Important Gram-negative Bacteria Research Articles

Enhanced sensing of bacteria biomarkers by ZnO and graphene oxide decorated PEDOT film

Developing a biofilm biomarker detector and inhibitor will immensely benefit efforts geared at curbing infectious diseases and microbiologically induced corrosion of medical implants, marine vessels and buried steel pipelines. N-Acyl homoserine lactones (AHLs) are important biomarkers gram-negative bacteria use for communication. In this work, we investigated the interactions between three AHL molecules and graphene oxide (GO) and ZnO nanomaterials embedded in conjugated poly(3,4-ethylenedioxythiophene) (PEDOT) film. The results show that PEDOT/GO/ZnO detected AHLs to a considerable extent with adsorption enthalpies of −4.02, −4.87 and −4.97 KJ/mol, respectively, for N-(2-oxotetrahydrofuran-3-yl)heptanamide (AHL1), 2-hydroxy-N-(2-oxotetrahydrofuran-3-yl)nonanamide (AHL2) and (E)-3-(3-hydroxyphenyl)-N-(2-oxotetrahydrofuran-3-yl)acrylamide (AHL3) molecules. The ZnO nanoparticles facilitated charge redistribution and transfer, thereby enhancing the conductivity and overall sensitivity of the substrate toward the AHLs. The adsorption distance and sites of interactions further tuned the charge migration and signal generation by the substrate, thus affirming the suitability of the modeled thin film as a sensor material. Excellent stability and conductivity were maintained before and after the adsorption of each AHL molecule. Moreover, the desorption time for each AHL molecule was calculated, and the result affirmed that the modeled film materials are promising for developing highly sensitive biosensors. Communicated by Ramaswamy H. Sarma

Essential Oil Quality of Lavender Grown Outside Its Native Distribution Range: A Study from Serbia

The aim of this study was to test the quality (physicochemical and sensory odor properties) and bioactivity (antimicrobial and antioxidant activities) of the essential oils (EO) obtained from the most frequently cultivated lavender and lavandin varieties in Serbia, whose cultivation areas were previously reserved for warmer climates, outside the agroecological region of Serbia. Seven EO from true lavenders (L. angustifolia Mill. and cultivars: ‘Hidcote blue’, ‘Munstead’, ‘Primorska’), Croatian indigenous lavandin cultivar (L. × intermedia ‘Budrovka’), lavandin ‘Grosso’ and one undetermined lavender sample (Lavandula sp.) showed compliance with standard requirements for lavender EO composition (contents of linalool 23.9–30.2% and 28.9–36.9%, and of linalyl acetate 22.2–32.2% and 6.9–20.7% in true lavender and lavandin samples, respectively). All EO were characterized as pleasant, with a floral aroma as a prominent odor. Samples exhibited high antimicrobial activities (3.5–14.2 µL mL−1 MIC and MBC values) against important Gram-positive (B. cereus and L. monocytogenes) and Gram-negative bacteria (E. coli) and yeasts (C. albicans), and high antioxidant capacity (IC50 values of 0.23–0.59 µg AAE mL−1 EO). This preliminary research on the quality of lavender EOs reveals the potential of this species for the future of medicinal and aromatic plant species production and further diversification of agriculture in the area.

In Vitro Anti-bacterial Effect of Ox-bile Against Some Important Gram-positive and Gram-negative Bacteria

Background: Ox-bile has been recommended as a natural remedy with several therapeutic potentials in traditional Persian medicine. It has had efficacy against inflammation and infection according to traditional medicine. Evidence revealed that bile disrupts bacterial cell membrane and degrades DNA structure, so it has anti-bacterial effects. However, there is no evidence of any approved medication composed of ox-bile in Iran. Objectives: The aim of this study was to evaluate the in vitro anti-bacterial effects of ox-bile. Methods: Ox-bile was obtained under aseptic conditions and sterilized with a 0.22 µm syringe filter, then examined for their sterility status through culture on different media. Following incubation under aerobic cultures for 48 hours and the anaerobic cultures for one week. Two different kinds of antimicrobial susceptibility tests, including well-diffusion method and serial dilution test were employed to characterize the inhibitory effect of ox-bile extraction on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes. Results: Based on our study, no anti-bacterial effect of ox-bile was observed against selected gram-positive and gram-negative bacteria. Conclusions: No in-vitro evidence of inhibitory effect was observed against studied gram-positive and gram-negative bacteria. Though further evaluation of the anti-bacterial effects of different preparations of ox-bile seems is still required.

A SERS bioassay based on vancomycin-modified PEI-interlayered nanocomposite and aptamer-functionalized SERS tags for synchronous detection of Acinetobacter baumannii and Klebsiella pneumoniae

Klebsiella pneumoniae (KP) and Acinetobacter baumannii (AB) are two important gram-negative bacteria that cause pneumonia and have been recently known to be associated with food. The rapid detection of these pathogens in food is important to minimize their colonization of the gut and stop new threats of the disease from spreading across the food chain. Herein, a double-edged sword aptasensor was developed for the synchronous detection of KP and AB in food and clinical samples. A highly sensitive, selective, specific, and synchronous detection of the target bacteria was achieved, and the limit of detection (LOD) was 10 cells/mL with a liner range of 50 to 105 cells/mL. The total assay time was 1.5 h. This study does not only provide a new tool for the detection of the target bacteria, but also serves as a promising tool for food safety and pneumonia diagnosis.

Carbapenem resistance in critically important human pathogens isolated from companion animals: a systematic literature review.

This study aimed to describe the presence and geographical distribution of Gram-negativebacteria considered critical on the priority list of antibiotic-resistant pathogens publishedby the World Health Organization, including carbapenem-resistant Enterobacteriaceae,carbapenem-resistant Acinetobacter spp., and carbapenem-resistant Pseudomonas aeruginosa.A systematic review of original studies published in 5 databases between 2010 and 2021 wasconducted, including genotypically confirmed carbapenem-resistant isolates obtained fromcanines, felines, and their settings. Fifty-one articles met the search criteria. Carbapenemresistant isolates were found in domestic canines and felines, pet food, and on veterinarymedical and household surfaces. The review found that the so-called "big five"-that is, the5 major carbapenemases identified worldwide in Enterobacterales (New Delhi metallo-βlactamase, active-on-imipenem, Verona integron-encoded metallo-β-lactamase, Klebsiellapneumoniae carbapenemase, and oxacillin [OXA]-48-like)-and the 3 most importantcarbapenemases from Acinetobacter spp. (OXA-23-like, OXA-40-like, and OXA-58-like) hadbeen detected in 8 species in the Enterobacteriaceae family and 5 species of glucose nonfermenting bacilli on 5 continents. Two publications used molecular analysis to confirmcarbapenem-resistant bacteria transmission between owners and dogs. Isolating criticallyimportant human carbapenem-resistant Gram-negative bacteria from domestic canines andfelines highlights the importance of including these animal species in surveillance programsand antimicrobial resistance containment plans as part of the One Health approach.

In vitro activity of cefiderocol against clinically important carbapenem non-susceptible Gram-negative bacteria from Saudi Arabia

Cefiderocol is a novel catechol-substituted siderophore cephalosporin with broad-spectrum activity against Gram-negative pathogens. However, variation of its activity among carbapenemase producers from various regions and countries has been reported. Here, we checked the in vitro activity against Gram-negative carbapenem non-susceptible bacteria collected in Saudi Arabia. Cefiderocol MICs were determined using the iron-depleted cation-adjusted Mueller-Hinton brothand interpreted according to the Clinical and Laboratory Standards Institute guidelines. Isolates (n=288) included carbapenemase-producing Escherichia coli (n=46), Klebsiella pneumoniae (n=98), Acinetobacter baumannii (n=65), and Pseudomonas aeruginosa (n=79) clinical isolates. Cefiderocol inhibited 73.26% (211/288) of the isolates studied at concentrations of ≤ 4 mg/L. Cefiderocol inhibited all carbapenem-resistant A. baumannii isolates (65/65, 100%) producing OXA-23-like, OXA-24-like, and NDM, and nearly all P. aeruginosa isolates (75/79, 94.94%), including those producing VIM and NDM. In contrast, the carbapenemase-producing isolates from the Enterobacterales group demonstrated significantly higher MICs with only 53.06% (52/98) of K. pneumoniae and 41.3% (19/46) of E. coli isolates exhibiting MICs of ≤4 mg/L. Isolates showing elevated MICs (73/144, 50.69%) included NDM (20/29, 68.97%), NDM/OXA-48-like (34/59, 57.63%), OXA-48-like (18/52, 34.62%), and KPC (1/4, 25%) producers, thus showing no clear association with the production of serine-type or metallo-type carbapenemases. However, high cefiderocol MICs (≥ 32mg/L) were associated with isolates producing NDM, and in particular, among those coproducing the OXA-232-type enzyme. Cefiderocol had excellent activity against multi-drug resistant non-fermenting Gram-negative pathogens. Reasons behind the high cefiderocol MICs in certain Enterobacterales isolates need further investigation.

Biological functions of sialic acid as a component of bacterial endotoxin.

Lipopolysaccharide (endotoxin, LPS) is an important Gram-negative bacteria antigen. LPS of some bacteria contains sialic acid (Neu5Ac) as a component of O-antigen (O-Ag), in this review we present an overview of bacteria in which the presence of Neu5Ac has been confirmed in their outer envelope and the possible ways that bacteria can acquire Neu5Ac. We explain the role of Neu5Ac in bacterial pathogenesis, and also involvement of Neu5Ac in bacterial evading the host innate immunity response and molecular mimicry phenomenon. We also highlight the role of sialic acid in the mechanism of bacterial resistance to action of serum complement. Despite a number of studies on involvement of Neu5Ac in bacterial pathogenesis many aspects of this phenomenon are still not understood.

Aeromonas allosaccharophila Strain AE59-TE2 Is Highly Antagonistic towards Multidrug-Resistant Human Pathogens, What Does Its Genome Tell Us?

Multidrug-resistant bacteria are of critical importance and a problem for human health and food preservation; the discovery of new antimicrobial substances to control their proliferation is part of the solution. This work reports on 57 antagonistic Aeromonas strains, of which 38 strains were antagonistic towards problematic human pathogens. The genome of the most antagonistic strain was sequenced and identified as Aeromonas allosaccharophila. Its genome was fully annotated and mined for genes that might explain that activity. Strain AE59-TE was antagonistic toward clinically relevant gram-negative and gram-positive multidrug-resistant bacteria, including Klebsiella pneumoniae KPC, Escherichia coli ESBL, Salmonella typhimurium, and Staphylococcus aureus MRSA. Strain AE59-TE2 was identified by multilocus sequence analysis. Genome mining identified four genes homologous to the bacteriocin, zoocin A from Streptococcus equi and a gene 98% similar to cvpA linked to colicin V production. A. allosaccharophila strain AE59-TE2 produced antimicrobial activity against a broad range of bacteria, including important gram-negative bacteria, not typically targeted by bacteriocins. Herewere described novel zoocin genes that are promising for industrial applications in the food and health sectors. Interesting and important antagonistic activity is described combined with the first detailed genomic analysis of the species Aeromonas allosaccharophila.

Thermosensitive hydrogel loaded with biosynthesized silver nanoparticles using Eucalyptus camaldulensis leaf extract as an alternative treatment for microbial biofilms and persistent cells in tissue infections

Biofilms play a key role in the development of chronic tissue infections and antimicrobial resistance. In situ-forming thermosensitive hydrogel containing biosynthesized silver nanoparticles using aqueous Eucalyptus camaldulensis leaf extract (bio-AgNPs) was developed as an efficient anti-biofilm agent with a sustained release for tissue infection treatment. The formulation with 24% poloxamer 407, 5% poloxamer 188, and 1% hydroxypropyl methylcellulose demonstrated gelation temperature at 33 °C and homogeneous dispersion of bio-AgNPs throughout three-dimensional networks. Pharmaceutical properties of bio-AgNPs-loaded hydrogel presented shear-thinning behavior and zero-order release profile. The formulation showed broad-spectrum antimicrobial activity and comparable effects with existing antibiotics against important pathogens, including Gram-positive, Gram-negative bacteria, and fungi. The formulation at 1/8–1/2 minimum inhibitory concentrations (MIC) significantly inhibited biofilm production in all tested pathogens (p < 0.05). Confocal laser scanning microscopy images clearly illustrated dead cells within mature biofilm upon the treatment with the formulation at 2MIC for 4 h. The formulation effectively scavenged free radicals and decreased nitric oxide production up to 98%. Decrease in mRNA levels of inflammation-related enzymes and pro-inflammatory cytokines in lipopolysaccharide-stimulated RAW 264.7 cells confirmed the formulation as a potential anti-inflammatory agent. The findings suggested that bio-AgNPs-loaded hydrogel represents a promising therapeutic approach for tissue infections.

Multicenter surveillance of antimicrobial susceptibilities and resistance mechanisms among Enterobacterales species and non-fermenting Gram-negative bacteria from different infection sources in Taiwan from 2016 to 2018

To explore the invitro antimicrobial susceptibility among clinically important Gram-negative bacteria (GNB) in Taiwan. From 2016 through 2018, a total of 5458 GNB isolates, including Escherichia coli (n=1545), Klebsiella pneumoniae (n=1255), Enterobacter species (n=259), Pseudomonas aeruginosa (n=1127), Acinetobacter baumannii complex (n=368), and Stenotrophomonas maltophilia (n=179), were collected. The susceptibility results were summarized by the breakpoints of minimum inhibitory concentration (MIC) of CLSI 2020, EUCAST 2020 (for colistin), or published articles (for ceftolozane/tazobactam). The resistance genes among multidrug-resistant (MDR) or extensively drug-resistant (XDR)-GNB were investigated by multiplex PCR. Significantly higher rates of non-susceptibility (NS) to ertapenem and carbapenemase production, predominantly KPC and OXA-48-like beta-lactamase, were observed in Enterobacterales isolates causing respiratory tract infection than those causing complicated urinary tract or intra-abdominal infection (12.7%/3.44% vs. 5.7%/0.76% or 7.7%/0.97%, respectively). Isolates of Enterobacter species showed higher rates of phenotypic extended-spectrum β-lactamase and NS to ertapenem than E. coli or K. pneumoniae isolates. Although moderate activity (54-83%) was observed against most potential AmpC-producing Enterobacterales isolates, ceftolozane/tazobactam exhibited poor invitro (44.7-47.4%) activity against phenotypic AmpC Enterobacter cloacae isolates. Additionally, 251 (22.3%) P. aeruginosa isolates exhibited the carbapenem-NS phenotype, and their MDR and XDR rate was 63.3% and 33.5%, respectively. Fifteen (75%) of twenty Burkholderia cenocepacia complex isolates were inhibited by ceftolozane/tazobactam at MICs of ≤4μg/mL. With the increase in antibiotic resistance in Taiwan, it is imperative to periodically monitor the susceptibility profiles of clinically important GNB.

Bacterial contamination on used face masks in healthcare personnel

Background: Face masks have been worn universally and for long periods of time by healthcare personnel during the COVID-19 pandemic. They are frequently touched or adjusted with the hands and may come in contact with various surfaces and high-touch sites when taken off and on even briefly. These activities present opportunities for face masks to become contaminated with microorganisms. Nursing homes have high rates of multidrug-resistant bacteria and low PPE compliance; therefore, contamination of face masks in this setting may be of great interest. We investigated bacterial colonization status on used face masks in healthcare personnel, including assessing the presence of clinically important and multidrug-resistant bacteria. Methods: At a nursing home serving mostly post–acute-care patients, we collected 69 face masks from personnel at the end of the user’s work shift. Information about the mask and the user was also collected via a self-reported survey. Face masks were incubated in BHI broth overnight at 36°C and 10 μL was then plated on selective and differential plates. Methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), and gram-negative bacteria (GNB) resistant to several antibiotic classes were identified using standard microbiological methods. Resistance testing for cefoxitin (S. aureus), ciprofloxacin, meropenem, tetracycline, erythromycin, gentamicin, trimethoprim–sulfamethoxazole, and ceftazidime with and without clavulanic acid (gram-negative bacteria) was performed using the disc diffusion technique on Mueller-Hinton plates (Kirby Bauer). Results: The job categories of face mask users were competency-evaluated nursing assistant or nursing assistant (22.73%), nurse (12.12%), and other or administrative (37.88%). Overall face mask contamination rates for MRSA (0%) and VRE (3.3%) were low; however, methicillin-susceptible S. aureus was found on 11 masks (15.9%). High contamination and resistance rates were found for gram-negative bacteria, with 113 isolates. Among them, 69 (60.9%) were resistant to at least 1 antibiotic, most commonly was erythromycin (59.4%). Additionally, higher rates of clinically important pathogenic gram-negative bacteria were identified: 14.3% of masks were contaminated with Klebsiella pneumoniae, 13.0% were contaminated with Enterobacter spp, and 4.2% were contaminated with Escherichia coli. Importantly, there were no significant differences in the total number of isolates of potential clinical significance recovered from masks worn &gt;6 hours versus those worn &lt;6 hours. Conclusions: Among nursing-home healthcare workers, face masks were often contaminated with multiple organisms, including potentially pathogenic bacteria and antibiotic-resistant gram-negative organisms. This contamination may pose a risk for transmission if face masks are not properly used and/or disposed of after wearing. Prolonged duration of face-mask wearing, however, was not associated with increased contamination rates.Funding: NoneDisclosures: None

Study of Antibacterial and Synergistic Activities of Cinnamomum verum, Eucalyptus camaldulensis and Zataria multiflora Boiss. in Persian Medicine Against Some Gram-Negative and Gram-positive Pathogenic Bacteria

Antibacterial activities of essential oils (EOs) of three widely used herbal elements of cinnamon (Cinnamomum verum J.Presl), eucalyptus (Eucalyptus camaldulensis Dehnh.), and thyme (Zataria multiflora Boiss.), whose antimicrobial effects are indirectly described in the old literature of Persian medicine as antiseptic agents and treatments of diseases caused by Havaye Vabaee (e.g., air-borne infectious diseases), were investigated. Antibacterial activities of the EOs were evaluated against some important Zoonosis and medicine pathogenic gram-negative (Escherichia coli, Klebsiella pneumonia, Listeria monocytogenes, and Pseudomonas aeruginosa) and gram-positive (Staphylococcus aureus, S. epidermidis, and Streptococcus pyogenes) bacteria. These pathogens cause disease in animals and humans.To this end, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and double-diffusion assay (DDA) tests were utilized using a microdilution method. While MIC values for cinnamon, eucalyptus, and thyme ranged between 0.25-1, 8-64, and 0.5-16 μg mL-1, respectively, the range of MBC values were 1-4, 32-256, and 2-64 μg mL-1. In DDA tests, a concentration of 0.5 μg mL-1 of the three EOs induced inhibition zones ranging between 20-35, 9-25, and 0-35 mm in diameter in the culture media of the above bacteria. Antibacterial effects of double combinations of the EOs against E. coli and S. aureus were also investigated by checkerboard test and fractional inhibitory concentration (FIC) index. Combinations of the EOs resulted in FIC index values of 1.06-2, indicating no interaction between most of the compounds combined. Our study also suggested the use of thyme oil with caution in combination with cinnamon or eucalyptus oils against E. coli because of their possible antagonistic effects (FIC index = 4).

Biosynthesis and Characterization of Silver Nanoparticles Produced by Phormidium ambiguum and Desertifilum tharense Cyanobacteria.

The world faces a challenge with the pervasion of multidrug-resistant bacteria that encourages scientists to develop and discover alternative, ecofriendly, and easy-to-produce new antibacterial agents. Our work is part of the greater effort of scientists around the world to achieve this goal by the biological synthesis of silver nanoparticles using cyanobacterial extracellular and intracellular components as nonchemical reducing agents. Two Egyptian cyanobacteria were isolated and identified according to 16S rRNA gene sequencing as Phormidium ambiguum and a novel species Desertifilum tharense. The sequences were deposited with accession numbers MW762709 and MW762710 for Desertifilum tharense and Phormidium ambiguum, respectively, in the GenBank. The results of UV-Vis analysis showed promising extracellular Ag-NPs synthesis by Desertifilum tharense and Phormidium ambiguum under light conditions. Therefore, these Ag-NPs were characterized and evaluated for antibacterial and antioxidant activity. TEM and SEM analyses revealed the spherical crystals with face-centered cubic structures and size range of 6.24–11.4 nm and 6.46–12.2 nm for Ag-NPs of Desertifilum tharense and Phormidium ambiguum, respectively. XRD and EDX results confirmed the successful synthesis of Ag-NPs in their oxide form or chloride form. The FTIR spectrum data confirmed the presence of hydroxyl and amide groups. Desertifilum tharense Ag-NPs displayed the largest inhibition zone that ranged from 9 mm against Micrococcus luteus ATCC 10240 to 25 mm against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300. For Phormidium ambiguum Ag-NPs, the inhibition zone diameter was in the range of 9 mm to 18 mm. The biosynthesized Ag-NPs significantly inhibited the growth of medically important resistance-pathogenic Gram-positive and Gram-negative bacteria. The Ag-NPs of Phormidium ambiguum exhibited the highest scavenging activity of 48.7% when compared with that of Desertifilum tharense, which displayed 43.753%.

Virulence factors of human pathogens: an always-needed approach / Fatores de virulência de patógenos humanos: uma abordagem sempre necessária

Hospital infections caused by resistant bacteria are a worldwide public health problem that mainly affects immunocompromised patients. These infections are mainly caused by inadequate antibiotic prescriptions, self-medication and long hospital stays. The World Health Organization mentions a list of resistant bacteria, especially Gram-negative ones considered as priorities for research, discovery and development of new drugs. These pathogens to survive and propagate use a varied set of strategies known as virulence factors. The present study corresponds to a review of the state-of-the-art related to main factors associated with pathogenicity in clinical important Gram-positive and Gram-negative bacteria. The virulence factors, alone or together, guarantee important defenses to the host organism's immune system, as well as adverse external conditions. Consequently, there is an increase in morbidity and mortality rates in hospital environments. Thus, effective microbial control measures are needed, especially in health institutions; also, the encouragement of new therapeutic approaches that target the main virulence factors.

Discovery of Small-Molecule VapC1 Nuclease Inhibitors by Virtual Screening and Scaffold Hopping from an Atomic Structure Revealing Protein-Protein Interactions with a Native VapB1 Inhibitor.

Nontypeable Haemophilus influenzae (NTHi) are clinically important Gram-negative bacteria that are responsible for various human mucosal diseases, including otitis media (OM). Recurrent OM caused by NTHi is common, and infections that recur less than 2 weeks following antimicrobial therapy are largely attributable to the recurrence of the same strain of bacteria. Toxin-antitoxin (TA) modules encoded by bacteria enable rapid responses to environmental stresses and are thought to facilitate growth arrest, persistence, and tolerance to antibiotics. The vapBC-1 locus of NTHi encodes a type II TA system, comprising the ribonuclease toxin VapC1 and its cognate antitoxin VapB1. The activity of VapC1 has been linked to the survival of NTHi during antibiotic treatment both in vivo and ex vivo. Therefore, inhibitors of VapC1 might serve as adjuvants to antibiotics, preventing NTHi from entering growth arrest and surviving; however, none have been reported to date. A truncated VapB1 peptide from a crystal structure of the VapBC-1 complex was used to generate pharmacophore queries to facilitate a scaffold hopping approach for the identification of small-molecule VapC1 inhibitors. The National Center for Advancing Translational Sciences small-molecule library was virtually screened using the shape-based method rapid overlay of chemical structures (ROCS), and the top-ranking hits were docked into the VapB1 binding pocket of VapC1. Two hundred virtual screening hits with the best docking scores were selected and tested in a biochemical VapC1 activity assay, which confirmed eight compounds as VapC1 inhibitors. An additional 60 compounds were selected with structural similarities to the confirmed VapC1 inhibitors, of which 20 inhibited VapC1 activity. Intracellular target engagement of five inhibitors was indicated by the destabilization of VapC1 within bacterial cells from a cellular thermal shift assay; however, no impact on bacterial growth was observed. Thus, this virtual screening and scaffold hopping approach enabled the discovery of VapC1 ribonuclease inhibitors that might serve as starting points for preclinical development.

Non-susceptibilities to antibiotics against important Gram-negative bacteria, and imipenem-relebactam, meropenem-vaborbactam against carbapenem non-susceptible Enterobacterales and Pseudomonas aeruginosa isolates implicated in complicated intra-abdominal and urinary tract infections in Taiwan, 2019

Susceptibility of isolates of top-ranking Enterobacterales species and Pseudomonas aeruginosa implicated in complicated intra-abdominal infections (cIAI) and urinary tract infections (cUTI) to important antibiotics, including imipenem-relebactam (IMR) and meropenem-vaborbactam (MVB), in Taiwan in 2019 were evaluated. MICs to various antibiotics were determined using broth microdilution method. Susceptibility results were interpreted mainly based on the MIC breakpoints of the Clinical and Laboratory Standards Institute (CLSI) 2021, but susceptibilities of IMR and MVB were interpreted based on the CLSI and European Committee on Antimicrobial Susceptibility Testing (EUCAST) 2021. Resistance genes amongst carbapenem-non-susceptible (NS) Gram-negative bacteria (GNB) were investigated using multiplex polymerase chain reaction (PCR). Escherichia coli (n=356), Klebsiella pneumoniae (n=165) and Enterobacter cloacae complex (n=42) isolates accounted for 85.3% of the 660 Enterobacterales isolates. The non-susceptibility rates of imipenem (IPM), IMR against isolates of non-Morganellaceae Enterobacterales, and meropenem (MEM), MVB against all Enterobacterales isolates were 92.2%/94.8%, 98.4-98.7%/98.4-99%, 95%/98.2% and 98.8-100%/99.4-100% for the cIAI/cUTI subgroups, respectively. Amongst the 40 IPM-NS-non-Morganellaceae Enterobacterales isolates, when the CLSI 2021 criteria were applied, 10 were NS to IMR, and four Klebsiella pneumoniae isolates (harbouring blaKPC but neither blaMBL nor blaOXA-48-like genes) were NS to IMR and MVB. Amongst the 93 Pseudomonas aeruginosa isolates under evaluation, the addition of relebactam (4mg/L) resulted in a 4-to-16-fold reduction in the MICs of IPM in all 15 IPM-NS-Pseudomonas aeruginosa isolates (including 10 porin-deficient ones) not harbouring blaMBL/blaOXA-48-like genes. Contrastingly, the addition of vaborbactam (8mg/L) improved the non-susceptibility to MEM in one (20%) of five IPM/MEM-NS Pseudomonas aeruginosa isolates. Continuous monitoring of susceptibility to clinically important GNB is warranted.

How fluoroquinolone preauthorization affects third- and fourth-generation cephalosporin use and resistance in a large academic hospital.

We observed an overall increase in the use of third- and fourth-generation cephalosporins after fluoroquinolone preauthorization was implemented. We examined the change in specific third- and fourth-generation cephalosporin use, and we sought to determine whether there was a consequent change in non-susceptibility of select Gram-negative bacterial isolates to these antibiotics. Retrospective quasi-experimental study. Academic hospital. Fluoroquinolone preauthorization was implemented in the hospital in October 2005. We used interrupted time series (ITS) Poisson regression models to examine trends in monthly rates of ceftriaxone, ceftazidime, and cefepime use and trends in yearly rates of nonsusceptible isolates (NSIs) of select Gram-negative bacteria before (1998-2004) and after (2006-2016) fluoroquinolone preauthorization was implemented. Rates of use of ceftriaxone and cefepime increased after fluoroquinolone preauthorization was implemented (ceftriaxone RR, 1.002; 95% CI, 1.002-1.003; P < .0001; cefepime RR, 1.003; 95% CI, 1.001-1.004; P = .0006), but ceftazidime use continued to decline (RR, 0.991, 95% CI, 0.990-0.992; P < .0001). Rates of ceftazidime and cefepime NSIs of Pseudomonas aeruginosa (ceftazidime RR, 0.937; 95% CI, 0.910-0.965, P < .0001; cefepime RR, 0.937; 95% CI, 0.912-0.963; P < .0001) declined after fluoroquinolone preauthorization was implemented. Rates of ceftazidime and cefepime NSIs of Enterobacter cloacae (ceftazidime RR, 1.116; 95% CI, 1.078-1.154; P < .0001; cefepime RR, 1.198; 95% CI, 1.112-1.291; P < .0001) and cefepime NSI of Acinetobacter baumannii (RR, 1.169; 95% CI, 1.081-1.263; P < .0001) were increasing before fluoroquinolone preauthorization was implemented but became stable thereafter: E. cloacae (ceftazidime RR, 0.987; 95% CI, 0.948-1.028; P = .531; cefepime RR, 0.990; 95% CI, 0.962-1.018; P = .461) and A. baumannii (cefepime RR, 0.972; 95% CI, 0.939-1.006; P = .100). Fluoroquinolone preauthorization may increase use of unrestricted third- and fourth-generation cephalosporins; however, we did not observe increased antimicrobial resistance to these agents, especially among clinically important Gram-negative bacteria known for hospital-acquired infections.

Pyelonephritis in Pediatric Uropathic Patients: Differences from Community-Acquired Ones and Therapeutic Protocol Considerations. A 10-Year Single-Center Retrospective Study.

Pyelonephritis (PN) represents an important cause of morbidity in the pediatric population, especially in uropathic patients. The aim of the study is to demonstrate differences between PNs of uropathic patients and PNs acquired in community in terms of uropathogens involved and antibiotic sensitivity; moreover, to identify a proper empiric therapeutic strategy. A retrospective study was conducted on antibiograms on urine cultures from PNs in vesicoureteral reflux (VUR) patients admitted to pediatric surgery department and from PNs in not VUR patients admitted to Pediatric Emergency Unit between 2010 and 2020. We recorded 58 PNs in 33 patients affected by VUR and 112 PNs in the not VUR group. The mean age of not VUR patients at the PN episode was 1.3 ± 2.6 years (range: 20 days of life–3 years), and almost all the urine cultures, 111 (99.1%), isolated Gram-negative bacteria and rarely, 1 (0.9%), Gram-positive bacteria. The Gram-negative uropathogens isolated were Escherichia coli (97%), Proteus mirabilis (2%), and Klebsiella spp. (1%). The only Gram-positive bacteria isolated was an Enterococcus faecalis. As regards the antibiograms, 96% of not VUR PNs responded to beta-lactams, 99% to aminoglycosides, and 80% to sulfonamides. For the VUR group, mean age was 3.0 years ± 3.0 years (range: 9 days of life–11 years) and mean number of episodes per patient was 2.0 ± 1.0 (range: 1–5); 83% of PNs were by Gram-negatives bacteria vs. 17% by Gram-positive: the most important Gram-negative bacteria were Pseudomonas aeruginosa (44%), Escherichia coli (27%), and Klebsiella spp. (12%), while Enterococcus spp. determined 90% of Gram-positive UTIs. Regimen ampicillin/ceftazidime (success rate: 72.0%) was compared to ampicillin/amikacin (success rate of 83.0%): no statistically significant difference was found (p = 0.09). The pathogens of PNs in uropathic patients are different from those of community-acquired PNs, and clinicians should be aware of their peculiar antibiotic susceptibility. An empiric therapy based on the association ampicillin + ceftazidime is therefore suggested.

Azide-Functionalized Derivatives of the Virulence-Associated Sugar Pseudaminic Acid: Chiral Pool Synthesis and Labeling of Bacteria.

Pseudaminic acid (Pse) is a significant prokaryotic monosaccharide found in important Gram‐negative and Gram‐positive bacteria. This unique sugar serves as a component of cell‐surface‐associated glycans or glycoproteins and is associated with their virulence. We report the synthesis of azidoacetamido‐functionalized Pse derivatives as part of a search for Pse‐derived metabolic labeling reagents. The synthesis was initiated with d‐glucose (Glc), which served as a cost‐effective chiral pool starting material. Key synthetic steps involve the conversion of C1 of Glc into the terminal methyl group of Pse, and inverting deoxyaminations at C3 and C5 of Glc followed by backbone elongation with a three‐carbon unit using the Barbier reaction. Metabolic labeling experiments revealed that, of the four Pse derivatives, ester‐protected C5 azidoacetamido‐Pse successfully labeled cells of Pse‐expressing Gram‐positive and Gram‐negative strains. No labeling was observed in cells of non‐Pse‐expressing strains. The ester‐protected and C5 azidoacetamido‐functionalized Pse is thus a useful reagent for the identification of bacteria expressing this unique virulence‐associated nonulosonic acid.

Impact of Polymyxin Resistance on Virulence and Fitness among Clinically Important Gram-Negative Bacteria

Humanity is facing an enormous and growing worldwide threat from the emergence of multi-drug-resistant (MDR) Gram-negative bacteria such as Escherichia coli , Klebsiella pneumoniae , and Acinetobacter baumannii . Polymyxin B and E (colistin) constitute the last-line therapies for treating MDR Gram-negative bacteria. Polymyxin is a cationic antibacterial peptide that can destroy the outer membrane of Gram-negative bacteria. With the increasing clinical application of polymyxin, however, there have been many reports of the occurrence of polymyxin-resistant Gram-negative bacteria. This resistance is mainly mediated by the modification or complete loss of lipopolysaccharide (LPS). LPS is also a virulence factor of Gram-negative bacteria, and alterations of LPS may correlate with virulence. Although it is generally believed that the biological costs associated with drug resistance may enable benign susceptible bacteria to overcome resistant bacteria when antibiotic pressure is reduced, some studies have shown that polymyxin-resistant bacteria are associated with higher virulence and greater fitness compared with their susceptible counterparts. To predict the development of polymyxin resistance and evaluate interventions for its mitigation, it is important to understand the relative biological cost of polymyxin resistance compared with susceptibility. The impact of polymyxin resistance mechanisms on the virulence and fitness of these three Gram-negative bacteria are summarized in this review.