ministry of Health, Riyadh, Saudi Arabia
| | | | - Ziad A Memish
- College of Medicine, Alfaisal University, Riyadh, Saudi Arabia.,Saudi Ministry of Health, Riyadh, Saudi Arabia
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Wright LL, Turton JF, Livermore DM, Hopkins KL, Woodford N. Dominance of international 'high-risk clones' among metallo-β-lactamase-producing Pseudomonas aeruginosa in the UK. J Antimicrob Chemother 2014; 70:103-10. [PMID: 25182064 DOI: 10.1093/jac/dku339] [Citation(s) in RCA: 66] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023] Open
Abstract
OBJECTIVES Carbapenem-resistant isolates of Pseudomonas aeruginosa producing metallo-β-lactamases (MBLs) are increasingly reported worldwide and often belong to particular 'high-risk clones'. This study aimed to characterize a comprehensive collection of MBL-producing P. aeruginosa isolates referred to the UK national reference laboratory from multiple UK laboratories over a 10 year period. METHODS Isolates were referred to the UK national reference laboratory between 2003 and 2012 for investigation of resistance mechanisms and/or outbreaks. MBL genes were detected by PCR. Typing was carried out by nine-locus variable-number tandem repeat (VNTR) analysis and MLST. RESULTS MBL-producing P. aeruginosa isolates were referred from 267 source patients and 89 UK laboratories. The most common isolation sites were urine (24%), respiratory (18%), wounds (17%) and blood (13%). VIM-type MBLs predominated (91% of all MBLs found), but a few IMP- and NDM-type enzymes were also identified. Diverse VNTR types were seen, but 86% of isolates belonged to six major complexes. MLST of representative isolates from each complex showed that they corresponded to STs 111, 233, 235, 357, 654 and 773, respectively. Isolates belonging to these complexes were received from between 9 and 25 UK referring laboratories each. CONCLUSIONS The incidence of MBL-producing P. aeruginosa is increasing in the UK. The majority of these isolates belong to several 'high-risk clones', which have been previously reported internationally as host clones of MBLs.
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Affiliation(s)
- Laura L Wright
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, Colindale, London NW9 5EQ, UK Norwich Medical School, University of East Anglia, Norwich, Norfolk NR4 7TJ, UK
| | - Jane F Turton
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, Colindale, London NW9 5EQ, UK
| | - David M Livermore
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, Colindale, London NW9 5EQ, UK Norwich Medical School, University of East Anglia, Norwich, Norfolk NR4 7TJ, UK
| | - Katie L Hopkins
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, Colindale, London NW9 5EQ, UK
| | - Neil Woodford
- Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, Colindale, London NW9 5EQ, UK
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Krezdorn J, Adams S, Coote PJ. A Galleria mellonella infection model reveals double and triple antibiotic combination therapies with enhanced efficacy versus a multidrug-resistant strain of Pseudomonas aeruginosa. J Med Microbiol 2014; 63:945-955. [PMID: 24928215 DOI: 10.1099/jmm.0.074245-0] [Citation(s) in RCA: 47] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022] Open
Abstract
The aim of this study was to compare the inhibitory effect of antibiotic combinations in vitro with efficacy in Galleria mellonella larvae in vivo to identify efficacious combinations that target Pseudomonas aeruginosa. P. aeruginosa NCTC 13437, a multidrug-resistant strain resistant to β-lactams and aminoglycosides, was used. Susceptibility to cefotaxime, piperacillin, meropenem, amikacin, levofloxacin and colistin alone, or in dual or triple combinations, was measured in vitro via a 24 h time-kill assay. In vitro results were then compared with the efficacy of the same dual or triple antibiotic combinations versus G. mellonella larvae infected with P. aeruginosa. G. mellonella haemolymph burden of P. aeruginosa was determined over 96 h post-infection and treatment with the most potent combination therapies. Many dual and triple combinations of antibiotics displayed synergistic inhibition of multidrug-resistant P. aeruginosa in vitro. There was little correlation between combinations that were synergistic in vitro and those that showed enhanced efficacy in vivo versus infected G. mellonella larvae. The most potent dual and triple combinations in vivo were cefotaxime plus piperacillin, and meropenem plus piperacillin and amikacin, respectively. Fewer combinations were found to offer enhanced therapeutic benefit in vivo compared with in vitro. The therapeutic benefit arising from treatment with antibiotic combinations in vivo correlated with reduced larval burden of P. aeruginosa. This study has identified antibiotic combinations that merit further investigation for their clinical potential and has demonstrated the utility of using G. mellonella to screen for novel antibiotic treatments that demonstrate efficacy in vivo.
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Affiliation(s)
- Jessica Krezdorn
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, UK
| | - Sophie Adams
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, UK
| | - Peter J Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews KY16 9ST, UK
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Bahmani N, Ramazanzadeh R. Detection of SHV type Extended-Spectrum B-lactamase and Risk Factors in Pseudomonas aeruginosa Clinical Isolates. Pak J Med Sci 2013; 29:788-92. [PMID: 24353629 PMCID: PMC3809287 DOI: 10.12669/pjms.293.3263] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2012] [Revised: 04/22/2013] [Accepted: 04/23/2013] [Indexed: 11/15/2022] Open
Abstract
OBJECTIVE Pseudomonas aeruginosa is one of the most important causes of nosocomial infections and can acquire resistant to many antimicrobials, including β-lactams. The aim of this study was to detect the prevalence of SHV type extended-spectrum beta-lactamase (ESBL), antimicrobial resistance patterns of the P. aeroginusa and risk factors in hospitalized patients in two teaching hospitals in Sanandaj, Iran. METHODOLOGY 123 P. aeruginosa were isolated from various clinical specimens. All samples were prepared for double-disk synergy test on the isolates for detection of ESBL. SHV was confirmed by PCR method. Risk factors were evaluated for infection due to P. aeruginosa. RESULTS The incidence of multiple drug resistance (MDR) in P. aeroginusa isolates was 3.85%. The prevalence of ESBL-SHV gene was 10.57%. Days of hospitalization (OR=14.34 CI95% 2.87-25.8), ICU hospitalization (OR=3.4 CI95% 1.24- 9.29), presence of catheter (OR=3.63 CI 95% 1.34-9.84), use of antimicrobials within previous two weeks (OR=5.51 CI95% 1.85-16.43) and use of ventilator (OR=3.7557 CI95%1.29-9) were risk factors for Pseudomonas nosocomial infection SHV positive ESBL. CONCLUSION In this study Prevalence of ESBL, SHV gene and MDR in P. aeroginosa infection was lower than the prevalence reported from other studies in Iran and this indicated appropriate antimicrobial managements strategies and infection control. In addition, our research data indicate that risk factors such as use of ventilator, use of antimicrobials and ICU hospitalization can be effective in managing Pseudomonas infection.
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Affiliation(s)
- Nasrin Bahmani
- Nasrin Bahmani, MS, Microbiology Department, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj-Iran
| | - Rashid Ramazanzadeh
- Rashid Ramazanzadeh, Associated Professor, Cellular and Molecular Research Center, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj-Iran
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Hill L, Veli N, Coote PJ. Evaluation of Galleria mellonella larvae for measuring the efficacy and pharmacokinetics of antibiotic therapies against Pseudomonas aeruginosa infection. Int J Antimicrob Agents 2013; 43:254-61. [PMID: 24361354 DOI: 10.1016/j.ijantimicag.2013.11.001] [Citation(s) in RCA: 90] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/17/2013] [Revised: 11/06/2013] [Accepted: 11/07/2013] [Indexed: 11/18/2022]
Abstract
The aim of this study was to determine whether Galleria mellonella larvae can be used (i) as an in vivo infection model for Pseudomonas aeruginosa and (ii) for evaluating the pharmacokinetics and efficacy of antipseudomonal antibiotics. Two strains of P. aeruginosa were employed, NCTC 10662 (antibiotic-susceptible) and NCTC 13437 (multidrug-resistant). Larvae were infected with increasing doses of either P. aeruginosa strain to investigate the effect of inoculum size on survival. Subsequently, infected larvae were treated with a range of antibiotics to examine whether these agents were effective against P. aeruginosa infection in vivo and whether the efficacy of these drugs matched the known susceptibilities of each bacterial strain. Larval burden of P. aeruginosa was also determined after infection and treatment with cefotaxime. Pharmacokinetic properties of the antibiotics tested were measured using a well diffusion assay to determine the concentration of antibiotics in larval haemolymph over time. Galleria mellonella larvae were sensitive to P. aeruginosa infection, and increasing inoculum doses of live cells resulted in greater larval mortality. Heat-killed bacteria had no detrimental effect on survival. Antibiotic efficacy against P. aeruginosa-infected G. mellonella correlated with the measured in vitro sensitivities of the two strains tested. The therapeutic benefit arising from administration of cefotaxime correlated with a reduced burden of bacteria present in the haemolymph. There was a clear correlation between measured antibiotic pharmacokinetics and the therapeutic effect. This study strongly supports future application of the G. mellonella infection model to initial studies of novel antipseudomonal treatments.
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Affiliation(s)
- Lucy Hill
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Neyme Veli
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK
| | - Peter J Coote
- Biomedical Sciences Research Complex, School of Biology, University of St Andrews, The North Haugh, St Andrews, Fife KY16 9ST, UK.
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Al-Agamy MH, Shibl AM, Tawfik AF, Elkhizzi NA, Livermore DM. Extended-spectrum and metallo-beta-lactamases among ceftazidime-resistantPseudomonas aeruginosain Riyadh, Saudi Arabia. J Chemother 2013; 24:97-100. [DOI: 10.1179/1120009x12z.00000000015] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/31/2022]
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Begum S, Salam MA, Alam KF, Begum N, Hassan P, Haq JA. Detection of extended spectrum β-lactamase in Pseudomonas spp. isolated from two tertiary care hospitals in Bangladesh. BMC Res Notes 2013; 6:7. [PMID: 23289861 PMCID: PMC3543721 DOI: 10.1186/1756-0500-6-7] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2012] [Accepted: 01/04/2013] [Indexed: 05/27/2023] Open
Abstract
Background Extended spectrum ß-lactamases (ESBLs) represent a major group of lactamases responsible for resistance, mostly produced by gram-negative bacteria, to newer generations of ß-lactam drugs currently being identified in large numbers worldwide. The present study was undertaken to see the frequency of ESBL producing Pseudomonas spp. isolated from six hundred clinical specimens (wound, pus, aural, urine, sputum, throat and other swabs) collected over a period of three years from two tertiary care hospitals in Bangladesh. Findings Aerobic bacterial culture was performed on aseptically collected swabs and only growth of Pseudomonas was considered for further species identification and ESBL production along with serotyping of Pseudomonas aeruginosa. Antimicrobial susceptibility testing was carried out using the Kirby-Bauer agar diffusion method and ESBL production was detected on Mueller Hinton agar by double-disk synergy technique using Amoxicillin-Clavulanic acid with Ceftazidime, Cefotaxime, Ceftriaxone and Aztreonam. Culture yielded 120 Pseudomonas spp. and 82 of them were biochemically characterized for species. Pseudomonas aeruginosa was found to be the predominant (90.2%) species. Of 82 isolates tested for ESBL, 31 (37.8%) were ESBL positive with 29 (93.5%) as Pseudomonas aeruginosa, the remaining 2 (6.5%) were Stenotrophomonas maltophilia and Ralstonia pickettii. Antibiogram revealed Imipenem as the most effective drug (93.3%) among all antimicrobials used against Pseudomonas spp. followed by Aminoglycosides (63.7%). Conclusion ESBL producing Pseudomonas spp. was found to be a frequent isolate from two tertiary care hospitals in Bangladesh, showing limited susceptibility to antimicrobials and decreased susceptibility to Imipenem in particular, which is a matter of great concern.
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Affiliation(s)
- Shahanara Begum
- Department of Microbiology, Green Life Medical College, Dhaka, Bangladesh
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Tawfik AF, Shibl AM, Aljohi MA, Altammami MA, Al-Agamy MH. Distribution of Ambler class A, B and D β-lactamases among Pseudomonas aeruginosa isolates. Burns 2012; 38:855-60. [DOI: 10.1016/j.burns.2012.01.005] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2011] [Revised: 11/13/2011] [Accepted: 01/02/2012] [Indexed: 11/16/2022]
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Polotto M, Casella T, de Lucca Oliveira MG, Rúbio FG, Nogueira ML, de Almeida MT, Nogueira MC. Detection of P. aeruginosa harboring bla CTX-M-2, bla GES-1 and bla GES-5, bla IMP-1 and bla SPM-1 causing infections in Brazilian tertiary-care hospital. BMC Infect Dis 2012; 12:176. [PMID: 22863113 PMCID: PMC3512492 DOI: 10.1186/1471-2334-12-176] [Citation(s) in RCA: 60] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/13/2012] [Accepted: 07/24/2012] [Indexed: 12/27/2022] Open
Abstract
Background Nosocomial infections caused by Pseudomonas aeruginosa presenting resistance to beta-lactam drugs are one of the most challenging targets for antimicrobial therapy, leading to substantial increase in mortality rates in hospitals worldwide. In this context, P. aeruginosa harboring acquired mechanisms of resistance, such as production of metallo-beta-lactamase (MBLs) and extended-spectrum beta-lactamases (ESBLs) have the highest clinical impact. Hence, this study was designed to investigate the presence of genes codifying for MBLs and ESBLs among carbapenem resistant P. aeruginosa isolated in a Brazilian 720-bed teaching tertiary care hospital. Methods Fifty-six carbapenem-resistant P. aeruginosa strains were evaluated for the presence of MBL and ESBL genes. Strains presenting MBL and/or ESBL genes were submitted to pulsed-field gel electrophoresis for genetic similarity evaluation. Results Despite the carbapenem resistance, genes for MBLs (blaSPM-1 or blaIMP-1) were detected in only 26.7% of isolates. Genes encoding ESBLs were detected in 23.2% of isolates. The blaCTX-M-2 was the most prevalent ESBL gene (19.6%), followed by blaGES-1 and blaGES-5 detected in one isolate each. In all isolates presenting MBL phenotype by double-disc synergy test (DDST), the blaSPM-1 or blaIMP-1 genes were detected. In addition, blaIMP-1 was also detected in three isolates which did not display any MBL phenotype. These isolates also presented the blaCTX-M-2 gene. The co-existence of blaCTX-M-2 with blaIMP-1 is presently reported for the first time, as like as co-existence of blaGES-1 with blaIMP-1. Conclusions In this study MBLs production was not the major mechanism of resistance to carbapenems, suggesting the occurrence of multidrug efflux pumps, reduction in porin channels and production of other beta-lactamases. The detection of blaCTX-M-2,blaGES-1 and blaGES-5 reflects the recent emergence of ESBLs among antimicrobial resistant P. aeruginosa and the extraordinary ability presented by this pathogen to acquire multiple resistance mechanisms. These findings raise the concern about the future of antimicrobial therapy and the capability of clinical laboratories to detect resistant strains, since simultaneous production of MBLs and ESBLs is known to promote further complexity in phenotypic detection. Occurrence of intra-hospital clonal dissemination enhances the necessity of better observance of infection control practices.
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Affiliation(s)
- Milena Polotto
- Laboratório de Microbiologia, Departamento de Doenças Dermatológicas, Infecciosas e Parasitárias, São José do Rio Preto, SP, Brazil
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Jabalameli F, Mirsalehian A, Sotoudeh N, Jabalameli L, Aligholi M, Khoramian B, Taherikalani M, Emaneini M. Multiple-locus variable number of tandem repeats (VNTR) fingerprinting (MLVF) and antibacterial resistance profiles of extended spectrum beta lactamase (ESBL) producing Pseudomonas aeruginosa among burnt patients in Tehran. Burns 2011; 37:1202-7. [DOI: 10.1016/j.burns.2011.05.012] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2010] [Revised: 04/30/2011] [Accepted: 05/25/2011] [Indexed: 11/27/2022]
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Abstract
Pseudomonas aeruginosa is intrinsically resistant to a variety of antimicrobials and can develop resistance during anti-pseudomonal chemotherapy both of which compromise treatment of infections caused by this organism. Resistance to multiple classes of antimicrobials (multidrug resistance) in particular is increasingly common in P. aeruginosa, with a number of reports of pan-resistant isolates treatable with a single agent, colistin. Acquired resistance in this organism is multifactorial and attributable to chromosomal mutations and the acquisition of resistance genes via horizontal gene transfer. Mutational changes impacting resistance include upregulation of multidrug efflux systems to promote antimicrobial expulsion, derepression of ampC, AmpC alterations that expand the enzyme's substrate specificity (i.e., extended-spectrum AmpC), alterations to outer membrane permeability to limit antimicrobial entry and alterations to antimicrobial targets. Acquired mechanisms contributing to resistance in P. aeruginosa include β-lactamases, notably the extended-spectrum β-lactamases and the carbapenemases that hydrolyze most β-lactams, aminoglycoside-modifying enzymes, and 16S rRNA methylases that provide high-level pan-aminoglycoside resistance. The organism's propensity to grow in vivo as antimicrobial-tolerant biofilms and the occurrence of hypermutator strains that yield antimicrobial resistant mutants at higher frequency also compromise anti-pseudomonal chemotherapy. With limited therapeutic options and increasing resistance will the untreatable P. aeruginosa infection soon be upon us?
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Affiliation(s)
- Keith Poole
- Department of Microbiology and Immunology, Queen's University Kingston, ON, Canada
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40
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Cornaglia G, Giamarellou H, Rossolini GM. Metallo-β-lactamases: a last frontier for β-lactams? THE LANCET. INFECTIOUS DISEASES 2011; 11:381-93. [PMID: 21530894 DOI: 10.1016/s1473-3099(11)70056-1] [Citation(s) in RCA: 496] [Impact Index Per Article: 35.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
Metallo-β-lactamases are resistance determinants of increasing clinical relevance in Gram-negative bacteria. Because of their broad range, potent carbapenemase activity and resistance to inhibitors, these enzymes can confer resistance to almost all β-lactams. Since the 1990s, several metallo-β-lactamases encoded by mobile DNA have emerged in important Gram-negative pathogens (ie, in Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii). Some of these enzymes (eg, VIM-1 and NDM-1) have been involved in the recent crisis resulting from the international dissemination of carbapenem-resistant Klebsiella pneumoniae and other enterobacteria. Although substantial knowledge about the molecular biology and genetics of metallo-β-lactamases is available, epidemiological data are inconsistent and clinical experience is still lacking; therefore, several unsolved or debatable issues remain about the management of infections caused by producers of metallo-β-lactamase. The spread of metallo-β-lactamases presents a major challenge both for treatment of individual patients and for policies of infection control, exposing the substantial unpreparedness of public health structures in facing up to this emergency.
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Affiliation(s)
- Giuseppe Cornaglia
- Department of Pathology and Diagnostics, University of Verona, Verona, Italy.
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41
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Poole K. Pseudomonas aeruginosa: resistance to the max. Front Microbiol 2011; 2:65. [PMID: 21747788 PMCID: PMC3128976 DOI: 10.3389/fmicb.2011.00065] [Citation(s) in RCA: 580] [Impact Index Per Article: 41.4] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/28/2011] [Accepted: 03/24/2011] [Indexed: 01/04/2023] Open
Abstract
Pseudomonas aeruginosa is intrinsically resistant to a variety of antimicrobials and can develop resistance during anti-pseudomonal chemotherapy both of which compromise treatment of infections caused by this organism. Resistance to multiple classes of antimicrobials (multidrug resistance) in particular is increasingly common in P. aeruginosa, with a number of reports of pan-resistant isolates treatable with a single agent, colistin. Acquired resistance in this organism is multifactorial and attributable to chromosomal mutations and the acquisition of resistance genes via horizontal gene transfer. Mutational changes impacting resistance include upregulation of multidrug efflux systems to promote antimicrobial expulsion, derepression of ampC, AmpC alterations that expand the enzyme's substrate specificity (i.e., extended-spectrum AmpC), alterations to outer membrane permeability to limit antimicrobial entry and alterations to antimicrobial targets. Acquired mechanisms contributing to resistance in P. aeruginosa include β-lactamases, notably the extended-spectrum β-lactamases and the carbapenemases that hydrolyze most β-lactams, aminoglycoside-modifying enzymes, and 16S rRNA methylases that provide high-level pan-aminoglycoside resistance. The organism's propensity to grow in vivo as antimicrobial-tolerant biofilms and the occurrence of hypermutator strains that yield antimicrobial resistant mutants at higher frequency also compromise anti-pseudomonal chemotherapy. With limited therapeutic options and increasing resistance will the untreatable P. aeruginosa infection soon be upon us?
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Affiliation(s)
- Keith Poole
- Department of Microbiology and Immunology, Queen's University Kingston, ON, Canada
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42
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Zhao WH, Hu ZQ. β-Lactamases identified in clinical isolates ofPseudomonas aeruginosa. Crit Rev Microbiol 2010; 36:245-58. [DOI: 10.3109/1040841x.2010.481763] [Citation(s) in RCA: 72] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
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Glupczynski Y, Bogaerts P, Deplano A, Berhin C, Huang TD, Van Eldere J, Rodriguez-Villalobos H. Detection and characterization of class A extended-spectrum- -lactamase-producing Pseudomonas aeruginosa isolates in Belgian hospitals. J Antimicrob Chemother 2010; 65:866-71. [DOI: 10.1093/jac/dkq048] [Citation(s) in RCA: 59] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
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Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms. Clin Microbiol Rev 2010; 22:582-610. [PMID: 19822890 DOI: 10.1128/cmr.00040-09] [Citation(s) in RCA: 1218] [Impact Index Per Article: 81.2] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Treatment of infectious diseases becomes more challenging with each passing year. This is especially true for infections caused by the opportunistic pathogen Pseudomonas aeruginosa, with its ability to rapidly develop resistance to multiple classes of antibiotics. Although the import of resistance mechanisms on mobile genetic elements is always a concern, the most difficult challenge we face with P. aeruginosa is its ability to rapidly develop resistance during the course of treating an infection. The chromosomally encoded AmpC cephalosporinase, the outer membrane porin OprD, and the multidrug efflux pumps are particularly relevant to this therapeutic challenge. The discussion presented in this review highlights the clinical significance of these chromosomally encoded resistance mechanisms, as well as the complex mechanisms/pathways by which P. aeruginosa regulates their expression. Although a great deal of knowledge has been gained toward understanding the regulation of AmpC, OprD, and efflux pumps in P. aeruginosa, it is clear that we have much to learn about how this resourceful pathogen coregulates different resistance mechanisms to overcome the antibacterial challenges it faces.
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Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms. Clin Microbiol Rev 2009. [PMID: 19822890 DOI: 10.1128/cmr.00040-09.] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Treatment of infectious diseases becomes more challenging with each passing year. This is especially true for infections caused by the opportunistic pathogen Pseudomonas aeruginosa, with its ability to rapidly develop resistance to multiple classes of antibiotics. Although the import of resistance mechanisms on mobile genetic elements is always a concern, the most difficult challenge we face with P. aeruginosa is its ability to rapidly develop resistance during the course of treating an infection. The chromosomally encoded AmpC cephalosporinase, the outer membrane porin OprD, and the multidrug efflux pumps are particularly relevant to this therapeutic challenge. The discussion presented in this review highlights the clinical significance of these chromosomally encoded resistance mechanisms, as well as the complex mechanisms/pathways by which P. aeruginosa regulates their expression. Although a great deal of knowledge has been gained toward understanding the regulation of AmpC, OprD, and efflux pumps in P. aeruginosa, it is clear that we have much to learn about how this resourceful pathogen coregulates different resistance mechanisms to overcome the antibacterial challenges it faces.
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A blaVEB-1 variant, blaVEB-6, associated with repeated elements in a complex genetic structure. Antimicrob Agents Chemother 2009; 53:1693-7. [PMID: 19139283 DOI: 10.1128/aac.01313-08] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
bla(VEB-6) was found on the Proteus mirabilis chromosome in a context similar to those of bla(VEB-1a) and bla(VEB-1b), in a truncated gene cassette flanked by 135-bp elements and duplications of the 3'-conserved segment of class 1 integrons. A linked aacA4-aadB-dfrA1-orfC cassette array includes components of Tn1331, illustrating the complex mosaicism of multiresistance regions.
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