High ESBL-E colonization rate among children in Gabon: a follow-up study.

A previous study conducted in Gabon, Central Africa, in 2010/11 found a high colonization rate with extended-spectrum ß-lactamase-producing enterobacterales (ESBL-E) among children of ~34 %. Eight years later, we aimed to reassess the ESBL-E rate and previously identified risk factors for colonization in children from Gabon. We conducted a cross-sectional cohort study in 2018 on 92 outpatients under 5 years of age with diarrhoea in Lambaréné, Gabon, in whom a rectal swab was obtained at the initial medical encounter (baseline). Fifty-eight of these provided a further rectal swab 1 week afterwards. ESBL-E colonization was assessed [following the European Committee on Antimicrobial Susceptibility Testing (EUCAST)], and in confirmed ESBL-E isolates the susceptibility to meropenem and the prevalence of the most abundant ESBL genes, bla CTX-M, bla SHV, and bla TEM, were investigated. At baseline, the ESBL-E colonization rate was 57 % (52/92; 95 % CI: 46-67). Hospitalization during the previous year, chicken consumption in the past week and young age were identified as independent risk factors for ESBL-E colonization at baseline. On day 7, the ESBL-E carriage rate was 72 % (42/58; 95 % CI: 59-83). All ESBL-E isolates (n=293) were susceptible to meropenem and bla CTX-M was the most frequently detected ß-lactamase gene. The ESBL-E colonization rate among children from Gabon is alarmingly high, with indications of further increase over recent years. While all ESBL-E strains remain currently susceptible to meropenem, in practice no adequate treatment is available locally for severe infections with such isolates. It is thus of the utmost importance to invest in improved hospital infection prevention and control measures to combat ESBL-E effectively.

Treatment for carbapenem-resistant Enterobacterales infections: recent advances and future directions.

Carbapenem-resistant Enterobacterales (CRE) are a growing threat to human health worldwide. CRE often carry multiple resistance genes that limit treatment options and require longer durations of therapy, are more costly to treat, and necessitate therapies with increased toxicities when compared with carbapenem-susceptible strains. Here, we provide an overview of the mechanisms of resistance in CRE, the epidemiology of CRE infections worldwide, and available treatment options for CRE. We review recentlyapproved agents for the treatment of CRE, including ceftazidime-avibactam, meropenem-vaborbactam, imipenem-relebactam, cefiderocol, and novel aminoglycosides and tetracyclines. We also discuss recent advances in phage therapy and antibiotics that are currently in development targeted to CRE. The potential for the development of resistance to these therapies remains high, and enhanced antimicrobial stewardship is imperative both to reduce the spread of CRE worldwide and to ensure continued access to efficacious treatment options.

Pressure response of carbapenems Klebsiella pneumoniae under antibiotic stress.

To analyze the drug-resistant phenotype and genetic characteristics of Carbapenem resistant Klebsiella pneumoniae (CRKP) in this region, and to study its different expression profiles in RNA level under the pressure of low levels of antibiotics. Trace dilution method and PCR method were used to detect the antibiotic resistance phenotype and antibiotic resistance gene carrying of CRKP strain, simulate the antibiotic stress process, and RNAseq was used to analyze the transcriptomic changes of CRKP strain. 37 CRKP strains, 27 Carbapenem sensitive Klebsiella pneumoniae (CSKP) CSKP strains and 42 sensitive strains were detected. The antibiotic resistance rate of CRKP strain was significantly higher than that of other drug-resistant strains, and there were many kinds of antibiotic resistance genes. Transcriptomic analysis showed that CRKP strain showed compensatory rise under meropenem stress at low concentration, and the expression of genes related to biofilm formation, pressure induction, pressure tolerance and transcriptional regulation was significantly changed. It was speculated that mrkAB, fimDH, phoHP and pspABCD clusters significantly altered their expression under the antibiotics stress response in CRKP strain. The detection rate of CRKP strain is high in this area. Under low levels of antibiotic stress, CRKP strain can not only survive by synthesizing antibiotic modified enzyme, but also respond by transcriptional regulation and biofilm changes, resulting in stress compensation. The discovery of this phenomenon explains the failure of treatment due to improper use of higher-order antibiotics from the perspective of genetic interaction.

Antimicrobial ability and mechanism analysis of Lactobacillus species against carbapenemase-producing Enterobacteriaceae.

BACKGROUND: This study aims to investigate the antimicrobial ability and mechanism analysis of Lactobacillus species against carbapenemase-producing Enterobacteriaceae (CPE). METHODS: Five Lactobacillus spp. strains and 18 CPE clinical isolates were collected. Their anti-CPE effects were assessed by agar well diffusion and broth microdilution assay, as well as time-kill test. Finally, the specific anti-CPE mechanism, especially for the effect of organic acids was determined using broth microdilution method. RESULTS: All of five Lactobacilli isolates displayed the potent activity against most CPE isolates with mean zones of inhibition ranging 10.2-21.1 mm. The anti-CPE activity was not affected by heating, catalase, and proteinase treatment. Under the concentration of 50% LUC0180 cell-free supernatant (CFS), lactic acid, and mix acid could totally inhibit the growth of carbapenem-resistant Klebsiella pneumoniae (CPE0011), and acetic acid could inhibit 67.8%. In contrast, succinic acid and citric acid could not inhibit the growth of CPE0011. While we decreased the concentration to 25%, only lactic acid and mix acid displayed 100% inhibition. In contrast, succinic acid, citric acid and acetic acid did not show any inhibitory effect. CONCLUSIONS: Lactobacillus strains exhibit potent anti-CPE activity, and lactic acid produced by Lactobacillus strains is the major antimicrobial mechanism.

Antimicrobial and Antibiofilm Activities of Citral Against Carbapenem-Resistant Enterobacter cloacae.

Citral that is produced in the essential oils of plants is an isomeric mixture of geranial and neral. Few recent studies exhibited robust antibacterial activity of citral against several pathogens. However, little is reported about effects of citral on carbapenem-resistant Enterobacter cloacae (CREC). The purpose of this study was to assess antibacterial and antibiofilm activities of citral against CREC. Here, minimum inhibitory concentrations (MICs) of citral against CREC were determined by the agar dilution method. Antibacterial mode of citral against CREC was elucidated by evaluating changes in intracellular adenosine triphosphate (ATP) concentration, intracellular pH (pHin), membrane potential, membrane integrity, and cell morphology. In addition, CREC cell damage within biofilms was examined by confocal laser scanning microscopy (CLSM). The results showed that the MIC of citral against CREC was 1000 µg/mL. Citral inhibited CREC growth and destroyed its cell membrane integrity, as measured by the decrease of intracellular ATP, pH, and membrane potential as well as distinctive deformation in cellular morphology. CLSM images demonstrated that citral could inactivate CREC cells within biofilms. These results revealed that citral exhibits potent antibacterial and antibiofilm activity against CREC, and thus has potential to be used as natural food preservatives to control CREC-associated infection spread.

Impact of BD Kiestra InoqulA streaking patterns on colony isolation and turnaround time of methicillin-resistant Staphylococcus aureus and carbapenem-resistant Enterobacterale surveillance samples.

OBJECTIVES: To determine if using alternative streaking patterns on the BD Kiestra InoqulA can impact colony isolation and improve turnaround time (TAT) of methicillin-resistant Staphylococcus aureus (MRSA) and carbapenem-resistant Enterobacterales (CRE) screening samples. METHODS: A total of 1571 positive MRSA screening samples were studied, of which 755 screening plates were streaked by the standard pattern (4-Quadrant uniform S200) and 816 plates were streaked by an alternative pattern (Zigzag 3.5-1 S200). A total of 424 CRE-positive screening samples were studied, of which 211 screening plates were streaked by the standard pattern (Zigzag 2.5-1 inoc S200) and 213 plates were streaked by an alternative customized pattern (Zigzag 3.5-1 vertstreak s200). RESULTS: There was a reduction in the number of MRSA screening plates with insufficient isolated colonies for confirmatory testing from 75 plates (9.9%) when using the standard pattern to 18 plates (2.2%) when using the alternative streaking pattern. MRSA cases with a TAT above 36 hours also reduced significantly from 144 (19.1%) to 20 (2.4%). The number of CRE screening plates with insufficient colonies for same-day confirmatory testing reduced from 16 (7.6%) when using the standard pattern to two plates (1.1%) when using the alternative customized pattern. CRE cases with a TAT above 36 hours also reduced from 16 (7.6%) to seven (3.3%). CONCLUSIONS: The change in streaking patterns resulted in more plates with sufficient isolated colonies as well as reduced man-hours and materials required to perform subculture of mixed colonies and overall improvements in TAT.

Changes in the prevalence of causative pathogens isolated from severe burn patients from 2012 to 2017.

Infection is the leading cause of mortality in severe burn patients, benefitting from periodic monitoring of changes in bacterial prevalence and antibiotic resistance trends. This single facility retrospective study evaluated blood culture results for patients hospitalized in the burn intensive care unit (BICU) from January 2012 to December 2017. A total of 969 samples from 420 patients were reviewed. Isolated pathogens were recorded by year and the number of days of hospitalization. Results showed that Acinetobacter baumanni was the most predominant isolated pathogen, followed closely by Pseudomonas aeruginosa, Klebsiella spp., and Enterococcus spp. Throughout this 6-year study, several significant trends were noted; Klebsiella species increased, while P. aeruginosa decreased. Staphylococcus aureus and Klebsiella species gradually increased, and P. aeruginosa doubled as the length of hospital stay increased to 22 days. Interestingly, as the length of the hospital stay increased, the proportion of Carbapenem-resistant Enterobacteriaceae (CRE) significantly increased up to 45.0% at 22 days (P=0.003). Conversely, the proportion of Acinetobacter baumannii gradually decreased as the days hospitalized increased. Overall, the rate of multidrug-resistant (MDR) bacteremia found in burn patients was substantially higher than that in other patients and appeared from the earliest phase of hospitalization. Therefore, early use of antibiotics targeting MDR Gram-negative bacteria in burn patients admitted to the BICU might be warranted. Further, since CRE infections increase in abundance over time, significant effort should be made to manage the initial CRE infections of burn patients before they can multiply into a life-threatening situation.

Biofilm-Forming by Carbapenem Resistant Enterobacteriaceae May Contribute to the Blood Stream Infection.

Bloodstream infection (BSI) due to carbapenem-resistant Enterobacteriaceae (CRE) has a high mortality rate and is a serious threat worldwide. Ten CRE strains (eight Enterobacter cloacae, one Klebsiella pneumoniae and one Citrobacter freundii) were isolated from the blood of nine patients, a percentage of whom had been treated with indwelling devices. The steps taken to establish cause included minimum inhibitory concentration (MIC) tests, a pulsed-field gel electrophoresis (PFGE), biofilm study, a multiplex PCR for resistant genes of carbapenemases and extended-spectrum beta-lactamases (ESBLs), and plasmid incompatibility typing. All strains showed a tendency toward resistance to multiple antibiotics, including carbapenems. Frequently isolated genes of ESBLs and carbapenemases include blaTEM-1 (four strains), blaSHV-12 (four strains) and blaIMP-1 (six strains). A molecular analysis by PFGE was used to divide the XbaI-digested genomic DNAs of 10 CRE strains into eight patterns, and the analysis showed that three E. cloacae strains detected from two patients were either identical or closely related. The biofilm production of all CRE strains was examined using a microtiter biofilm assay, and biofilm growth in continuous flow chambers was observed via the use of a confocal laser scanning microscope. Our study indicates that biofilm formation on indwelling devices may pose a risk of BSI due to CRE.

Weak biofilm formation among carbapenem-resistant Klebsiella pneumoniae.

Biofilm formation of multidrug and extensively drug resistant Klebsiella pneumoniae isolates is poorly understood. We investigated 139 diverse clinical K. pneumoniae isolates that possess various resistance patterns to evaluate the relationship between biofilm formation and resistance. Antimicrobial resistance was compared among a diverse collection of weak versus strong biofilm-forming K. pneumoniae, and predictors of strong biofilm formation were identified. Multi-drug resistant isolates were more common among weak (97.9%) versus strong biofilm formers (76%; P = 0.002). Carbapenem-resistant K. pneumoniae were 91% less likely to form strong biofilm (odds ratio 0.09; 95% confidence interval 0.02-0.33). The statistically significant inverse relationship between biofilm formation and antibiotic resistance suggests that virulence may be a trade-off for survival.

Efficacy of ceftazidime-avibactam in the treatment of infections due to Carbapenem-resistant Enterobacteriaceae.

BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) represent an important global threat. The aim of this study is to describe the clinical course and outcomes of patients with CRE infections treated with ceftazidime-avibactam (CAZ-AVI) compared to patients treated with other agents. METHODS: A retrospective cohort study of patients with established CRE infections from January 2017 until August 2018 was conducted. All patients who received CAZ-AVI and all cultures with carbapenem-resistant isolates were screened. We compared patients who received CAZ-AVI for CRE infections with patients who received other agents. RESULTS: A total of 38 consecutive patients with CRE infections were identified. Age and baseline comorbidities were similar between the two groups. The median time from admission to isolation of CRE culture was 22.5 days in the CAZ-AVI group and 17 days in the comparative group (P = 0.7). The incidence of CRE bacteremia was similar between the two groups: 7 patients (70%) in the CAZ-AVI group and 15 patients (53.6%) in the comparative group (P = 0.47). The most common type of CRE infections in both groups was hospital acquired pneumonia (HAP). Klebsiella pneumoniae was the predominant pathogen in both groups. A carbapenemase gene was detected in 35 (92%) patients; the OXA-48 gene was the predominant gene identified in 28 (74%) isolates. Eight out of ten patients in the CAZ-AVI group and fifteen out of twenty-eight in the comparative group achieved clinical remission (P = 0.14). After thirty days, all-cause mortality was observed in five patients in the CAZ-AVI group and 16 patients in the comparative group, accounting for 50 and 57% respectively. CONCLUSIONS: In patients with established OXA-48-type CRE infection, CAZ-AVI is a reasonable alternative to standard therapy. These findings need to be confirmed in prospective studies.

The potential of visible blue light (405 nm) as a novel decontamination strategy for carbapenemase-producing enterobacteriaceae (CPE).

Background: Carbapenemase-producing Enterobacteriaceae (CPE) pose a considerable threat to modern medicine. New treatment options and methods to limit spread need to be investigated. Blue light (BL) is intrinsically antimicrobial, and we have previously demonstrated significant antimicrobial effects on biofilms of a panel of isolates, including two CPEs.This study was performed to assess the antibacterial activity of 405 nm BL against a panel of CPE isolates (four encoding blaNDM, three blaKPC, two blaOXA-48, and three encoding both NDM and OXA-48 carbapenemases). Methods: In vitro experiments were conducted on 72 h old biofilms of CPEs which were exposed to 60 mW/cm2 of BL. Changes to biofilm seeding were assessed by measuring the optical density of treated and untreated biofilms. Results: Twelve bacterial clinical isolates (comprising eight Klebsiella pnemoniae, one K. oxytoca, and three Escherichia coli) were tested. BL was delivered for 5, 15 and 30 min, achieving doses of 162, 54, and 108 J/cm2, respectively.All of the CPEs were susceptible to BL treatment, with increasing reductions in seeding with increasing durations of exposure. At 30 min, reductions in biofilm seeding of ≥80% were observed for 11 of the 12 isolates, compared to five of 12 after 15 min. CPE_8180 was less susceptible than the rest, with a maximum reduction in seeding of 66% at 30 min. Conclusions: BL is effective at reducing the seeding of mature CPE biofilms in vitro, and offers great promise as a topical decontamination/treatment agent for both clinical and environmental applications.

Molecular and Phenotypic Study of Carbapenem Resistant E. coli.

BACKGROUND: Carbapenem resistance is an emerging problem. The aim of the present study was to determine the prevalence of carbapenem resistance and ESBLs among clinical isolates of E. coli by phenotypic methods and to study the molecular bases of the resistance by polymerase chain reaction (PCR). METHODS: The study was carried on 153 non repetitive E. coli strains collected from different clinical samples from Baghdad hospitals. The strains were isolated according to standard microbiological procedures and identified by Gram stain and biochemical reactions. E. coli strains were subjected to antimicrobial susceptibility tests by the disc diffusion method, carbapenem screening, ESBL screening tests, and molecular studies for genes responsible for such resistance. RESULTS: The study was carried out on 153 non repetitive E. coli. Detection of ESBLs by the double disc method reveals that 63 isolates (41.2%) of isolated E. coli had ESBL activity. Carbapenem resistance among E. coli reveals that 30 isolates (19.6%) had both metallo-ß-lactamase and carbapenemase activity as detected by double disc synergy test and boronic acid disc. Among 63 isolates of E. coli with positive double disc tests for ESBL resistance, the most frequent gene detected by PCR was blaTEM (52.4%), followed by blaSHV (33.3%) and blaCTX-M (14.3%). Among 30 E. coli strains with metallo-ß-lactamase activity and carbapenemase activity, the most frequent genes were IMP, VIM (30% for each), followed by NDM (20%), GIM, SIM, SPM, and KPC (10% for each). Isolated E. coli with carbapenem resistance represented 47.6% of E. coli with ESBL activity. CONCLUSIONS: The present study highlights the incidence of carbapenemase among clinical isolates of E. coli combined with extended ß-lactamases activity. Phenotypic screening methods were valuable for detection of different types of resistance. The common genes responsible for carbapenem resistance were IMP, VIM, and NDM. Further studies are recommended with more included E. coli isolates.

Meropenem-vaborbactam: a carbapenem and beta-lactamase inhibitor with activity against carbapenem-resistant Enterobacteriaceae.

Meropenem-vaborbactam is a carbapenem and ß-lactamase inhibitor combination that is newly indicated for the treatment of complicated urinary tract infections (cUTI), including adult pyelonephritis. Vaborbactam was developed due to emergence of carbapenem-resistant strains of Enterobacteriaceae. In a phase I trial, patients that received meropenem-vaborbactam 2-2 g intravenously over 3 h every 8 h, Cmax was 58.2 ± 10.8 µg/mL for meropenem and 59.0 ± 8.4 µg/mL for vaborbactam. AUC0-8 was 186 ± 33.6 µg â€¢ h/mL for meropenem and 204 ± 34.6 µg â€¢ h/mL for vaborbactam. Vss = 16.3 ± 2.6 L for meropenem and 17.6 ± 2.6 L for vaborbactam. Protein binding for vaborbactam averaged 33% in humans. Plasma clearance ranged from 10.42 ± 1.85 to 14.77 ± 2.84 L/h. One phase III trial evaluated efficacy for meropenem-vaborbactam 2-2 g intravenously every 8 h versus piperacillin-tazobactam 4-0.5 g intravenously every 8 h in complicated UTI. It found non-inferiority and statistical superiority for meropenem in overall success at the end of treatment primary end point. In another phase III trial evaluating efficacy in carbapenem-resistant Enterobacteriaceae (CRE) infections, meropenem-vaborbactam 2-2 g intravenously every 8 h was associated with decreased 28-day mortality and increased clinical cure compared with a best available therapy group.

Antibody-Mediated Killing of Carbapenem-Resistant ST258 Klebsiella pneumoniae by Human Neutrophils.

Carbapenem-resistant Klebsiella pneumoniae is a problem worldwide. A carbapenem-resistant K. pneumoniae lineage classified as multilocus sequence type 258 (ST258) is prominent in the health care setting in many regions of the world, including the United States. ST258 strains can be resistant to virtually all clinically useful antibiotics; treatment of infections caused by these organisms is difficult, and mortality is high. As a step toward promoting development of new therapeutics for ST258 infections, we tested the ability of rabbit antibodies specific for ST258 capsule polysaccharide to enhance human serum bactericidal activity and promote phagocytosis and killing of these bacteria by human neutrophils. We first demonstrated that an isogenic wzy deletion strain is significantly more susceptible to killing by human heparinized blood, serum, and neutrophils than a wild-type ST258 strain. Consistent with the importance of capsule as an immune evasion molecule, rabbit immune serum and purified IgG specific for ST258 capsule polysaccharide type 2 (CPS2) enhanced killing by human blood and serum in vitro Moreover, antibodies specific for CPS2 promoted phagocytosis and killing of ST258 by human neutrophils. Collectively, our findings suggest that ST258 CPS2 is a viable target for immunoprophylactics and/or therapeutics.IMPORTANCE Infections caused by carbapenem-resistant K. pneumoniae are difficult to treat, and mortality is high. New prophylactic approaches and/or therapeutic measures are needed to prevent or treat infections caused by these multidrug-resistant bacteria. A strain of carbapenem-resistant K. pneumoniae, classified by multilocus sequence typing as ST258, is present in many regions of the world and is the most prominent carbapenem-resistant K. pneumoniae lineage in the United States. Here we show that rabbit antibodies specific for capsule polysaccharide of ST258 significantly enhance human serum bactericidal activity and promote phagocytosis and killing of this pathogen by human neutrophils. These studies have provided strong support for the idea that development of an immunotherapy (vaccine) for carbapenem-resistant K. pneumoniae infections is feasible and has merit.

Impact of carbapenem-resistant Klebsiella pneumoniae (CR-KP) infections in kidney transplantation.

BACKGROUND: Carbapenem-resistant Klebsiella pneumoniae (CR-KP) infections in solid organ transplant patients are progressively increasing and are associated with worse outcomes, although potential risk factors and therapeutic strategies are still not well defined. METHODS: We conducted a retrospective matched-pair analysis in which we compared 26 recipients CR-KP-positive after kidney transplantation (KT) with 52 CR-KP-negative patients transplanted in the same period, during a CR-KP outbreak that occurred in our hospital. Twenty-one patients (80%) received a combined antibiotic treatment. At the end of the follow-up, of the 26 CR-KP infected patients, 11 (42.3%) experienced at least one episode of re-infection, 9 (34.6%) remained colonized, and 6 (23.0%) had a symptomatic infection. Two of the 11 patients with re-infection died, while 9 were colonized at the end of the study. RESULTS: A significantly better patient (P = .043) and graft (P < .001) survival was observed in CR-KP-negative patients. Univariate analysis identified the following variables as potential risk factors associated with CR-KP infection after KT: lower body mass index (P = .020); higher creatinine levels at post-transplant days 7 (P = .009), 15 (P = .026), and 30 (P = .019); longer hospital stay (P = .007); longer cold ischemia time (P = .004); delayed graft function (P = .020); and higher Clavien-Dindo score (P = .006). CONCLUSION: The study confirmed that a CR-KP positivity may affect the outcome of a kidney transplant population. In severe CR-KP infections with sepsis, a combined antibiotic treatment seems to be advisable.