Multidrug-resistant Gram-negative Isolates Research Articles

P-2113. Development of a Local Multidrug-Resistant Gram-Negative Antibiogram

Abstract Background As gram-negative (GN) resistance continues to rise, it is imperative to understand the best empiric treatment options on a local or regional level. Traditional antibiograms including all isolates may be diluted by sensitive organisms that do not have broad spectrum agents automated for susceptibility testing. The purpose of this review was to develop a health system antibiogram and assess susceptibility rates specific to multidrug-resistant (MDR) GN organisms, including newer broad-spectrum agents that are not routinely reported on traditional antibiograms. Multidrug-Resistant Gram-Negative Antibiogram 2017-2023 Methods Data was collected from acute care facilities within a large community hospital system. MDR GN isolates including extended spectrum β-lactamase-producing (ESBL) Enterobacterales, carbapenem-resistant (CR) Enterobacterales, difficult-to-treat (DTR) Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, and Burkholderia cepacia complex from January 2017 to December 2023 were included. Susceptibility testing was completed using VITEK 2 system (bioMérieux), E-test or Kirby-Bauer disk diffusion. Interpretation of susceptibility data was based on CLSI M100 guidance. Results The MDR GN antibiogram is shown in Table 1. The most common isolates encountered were ESBL Escherichia coli (n=8797) and ESBL Klebsiella pneumoniae (n=1779). DTR P. aeruginosa had the lowest susceptibility across all agents; in comparison to CR P. aeruginosa isolates, ceftazidime/avibactam susceptibility was similar however ceftolozane/tazobactam susceptibility was lower. Notably, ceftazidime/avibactam (100%) retained activity for CR K. pneumoniae while CR E. coli had a lower than expected isolate count and susceptibility. S. maltophilia had lower reported susceptibility to trimethoprim/sulfamethoxazole and levofloxacin (84% and 86% respectively) than minocycline (95%) which is performed upon request via manual E-test. Conclusion Most MDR GN pathogens within a large community hospital system had reported activity to newer antimicrobial agents. Overall, our antibiogram aligns with treatment recommendations advised by IDSA 2023 Antimicrobial Resistance guidance. Next steps include additional data review for annual susceptibility trends. Disclosures All Authors: No reported disclosures

Synergistic effect of (Azadirachta indica) Neem extract on multi drug-resistant gram-negative isolates from clinical samples at a tertiary care center

The indiscriminate usage of antibiotics in treatment and animal feed has led to the development of pan and multi drug resistance. The developing resistance in addition to the mortality had skewed the minds of researchers in search of newer antimicrobial agents. Among the native plants, one with potent antibacterial activity is Neem (Azadirachta indica). The leaves, bark, flowers, fruits, seed, and gum of the neem tree are utilised for various beneficial effects. The clinical samples including pus, urine, aspirate, sputum was sent to the microbiology department. The Gram negative Multi drug resistant organisms were included in the study. Around 210 isolates were obtained during the study period out of which 66 (31.4%) isolates were MDR. The synergistic effect of Neem extract was evaluated which showed a significant change in the susceptibility as intermediate range to susceptible range, resistant to intermediate and resistant to sensitive. The Neem extract has better antibacterial effect however, multicentric studies must be carried out to identify the minimum inhibitory concentration against various isolates.

Updating the Antimicrobial Resistance Pattern among Critical Priority Pathogens in the Intensive Care Unit in Northern Iran Post COVID-19 Pandemic.

During the COVID-19 pandemic, the widespread and indiscriminate prescription of antibiotics led to a significant increase in antibiotic resistance and the emergence of multi-drug-resistant (MDR) strains. This study aims to evaluate the prevalence of antibiotic resistance in MDR Gram-negative isolates in the intensive care unit (ICU) of northern hospitals in Iran following the COVID-19 pandemic. This is a cross-sectional study. The samples were collected from patients with healthcare-associated infections at ICU of hospitals in northern Iran. Antimicrobial resistance was assessed using standard broth macrodilution, and resistance genes were accurately identified using the multiplex polymerase chain reaction method. The present study revealed that the ICU had the highest frequency of MDR Acinetobacter baumannii infections (32.1%) and the lowest frequency of E. coli infections (12.6%). The frequency of resistance genes of A. baumannii is as follows: bla OXA-51 (100%), ampC (99.12%), apA6 (90.35%), and bla NDM (69.30%). Co-amoxiclav showed a 100% resistance rate, while Piperacillin-tazobactam had the lowest resistance rate at 38.2%. This study identified a high prevalence of MDR A. baumannii in ICU patients with healthcare-associated infections at northern hospitals in Iran, following COVID-19. Recommended treatments include Piperacillin-tazobactam or Meropenem.

Asymptomatic nasopharyngeal carriage of multidrug resistant bacteria among children at University of Gondar Hospital Northwest Ethiopia Revealing Hidden Health Risks

Gram-negative bacteria in the nasopharynx can eventually invade bacteria-limited sites and cause serious illnesses such as meningitis, otitis media, and pneumonia. However, data related to the carriage of these bacteria in children attending outpatient departments in the study area are limited. To assess nasopharyngeal carriage, antibiotic susceptibility patterns, and associated factors of gram-negative bacteria among children attending the outpatient department at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia. A hospital-based cross-sectional study was conducted from September 1, 2023, to December 30, 2023. A total of 385 children aged 3 to 14 years were enrolled using a systematic random sampling technique. Sociodemographic and clinical data were collected using a semistructured questionnaire. A total of 385 nasopharyngeal samples were collected using a sterile specimen collection nasopharyngeal swab, transported using Amie’s transport medium, and subsequently inoculated on chocolate agar, blood agar, modified thayer martin agar, and MacConkey agar plates. Bacterial species were identified by colony morphology, Gram staining, and biochemical tests such as oxidase tests, satellitism tests, and carbohydrate utilization tests. An antibiotic susceptibility test was performed using the Kirby–Bauer and modified Kirby–Bauer methods on Mueller–Hinton agar plates. The data were entered into Epi-Data version 4.6.0.6 and exported to SPSS version 25 for analysis. The adjusted odds ratio at a 95% confidence interval with a P value of < 0.05 in the binary logistic regression model was considered to indicate statistical significance. The overall nasopharyngeal carriage of gram-negative bacteria was 146 (37.9%) (95% CI: 33.2–42.9). Among these, nonfastidious gram-negative bacteria represented 45 (11.7%), followed by M. catarrhalis 41 (10.6%), N. meningitidis 34 (8.8%), and H. influenzae 26 (6.8%). The isolates exhibited high resistance to tetracycline (85; 75.9%), trimethoprim-sulfamethoxazole (105; 71.9%), ampicillin (76; 67.9%), and amoxicillin/clavulanic acid (60; 69.8%) but high susceptibility to meropenem (122; 83.6%), gentamicin (73; 84.9%), and minocycline (87; 72.5%). There were 99 total multidrug-resistant strains (67.8%, 95% CI: 59.7–75.0). Male sex (AOR = 1.785, 95% CI: 1.102–2.892, P = 0.019), smoking (AOR = 2.675, 95% CI: 1.149–6.230, P = 0.022), and large family size (≥ 5) (AOR = 1.857, 95% CI: 1.140–3.023, P = 0.013) were risk factors for nasopharyngeal colonization. Increased nasopharyngeal colonization of multidrug-resistant gram-negative isolates was observed in this study. Gentamicin, minocycline, and meropenem were the most effective antibiotics for the tested isolates. Bacterial colonization increased with increasing family size, smoking status, and male sex. Therefore, a definitive diagnosis in the outpatient pediatric department should be based on culture and susceptibility test results.

Genome sequences of 70 multidrug-resistant Gram-negative isolates in high-risk neonates in the Northeast of Mexico.

Infections by multidrug-resistant pathogens are steadily increasing worldwide. A considerable proportion of neonatal intensive care admissions have a bacterial infection with multidrug-resistant bacteria during their hospital stay. In this work, we report draft genome sequences of 70 selected isolates from high-risk neonates in the Northeast of Mexico.

Colorimetric Loop-Mediated Isothermal Amplification Assays Accurately Detect blaOXA-23-like and ISAba1 Genes from Acinetobacter baumannii in Pure Cultures and Spiked Human Sera.

Carbapenem resistance in Acinetobacter baumannii is a critical global health threat attributed to transferrable carbapenemase genes. Carbapenemase genotyping using polymerase chain reaction (PCR) presents a challenge in resource-limited settings because of its technical requirements. This study designed new loop-mediated isothermal amplification (LAMP) primers using multiple sequence alignment-based workflows, validated the primer performance against multiple target variants in silico, and developed novel LAMP assays (LAntRN-OXA23 and LAntRN-ISAba1) to detect the transferable blaOXA-23-like carbapenemase genes and ISAba1 elements in pure cultures and A. baumannii-spiked serum samples. The designed LAMP primers bind to the conserved regions of their highly polymorphic targets, with their in silico performance comparable with other published primers. The in vitro LAMP assays (using 30 PCR-profiled A. baumannii and 10 standard multidrug-resistant gram-negative isolates) have 100% concordance with the PCR-positive clinical samples, limits of detection as low as 1 pg/µL (200 copies/µL), and specificities of 57.89-100%. Both assays produced positive results when testing DNA samples (extracted using a commercial kit) from blaOXA-23-like and ISAba1-blaOXA-51-like PCR-positive A. baumannii-spiked normal human sera (five set-ups per target). In summary, the LAMP assays accurately detected the target genes and have applications in infection management, control, and point-of-care testing in resource-limited healthcare settings.

A comparative evaluation of colistin Minimum Inhibitory Concentration determination by reference broth microdilution with other commonly used phenotypic methods in Multidrug-Resistant Gram-negative bacilli

Background: over the past decade, the dependency on colistin as a last resort antibiotic has driven the global emergence of colistin resistance among many bacterial species. This study comparatively evaluated the colistin Minimum Inhibitory Concentration (MIC) by various phenotypic methods, including the reference method of Broth Microdilution (BMD), other approved methods of Colistin Broth Disk Elution (CBDE), and Colistin Agar Test (CAT) and widely available method of Epsilometer Test (E-test) among Multidrug Resistant (MDR) Gram-negative bacteria. Methods: ninety Gram-negative bacterial isolates that were resistant to three or more classes of drugs (MDR) were included in the study. All the MDR isolates were subjected to colistin susceptibility determination by BMD, CBDE, CAT, and E-test. Results: amongst 1118 samples, 90 (8.05%) samples yielded MDR Gram-negative bacilli. All the MDR Gram-negative isolates were colistin intermediate by all four methods of phenotypic colistin susceptibility. Three Acinetobacter baumannii and two Klebsiella pneumoniae isolates that had MIC of 2 μg/mL by BMD, displayed MIC of &lt;1 μg/mL by CBDE and CAT. Three isolates (2 Citrobacter koseri and 1 Enterobacter spp.) showed higher MIC by the E-strip method in comparison to BMD. Conclusions: our study holds significance, as there is a paucity of data comparing the four phenotypic methods for colistin MIC determination; BMD is the most reliable, gold standard method, but it is labor-intensive and requires technical expertise. In the present study, CBDE and CAT methods showed good concordance with BMD, and are easy to perform with limited logistics. Thus, they can be used as an alternative to BMD. We found that even though the Etest method was less accurate, it can still be used with caution to exclude the possibility of colistin resistance.

Meropenem-Vaborbactam Activity against U.S. Multidrug-Resistant Enterobacterales Strains, Including Carbapenem-Resistant Isolates.

Carbapenems are a common first-line therapy for serious Gram-negative infections, but carbapenem-resistant Enterobacterales (CRE) isolates have become an urgent health concern. Klebsiella pneumoniae serine carbapenemases (KPCs) now have been disseminated worldwide and are endemic in many hospitals globally. Isolates producing metallo-β-lactamases (MBLs) or class D OXA-48 carbapenemases are also increasingly common in Europe, although they are less common in the United States. Meropenem-vaborbactam is a combination of the carbapenem meropenem and vaborbactam, which is a β-lactamase inhibitor with activity against serine carbapenemases, including KPC-producing isolates. We examined the susceptibility of U.S. multidrug-resistant (MDR) isolates to meropenem-vaborbactam. A total of 1,697 MDR Enterobacterales isolates were collected in 31 U.S. medical centers in 2016 to 2020. Susceptibility testing was performed using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method. Whole-genome sequencing was performed for all CRE strains (MIC values of >2 mg/L for imipenem or meropenem). The rate of susceptibility of all MDR Enterobacterales strains to meropenem-vaborbactam was 99.1%, and 86.2% of the isolates were susceptible to meropenem. There were 222 CRE isolates (13.1%). KPC was the most common carbapenemase (81.1%). Thirteen CRE isolates produced NDM (n = 7), VIM (n = 3), and/or OXA-48-like (n = 4) carbapenemases; 29 CRE isolates (13.1%) had no detected carbapenemase. The rate of susceptibility of all CRE strains to meropenem-vaborbactam was 93.2%, and the rate of susceptibility of the KPC-producing isolates to meropenem-vaborbactam was 98.9%. The primary carbapenemase in the United States continues to be KPC, while MBL and OXA-48-like carbapenemases remain uncommon. Overall, the rate of susceptibility of these U.S. MDR organisms to meropenem-vaborbactam was 99.1%, indicating that meropenem-vaborbactam is a valuable treatment option for Gram-negative infections caused by U.S. MDR organisms. IMPORTANCE Carbapenems are a common first-line therapy for serious Gram-negative infections, but CRE isolates have become an urgent health concern. Meropenem-vaborbactam is a combination of the carbapenem meropenem and vaborbactam, which is a β-lactamase inhibitor with activity against serine carbapenemases, including KPC-producing isolates. We examined the susceptibility of U.S. MDR Gram-negative isolates to meropenem-vaborbactam. A total of 1,697 U.S. MDR Enterobacterales isolates collected in 2016 to 2020 were tested. Susceptibility testing was performed using the CLSI broth microdilution method. Whole-genome sequencing was performed for all CRE strains (MIC values of >2 mg/L for imipenem or meropenem). The rate of susceptibility of all MDR Enterobacterales strains to meropenem-vaborbactam was 99.1%, and 86.2% of the isolates were susceptible to meropenem. A total of 13.1% of the isolates were CRE strains, and KPC was the most common carbapenemase. Overall, the rate of susceptibility of these U.S. MDR organisms to meropenem-vaborbactam indicates that meropenem-vaborbactam is a valuable treatment option for Gram-negative infections caused by U.S. MDR Gram-negative pathogens.

Molecular surveillance of multidrug-resistant Gram-negative bacteria in Ukrainian patients, Germany, March to June 2022.

BackgroundSince the beginning of the war in Ukraine in February 2022, Ukrainians have been seeking shelter in other European countries.AimWe aimed to investigate the prevalence and the molecular epidemiology of multidrug-resistant Gram-negative (MDRGN) bacteria and meticillin-resistant Staphylococcus aureus (MRSA) in Ukrainian patients at admittance to the University Hospital Frankfurt, Germany.MethodsWe performed screening and observational analysis of all patients from March until June 2022. Genomes of MDRGN isolates were analysed for antimicrobial resistance, virulence genes and phylogenetic relatedness.ResultsWe included 103 patients (median age: 39 ± 23.7 years), 57 of whom were female (55.3%; 95% confidence interval (CI): 45.2-5.1). Patients were most frequently admitted to the Department of Paediatrics (29/103; 28.2%; 95% CI: 19.7-37.9). We found 34 MDRGN isolates in 17 of 103 patients (16.5%; 95% CI: 9.9-25.1). Ten patients carried 21 carbapenem-resistant (CR) bacteria, five of them more than one CR isolate. Four of six patients with war-related injuries carried eight CR isolates. In six of 10 patients, CR isolates caused infections. Genomic characterisation revealed that the CR isolates harboured at least one carbapenemase gene, bla NDM-1 being the most frequent (n = 10). Core genome and plasmid analysis revealed no epidemiological connection between most of these isolates. Hypervirulence marker genes were found in five of six Klebsiella pneumoniae CR isolates. No MRSA was found.ConclusionHospitals should consider infection control strategies to cover the elevated prevalence of MDRGN bacteria in Ukrainian patients with war-related injuries and/or hospital pre-treatment and to prevent the spread of hypervirulent CR isolates.

An assessment of minocycline sensitivity in multidrug-resistant gram-negative isolates in a tertiary care center of Western Maharashtra

An assessment of minocycline sensitivity in multidrug-resistant gram-negative isolates in a tertiary care center of Western Maharashtra

Mechanism of Action and Resistance Evasion of an Antimicrobial Oligomer against Multidrug-Resistant Gram-Negative Bacteria.

The last resort for treating multidrug-resistant (MDR) Pseudomonas aeruginosa and other MDR Gram-negative bacteria is a class of antibiotics called the polymyxins; however, polymyxin-resistant isolates have emerged. In response, antimicrobial peptides (AMPs) and their synthetic mimetics have been investigated as alternative therapeutic options. Oligothioetheramides (oligoTEAs) are a class of synthetic, sequence-defined oligomers composed of N-allylacrylamide monomers and an abiotic dithiol backbone that is resistant to serum degradation. Characteristic of other AMP mimetics, the precise balance between charge and hydrophobicity has afforded cationic oligoTEAs potent antimicrobial activity, particularly for the compound BDT-4G, which consists of a 1,4-butanedithiol backbone and guanidine pendant groups, the latter of which provides a cationic charge at physiological pH. However, the activity and mechanism of cationic oligoTEAs against MDR Gram-negative isolates have yet to be fully investigated. Herein, we demonstrated the potent antimicrobial activity of BDT-4G against clinical isolates of P. aeruginosa with a range of susceptibility profiles, assessed the kinetics of bactericidal activity, and further elucidated its mechanism of action. Activity was also evaluated against a panel of polymyxin-resistant isolates, including intrinsically-resistant species. We demonstrate that BDT-4G can evade some of the mechanisms conferring resistance to polymyxin B and thus may have therapeutic potential.

In Vitro Activity of Cefiderocol Against Multidrug-Resistant Gram-Negative Clinical Isolates

Background: New antimicrobials are being developed as a response to the global threat of multidrug-resistant and panresistant bacterial pathogens. Cefiderocol (FDC; Shionogi &amp; Co) is a novel parenteral siderophore cephalosporin with activity against gram-negative rods. Here, we report on the in vitro activity of FDC against multidrug-resistant gram-negative isolates collected by the CDC, including isolates available through the CDC and FDA Antibiotic Resistance Isolate Bank (AR Isolate Bank). Methods: The challenge set of gram-negative isolates (n = 339), most of which were obtained from the AR isolate bank (n = 258), comprised 188 Enterobacteriaceae (ENT), 72 Pseudomonas aeruginosa (PSA), and 79 Acinetobacter baumannii (ACB). Minimum inhibitory concentrations (MICs) for FDC in iron-depleted cation-adjusted Mueller-Hinton broth were determined using frozen reference broth microdilution panels (IHMA, Schaumburg, IL) according to CLSI guidelines. Isolates displaying nonsusceptibility to FDC (MIC &gt;4 µg/mL) underwent additional testing with β-lactamase inhibitors (FDC with 4 µg/mL avibactam, FDC with 100 µg/ml dipicolinic acid (DPA), and FDC with both 100 µg/mL dipicolinic acid (DPA) and 4 µg/mL avibactam). Results: Cefiderocol MICs ranged from ≤0.03 to &gt;64 µg/mL, and 313 (92.3%) isolates displayed susceptibility to FDC (MIC ≤4 μg/mL). The proportions of susceptible ENT, PSA, and ACB were 93.1%, 94.4%, and 88.6%, respectively. Among isolates harboring Ambler class A, class B, or class D carbapenemases, the proportions of susceptible isolates were 96.5%, 79.5%, and 94.0%, respectively. Overall, 26 (7.7%) isolates were categorized as FDC nonsusceptible (MIC ≥ 8 µg/mL); 65% of these were NDM producers. We selected 23 isolates for testing with β-lactamase inhibitors. The combination FDC-avibactam reduced the MIC to susceptible for all isolates harboring an Ambler class A or D carbapenemase, except for 1 OXA-71–producing ACB and 1 KPC-producing Citrobacter farmeri. The combination FDC-DPA reduced the MIC to susceptible for 9 of 13 (69.2%) NDM-producing and 4 of 4 (100%) OXA-23–producing ACB. By combining FDC with both DPA and avibactam, the MIC was reduced to susceptible (91%) for all but 1 KPC-producing and 1 NDM-producing Enterobacteriaceae isolate. Conclusions: Cefiderocol (FDC) demonstrated potent activity against a diverse collection of multidrug-resistant, gram-negative isolates, including producers of Ambler class A, B, and D carbapenemases. Among the 26 FDC nonsusceptible isolates, 65% were NDM positive. Our data indicate that FDC combined with β-lactamase inhibitors may restore susceptibility in FDC nonsusceptible isolates. Additional studies are needed to understand the underlying mechanism(s) of FDC resistance and to further explore the use of β-lactamase inhibitors in combination with FDC.Funding: NoneDisclosures: None

Cefiderocol as Adjunctive Treatment of Necrotizing Ventilator-associated Pneumonia Due to Extensively Drug-Resistant Pseudomonas aeruginosa

Abstract There are few treatment options for patients with extensively drug-resistant gram-negative infections. Cefiderocol is a novel β-lactam with excellent in vitro activity against multidrug-resistant gram-negative isolates, including carbapenem-resistant strains. However, this drug performed poorly versus best available therapy in an open-label randomized study of carbapenem-resistant gram-negative infections. Here, we present a case of a 32-year-old man with necrotizing ventilator-associated pneumonia caused by Pseudomonas aeruginosa who developed resistance to meropenem and ceftolozane/tazobactam while on therapy. Addition of expanded-access cefiderocol to his antimicrobial regimen contributed to successful microbiological cure while limiting exposure to nephrotoxic therapeutic agents.

Molecular characterization of multidrug-resistant Gram-negative pathogens in three tertiary hospitals in Cairo, Egypt

High rates of antimicrobial resistance (AMR) among Gram-negative pathogens (GNP) have been reported in Egypt. Antimicrobial surveillance and identifying the genetic basis of AMR provide important information to optimize patient care. In this study, we aimed to identify the beta-lactam resistance phenotypes and genotypes of multidrug-resistant (MDR) non-repetitive GNP from 3 tertiary hospitals in Egypt. WZe studied 495 non-repetitive MDR Gram-negative isolates from patients with complicated intra-abdominal infections (cIAI), complicated urinary tract infection (cUTI), and lower respiratory tract infection (LRTI), collected as part of the “Study for Monitoring Antimicrobial Resistance Trends” (SMART) conducted in 3 tertiary hospitals in Cairo, Egypt, from 2015 to 2016. Identification and susceptibility testing of GNP to antimicrobials were tested in each hospital laboratory and confirmed in a reference laboratory (International Health Management Associates (IHMA), Inc., Schaumburg, IL, USA). Molecular identification of extended-spectrum beta-lactamases (ESΒLs), AmpC, and carbapenem resistance genes was conducted in IHMA. Among the 495 MDR isolates, Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) were the most common (52.7% and 44.2%). K. pneumoniae was most susceptible to colistin, amikacin, ertapenem, and imipenem (92.7%, 72.7%, 69.3%, and 64%, respectively). E. coli was most susceptible to colistin (100%), amikacin (94.1%), imipenem (90.4%), and ertapenem (83.6%). ESBL was detected in 96.2% and ESBL genotypes included blaCTX-M-15 (70.1%), blaTEM-OSBL (48.5%), blaSHV-OSBL (27.9%), and blaCTX-M-14 (10.7%). AmpC resistance genes were identified in 9.7% of the isolates, dominated by blaCMY-2 (5.7%). Carbapenem resistance genes were detected in 45.3% of the isolates. In K. pneumoniae, blaOXA-48 dominated (40.6%), followed by blaNDM-1 (23.7%) and blaOXA-232 (4.5%). In E. coli, the most frequent genes were blaNDM-5 (9.6%), blaOXA-181 (5.5%), blaOXA-244 (3.7%), and blaNDM-1 (3.7%). blaKPC-2 was identified in 0.4% of isolates. Notably, 32.3% of isolates carried more than one resistance gene. Our findings emphasize the continued need for molecular surveillance of MDR pathogens, implementation of strict infection control measures, and antimicrobial stewardship policies in our hospitals.

In vitro evidence of the synergistic interaction of ceftopibrole and other antibiotics against multidrug-resistant Gram-negative isolates

In vitro evidence of the synergistic interaction of ceftopibrole and other antibiotics against multidrug-resistant Gram-negative isolates

Distribution, antimicrobial resistance and predictors of mortality in neonatal sepsis.

The aim of this study is to investigate etiological agents, patterns of antimicrobial resistance and predictors of mortality in culture proven neonatal sepsis. This is a twenty-four month retrospective cohort study of infants with culture proven sepsis. Demographic data, type of isolates and its sensitivity pattern were recorded. Multidrug resistant gram-negative isolates were defined as resistance to any three of five antibiotic classes: extended-spectrum cephalosporins, carbapenems, aminoglycosides, fluoroquinolones and piperacillin-tazobactam. A total of 183 case with culture positive sepsis were identified. Early onset sepsis occurred in 59% of cases. The majority of isolates (56.2%) were gram-positive but the most common individual isolates were klebsiella spp. (31.1%), Staphylococcus aureus (24.5%) and coagulase-negative staphylococci (CONS) (22.9%). The pathogen mix in early-onset did not differ from late-onset sepsis. High rates of multidrug resistance were observed in klebsiella spp. (49.1%), Escherichia coli (50%), citrobacter spp (50%), acinetobacter spp. (28.5%), pseudomonas spp. (100%) isolates. Methicillin resistance prevailed in 16.6% of coagulase-negative staphylococci, 24.4% of Staphylococcus aureus and 62.5% of enterococcus spp. Multivariate analysis revealed invasive ventilation and early onset sepsis to be independently associated with increased risk of mortality in contrast to breast milk feeding which is associated with decreased risk of mortality. A high degree of antimicrobial resistance underscores the need to understand the pathogenesis of resistance, curtail the irrational prescription of antibiotics in neonates and the requirement for measures to prevent it in low-income and middle-income countries.

In Vitro Activity of Cefiderocol against Multi-Drug Resistant Carbapenemase-Producing Gram-Negative Pathogens

BackgroundFew options remain for treatment of infections caused by multi-drug resistant (MDR), carbapenemase-producing gram-negative pathogens. Cefiderocol (CFDC; Shionogi & Co. Ltd), is a novel parenteral siderophore cephalosporin that enters the bacterial cell through the iron–siderophore uptake system. Here we report on the in vitro activity of CFDC against a set of well-characterized MDR gram-negative isolates collected by the Centers for Disease Control and Prevention.MethodsMinimum inhibitory concentrations (MIC) values for CFDC in iron-depleted cation-adjusted Mueller Hinton broth were determined using reference broth microdilution. Study isolates (n = 315) included Enterobacteriaceae (59%), Pseudomonas aeruginosa (19%), Acinetobacter baumannii (17%), Stenotrophomonas maltophilia (4%), and Burkholderia cepacia complex (1%). Of these, 229 (73%) were carbapenemase-producers including Ambler Class A- (37%), Class B- (29%) and Class D- type (29%) enzymes. The remaining isolates included 51 β-lactam-resistant isolates that were non-carbapenemase-producers, and 35 β-lactam-susceptible isolates. Results were interpreted using suggested CFDC breakpoints of Sensitive ≤4 μg/mL and Resistant ≥16 μg/mL.ResultsThe majority of the isolates (90.8%) were categorized as CFDC susceptible; the percentage of isolates with a CFDC MIC ≤4 μg/mL among Enterobacteriaceae, P. aeruginosa, and A. baumannii was 87.5%, 100%, and 89%, respectively. Percentage of isolates with a CFDC MIC ≤4 μg/mL that harbored a carbapenemase of the Class A-, Class B-, and Class D-type was 91.8%, 74.8%, 98.0%, respectively. By applying suggested breakpoints, 12 isolates were categorized as intermediate and 17 as resistant. The resistant isolates included 11 NDM-, 2 OXA-23- and 4 KPC-positive organisms.ConclusionCefiderocol showed potent activity against MDR gram-negative pathogens including Class A, B, and D carbapenemase-producing isolates. Of note, all P. aeruginosa, including Class B metallo-β-lactamase producers, were susceptible to CFDC.Disclosures All authors: No reported disclosures.

Activity of the novel siderophore cephalosporin cefiderocol against multidrug-resistant Gram-negative pathogens.

The novel siderophore cephalosporin cefiderocol (S-649266) with potent activity against Gram-negative pathogens was recently developed (Shionogi & Co., Ltd.). Here, we evaluated the activity of this new molecule and comparators against a collection of previously characterized Gram-negative isolates using broth microdilution panels. A total of 753 clinical multidrug-resistant Gram-negative isolates collected from hospitals worldwide were tested against cefiderocol and antibiotic comparators (ceftolozane-tazobactam [CT], meropenem [MEM], ceftazidime [CAZ], ceftazidime-avibactam [CZA], colistin [CST], aztreonam [ATM], amikacin [AMK], ciprofloxacin [CIP], cefepime [FEP], and tigecycline [TGC]) for their susceptibility. The collection included Escherichia coli (n=164), Klebsiella pneumoniae (n=298), Enterobacter sp. (n=159), Pseudomonas aeruginosa (n=45), and Acinetobacter baumannii (n=87). Resistance mechanisms included producers of carbapenemases and extended-spectrum β-lactamases (ESBLs). In addition, a series of colistin-resistant enterobacterial isolates (n=74), including 15 MCR-1 producers, were tested. The MIC90 of cefiderocol was 2mg/L, while those of comparative drugs were >64mg/L for CT, MEM, CAZ, CZA, and AMK, >32mg/L for ATM, >16mg/L for FEP, 8mg/L for CST, and 2mg/L for TGC. The MIC50 of cefiderocol was 0.5mg/L, while those of other drugs were >64mg/L for CAZ, 64mg/L for CT, >32mg/L for ATM, >16mg/L for FEP, 8mg/L for MEM and AMK, >4mg/L for CIP, 1mg/L for CZA, 0.5mg/L for TGC, and <0.5mg/L for CST. Only 20 out of 753 strains showed MIC values of cefiderocol ≥8μg/mL. Compared to the other drugs tested, cefiderocol was more active, with the exception of colistin and tigecycline showing equivalent activity against certain subgroups of bacteria.

Colonization of long term care facility patients with MDR-Gram-negatives during an Acinetobacter baumannii outbreak

BackgroundWe aimed to determine the prevalence of colonization by multidrug-resistant Gram-negative bacteria including ESBL-producing enterobacteriaceae, carbapenem-resistant enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii at two wards caring long term for patients with disorder of consciousness at the Geriatric Health Centers Graz, Austria. During our study we detected two A. baumannii outbreaks.MethodsIn August 2015, we conducted a point-prevalence study. Inguinal and perianal swabs were taken from 38 patients and screened for multidrug-resistant Gram-negative rods using standard procedures. Six months after the initial investigation all patients were sampled again and use of antibiotics during the past 6 months and mortality was registered. Genetic relatedness of bacteria was evaluated by DiversiLab system.ResultsFifty percent of patients were colonized by multidrug-resistant Gram-negative isolates. Five patients harboured ESBL-producing enterobacteriaceae. No carbapenem-resistant enterobacteriaceae were detected. 13/38 patients were colonized by A. baumannii isolates (resistant to ciprofloxacin but susceptible to carbapenems). There was a significant difference in the prevalence of colonization by A. baumannii between ward 2 and ward 1 (60% vs. 5.6%, p < 0.001). Two clusters of A. baumannii isolates were identified including one isolate detected on a chair in a patient’s room.ConclusionsWe detected a high prevalence of two multidrug-resistant A. baumannii strains in patients with disorder of consciousness at a LTCF in Graz, Austria. Our findings strongly suggest nosocomial cross-transmission between patients. An active surveillance strategy is warranted to avoid missing newly emerging pathogens.

Global Assessment of the Activity of Tigecycline against Multidrug-Resistant Gram-Negative Pathogens between 2004 and 2014 as Part of the Tigecycline Evaluation and Surveillance Trial.

Multidrug-resistant (MDR) Gram-negative organisms are a burden on the global health care system. The Tigecycline Evaluation and Surveillance Trial (TEST) is an ongoing global study designed to monitor the in vitro activities of tigecycline and a panel of marketed antimicrobials against a range of clinically significant pathogens. In this study, in vitro data are presented for MDR Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, and Enterobacter cloacae isolates collected from 2004 to 2014. In total, 13% (21,967/170,759) of isolates displayed multidrug resistance globally, with the highest rates recorded among A.baumannii (overall rate, 44% [8,294/18,741], increasing from 23% [309/1,323] in 2004 to 63% [447/712] in 2014). Other multidrug resistance rates ranged from 2.5% for K.oxytoca (203/8,000) to 12% for P.aeruginosa and K.pneumoniae (3,951/32,786 and 3,895/32,888, respectively), and rates among these pathogens remained stable during the study period. Against MDR E.coli, Klebsiella spp., and E.aerogenes, the lowest rates of resistance were to tigecycline (0.2%, 6%, and 12%, respectively), and the lowest MIC90 value against A.baumannii was observed for tigecycline (2mg/liter; MIC range, ≤0.008 to ≥32mg/liter). The only significant change in resistance to tigecycline during the study period was for MDR E.coli (P < 0.01), among which eight resistant isolates were identified globally from 2009 to 2013. In summary, these results show that tigecycline retained in vitro activity against the majority of MDR Gram-negative organisms presented here, but the rising rates of MDR A.baumannii highlight the need for the continued monitoring of global multidrug resistance. IMPORTANCE Multidrug resistance among bacterial pathogens is an ongoing global problem and renders antimicrobial agents ineffective at treating bacterial infections. In the health care setting, infections caused by multidrug-resistant (MDR) Gram-negative bacteria can cause increased mortality, longer hospital stays, and higher treatments costs. The aim of the Tigecycline Evaluation and Surveillance Trial (TEST) is to assess the in vitro antimicrobial activities of tigecycline and other contemporary agents against clinically relevant pathogens. This paper presents antimicrobial activity data from the TEST study between 2004 and 2014 and examines global rates of MDR Gram-negative isolates, including Acinetobacter baumannii, Pseudomonas aeruginosa, and members of the Enterobacteriaceae, during this time. Our results show that tigecycline retained in vitro activity against many MDR Gram-negative pathogens over the study period, while rates of MDR A.baumannii increased globally. Using these findings, we hope to highlight the current status of multidrug resistance in medical facilities worldwide.