Colistin-resistant Acinetobacter Baumannii Research Articles

Discovery of new antimicrobial thiophene derivatives with activity against drug-resistant Gram negative-bacteria.

Our aim is to identify new small molecules with antimicrobial potential, especially against colistin-resistant (Col-R) Acinetobacter baumannii and Escherichia coli. After initial hits identification by fingerprint similarity, MIC of 24 heterocyclic derivatives for A. baumannii and E. coli reference strains, and bactericidal activity of selected thiophenes against Col-R strains were determined. We analyzed changes in bacterial membrane permeability and the OMPs profile. Additionally, we determined bacterial adherence to host cells and performed molecular docking studies to assess their binding to bacterial targets. The compounds' MICs ranged from 4 to >64mg/L. Thiophene derivatives 4, 5 and 8 exhibited MIC50 values between 16 and 32mg/L for Col-R A. baumannii and 8 and 32mg/L for Col-R E. coli. The time-kill curve assay demonstrated that thiophenes 4 and 8 had bactericidal effects against Col-R A. baumannii and E. coli. Furthermore, treatment with them resulted in increased membrane permeabilization and reduced adherence of these isolates to host cells. Finally, the docking studies showed a stronger binding affinity to CarO1 and Omp33 of A. baumannii and OmpW and OmpC of E. coli. These findings indicate that thiophene derivatives possess antibacterial activity against Col-R A. baumannii and E. coli, suggesting that they may enhance the repertoire of drug treatments against bacteria.

Isolation, Identification, and Antibiogram of Colistin-resistant Acinetobacter baumannii from Rivers in and around Kathmandu Valley

Background: Acinetobacter baumannii, an opportunistic Gram-negative pathogen, poses an escalating threat in clinical settings due to the rise of multidrug-resistant infections. Despite its clinical significance, there exists a considerable gap in understanding its environmental dissemination.
 Aims and Objectives: The primary objective is to examine the distribution of A. baumannii and its antibiotic resistance in river ecosystems. Specifically, we aim to identify strains resistant to Colistin, a last-resort antibiotic, and elucidate the susceptibility patterns to other antibiotics.
 Materials and Methods: Water samples from 10 rivers were collected and subjected to analysis using Leeds Acinetobacter Agar Base and a series of biochemical tests. Antibiotic susceptibility testing, focusing on Colistin resistance, was performed using standard procedures.
 Results: Out of the 284 isolated strains, 14 (4.9%) exhibited resistance to Colistin, while demonstrating varying susceptibility patterns to other antibiotics. Notably, Gentamycin showed effectiveness against resistant strains (14.28%), while Ceftazidime resistance was complete. Colistin-sensitive strains displayed high susceptibility to Ciprofloxacin (84.44%) and lower susceptibility to Chloramphenicol (53.33%). Carbapenem susceptibility was observed across all isolates.
 Conclusion: The study underscores a concerning environmental presence of multidrug-resistant A. baumannii in rivers around Kathmandu Valley, with Sundarijal being the exception. The findings emphasize the necessity of scrutinizing environmental reservoirs for pathogen spread, advocating for heightened awareness of potential health implications beyond clinical settings. Urgent attention is needed to comprehend and counteract the emergence and dissemination of antibiotic resistance, necessitating comprehensive strategies and continued surveillance

Molecular mechanisms of colistin resistance mediated by pmrCAB genes in Acinetobacter baumannii isolated from patients hospitalized in Isfahan medical centers

Abstract Introduction In recent years, colistin-resistant Acinetobacter baumannii (A. baumannii) has been found all over the world. In this current study, the main purpose was to examine the occurrence of extensively drug resistant (XDR), resistance to colistin and characterization and mutations in pmrCAB genes among A. baumannii obtained from inpatients. Materials and Methods A total of 108 clinical isolates of A. baumannii were collected from several hospitals located in Isfahan, Iran. The Kirby-Bauer assay was performed to assess the antimicrobial resistance. The Phoenix automated system was utilized to determine the minimum inhibitory concentration (MIC) of colistin for each of the bacterial isolates. Polymerase chain reaction was used to screen for pmrCAB genes that mediate colistin resistance, and sequencing was used to determine the amplicon’s nucleotide sequence. Results The results revealed that all A. baumannii isolates (100%) were resistant to piperacillin tazobactam, meropenem and ciprofloxacin. All isolates were classified as XDR, with seven isolates being pan-drug resistant (PDR). Colistin resistance (CoR) was found in 6.48% (7/108) of studied isolates, all of which were positive for pmrCAB genes. The sequencing results showed a substitution in pmrB and two isolates showed a substitution in pmrC. Conclusions In conclusion, this study is the initial report of the existence and mutations of pmrB and C genes in the clinical isolate of A. baumannii in our region. This outcome highlights the necessity to explore additional mutations in the PMR operon of A. baumannii in forthcoming studies. Moreover, our results highlight the high occurrence of XDR-A. baumannii strain in Isfahan, Iran.

The efficacy of mesenchymal stem cell treatment and colistin-fosfomycin combination on colistin-resistant Acinetobacter baumannii sepsis model.

This study examines the role of mesenchymal stem cells (MSCs) in an experimental sepsis model developed with colistin-resistant Acinetobacter baumannii (CRAB). BALB-c mice were divided into treatment groups (MSC, MSC + colistin (C)-fosfomycin (F), and C-F and control groups (positive and negative)). CRAB was administered to mice through intraperitoneal injection. Three hours later, C, F, and MSC were given intraperitoneally to the treatment groups. Colistin administration was repeated every 12 h, F administration was done every 4 h, and the second dose of MSC was administered after 48 h. Mice were sacrificed at 24 and 72 h. The bacterial load was determined as colony-forming units per gram (cfu/g). Histopathological examination was conducted on the left lung, liver, and both kidneys. IL-6 and C-reactive protein (CRP) levels in mouse sera were determined by enzyme-linked immunosorbent assay. Among the treatment groups, the C-F group had the lowest colony count in the lung (1.24 ± 1.66 cfu/g) and liver (1.03 ± 1.08 cfu/g). The highest bacterial clearance was observed at 72 h compared to 24 h in the MSC-treated groups (p = 0.008). The MSC + C-F group showed the lowest histopathological score in the liver and kidney (p = 0.009). In the negative control group, the IL-6 level at the 24th hour was the lowest (p < 0.001). Among the treatment groups, the CRP level was the lowest in the MSC + C-F group at 24 and 72 h. In a CRAB sepsis model, adding MSCs to a colistin-fosfomycin treatment may be beneficial in terms of reducing bacterial loads and preventing histopathological damage.

Emergence of colistin resistant Acinetobacter baumannii clonal complex 2 (CC2) among hospitalized patients in Iran.

Acinetobacter baumannii is a major causative agent of serious nosocomial infections. This study was carried out to investigate the molecular characterization of colistin resistant isolates of A. baumannii from hospitalized patients, based on multilocus sequence typing (MLST). A cross-sectional study was conducted to collect A. baumannii from clinical samples in Isfahan from 2021 to 2022. Isolates were identified as A. baumannii using biochemical tests and PCR of blaOXA-51. Antibiotic susceptibility testing was carried out using the Kirby-Bauer method and minimum inhibitory concentration (MIC) values were determined for colistin. Additionally, MLST was performed according to the Pasteur scheme to assess the relationship between colistin resistant A. baumannii. A total of 70 non-repetitive A.baumannii isolates were obtained from different clinical samples. MIC results showed that seven A.baumannii isolates were resistant to colistin. The antibiotic susceptibility pattern revealed that all seven colistin resistant strains were resistant to all tested antibiotics. Based on MLST analysis, the colistin resistant isolates were assigned to five unique STs namely, ST2 (3; 42.9%) followed by ST78 (1;14.3%), ST1077 (1; 14.3%), ST415 (1; 14.3%) and ST391 (1; 14.3%). Among them ST2, ST391 and ST415 belong to clonal complex 2. Colistin resistant A. baumannii ST2 is the main circulating clone in clinical settings in Iran, but additionally ST415, ST391, and ST1077 are found for the first time in our country. Intensive control procedures and strict adherence to surveillance programs are recommended to decrease the spread of carbapenem and colistin resistant A. baumannii strain.

Evaluation of potential synergistic activity of antimicrobial combinations against Colistin resistant Acinetobacter baumannii

Evaluation of potential synergistic activity of antimicrobial combinations against Colistin resistant Acinetobacter baumannii

Risk factors associated with colistin-resistant Acinetobacter baumannii infection

Acinetobacter baumannii is an important cause of healthcare-associated infections and is resistant to almost all antimicrobial agents, with strains recently reported to be resistant to colsitin. In this study, we aimed to identify the risk factors associated with colistin-resistant A. baumannii infections by comparing colistin-resistant and -susceptible A. baumannii isolates. We retrospectively reviewed the medical records of 51 and 100 cases in which colistinresistant and -susceptible A. baumannii were isolated, respectively. Univariate analysis showed that compared with patients with colistin-sensitive infections, patients with colistinresistant A. baumanni infections had a combined pulmonary disease (P = 0.017), were admitted to intensive care unit (P = 0.020), and had prior mechanical ventilation (P = 0.003), tracheostomy (P = 0.043), percutaneous drainage (P = 0.070), hemodialysis (P = 0.002); use of colistin (P = 0.000), carbapenem (P = 0.000), and teicoplanin (P = 0.004); and co-infection (P = 0.035). Multivariate analysis indicated that eight variables were related to the likelihood of colistin-resistant A. baumanni infections: use of teicoplanin (Odds ratio [OR]: 3.140, 95% confidence interval [CI]: 0.529–18.650), prior hemodialysis (OR: 2.722, 95% CI: 0.851–8.709), combined pulmonary disease (OR: 2.286, 95% CI: 0.998–5.283), prior use of carbapenem (OR: 0.199, 95% CI: 0.863–5.603), co-infection (OR: 1.706, 95% CI: 0.746–3.898), prior mechanical ventilation (OR: 1.614, 95% CI, 0.684–3.809), intensive care unit admission (OR: 1.387, 95% CI: 0.560–3.435), and prior tracheostomy (OR: 1.102, 95% CI: 0.344–3.527); however, no statistical differences were observed. Although colistin use could not be proven in multivariate analysis, the possibility of being a risk factor cannot be ruled out.

Quercetin: Synergistic Interaction with Antibiotics against Colistin-Resistant Acinetobacter baumannii.

Infections caused by resistant strains of Acinetobacter baumannii are now a global problem that requires the immediate development of new antimicrobial drugs. Combination therapy is one of the strategies used to solve this problem. Based on this information, the purpose of this study was to determine whether quercetin (QUE), in combination with three antibiotics, is effective against colistin-resistant A. baumannii strains (ColR-Ab). The effects of the combination of QUE with colistin (COL), amikacin (AMK), and meropenem (MEM) were evaluated according to the checkerboard synergy test. The combinations of QUE + COL and QUE + AMK showed synergistic activity on ColR-Ab strains with FICI values in the range of 0.1875-0.5 and 0.1875-0.2825, respectively. A 4- to 16-fold decrease in COL MIC and a 16- to 64-fold decrease in AMK MIC values were detected. Synergistic activity was confirmed by the time-kill test, and these combinations were found to be bactericidal at the end of 24 h. According to spectrophotometric measurements, the combinations of QUE + COL and QUE + AMK induced membrane damage, leading to the leakage of nucleic acids. Cell lysis and cell death were confirmed with SEM observations. The detected synergy offers an opportunity for the future development of treatment strategies for potential infections caused by ColR-Ab strains.

In Vitro Synergistic Activity of Antimicrobial Combinations against Carbapenem- and Colistin-Resistant Acinetobacter baumannii and Klebsiella pneumoniae.

Polymyxins are commonly used as the last resort for the treatment of MDR Acinetobacter baumannii and Klebsiella pneumoniae nosocomial infections; however, apart from the already known toxicity issues, resistance to these agents is emerging. In the present study, we assessed the in vitro synergistic activity of antimicrobial combinations against carbapenem-resistant and colistin-resistant A. baumannii and K. pneumoniae in an effort to provide more options for their treatment. Two hundred A. baumannii and one hundred and six K. pneumoniae single clinical isolates with resistance to carbapenems and colistin, recovered between 1 January 2021 and 31 July 2022,were included. A. baumannii were tested by the MIC test strip fixed-ratio method for combinations of colistin with either meropenem or rifampicin or daptomycin. K. pneumoniae were tested for the combinations of colistin with meropenem and ceftazidime/avibactam with aztreonam. Synergy was observed at: 98.99% for colistin and meropenem against A. baumannii; 91.52% for colistin and rifampicin; and 100% for colistin and daptomycin. Synergy was also observed at: 73.56% for colistin and meropenem against K. pneumoniae and; and 93% for ceftazidime/avibactam with aztreonam. The tested antimicrobial combinations presented high synergy rates, rendering them valuable options against A. baumannii and K. pneumoniae infections.

Comparative genomics and molecular epidemiology of colistin-resistant Acinetobacter baumannii

This study aimed to investigate the prevalence and resistance mechanisms of colistin-resistant Acinetobacter baumannii (ColRAB) isolates in Serbia, assess their genetic relatedness to other circulating A. baumannii isolates in the neighbouring European countries, and analyse the global genomic epidemiology of ColRAB isolates. A total of 784 isolates of A. baumannii were recovered from hospitalised patients in Serbia between 2018 and 2021. The antimicrobial susceptibility testing was performed using disk diffusion and broth microdilution. All ColRAB isolates were subjected to DNA isolation and whole-genome sequencing (WGS). Overall, 3.94 % (n = 30) isolates were confirmed as ColRAB. Results of mutational and transcriptional analysis of genes associated with colistin resistance indicate the central role of the two-component regulating system, PmrAB, and increased expression of the pmrC gene in ColRAB. Most of the isolates (n = 29, 96.6 %) belonged to international clone II, with the most common sequence type being STPas2 (n = 23, 76.6 %). Based on the WGS analysis, ColRAB isolates belonging to the same ST isolated in various countries were grouped into the same clusters, indicating the global dissemination of several high-risk clonal lineages. Phylogenomic analysis of ColRAB isolates, together with all previously published A. baumannii genomes from South-Eastern European countries, showed that colistin resistance arose independently in several clonal lineages. Comparative genomic analysis revealed multiple genes with various roles (transcriptional regulation, transmembrane transport, outer membrane assembly, etc.), which might be associated with colistin resistance in A. baumannii. The obtained findings serve as the basis for further studies, contributing to a better understanding of colistin resistance mechanisms in A. baumannii.

C-Locked Analogs of the Antimicrobial Peptide BP214.

BP214 is an all-D antimicrobial peptide amide, kklfkkilryl, which shows an excellent activity against colistin-resistant Acinetobacter baumannii and a low hemolytic activity. The aim of the present work was to investigate how C-terminus-to-side chain macrocyclization and fatty acid modification affect the antimicrobial and hemolytic activity of this peptide. In total, 18 analogs of BP214 were synthesized using a combination of Fmoc-based solid-phase peptide synthesis and the submonomer approach. Cyclization was achieved by reacting the ε-amino group of a C-terminal lysine residue with a bromoacetylgroup attached to the Nα amino group of the N-terminal amino acid, generating a secondary amine at which the exocyclic lipopeptide tail was assembled. Three different ring sizes (i.e., 3–5 amino acid residues) of C-locked analogs combined with fatty acids of different lengths (i.e., C10–C14) were investigated. The antimicrobial activity of the analogs was tested against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa. The most promising compound was analog 13 (MIC = 4 µg/mL (2.4 µM) against E. coli and 36% hemolysis of red blood cells at 150 µM). In a time-kill assay, this peptide showed a significant, concentration-dependent reduction in viable E. coli cells comparable to that seen for colistin.

Synergistic activities of colistin combined with other antimicrobial agents against colistin-resistant Acinetobacter baumannii clinical isolates.

Emerging resistance to colistin in Acinetobacter baumannii clinical strains is concerning because of the limited therapeutic choices for these important clinical pathogens. We studied the in vitro activities of different colistin-based antimicrobial agent combinations against colistin-resistant Acinetobacter baumannii. Fourteen clinical isolates of colistin-resistant Acinetobacter baumannii were obtained between 2015 and 2016. To identify colistin-based combinations with synergistic activities, multiple two antimicrobial combinations based on 8 commercially available drugs were evaluated by the checkerboard method. The most effective colistin-based combinations were vancomycin, aztreonam, ceftazidime and imipenem which showed synergistic activities against all examined strains. Colistin-rifampin showed synergy against four strains. Colistin-tigecycline and colistin-amikacin mostly showed indifferent results. By using the checkerboard tests, we were able to find the most promising colistin-based combinations that may provide more therapeutic options against colistin-resistant Acinetobacter baumannii.

Overcoming addition of phosphoethanolamine to lipid A mediated colistin resistance in Acinetobacter baumannii clinical isolates with colistin–sulbactam combination therapy

Overcoming colistin-resistant Acinetobacter baumannii (CoR-AB) has become a major concern due to the lack of effective antibiotics. This study aimed to explore the prevalence of CoR-AB clinical isolates in Thailand, their mechanisms of resistance, and test the efficacy of colistin plus sulbactam against CoR-AB isolates. The colistin resistance rate among carbapenem-resistant A. baumannii was 15.14%. The mcr gene or its variants were not detected in CoR-AB isolates by PCR screening. The lipid A mass spectra of CoR-AB isolates showed the additional [M–H]− ion peak at m/z = 2034 that correlated to the phosphoethanolamine (pEtN) addition to lipid A (N = 27/30). The important amino acid substitutions were found at position S14P, A138T, A227V in PmrB that are associated with overexpression of the pEtN transferase (PmrC) and contributed the pEtN addition. The lipopolysacccharide production genes (lpxACD) were not related to lipid A mass spectra. A colistin plus sulbactam combination exhibited the synergy rate at 86.7% against CoR-AB isolates compare to sulbactam (85.89% resistance) or colistin (15.14% resistance) alone. The excellent synergistic activity of colistin plus sulbactam combination has the potential for the treatment of CoR-AB infections.

MICROBIOLOGICAL ISOLATES AND IT’S RESISTOTYPE FROM CLINIC OF VASCULAR SURGERY FOR THE FIRST QUATER OF 2022

One of the most common complications of surgical exposures is the surgical site infection (SSI). Although it varies between different surgical profiles, it can reach up to one third of all complications. In vascular surgery patients ischemic ulcers are very common, as well as factors, compromising the immune system such as diabetes, chronic kidney disease etc. One of the main surgical exposures in vascular surgery is inguinal, providing access to the femoral artery and its bifurcation. Although it allows a wide range of reconstructions, implanting different types of prosthetic materials, stents and providing anastomosis site, it contains lymph nodes, which can contaminate the reconstruction and cause SSI with severe consequences. Patients, prone to SSI due to concomitant diseases, are threatened by sepsis, limb loss and even death, which makes prevention of those type of complications essential. Aim: To investigate etiological spectrum of microbiological isolates and their resistance against most common antimicrobials among vascular surgery patients. Materials and methods: The study is retrospective, conducted in the period 01 January 2022 – 31 March 2022. All of the samples were obtained from patients of Clinic of Vascular Surgery. After isolation of pure culture from the samples, the strains were identified by MALDI TOF MS and Vitec – 2 Compact. Antibiotic resistance was determined with Bauer-Kirby disk diffusion method. Results: From all 419 of the patients, hospitalized in the Clinic of Vascular surgery for this period, 28 isolates from 26 (6,21%) patients were obtained, of which Gram-negative were 19 (67,86%) and Gram-positive - 9 (32,14%). From Gram-negative - enterobacteria – 14 (73,68%), and non-fermenting gram-negative bacteria (NFGNB) were 5 (26,32%). Only 3 (21,43%) from all enterobacteria were extended spectrum beta-lactamases producing strains (ESBLs). No strains, resistant to carbapenems (RCP) were isolated. Five (55,55%) of the Gram-positive isolates were Staphylococcus aureus, 4 (80%) of which were methicillin resistant Staphylococcus aureus (MRSA). Two of the Gram-positive species isolated were Enterococcus faecalis, of which 1 with a high-level aminoglycoside resistance (HLAR). No Vancomycin resistant enterococci (VRE) were discovered. There were no colistin-resistant Acinetobacter baumannii and Pseudomonas aeruginosa strains. Conclusion: From all 28 isolates 8 (28,57%) were with acquired types of antimicrobial resistance. With almost one third of the isolates that are problematic in terms of antibiotic susceptibility, treatment of those patients can be challenging. Prevention of in hospital contamination with polyresistant strains, associated with medical care it is crucial for reducing the number of severe complications, decreasing of hospital stay and cost for treatment.

Whole-Genome Sequencing of a Colistin-Resistant Acinetobacter baumannii Strain Isolated at a Tertiary Health Facility in Pretoria, South Africa.

Background: Acinetobacter baumannii’s (A. baumannii) growing resistance to all available antibiotics is of concern. The study describes a colistin-resistant A. baumannii isolated at a clinical facility from a tracheal aspirate sample. Furthermore, it determines the isolates’ niche establishment ability within the tertiary health facility. Methods: An antimicrobial susceptibility test, conventional PCR, quantitative real-time PCR, phenotypic evaluation of the efflux pump, and whole-genome sequencing and analysis were performed on the isolate. Results: The antimicrobial susceptibility pattern revealed a resistance to piperacillin/tazobactam, ceftazidime, cefepime, cefotaxime/ceftriaxone, imipenem, meropenem, gentamycin, ciprofloxacin, trimethoprim/sulfamethoxazole, tigecycline, and colistin. A broth microdilution test confirmed the colistin resistance. Conventional PCR and quantitative real-time PCR investigations revealed the presence of adeB, adeR, and adeS, while mcr-1 was not detected. A MIC of 0.38 µg/mL and 0.25 µg/mL was recorded before and after exposure to an AdeABC efflux pump inhibitor. The whole-genome sequence analysis of antimicrobial resistance-associated genes detected beta-lactam: blaOXA-66; blaOXA-23; blaADC-25; blaADC-73; blaA1; blaA2, and blaMBL; aminoglycoside: aph(6)-Id; aph(3″)-Ib; ant(3″)-IIa and armA) and a colistin resistance-associated gene lpsB. The whole-genome sequence virulence analysis revealed a biofilm formation system and cell–cell adhesion-associated genes: bap, bfmR, bfmS, csuA, csuA/B, csuB, csuC, csuD, csuE, pgaA, pgaB, pgaC, and pgaD; and quorum sensing-associated genes: abaI and abaR and iron acquisition system associated genes: barA, barB, basA, basB, basC, basD, basF, basG, basH, basI, basJ, bauA, bauB, bauC, bauD, bauE, bauF, and entE. A sequence type classification based on the Pasteur scheme revealed that the isolate belongs to sequence type ST2. Conclusions: The mosaic of the virulence factors coupled with the resistance-associated genes and the phenotypic resistance profile highlights the risk that this strain is at this South African tertiary health facility.

Whole-genome sequencing for the characterization of resistance mechanisms and epidemiology of colistin-resistant Acinetobacter baumannii.

BackgroundMultidrug-resistant Acinetobacter baumannii is an important causal pathogen of healthcare-associated infections, and colistin-resistant strains have recently emerged owing to the increased use of colistin. Using next-generation sequencing (NGS), a single whole-genome sequencing (WGS) protocol can identify and type pathogens, analyze genetic relationships among different pathogens, predict pathogenic transmissions, and detect antibiotic resistance genes. However, only a few studies have applied NGS in studying the resistance mechanism and epidemiology of colistin-resistant A. baumannii. This study aimed to elucidate the resistance mechanism of colistin-resistant A. baumannii and analyze its molecular epidemiology through WGS.Materials and methodsThe subjects in this study were patients who visited a university hospital between 2014 and 2018. Thirty colistin-resistant strains with high minimum inhibitory concentrations were selected from various patient samples, and WGS was performed. Comparative genomic analysis was performed for the 27 colistin-resistant A. baumannii strains using a colistin-susceptible strain as the reference genome.ResultsThe WGS analysis found no mutation for lpxA, lpxC, lpx D, pmrA, pmrB, and mcr1, the genes known to be associated with colistin resistance. Fifty-seven coding sequences (CDS) showed differences; they included 13 CDS with known names and functions that contained 21 genes. From the whole-genome multi-locus sequence typing (wgMLST) and single nucleotide polymorphism (SNP) analyses, two major clusters were found for the colistin-resistant A. baumannii strains. However, no differences were observed by the time of detection for each cluster, the samples, the pattern of antibiotic resistance, or the patient characteristics. In the conventional MLST following the Oxford scheme, the typing result showed ST1809, ST451, ST191, ST1837, and ST369 in the global clone 2 (GC2), without any relation with the results of wgMLST and SNP analyses.ConclusionBased on the findings of the resistance gene analysis through WGS and comparative genomic analysis, the potential genes associated with colistin-resistance or CDS were examined. Furthermore, the analysis of molecular epidemiology through WGS regarding colistin-resistant A. baumannii may prove helpful in preventing infection by multidrug-resistant bacteria and controlling healthcare-associated infections.

Quantifying the effect of in-hospital antimicrobial use on the development of colistin-resistant Acinetobacter baumannii strains: a time series analysis

ObjectivesColistin is currently considered an essential therapeutic option for the treatment of hospital-acquired infections caused by resistant isolates of Acinetobacter baumannii. In this study, conducted in a tertiary care hospital,...

Diagnosis and Treatment of Bacterial Pneumonia in Critically Ill Patients with COVID-19 Using a Multiplex PCR Assay: A Large Italian Hospital's Five-Month Experience.

ABSTRACTBacterial pneumonia is a challenging coronavirus disease 2019 (COVID-19) complication for intensive care unit (ICU) clinicians. Upon its implementation, the FilmArray pneumonia plus (FA-PP) panel’s practicability for both the diagnosis and antimicrobial therapy management of bacterial pneumonia was assessed in ICU patients with COVID-19. Respiratory samples were collected from patients who were mechanically ventilated at the time bacterial etiology and antimicrobial resistance were determined using both standard-of-care (culture and antimicrobial susceptibility testing [AST]) and FA-PP panel testing methods. Changes to targeted and/or appropriate antimicrobial therapy were reviewed. We tested 212 samples from 150 patients suspected of bacterial pneumonia. Etiologically, 120 samples were positive by both methods, two samples were culture positive but FA-PP negative (i.e., negative for on-panel organisms), and 90 were negative by both methods. FA-PP detected no culture-growing organisms (mostly Staphylococcus aureus or Pseudomonas aeruginosa) in 19 of 120 samples or antimicrobial resistance genes in two culture-negative samples for S. aureus organisms. Fifty-nine (27.8%) of 212 samples were from empirically treated patients. Antibiotics were discontinued in 5 (33.3%) of 15 patients with FA-PP-negative samples and were escalated/deescalated in 39 (88.6%) of 44 patients with FA-PP-positive samples. Overall, antibiotics were initiated in 87 (72.5%) of 120 pneumonia episodes and were not administered in 80 (87.0%) of 92 nonpneumonia episodes. Antimicrobial-resistant organisms caused 78 (60.0%) of 120 episodes. Excluding 19 colistin-resistant Acinetobacter baumannii episodes, AST confirmed appropriate antibiotic receipt in 101 (84.2%) of 120 episodes for one or more FA-PP-detected organisms. Compared to standard-of-care testing, the FA-PP panel may be of great value in the management of COVID-19 patients at risk of developing bacterial pneumonia in the ICU.IMPORTANCE Since bacterial pneumonia is relatively frequent, suspicion of it in COVID-19 patients may prompt ICU clinicians to overuse (broad-spectrum) antibiotics, particularly when empirical antibiotics do not cover the suspected pathogen. We showed that a PCR-based, culture-independent laboratory assay allows not only accurate diagnosis but also streamlining of antimicrobial therapy for bacterial pneumonia episodes. We report on the actual implementation of rapid diagnostics and its real-life impact on patient treatment, which is a gain over previously published studies on the topic. A better understanding of the role of that or similar PCR assays in routine ICU practice may lead us to appreciate the effectiveness of their implementation during the COVID-19 pandemic.

Synergistic Activity of Capsaicin and Colistin Against Colistin-Resistant Acinetobacter baumannii: In Vitro/Vivo Efficacy and Mode of Action.

Acinetobacter baumannii is an opportunistic pathogen predominantly associated with nosocomial infections. With emerging resistance against polymyxins, synergistic combinations of drugs are being investigated as a new therapeutic approach. Capsaicin is a common constituent of the human diet and is widely used in traditional alternative medicines. The present study evaluated the antibacterial activities of capsaicin in combination with colistin against three unrelated colistin-resistant Acinetobacter baumannii strains in vitro and in vivo, and then further studied their synergistic mechanisms. Using the checkerboard technique and time-kill assays, capsaicin and colistin showed a synergistic effect on colistin-resistant A. baumannii. A mouse bacteremia model confirmed the in vivo effects of capsaicin and colistin. Mechanistic studies shown that capsaicin can inhibit the biofilm formation of both colistin-resistant and non-resistant A. baumannii. In addition, capsaicin decreased the production of intracellular ATP and disrupted the outer membrane of A. baumannii. In summary, the synergy between these drugs may enable a lower concentration of colistin to be used to treat A. baumannii infection, thereby reducing the dose-dependent side effects. Hence, capsaicin–colistin combination therapy may offer a new treatment option for the control of A. baumannii infection.

Prevalence and molecular mechanisms of colistin resistance in Acinetobacter baumannii clinical isolates in Tehran, Iran.

Colistin is one of the last remaining active antibiotics against multidrug resistant Gram-negative bacteria. However, several recent studies reported colistin-resistant (ColR) Acinetobacter baumannii from different countries. In the current study, we investigated molecular mechanisms involved in colistin resistance in A. baumannii isolates from different clinical samples.A total of 110 clinical A. baumannii isolates were collected from two hospitals in Tehran. Minimum inhibitory concentrations (MICs) were determined by broth microdilution according to the Clinical and Laboratory Standards Institute. For the ColR isolates, mutation was detected in pmrA, pmrB, lpxA, lpxC, and lpxD genes using the polymerase chain reaction (PCR) and sequencing. Moreover, the relative expression of the pmrC gene was calculated using quantitative reverse transcription PCR. Three colistin resistant isolates were identified with MIC between 8 and 16 μg/mL and were resistant to all the tested antimicrobial agents. All the three isolates had a mutation in the pmrB, pmrA, lpxA, lpxD, and lpxC genes. Moreover, the overexpression of pmrC gene was observed in all isolates. Our results showed that the upregulation of the PmrAB two component system was the primary mechanism linked to colistin resistance among the studied colistin resistant A. baumannii isolates.