Anti-Mutant Efficacy of Combination Therapy with Doripenem and Levofloxacin: In Vitro Model Studies with Pseudomonas Aeruginosa

2021 ◽  
Vol 66 (3-4) ◽  
pp. 12-17
Author(s):  
M. V. Golikova ◽  
E. N. Strukova ◽  
K. N. Alieva ◽  
A. V. Filimonova ◽  
Yu. A. Portnoy ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Combination Therapy ◽  
Bacterial Resistance ◽  
Combined Therapy ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Model Studies ◽  
Development Of Resistance ◽  
Mutant Prevention Concentration

Relevance. The tendency to a decrease in sensitivity of bacterial agents to old antibiotics, as well as the slowdown in creation of new medications, dictate the need to develop effective approaches to combat bacterial resistance.Aim. Evaluation of the applicability of a pharmacokinetically-based approach to predicting anti-mutant effectiveness of combined therapy with doripenem and levofloxacin against gram-negative bacteria Pseudomonas aeruginosa.Material and methods. A collection strain of Pseudomonas aeruginosa was used in the study. The values of MPC (mutant prevention concentration) of the combination of doripenem and levofloxacin were evaluated at a ratio of their concentrations equal to therapeutic ratios of the area under the pharmacokinetic curve in the in vitro dynamic model. 5-day treatments with clinical doses of doripenem and levofloxacin individually and in combination were simulated. Bacteria-containing medium was sampled during the experiments and plated on agar media containing 2MIC of each antibiotic.Results. The MPCs of doripenem and levofloxacin decreased 4 times when used in combination compared to MPC values when used separately. P.aeruginosa population was enriched with resistant mutants during monotherapy with each medication; the number of the bacteria did not decrease or even increased by the end of observation period. The use of doripenem/levofloxacin combination completely prevented development of resistance to both drugs in P.aeruginosa. The observed anti-mutant effect of antibiotic combination was consistent with higher (compared to monotherapy) values of the time during which the concentration of the antibiotic exceeded MPC (T>MPC).Conclusion. The anti-mutant effectiveness of combined therapy with doripenem and levofloxacin increased with the decrease in the values of MPC of antibiotics when used simultaneously, which consequently led to the increase in the values of T>MPC. Obtained results confirm the applicability of a pharmacokinetically-based approach to the estimation of MPC of combined antibiotics for predicting anti-mutant effectiveness of combination therapy in the treatment of infections caused by gram-negative bacteria.

scholarly journals Development of Resistance in Wild-Type and Hypermutable Pseudomonas aeruginosa Strains Exposed to Clinical Pharmacokinetic Profiles of Meropenem and Ceftazidime Simulated In Vitro

2007 ◽  
Vol 51 (10) ◽  
pp. 3642-3649 ◽  
Author(s):  
Beate Henrichfreise ◽  
Irith Wiegand ◽  
Ingeborg Luhmer-Becker ◽  
Bernd Wiedemann
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Parent Strain ◽  
Resistance Mechanisms ◽  
In Vitro Models ◽  
Wild Type ◽  
Resistance Development ◽  
Development Of Resistance ◽  
Pharmacokinetic Profiles ◽  
Mutant Prevention Concentration

ABSTRACT In this study we investigated the interplay of antibiotic pharmacokinetic profiles and the development of mutation-mediated resistance in wild-type and hypermutable Pseudomonas aeruginosa strains. We used in vitro models simulating profiles of the commonly used therapeutic drugs meropenem and ceftazidime, two agents with high levels of antipseudomonal activity said to have different potentials for stimulating resistance development. During ceftazidime treatment of the wild-type strain (PAO1), fully resistant mutants overproducing AmpC were selected rapidly and they completely replaced wild-type cells in the population. During treatment with meropenem, mutants of PAO1 were not selected as rapidly and showed only intermediate resistance due to the loss of OprD. These mutants also replaced the parent strain in the population. During the treatment of the mutator P. aeruginosa strain with meropenem, the slowly selected mutants did not accumulate several resistance mechanisms but only lost OprD and did not completely replace the parent strain in the population. Our results indicate that the commonly used dosing regimens for meropenem and ceftazidime cannot avoid the selection of mutants of wild-type and hypermutable P. aeruginosa strains. For the treatment outcome, including the prevention of resistance development, it would be beneficial for the antibiotic concentration to remain above the mutant prevention concentration for a longer period of time than it does in present regimens.

Antibacterial Activity and Chemical Composition of the Essential Oil of Grammosciadium platycarpum Boiss. from Iran

2005 ◽  
Vol 60 (1-2) ◽  
pp. 30-34 ◽  
Author(s):  
Ali Sonboli ◽  
Fereshteh Eftekhar ◽  
Morteza Yousefzadi ◽  
Mohammad Reza Kanani
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibacterial Activity ◽  
Chemical Composition ◽  
Essential Oil ◽  
Essential Oils ◽  
Antibacterial Activities ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Two Samples

The chemical composition of the essential oils obtained from two samples (GP1 and GP2) of Grammosciadium platycarpum Boiss. was analyzed by GC and GC-MS. The analysis of the oils resulted in the identification of twenty-two constituents. Linalool (79.0% - GP1, 81.8% - GP2) and limonene (10.0%, 5.8%) were found to be the major components, respectively. The in vitro antibacterial activities of these oils and their main compounds against seven Gram-positive and Gram-negative bacteria were investigated. The results exhibited that the total oils and their major components possess strong to moderate activities against all the tested bacteria except for Pseudomonas aeruginosa.

scholarly journals Exposure to the Gram-Negative Bacteria Pseudomonas aeruginosa Influences the Lung Dendritic Cell Population Signature by Interfering With CD103 Expression

2021 ◽  
Vol 11 ◽  
Author(s):  
Julyanne Brassard ◽  
Joanny Roy ◽  
Anne-Marie Lemay ◽  
Marie-Josée Beaulieu ◽  
Emilie Bernatchez ◽  
...  
Keyword(s):  
Immune Response ◽  
Pseudomonas Aeruginosa ◽  
Intracellular Localization ◽  
Receptor Internalization ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Gm Csf ◽  
Dendritic Cell Population ◽  
The Impact

Lung dendritic cells (DCs) are divided into two major populations, which include CD103+XCR1+ cDC1s and CD11b+Sirpα+ cDC2s. The maintenance of their relative proportions is dynamic and lung inflammation, such as caused by exposure to lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, can have a significant impact on the local cDC signature. Alterations in the lung cDC signature could modify the capacity of the immune system to respond to various pathogens. We consequently aimed to assess the impact of the Gram-negative bacteria Pseudomonas aeruginosa on lung cDC1 and cDC2 populations, and to identify the mechanisms leading to alterations in cDC populations. We observed that exposure to P. aeruginosa decreased the proportions of CD103+XCR1+ cDC1s, while increasing that of CD11b+ DCs. We identified two potential mechanisms involved in this modulation of lung cDC populations. First, we observed an increase in bone marrow pre-DC IRF4 expression suggesting a higher propensity of pre-DCs to differentiate towards the cDC2 lineage. This observation was combined with a reduced capacity of lung XCR1+ DC1s to express CD103. In vitro, we demonstrated that GM-CSF-induced CD103 expression on cDCs depends on GM-CSF receptor internalization and RUNX1 activity. Furthermore, we observed that cDCs stimulation with LPS or P. aeruginosa reduced the proportions of intracellular GM-CSF receptor and decreased RUNX1 mRNA expression. Altogether, these results suggest that alterations in GM-CSF receptor intracellular localization and RUNX1 signaling could be involved in the reduced CD103 expression on cDC1 in response to P. aeruginosa. To verify whether the capacity of cDCs to express CD103 following P. aeruginosa exposure impacts the immune response, WT and Cd103-/- mice were exposed to P. aeruginosa. Lack of CD103 expression led to an increase in the number of neutrophils in the airways, suggesting that lack of CD103 expression on cDC1s could favor the innate immune response to this bacterium.

scholarly journals In vitro activity of the siderophore cephalosporin, cefiderocol, against molecularly characterized, carbapenem-non-susceptible Gram-negative bacteria from Europe

2020 ◽  
Vol 2 (3) ◽  
Author(s):  
Christopher Longshaw ◽  
Davide Manissero ◽  
Masakatsu Tsuji ◽  
Roger Echols ◽  
Yoshinori Yamano
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antimicrobial Agents ◽  
Similar Proportion ◽  
Gram Negative Bacteria ◽  
In Vitro Activity ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
Gene Profiles ◽  
Carbapenemase Gene

Abstract Objectives Many carbapenem-resistant (CR) Gram-negative (GN) pathogens exhibit MDR, meaning few therapeutic options are available for CR-GN infections. Cefiderocol, a siderophore cephalosporin, has demonstrated in vitro efficacy against CR-GN bacteria. In the SIDERO-CR-2014–2016 surveillance study, European clinical isolates comprising carbapenem-non-susceptible (CarbNS) Enterobacterales and MDR non-fermenters were tested against cefiderocol and comparators. Methods Cefiderocol MICs were determined using iron-depleted CAMHB, and comparators using CAMHB, per recommended CLSI methodology. Carbapenemase gene profiles were determined using PCR. Results Isolates (N = 870) from 23 European countries comprised CarbNS Enterobacterales (n = 457), MDR Pseudomonas aeruginosa (n = 177) and MDR Acinetobacter baumannii (n = 236). The most common carbapenemases were KPC (52%), OXA-48-like (19%), VIM (14%) and NDM (8%) in Enterobacterales, VIM (41%) in P. aeruginosa and OXA-23-like (57%) and OXA-24/40-like (37%) in A. baumannii. Most carbapenemase-producing isolates (65%) co-carried ESBLs. Approximately half of P. aeruginosa isolates were negative for carbapenemases, compared with 10% of Enterobacterales and 3% of A. baumannii. A similar proportion of Enterobacterales were susceptible to cefiderocol (81.6%; 79.0% of VIM producers; 51.4% of NDM producers; based on EUCAST breakpoint values) compared with comparator antimicrobial agents, including colistin (76.4%; 93.5% of VIM producers; 78.4% of NDM producers) and ceftazidime/avibactam (76.6%; 1.6% of VIM producers; 2.7% of NDM producers). Of P. aeruginosa isolates, 98.3% were susceptible to cefiderocol (100% of VIM producers), similar to colistin (100%). Against A. baumannii, 94.9% had cefiderocol MIC ≤2 mg/L and 93.6% of isolates were susceptible to colistin. Conclusions Cefiderocol demonstrated potent activity against CarbNS and MDR GN bacteria, including non-fermenters and a wide variety of MBL- and serine-β-lactamase-producing strains.

scholarly journals Multiple Mechanisms of the Synthesized Antimicrobial Peptide TS against Gram-Negative Bacteria for High Efficacy Antibacterial Action In Vivo

Molecules ◽  
2020 ◽  
Vol 26 (1) ◽  
pp. 60
Author(s):  
Rui Zhang ◽  
Xiaobo Fan ◽  
Xinglu Jiang ◽  
Mingyuan Zou ◽  
Han Xiao ◽  
...  
Keyword(s):  
Antimicrobial Peptide ◽  
Bacterial Infections ◽  
Bacterial Resistance ◽  
Divalent Cations ◽  
Resistant Bacteria ◽  
Antibacterial Drug ◽  
Gram Negative Bacteria ◽  
Gram Negative

The emergence of drug-resistant bacteria emphasizes the urgent need for novel antibiotics. The antimicrobial peptide TS shows extensive antibacterial activity in vitro and in vivo, especially in gram-negative bacteria; however, its antibacterial mechanism is unclear. Here, we find that TS without hemolytic activity disrupts the integrity of the outer bacterial cell membrane by displacing divalent cations and competitively binding lipopolysaccharides. In addition, the antimicrobial peptide TS can inhibit and kill E. coli by disintegrating the bacteria from within by interacting with bacterial DNA. Thus, antimicrobial peptide TS’s multiple antibacterial mechanisms may not easily induce bacterial resistance, suggesting use as an antibacterial drug to be for combating bacterial infections in the future.

scholarly journals In VitroActivity of Ceftazidime-Avibactam against 338 Molecularly Characterized Gentamicin-Nonsusceptible Gram-Negative Clinical Isolates Obtained from Patients in Canadian Hospitals

2015 ◽  
Vol 59 (6) ◽  
pp. 3623-3626 ◽  
Author(s):  
Andrew J. Denisuik ◽  
James A. Karlowsky ◽  
Tyler Denisuik ◽  
Wright W. Nichols ◽  
Thomas A. Keating ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
16S Rrna ◽  
Klebsiella Pneumoniae ◽  
Gram Negative Bacteria ◽  
Aminoglycoside Resistance ◽  
Single Strain ◽  
Gram Negative ◽  
Content Type ◽  
16S Rrna Methylase

ABSTRACTThe mechanism of aminoglycoside resistance among 338 gentamicin-nonsusceptible Gram-negative bacteria (207Enterobacteriaceaeand 131Pseudomonas aeruginosa) was assessed, and thein vitroactivity of ceftazidime-avibactam against these isolates was determined. Aminoglycoside-modifying enzymes were detected in 91.8% ofEnterobacteriaceaeand 13.7% ofP. aeruginosaisolates. A single strain ofKlebsiella pneumoniaeharbored a 16S rRNA methylase (ArmA). The ceftazidime-avibactam MIC90values were 0.5 μg/ml (MIC, ≤8 μg/ml for 100% of isolates) and 16 μg/ml (MIC, ≤8 μg/ml for 87.8% of isolates) against gentamicin-nonsusceptibleEnterobacteriaceaeandP. aeruginosaisolates, respectively.

scholarly journals Outer Membrane Targeting of Pseudomonas aeruginosa Proteins Shows Variable Dependence on the Components of Bam and Lol Machineries

mBio ◽  
2011 ◽  
Vol 2 (6) ◽  
Author(s):  
Hanh H. Hoang ◽  
Nicholas N. Nickerson ◽  
Vincent T. Lee ◽  
Anastasia Kazimirova ◽  
Mohamed Chami ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Outer Membrane ◽  
Protein Translocation ◽  
Lipid Vesicles ◽  
Structural Features ◽  
Gram Negative Bacteria ◽  
Variable Effect ◽  
Gram Negative ◽  
Content Type

ABSTRACT In Gram-negative bacteria, the Lol and Bam machineries direct the targeting of lipidated and nonlipidated proteins, respectively, to the outer membrane (OM). Using Pseudomonas aeruginosa strains with depleted levels of specific Bam and Lol proteins, we demonstrated a variable dependence of different OM proteins on these targeting pathways. Reduction in the level of BamA significantly affected the ability of the β-barrel membrane protein OprF to localize to the OM, while the targeting of three secretins that are functionally related OM proteins was less affected (PilQ and PscC) or not at all affected (XcpQ). Depletion of LolB affected all lipoproteins examined and had a variable effect on the nonlipidated proteins. While the levels of OprF, PilQ, and PscC were significantly reduced by LolB depletion, XcpQ was unaffected and was correctly localized to the OM. These results suggest that certain β-barrel proteins such as OprF primarily utilize the complete Bam machinery. The Lol machinery participates in the OM targeting of secretins to variable degrees, likely through its involvement in the assembly of lipidated Bam components. XcpQ, but not PilQ or PscC, was shown to assemble spontaneously into liposomes as multimers. This work raises the possibility that there is a gradient of utilization of Bam and Lol insertion and targeting machineries. Structural features of individual proteins, including their β-barrel content, may determine the propensity of these proteins for folding (or misfolding) during periplasmic transit and OM insertion, thereby influencing the extent of utilization of the Bam targeting machinery, respectively. IMPORTANCE Targeting of lipidated and nonlipidated proteins to the outer membrane (OM) compartment in Gram-negative bacteria involves the transfer across the periplasm utilizing the Lol and Bam machineries, respectively. We show that depletion of Bam and Lol components in Pseudomonas aeruginosa does not lead to a general OM protein translocation defect, but the severity (and therefore, Lol and Bam dependence), varies with individual proteins. XcpQ, the secretin component of the type II secretion apparatus, is translocated into the OM without the assistance of Bam or Lol machineries. The hypothesis that XcpQ, after secretion across the cytoplasmic membrane, does not utilize the OM targeting machineries was supported by demonstrating that in vitro-synthesized XcpQ (but not the other P. aeruginosa secretins) can spontaneously incorporate into lipid vesicles. Therefore, the requirement for ancillary factors appears to be, in certain instances, dictated by the intrinsic properties of individual OM proteins, conceivably reflecting their propensities to misfold during periplasmic transit.

scholarly journals Evaluation of the Revogene Carba C Assay for Detection and Differentiation of Carbapenemase-Producing Gram-Negative Bacteria

2020 ◽  
Vol 58 (4) ◽  
Author(s):  
Delphine Girlich ◽  
Marine Laguide ◽  
Laurent Dortet ◽  
Thierry Naas
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Nucleic Acid ◽  
Sensitivity And Specificity ◽  
Multidrug Resistant ◽  
Gram Negative Bacteria ◽  
Microbiology Laboratory ◽  
Bacterial Host ◽  
Gram Negative ◽  
Content Type

ABSTRACT The Revogene Carba C assay (formerly GenePOC Carba assay) is a multiplex nucleic acid-based in vitro diagnostic test intended for the detection of carbapenemase-producing Enterobacterales (CPE) from cultured colonies. This assay was evaluated directly on colonies of 118 well-characterized Enterobacterales with reduced susceptibility to carbapenems and on 49 multidrug-resistant (MDR) Pseudomonas aeruginosa and 40 MDR Acinetobacter baumannii isolates. The Revogene Carba C assay’s performance was high, as it was able to detect the five major carbapenemases (NDM, VIM, IMP, KPC, and OXA-48). In Enterobacterales, sensitivity and specificity were 100%. When extrapolating the results to the French CPE epidemiology between 2012 and 2018, this assay would have detected 99.28% of the 9,624 CPE isolates sent to the French NRC, missing 69 CPE isolates (2 GES-5, 10 OXA-23, 2 TMB-1, 1 SME-4, 53 IMI, and 1 FRI). The overall sensitivity and specificity for CP P. aeruginosa were 93.7 and 100%, respectively, as two rare IMP variants (IMP-31 and -46) were not detected. Extrapolating these results to the French epidemiology of CP P. aeruginosa in 2017, 93.3% would have been identified, missing only 1 DIM and 10 GES variants. The Revogene Carba C assay accurately identified the targeted carbapenemase genes in A. baumannii, but when extrapolating these results to the French CP A. baumannii epidemiology of 2017, only 12.50% of them could be detected, as OXA-23 is the most prevalent carbapenemase in CP A. baumannii. The Revogene Carba C assay showed excellent sensitivity and specificity for the five most common carbapenemases regardless of the bacterial host. It is well adapted to the CPE and CP P. aeruginosa epidemiology of many countries worldwide, which makes it suitable for use in the routine microbiology laboratory, with a time to result of ca. 85 min for eight isolates simultaneously.

scholarly journals The Analysis Of Antibiotic Consumption And Bacterial Resistance As An Indicator Of Their Proper Use At The Urology Department In The Health Centre “Studenica” Kraljevo

2015 ◽  
Vol 16 (2) ◽  
pp. 135-141
Author(s):  
Andriana Bukonjić ◽  
Srđan Stefanović
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Bacterial Resistance ◽  
Health Centre ◽  
Antibiotic Consumption ◽  
Resistance Rate ◽  
Gram Negative Bacteria ◽  
Financial Assets ◽  
Bacterial Strains ◽  
Gram Negative

Abstract The objective of the study was to analyze antibiotic consumption and determine bacterial resistance rates as an indicator of the rational utilization of this drug group at the urology department in the Health Centre “Studenica” Kraljevo. Over a two-year period, the average antibiotic consumption was 104.55 DDD/100BD. Of the total financial assets used for medical treatment, the antibiotic group JO1 had a share of 49.52% in 2011 and 47.53% in 2012. Antibacterial drugs from a group of β-lactamic antibiotics were consumed most commonly, at 54.02% (2011) and 43.44% (2012). First-generation cephalosporins, quinolones and aminoglycosides were the most frequently used drug groups, while cephalexin was the antibiotic with the highest individual consumption. E. coli was the most frequently isolated bacterium in 2011, while in 2012, Klebsiella pneumoniae was the most frequently isolated bacterium. The total bacterial resistance both in 2011 and 2012 was above 50%. Gram-negative bacteria showed a higher resistance rate (2011, 59.3%; 2012, 58.9%) than Gram-positive bacteria (2011, 44.4%; 2012, 40.6%). Klebsiella pneumoniae was the bacterium with the highest resistance (75.3%) in 2011, while in 2012, there was a resistance increase in Pseudomonas aeruginosa (71.4%), especially to carbapenems. A correlation was determined between the consumption of individual antibiotics and bacterial strain resistance in 2011 (r=0.433, p=0.044) and in 2012 (r=0.478, p=0.024). The high resistance rate shown in the bacterial strains, which was correlated with antibiotic consumption, as well as the great financial assets used for this group of drugs suggest the necessity for the rationalization of their utilization. Empirical therapy with Gram-negative bacteria should be based on carbapenems utilization, except with Pseudomonas aeruginosa, where piperacillin/tazobactam should be used.

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