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Klebsiella pneumoniae, a member of the ESKAPE pathogen group, is a prominent cause of hospital-acquired infections. The WHO has recognized carbapenem-resistant K. pneumoniae as a critical-one priority pathogen. These resilient superbugs have the ability to form biofilms and present a significant global threat. In the present study, we isolated and characterized a bacteriophage SAKp02, from hospital sewage, infectious to carbapenem-resistant K. pneumoniae patient isolates. SAKp02 could infect 43 of 72 clinical isolates, indicating a broad host spectrum. Whole genome analysis classified SAKp02 within the family Casjensviridae, with a 59,343 bp genome encoding 82 ORFs. Comparative genomic analysis revealed significant differences between SAKp02 and its closest viruses, indicating a distinct genetic makeup positioning it as a novel phage strain within the lineage. The SAKp02 genome comprises bacteriolytic enzymes, including holin, endolysin, and phage depolymerase, crucial for bacterial lysis and biofilm disruption. It reduced biofilm biomass by over threefold compared to the control and eradicated 99% of viable cells within a 4 h treatment period. Scanning electron microscopy corroborated the ability of the phage to dismantle biofilm matrices and lyse bacterial cells. Safe and effective treatments are warranted, and hence, the fully characterized lytic phages with therapeutic potential against drug-resistant clinical isolates of bacteria are needed. Our study is the first to report the antibacterial and antibiofilm activity of Casjensviridae phages, and our discovery of a novel K. pneumoniae phage broadens the arsenal against the bacteria. [ABSTRACT FROM AUTHOR]

Introduction: To ensure the appropriate usage of ceftazidime-avibactam (CAZ-AVI), recently introduced in our hospital, we aimed to determine susceptibility rates, enzyme analysis, and clonal relationship among strains, together with clinical data. Methodology: Between June 1 and September 30, 2021, demographic and microbiological data of the patients were recorded. In the obtained samples, meropenem and colistin minimal inhibitory concentration (MIC) levels, carbapenem resistance genes, and the clonal relationship were studied by molecular methods. CAZ-AVI was not used in any of the patients. Results: 140 carbapenem-resistant Klebsiella pneumoniae were isolated from 57 patients. Resistance to CAZ-AVI was found in 76 (54.3%) strains. Out of 57 patients, 31 (54.4%) isolates could be reached. Meropenem MIC level was ≥ 32 µg/mL in 26 (83.9%), and colistin MIC level was ≥ 4 µg/mL in 17 (54.8%) isolates. Enzyme analysis revealed NDM in 20 (64.5%), OXA-48 in 17 (54.8%), and KPC in seven (22.6%). NDM + OXA-48 was determined in 10 (32.2%) strains. NDM was determined in all CAZ-AVI resistant strains, OXA-48 in 16.1% (2/5) strains. Seven genotypes were detected. The largest cluster was genotype 3 clusters (11 isolates). Of 31 patients, 22 (71.0%) died. CAZ-AVI was susceptible in one of the patients who survived and four who died. Conclusion: Before using a new antibiotic, each center should determine the basal data and phenotypic/genotypic resistance ratios specific to that antibiotic. While a high NDM rate and low CAZ-AVI sensitivity limit the use of the drug in our center, it is clear that CAZ-AVI use in sensitive strains will decrease mortality. [ABSTRACT FROM AUTHOR]

Marine organisms are vast sources of bioactive components, with holothorians, or sea cucumbers being one of these sources. Therefore, sea cucumber specimens were examined as a promising source of natural substances that can be used as antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral, and antitumor agents. Holothuria papillifera was collected from Zaafarana, Ras Ghareb, and Altur stations and employed as a biological weapon against some pathogens. H. papillifera extracts showed high activity as antibacterial extracts against Staph. aureus and Enterococcus faecalis (Gram+ve), Escherichia coli, Salmonella typhi, and Klebsiella pneumoniae (Gram-ve). It also exhibited positive antifungal efficacy against Candida albicans and revealed moderate activity against liver cancer cells and a weak activity against breast cancer cells. Alternatively, H. papillifera crude extract has weak antiviral activity against CoxB4 and very weak antiviral efficacy against HAV (hepatitis A virus). Moreover, our results proved that H. papillifera extract had anti-inflammatory effects and exhibited antioxidant power. In conclusion, H. papillifera has broad antimicrobial activity against many pathogens, as well as antioxidant, antitumor, anti-inflammatory, among others. It is recommended to explain and characterize the composition of the bioactive compounds of H. papillifera to be included in the pharmaceutical industry. [ABSTRACT FROM AUTHOR]

Background and Objectives: Platelet concentrates (PC) are stored at 20–24°C to maintain platelet functionality, which may promote growth of contaminant bacteria. Alternatively, cold storage of PC limits bacterial growth; however, data related to proliferation of psychotrophic species in cold‐stored PC (CSP) are scarce, which is addressed in this study. Materials and Methods: Eight laboratories participated in this study with a pool/split approach. Two split PC units were spiked with ~25 colony forming units (CFU)/PC of Staphylococcus aureus, Klebsiella pneumoniae, Serratia liquefaciens, Pseudomonas fluorescens and Listeria monocytogenes. One unit was stored under agitation at 20–24°C/7 days while the second was stored at 1–6°C/no agitation for 21 days. PC were sampled periodically to determine bacterial loads. Five laboratories repeated the study with PC inoculated with lyophilized inocula (~30 CFU/mL) of S. aureus and K. pneumoniae. Results: All species proliferated in PC stored at 20–24°C, reaching concentrations of ≤109 CFU/mL by day 7. Psychrotrophic P. fluorescens and S. liquefaciens proliferated in CSP to ~106 CFU/mL and ~105 CFU/mL on days 10 and 17 of storage, respectively, followed by L. monocytogenes, which reached ~102 CFU/mL on day 21. S. aureus and K. pneumoniae did not grow in CSP. Conclusion: Psychrotrophic bacteria, which are relatively rare contaminants in PC, proliferated in CSP, with P. fluorescens reaching clinically significant levels (≥105 CFU/mL) before day 14 of storage. Cold storage reduces bacterial risk of PC to levels comparable with RBC units. Safety of CSP could be further improved by implementing bacterial detection systems or pathogen reduction technologies if storage is beyond 10 days. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae , a challenging opportunistic bacterium, became a notable global health concern owing to its clinical impact, widespread epidemiology and escalating antibiotic resistance. This comprehensive review delves into the multifaceted dimensions of K. pneumoniae , with a focus on its clinical implications, epidemiological patterns and the critical issue of antibiotic resistance. The review also emphasizes the implications of K. pneumoniae in the context of antimicrobial stewardship and infection control. Epidemiological aspects are scrutinized, shedding light on the global distribution and prevalence of K. pneumoniae. Factors influencing its transmission and persistence in healthcare facilities and communities are examined, with patient demographics, healthcare practices and geographical variations. The review centres on antibiotic resistance, a critical issue in the era of bacteria displaying resistance to multiple drugs. The mechanisms of resistance used by K. pneumoniae against various classes of antibiotics are elucidated, along with the alarming rise of carbapenem-resistant strains. It also highlights ongoing research efforts and innovative strategies aimed at addressing this critical public health issue. This comprehensive review offers a holistic understanding of K. pneumoniae , emphasizing its clinical significance, global epidemiology and the immediate necessity for effective strategies to combat antibiotic resistance. It serves as a valuable resource for healthcare practitioners, researchers and policymakers seeking to manage better and mitigate the impact of this pathogen on public health. [ABSTRACT FROM AUTHOR]

This document presents a study on the resistance mechanisms of Klebsiella pneumoniae to the antibiotic cefiderocol. The study focused on the overproduction of the SHV-1 β-lactamase in K. pneumoniae, which can reduce susceptibility to ceftazidime. The researchers sequenced and tested five clinical isolates of K. pneumoniae and found that those with SHV-1 overproduction exhibited resistance to cefiderocol. The study also identified a specific substitution in SHV-1 that enhanced its hydrolytic activity against cefiderocol. These findings contribute to our understanding of antibiotic resistance in K. pneumoniae and can help inform strategies to combat it. [Extracted from the article]

Objectives With the rise in antimicrobial resistance, there is a growing demand for rapid antimicrobial susceptibility testing (RAST). In this study, we applied the EUCAST RAST method to ESBL/carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolates without using advanced identification systems and analysed the effect of this method on mortality rates Also the clinical impact of this method on patients infected with these bacteria and its effect on mortality rates were investigated. Methods RAST was used for clinical blood cultures containing carbapenemase/ESBL-producing E. coli and K. pneumoniae without advanced identification systems (e.g. MALDI TOF), with preliminary identification by simple diagnostic tests (predicted RAST, or p-RAST), and its categorical agreement was investigated. The impact of the method on mortality was analysed by comparing the clinical data of patients whose blood cultures were subject to p-RAST (p-RAST group, n  = 49) and those who were not subject to p-RAST (non-RAST group, n  = 145). Results p-RAST results were analysed based on 539 antibiotic-bacteria combinations. Total error rates at 4, 6 and 8 h of incubation were 2.9%, 3.9% and 3.8%, respectively. In the p-RAST group, patients who did not receive appropriate antibiotics (29/45, 59.1%) were switched to appropriate treatment within 8 h at the latest. In contrast, in the non-RAST group, treatment of patients who received inappropriate antibiotics (79/145, 54.5%) could be changed after at least 24 h. Mortality rates were lower in the p-RAST group than in the non-RAST group (28.6% versus 51.7%, P  = 0.005). Conclusions p-RAST can be used safely in hospital laboratories with high rates of antimicrobial resistance and can reduce mortality rates by shortening the transition time to appropriate treatment. [ABSTRACT FROM AUTHOR]

Objectives Comprehensive data on the genomic epidemiology of hospital-associated Klebsiella pneumoniae in Ghana are scarce. This study investigated the genomic diversity, antimicrobial resistance patterns, and clonal relationships of 103 clinical K. pneumoniae isolates from five tertiary hospitals in Southern Ghana—predominantly from paediatric patients aged under 5 years (67/103; 65%), with the majority collected from urine (32/103; 31%) and blood (25/103; 24%) cultures. Methods We generated hybrid Nanopore–Illumina assemblies and employed Pathogenwatch for genotyping via Kaptive [capsular (K) locus and lipopolysaccharide (O) antigens] and Kleborate (antimicrobial resistance and hypervirulence) and determined clonal relationships using core-genome MLST (cgMLST). Results Of 44 distinct STs detected, ST133 was the most common, comprising 23% of isolates (n  = 23/103). KL116 (28/103; 27%) and O1 (66/103; 64%) were the most prevalent K-locus and O-antigen types. Single-linkage clustering highlighted the global spread of MDR clones such as ST15, ST307, ST17, ST11, ST101 and ST48, with minimal allele differences (1–5) from publicly available genomes worldwide. Conversely, 17 isolates constituted novel clonal groups and lacked close relatives among publicly available genomes, displaying unique genetic diversity within our study population. A significant proportion of isolates (88/103; 85%) carried resistance genes for ≥3 antibiotic classes, with the bla CTX-M-15 gene present in 78% (n  = 80/103). Carbapenem resistance, predominantly due to bla OXA-181 and bla NDM-1 genes, was found in 10% (n  = 10/103) of the isolates. Conclusions Our findings reveal a complex genomic landscape of K. pneumoniae in Southern Ghana, underscoring the critical need for ongoing genomic surveillance to manage the substantial burden of antimicrobial resistance. [ABSTRACT FROM AUTHOR]

This study evaluated the antioxidant, antidiabetic, anticholinesterase, anti-inflammatory, and antimicrobial effects of Algerian Cleome amblyocarpa through its chloroform fraction. Furthermore, phenolic, flavonoids and volatile compounds have been studied. The results indicated high phenols and flavonoids contents, and the gas chromatographymass spectrometry (GC-MS) analysis revealed the presence of various compounds, mainly 2- Tetracosane, Palmitic acid, 2- Aminothiazole, and Linolenic acid. Previous research supports the potential antidiabetic effects of palmitic acid and linolenic acid, while tetracosane has been studied for its anticholinesterase activity, validating the observed biological activities in our research. The fraction has demonstrated a substantial anti-inflammatory activity similar to the reference, antimicrobial actions against Escherichia coli, Klebsiella pneumonia, Bacillus cereus, Pseudomonas aeruginosa, and Candida albicans, as well as an antioxidant potential, and significant inhibition of key enzymes: alpha-amylase, alpha-glucosidase, acetyl, and butyrylcholinesterase (AChE and BChE), with interesting IC50 values. Our result found that Cléome amblyocarpa is a rich source of value-enhancing biocompounds, and it presents interesting antioxidant, antidiabetic, and anticholinesterase properties, as well as substantial anti-inflammatory and antimicrobial activities. These findings can be exploited in developing new formulations that may be useful in treating a variety of situations. [ABSTRACT FROM AUTHOR]

Objective To investigate clinical characteristics and risk factors of different stages of urinary tract infection after kidney transplantation. Methods Clinical data of 209 kidney transplant recipients were retrospectively analyzed. According to time points of postoperative follow-up, all recipients were divided into 3 stages: within 1 month post-kidney transplantation, 1-6 months post-kidney transplantation, and 7-12 months post-kidney transplantation. The incidence of urinary tract infection, urine culture results of recipients with urinary tract infection and drug resistance characteristics of common pathogens during different stages after kidney transplantation were analyzed. The strains of patients with recurrent urinary tract infection were identified. The risk factors of urinary tract infection and the effect of urinary tract infection on renal allograft function were analyzed. Results The urinary tract infection rate was 90.0% in the first stage, 49.3% in the second stage and 22.5% in the third stage. The urinary tract infection rates of male recipients undergoing living-related organ donation in the second and third stages were lower than those of female recipients (both P<0.05). Urine culture test yielded positive results in 60 cases, and 84 strains of pathogenic bacteria were detected, mainly Gram-negative bacteria, among which Klebsiella pneumoniae accounted for the highest proportion. Sixty-six recipients had recurrent urinary tract infection, and the detected pathogens included Klebsiella pneumoniae, Escherichia coli and Candida glabrata, etc. Univariate analysis showed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage. Preoperative urinary tract infection and donor type were the risk factors for urinary tract infection in the second stage. Gender and age of the recipients were the risk factors for urinary tract infection in the third stage. Multivariate analysis revealed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage. Gender and age of the recipients were the risk factors for urinary tract infection in the third stage (all P<0.05). In the third stage, 65 cases were cured and 38 cases were not cured. In the treated recipients, the serum creatinine level and white blood cell count were decreased after corresponding treatment than those before treatment (both P<0.05).Conclusions Gram-negative bacteria are the main pathogens of urinary tract infection in kidney transplant recipients, and drug resistance is relatively high. Postoperative use of antithymocyte globulin, female and old age are the risk factors for urinary tract infection in kidney transplant recipients. [ABSTRACT FROM AUTHOR]

Antimicrobial susceptibility patterns of bacterial pathogens isolated from patients with complicated urinary tract infections were analyzed using the national surveillance data, comprising 793 bacterial strains from eight clinically relevant species. Data were collected for the fourth national surveillance project from July 2020 to December 2021 by the Japanese Society of Chemotherapy, the Japanese Association for Infectious Disease, and the Japanese Society of Clinical Microbiology. Surveillance was supervised with the cooperation of 43 medical institutions throughout Japan. Fluoroquinolone required a minimum inhibitory concentration (MIC) of 2–64 mg/L to inhibit the 330 tested Escherichia coli strains. The proportion of levofloxacin-resistant E. coli strains increased from 28.6% in 2008 to 29.6% in 2011, 38.5% in 2015, and 44.5% in 2021. The proportion of levofloxacin-resistant strains of Pseudomonas aeruginosa also increased from previous survey results, showing a continuing downward trend. Conversely, the proportion of levofloxacin-resistant strains of Enterococcus faecalis decreased relative to previous reports. Neither multidrug-resistant P. aeruginosa nor carbapenem-resistant Enterobacteriaceae were detected. For methicillin-resistant Staphylococcus aureus (MRSA), the proportion of vancomycin-susceptible strains (MIC of 2 μg/mL) decreased from 14.7% to 7.7%. Bacterial strains that produced extended-spectrum β-lactamase included E. coli (82/330 strains, 24.8%), Klebsiella pneumoniae (11/68 strains, 16.2%), and Proteus mirabilis (4/26 strains, 15.4%). As compared to previous surveillance reports, these strains showed an increase in proportion over the years. [ABSTRACT FROM AUTHOR]

Limited experience exists with ceftazidime-avibactam (CAZ-AVI) in treating bacteremia caused by carbapenem-resistant Enterobacterales (CRE) and Pseudomonas aeruginosa (CRPA) in hematological patients. We performed a single-center, retrospective, observational study including patients who received CAZ-AVI for bacteremia due to CRE or CRPA between 2018 and 2022. The primary outcome was 30-day survival. We conducted a multivariable analysis to identify predictors of survival. 56 patients were included and 57 (41 CRE and 16 CRPA) strains were isolated. 35 strains produced carbapenemase, including 25 metallo-beta-lactamase (MBL) and 10 serine-beta-lactamase. 48 patients (85.7 %) received combination therapy. All patients with MBL-CRE bacteremia (n = 24) received combination therapy with aztreonam (AZT). The susceptibility rates to CAZ-AVI were only 26.8 % (11/41) in CRE and 80.0 % (8/10) in CRPA. The 30-day survival rates were 85.0 % (34/40) in the CRE group and 81.3 % (13/16) in the CRPA group. In patients with MBL-CRE bacteremia, the 30-day survival was as high as 91.7 % (22/24) due to combination with AZT. Ceftazidime did not influence the activity of aztreonam-avibactam against MBL-CRE in-vitro. Multivariable cox analysis revealed neutropenia >14 days (P = 0.002, HR: 34.483, 95%CI: 3.846–333.333) and a higher Pitt bacteremia score (P = 0.005, HR: 2.074, 95%CI: 1.253–3.436) were risk factors for 30-day survival. CAZ-AVI is highly effective in treating bacteremia due to CRPA and serine-beta-lactamase CRE. The combination of avibactam with AZT is highly effective in treating bacteremia due to AZT-resistant MBL producers. [ABSTRACT FROM AUTHOR]

To assess the effectiveness of colistin (administered as colistimethate sodium-CMS) and polymyxin B (PMB) for the treatment of bloodstream infections (BSIs) caused by carbapenem-resistant Klebsiella pneumoniae (CRKP). This retrospective cohort included hospitalized adult patients with CRKP BSIs from a single tertiary-care hospital. A univariate analysis comparing CMS and PMB groups was carried out and an inverse-probability propensity score (IPPS) was created. An IPPS-adjusted Cox regression model for 30-day mortality was performed including covariates potentially associated with mortality. A total of 100 patients with CRKP BSI (87 were KPC-producing isolates) were included. The 30-day mortality was 42.0 %:17/46 (38.8 %) and 25/54 (44.6 %) patients of CMS and PMB groups, respectively, P = 0.54 (incidence rate, 18.9 and 21.7/1000 patients-day in CMS and PMB groups, respectively, P = 0.62). No statistically significant difference in 30-day mortality rate was observed in a model adjusted for Pitt bacteremia score, high-risk primary site and IPPS, which included age, intensive care unit admission, minimal inhibitory concentration, previous colonization by CRKP, diabetes mellitus, malignancy, neutropenia, meropenem use before BSI, adjuvant therapy with meropenem and amikacin, and time to start polymyxin. Acute kidney injury (AKI) occurred in 52.0 % of patients, with no significant differences between groups (47.8 % and 57.4 % for CMS and PMB, respectively, P = 0.83). In-hospital mortality was 47,7 % and 50.0 % in CMS and PMB groups, respectively, P = 0.82. There was no difference in 30-day mortality and AKI rates among patients with CRKP BSI treated with PMB or CMS. [ABSTRACT FROM AUTHOR]

Introduction: In patients with suspected pneumonia who are tested with respiratory culture and multiplex PCR, the potential added benefit of next-generation sequencing technologies is unknown. Methods: This was a single-center, retrospective study in which residual bronchoalveolar lavage (BAL) specimens were retrieved from hospitalized patients. We compared its research-use-only Respiratory Pathogen Illumina Panel (RPIP) results to culture and BioFire® FilmArray Pneumonia Panel (BioFire® PN) results from critically ill patients. Results: In total, 47 BAL specimens from 47 unique patients were included. All BAL samples were tested with culture and multiplex PCR. In total, 38 of the 47 BALs were consistent with a clinical picture of pneumonia per chart review. Additional testing of the 38 samples with the RPIP identified a new bacterium in 20 patients, a new virus in 4 patients, a new bacterium plus virus in 4 patients, and no additional organisms in 10 patients. In 17 (44.5%) of these patients, the RPIP results could have indicated an antibiotic addition. Compared with cultures, the RPIP had an overall sensitivity of 64% and specificity of 98%, with a 0% sensitivity for fungus and 14% sensitivity for mycobacteria. Compared with BioFire® PN, the RPIP was 70% sensitive and 99% specific, with a 74% sensitivity for bacteria and 33% sensitivity for viruses. The RPIP was 29% more sensitive for HAP/VAP bacterial targets compared with CAP. Conclusions: Emerging NGS technologies such as the RPIP may have a role in identifying the etiology of pneumonia, even when patients have BAL culture and multiplex PCR results available. Similar to prior studies evaluating RPIP, our study showed this platform lacked sensitivity when compared with cultures, particularly for fungi and mycobacteria. However, the high specificity of the test can be leveraged when clinicians are seeking to rule out certain infections. [ABSTRACT FROM AUTHOR]

Introduction: Hypervirulent Klebsiella pneumoniae (hvKp) has emerged as a pathogen of global concern that is hypermucoviscous, causing infections and rapidly acquiring Antimicrobial Resistance (AMR). HvKp is more virulent than classical K. pneumoniae (cKp) and frequently infects healthy individuals with community-acquired illnesses. Since hvKp commonly exists in the gastrointestinal tract, its communicable spread affects the general public and healthcare facilities. The study was necessary due to the rapid evolution of hvKp, the need to identify novel, reliable agents of hypervirulence and drug resistance, and more reliable targets for therapeutic intervention. Aim: To investigate the prevalence of hvKp strains (rmpA and magA genes) and drug-resistant patterns in clinical isolates of Klebsiella pneumoniae (K. pneumoniae) in a tertiary care hospital in Puducherry, India. Materials and Methods: The cross-sectional study included a total of 100 non duplicate consecutive isolates of K. pneumoniae collected from Mahatma Gandhi Medical College and Research Institute and Hospital, Puducherry, India. These isolates were recovered from various clinical specimens such as urine, sputum, pus, blood, and other miscellaneous specimens mainly obtained from both outpatients and inpatients between August 2021 and April 2022. The antibiotic susceptibility of cultured K. pneumoniae was determined by the disk-diffusion test following the 2020 Clinical and Laboratory Standards Institute (CLSI) guidelines. The rmpA and magA genes were detected using conventional Polymerase Chain Reaction (PCR). PCR products were sequenced using Sanger sequencing. Statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) statistics software for Windows, version 15.0 (available from SPSS Inc., Chicago, USA). A p-value <0.05 was considered statistically significant. Results: A total of 100 isolates were studied for phenotypic and genotypic properties. HvKp was identified phenotypically using the string test, and further, the rmpA and magA genes were identified genotypically using PCR. Out of 100 isolates, the phenotypic study showed 11 (11%) isolates as string test positive. Furthermore, genotypic results showed that 22 (22%) isolates expressed the rmpA gene, 11 (11%) isolates expressed the magA gene, and 7 (7%) isolates expressed both rmpA and magA. Most of the hvKp isolates were obtained from urine specimens of K. pneumoniae isolates and showed more resistance than cKp towards all classes of antibiotics. Conclusion: The high prevalence of rmpA and magA genes suggests their strong role and capability as genetic markers for the identification of hvKp in the laboratory. This study indicated the important role of genetic elements in the emergence of drug resistance in hypervirulent strains and showed that hvKp has a multidrug resistance pattern. [ABSTRACT FROM AUTHOR]

Objective: We explored whether the Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification (R-M) systems are compatible and act together to resist plasmid attacks. Methods: 932 global whole-genome sequences from GenBank, and 459 K. pneumoniae isolates from six provinces of China, were collected to investigate the co-distribution of CRISPR-Cas, R-M systems, and blaKPC plasmid. Conjugation and transformation assays were applied to explore the anti-plasmid function of CRISPR and R-M systems. Results: We found a significant inverse correlation between the presence of CRISPR and R-M systems and blaKPC plasmids in K. pneumoniae, especially when both systems cohabited in one host. The multiple matched recognition sequences of both systems in blaKPC-IncF plasmids (97%) revealed that they were good targets for both systems. Furthermore, the results of conjugation assay demonstrated that CRISPR-Cas and R-M systems in K. pneumoniae could effectively hinder blaKPC plasmid invasion. Notably, CRISPR-Cas and R-M worked together to confer a 4-log reduction in the acquisition of blaKPC plasmid in conjugative events, exhibiting robust synergistic anti-plasmid immunity. Conclusions: Our results indicate the synergistic role of CRISPR and R-M in regulating horizontal gene transfer in K. pneumoniae and rationalize the development of antimicrobial strategies that capitalize on the immunocompromised status of KPC-KP. [ABSTRACT FROM AUTHOR]

Background: Infections due to Citrobacter species are increasingly observed in hospitalized patients and are often multidrug-resistant. Yet, the magnitude and burden of Citrobacter spp. resistance in the hospital setting have not been reported. We aimed to evaluate the epidemiology of Citrobacter spp. infections among hospitalized patients, their main resistance patterns and Citrobacter spp. involvement in hospital outbreaks. Methods: We conducted a systematic review and meta-analysis of published literature (PROSPERO registration Jan-2023, CRD42023390084). We searched Embase, Medline and grey literature for studies on hospitalized patients diagnosed with Citrobacter spp. infections, and nosocomial outbreaks due to Citrobacter spp. published during the years 2000–2022. We included observational, interventional, surveillance studies and outbreak reports. Outcomes of interest were the frequency of Citrobacter spp. infections among hospitalized patients and 3rd generation cephalosporin and/or carbapenem resistance percentages in these infections. We used random-effects models to generate pooled outcome estimates and evaluated risk of bias and quality of reporting of outbreaks. Results: We screened 1609 deduplicated publications, assessed 148 full-texts, and included 41 studies (15 observational, 13 surveillance and 13 outbreak studies). Citrobacter spp. urinary tract- and bloodstream infections were most frequently reported, with Citrobacter freundii being the main causative species. Hospital-acquired infection occurred in 85% (838/990) of hospitalized patients with Citrobacter infection. After 2010, an increasing number of patients with Citrobacter spp. infections was reported in observational studies. Pooled frequency estimates for Citrobacter spp. infections could not be generated due to lack of data. The pooled prevalence of ESBL and carbapenemase producers among Citrobacter isolates were 22% (95%CI 4–50%, 7 studies) and 18% (95%CI 0–63%, 4 studies), respectively. An increased frequency of reported Citrobacter outbreaks was observed after 2016, with an infection/colonization ratio of 1:3 and a case-fatality ratio of 7% (6/89 patients). Common outbreak sources were sinks, toilets, contaminated food and injection material. Implemented preventive measures included environmental cleaning, isolation of positive patients and reinforcement of hand hygiene. Only seven out of 13 outbreaks (54%) were definitively controlled. Conclusion: This review highlights the clinical importance of endemic and epidemic Citrobacter spp. in healthcare settings. As an emerging, multidrug‑resistant nosocomial pathogen it requires heightened awareness and further dedicated surveillance efforts. [ABSTRACT FROM AUTHOR]

Otitis media is frequently prevalent in developing countries and is primarily caused by bacteria. The current study aimed to study the patients to assess the occurrence rate, susceptibility pattern, and prevalence rate of metallo beta-lactamase (MBL) producing gram-negative bacteria to prevent the prevalent strains and direct the physicians to appropriate antibiotic therapy. The research study was conducted over nine-months, from November 2020 to July 2021, in which a total of 110 samples were collected from the outdoor-patients’ ear, nose, and throat (ENT) wards using sterile swabs and inoculated immediately on suitable culture media. The isolates were identified via conventional biochemical tests, and the susceptibility pattern of each isolate was tested using the Kirby-Bauer disc diffusion method. Out of the 110 total samples, merely 88.18% (97/110) ear samples were detected positive for the various strains of bacteria both in male and female patients with a mean age of 18.21 ± 15.86, contributing 54.64% and 45.36%, respectively. The most prevalent bacterial strain was Staphylococcus aureus (49.48%), followed by Pseudomonas aeruginosa (16.50%), Proteus spp. (13.40%), S. epidermidis (7.22%), E. coli (6.19%), S. saprophyticus (4.12%), and Klebsiella spp. (3.09%). Of the gram-negative isolated strains, only 18.75% (3/16) of the P. aeruginosa strains exhibited MBL production. A high prevalence rate was observed in the age-group 0–4 years (19.58%), followed by the age-groups, 5–9 and 15–19 (16.49% each), and the age-group >39 (10.31%). Levofloxacin, ciprofloxacin, amikacin, and fusidic acid were highly effective against the gram-positive, whereas ciprofloxacin, amikacin, and levofloxacin were potent against the gramnegative (except P. aeruginosa), while carbenicillin, meropenem and imipenem against the P. aeruginosa. The MBL-producers were highly (100%) resistant to IMP and MEM while highly (100%) sensitive to CAZ. In conclusion, the current study showed a high prevalence of S. aureus, P. aeruginosa, and MBL-producing P. aeruginosa (18.75%). Nevertheless, a high degree of resistance to antibiotics was noted, which could not be ignored. [ABSTRACT FROM AUTHOR]

Key Clinical Message: Infections in infantile hemangiomas (IHs) are generally limited, and only few cases have been reported. The rapid expansion of an ulcerated IH should raise concern for possible complications to monitor and provide immediate therapeutic interventions. This case highlights the importance of prompt treatment in large segmental IHs to prevent ulceration and related complications, including bleeding and superinfection. IH is a benign tumor proliferating during early infancy. While many IHs spontaneously resolve, complications like ulceration, bleeding, and potential damage to vital organs can occur, leading to pain, infection, and scarring. A 6‐month‐old girl with a previously treated IH on her left leg developed a Klebsiella‐infected ulcer at the site. The ulcer resulted from non‐standard treatments used before admission. Upon hospitalization, she was initially treated with cefepime and propranolol, but a week later, the wound culture revealed Klebsiella pneumoniae, prompting a switch to piperacillin/tazobactam. After successfully managing the infection and bleeding, the child was discharged in good condition with orders to continue treatment with propranolol for at least a year. This case highlights the potential of IHs to become infected even with uncommon germs such as Klebsiella and the importance of receiving appropriate medical care to prevent further complications. [ABSTRACT FROM AUTHOR]

Multidrug resistance is a global health problem that is closely related to microbial resistance. Klebsiella pneumonia and Proteus mirabilis are opportunistic bacteria that cause urinary tract infections, especially in the use of catheters in hospitals in inpatient care. Handling it using commercial antibiotics can increase the effect of antibacterial resistance if the use is irrational and neglected. Avocado leaves have been reported to have antibacterial, antidiabetic and antioxidant activity. The purpose of this study was to observe into the antibacterial activity of avocado leaves ethanol extract against K. pneumoniae and P. mirabilis. The minimum inhibitory concentration (MIC) was determined using the Kirby-Bauer method, the minimum bactericidal concentration (MBC) was determined using the streaking method of the inhibition zone formed, and the mechanism of bacterial membrane leakage cell was measured using UVVIS spectrophotometry at wavelengths of 260 and 280 nm. The MIC value was found to be 3.125 mg/mL in both bacteria, with an inhibition zone of 6.43 ± 0.06 mm (K. pneumoniae) and 6.73 ± 0.15 mm (P. mirabilis). The MBC value at 200 mg/mL with a percent reduction of 98.35 % (K. pneumoniae) and at 100 mg/mL with a percent reduction of 98.06 % (P. mirabilis). The supernatant of a membrane leakage cell (DNA and protein) was absorbed at wavelengths of 260 and 280 nm. It can be concluded that the ethanolic extract of avocado leaves has antibacterial potential against clinical isolates of K. pneumoniae and P. mirabilis. [ABSTRACT FROM AUTHOR]

Background: People living with HIV/AIDS (PLWHA) are prone to opportunistic bacterial infections caused by multidrug-resistant organisms. The aim of this study was to determine the susceptibility of Enterobacterales isolated from urine and stool samples of PLWA attending the BOYAMBI Hospital Center, Kinshasa, Democratic Republic of the Congo to commonly used antibiotics and to detect extended-spectrum β-lactamases (ESBLs) producers among the isolates. Methodology: A total of 163 HIV-infected patients attending the BOYAMBI Hospital, Kinshasa, DRC, were randomly selected for this study. Urine samples were collected from 108 patients, while stool samples were collected from 55 of them. Samples were cultured in MacConkey agar and identified using conventional microbiological methods. Antibiotic susceptibility on each isolate to selected antibiotics was performed by the disc diffusion method. Phenotypic detection of ESBL was done by the double-disc synergy test. Results: A total of 120 Enterobacterales were isolated from the samples of the 163 HIV-infected patients with 65 of 108 (60.2%) urine and 55 of 55 (100.0%) stool samples. Escherichia coli was the most frequent bacterial species from both urine and stool with 84 (70.0%), followed by Klebsiellaspecieswith23 (19.2%). Other bacterial pathogens were Citrobacter (n=6, 5.0%), Enterobacter (n=4, 3.3%), Proteus (n=2, 1.7%) and Morganella (n=1, 0.8%) species. Escherichia coli isolates were resistant to amoxicillin (90.5%), sulfamethoxazole-trimethoprim (81.0%), ciprofloxacin (77.4%), ceftriaxone (77.4%), ceftazidime (73.8%), amoxicillin–clavulanic acid (61.9%), imipenem (60.7%), and cefotaxime (50.0%). Klebsiella pneumoniae isolates were resistant to ceftazidime (95.7%), ceftriaxone (91.3%), imipenem (91.3%), ciprofloxacin (87%), sulfamethoxazole-trimethoprim (78.3%), cefotaxime (56.5%), and amoxicillin–clavulanic acid (52.2%). Citrobacter, Enterobacter, Morganella and Proteus species were resistant to the majority of antibiotics. The rate of ESBL production was 23.0% (28/120) with Citrobacter spp being the most frequent ESBL-producer, followed Klebsiella spp, Enterobacter spp and E. coli. Conclusion: The results obtained showed a high rate of ESBL-producing Enterobacterales isolates which were multi-drug resistant. Nitrofurantoin, gentamicin, chloramphenicol, cefixime and nalidixic acid were the most active antibiotics against the isolates. [ABSTRACT FROM AUTHOR]

Ninty-four clinical samples were collected as swabs from cows suffered from different types of wounds in Baghdad governorate. Samples were cultured on different media and biochemically tested for identify the type of bacteria that colonize in the wounds and to isolate and identify P. aeruginosa. The results revealed that out of 94 samples collected from wounds, 87 bacteria isolate (90.55%) were grow bacterial growth and consider positive samples, while negative growth appear to be 7.4%. The profile of bacteria in the cultured samples revealed bacteria as fellow: P. aeruginosa 36 (36.78%) was the most common isolate followed by S. aureus 22(25.28%), E. coli 21(24.1%), K. pneumoniae 5 (5.7%), P. mirabilis 3(3.4%). Thirty six P. aeruginosa isolates were tested for antibiotic sensitivity. All isolate (100.0%) are resistant to Ampicilin-Sulbactam, Ticracillin-pyostacine Ticracillin-,clavulnic acid. and Ticracillin, these isolates show high resistance to Cotrimoxazol 94.4%, Cefepim 77.7%, Ceftazidime 66.6%, Imipenem 63.8%, Meropenem 61.1% while show low resistance to Gentamicin 44.4%, Tobramycin 38.8%, Piperacillin-pyostacine 27.7%, Piperacillin-Tazobactam 30.5%, Piperacillin- Tazobactam-pyostacine 33.3%, Colistin 33.3%, Ciprofloxacin 22.2% and Amikacin 22.2%, and among these antibiotics, Amikacin and Ciprofloxacin was the most effective antibiotic against P. aeruginosa isolates. [ABSTRACT FROM AUTHOR]

BACKGROUND: K.pneumoniae is a gram-negative bacterium belonging to the Enterobacteriaceae family. This microorganism is a part of the healthy microbiome of individuals and colonizes many parts of the body. The incidence of carbapenemresistant Klebsiella pneumoniae (CRKP) infections is increasing globally. In India, around 65% of K. pneumoniae isolates are resistant to carbapenem antibiotics. The blaNDM is the predominant carbapenem-resistant gene in CRKP isolates. AIM AND OBJECTIVE: To Study the Molecular Characterization of blaNDM-1 Resistant genes in Carbapenem Resistance Klebsiella Pneumonia Clinical isolates. MATERIAL AND METHODS:This was a cross sectional study carried out for a period of 1 year i.e, during 2023 to 2024at K.G.M.U, Lucknow . Samples like urine, sputum, ET tube, pleural fluid, pus, CSF, blood and Ascitic fluid were included in this study. Carbapenemase resistance was detected phenotypically by MHT, mCIM and eCIM methods and genotypically by using PCR where the DNA extraction was done and the resistant gene blaNDM-1 was confirmed by PCR assay. RESULT:A total of 350 isolates of K. pneumoniae were collected from patients, with 127(36.28%) identified as CRKP. There were 30 (57.69%) isolates obtained from male and 22(42.30%) were obtained from female, while the maximum number of patients belong to the age group <50 years and maximum patients were engaged as Student/Housewife/Retired (44.09%) followed by Clerical/Shopkeeper/Farmer (36.22%) and others. In our study,it was also observed that, in phenotypic test 81.88% were positive for M-CIM while 77.95% were positive for E-CIM and in genotypic test there were 86.61 % positive for NDM-1. CONCLUSION: The study highlighted the existence of carbapenemase-producing K. pneumoniae, particularly in blaNDM-1, in patients with comorbidities. Our findings emphasize the importance of the molecular characterization of resistance-determinant-carrying bacterial pathogens as a part of infection control and prevention in hospital settings. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae can enter a viable but nonculturable (VBNC) state to survive in unfavorable environments. Our research found that high-, medium-, and low-alcohol-producing K. pneumoniae strains are associated with nonalcoholic fatty liver disease. However, the presence of the three Kpn strains has not been reported in the VBNC state or during resuscitation. In this study, the effects of different strains, salt concentrations, oxygen concentrations, temperatures, and nutrients in K. pneumoniae VBNC state were evaluated. The results showed that high-alcohol-producing K. pneumoniae induced a slower VBNC state than medium-alcohol-producing K. pneumoniae, and low-alcohol-producing K. pneumoniae. A high-salt concentration and microoxygen environment accelerated the loss of culturability. Simultaneously, both real-time quantitative PCR and droplet digital PCR were developed to compare the quantitative comparison of three Kpn strain VBNC states by counting single-copy gene numbers. At 22°C or 37°C, the number of culturable cells decreased significantly from about 108 to 105-106 CFU/mL. In addition, imipenem, ciprofloxacin, polymyxin, and phiW14 inhibited cell resuscitation but could not kill VBNC-state cells. These results revealed that the different environments evaluated play different roles in the VBNC induction process, and new effective strategies for eliminating VBNC-state cells need to be further studied. These findings provide a better understanding of VBNC-state occurrence, maintenance, detection, and absolute quantification, as well as metabolic studies of resuscitation resistance and ethanol production. [ABSTRACT FROM AUTHOR]

Background: Escherichia coli and Klebsiella pneumoniae are two important commensals with pathogenic potentiality in animals and birds, causing intestinal and extra-intestinal infections. These are somewhat zoonotic too via different routes. So, the present study was undertaken to investigate the prevalence of these two bacteria from cloaca and environment of ducks from different districts of West Bengal, followed by detailed characterization and antibiogram. Methods: Around 197 duck and environmental samples were collected from different districts of West Bengal. E. coli and K. pneumoniae were isolated from collected samples as per standard methods followed by serotyping of the E. coli isolates. The detection of ESBL/ACBL producing isolates was done both phenotypically and genotypically by detecting the presence of antibiotic-resistant genes. Virulence property and biofilm-producing ability of these isolates were also studied. Antibiogram of the ESBL and biofilm positive isolates were done using common 12 antibiotics by disc diffusion method. Result: A total of 134 (68.02%) Escherichia coli and 33 (16.75%) Klebsiella pneumoniae isolates were recovered and confirmed by PCR. The prevalence of E. coli and K. pneumoniae in cloacal swabs were less in comparison to the environmental samples (72.09% and 20.93%). Approx. 79.85% E. coli and 72.73% K. pneumoniae isolates were found to be the ESBL/ACBL producers in vitro. E. coli and K. pneumoniae isolates possessed all major antibiotic-resistant genes with the blaAmpC (64.18% and 48.48%) being the most prevalent one. Twenty three (17.16%) E. coli isolates were found positive for the virulence genes, of which stx1 (39.13%) was the most common one as detected by multiplex PCR. No virulence gene was found in K. pneumoniae isolates. Ninety one (67.91%) E. coli and 26 (78.79%) K. pneumoniae isolates possessed the biofilm-producing genes. All the ESBL and biofilms producing E. coli and K. pneumoniae isolates showed 100% - 95.24% resistance to ampicillin. A high percentage of resistances were also seen against cefotaxime, ticarcillin/ clavulanic acid, ertapenem, ceftazidime, norfloxacin and ciprofloxacin, which may be due to several reasons. Maximum overall sensitivity was detected against chloramphenicol, co-trimoxazole and gentamicin. [ABSTRACT FROM AUTHOR]

Background: This study focuses on the AMR profiles in E. coli isolated from captive mammals at EcoZoo San Martín, Baños de Agua Santa, Ecuador, highlighting the role of wildlife as reservoirs of resistant bacteria. Aims: The aim of this research is to investigate the antimicrobial resistance profiles of E. coli strains isolated from various species of captive mammals, emphasizing the potential zoonotic risks and the necessity for integrated AMR management strategies. Materials & Methods: A total of 189 fecal samples were collected from 70 mammals across 27 species. These samples were screened for E. coli, resulting in 90 identified strains. The resistance profiles of these strains to 16 antibiotics, including 10 β‐lactams and 6 non‐β‐lactams, were determined using the disk diffusion method. Additionally, the presence of Extended‐Spectrum Beta‐Lactamase (ESBL) genes and other resistance genes was analyzed using PCR. Results: Significant resistance was observed, with 52.22% of isolates resistant to ampicillin, 42.22% to ceftriaxone and cefuroxime, and 27.78% identified as ESBL‐producing E. coli. Multiresistance (resistance to more than three antibiotic groups) was found in 35.56% of isolates. Carnivorous and omnivorous animals, particularly those with prior antibiotic treatments, were more likely to harbor resistant strains. Discussion: These findings underscore the role of captive mammals as indicators of environmental AMR. The high prevalence of resistant E. coli in these animals suggests that zoos could be significant reservoirs for the spread of antibiotic‐resistant bacteria. The results align with other studies showing that diet and antibiotic treatment history influence resistance profiles. Conclusion: The study highlights the need for an integrated approach involving veterinary care, habitat management, and public awareness to prevent captive wildlife from becoming reservoirs of antibiotic‐resistant bacteria. Improved waste management practices and responsible antibiotic use are crucial to mitigate the risks of AMR in zoo environments and reduce zoonotic threats. [ABSTRACT FROM AUTHOR]

Background: The escalation of antimicrobial resistance (AMR) in recent years has been of major public health concern globally. Escherichia coli are amongst the bacteria that have been targeted for AMR surveillance due to their ability to cause infection in both animals and humans. Their propensity to produce extended spectrum beta‐lactamases further complicates the choices of treatment regimens. Objectives: To investigate the prevalence of antimicrobial‐resistance in E. coli strains isolated from faecal samples of dogs and cats from selected veterinary surgeries and animal shelters from Harare, Zimbabwe. Materials and Methods: A cross‐sectional study was carried out to select animals by a systematic random procedure. Faecal samples were collected for culture and isolation of E. coli. Their susceptibility to antimicrobial drugs was assessed using the disc diffusion method. Results: A total of 95% (133/140) of the samples from cats (n = 40) and dogs (n = 93) yielded E. coli. Resistance was recorded for ampicillin (45.9%), trimethoprim‐sulphamethoxazole (44.4%), nalidixic acid (29.3%), ceftazidime (15.8%) and azithromycin (12.8%), but not for gentamicin and imipenem. A total of 18% of the isolates were multi‐drug‐resistant where resistance to nalidixic acid, ampicillin and trimethoprim‐sulphamethoxazole predominated. Conclusion: We observed relatively high AMR of E. coli strains against ampicillin. The isolation of multi‐drug‐resistant strains of E. coli may signal the dissemination of resistance genes in the ecosystem of these bacteria which may have a public health impact. [ABSTRACT FROM AUTHOR]

Introduction: Urinary tract infections (UTIs) caused by antimicrobial-resistant Enterobacterales are a global health threat. There are limited surveillance data available to characterize the prevalence of antimicrobial resistance among outpatients in the United States (US). Methods: This retrospective cohort (database) study investigated co-resistance among Escherichia coli and Klebsiella pneumoniae urinary isolates from US female outpatients aged ≥ 12 years with presumed uncomplicated UTI (uUTI), ≥ 3 months of data (2011–2019), and antimicrobial susceptibility testing results. Eligible isolates were the first urinary E. coli or K. pneumoniae isolate per patient collected within 30 days; classified as not susceptible (NS) if antimicrobial susceptibility testing results were intermediate or resistant to each antibiotic tested. Four resistance phenotypes were identified: NS to fluoroquinolones (FQ), trimethoprim/sulfamethoxazole (SXT), nitrofurantoin (NTF), and extended-spectrum β-lactamase+/third-generation cephalosporin (ESBL+/3GC NS). Co-resistance phenotypes included all possible combinations of resistance to ≥ 2 drug classes. Results: Of 1,513,882 E. coli isolates and 250,719 K. pneumoniae isolates, 856,918 and 187,459 isolates with ≥ 1 resistance phenotype were included in the analysis, respectively. The most common resistance phenotypes were SXT NS for the E. coli isolates (44.8%) and NTF NS for the K. pneumoniae isolates (75.5%), while ESBL+/3GC NS comprised 11.2 and 5.9%, respectively. Among ESBL+/3GC NS E. coli isolates, 72.4, 56.7, and 46.6% were co-resistant to FQ, SXT, and FQ + SXT, respectively. For ESBL+/3GC NS K. pneumoniae isolates, 65.7 and 45.7% were co-resistant to SXT and FQ + SXT. Conclusion: Both species exhibited high rates of co-resistance, emphasizing the need to raise awareness of co-resistance and of the unmet need for effective treatment options for uUTI. [ABSTRACT FROM AUTHOR]

Introduction: This study aimed to determine the pathogen distribution and drug susceptibility of diabetic foot wound secretions in a tertiary hospital in a coastal area of southeastern China to guide clinical antibiotic selection. Methods: A retrospective analysis was conducted on 212 patients with diabetic foot hospitalized at Xiamen Third Hospital from 2018 to 2023, and foot wound secretions were collected for microbial culture and drug susceptibility testing. Results: Among 212 cases of patients with diabetic foot wound secretions, 163 cases (76.9%) were cultured with pathogenic bacteria, and a total of 207 strains of pathogenic bacteria were cultured, including 75 strains (36.23%) of Gram-positive (G+) bacteria, 118 strains of Gram-negative (G−) bacteria (57.00%), 14 strains of fungi (6.76%), 120 cases of single microorganism infection (73.62%), 43 cases of mixed infection (26.38%), and 15 strains of multidrug-resistant bacteria (7.25%). The top three pathogenic bacteria were Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. G+ bacteria were dominated by S. aureus. Drug susceptibility results showed that G+ bacteria were highly susceptible to vancomycin, linezolid, tigecycline, quinupristin/dalfopristin, rifampicin, and furotoxin, and somewhat resistant to penicillin, erythromycin, clindamycin, and cefoxitin. Among G− bacterial infections, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Proteus were the major species. Drug susceptibility testing indicated that carbapenems such as imipenem and ertapenem were the most effective antibacterial drugs against G− strains, followed by amikacin, piperacillin, and tazabactams to which these bacteria were also relatively sensitive, while resistance to penicillins and first-generation cephalosporins increased significantly. We isolated one strain of pathogenic bacteria from a Wagner grade 1 ulcer, which was G+ bacteria. In Wagner grade 2 ulcers, the distribution of pathogenic bacteria was mainly G+ bacteria. In Wagner grade 3 and 4 ulcers, the distribution of pathogenic bacteria was mainly G− bacteria, and the increased rate of mixed infection was mainly due to mixed infection of G+ and G−. Two strains of pathogenic bacteria were isolated at Wagner grade 5, which were mixed infections of G+ and G−. Conclusions: Pathogenic bacteria in diabetic foot wounds are predominantly G− bacteria, followed by G+ bacteria. As the Wagner ulcer grade increases, the distribution of pathogenic bacteria changes from G+ bacteria to G− bacteria, and the mixed infection rate increases. G+ bacteria are highly susceptible to vancomycin, linezolid, tigecycline, quinupristin/dalfopristin, rifampicin, and furotoxin, and somewhat resistant to penicillin, erythromycin, clindamycin, and cefoxitin. G− bacteria are more sensitive to the antimicrobial drugs ertapenem, imipenem, amikacin, piperacillin tazobactam, and have high resistance to penicillin and first-generation cephalosporins. [ABSTRACT FROM AUTHOR]

The emergence of extensively-resistant strains of Klebsiella pneumoniae (K. pneumoniae) in healthcare settings is linked to prolonged hospitalization and uncontrolled use of antibiotics. There is a paucity of data regarding the prevalence and mechanisms of colistin and fosfomycin resistance encoding genes rate and mechanisms in Iran. The objective of this study was to determine the prevalence of biofilm formation and fosfomycin and colistin resistance among K. pneumoniae strains producing ESBL and carbapenemases by detecting the mcr-1, mcr-2, and fosA genes in Tehran, Iran, during the 2020-2021 period. After collecting 73 samples, the isolates were identified using biochemical tests. Antibiotic susceptibility test was performed using the disk diffusion method. The phenotypic determination of extended-spectrum beta-lactamases (ESBLs) and carbapenemase enzymes was conducted using combined disk and CARBA-NP tests, respectively. The biofilm formation was conducted using a microtiter tissue plate assay. Polymerase chain reaction (PCR) was employed to detect the mcr-1, mcr-2 and fosA genes, which are associated with colistin and fosfomycin resistance, respectively. The highest resistance rate was observed against ampicillin (97%), chloramphenicol (90%), and ciprofloxacin (87%), respectively. In contrast, the lowest resistance rate was noted against gentamicin (4%), amikacin (10%), and cotrimoxazole (18%). Moreover, 44 and 23 isolates were identified as ESBL and carbapenemase -producing K. pneumonia), respectively. Of the fortyeight isolates that formed strong biofilms, one was a non-biofilm producer. The PCR test revealed the amplification of the fosA2 gene in four isolates and the mcr-2 genes in one isolate. However, no amplification of the fosA3 or mcr-1 genes was observed. The present study demonstrated that the frequency of K. pneumoniae isolates producing ESBL and carbapenemase, as well as mcr-1, mcr-2 and fosA genes, was relatively low. However, given the potential for these genes to be disseminated more widely, it is imperative to implement effective isolation and control measures. Moreover, these strains demonstrated the capacity to form biofilms in vitro, which can lead to persistent infections in the hospital settings. [ABSTRACT FROM AUTHOR]

Background: Having a weakened immune system can make patients easily get nosocomial infection (NI) with multi-drug resistant (MDR) bacteria and put them in a dangerous situation. It causes long hospital stays, disability, economic burden, and even death. The present study aimed to determine the prevalence of NI in patients suffering from COVID-19. Methods: In this retrospective study, the information on 250 patients suffering from COVID-19 in the intensive care unit (ICU) (2020 to 2021) was considered. For statistical analysis, analysis of variance (ANOVA), paired samples t-test, and chi-square using SPSS-23 software were used (p<0.05). Results: Two hundred and fifty hospitalized (107 females and 143 males, mean ± standard deviation (SD) of age; 56.50 ± 17.20) patients were considered. The most (97.60%) medicine prescribed was remdesivir. Candida spp. (two females), Escherichia coli (two females), Acinetobacter spp. (one female), Citrobacter spp. (one female), Pseudomonas spp. (one male), Sphingomonas spp. (one male), Stenotrophomonas spp. (one male) and Enterobacter spp. (one male) were isolated from the patient's specimens. Four of seven bacterial isolates were positive for MDR. NI was diagnosed in six patients. There was no significant relationship between the age with the isolated microbes (P=0.154) and MDR (P=0.987) and also between gender with common microbes (P=0.576) and MDR (P=0.143). Conclusion: The coexistence of bacteria and NI was observed in patients. Remdesivir was prescribed for most patients. Most bacteria were resistant to antibiotics, especially, β-lactams. [ABSTRACT FROM AUTHOR]

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This study aimed to isolate and identify pathogenic bacteria in the intestinal tract, skin, and muscles of Sciades herzbergii; detect histopathological changes in the gill and liver; and use these biomarkers for the assessment of potential risks to human health. Fish were sampled during the rainy and dry seasons at two points in São Marcos Bay, Maranhão, Brazil: Ilha dos Caranguejos (IC) and Porto Grande (PG). Isolation and quantification were carried out using COLItest®. Colonies were subjected to identification and phenotypic investigation of antimicrobial resistance using Vitek®. Gill and liver samples were subjected to routine histological examination. The results indicated the presence of Klebsiella pneumoniae and Escherichia coli, the latter of which showed phenotypic resistance to norfloxacin and gentamicin. Fish caught at PG exhibited more extensive gill and liver damage than fish caught at IC. The findings suggest that histological changes in target organs of S. herzbergii may be influenced by infection with pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Recently, there has been a focus on the connection between the composition of gut microbiota and the development of Carbapenem-resistant Enterobacteriaceae (CRE). The objective of our work was to assess the available data on 16srRNA sequences of gut bacteria in patients admitted to the ICU. We used the ALDEx2 R package (v.1.22.0) to examine the association between these sequences and the presence and simultaneous occurrence of several bacterial species, in comparison to a control group. Out of the 60 stool samples, 24 individuals carrying CRE and 26 non-carrier groups have a higher abundance of Enterococcus durans compared to the control group. The abundance of Erysipelatoclostridium ramosum, Eubacterium limosum, Klebsiella pneumoniae, Parabacteroides distasonis, Proteus mirabilis, Bacteroides thetaiotaomicron, and Pseudomonas aeruginosa was higher in the group of individuals carrying CRE compared to non-carrier group. The CRE carrier group had a distinct profile of gut microbiota compared to the control group. These findings could contribute to advancements in the detection and treatment of antibiotic resistance patients. [ABSTRACT FROM AUTHOR]

Irritable bowel syndrome (IBS), a chronic functional gastrointestinal disorder, is recognized for its association with alterations in the gut microbiome and metabolome. This study delves into the largely unexplored domain of the gut virome in IBS patients. We conducted a comprehensive analysis of the fecal metagenomic data set from 277 IBS patients and 84 healthy controls to characterize the gut viral community. Our findings revealed a distinct gut virome in IBS patients compared to healthy individuals, marked by significant variances in between‐sample diversity and altered abundances of 127 viral operational taxonomic units (vOTUs). Specifically, 111 vOTUs, predominantly belonging to crAss‐like, Siphoviridae, Myoviridae, and Quimbyviridae families, were more abundant in IBS patients, whereas the healthy control group exhibited enrichment of 16 vOTUs from multiple families. We also investigated the interplay between the gut virome and bacteriome, identifying a correlation between IBS‐enriched bacteria like Klebsiella pneumoniae, Fusobacterium varium, and Ruminococcus gnavus, and the IBS‐associated vOTUs. Furthermore, we assessed the potential of gut viral signatures in predicting IBS, achieving a notable area under the receiver operator characteristic curve (AUC) of 0.834. These findings highlight significant shifts in the viral diversity, taxonomic distribution, and functional composition of the gut virome in IBS patients, suggesting the potential role of the gut virome in IBS pathogenesis and opening new avenues for diagnostic and therapeutic strategies targeting the gut virome in IBS management. [ABSTRACT FROM AUTHOR]

The rapid spread of carbapenemase-producing strains has led to increased levels of resistance among Gram-negative bacteria, especially enterobacteria. The current study aimed to collect and genetically characterize the colistin- and carbapenem-resistant isolates, obtained in one of the biggest hospitals (Military Medical Academy) in Sofia, Bulgaria. Clonal relatedness was detected by RAPD and MLST. Carbapenemases, ESBLs, and mgrB were investigated by PCR amplification and sequencing, replicon typing, and 16S rRNA methyltransferases with PCRs. Fourteen colistin- and carbapenem-resistant K. pneumoniae isolates were detected over five months. Six carbapenem-resistant and colistin-susceptible isolates were also included. The current work revealed a complete change in the spectrum of carbapenemases in Bulgaria. blaNDM-5 was the only NDM variant, and it was always combined with blaOXA-232. The coexistence of blaOXA-232 and blaNDM-5 was observed in 10/14 (72%) of colistin- and carbapenem-resistant K. pneumoniae isolates and three colistin-susceptible isolates. All blaNDM-5- and blaOXA-232-positive isolates belonged to the ST6260 (ST101-like) MLST type. They showed great mgrB variability and had a higher mortality rate. In addition, we observed blaOXA-232 ST14 isolates and KPC-2-producing ST101, ST16, and ST258 isolates. The colistin- and carbapenem-resistant isolates were susceptible only to cefiderocol for blaNDM-5- and blaOXA-232-positive isolates and to cefiderocol and ceftazidime/avibactam for blaOXA-232- or blaKPC-2-positive isolates. All blaOXA-232-positive isolates carried rmtB methylase and the colE replicon type. The extremely limited choice of appropriate treatment for patients infected with such isolates and their faster distribution highlight the need for urgent measures to control this situation. [ABSTRACT FROM AUTHOR]

The prevalence of multidrug-resistant Gram-negative infections, particularly carbapenem-resistant strains, has become a significant global health concern. Ceftazidime–avibactam (CZA) has emerged as a promising treatment option. However, data on its efficacy and safety in children are scarce, necessitating further investigation. We conducted a descriptive case series at a tertiary hospital in Spain from February 2019 to January 2022. Pediatric patients (<16 years) treated with CZA for confirmed or suspected multidrug-resistant Gram-negative infections were included. The clinical and microbiological characteristics, treatment approaches, and outcomes were examined. Eighteen children received CZA treatment. All had complex chronic conditions, with the most frequent underlying main diseases being liver transplantation (n = 8) and biliary atresia (n = 4). The predominant type of infection for which they received CZA was intra-abdominal infection caused or suspected to be caused by OXA-48-producing Klebsiella pneumoniae. CZA was generally well tolerated. Within the first month of starting CZA therapy, two patients died, with one case directly linked to the infection's fatal outcome. Some patients needed repeated courses of therapy due to recurrent infections, yet no resistance development was noted. In summary, the use of CZA showed effectiveness and safety, while the lack of resistance development highlights CZA's potential as a primary treatment option against OXA-48-producing infections. [ABSTRACT FROM AUTHOR]

Background: The aim of our study was to analyze the factors associated with the increased risk of urinary tract infection (UTI) with carbapenem-resistant (CR) Klebsiella pneumoniae (Kpn) and the antibiotic resistance spectrum of the strains in patients. As secondary objectives, we elaborated the profile of these patients and the incidence of different types of carbapenemases. Methods: We conducted a retrospective case-control study in which we compared a group of 62 patients with urinary tract infections with CR Kpn with a control group consisting of 136 patients with urinary tract infections with multidrug-resistant (MDR), but carbapenem-sensitive (CS), Kpn, who were hospitalized between 1 January 2022 and 31 March 2024. Results: Compared to patients with urinary tract infections with CS Kpn, patients with urinary tract infections with CR Kpn were preponderant in rural areas (62.9% vs. 47.1%, p = 0.038) and more frequently had an upper urinary tract infection (69.4% vs. 36.8%, p < 0.01). Among the risk factors examined, patients in the study group had a higher presence of urinary catheters inserted for up to one month (50% vs. 34.6%, p = 0.03), rate of hospitalization in the last 180 days (96.8% vs. 69.9%, p < 0.01) and incidence of antibiotic therapy in the last 180 days (100% vs. 64.7%, p < 0.01). They also had a higher rate of carbapenem treatment in the last 180 days (8.1% vs. 0%, p < 0.01). Patients in the study group had a broader spectrum of resistance to all antibiotics tested (p < 0.01), with the exception of sulfamethoxazole–trimethoprim, where the resistance rate was similar in both groups (80.6% vs. 67.6%, p = 0.059). In the multivariate analysis, transfer from other hospitals (OR = 3.51, 95% and CI: 1.430–8.629) and treatment with carbapenems in the last 180 days (OR = 11.779 and 95% CI: 1.274–108.952) were factors associated with an increased risk of disease compared to the control group. The presence of carbapenemases was observed in all patients with CR Kpn, in the order of frequency New Delhi metallo-ß-lactamase (NDM) (52.2%), Klebsiella pneumoniae carbapenemase (KPC) (32.6%), and carbapenem-hydrolyzing oxacillinase (Oxa-48) (15.2%). Conclusions: The environment of origin and previous treatment with carbapenems appear to be the factors associated with an increased risk of urinary tract infection with CR Kpn compared to patients with urinary tract infections with CS Kpn. CR Kpn exhibits a broad spectrum of antibiotic resistance, among which is resistance to carbapenem antibiotics. [ABSTRACT FROM AUTHOR]

Wound infection is a serious complication in burn injury, which is a common form of trauma and an important public health issue. We investigated samples from burn and nonburn wounds for microbial characteristics and temporal trends of antibiotic resistance. Wound samples were collected from 369 burned patients and 927 non-burned individuals admitted from 2007 to 2017. Higher frequency of Acinetobacter baumannii , Klebsiella pneumonia , and Pseudomonas aeruginosa was observed in samples from burned individuals when compared to those from non-burned. The prevalence of different groups of bacteria varied when the samples were stratified according to age and sex. The antimicrobial resistance profiles showed a significant difference between burned and non-burned patients. The different temporal trends of antimicrobial resistance rates were also found, which may be critical for the proper selection of antibiotics in burn treatment. The present study suggested that frequent pathogens and antibacterial resistance evolution could differ between burn wounds and other wounds. Therefore, periodic surveillance of antibiotic resistance patterns in the burn unit might help physicians properly select antibiotics for treatment. [ABSTRACT FROM AUTHOR]

Centella asiatica phytocompounds coated onto Multi‐Walled Carbon Nanotube (MWCNT) possessing antibacterial activity were prepared using a simple physical absorption method. The incorporation of Centella asiatica phytocompounds into functionalized MWCNTs could improve the physical properties of Centella asiatica phytocompounds conjugated MWCNT compared to pure MWCNT. Fourier Transform Infrared Spectroscopy (FTIR) results depict the presence of bioactive compounds on MWCNT coatings and the Raman spectrum confirms the presence of carbon in various ranges by their peaks from 636 to 1594.46 cm−1 in the MWCNT. Scanning Electron Microscope (SEM) micrograph images confirmed the existence of MWCNT‐Nano composites with Centella asiatica phytocompounds exhibiting the surface structure with pore clarity and visible pipes. Transmission Electron Microscope (TEM) images show the uniform dispersed spherical ultrathin nanosheets at the size of 10–20 nm. Further anti‐bacterial analysis results revealed that Centella asiatica phytocompounds coating enhances the anti‐bacterial activity of MWCNT. Prepared Centella asiatica phytocompounds ‐MWCNT possess maximal inhibition and produces high zones of inhibition 11 ± 0.06, 17 ± 0.08, 14 ± 0.07, and 16 ± 0.08 mm against four food‐spoiling bacterial clinical pathogens Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia respectively. Centella asiatica phytocompounds conjugated‐MWCNT spin‐coated packaging showed good preservation properties. Centella asiatica phytocompounds conjugated with MWCNTs provide antimicrobial properties, enabling the development of novel food packaging materials that can protect food from spoilage, increase food stability, and sustainability, and improve the shelf life of food. [ABSTRACT FROM AUTHOR]

Glycoconjugation is a well-established technology for vaccine development: linkage of the polysaccharide (PS) antigen to an appropriate carrier protein overcomes the limitations of PS T-independent antigens, making them effective in infants and providing immunological memory. Glycoconjugate vaccines have been successful in reducing the burden of different diseases globally. However, many pathogens still require a vaccine, and many of them display a variety of glycans on their surface that have been proposed as key antigens for the development of high-valency glycoconjugate vaccines. CDAP chemistry represents a generic conjugation strategy that is easily applied to PS with different structures. This chemistry utilizes common groups to a large range of PS and proteins, e.g., hydroxyl groups on the PS and amino groups on the protein. Here, new fast analytical tools to study CDAP reaction have been developed, and reaction conditions for PS activation and conjugation have been extensively investigated. Mathematical models have been built to identify reaction conditions to generate conjugates with wanted characteristics and successfully applied to a large number of bacterial PSs from different pathogens, e.g., Klebsiella pneumoniae, Salmonella Paratyphi A, Salmonella Enteritidis, Salmonella Typhimurium, Shighella sonnei and Shigella flexneri. Furthermore, using Salmonella Paratyphi A O-antigen and CRM197 as models, a design of experiment approach has been used to study the impact of conjugation conditions and conjugate features on immunogenicity in rabbits. The approach used can be rapidly extended to other PSs and accelerate the development of high-valency glycoconjugate vaccines. [ABSTRACT FROM AUTHOR]

Dose-intensive cytostatic therapy and antibiotic treatment in allogeneic hematopoietic stem cell transplantation (allo-HSCT) cause severe abnormalities in a composition of gut microbiota as well as the emergence of antibiotic resistance. The data on the longitudinal recovery of major bacterial phyla and the expansion of genes associated with antibiotic resistance are limited. We collected regular stool samples during the first year after allo-HSCT from 12 adult patients with oncohematological disorders after allo-HSCT and performed 16SrRNA sequencing, multiplex PCR, conventional bacteriology and CHROMagar testing. We observed a decline in Shannon microbiota diversity index as early as day 0 of allo-HSCT (p = 0.034) before any administration of antibiotics, which persisted up to 1 year after transplantation, when the Shannon index returned to pre-transplant levels (p = 0.91). The study confirmed the previously shown decline in Bacillota (Firmicutes) genera and the expansion of E. coli/Shigella, Klebsiella and Enterococci. The recovery of Firmicutes was slower than that of other phyla and occurred only a year post-transplant. A positive correlation was observed between the expansion of E. coli/Shigella genera and blaKPC, blaCTX-M-1 and blaTEM (p < 0.001), Klebsiella spp. and blaOXA-48-like, blaNDM, blaCTX-M-1, blaTEM, and blaSHV (p < 0.001), Pseudomonas spp. and blaNDM (p = 0.002), Enterococcus spp. and blaOXA-48-like, blaNDM, blaCTX-M-1, blaSHV (p < 0.01). The correlation was observed between the expansion of Enterobacterales and and carbapenemase-positive CHROMagar samples (p < 0.001). Samples positive for carbapenem-resitant bacteria were at their maximum levels on day +30, and were gradually diminishing one year after allo-HSCT. From day +30 to +60, all isolated K. pneumoniae strains in fecal samples proved to be resistant to the main antibiotic groups (carbapenems, aminoglycosides, fluoroquinolones, third-generation cephalosporins). One year after HSCT, we documented the spontaneous decolonization of K. pneumoniae. The sensitivity of molecular biology techniques in the search for total and antibiotic-resistant Klebsiella seems to be superior to common bacteriological cultures. Future studies should be focused on searching for novel approaches to the efficient reconstitution and/or maintenance of strictly anaerobic microbiota in oncological patients. [ABSTRACT FROM AUTHOR]

Introduction: In the battle against multidrug-resistant bacterial infections, ceftazidime-avibactam (CZA) stands as a pivotal defense, particularly against carbapenemresistant (CR) Gram-negative pathogens. However, the rise in resistance against this drug poses a significant threat to its effectiveness, highlighting the critical need for in-depth studies about its resistance mechanisms. Methods: This research focuses on the genomic characterization of CR- and CZA-resistant Escherichia coli (n=26) and Klebsiella pneumoniae (n=34) strains, harboring the blaNDM and/or blaOXA-48-like genes, at a major Lebanese tertiary care medical center, using whole genome sequencing (WGS). Results: Our findings revealed a notable prevalence of blaNDM in all K. pneumoniae strains isolates, with 27 of these also harboring blaOXA-48.On the other hand, E. coli strains predominantly carried the blaNDM-5 gene. Whole genome sequencing (WGS) identified a predominance of ST383 among K. pneumoniae strains, which possessed a multi-replicon IncFIB-IncHI1B plasmid harboring the blaNDM-5. Additionally, various Inc group plasmids in K. pneumoniae across multiple sequence types were found to carry the blaNDM. Similarly, diverse STs of E. coli were observed to carry blaNDM-5 on different plasmids. Discussion: The study underscores NDM carbapenemases as a paramount resistance mechanism in Lebanon, jeopardizing critical last-resort treatments. It also illuminates the role of varied sequence types and mobile genetic elements in the spread of NDM resistance, stressing the urgent need for strategies to mitigate this threat, especially in nosocomial infections. [ABSTRACT FROM AUTHOR]

The species included in the ESKAPE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and the genus Enterobacter) have a high capacity to develop antimicrobial resistance (AMR), a health problem that is already among the leading causes of death and could kill 10 million people a year by 2050. The generation of new potentially therapeutic molecules has been insufficient to combat the AMR "crisis", and the World Health Organization (WHO) has stated that it will seek to promote the development of rapid diagnostic strategies. The physicochemical properties of metallic nanoparticles (MNPs) have made it possible to design biosensors capable of identifying low concentrations of ESKAPE bacteria in the short term; other systems identify antimicrobial susceptibility, and some have been designed with dual activity in situ (bacterial detection and antimicrobial activity), which suggests that, in the near future, multifunctional biosensors could exist based on MNPs capable of quickly identifying bacterial pathogens in clinical niches might become commercially available. This review focuses on the use of MNP-based systems for the rapid and accurate identification of clinically important bacterial pathogens, exhibiting the necessity for exhaustive research to achieve these objectives. This review focuses on the use of metal nanoparticle-based systems for the rapid and accurate identification of clinically important bacterial pathogens. [ABSTRACT FROM AUTHOR]

Enterobacter hormaechei, one of the species within the Enterobacter cloacae complex, is a relevant agent of healthcare-associated infections. In addition, it has gained relevance because isolates have shown the capacity to resist several antibiotics, particularly carbapenems. However, knowledge regarding colonization and virulence mechanisms of E. hormaechei has not progressed to the same extent as other Enterobacteriaceae species as Escherichia coli or Klebsiella pneumoniae. Here, we describe the presence and role of the type 3 fimbria, a chaperone-usher assembled fimbria, which was first described in Klebsiella spp., and which has been detected in other representatives of the Enterobacteriaceae family. Eight Chilean E. cloacae isolates were examined, and among them, four E. hormaechei isolates were found to produce the type 3 fimbria. These isolates were identified as E. hormaechei subsp. hoffmannii, one of the five subspecies known. A mutant E. hormaechei subsp. hoffmannii strain lacking the mrkA gene, encoding the major structural subunit, displayed a significantly reduced adherence capacity to a plastic surface and to Caco-2 cells, compared to the wild-type strain. This phenotype of reduced adherence capacity was not observed in the mutant strains complemented with the mrkA gene under the control of an inducible promoter. Therefore, these data suggest a role of the type 3 fimbria in the adherence capacity of E. hormaechei subsp. hoffmannii. A screening in E. hormaechei genomes contained in the NCBI RefSeq Assembly database indicated that the overall presence of the type 3 fimbria is uncommon (5.94–7.37%), although genes encoding the structure were detected in representatives of the five E. hormaechei subspecies. Exploration of complete genomes indicates that, in most of the cases, the mrkABCDF locus, encoding the type 3 fimbria, is located in plasmids. Furthermore, sequence types currently found in healthcare-associated infections were found to harbor genes encoding the type 3 fimbria, mainly ST145, ST78, ST118, ST168, ST66, ST93, and ST171. Thus, although the type 3 fimbria is not widespread among the species, it might be a determinant of fitness for a subset of E. hormaechei representatives. [ABSTRACT FROM AUTHOR]

Rapid and accurate detection of Klebsiella pneumoniae carbapenem resistance is important for infection control and targeted antibiotic therapy. PCR-based assay performance heavily depends on the quality and quantity of template DNA. Challenges arise from the necessity to isolate chromosomal and large plasmid-encoded resistance genes simultaneously from a limited number of target cells and to remove PCR inhibitors. qPCRs for the detection of K. pneumoniae strains carrying blaOXA-48, blaNDM-1, blaKPC-2, and blaVIM-1 carbapenemase genes were developed. We compared the performance of template DNA extracted with silica column-based methods, reversed elution systems, and lysis-only methods either from diluted culture fluid or from a synthetic stool matrix which contained PCR inhibitors typically present in stool. The synthetic stool matrix was chosen to mimic K. pneumoniae containing rectal swabs or stool samples in a reproducible manner. For total DNA isolated from culture fluid, resistance gene detection by qPCR was always possible, independent of the extraction method. However, when total DNA was isolated from synthetic stool matrix spiked with K. pneumoniae, most methods were insufficient. The best performance of template DNA was obtained with reversed elution. This highlights the importance of choosing the right DNA extraction method for consistent carbapenem resistance detection by PCR. [ABSTRACT FROM AUTHOR]

This article discusses a scientific study conducted in 2010 by Alastruey-Izquierdo et al. The study focuses on the antifungal susceptibility profile of human-pathogenic species of Lichtheimia, a genus of fungi. The study aims to determine the minimum inhibitory concentrations of antifungal agents for these fungi. The article references a technical note by the Subcommittee on Antifungal Susceptibility Testing of the ESCMID in 2008, which provides a method for determining these concentrations. The study's findings and methodology can be explored further by accessing the full article. [Extracted from the article]

Klebsiella pneumoniae liver abscess (KPLA) is an invasive infectious disease with a considerable prevalence and complications. This study aimed to determine the predicted value of aspartate aminotransferase-to-platelet ratio index (APRI) and fibrosis-4 index (FIB-4) for positive blood cultures and sepsis in patients with KPLA. Methods: We evaluated 248 consecutive participants diagnosed with KPLA. Demographic characteristics, clinical features, and laboratory test results of the participants were recorded. Multivariate logistic regression analysis was performed to identify the risk factors. Receiver operating characteristic (ROC) analyses were conducted to evaluate the discriminatory ability of APRI and FIB-4. Results: 30.2% (75 of 248) KPLA patients presented with positive blood cultures, and 70 (28.2%) developed sepsis. Among the positive blood culture and sepsis groups, the APRI and FIB-4 showed statistically significant increases. Multivariate analysis showed that APRI (odds ratio [OR] = 1.190, 95% confidence interval [CI] 1.051– 1.347, p = 0.006) and FIB-4 (OR = 1.110, 95% CI 1.046– 1.179, p = 0.001) were independent prognostic factors for positive blood cultures. Both APRI (OR = 1.505, 95% CI 1.149-1.988, p = 0.004) and FIB-4 (OR = 1.187, 95% CI 1.054-1.336, p = 0.005) were independent risk factors for sepsis. The area under the ROC curve (AUC) of APRI and FIB-4 for predicting positive blood cultures of KPLA was 0.783 and 0.766, respectively. APRI had an AUC of 0.801, with a sensitivity and specificity of  71.4% and  81.5%, respectively, for predicting sepsis in patients with KPLA, and the prediction performance of APRI was better than that of FIB-4 (AUC = 0.798). Conclusion: In our study, APRI and FIB-4 are effective methods for predicting KPLA patients with positive blood cultures and sepsis. [ABSTRACT FROM AUTHOR]

Introduction: New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae (NDM-1-KP) sequence type (ST) 147 poses a significant threat in clinical settings due to its evolution into two distinct directions: hypervirulence and carbapenem resistance. Hypervirulence results from a range of virulence factors, while carbapenem resistance stems from complex biological mechanisms. The NDM-1-KP ST147 clone has emerged as a recent addition to the family of successful clones within the species. Methods: In this study, we successfully synthesized 5-bromo-N-alkylthiophene-2-sulfonamides (3a-c) by reacting 5-bromothiophene-2-sulfonamide (1) with various alkyl bromides (2) using LiH. We also synthesized a series of compounds (4a-g) from compound (3b) using the Suzuki-Miyaura cross-coupling reaction with fair to good yields (56– 72%). Further, we screened the synthesized molecules against clinically isolated New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. Subsequently, we conducted in-silico tests on compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I. Results: The compound (3b) with favourable drug candidate status, MIC of 0.39 μg/mL, and MBC of 0.78 μg/mL. This low molecular weight compound exhibited the highest potency against the resistant bacterial strains. The in-silico tests revealed that the compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I demonstrated H-bond and hydrophobic interactions. Conclusion: The 5-bromo-N-alkylthiophene-2-sulfonamides displayed antibacterial efficacy against New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. After the in-vivo trial, this substance might offer an alternative therapeutic option. [ABSTRACT FROM AUTHOR]