Concentrations Of Tigecycline Research Articles

CusS-CusR Two-Component System Mediates Tigecycline Resistance in Carbapenem-Resistant Klebsiella pneumoniae.

BackgroundThe increase in carbapenem-resistant Klebsiella pneumoniae (CRKP), especially the emergence of tigecycline-resistant K. pneumoniae (KP), is a serious public health concern. However, the underlying mechanism of tigecycline resistance is unclear. In this study, we evaluated the role of the CusS-CusR two-component system (TCS), which is associated with copper/silver resistance, in tigecycline resistance in CRKP.MethodsFollowing the in vitro evolution of tigecycline-resistant KP, the minimum inhibitory concentrations of tigecycline were determined using the micro-broth dilution method. RNA sequencing and data analysis were performed to identify differentially expressed genes. Quantitative PCR (qPCR) was performed to verify the genes of interest. Genes associated with tigecycline resistance, such as ramR, tex (T), and tet (A), were detected by PCR, and then mutants were confirmed by sequencing. Additionally, the efflux pump-associated genes soxS, oqxA, oqxB, acrE, and acrF were also analyzed by qPCR. CusR was deleted and complemented by the suicide vector pKO3-Km plasmid and pGEM-T-easy plasmid, respectively.ResultsNine strains of KP were evaluated in our study. Strains A2 and A3 were evolved from A1, B2, and B3 were evolved from B1, and C2 and C3 were evolved from C1. The tigecycline minimum inhibitory concentration for A1, B1, and C1 was 0.5 μg/mL; that for A2, B2, and C3 was 16.0 μg/mL; and that for A3, B3, and C3 was 32.0 μg/mL. RNA-sequencing and qPCR confirmed that the differentially expressed genes cusE, cusS, cusR, cusC, cusF, cusB, and cusA showed higher expression in C2 and C3 than in C1. Genes related to the efflux pump AcrAB-TolC showed higher expression in B2 and B3 than in B1. No mutants of ramR, tex (T), or tet (A) were detected. SoxS, oqxA, oqxB, acrE, and acrF did not show increased expression in any group. After deletion and complementation of cusR among C3, the MIC of tigecycline decreased to 4 μg/mL, and then recovered to 32 μg/mL. The expression of cusFBCA, correspondingly decreased and increased significantly.ConclusionIn addition to its primary function in resistance to copper/silver, the CusS-CusR two-component system is associated with CRKP resistance to tigecycline.

Synthesis, characterization, and evaluation of antimicrobial activity of novel Chitosan/Tigecycline composite

With the increase of antibiotic resistance currently available at an alarming rate, new improved films of Chitosan (CS) have been achieved with different levels of Tigecycline (TIGE) using traditional casting technology. The structural, optical, and morphological characterizations of the prepared (CS/TIGE) composites were investigated using XRD, FT-IR, UV–visible, and FE-SEM. Also, the antibacterial characterization was investigated. XRD pattern shows a semi-crystalline peak at 2θ = 21° begins to appear at Cs/TIGE2 concentration and its intensity increase with increasing TIGE concentration which indicate the interaction between TIGE and Cs. FT-IR reveals conservation of the distinctive bands with change in both intensity and position that attributed to increase the ratios of TIGE in Chitosan. UV–vis spectra confirmed the interaction of the CS and the drug TIGE. FE-SEM photos confirm the increasing of TIGE particles on the surface of CS/TIGE films by increasing the ratios of TIGE. The antimicrobial effect of Chitosan and five different ratios of Chitosan with TIGE have been identified against four different bacterial species, two gram-negative (Escherichia coli, Bacillus subtilis) and two gram-positive bacteria (Staphylococcus aureus, Pseudomonas aeruginosa). The obtained results indicated that the composite Cs/TIGE can be used in many antimicrobial applications because of its greatly antimicrobial activity.

Five-year Pan-European, longitudinal surveillance of Clostridium difficile ribotype prevalence and antimicrobial resistance: the extended ClosER study

Clostridium difficile infection (CDI) has been primarily treated with metronidazole or vancomycin. High recurrence rates, the emergence of epidemic PCR ribotypes (RTs) and the introduction of fidaxomicin in Europe in 2011 necessitate surveillance of antimicrobial resistance and CDI epidemiology. The ClosER study monitored antimicrobial susceptibility and geographical distribution of C. difficile RTs pre- and post-fidaxomicin introduction. From 2011 to 2016, 28 European countries submitted isolates or faecal samples for determination of PCR ribotype, toxin status and minimal inhibitory concentrations (MICs) of metronidazole, vancomycin, rifampicin, fidaxomicin, moxifloxacin, clindamycin, imipenem, chloramphenicol and tigecycline. RT diversity scores for each country were calculated and mean MIC results used to generate cumulative resistant scores (CRSs) for each isolate and country. From 40 sites, 3499 isolates were analysed, of which 95% (3338/3499) were toxin positive. The most common of the 264 RTs isolated was RT027 (mean prevalence 11.4%); however, RT prevalence varied greatly between countries and between years. The fidaxomicin geometric mean MIC for years 1–5 was 0.04 mg/L; only one fidaxomicin-resistant isolate (RT344) was submitted (MIC ≥ 4 mg/L). Metronidazole and vancomycin geometric mean MICs were 0.46 mg/L and 0.70 mg/L, respectively. Of prevalent RTs, RT027, RT017 and RT012 demonstrated resistance or reduced susceptibility to multiple antimicrobials. RT diversity was inversely correlated with mean CRS for individual countries (Pearson coefficient r = − 0.57). Overall, C. difficile RT prevalence remained stable in 2011–2016. Fidaxomicin susceptibility, including in RT027, was maintained post-introduction. Reduced ribotype diversity in individual countries was associated with increased antimicrobial resistance.

Mechanisms and phenotypic consequences of acquisition of tigecycline resistance by Stenotrophomonas maltophilia

To elucidate the potential mutation-driven mechanisms involved in the acquisition of tigecycline resistance by the opportunistic pathogen Stenotrophomonas maltophilia. The mutational trajectories and their effects on bacterial fitness, as well as cross-resistance and/or collateral susceptibility to other antibiotics, were also addressed. S. maltophilia populations were submitted to experimental evolution in the presence of increasing concentrations of tigecycline for 30 days. The genetic mechanisms involved in the acquisition of tigecycline resistance were determined by WGS. Resistance was evaluated by performing MIC assays. Fitness of the evolved populations and individual clones was assessed by measurement of the maximum growth rates. All the tigecycline-evolved populations attained high-level resistance to tigecycline following different mutational trajectories, yet with some common elements. Among the mechanisms involved in low susceptibility to tigecycline, mutations in the SmeDEF efflux pump negative regulator smeT, changes in proteins involved in the biogenesis of the ribosome and modifications in the LPS biosynthesis pathway seem to play a major role. Besides tigecycline resistance, the evolved populations presented cross-resistance to other antibiotics, such as aztreonam and quinolones, and they were hypersusceptible to fosfomycin, suggesting a possible combination treatment. Further, we found that the selected resistance mechanisms impose a relevant fitness cost when bacteria grow in the absence of antibiotic. Mutational resistance to tigecycline was easily selected during exposure to this antibiotic. However, the fitness cost may compromise the maintenance of S. maltophilia tigecycline-resistant populations in the absence of antibiotic.

Developmental dynamics of antibiotic resistome in aerobic biofilm microbiota treating wastewater under stepwise increasing tigecycline concentrations

This study aimed to investigate the impact of tigecycline, the third generation tetracycline, on the antibiotic resistance development in environmental microbiota. Two biological contact oxidation reactors containing aerobic biofilm microbiota were constructed, one of which was constantly fed with synthetic wastewater spiked with increasing concentrations of tigecycline (0 to 25 mg/L) under a hydrolytic retention time of 24 h. Over a period of 636 days, chemical oxygen demand removal over 90% and complete nitrification were achieved for both the control and tigecycline-exposed reactors, and effluent tigecycline concentrations in the tigecycline-exposed system were always <0.051 mg/L. Significant increases (p < 0.01) in resistome abundance and resistant bacteria ratio were detected at a tigecycline dose of 10 and 25 mg/L, respectively, revealed by metagenomic sequencing and culture-based method. The increase of resistome in the tigecycline system was mainly attributed to the enrichment of tetX, one cooperative tetracycline degrading gene. Partial canonical correspondence analysis showed that the change of resistome was mainly driven by bacterial community shift (vertical pathway). Network and genome binning analyses further suggested that the proliferation of Flavobacterium harboring tetX contributed to a relatively low community-wide resistance development in the aerobic biofilm microbiota under tigecycline selection by reducing the antibiotic concentration. This work provides scientific bases for the management and evaluation of the resistance risk induced by this novel antibiotic.

Evaluation of Plazomicin, Tigecycline, and Meropenem Pharmacodynamic Exposure against Carbapenem-Resistant Enterobacteriaceae in Patients with Bloodstream Infection or Hospital-Acquired/Ventilator-Associated Pneumonia from the CARE Study (ACHN-490-007).

IntroductionCARE was a Phase 3, randomized study evaluating the efficacy and safety of plazomicin-based combination therapy compared with colistin-based combination therapy for the treatment of patients with bloodstream infections or hospital-acquired/ventilator-associated pneumonia due to carbapenem-resistant Enterobacteriaceae (CRE). Adjunctive therapies included either tigecycline or meropenem. We sought to understand the contribution of tigecycline and meropenem to plazomicin-treated-patient outcomes by determining their observed pharmacodynamic exposures against baseline pathogens.MethodsBlood samples collected for plazomicin therapeutic monitoring were assayed for tigecycline and meropenem concentrations. Population pharmacokinetic models were constructed for each antibiotic. Using the individual Bayesian posterior or a covariate-based model, concentration time profiles were simulated to estimate the pharmacodynamic exposures for each patient. Pharmacodynamic thresholds for plazomicin, tigecycline, and meropenem were a total area under the curve to minimum inhibitory concentration ratio (AUC/MIC) ≥ 85, free (f) AUC/MIC ≥ 0.9, and free time above the MIC (fT > MIC) of ≥ 40%, respectively.ResultsFifteen plazomicin-treated patients were included (12 received tigecycline, 4 received meropenem, 1 received both). Microbiological response was observed in 13 (86.7%) and clinical efficacy was achieved in 11 (73.3%). Plazomicin achieved its pharmacodynamic target in all 15 patients. Meropenem fT > MIC was 0% in all 4 patients, and tigecycline fAUC/MIC was ≥ 0.9 in 9 (75%) patients. Overall, 6 (40%) of 15 patients had a tigecycline or meropenem exposure below the requisite thresholds. Microbiological response and clinical efficacy were observed in 100% (6/6) and 83.3% (5/6) of patients with low threshold attainment by tigecycline and meropenem dosing regimens, respectively.ConclusionsPlazomicin successfully achieved its requisite pharmacodynamic exposure, and these data suggest that optimization of tigecycline and meropenem therapy was not required for the combination to achieve microbiological response and clinical efficacy against serious CRE infections.Trial RegistrationClinicalTrials.gov number, NCT01970371.FundingAchaogen, Inc.Electronic supplementary materialThe online version of this article (10.1007/s40121-019-0251-4) contains supplementary material, which is available to authorized users.

Effects of sub-inhibitory concentrations of meropenem and tigecycline on the expression of genes regulating pili, efflux pumps and virulence factors involved in biofilm formation by Acinetobacter baumannii.

Background: Sub-minimal inhibitory concentrations of antibiotics have been indicated to affect the biofilm formation in pathogens of nosocomial infections. This study aimed to investigate the effects of meropenem and tigecycline at their sub-minimum inhibitory concentrations (MICs) on the biofilm formation capacity of Acinetobacter baumannii (A. baumannii), as well as the expression levels of genes involved in biofilm formation, quorum sensing, pili assembly and efflux pumps.Materials and methods: In this study, four non-clonal strains (AB10, AB13, AB32 and AB55), which were different from the aspects of antibiotic susceptibility and biofilm formation from each other were selected for the evaluation of antimicrobial susceptibility, biofilm inducibility at sub-MICs of meropenem and tigecycline and the gene expression levels (the abaI, abaR, bap, pgaA, csuE, bfmS, bfmR, ompA, adeB, adeJ and adeG genes).Result: A significant increase in the MICs of all antibiotics was demonstrated in the biofilm cells in each four strains. The biofilm formation was significantly decreased in all the representative strains exposed to tigecycline. However, the biofilm inducibility at sub-MICs of meropenem was dependent on strain genotype. In concordance with these results, Pearson correlation analysis indicated a positive significant correlation between the biofilm formation capacity and the mRNA levels of genes encoding efflux pumps except adeJ, the genes involved in biofilm formation, pili assembly and quorum sensing following exposure to meropenem and tigecycline at their sub-MICs.Conclusion: These results revealed valuable data into the correlation between the gene transcription levels and biofilm formation, as well as quorum sensing and their regulation at sub-MICs of meropenem and tigecycline.

Utility of the fluorogenic characters of benzofurazan for analysis of tigecycline using spectrometric technique; application to pharmacokinetic study, urine and pharmaceutical formulations.

Tigecycline (TIGE) is the newest tetracycline derivative antibiotic with low toxicity, it is used for management of infectious diseases caused by Gram-positive and Gram-negative bacteria. Hence, an efficient, selective and sensitive method was developed for analysis of TIGE in commercial formulations, human plasma and urine. The spectrofluorimetric technique based on the reaction of secondary amine moiety in TIGE with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in slightly alkaline medium producing a highly fluorescent product measured at 540nm (λex at 470nm) after heating for 15min at 75°C. The proposed strategy was upgraded and approved by ICH rules and bio-analytical validated using US-FDA recommendations. A linear relationship between fluorescence intensity and TIGE concentration was observed over the concentration range 40-500ngmL-1 with limit of quantification (LOQ) 21.09ngmL-1 and limit of detection (LOD) 6.96ngmL-1 .The ultra-affectability and high selectivity of the proposed strategy permits analysis of TIGE in dosage form, human plasma and urine samples with good recovery ranged from 97.23% to 98.72% and from 99.36% to 99.80% respectively, without any interfering from matrix components. Also, the developed strategy was used to examine the stability of TIGE in human plasma and applied for pharmacokinetic investigation of TIGE.

Different influences on mitochondrial function, oxidative stress and cytotoxicity of antibiotics on primary human neuron and cell lines

Although antibiotics are generally well tolerated, their toxic effects on the central nervous system have been gained attention. In this study, we systematically investigated the neuron toxicity of antibiotics from six different classes. We show that clinically relevant concentrations of metronidazole, tigecycline, azithromycin and clindamycin but not ampicillin or sulfamethoxazole induce apoptosis of human primary neuron cells and lines. Notably, tigecycline, azithromycin and clindamycin cause neuron cell oxidative damage whereas metronidazole has no effect on reactive oxygen species (ROS) production, suggesting that metronidazole induces neuron death via ROS-independent mechanism. Tigecycline, azithromycin and clindamycin induce mitochondrial dysfunctions via targeting different mitochondrial respiratory complexes, leading to mitochondrial membrane potential disruption and energy crisis. The deleterious effects of antibiotics are reversed by pretreatment of neuron cells with antioxidant. Our work highlights the different influences of antibiotics on mitochondrial dysfunction, oxidative damage and cytotoxicity in neuron cells. We also provide a strategy to prevent the neurotoxicity.

In-vitro activities of imipenem–colistin, imipenem–tigecycline, and tigecycline–colistin combinations against carbapenem-resistant Enterobacteriaceae*

The aim of the study is to determine in-vitro effects of imipenem–tigecycline, imipenem–colistin and tigecycline–colistin against carbapenem-resistant Enterobacteriaceae (CRE) isolates. A total of 25 CRE isolates were included to the study. The minimum inhibition concentrations of imipenem, colistin-sulphate and tigecycline were determined with broth dilution method. Synergistic effects of imipenem–tigecycline, imipenem–colistin and tigecycline–colistin were investigated by microdilution checkerboard technique. All of the isolates were resistant to imipenem, whereas 25% of the isolates were resistant to colistin and tigecycline. Imipenem–colistin, imipenem–tigecycline and tigecycline–colistin combinations were synergistic against 40% (10/25), 24% (6/25), and 36% (9/25) of the isolates, respectively. Antagonism was observed in 8% (2/25) of the isolates in tigecycline–colistin combination. Tigecycline–colistin was the most effective (70% synergy) combination in Klebsiella spp. strains; whereas imipenem–colistin was the most effective (75% synergy) combination in Escherichia coli strains. Synergistic effect was variable and strain-depended against CRE isolates that have been tested.

Antibacterial Efficacy and Discoloration Potential of Endodontic Topical Antibiotics

IntroductionThe optimal concentration for the use of endodontic topical antibiotics is not known. The aims of this study were to determine the minimum bactericidal concentrations (MBCs) and minimum inhibitory concentrations (MICs) of metronidazole, ciprofloxacin, minocycline, Augmentin (GlaxoSmithKline, Research Triangle Park, NC), and tigecycline against common endodontic pathogens and to evaluate ex vivo the antibacterial efficacy and discoloration effect of triple antibiotic paste (TAP), Augmentin, and tigecycline at different concentrations using a slow-release hydrogel scaffold. MethodsUsing the Epsilometer test method (Etest; bioMérieux USA, St Louis, MO), MICs and MBCs of selected antibiotics were determined against Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus intermedius, and Enterococcus faecalis. Biofilms of these bacterial species were then grown in extracted single-rooted teeth anaerobically for 3 weeks. Root canals were filled with TAP, Augmentin, and tigecycline at concentrations of 1 or 0.1 mg/mL in a degradable hydrogel scaffold or pure TAP at 1 g/mL for 7 days. Coronal discoloration was evaluated spectrophotometrically at 1, 2, and 3 weeks after dressing. ResultsMIC/MBC data showed significant efficacy of tigecycline, Augmentin, and minocycline compared with the other antibiotics (P < .05). Significant differences were found when comparing the log10 colony-forming units of all experimental groups (P < .05). TAP at 1 g/mL had no bacterial growth but caused the greatest discoloration. Hydrogel mixtures with TAP, Augmentin, or tigecycline at 1 mg/mL significantly reduced bacterial growth and the number of positive samples compared with those at 0.1 mg/mL (P < .05) with minimal discoloration. ConclusionsTAP, Augmentin, and tigecycline in a hydrogel at 1 mg/mL reduced bacterial growth significantly with minimal color change.

Comparison of Omadacycline and Tigecycline Pharmacokinetics in the Plasma, Epithelial Lining Fluid, and Alveolar Cells of Healthy Adult Subjects.

ABSTRACTThe steady-state concentrations of omadacycline and tigecycline in the plasma, epithelial lining fluid (ELF), and alveolar cells (AC) of 58 healthy adult subjects were obtained. Subjects were administered either omadacycline at 100 mg intravenously (i.v.) every 12 h for two doses followed by 100 mg i.v. every 24 h for three doses or tigecycline at an initial dose of 100 mg i.v. followed by 50 mg i.v. every 12 h for six doses. A bronchoscopy and bronchoalveolar lavage were performed once in each subject following the start of the fifth dose of omadacycline at 0.5, 1, 2, 4, 8, 12, or 24 h and after the start of the seventh dose of tigecycline at 2, 4, 6, or 12 h. The value of the area under the concentration-time curve (AUC) from time zero to 24 h postdosing (AUC0–24) (based on mean concentrations) in ELF and the ratio of the ELF to total plasma omadacycline concentration based on AUC0–24 values were 17.23 mg · h/liter and 1.47, respectively. The AUC0–24 value in AC was 302.46 mg · h/liter, and the ratio of the AC to total plasma omadacycline concentration was 25.8. In comparison, the values of the AUC from time zero to 12 h postdosing (AUC0–12) based on the mean concentrations of tigecycline in ELF and AC were 3.16 and 38.50 mg · h/liter, respectively. The ratio of the ELF and AC to total plasma concentrations of tigecycline based on AUC0–12 values were 1.71 and 20.8, respectively. The pharmacokinetic advantages of higher and sustained concentrations of omadacycline compared to those of tigecycline in plasma, ELF, and AC suggest that omadacycline is a promising antibacterial agent for the treatment of lower respiratory tract bacterial infections caused by susceptible pathogens.

Pharmacokinetics of tigecycline in turkeys following different routes of administration.

The aim of this research had been to determine the pharmacokinetics of tigecycline (TIG) in turkey after intravenous (i.v.), intramuscular (i.m.), subcutaneous (s.c.), and oral (p.o.) administration at a dose of 10mg/kg. TIG concentrations in plasma were determined using high-performance liquid chromatography with tandem mass spectrometry. Mean concentrations of TIG in turkey plasma in the i.v. group were significantly higher than concentrations of this drug obtained after using the other administration routes. No significant differences were demonstrated in respect to the concentrations achieved after i.m. and s.c. administration. The bioavailability of TIG after i.m., s.c., and p.o. administration was 32.59±5.99%, 34.91±9.62%, and 0.97±0.57%, respectively. Values of half-life in the elimination phase were 23.49±6.51hr, 25.42±4.42hr, and 26.62±5.19hr in i.v., i.m., and s.c. groups, respectively, values of mean residence time were 7.92±1.41hr, 19.62±2.82hr, and 17.55±2.59hr in i.v., i.m., and s.c. groups, respectively, whereas the volume of distribution was 14.85±5.71L/kg, 14.68±2.56L/kg, and 15.37±3.00L/kg in i.v., i.m., and s.c. groups, respectively. Because TIG is not absorbed from the gastrointestinal tract in turkeys to a clinically significant degree, this drug given p.o. could find application in commercial turkey farms only to treat gastrointestinal tract infections.

Population Pharmacokinetics of Tigecycline in Critically Ill Patients with Severe Infections.

We sought to describe the population pharmacokinetics of tigecycline in critically ill patients and to determine optimized dosing regimens of tigecycline for different bacterial infections. This prospective study included 10 critically ill patients given a standard dose of tigecycline. Blood samples were collected during one dosing interval and were analyzed using validated chromatography. Population pharmacokinetics and Monte Carlo dosing simulations were undertaken using Pmetrics. Three target exposures, expressed as ratios of the 24-h area under the curve to MICs (AUC0-24/MIC), were evaluated (≥17.9 for skin infections, ≥6.96 for intra-abdominal infections, ≥4.5 for hospital-acquired pneumonia). The median age, total body weight, and body mass index (BMI) were 67 years, 69.1 kg, and 24.7 kg/m2, respectively. A two-compartment linear model best described the time course of tigecycline concentrations. The parameter estimates (expressed as means ± standard deviations [SD]) from the final model were as follows: clearance (CL), 7.50 ± 1.11 liters/h; volume in the central compartment, 72.50 ± 21.18 liters; rate constant for tigecycline distribution from the central to the peripheral compartment, 0.31 ± 0.16 h-1; and rate constant for tigecycline distribution from the peripheral to the central compartment, 0.29 ± 0.30 h-1 A larger BMI was associated with increased CL of tigecycline. Licensed doses were found to be sufficient for Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, and methicillin-resistant Staphylococcus aureus for an AUC0-24/MIC target of 4.5 or 6.96. For a therapeutic target of 17.9, an increased tigecycline dose is required, especially for patients with higher BMI. The dosing requirements of tigecycline differ with the indication, with pathogen susceptibility, and potentially with patient BMI.

Ocular penetration of topically applied 1% tigecycline in a rabbit model.

To evaluate ocular penetration of topically applied 1% tigecycline. Forty-two New Zealand White rabbits were divided into 3 groups. A 50 µL drop of 1% tigecycline was administered in group 1. In groups 2 and 3, the drop was administered every 15min for 60min (keratitis protocol). Aqueous humor samples in groups 1 and 2 were collected under general anesthesia at 15, 30, 45, 60, 120, and 180min after the last drop. All animals in group 3 were euthanatized. Cornea, vitreous and blood samples were collected 60 and 120min after the last drop. Tigecycline concentrations were measured using high performance liquid chromatography-mass spectrometry (LC-MS/MS). The peak aqueous humor tigecycline concentration [mean 0.73±0.14 mg/L (SD) and 2.41±0.14 mg/L, respectively] occurred 45min after topical drug application in groups 1 and 2. Group 3 mean values in the cornea, and vitreous, were 3.27±0.50 µg/g, and 0.17±0.10 mg/L at 60min and 3.17±0.77 µg/g and 0.20±0.07 mg/L at 120min, respectively. Tigecycline serum concentrations were negligible. Tigecycline levels in the aqueous humor in groups 1 and 2, and in the cornea in group 3 exceeded the minimum inhibitory concentrations of most gram-positive organisms that cause bacterial keratitis and endophthalmitis.

Effect of subinhibitory concentrations of tigecycline and ciprofloxacin on the expression of biofilm-associated genes and biofilm structure of Staphylococcus epidermidis.

Staphylococcus epidermidis is a leading cause of foreign body-associated infections. This is related to the bacterium's ability to form biofilms on synthetic materials. Bacteria within a biofilm may be exposed to subinhibitory concentrations (sub-MICs) of antibiotics because of an agent's limited penetration into the biofilm core. Here, we investigated the effect of sub-MICs of tigecycline and ciprofloxacin on the expression of biofilm-associated genes, i.e. icaA, altE and sigB, and the biofilm structure of five clinical isolates of S. epidermidis. For most tested isolates, the expression of these genes increased after exposure to 0.25 MIC and 0.5 MIC tigecycline. A slight decrease in icaAmRNA levels was observed only in two isolates in the presence of 0.25 MIC tigecycline. The effect of ciprofloxacin exposure was isolate-dependent. At 0.5 MIC, ciprofloxacin induced an increase of sigB and icaAmRNA levels in three of the five tested isolates. At the same time, expression of the altE gene increased in all isolates (from 1.3-fold to 42-fold, depending on the strain). Confocal laser scanning microscopy analysis indicated that sub-MIC ciprofloxacin decreased biofilm formation, whereas tigecycline stimulated this process. Our data suggest that sub-MIC tigecycline may have bearing on the outcome of infections.

Pharmacodynamics of tigecycline alone and in combination with colistin against clinical isolates of multidrug-resistant Acinetobacter baumannii in an in vitro pharmacodynamic model.

The objectives of this study, which were based on the hypothesis of mutant prevention concentration (MPC), were to compare tigecycline and colistin monotherapy and combination therapy against multidrug-resistant Acinetobacter baumannii (MDR-AB) and to identify changes in the susceptibility of the organism using an in vitro pharmacodynamic model. Human free-drug concentration profiles of colistin and tigecycline used alone and in combination were simulated against four clinical MDR-AB isolates over 24 h. Pharmacodynamic activity was measured as log10 CFU/mL and as the area under the bactericidal curve (AUBC). The minimum inhibitory concentration (MIC) for all isolates was determined in triplicate by the broth microdilution method. All isolates grew to control levels in the tigecycline and colistin monotherapy conditions, and the combination of colistin plus tigecycline 100 mg every 12 h (q12h) or 50 mg q12h achieved a greater reduction in bacterial density than colistin alone (-2.65 ± 1.73 or -2.09 ± 1.47 vs. 0.98 ± 0.64 log10 CFU/mL; P <0.01). Likewise, both combinations significantly reduced the AUBC compared with that achieved using colistin alone (106.9 ± 24.5 or 117.7 ± 23.5 vs. 168.1 ± 14.2 log10 CFU⋅h/mL; P <0.05). When tigecycline or colistin monotherapy concentrations were below MPC, tigecycline MICs increased 4-32-fold and colistin MICs increased >16-fold. No loss in susceptibility to tigecycline was found with combination therapy. A combination of tigecycline (high dose) and colistin may be an effective therapy to synergistically prevent the emergence of resistance during treatment of MDR-AB (tigecycline MIC < 2 mg/L) infections.

Comparison of minimal inhibitory concentrations (MICs) of tigecycline in 2011 and 2015 years against multidrug-resistance acinetobacter baumannii strains

Acinetobacter baumannii is an opportunistic pathogen that causing nosocomial infections. The microorganisms can be developed resistance to antimicrobials very quickly by different mechanisms. Infections defend by MDR strains, caused require long-term hospitalization high morbidity and mortality. Tigecycline is one of the few antimicrobials which have activity against MDR A. baumannii strains. In Meta-analyzes and clinical study reported that a rising risk of mortality in tigecycline-treated patients due to an increase in resistance of these strains. In our study, we aimed to compare the MICs values of tigecycline and determine the resistance rates of tigecycline against to MDR A. baumannii strains in 2011 and 2015. Tigecycline MIC values of 200 A. baumannii isolates were determined by the broth microdilution method. Final concentrations were prepared as 16μg/ml to 0.06μg/ml. Tigecycline MICs breakpoint values comments on breakpoint values of Enterobacteriaceae that have been proposed by FDA. In 2011, tigecycline values of MIC50 and MIC 90 found 0.5μg/ml, 1μg/ml, respectively against to MDR A. baumannii strains. In 2015, tigecycline values of MIC50 and MIC90 found 1μg/ml, 4μg/ml, respectively. In 2011 the 4%, while in 2015, 16% of A. baumannii strains� tigecycline MICs were determined over 2μg/ml. and that this was interpreted as the resistance. Tigecycline can be still use as an alternative antimicrobial for treatment of A. baumannii strains, especially in our hospitals� intensive care units. However, to avoid the development of resistance against tigecycline, we should prevent the irrational use of antibiotics. The implementation of treatment should be based on antimicrobial susceptibility test results.

A Novel Microfluidic Assay for Rapid Phenotypic Antibiotic Susceptibility Testing of Bacteria Detected in Clinical Blood Cultures.

BackgroundAppropriate antibiotic therapy is critical in the management of severe sepsis and septic shock to reduce mortality, morbidity and health costs. New methods for rapid antibiotic susceptibility testing are needed because of increasing resistance rates to standard treatment.AimsThe purpose of this study was to evaluate the performance of a novel microfluidic method and the potential to directly apply this method on positive blood cultures.MethodsMinimum inhibitory concentrations (MICs) of ciprofloxacin, ceftazidime, tigecycline and/or vancomycin for Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus were determined using a linear antibiotic concentration gradient in a microfluidic assay. Bacterial growth along the antibiotic gradient was monitored using automated time-lapse photomicrography and growth inhibition was quantified by measuring greyscale intensity changes in the images. In addition to pure culture MICs, vancomycin MICs were determined for S. aureus from spiked and clinical blood cultures following a short centrifugation step. The MICs were compared with those obtained with the Etest and for S. aureus and vancomycin also with macrodilution.ResultsThe MICs obtained with the microfluidic assay showed good agreement internally as well as with the Etest and macrodilution assays, although some minor differences were noted between the methods. The time to possible readout was within the range of 2 to 5 h.ConclusionsThe examined microfluidic assay has the potential to provide rapid and accurate MICs using samples from positive clinical blood cultures and will now be tested using other bacterial species and antibiotics.

Biofilm Formation Restrained by Subinhibitory Concentrations of Tigecyclin in Acinetobacter baumannii Is Associated with Downregulation of Efflux Pumps

Background: Tigecycline, one of the few therapeutic options against multidrug-resistant Acinetobacter baumannii, reaches subinhibitory serum concentrations only with cautious clinical dosing and pharmacokinetics. Subinhibitory concentrations of tigecycline might induce an A. baumannii biofilm. Methods: Biofilm formation was assessed via the crystal violet staining method. We further analyzed the main biofilm components with NaIO₄, proteinase K, and DNase. Real-time RT-PCR was applied for quantitative detection of biofilm potential-associated genes. Results: In this study, A. baumannii proved to be a strong biofilm producer, and we found that proteins and extracellular DNA are crucial components of the A. baumannii biofilm. Quantitative real-time RT-PCR revealed positive correlations between biofilm formation restrained by subinhibitory concentrations of tigecycline and the expression of biofilm potential-associated genes, especially the AdeFGH efflux pump gene. Conclusion: Our results suggest that downregulation of efflux pumps, especially the AdeFGH efflux pump, is probably responsible for the decline in biofilm formation in A. baumannii treated with subinhibitory concentrations of tigecyclin.