Clinical Isolates Of Acinetobacter Baumannii Research Articles

Phenotypic Characterization and Heterogeneity among Modern Clinical Isolates of Acinetobacter baumannii.

Acinetobacter baumannii is an opportunistic pathogenic bacterium prioritized by WHO and CDC because of its increasing antibiotic resistance. Heterogeneity among strains represents the hallmark of A. baumannii bacteria. We wondered to what extent extensively used strains, so-called reference strains, reflect the dynamic nature and intrinsic heterogeneity of these bacteria. We analyzed multiple phenotypic traits of 43 nonredundant, modern, and multidrug-resistant, extensively drug-resistant, and pandrug-resistant clinical isolates and broadly used strains of A. baumannii. Comparison of these isolates at the genetic and phenotypic levels confirmed a high degree of heterogeneity. Importantly, we observed that a significant portion of modern clinical isolates strongly differs from several historically established strains in the light of colony morphology, cellular density, capsule production, natural transformability, and in vivo virulence. The significant differences between modern clinical isolates of A. baumannii and established strains could hamper the study of A. baumannii, especially concerning its virulence and resistance mechanisms. Hence, we propose a variable collection of modern clinical isolates that are characterized at the genetic and phenotypic levels, covering a wide range of the phenotypic spectrum, with six different macrocolony type groups, from avirulent to hypervirulent phenotypes, and with naturally noncapsulated to hypermucoid strains, with intermediate phenotypes as well. Strain-specific mechanistic observations remain interesting per se, and established "reference" strains have undoubtedly been shown to be very useful to study basic mechanisms of A. baumannii biology. However, any study based on a specific strain of A. baumannii should be compared to modern and clinically relevant isolates. IMPORTANCE Acinetobacter baumannii is a bacterium prioritized by the CDC and WHO because of its increasing antibiotic resistance, leading to treatment failures. The hallmark of this pathogen is the high heterogeneity observed among isolates, due to a very dynamic genome. In this context, we tested if a subset of broadly used isolates, considered "reference" strains, was reflecting the genetic and phenotypic diversity found among currently circulating clinical isolates. We observed that the so-called reference strains do not cover the whole diversity of the modern clinical isolates. While formerly established strains successfully generated a strong base of knowledge in the A. baumannii field and beyond, our study shows that a rational choice of strain, related to a specific biological question, should be taken into consideration. Any data obtained with historically established strains should also be compared to modern and clinically relevant isolates, especially concerning drug screening, resistance, and virulence contexts.

Detection of the blaVIM-2 Gene in Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates in Sudan

Background: Acinetobacter baumannii is a pleomorphic aerobic Gram-negative bacillus that is notorious for having multidrug resistance traits. Carbapenem-resistant Acinetobacter baumannii (CRAB) is a global concern due to its ability to retain and disseminate resistance genes, which poses a threat to the spread of resistance among bacterial communities in hospital settings. The aim of our study was to evaluate the existence of the blaVIM-2 gene in carbapenem-resistant Acinetobacter baumannii clinical isolates from Sudanese hospitals. Methods and Results: Forty clinical isolates of Acinetobacter baumannii were collected from June 2021 to April 2022. All isolates of Acinetobacter baumannii were identified via BioMérieux's Vitek-2 automated system (Marcy l'Étoile, France) and evaluated for phenotypic resistance to carbapenem, using imipenem and meropenem. The enzymatic mode of resistance was assessed by the Modified Hodge test (MHT). A real-time PCR was used to detect the presence of the blaVIM-2 gene. Of 40 isolates, 32(80%) were resistant to imipenem, 4(10%) were moderately resistant, and 4(10%) were susceptible to imipenem; 24(60%) were resistant to meropenem, 10(25%) were moderately resistant, and 6(15%) were susceptible to meropenem. MHT was 70% positive with imipenem use and 55% positive with meropenem use. Real-time PCR revealed that only 30% of the samples were positive for the blaVIM-2 gene. All blaVIM-2-positive isolates were resistant to both imipenem and meropenem. Conclusion: Resistance to carbapenem poses a serious threat, denying patients treatment options. It is essential to make continuous surveillance of these strains to prevent the development of resistant strains.

Synergetic dual antibiotics-loaded chitosan/poly (vinyl alcohol) nanofibers with sustained antibacterial delivery for treatment of XDR bacteria-infected wounds

Resistance of bacterial pathogens to conventional antibiotics has remained a significant challenge in managing post-wound infections, especially in developing countries. Here, a nanofibrous chitosan/poly (vinyl alcohol) (CS/PVA) mat was designed for controlled delivery of three different concentrations of two antibiotics (colistin/meropenem ratio of 32/64 μg/ml (AB1), 64/128 μg/ml (AB2), and 128/256 (AB3) μg/ml) with synergistic antibacterial activity against ATCC and extensively drug-resistant (XDR) Acinetobacter baumannii clinical isolates. The scaffolds showed a uniform fibrous structure with no bead formation with a sustained release of the antibiotics for one week. The elongation at break, wettability, porosity, and average fiber diameter decreased with increased antibiotics concentrations. Young's modulus and tensile strength showed a significant increase after adding antibiotics. All the constructs showed excellent in vitro cytocompatibility for fibroblasts and biocompatibility in an animal model. The antibacterial assays confirmed the dose-dependent antibacterial activity of the CS/PVA. The scaffolds loaded with AB2 and AB3 showed biocidal properties against ATCC, while only CS/PVA/AB3 had antibacterial activity against XDR clinical isolates. This study suggests the CS/PVA/AB3 nanofibrous scaffold contained 128/256 μg/ml colistin/meropenem as an excellent antibacterial wound dressing for protection of skin wounds from XDR clinical isolates and now promises to proceed with pre-clinical investigations.

SecReT6 update: a comprehensive resource of bacterial Type VI Secretion Systems.

Type VI Secretion System (T6SS) plays significant roles in microbial activities via injecting effectors into adjacent cells or environments. T6SS increasingly gained attention due to its important influence on pathogenesis, microbial competition, etc. T6SS-associated research is explosively expanding on numerous grounds that call for an efficient resource. The SecReT6 version 3 provides comprehensive information on T6SS and the interactions between T6SS and T6SS-related proteins such as T6SS regulators and T6SS effectors. To assist T6SS researches like microbial competition and regulatory mechanisms, SecReT6 v3 developed online tools for detection and analysis of T6SS and T6SS-related proteins and estimation of T6SS-dependent killing risk. We have identified a novel T6SS regulator and T6SS-dependent killing capacity in Acinetobacter baumannii clinical isolates with the aid of SecReT6 v3. 17,212 T6SSs and plentiful T6SS-related proteins in 26,573 bacterial complete genomes were also detected, analyzed and incorporated into the database. The database is freely available at https://bioinfo-mml.sjtu.edu.cn/SecReT6/ .

The evolution of carbapenem resistance determinants and major epidemiological lineages among carbapenem‐resistant Acinetobacter baumannii isolates in Germany, 2010‐2019

The aim of this study was to investigate and compare the molecular epidemiology and carbapenem resistance determinants in clinical Acinetobacter baumannii isolates collected during four multicentre surveillance studies conducted by the Paul-Ehrlich-Society for Infection Therapy. Isolates were collected prospectively from hospital in-patients at 17 medical centres in Germany over four periods of three- to six-months starting in October of each of 2010, 2013, 2016 and 2019. Species identification was performed by MALDI-TOF, gyrB multiplex polymerase chain reaction (PCR), and detection of the intrinsic blaOXA-51-like gene. Minimum inhibitory concentrations were determined by broth microdilution. The prevalence of carbapenemase-encoding genes was investigated by OXA-multiplex PCR and whole-genome sequencing. Molecular epidemiology was examined by rep-PCR and core-genome multi-locus sequence typing. A total of 302 A. baumannii isolates were collected. Resistance to imipenem and/or meropenem was detected in 58 isolates (19.2%) from 14 centres. The proportion of carbapenem-resistant isolates increased from 21.3% in 2010 to 33.3% in 2013, and then decreased to 13.8% in 2016 and 12.3% in 2019. Forty-six of these isolates were associated with the international clonal lineage IC2 and five with IC1. The most prevalent carbapenemase gene detected was blaOXA-23-like (n=51). Further carbapenem-resistance determinants were blaOXA-40-like (n=1), blaOXA-58-like (n=3) and blaNDM-1 (n=2). In one isolate, ISAba1 was detected upstream of blaOXA-51-like. In conclusion, IC2 was the most prevalent clonal lineage detected in this study. Interestingly, in Germany, carbapenem resistance seems to have decreased in A. baumannii between 2013 and 2019.

Molecular typing and antibiotic resistance patterns among clinical isolates of Acinetobacter baumannii recovered from burn patients in Tehran, Iran.

Acinetobacter baumannii (A. baumannii) is now considered a highly resistant pathogen to various types of antibiotics. Therefore, tracking the source of its prevalence and continuous control is crucial. This study aimed to determine antibiotic resistance and perform various molecular typing methods on clinical isolates of A. baumannii isolated from hospitalized burn patients in Shahid Motahari Burn Hospital, Tehran, Iran. Hospital isolates were confirmed by phenotypic and molecular methods. Then the sensitivity to different antibiotics was determined using the minimum inhibitory concentration (MIC) method. In order to perform molecular typing, three-locus dual assay multiplex polymerase chain reaction (PCR), multiple-locus variable-number tandem repeat analysis (MLVA), and multilocus sequence typing (MLST) methods were used. Among the 60 isolates collected, the frequencies of multidrug-resistant (MDR) and extensively drug-resistant (XDR) isolates were 90 and 10%, respectively. The most effective antibiotics were colistin with 100% and tigecycline with 83.33% sensitivity. Isolates were 100% resistant to piperacillin/tazobactam and cephalosporins, and 68.3% were resistant to carbapenem. The results of multiplex PCR showed five groups that international clone I (IC I) and IC II were the most common. The MLVA method identified 34 MLVA types (MTs), 5 clusters, and 25 singletons. Multilocus sequence typing results for tigecycline-resistant isolates showed seven different sequence types (STs). Increasing antibiotic resistance in A. baumannii isolates requires careful management to control and prevent the occurrence of the pre-antibiotic era. The results of this study confirm that the population structure of A. baumannii isolates has a high diversity. More extensive studies are needed in Iran to better understand the epidemiology of A. baumannii.

Evaluation of class 1, 2 and 3 integrons among multidrug resistant isolates of acinetobacter baumannii

The goal of this study was to determine the prevalence of class 1, 2, and 3 integrons in clinical Acinetobacter baumannii isolates from hospitalized patients. A total of 100 A. baumannii isolates were collected from several Imam Khomeini hospitals in Ilam, Iran. A. baumannii was discovered by traditional biochemical research. Following that, PCR was employed to detect several integron classes. Biochemical testing was used to identify Acinetobacter species. The samples obtained from patients (patients aged 21 to 69 years old, including 55 males and 45 women): Acinetobacter baumannii was found in 100 samples, Acinetobacter lufii in 25, and other Acinetobacter species in 12. 42 samples from the critical care unit were separated, 28 from the infectious department, 23 from the emergency department, and 7 from other departments. Furthermore, 44 blood samples, 23 trachea samples, 11 wound samples, 10 urine samples, and 12 unknown samples were isolated. Class 1 Integrons were found in 87 of 100 isolates, Class 2 Integrons in 74 of 100 isolates, and Class 3 Integrons in none of the isolates.

In vitro evaluation of the antimicrobial activity of antiseptics against clinical Acinetobacter baumannii strains isolated from combat wounds.

Healthcare-associated infections (HCAIs) are among the most prominent medical problems worldwide. In the context of increasing antibiotic resistance globally, the use of antiseptics as the main active agent and potentiator of antibiotics for the treatment of purulent-inflammatory complications of traumatic wounds, burns, and surgical wounds can be considered to tackle opportunistic infections and their prevention during war. This study presents a comparative investigation of the antimicrobial efficacy of antiseptics used for surgical antisepsis and antiseptic treatment of skin, mucous membranes, and wounds against multidrug-resistant clinical isolates of Acinetobacter baumannii as a wound pathogen of critical priority (according to the WHO). It was found that strains of A. baumannii, which have natural and acquired resistance to antimicrobial drugs, remain susceptible to modern antiseptics. Antiseptic drugs based on decamethoxine, chlorhexidine, octenidine, polyhexanide, and povidone-iodine 10% and 2% provide effective bactericidal activity against A. baumannii within the working concentrations of these drugs. Chlorhexidine and decamethoxine can inhibit biofilm formation by A. baumannii cells. In terms of bactericidal properties and biofilm formation inhibition, chlorhexidine and decamethoxine are the most effective of all tested antiseptics.

Antibiotic resistance and carriage class I integron in clinical isolates of Acinetobacter baumannii from Al Muthanna, Iraq.

The goal of this work was to systematically characterize and detect class 1, 2, and 3 integrons with many antibiotic resistance A. baumannii strains collected from a clinical environment in Iraq's Al-Muthanna hospitals. In this investigation, 24 non-replicated clinical strains of A. baumannii were evaluated using Chrome agar as a selective medium and PCR of the rplB gene. The clonal relatedness of the isolates to class 1 integron was evaluated using a PCR technique. The prevalence of class 1 integron was detected by PCR in only 12 clones of A. baumannii followed by HinfI digestion analysis showing three identical bands at 160 bp, 1350 bp, and 870 bp. In addition, PCR sequencing confirmed the presence of gene cassette arrays consisting of aacA4-catB8-aadA1 (100%) in class 1 integron. The sequence analysis of the integron shows 97.87 identity with A. baumannii isolates from Australia (GenBank accession number CP054302) among A. baumannii isolates. The blast analysis of this class I integron showed that the presence of the intI1, aacA4-catB8-aadA1 genes can considerably boost the acquisition of MDR phenotypes in A. baumannii isolates. We concluded that antibiotics of many types are widely used. The presence of integrons in A. baumannii is concerning for public health. In the clinical setting, it appears that the class 1 integron can be used as a predictive biomarker for the presence of MDR phenotypes. In these bacteria, however, the integron does not possess carbapenemases genes.

Role of Efflux Pumps in Reduced Susceptibility to Tigecycline among Clinical Isolates of Acinetobacter baumannii.

Acinetobacter baumannii (A. baumannii) is a very well-known emerging pathogen and has become a major burden on healthcare system especially in intensive care units (ICUs). Tigecycline is the last resort drug for treatment of multidrug-resistant A. baumannii infections. However, non-susceptibility to this drug is a rising problem. Resistance to tigecycline is mediated by Resistance-nodulation-cell division (RND) efflux pumps. This study was done to detect efflux pump genes (adeABC) and regulator genes (adeS,adeR) responsible for tigecycline resistance among the clinical isolates of A. baumannii. A total of 150 OXA-51 confirmed clinical isolates were subjected for tigecycline susceptibility test by broth microdilution (BMD) method. All isolates irrespective of their MIC were subjected to conventional PCR for detection of efflux genes (adeABC) and regulator genes (adeRS). Prevalence of tigecycline resistance was found to be 14 (9.33%) by the reference broth microdilution method (BMD). Overall prevalence of efflux genes was highest for adeB (69%) and lowest for adeR (29%). Combination of genes especially three, four or five were found more prevalent among resistant isolates with higher minimum inhibitory concentration (MIC). Combination of efflux genes confer higher MIC and can be a major contributor for resistance to tigecycline.

Engineering of lysin by fusion of antimicrobial peptide (cecropin A) enhances its antibacterial properties against multidrug-resistant Acinetobacter baumannii.

Most clinical isolates of Acinetobacter baumannii, a nosocomial pathogen, are multidrug-resistant (MDR), fueling the search for alternative therapies. Bacteriophage-derived endolysins have potent antibacterial activities and are considered as alternatives to antibiotics against A. baumannii infection. Gram-negative bacteria possess outer lipid membrane that prevents direct contact between the endolysins and the cell wall. We hypothesized that the fusion of antimicrobial peptide (AMP) with endolysin could help to reduce bacterial endolysin resistance and increase antimicrobial activity by membrane permeability action. Accordingly, we fused cecropin A, a commonly used AMP, with the N-terminus of AbEndolysin, which enhances the bactericidal activity of the chimeric endolysin. The bactericidal activity of cecropin A-fused AbEndolysin increased by at least 2–8 fold for various MDR A. baumannii clinical isolates. The in vitro bactericidal activity results also showed higher bacterial lysis by the chimeric endolysin than that by the parental lysin. The engineered AbEndolysin (eAbEndolysin) showed synergistic effects with the beta-lactam antibiotics cefotaxime, ceftazidime, and aztreonam, and an additive effect with meropenem and imipenem. eAbEndolysin had no cytotoxic effect on A549 cell line and rescued mice (40% survival rate) from systemic A. baumannii infection. Together, these findings suggest the potential of lysin therapy and may prompt its use as an alternative to antibiotics.

Genotypic and phenotypic comparison of clinical and environmental Acinetobacter baumannii strains

The genotypic and phenotypic characteristics and antibiotic resistance (antibiogram) profiles of clinical (n = 13) and environmental (n = 7) Acinetobacter baumannii isolates were compared. Based on the Repetitive Extragenic Palindromic Sequence-based PCR (REP-PCR) analysis, the clinical and environmental A. baumannii isolates shared low genetic relatedness (∼60%). Multilocus sequence typing (MLST, Oxford scheme) indicated that the clinical A. baumannii were assigned to three sequence types (ST231, ST945 and ST848), while the environmental A. baumannii (excluding AB 14) were categorised into the novel ST2520. The majority of the clinical (excluding AB 5, CAB 11, CAC 37) and environmental (excluding AB 14 and AB 16) A. baumannii strains were then capable of phase variation with both the translucent (71.4%; 15/21) and opaque (95.2%; 20/21) colony phenotypes detected. The clinical isolates however, exhibited significantly (p < 0.05) higher biofilm formation capabilities (OD570: 2.094 ± 0.497). Moreover, the clinical isolates exhibited significantly (p < 0.05) higher resistance to first line antibiotics, with 92.3% (12/13) characterised as extensively drug resistant (XDR), whereas environmental A. baumannii exhibited increased antibiotic susceptibility with only 57.1% (4/7) characterised as multidrug resistant (MDR). The environmental isolate AB 14 was however, characterised as XDR. In addition, only five clinical A. baumannii isolates exhibited colistin resistance (38.5%; 5/13). The current study highlighted the differences in the genotypic, phenotypic, and antibiotic resistance profiles of clinical and environmental A. baumannii. Moreover, the environmental strains were assigned to the novel ST2520, which substantiates the existence of this opportunistic pathogen in extra-hospital reservoirs.

Mechanisms of IS26-Mediated Amplification of the aphA1 Gene Leading to Tobramycin Resistance in an Acinetobacter baumannii Isolate.

ABSTRACTEnhanced levels of resistance to antibiotics arising from amplification of an antibiotic resistance gene that impact therapeutic options are increasingly observed. Amplification can also disclose novel phenotypes leading to treatment failure. However, the mechanism is poorly understood. Here, the route to amplification of the aphA1 kanamycin and neomycin resistance gene during tobramycin treatment of an Acinetobacter baumannii clinical isolate, leading to tobramycin resistance and treatment failure, was investigated. In the tobramycin-susceptible parent isolate, MRSN56, a single copy of aphA1 is present in the pseudocompound transposon PTn6020, bounded by directly oriented copies of IS26. For two clinical resistant isolates, new long-read sequence data were combined with available short-read data to complete the genomes. Comparison to the completed genome of MRSN56 revealed that, in both cases, IS26 had generated a circular translocatable unit (TU) containing PTn6020 and additional adjacent DNA. In one case, this TU was reincorporated into the second product generated by the deletion that formed the TU via the targeted conservative route and amplified about 7 times. In the second case, the TU was incorporated at a new location via the copy-in route and amplified about 65 times. Experimental amplification ex vivo by subjecting MRSN56 to tobramycin selection pressure yielded different TUs, which were incorporated at either the original location or a new location and amplified many times. The outcomes suggest that when IS26 is involved, amplification occurs via rolling circle replication of a newly formed TU coupled to the IS26-mediated TU formation or reincorporation step.IMPORTANCE Heteroresistance, a significant issue that is known to impact antibiotic treatment outcomes, is caused by the presence of spontaneously arising cells with elevated levels of resistance to therapeutically important antibiotics in a population of susceptible cells. Gene amplification is one well-documented cause of heteroresistance, but precisely how extensive amplification occurs is not understood. Here, we establish the case for the direct involvement of IS26 activity in the amplification of the aphA1 gene to disclose resistance to tobramycin. The aphA1 gene is usually found associated with IS26 in Gram-negative pathogens and is commonly found in extensively resistant Acinetobacter baumannii strains. IS26 and related IS cause adjacent deletions, forming a nonreplicating circular molecule known as a translocatable unit (TU), and amplification via a rolling circle mechanism appears to be coupled to either IS26-mediated TU formation or reincorporation. Related IS found in Gram-positive pathogens may play a similar role.

Analysis of Orthogonal Efflux and Permeation Properties of Compounds Leads to the Discovery of New Efflux Pump Inhibitors.

Optimization of compound permeation into Gram-negative bacteria is one of the most challenging tasks in the development of antibacterial agents. Two permeability barriers─the passive diffusion barrier of the outer membrane (OM) and active drug efflux─act synergistically to protect cells from the antibacterial action of compounds. In Escherichia coli (E. coli) and relatives, these two barriers sieve compounds based on different physicochemical properties that are defined by their interactions with OM porins and efflux pumps, respectively. In this study, we critically tested the hypothesis that the best substrates and inhibitors of efflux pumps are compounds that can effectively permeate the OM and are available at relatively high concentrations in the periplasm. For this purpose, we filtered a large subset of the ZINC15 database of commercially available compounds for compounds containing a primary amine, a chemical feature known to facilitate the uptake through E. coli general porins. The assembled library was screened by ensemble docking to AcrA, the periplasmic component of the AcrAB-TolC efflux pump, followed by experimental testing of the top predicted binders for antibacterial activities, efflux recognition, and inhibition. We found that the filtered primary amine library is a rich source of compounds with efflux-inhibiting activities and identified efflux pump inhibitors with novel chemical scaffolds effective against E. coli AcrAB-TolC and efflux pumps of multidrug-resistant clinical isolates of Acinetobacter baumannii. However, primary amines are not required for the recognition of compounds by efflux pumps and their efflux-inhibitory activities.

In Vitro and In Vivo Antimicrobial Activity of the Novel Peptide OMN6 against Multidrug-Resistant Acinetobacter baumannii.

The rapid worldwide spread of antimicrobial resistance highlights the significant need for the development of innovative treatments to fight multidrug-resistant bacteria. This study describes the potent antimicrobial activity of the novel peptide OMN6 against a wide array of drug-resistant Acinetobacter baumannii clinical isolates. OMN6 prevented the growth of all tested isolates, regardless of any pre-existing resistance mechanisms. Moreover, in vitro serial-passaging studies demonstrated that no resistance developed against OMN6. Importantly, OMN6 was highly efficacious in treating animal models of lung and blood infections caused by multidrug-resistant A. baumannii. Taken together, these results point to OMN6 as a novel antimicrobial agent with the potential to treat life-threatening infections caused by multidrug-resistant A. baumannii avoiding resistance.

Detection of biofilm production and antimicrobial susceptibility in clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa

Acinetobacter baumannii and Pseudomonas aeruginosa are commensal which commonly colonize humans. As a result of their ubiquitous nature, reservoirs in hospital environment and resistance to many antimicrobial agents they are responsible for hospital – acquired infections. Additionally treatment of these infections is difficult because of the ability for biofilm formation. Aim of the paper was to determine the association between biofilm formation on medical devices and antibiotic resistance profile, compared to respiratory samples in clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa. Material and methods: The study comprised 50 clinical samples (36 from medical devices and 14 as а control group from respiratory secretions). Acinetobacter baumannii and Pseudomonas aeruginosa were identified by routine microbiological methods. Modification of the microtiter plate assay described by Stepanovic et al. was used to investigate the formation of biofilm. The antimicrobial susceptibility testing was performed according to EUCAST guidelines. Results: Of the 50 analyzed strains, 16 (32%) were non-biofilm producers, and 34 (68%) were producing biofilms. Out of these, 29 (58%) were from medical devices, and 5 (10%) from the control group. Acinetobacter baumannii showed biofilm formation in 19 (67.9%), of which 17 (60.7%) from medical devices, and 2 (7.1%) from control group. Pseudomonas aeruginosa produced biofilm in 15 (68.1%), of which 12 (54.5%) from medical devices, and 3 (13.6%) from the control group. Multidrug resistance was detected in 40 (80%). All strains of Acinetobacter baumannii were multidrug resistant (MDR). For Pseudomonas aeruginosa, 11 (73.3%) biofilm forming isolates were MDR, and 1 (14.2%) non-biofilm forming isolate was MDR. Conclusion: Biofilm production was higher in strains from medical devices. Eighty percent of isolates were MDR. This is a serious challenge for treatment of these hospital-acquired infections.

Genomic Analysis of a Strain Collection Containing Multidrug-, Extensively Drug-, Pandrug-, and Carbapenem-Resistant Modern Clinical Isolates of Acinetobacter baumannii.

ABSTRACTIn this study, we characterize a new collection that comprises multidrug-resistant (MDR), extensively drug-resistant (XDR), pandrug-resistant (PDR), and carbapenem-resistant modern clinical isolates of Acinetobacter baumannii collected from hospitals through national microbiological surveillance in Belgium. Bacterial isolates (n = 43) were subjected to whole-genome sequencing (WGS), combining Illumina (MiSeq) and Nanopore (MinION) technologies, from which high-quality genomes (chromosome and plasmids) were de novo assembled. Antimicrobial susceptibility testing was performed along with genome analyses, which identified intrinsic and acquired resistance determinants along with their genetic environments and vehicles. Furthermore, the bacterial isolates were compared to the most prevalent A. baumannii sequence type 2 (ST2) (Pasteur scheme) genomes available from the BIGSdb database. Of the 43 strains, 40 carried determinants of resistance to carbapenems; blaOXA-23 (n = 29) was the most abundant acquired antimicrobial resistance gene, with 39 isolates encoding at least two different types of OXA enzymes. According to the Pasteur scheme, the majority of the isolates were globally disseminated clones of ST2 (n = 25), while less frequent sequence types included ST636 (n = 6), ST1 (n = 4), ST85 and ST78 (n = 2 each), and ST604, ST215, ST158, and ST10 (n = 1 each). Using the Oxford typing scheme, we identified 22 STs, including two novel types (ST2454 and ST2455). While the majority (26/29) of blaOXA-23 genes were chromosomally carried, all blaOXA-72 genes were plasmid borne. Our results show the presence of high-risk clones of A. baumannii within Belgian health care facilities with frequent occurrences of genes encoding carbapenemases, highlighting the crucial need for constant surveillance.

Prevalence of Aminoglycoside and Carbapenemase Resistance Genes and Biofilm Formation among Clinical Isolates of Acinetobacter baumannii in Iran

Prevalence of Aminoglycoside and Carbapenemase Resistance Genes and Biofilm Formation among Clinical Isolates of Acinetobacter baumannii in Iran

Global update on the in vitro activity of tigecycline and comparators against isolates of Acinetobacter baumannii and rates of resistant phenotypes (2016–2018)

ObjectivesThis study presents 2016–2018 in vitro antimicrobial activity data and rates of resistant phenotypes for clinical isolates of Acinetobacter baumannii from Africa/Middle East, Asia/South Pacific, Europe, Latin America, and North America. MethodsA total of 4320 A. baumannii isolates were collected across all regions between 2016 and 2018. The in vitro antimicrobial activities of amikacin, colistin, levofloxacin, meropenem, and tigecycline were determined using the broth microdilution methodology of the Clinical and Laboratory Standards Institute. MICs were interpreted using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints (version 11.0). Rates of subsets that were resistant to amikacin, colistin, levofloxacin, and meropenem, according to EUCAST breakpoints, are also presented. ResultsIn each region, tigecycline and colistin were active against isolates of A. baumannii (MIC90 values, 1 or 2 mg/L) and the lowest rate of resistance was to colistin (1.2%–7.3%). The rates of resistance to the panel of agents were generally lower among A. baumannii from North America (1.3%–42.7%), compared with the other regions. Fewer than 11% of meropenem-resistant A. baumannii were also resistant to colistin. The rates of amikacin-, levofloxacin- and meropenem-resistant A. baumannii were lowest in North America and mostly higher in Africa/Middle East and Latin America. ConclusionIn each geographical region, tigecycline and colistin maintained good in vitro antimicrobial activity against isolates of A. baumannii, including antimicrobial-resistant subsets. The higher rates of meropenem-resistant isolates, particularly in Africa/Middle East and Latin America, require continued monitoring because of the scarcity of effective treatment options.

Molecular characterization of Clinical carbapenem-resistant Acinetobacter baumannii isolates from two tertiary care hospitals in Indonesia

Background: Carbapenem-resistant Acinetobacter baumannii (CRAB) is an important pathogenic bacterium that can cause nosocomial infection in hospitalized patients with various manifestations. The purposes of this recent study were to determine the prevalence, antimicrobial susceptibility profiles, carbapenemase-producing phenotypic and genotypic of CRAB in two tertiary care hospitals in Indonesia. Methods: A. baumannii isolates collected from patient’s clinical cultures in two tertiary care hospitals in Malang and Manado were included. Identifications of meropenem-resistant A. baumannii isolates with the vitek2® system results, followed by a sensitivity test using 10 µg imipenem antibiotic disc according to CLSI guidelines to fulfill the criteria as CRAB isolates. We assessed carbapenemase-production using mCIM and eCIM, and determined the presence of blaKPC, blaNDM, blaOXA-23 carbapenemase resistance genes using simplex PCR. Results: 73 CRAB were collected from hospitalized patients, of which 30 CRAB from Manado and 43 CRAB from Malang. The largest number of samples came from sputum and indicates that XDR has also occurred in all CRAB isolates. Carbapenemase-production test using mCIM obtained positive results on 29 samples (96.7%) and 42 samples (97.7%) in Manado and Malang, respectively. The eCIM showed metallo-β-lactamase was dominant in two tertiary care hospitals. The prevalence of carbapenemase resistance genes was obtained blaOXA-23 and blaNDM ranged between 60% - 90.7% and 3.3% - 4.6%, respectively. blaKPC gene was not detected. Conclusions: We showed that CRAB isolates positive result of carbapenemase-production and carbapenemase resistance genes of blaOXA-23 seem to be dominant in two tertiary care hospitals in Malang and Manado, Indonesia. A national prevention and surveillance system should be prepared to reduce and limiting transmission of CRAB isolates.