Emergence Of Multidrug-resistant Acinetobacter Baumannii Research Articles

Role of Efflux Pump Gene adeIJK to Multidrug Resistance in Acinetobacter baumannii Clinical Isolates

Background: The emergence of multidrug-resistant Acinetobacter baumannii (MDRAB) as a nosocomial pathogen is one of the major public health problems. This study aimed to evaluate the role of an efflux pump gene adeJ for the multidrug resistance of A. baumannii clinical isolates. Methods: Two groups, MDRAB and susceptible A. baumannii (SAB), of baumannii clinical isolates were studied. The SAB group consisted of strains that did not meet the criteria of MDRAB and were susceptible to more categories of antibiotics than MDRAB. Antimicrobial susceptibility results obtained by the VITEKⅡ system were used in data analysis and bacterial group allocation. We performed real-time reverse transcription PCR to determine the relative expression of adeJ. We compared the relative expression of adeJ in comparison groups by considering two viewpoints: (1) MDRAB and SAB groups,and (2) susceptible and non-susceptible groups for each antibiotic used in this study.Results: The mean value of relative expression of adeJ of MDRAB and SAB groups was 1.4 and 0.92, respectively, and showed significant differences (P = 0.002). The mean values of relative expression of adeJ of susceptible and non-susceptible groups to the antibiotics cefepime, ceftazidime, ciprofloxacin, imipenem, meropenem, tigecycline, piperacillin/tazobactam, ticarcillin/clavulanic acid, trimethoprim/sulfamethoxazole, piperacillin, and gentamicin showed statistically significant differences (P < 0.05). Conclusion: The overexpression of adeIJK might contribute to the multi-drug resistance in A. baumannii clinical isolates. Further, the overexpression of adeIJK might be one of the factors contributing to the resistance to numerous antibiotics.

Emergence of multidrug-resistant Acinetobacter baumannii under fluctuating levofloxacin concentration and its control by chlorine and UV disinfection

Although wastewater treatment plants are considered hotspots for the spread of antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs), few studies have explored the mechanisms underlying the emergence of multidrug-resistance. Using four sequencing batch reactors under fluctuating concentrations of levofloxacin (LVX), we isolated 440 isolates, which were identified based on the sequence of the 16 S rRNA gene and growth at 37 ºC, focusing on potential human pathogens. We then evaluated the multidrug-resistance of 62% of these isolates by determining the minimum inhibition concentration (MIC) of erythromycin, tetracycline, chloramphenicol, and LVX in the emergence of ARB. The average MIC was increased more than 100-fold in the first exposure, whereas did not show the same response after re-exposure due to different LVX concentrations between the 2 conditions. We identified 33 Acinetobacter baumannii isolates that emerged only during re-exposure, especially in R-2 and R-3, in which the LVX concentration was changed from 4 and 16 mg/L, respectively, to 128 mg/L. Moreover, we assessed the disinfection efficiency of chlorine/UV treatment against pathogenic ARB. The growth of multidrug-resistant A. baumannii isolates was decreased by 0.9–7.4 log10 following disinfection treatment with > 0.5 mg /L Cl2 or > 30 min UV irradiation.

Multidrug-resistant Acinetobacter baumannii in healthcare settings in Africa

The emergence of multidrug-resistant Acinetobacter baumannii is a major concern to healthcare providers and facilities in many parts of the world. This bacterial pathogen is commonly implicated in hospital-acquired infections, particularly in critically ill patients admitted to the intensive care unit (ICU). The extensive use of antibiotics, particularly in ICUs, and the lack of proper infection control interventions in many hospitals have led to an increased emergence of multidrug-resistant A. baumannii. Infections due to multidrug-resistant A. baumannii are associated with prolonged hospital stays and high morbidity and mortality, particularly among hospitalized ICU patients. The lack of antibiotic stewardship programmes in many healthcare facilities has exacerbated the burden of A. baumannii infections in many parts of Africa. This review discusses the prevalence and antibiotic-resistance pattern of the multidrug-resistant A. baumannii, and the possible ways to address or minimise its emergence in healthcare settings in Africa.

Glycyrrhetinic acid protects against Multidrug-resistant Acinetobacter baumannii-induced lung epithelial cells injury by regulating inflammation and oxidative stress

Glycyrrhetinic acid (GA) is a bio-effective component of Licorice. The GA is a monomer and the ingredient is an Oleanane-type pentacyclic triterpenes that has been used as a remedy for years. Due to the abuse of antibiotics, people pay attention to the emergence of Multidrug-resistant Acinetobacter baumannii (MDR-AB). As a conditional pathogen, MDR-AB causes severe infection, endangering human lives. Our previous studies found GA played an important role in Yinhua Pinggan, a Chinese medicine. However, whether GA could protect lung epithelium from MDR-AB-induced cell injury was elusive. Herein, we investigated the effects of GA on MDR-AB-infected A549 cells. The results showed GA had slightly antibacterial activity to MDR-AB in the GA (high concentration) but no impact on drug resistance genes. Notwithstanding, GA could reverse MDR-AB-induced cell apoptosis, hampered adhesion and invasion of MDR-AB to cells, and inhibit pro-inflammatory cytokines expression of IL-1β, IL-6, and TNF. Besides, MDR-AB-induced reactive oxygen species, pro-oxidative protein malonaldehyde, and myeloperoxidase of cells were decreased by GA, while antioxidative proteins were recovered, showing antioxidative capacity of GA might play a critical role. The expressions of toll-like receptor (TLRs) - 1, 2, 4, 5, 6, and 9 were increased by MDR-AB infection, while GA reversed the tendency. Interestingly, GA inhibited MDR-AB induced myeloiddifferentiationfactor88 expression (MYD88), one downstream con-factors of TLRs, but no affection on Interferon regulatory Factor 3 (IRF3), the other one, indicating GA inhibited MDR-AB induced cell injury by impact TLR/MYD88 pathway to attenuate inflammation. Altogether, our results demonstrated that GA protects against MDR-AB-induced cell injury through its antioxidative and anti-inflammatory properties, which deserve further study in the future.

Transcriptome Analysis of Acinetobacter baumannii in Rapid Response to Subinhibitory Concentration of Minocycline.

The increasing emergence of multidrug-resistant Acinetobacter baumannii brings great threats to public health. Minocycline is a kind of semisynthetic derivative of the antibacterial drug tetracycline and is often used to treat infections caused by multidrug-resistant A. baumannii with other antibiotics. However, minocycline-resistant A. baumannii appears constantly. To rapidly explore the response of A. baumannii to minocycline stress, RNA-seq was carried out to compare the difference in the transcriptome of A. baumannii ATCC19606 in the presence or absence of minocycline. The results showed that 25 genes were differentially expressed, including 10 downregulated genes and 15 upregulated genes, and 24 sRNA were upregulated and 24 were downregulated based on the filter criteria (Log2FC > 1 or <−1 and FDR < 0.05). RtcB family protein and ABC transporter ATP-binding protein were upregulated by 2.6- and 11.3-fold, and molecular chaperone GroES, chaperonin GroL, class C beta-lactamase ADC-158, amino acid ABC transporter permease, and APC family permease were downregulated by at least two-fold in the presence of half-MIC minocycline. The differentially expressed genes are mainly involved in the stress response, the GroES/GroEL chaperonin system, and transport metabolic pathways. sRNA 1248 was significantly upregulated, and sRNA 1767, 5182, and 6984 were downregulated in a rapid response to minocycline. These results provide insights into the adaptive mechanism of A. baumannii to minocycline.

A Novel Acinetobacter baumannii Bacteriophage Endolysin LysAB54 With High Antibacterial Activity Against Multiple Gram-Negative Microbes.

The rapid spread and emergence of multidrug-resistant Acinetobacter baumannii and other pathogenic Gram-negative bacteria spurred scientists and clinicians to look for alternative therapeutic agents to conventional antibiotics. In the present study, an A. baumannii bacteriophage p54 was isolated and characterized. Morphological and genome analysis revealed that bacteriophage p54 belongs to Myoviridae family with a genome size of 165,813 bps. A novel endolysin, namely LysAB54, showing low similarity with other well-known related endolysins, was cloned, expressed, and characterized from the bacteriophage p54. LysAB54 showed significant bactericidal activity against multidrug-resistant A. baumannii and other Gram-negative bacteria, including Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli, in the absence of outer membrane permeabilizers. Based on all those observations, LysAB54 could represent a potential agent for the treatment of multidrug-resistant Gram-negative superbugs.

Efficacy of Mastoparan-AF alone and in combination with clinically used antibiotics on nosocomial multidrug-resistant Acinetobacter baumannii.

Emergence of multidrug-resistant Acinetobacter baumannii (MDRAB) has become a critical clinical problem worldwide and limited therapeutic options for infectious diseases caused by MDRAB. Therefore, there is an urgent need for the development of new antimicrobial agents or alternative therapy to combat MDRAB infection. The aim of this study was to investigate effects of Mastoparan-AF (MP-AF), an amphipathic peptide isolated from the hornet venom of Vespa affinis with broad-spectrum antimicrobial activity, on MDRAB. As compared with clinical used antibiotics, MP-AF exhibited potent antimicrobial activity at 2–16 μg/ml against the reference strain A. baumannii ATCC 15151 and seven MDRAB clinical isolates, especially the colistin-resistant MDRAB, E0158. The synergistic antimicrobial combination study revealed that MP-AF acted synergistically with specific antibiotics, e.g., ciprofloxacin, trimethoprim/sulfamethoxazole (SXT) or colistin against some isolates of the MDRAB. It was noteworthy when MP-AF combined with SXT exhibited synergistic activity against all SXT-resistant MDRAB isolates. The synergistic combination of MP-AF and antibiotics could reduce the dosage recommended of each antimicrobial agent and improve the safety of medications with ignorable adverse effects, such as colistin with nephrotoxicity in therapeutic dose. Furthermore, MP-AF combined with antibiotics with different antimicrobial mechanisms could reduce selective pressure of antibiotics on bacteria and prevent the emergence of antimicrobial-resistant strains. Importantly, we are the first finding that MP-AF could make MDRAB from the original non-susceptibility to SXT become sensitivity. In conclusion, MP-AF alone or in combination with other antibiotics, especially SXT, is a potential candidate against MDRAB infection in clinical medicine.

Correlation of Oxacillinase Gene Carriage With the Genetic Fingerprints of Imipenem-Resistant Clinical Isolates of Acinetobacter baumannii.

Background:Emergence of multidrug-resistant Acinetobacter baumannii has resulted in the treatment failure of related infections and an increase in patient mortality. The presence of class D β-lactamases (oxacillinases) in this organism is an important mechanism underlying resistance to beta-lactam antibiotics.Objectives:The aim of this work was to investigate the correlation between oxacillinase gene carriage and genetic fingerprints in imipenem-resistant burn and non-burn isolates of A. baumannii.Materials and Methods:Fifty-eight A. baumannii isolates were collected from October 2011 to April 2012, which included 28 burn isolates from Shahid Motahari Hospital and 30 non-burn isolates from Imam Hossein Hospital. The minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of imipenem were measured by the broth microdilution method. The presence of oxacillinase genes (OXA-23-, OXA-24-, OXA-51-, and OXA-58-like genes) was shown using type-specific primers and PCR. Genetic profiles were generated by RAPD-PCR fingerprinting.Results:OXA-23 was observed in 81% of the isolates and its distribution was similar within the two groups. The presence of OXA-51 was shown in 58.6% of the isolates, of which most were burn isolates (67.6%). OXA-24 was present in 20.7% of the isolates, all belonging to the burn group; OXA-58 was not observed in any of the isolates. RAPD-PCR fingerprints revealed two clusters at a similarity level of 70% (A, B). At a similarity level of 85%, nine different groups were observed for burn and non-bun isolates.Conclusions:Our results showed that blaOXA-23 was the most prevalent gene, followed by blaOXA-51, among the burn and non-burn clinical isolates of A. baumannii. BlaOXA-24-like genes were detected at a lower level and were only found among the burn isolates, which also showed higher heterogeneity compared to the non-burn group.

Antibiotic Resistance Profile and Distribution of Oxacillinase Genes Among Clinical Isolates of Acinetobacter baumannii in Shiraz Teaching Hospitals, 2012 - 2013

Background:The emergence of multidrug-resistant Acinetobacter baumannii complicates the therapy of the related infections. Hospital isolates of A. baumannii are usually multidrug-resistant. The problem is compounded by increasing resistance to broad-spectrum antibiotics including carbapenems.Objectives:The aim of this study was to determine antimicrobial susceptibility patterns and distribution of blaOXA-type carbapenemases genes among A. baumannii isolates from hospitalized patients in Shiraz, Southwest Iran.Materials and Methods:Two hundred A. baumannii isolates were recovered from different clinical specimens in four Shiraz teaching hospitals. Isolates were detected as A. baumannii by Microgen kit and PCR with specific primers of blaOXA-51-like gene. Antimicrobial susceptibility testing was determined by disk diffusion method for all the isolates. Multiplex PCR assays were performed for detection of blaOXA-23-like, blaOXA-24-like and blaOXA-58-like genes.Results:All the isolates were susceptible to colistin and polymyxin B. Moreover, all of them were resistant to piperacillin, piperacillin-tazobactam, ampicillin, ceftazidime, cefoxitin and aztreonam. Eighty (40%) isolates had positive results for blaOXA-23-like, 14 (7%) for blaOXA-24-like and 1 (0.5%) isolate for blaOXA-58-like. The co-existence of studied genes was detected for blaOXA-23-like plus blaOXA-24-like in nine (4.5%) isolates.Conclusions:The prevalence of carbapenem resistant A. baumannii isolates in Shiraz hospitals is high. The blaOXA-23-like gene was the most frequent carbapenemase identified among resistant A. baumannii isolated in Shiraz hospitals. The increasing incidence of A. baumannii is a serious concern, therefore control of this pathogen and taking preventive measures are emphasized.

Prevalence of eight resistance-nodulation-division efflux pump genes in epidemiologically characterized Acinetobacter baumannii of worldwide origin.

The increasing emergence of multidrug-resistant Acinetobacter baumannii constitutes a worldwide threat in hospital settings. Efflux-mediated resistance, particularly the resistance-nodulation-division (RND)-type efflux pumps, contributes significantly to decreased susceptibility to multiple antibiotics when overexpressed. Using PCR-based detection, the prevalence of genes encoding the RND efflux pumps AdeB, AdeJ and AdeG was investigated amongst 144 epidemiologically characterized and geographically diverse A. baumannii isolates of worldwide origin, representing International Clones 1-8 and genotypically unique isolates. Furthermore, five putative RND-type efflux genes identified via an in silico approach were included. Five of the eight investigated efflux pump genes were present in all isolates, including adeJ and adeG; the prevalence of the others varied between 65 and 97%. No association between the presence of one or multiple pumps to a specific clonal lineage was detected. The high prevalence of the efflux pump genes supports a fixed function of each individual pump that is not yet fully understood.

Tracing the emergence of multidrug-resistant Acinetobacter baumannii in a Taiwanese hospital by evaluating the presence of integron gene intI1.

BackgroundIn Changhua County, Taiwan, the number of clinical Acinetobacter baumannii isolates has risen since 2002, and multidrug-resistant Acinetobacter baumannii (MDRAB) has spread rapidly throughout Taiwan. In this study, to reveal the mechanism involved with the rapid dissemination of MDRAB emergence, the utility of the class 1 integron, intI1 integrase gene, as an MDRAB-associated biomarker was examined. A cross-sectional, clinical epidemiological study was performed at Changhua Christian Hospital between January 1st, 2001 and December 31st, 2004. Besides the existence of intI1 gene was examined, the pulse-field gel electrophoresis (PFGE) was also performed to determine the epidemiological characteristics of the isolates.FindingsThe overall hospital infection rate was 5–6%, while the infection rate of the intensive care unit (ICU) fluctuated. No positive correlation was observed between MDRAB isolates and the presence of intI1 (r = 0.168, P = 0.254). Additionally, no positive correlation was observed between the infection rate in the ICU and the presence of intI1 (r = -0.107, P = 0.468) or between the hospital infection rate and the presence of intI1 (r = -0.189, P = 0.199). However, two predominant clones among the MDRAB isolates were identified by PFGE.ConclusionsAlthough the presence of the intI1 gene does not seem suitable for tracing MDRAB emergence in Changhua County, two predominant clones were identified by PFGE, and subsequent studies to identify whether these clones were responsible for original nosocomial infection are needed.