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With the rapid increase in traffic demand, the need for flexible and robust solutions for network connectivity has become crucial for transport network operators. Additionally, it is essential to control and encrypt data shared between headquarters (HQs) and local branches before transmission to the wide area network (WAN). As a result, software-defined wide area networks (SD-WANs) have gained significant attention for their ability to ensure high-quality experiences and enhance link reliability. However, despite the well-documented advantages of the SD-WAN approach, deploying these techniques in highly dynamic environments poses substantial challenges for researchers. These challenges encompass routing issues, network survivability and traffic management, largely due to fluctuations in the underlying link state, the number of routers and the constant evolution of network topology. In this context, this paper presents a comprehensive survey of SD-WAN addressing its key problems. This study takes researchers on a fascinating technological journey that provides an in-depth description of SD-WAN development, considering contributions of the industrial sector and academic projects. Firstly, the key components of an SD-WAN along with technological aspects related to the development and configuration of SD-WAN are systematically presented. Thereafter, the motivation, scope and organization of the problem space are presented along with the performance analysis of existing research on this topic. Finally, we discuss the future challenges and major issues in SD-WAN techniques. To the best of our knowledge, this paper constitutes the first attempt at a fundamental survey of SD-WAN solutions, which more adequately describes the state of the art of this specific subject. [ABSTRACT FROM AUTHOR]

Background Antimicrobial-resistant (AMR) pathogens represent an ongoing global health burden. Colonization is often a prerequisite for infection, but the risk of infection after AMR colonization is not well understood. Using population-level health administrative data, we sought to investigate the risk of infection with the same AMR organism after detection of colonization. Methods We conducted a retrospective population-wide cohort study among residents of Ontario, Canada, over a 5-year period to determine the risk of infection after detection of colonization with the following AMR pathogens: methicillin-resistant Staphylococcus aureus , vancomycin-resistant Enterococcus , extended-spectrum β-lactamase–producing Enterobacterales, and carbapenemase-producing Enterobacterales. We also examined the effects of age, sex, and health care setting of colonization detection on subsequent infection risk. Results There were 69 998 individuals with a positive AMR pathogen surveillance test result during the study period, 15.6% of which subsequently developed a sterile or nonsterile site infection within a median 57 days (IQR, 11–228). Infection rates varied among organisms: 18.3% for methicillin-resistant S aureus within a median 57 days (IQR, 10–239), 2.8% for vancomycin-resistant Enterococcus within a median 37 days (IQR, 11–119), 21.5% for extended-spectrum β-lactamase–producing Enterobacterales within a median 71 days (IQR, 18–231), and 20.3% for carbapenemase-producing Enterobacterales within a median 10 days (IQR, 3–42). A positive surveillance test result detected in a hospital was associated with increased infection risk after colonization as compared with the community setting. Conclusions The overall infection rate after colonization with an AMR pathogen was high for most organisms, highlighting the importance of detecting colonization from both an infection control and empiric antibiotic selection perspective. [ABSTRACT FROM AUTHOR]

The number of new vulnerabilities continues to rise significantly each year. Simultaneously, vulnerability databases have challenges in promptly sharing new security events with enough information to improve protections against emerging cyberattack vectors and possible exploits. In this context, several organizations adopt strategies to protect their data, technologies, and infrastructures from cyberattacks by implementing anticipatory and proactive approaches to their system security activities. To this end, vulnerability management systems play a crucial role in mitigating the impact of cyberattacks by identifying potential vulnerabilities within an organization and alerting cyber teams. However, the effectiveness of these systems, which employ multiple methods and techniques to identify weaknesses, relies heavily on the accuracy of published security events. For this reason, we introduce a discussion concerning existing vulnerability detection methods through an in-depth literature study of several research papers. Based on the results, this paper points out some issues related to vulnerability databases handling that impact the effectiveness of certain vulnerability identification methods. Furthermore, after summarizing the existing methodologies, this study classifies them into four approaches and discusses the challenges, findings, and potential research directions. [ABSTRACT FROM AUTHOR]

Abstract Background Multidrug-resistant organisms (MDRO) pose a significant threat to public health worldwide. The ability to identify antimicrobial resistance determinants, to assess changes in molecular types, and to detect transmission are essential for surveillance and infection prevention of MDRO. Molecular characterization based on long-read sequencing has emerged as a promising alternative to short-read sequencing. The aim of this study was to characterize MDRO for surveillance and transmission studies based on long-read sequencing only. Methods Genomic DNA of 356 MDRO was automatically extracted using the Maxwell-RSC48. The MDRO included 106 Klebsiella pneumoniae isolates, 85 Escherichia coli, 15 Enterobacter cloacae complex, 10 Citrobacter freundii, 34 Pseudomonas aeruginosa, 16 Acinetobacter baumannii, and 69 methicillin-resistant Staphylococcus aureus (MRSA), of which 24 were from an outbreak. MDRO were sequenced using both short-read (Illumina NextSeq 550) and long-read (Nanopore Rapid Barcoding Kit-24-V14, R10.4.1) whole-genome sequencing (WGS). Basecalling was performed for two distinct models using Dorado-0.3.2 duplex mode. Long-read data was assembled using Flye, Canu, Miniasm, Unicycler, Necat, Raven, and Redbean assemblers. Long-read WGS data with > 40 × coverage was used for multi-locus sequence typing (MLST), whole-genome MLST (wgMLST), whole-genome single-nucleotide polymorphisms (wgSNP), in silico multiple locus variable-number of tandem repeat analysis (iMLVA) for MRSA, and identification of resistance genes (ABRicate). Results Comparison of wgMLST profiles based on long-read and short-read WGS data revealed > 95% of wgMLST profiles within the species-specific cluster cut-off, except for P. aeruginosa. The wgMLST profiles obtained by long-read and short-read WGS differed only one to nine wgMLST alleles or SNPs for K. pneumoniae, E. coli, E. cloacae complex, C. freundii, A. baumannii complex, and MRSA. For P. aeruginosa, differences were up to 27 wgMLST alleles between long-read and short-read wgMLST and 0–10 SNPs. MLST sequence types and iMLVA types were concordant between long-read and short-read WGS data and conventional MLVA typing. Antimicrobial resistance genes were detected in long-read sequencing data with high sensitivity/specificity (92–100%/99–100%). Long-read sequencing enabled analysis of an MRSA outbreak. Conclusions We demonstrate that molecular characterization of automatically extracted DNA followed by long-read sequencing is as accurate compared to short-read sequencing and suitable for typing and outbreak analysis as part of genomic surveillance of MDRO. However, the analysis of P. aeruginosa requires further improvement which may be obtained by other basecalling algorithms. The low implementation costs and rapid library preparation for long-read sequencing of MDRO extends its applicability to resource-constrained settings and low-income countries worldwide.

The number of new vulnerabilities continues to rise significantly each year. Simultaneously, vulnerability databases have challenges in promptly sharing new security events with enough information to improve protections against emerging cyberattack vectors and possible exploits. In this context, several organizations adopt strategies to protect their data, technologies, and infrastructures from cyberattacks by implementing anticipatory and proactive approaches to their system security activities. To this end, vulnerability management systems play a crucial role in mitigating the impact of cyberattacks by identifying potential vulnerabilities within an organization and alerting cyber teams. However, the effectiveness of these systems, which employ multiple methods and techniques to identify weaknesses, relies heavily on the accuracy of published security events. For this reason, we introduce a discussion concerning existing vulnerability detection methods through an in-depth literature study of several research papers. Based on the results, this paper points out some issues related to vulnerability databases handling that impact the effectiveness of certain vulnerability identification methods. Furthermore, after summarizing the existing methodologies, this study classifies them into four approaches and discusses the challenges, findings, and potential research directions.

Background: Multidrug-resistant organisms (MDRO) pose a significant threat to public health worldwide. The ability to identify antimicrobial resistance determinants, to assess changes in molecular types, and to detect transmission are essential for surveillance and infection prevention of MDRO. Molecular characterization based on long-read sequencing has emerged as a promising alternative to short-read sequencing. The aim of this study was to characterize MDRO for surveillance and transmission studies based on long-read sequencing only. Methods: Genomic DNA of 356 MDRO was automatically extracted using the Maxwell-RSC48. The MDRO included 106 Klebsiella pneumoniae isolates, 85 Escherichia coli, 15 Enterobacter cloacae complex, 10 Citrobacter freundii, 34 Pseudomonas aeruginosa, 16 Acinetobacter baumannii, and 69 methicillin-resistant Staphylococcus aureus (MRSA), of which 24 were from an outbreak. MDRO were sequenced using both short-read (Illumina NextSeq 550) and long-read (Nanopore Rapid Barcoding Kit-24-V14, R10.4.1) whole-genome sequencing (WGS). Basecalling was performed for two distinct models using Dorado-0.3.2 duplex mode. Long-read data was assembled using Flye, Canu, Miniasm, Unicycler, Necat, Raven, and Redbean assemblers. Long-read WGS data with > 40 × coverage was used for multi-locus sequence typing (MLST), whole-genome MLST (wgMLST), whole-genome single-nucleotide polymorphisms (wgSNP), in silico multiple locus variable-number of tandem repeat analysis (iMLVA) for MRSA, and identification of resistance genes (ABRicate). Results: Comparison of wgMLST profiles based on long-read and short-read WGS data revealed > 95% of wgMLST profiles within the species-specific cluster cut-off, except for P. aeruginosa. The wgMLST profiles obtained by long-read and short-read WGS differed only one to nine wgMLST alleles or SNPs for K. pneumoniae, E. coli, E. cloacae complex, C. freundii, A. baumannii complex, and MRSA. For P. aeruginosa, differences were up to 27 wgMLST alleles between long-read and short-read wgMLST and 0–10 SNPs. MLST sequence types and iMLVA types were concordant between long-read and short-read WGS data and conventional MLVA typing. Antimicrobial resistance genes were detected in long-read sequencing data with high sensitivity/specificity (92–100%/99–100%). Long-read sequencing enabled analysis of an MRSA outbreak. Conclusions: We demonstrate that molecular characterization of automatically extracted DNA followed by long-read sequencing is as accurate compared to short-read sequencing and suitable for typing and outbreak analysis as part of genomic surveillance of MDRO. However, the analysis of P. aeruginosa requires further improvement which may be obtained by other basecalling algorithms. The low implementation costs and rapid library preparation for long-read sequencing of MDRO extends its applicability to resource-constrained settings and low-income countries worldwide. [ABSTRACT FROM AUTHOR]

This study reports a wave of abortions, diarrhea and a sharp drop in milk production at a dairy farm. The farm was routinely vaccinated with an imported vaccine against BVDV, containing the typical strains of Pestiviruses (BVDV-1 and BVDV-2). A total of 13 serum samples, 5 vaginal discharge samples, and 5 fecal samples were collected from aborted cows and from cows showing persistent diarrhea. All the samples were screened for potential microbial causes of abortion (viral or bacterial) using PCR. Out of 23 samples tested, only one vaginal discharge sample yielded a positive PCR result at the expected size of 288 bp. The primers used were designed with high sensitivity to 5'-UTR-based-RTPCR assays for the detection of Pestiviruses. The PCR product was sent for sequence analysis, and results were submitted to GenBank Accession Number #OR425033, and designed as GERD/VSVRI/pesti- Giraffe/2022. The virus was then successfully isolated and propagated in MDBK cells through three successive blind passages. A distinct cytopathic effect (CPE) was observed 2- 3 days after an incubation period post-virus inoculation, characterized by vacuolation, cell rounding, and cluster formation 72 hours after infection. PCR was carried out on each passage and gave a specific band at the expected size. Further analysis through sequence alignment and phylogenetic analysis revealed that the isolate is closely related to the Pestivirus giraffe, especially Pestivirus PG-2. This marks the first record of detection, isolation, and characterization of this strain in Egypt. Therefore, this epidemic was caused by a newly introduced strain recorded in Egypt. So, the imported vaccines would not provide protection, and the local vaccine need to be updated to include this Pestivirus strains. [ABSTRACT FROM AUTHOR]

This article discusses the importance of complete panicle exsertion (CPE) in rice, which refers to the process of the panicle fully emerging from the leaf sheath. The study utilized induced mutagenesis and molecular breeding strategies to uncover the genetic regulation of CPE in rice. The researchers identified novel genomic loci and gene alleles that have significant implications for improving rice crop yield and food security. The findings suggest that these genetic variations can be used in marker-assisted breeding and genome editing to develop high-yielding rice varieties with the desired CPE trait. [Extracted from the article]

Infections by multidrug-resistant organisms (MDROs) pose significant public health challenges, including increased mortality rates, healthcare costs, and significant impacts on the quality of life for patients. Utilizing a systematic review methodology adhering to PRISMA guidelines, we performed a comprehensive search across three databases, identifying 20 relevant studies that investigated the psychological effects of infections due to MDROs on hospitalized adults. The primary outcomes examined included depression, anxiety, and other psychosocial impacts, while secondary outcomes included patient and caregiver understanding of the infection. Findings revealed consistent associations between contact isolation due to MDRO infections and heightened levels of depression and anxiety among patients, although evidence regarding the impact on anger was mixed. Other psychological aspects, such as feelings of stigmatization and reduced healthcare provider interactions, were also recorded. The current systematic review highlights the importance of addressing these psychological effects through holistic, patient-centered care approaches, emphasizing the need for better communication and comprehensive education for both patients and healthcare providers. Our findings suggest that mitigating the psychological burden of MDROs can enhance overall patient care and outcomes and call for further research to optimize care strategies for patients hospitalized for infections due to MDROs. [ABSTRACT FROM AUTHOR]

The electrical percolation of alternating current through two-phase polyethylene/graphite composite electrodes with different contents of graphite microparticles immersed in aqueous KCl solutions has been studied. Above the graphite content of the first percolation threshold, the electrochemical impedance response of this electrode is associated with an equivalent circuit of resistance R u in series with a constant phase element (CPE). An insulator material + conducting filler model is proposed in which the electroactive surface is considered as the intersection of the percolation cluster through the solid and the cluster associated with the interfacial region. CPE is analyzed assuming a distribution of microcapacitors of the graphite particles in contact with the dielectric solution and inside the dielectric polymeric phase. [ABSTRACT FROM AUTHOR]

ABSTRACT Carbapenemase-producing Enterobacterales (CPE) isolates increasingly carry oxacillinase-48 (OXA-48)-like enzymes encoded by blaOXA-48-like, which can confer high levels of carbapenem resistance. This aims to determine the prevalence of CPE and genetic variation among blaOXA-48-like-carrying isolates recovered from Prapokklao Hospital in Chanthaburi Province, Thailand in 2016–2017. In total, 122 carbapenem-resistant Enterobacterales (CRE) isolates were recovered from clinical samples. CRE were evaluated using standard biochemical tests and MIC test strips. Carbapenemase production was assessed through the modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), and EDTA-modified carbapenem inactivation method (eCIM). Detection of blaOXA-48-like mutations was conducted via PCR and confirmed by Sanger sequencing. Among these CRE isolates, 72 (59.02%), 44 (36.07%), 3 (2.46%), and 3 (2.46%) were Klebsiella pneumoniae, Escherichia coli, Enterobacter aerogenes, and Enterobacter cloacae, respectively. The MHT identified carbapenemase production in 108 isolates (88.52%). Based on the mCIM, 81 isolates (66.39%) were carbapenemase producers. Seventy-three isolates (59.84%) were eCIM-positive, indicating metallo-β-lactamase production. Three distinct genetic variants of the blaOXA-48-like gene were identified among the isolates, including the wild-type and two point mutation types harboring the mutations E168Q and S171A (mutation type 1) and E168Q, S171A, and R214S (mutation type 2). Multiple-sequence alignment and in silico analysis revealed variation of R214 located in the β5–β6 loop. This study identified blaOXA-48-like point mutation groups and carbapenemase production, predominantly metallo-β-lactamases, among CRE isolates, especially K. pneumoniae and E. coli. These findings highlight the importance of implementing stringent infection control measures and active antimicrobial resistance surveillance to combat the spread of difficult-to-treat, metallo-β-lactamase-producing CRE in healthcare settings.IMPORTANCEIn this study, we aimed to investigate genetic variation and CPE among blaOXA-48-like carrying isolates recovered from Prapokklao Hospital, Chanthaburi Province, Thailand, during 2016–2017. A total of 122 carbapenem-resistant Enterobacterales (CRE) were recovered from clinical samples in Prapokklao Hospital. All CRE samples were confirmed by standard biochemical tests and minimum inhibitory concentration (MIC) test strips (E-test). The carbapenemase production was determined using the modified Hodge test (MHT), the modified carbapenem inactivation method (mCIM), and EDTA-CIM (eCIM). Three single mutations (E168Q, S171A, and R214S) were characterized in this study. This mutation might reflect the hydrolysis of the modified β-lactam spectrum, especially carbapenem, by OXA-48-like. Our report provides evidence of the blaOXA-48-like point mutation and carbapenemase-producing phenotype of CRE detected in this healthcare setting. Effective control measures and active surveillance of drug resistance in nosocomial pathogens are crucial for controlling diseases associated with difficult-to-treat bacteria.

Seasonal influenza A virus subtype H3N2 (A/H3N2) circulates globally and has been linked to higher hospitalization rates and summer outbreaks in temperate regions. Here, A/H3N2 circulation in Shanghai, China was systematically studied using data and materials generated by the Shanghai influenza surveillance network from 2005 to 2023. Time-series analysis of incidence and subtyping data showed that A/H3N2 co-circulated with other (sub)types and dominated in multiple seasonal influenza peaks, preferentially in summer. Whole genomes of 528 representative strains were sequenced, and spatiotemporal phylodynamic analysis using these and GISAID-archived sequences demonstrated that in the years before the COVID-19 pandemic, phylogenetically similar strains were circulating locally and elsewhere. However, clade 1a.1 (within 3C.2a.1b.2a), circulated in and only in Shanghai and domestically in 2022, while the sibling clade 2 predominated in other regions. Interestingly, clade 1a.1 was swiftly and completely replaced by clade 2, mostly 2a.3a.1, at the start of 2023. In hemagglutination inhibition and neutralization assays, sera from healthy donors collected in 2022 displayed higher or similar reactivity against 2a.3a.1 compared to 1a.1. By contrast, transcription and replication competence of 2a.3a.1 in MDCK cells was higher than 1a.1. These results indicated that instead of antigenicity differences enabling evasion of pre-existing immunity, higher replicative capability more likely contributed to 2a.3a.1 viruses achieving dominance in China. In addition to summarizing patterns of A/H3N2 local circulation in Shanghai, this work revealed an unusual episode in A/H3N2 global circulation and evolution dynamics in connection to the COVID-19 pandemic and explored possible mechanistic explanations.

Abstract Electrification mode is one of the important factors affecting the energy consumption and drainage effect of the electroosmosis method. In this work, a series of laboratory tests were performed to explore the impact of continuous changes in the electric field on the migration of ions and electrons, as well as the migration of electroosmotic flow. The results show that different electrification modes had a significant impact on the current and water discharge. An electrode reversed from anode to cathode, from one circuit to another, is beneficial to drainage, while an electrode reversed from cathode to anode, from one circuit to another, is unfavorable to drainage. The drainage effect of the specific cyclic and progressive electroosmosis (CPE) is not as good as the conventional electroosmosis method. However, combining the two electrification modes can further enhance the shear strength of the soil near a designated electrode. The generation of transverse cracks in soil is mainly due to the drainage difference caused by the electroosmotic flow, which is caused by the adjacent electric field at that location. When the electroosmotic flow is strong, longitudinal cracks will occur in the soil along the strongest electroosmotic flow path. In practical in situ engineering applications, the electrification mode of CPE will be more energy‐efficient than conventional electrification modes with the same processing time.

The timing of the grape harvest is a critical decision for winemakers, as it greatly impacts the quality and organoleptic characteristics of the resulting red wines. One key indicator for determining the optimal harvest time is the phenolic content of the grapes. As the berries ripen, phenolic compounds in the grape skin cells migrate from the seeds to the pulp and finally to the skin. However, monitoring these phenolic changes, particularly in the seeds, presents a challenge. This research addresses this problem by developing a novel technique to track the phenolic composition of seeds during ripening. The objective is to provide a reliable method for winemakers to monitor the phenolic evolution and improve harvest decision-making.In this study, a multisensory system consisting of four electrochemical enzymatic carbon paste sensors, modified with tyrosinase and electrocatalytic materials (AuNPs, lutetium phthalocyanine, and nickel oxide nanoparticles), was employed to analyze the phenolic content in grape seed extracts. The system monitored weekly changes in the phenolic composition of the seeds from three red grape varieties—Cabernet Sauvignon, Tempranillo, and Prieto Picudo—during their ripening from veraison to being overripe. Using voltammetric techniques, the electrochemical responses were characterized by shifts in peak positions and intensity changes, reflecting the oxidation/reduction of phenols. Principal Component Analysis (PCA) demonstrated the ability of the array of sensors to discriminate phenolic changes across ripening stages, while Partial Least Squares (PLS) regression provided robust correlation models between the electrochemical responses and seed phenolic content, with correlation coefficients ranging from 0.93 to 0.99. The developed methodology successfully tracked phenolic changes, offering a promising tool for monitoring grape seed maturation and assisting in determining the optimal harvest time.

This work investigates the AC conductivity, impedance spectroscopy, and modulus formulation of varied compositions of glass samples within the modified borovanadate glassy system, synthesized using the melt-quenching technique. Up to 10 mol% of BaO, the AC conductivity shows a rising trend as BaO content rises; however, for x = 15&20 mol%, there is a drop. The Almond West model fits the experimental data of ac conductivity and parameters such as crossover frequency, frequency exponent (s), and direct current conductivity are retrieved. All samples feature a CBH conduction mechanism at varying frequencies, except for VBa1, which has a small polaron quantum mechanical mechanism. There is good agreement in the activation energy derived from conductivity (0.565–0.742 eV), electric modulus (0.526–0.616 eV), and impedance research (0.530–0.743 eV). Dielectric studies reveal non-Debye-type behavior. According to impedance formalism, electric conduction occurs through Ba2+ ions and electron/polarons. This characteristic renders the glasses well-suited for energy storage devices. [ABSTRACT FROM AUTHOR]

The prevalence of Clostridium perfringens spores in the environment is crucial for the pathogenesis of this bacterium because these dormant spores, upon contact with a suitable host, can return to active growth to cause disease. We evaluated the prevalence of C. perfringens spores in the Hail and Qassim regions of Saudi Arabia. Methods: A plating method was used to identify C. perfringens. PCR analyses and DNA sequencing were performed for genotypic characterization of the newly isolated C. perfringens.25 of 300 collected samples were identified as C. perfringens by selecting black colonies on selective media and monitoring b-hemolysis on blood agar plate. PCR analyses showed that all 25 isolates carry a-toxin gene (plc), but not the enterotoxin gene (cpe), further confirming that these isolates are indeed cpe-negative. Genome sequencing found that Saudi isolates are genotypically similar to the American and European isolates; no homologues of toxinotyping genes encoding b-, i-, e- and NetB-toxin were detected in 22 Saudi isolates, with the exception of 3 isolates that carry only e-toxin gene etx. Collectively, our findings suggest that C. perfringens are highly prevalent in the Hail and Qassim environment, with 22 (88%) of 25 isolates are type A and remaining 3 (12%) are type D. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this systematic review was to investigate the effect of antibiotics on the eradication of multidrug-resistant organisms (MRO) in intestinal carriers. We defined multidrug-resistant organisms as vancomycin-resistant Enterococcus faecium (VREfm), and multidrug-resistant Gram-negative Enterobacterales. Methods: We searched the EMBASE, Cochrane Central, and PubMed databases from inception to medio November 2023. We included randomised and controlled clinical trials (RCTs), that investigated the effect of antibiotics on the eradication of multidrug-resistant organisms in intestinal carriers. Finally, we performed a meta-analysis. Results: We included five RTCs in the systematic review. In four studies an effect of antibiotics on the eradication of MRO was shown at the end of intervention, but it was not sustained at follow-up. In the fifth study, the effect at the end of intervention was not reported, and there was no observed effect of the intervention at follow-up. We included four studies in the meta-analysis, and it suggests an effect of antibiotics on the eradication of MRO in intestinal carriers at the end of follow-up with a p-value of 0.04 (95% confidence interval 1.02–1.95). None of the studies reported a significant increase in resistance to the study drug. Gastrointestinal disorders were the most frequent non-severe adverse event. Conclusions: The effect of antibiotics on the eradication of multidrug-resistant organisms in intestinal carriers was not statistically significant in any of the five included studies; however, we found a significant effect in the pooled meta-analysis. As the confidence interval is large, we cannot determine the clinical importance of this finding, and it should be further investigated. [ABSTRACT FROM AUTHOR]

Although the strain rate sensitivity of polymers processed by traditional technologies like injection molding has been extensively studied in the literature, limited research studies exist examining this sensitivity for fused deposition modeling (FDM). As such, in an exploratory approach, this study illuminates the strain rate sensitivity of five thermoplastic materials under various tensile test speeds, from 2.5 to 400 mm/min. This study aims to analyze strain rate influence on mechanical characteristics of FDM 3D-printed materials, particularly CPE+ and nylon. PLA, ABS, and PC are also used as additional materials for comparison purposes. The influence of strain rate on tensile strength, yield strength, toughness, and fracture morphology is examined. The outcome of this investigation can be used to better understand how FDM parts, made of various thermoplastic materials, behave when dynamic loading is present. [ABSTRACT FROM AUTHOR]

Electrification mode is one of the important factors affecting the energy consumption and drainage effect of the electroosmosis method. In this work, a series of laboratory tests were performed to explore the impact of continuous changes in the electric field on the migration of ions and electrons, as well as the migration of electroosmotic flow. The results show that different electrification modes had a significant impact on the current and water discharge. An electrode reversed from anode to cathode, from one circuit to another, is beneficial to drainage, while an electrode reversed from cathode to anode, from one circuit to another, is unfavorable to drainage. The drainage effect of the specific cyclic and progressive electroosmosis (CPE) is not as good as the conventional electroosmosis method. However, combining the two electrification modes can further enhance the shear strength of the soil near a designated electrode. The generation of transverse cracks in soil is mainly due to the drainage difference caused by the electroosmotic flow, which is caused by the adjacent electric field at that location. When the electroosmotic flow is strong, longitudinal cracks will occur in the soil along the strongest electroosmotic flow path. In practical in situ engineering applications, the electrification mode of CPE will be more energy‐efficient than conventional electrification modes with the same processing time. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Enterobacterales (CRE) causing severe infections in humans have represented an important challenge for clinicians worldwide during the past two decades. Novel β-lactams and β-lactam/β-lactamase inhibitor combinations have led to a shift in the first-line approach to the treatment of severe CRE infections from polymyxin-based regimens to treatment with less toxic agents. This new scenario offers the opportunity to apply rapid molecular diagnostic tests for CRE infection to identify different types of carbapenemases. Herein, the authors provide an overview of this subject and follow it with their expert perspectives. When considering studies actually measuring the clinical impact of rapid molecular tests in real-life scenarios, high certainty evidence from randomized controlled trials is still limited and not focused on CRE infections. Nonetheless, it is indisputable that rapid molecular tests have been shown to impact early therapeutic choices (in terms of both escalation and de-escalation) when used in real-life settings, thus issues in the clinical interpretation of their results are already relevant. Overall, increased expertise is required for the appropriate interpretation of rapid molecular tests for personalized antibiotic selection by understanding their strengths and limitations. [ABSTRACT FROM AUTHOR]

Ageing is an inevitable biological process characterized by progressive decline in physiological functions. It is a complex natural phenomenon that will cause structural and functional decline. Despite substantial progress in understanding the mechanism of ageing, both predictive biomarkers and preventive therapies remain limited. Using Weighted Gene Co‐expression Network Analysis (WGCNA) and machine learning techniques, we identified Carboxypeptidase E (CPE) as a pivotal marker of skin ageing, based on ageing‐related bulk transcriptome and single‐cell transcriptome data. Next, our investigation reveals downregulation of CPE in replicative, UVA‐induced, and H2O2‐induced senescent human dermal fibroblast cells (HDFs). Furthermore, shRNA‐mediated CPE knockdown induced HDFs senescence, and overexpression of CPE delayed HDFs senescence. Moreover, downregulated CPE inhibits collagen synthesis and induces inflammation, highlighting its potential as a therapeutic target for skin ageing. In conclusion, our study demonstrated that CPE functions as a predictor and optional target for therapeutic intervention of skin ageing. [ABSTRACT FROM AUTHOR]

Clostridium perfringens enterotoxin (Cpe)-producing strains cause gastrointestinal infections in humans and account for the second-largest number of all foodborne outbreaks caused by bacterial toxins. The Cpe toxin is only produced during sporulation; this process might be affected when C. perfringens comes into contact with host cells. The current study determined how the cpe expression levels and spore formation changed over time during co-culture with Caco-2 cells (as a model of intestinal epithelial cells). In co-culture with Caco-2 cells, total C. perfringens cell counts first decreased and then remained more or less stable, whereas spore counts were stable over the whole incubation period. The cpe mRNA level in the co-culture with Caco-2 cells increased more rapidly than in the absence of Caco-2 cells (3.9-fold higher levels in coculture than in the absence of Caco-2 cells after 8 h of incubation). Finally, we found that cpe expression is inhibited by a cue released by Caco-2 cells (8.3-fold lower levels in the presence of supernatants of Caco-2 cells than in the absence of the supernatants after 10 h of incubation); as a consequence, the increased expression in co-culture with Caco-2 cells must be caused by a factor associated with the Caco-2 cells. [ABSTRACT FROM AUTHOR]

Background: Carbapenemase-producing Enterobacterales (CPE) is an increasing problem in healthcare settings. This study aimed to identify the source of a CPE outbreak that occurred in 2022, in a tertiary hospital in the North of Portugal, to identify exposed patients, and to assess the risk of becoming CPE-positive following hospital admission. Methods: A multi-disciplinary investigation was conducted including descriptive, analytical, and molecular epidemiology, environmental screening, and assessment of infection control measures. Clinical and environmental isolates were analyzed using whole-genome sequencing and phylogenetic analysis. Additionally, a prospective observational cohort study was conducted to further investigate the risk factors associated with the emergence of new cases in cohorts of CPE-negative admitted patients. Results: We observed the presence of multispecies KPC-, IMP-, and/or NDM-producing isolates. Genetically indistinguishable clinical and environmental isolates were found on the same room/ward. The ST45 KPC-3-producing Klebsiella pneumoniae clone was the responsible for the outbreak. During patients’ treatment, we detected the emergence of resistance to ceftazidime-avibactam, associated with mutations in the blaKPC-3 gene (blaKPC-46, blaKPC-66 and blaKPC-124, the last variant never previously reported), suggesting a vertical evolutionary trajectory. Patients aged ≥ 75 years, hygiene/feeding-care dependent, and/or subjected to secretion aspiration were risk factors for CPE colonization after hospital admission. Additionally, cases with previous admission to the emergency department suggest that CPE dissemination may occur not only during hospitalization but also in the emergency department. Conclusion: Overall, the study highlights that selection pressure with antibiotics, like ceftazidime-avibactam, is a contributing factor to the emergence of new β-lactamase variants and antibiotic resistance. It also shows that the hospital environment can be a significant source of CPE transmission, and that routine use of infection control measures and real-time molecular epidemiology investigations are essential to ensure the long-term termination of CPE outbreaks and prevent future resurgences.

In large-scale all-solid-state storage technologies, solid polymer electrolytes (SPEs) provide greater safety and longer cycle life than traditional liquid or gel polymer electrolytes. Polymer electrolytes (PEs) derived from biopolymers have been intensively explored for use in electrochemical devices due to their great flexibility, low cost, and environmental sustainability. However, biopolymer-based electrolytes cannot meet the expectations of practical applications at room temperature due to their low ionic conductivity. Over the years, improving the performance of this class of electrolytes has been the focus of intense research and development, notably polymer blending, plasticization, and structural functionalization. Here, we investigate the performance of an all-biopolymer solid electrolytes based on a methylcellulose-pectin blend doped with potassium phosphate. FESEM micrographs, as well as the shifting and changing intensity of FTIR bands in the electrolyte specimens confirm the polyblend homogeneity with no phase separation. The increased amorphous fraction of the composite polymer electrolytes (CPEs) is seen in the XRD and DSC patterns of the plasticized and unplasticized samples. Impendence studies performed on the system recorded a maximum ionic conductivity of 1 × 10–5 Scm−1 by doping with 50 wt.% K3PO4. This value further increased to 5.9 × 10–4 Scm−1 upon adding 25 wt.% EC to the polymer system. This sample also possesses an electrochemical stability window of 4.24 V and an ion transference number of 0.95. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) is a worldwide healthcare problem. Multidrug-resistant organisms (MDROs) can spread quickly owing to their resistance mechanisms. Although colonized individuals are crucial for MDRO dissemination, colonizing microbes can lead to symptomatic infections in carriers. Carbapenemase-producing Enterobacterales (CPE) are among the most important MDROs involved in colonizations and infections with severe outcomes. This review aimed to track down the first reports of CPE in Africa, describe their dissemination throughout African countries and summarize the current status of CRE and CPE data, highlighting current knowledge and limitations of reported data. Two database queries were undertaken using Medical Subject Headings (MeSH), employing relevant keywords to identify articles that had as their topics beta-lactamases, carbapenemases and carbapenem resistance pertaining to Africa or African regions and countries. The first information on CPE could be traced back to the mid-2000s, but data for many African countries were established after 2015–2018. Information is presented chronologically for each country. Although no clear conclusions could be drawn for some countries, it was observed that CPE infections and colonizations are present in most African countries and that carbapenem-resistance levels are rising. The most common CPE involved are Klebsiella pneumoniae and Escherichia coli, and the most prevalent carbapenemases are NDM-type and OXA-48-type enzymes. Prophylactic measures, such as screening, are required to combat this phenomenon. [ABSTRACT FROM AUTHOR]

Carbapenemase-producing Enterobacterales (CPE) is an increasing problem in healthcare settings. This study aimed to identify the source of a CPE outbreak that occurred in 2022, in a tertiary hospital in the North of Portugal, to identify exposed patients, and to assess the risk of becoming CPE-positive following hospital admission. A multi-disciplinary investigation was conducted including descriptive, analytical, and molecular epidemiology, environmental screening, and assessment of infection control measures. Clinical and environmental isolates were analyzed using whole-genome sequencing and phylogenetic analysis. Additionally, a prospective observational cohort study was conducted to further investigate the risk factors associated with the emergence of new cases in cohorts of CPE-negative admitted patients. We observed the presence of multispecies KPC-, IMP-, and/or NDM-producing isolates. Genetically indistinguishable clinical and environmental isolates were found on the same room/ward. The ST45 KPC-3-producing Klebsiella pneumoniae clone was the responsible for the outbreak. During patients' treatment, we detected the emergence of resistance to ceftazidime-avibactam, associated with mutations in the bla KPC-3 gene (bla KPC-46 , bla KPC-66 and bla KPC-124 , the last variant never previously reported), suggesting a vertical evolutionary trajectory. Patients aged ≥ 75 years, hygiene/feeding-care dependent, and/or subjected to secretion aspiration were risk factors for CPE colonization after hospital admission. Additionally, cases with previous admission to the emergency department suggest that CPE dissemination may occur not only during hospitalization but also in the emergency department. Overall, the study highlights that selection pressure with antibiotics, like ceftazidime-avibactam, is a contributing factor to the emergence of new β-lactamase variants and antibiotic resistance. It also shows that the hospital environment can be a significant source of CPE transmission, and that routine use of infection control measures and real-time molecular epidemiology investigations are essential to ensure the long-term termination of CPE outbreaks and prevent future resurgences. [ABSTRACT FROM AUTHOR]

Infections by multidrug-resistant organisms (MDROs) pose significant public health challenges, including increased mortality rates, healthcare costs, and significant impacts on the quality of life for patients. Utilizing a systematic review methodology adhering to PRISMA guidelines, we performed a comprehensive search across three databases, identifying 20 relevant studies that investigated the psychological effects of infections due to MDROs on hospitalized adults. The primary outcomes examined included depression, anxiety, and other psychosocial impacts, while secondary outcomes included patient and caregiver understanding of the infection. Findings revealed consistent associations between contact isolation due to MDRO infections and heightened levels of depression and anxiety among patients, although evidence regarding the impact on anger was mixed. Other psychological aspects, such as feelings of stigmatization and reduced healthcare provider interactions, were also recorded. The current systematic review highlights the importance of addressing these psychological effects through holistic, patient-centered care approaches, emphasizing the need for better communication and comprehensive education for both patients and healthcare providers. Our findings suggest that mitigating the psychological burden of MDROs can enhance overall patient care and outcomes and call for further research to optimize care strategies for patients hospitalized for infections due to MDROs.

The rapid increase of OXA-244-producing Escherichia coli, predominantly driven by genetically clustered isolates of sequence type (ST)38, has been observed in at least nine European countries, including Germany. However, the reasons for the spread of OXA-244-producing E. coli remain unclear. Here, we aim to evaluate the possibility of prolonged carriage. We identified a total of six different patients with repeated detection of OXA-244-producing E. coli isolates, which were subjected to both short and long-read whole-genome sequencing (WGS). Besides allelic differences using core genome multilocus sequence typing (cgMLST) analyses, we obtained numbers of single-nucleotide polymorphisms (SNPs) to calculate individual base-pair substitution (BPS) rates. To assess possible re-exposure and risk factors for prolonged carriage, case interviews were conducted. The time between detections ranged from eleven months to more than three years. Initial isolates originated in three+ out of six cases from clinical samples, whereas remaining samples were from screening, mostly in the inpatient setting. As expected, cgMLST analyses showed low numbers of allelic differences between isolates of each case ranging from 1 to 4, whereas numbers of SNPs were between 2 and 99 (mean = 36), thus clearly highlighting the discrepancy between these different bacterial typing approaches. For five out of six cases, observed BPS rates suggest that patients can be colonized with OXA-244-producing E. coli, including ST38 cluster isolates, for extensively long times. Thus, we may have previously missed the epidemiological link between cases because exposure to OXA-244-producing E. coli could have occurred in a time frame, which has not been evaluated in previous investigations. Our results may help to guide future epidemiological investigations as well as to support the interpretation of genetic diversity of OXA-244-producing E. coli, particularly among ST38 cluster isolates.

Simple Summary: Diarrheal diseases in calves cause economic losses in countries worldwide. In particular, bovine coronavirus, the main cause of diarrhea in calves, results in high economic losses for cow farmers. In South Korea, calf diarrhea, which affects newborn calves, and winter dysentery, which occurs in adult cows during the winter, are detected continuously. The vaccine strain (BC94) used in South Korea belongs to the GI type; however, a phylogenetic analysis revealed that all of the prevalent circulating strains belong to the GIIa type. Therefore, we attempted to develop a live attenuated BCoV vaccine candidate that targets recent prevalent strains. The bovine coronavirus (BCoV) KBR-1 strain, obtained from calf diarrhea samples collected in 2017, belongs to group GIIa. To attenuate this strain, it was subcultured continuously (up to 79 times) in HRT-18 cells, followed by 80–120 passages in MDBK cells. The KBR-1-p120 strain harvested from MDBK cells at passage 120 harbored 13 amino acid mutations in the spike gene. Additionally, the KBR-1-p120 strain showed a high viral titer and cytopathogenic effects in MDBK cells. Seven-day-old calves (negative for BCoV antigen and antibodies) that did not consume colostrum were orally inoculated with the attenuated candidate strain (KBR-1-p120), or with KBR-1 passaged 10 times (KBR-1-p10) in HRT-18 cells. Calves inoculated with KBR-1-p10 had a low diarrhea score, and BCoV RNA was detected at 3–7 days post-inoculation (DPI). The virus was also present in the duodenum, jejunum, and ileum at autopsy; however, calves inoculated with KBR-1-p120 had low levels of BCoV RNA in feces at 4–6 DPI, and no diarrhea. In addition, an extremely small amount of BCoV RNA was present in the jejunum and ileum at autopsy. The small intestines of calves inoculated with KBR-1-p120 were emulsified and used to infect calves two more times, but pathogenicity was not recovered. Therefore, the KBR-1-p120 strain has potential as a live vaccine candidate. [ABSTRACT FROM AUTHOR]

The development of antibiotic resistance mechanisms hinders the treatment process. So far, there is limited data on the problem of bacterial resistance in hospitals in Central and Eastern Europe. Therefore, this study aimed to assess the prevalence of resistance mechanisms and alert pathogens based on reports regarding cultures of samples collected from general hospital patients in Poland in the period 2019–2021. This study examined the prevalence of resistance mechanisms and alert pathogens and the structure of microorganisms, including the type of diagnostic material in the hospital department. The frequency of occurrence and the trends were analysed based on Cochran's Q-test, relative change and the average annual rate of change (AARC). Of all 14,471 cultures, 3875 were positive for bacteria, and 737 were characterised by resistance mechanisms (19.0%). Alert pathogens were identified in 983 cases (24.6%), including pathogens isolated from blood samples. The most commonlyisolated bacteria were Escherichia coli (>20% of positive cultures), Enterococcus faecalis (7%), and Klebsiella pneumoniae (6%) increasing over time; Staphylococcus aureus (13%) was also found, but its proportion was decreasing over time. Extended-spectrum β-lactamase (ESBL) was the most frequent resistance mechanism in Internal Medicine (IM) (p < 0.001) and the Intensive Care Unit (ICU) (p < 0.01), as well as in ICU-COVID; this increased over the study period (AARC ↑34.9%). Methicillin-resistant Staphylococcus aureus (MRSA) (AARC ↓50.82%) and AmpC beta-lactamase (AARC ↓24.77%) prevalence fell over time. Also, the number of alert pathogens was dominant in the IM (p < 0.01) and ICU (p < 0.001). The most common alert pathogen was ESBL-positive E. coli. Gram-negative rods constitute a significant epidemiological problem in hospitals, especially the growing trend of ESBL in IM and ICU, which calls for increased control of sanitary procedures. [ABSTRACT FROM AUTHOR]

Altered prohormone processing, such as with proinsulin and pro-islet amyloid polypeptide (proIAPP), has been reported as an important feature of prediabetes and diabetes. Proinsulin processing includes removal of several C-terminal basic amino acids and is performed principally by the exopeptidase carboxypeptidase E (CPE), and mutations in CPE or other prohormone convertase enzymes (PC1/3 and PC2) result in hyperproinsulinemia. A comprehensive characterization of the forms and quantities of improperly processed insulin and other hormone products following Cpe deletion in pancreatic islets has yet to be attempted. In the present study we applied top-down proteomics to globally evaluate the numerous proteoforms of hormone processing intermediates in a β-cell-specific Cpe knockout mouse model. Increases in dibasic residue–containing proinsulin and other novel proteoforms of improperly processed proinsulin were found, and we could classify several processed proteoforms as novel substrates of CPE. Interestingly, some other known substrates of CPE remained unaffected despite its deletion, implying that paralogous processing enzymes such as carboxypeptidase D (CPD) can compensate for CPE loss and maintain near normal levels of hormone processing. In summary, our quantitative results from top-down proteomics of islets provide unique insights into the complexity of hormone processing products and the regulatory mechanisms. [ABSTRACT FROM AUTHOR]

This study describes a one-step, facile, simple, and effective electrochemical method for the codeposition of graphene oxide (GO) and Pd nanoparticles (NPs) onto a carbon paste electrode (CPE). The obtained nanocomposite was characterized using scanning electron microscopy, cyclic voltammetry, and chronocoulometry techniques, which confirmed the high dispersion and stability of PdNPs supported on the graphene. This procedure does not require the use of a reducing agent or surfactant, making the preparation process very clean. This catalyst showed superior electrocatalytic activity and stability toward hydrogen evolution reaction (HER) when compared to either PdNPs or graphene alone, indicating the synergistic effect of graphene and PdNPs. The Tafel slopes of HER on both bare and modified CPE were determined to be 97 and 146 mV dec-1, respectively. In addition, the kinetic parameters showed that the Volmer step must control the HER. This study suggests an effective and controllable method for preparing graphene–metal NPs with high electrocatalytic activity.

Abstract Background Accumulating evidence shows a role of the hospital wastewater system in the spread of multidrug-resistant organisms, such as carbapenemase producing Enterobacterales (CPE). Several sequential outbreaks of CPE on the geriatric ward of the Ghent University hospital have led to an outbreak investigation. Focusing on OXA-48 producing Citrobacter freundii, the most prevalent species, we aimed to track clonal relatedness using whole genome sequencing (WGS). By exploring transmission routes we wanted to improve understanding and (re)introduce targeted preventive measures. Methods Environmental screening (toilet water, sink and shower drains) was performed between 2017 and 2021. A retrospective selection was made of 53 Citrobacter freundii screening isolates (30 patients and 23 environmental samples). DNA from frozen bacterial isolates was extracted and prepped for shotgun WGS. Core genome multilocus sequence typing was performed with an in-house developed scheme using 3,004 loci. Results The CPE positivity rate of environmental screening samples was 19.0% (73/385). Highest percentages were found in the shower drain samples (38.2%) and the toilet water samples (25.0%). Sink drain samples showed least CPE positivity (3.3%). The WGS data revealed long-term co-existence of three patient sample derived C. freundii clusters. The biggest cluster (ST22) connects 12 patients and 8 environmental isolates taken between 2018 and 2021 spread across the ward. In an overlapping period, another cluster (ST170) links eight patients and four toilet water isolates connected to the same room. The third C. freundii cluster (ST421) connects two patients hospitalised in the same room but over a period of one and a half year. Additional sampling in 2022 revealed clonal isolates linked to the two largest clusters (ST22, ST170) in the wastewater collection pipes connecting the rooms. Conclusions Our findings suggest long-term circulation and transmission of carbapenemase producing C. freundii clones in hospital sanitary installations despite surveillance, daily cleaning and intermittent disinfection protocols. We propose a role for the wastewater drainage system in the spread within and between rooms and for the sanitary installations in the indirect transmission via bioaerosol plumes. To tackle this problem, a multidisciplinary approach is necessary including careful design and maintenance of the plumbing system.

Objectives: The aim of this systematic review was to investigate the effect of antibiotics on the eradication of multidrug-resistant organisms (MRO) in intestinal carriers. We defined multidrug-resistant organisms as vancomycin-resistant Enterococcus faecium (VREfm), and multidrug-resistant Gram-negative Enterobacterales. Methods: We searched the EMBASE, Cochrane Central, and PubMed databases from inception to medio November 2023. We included randomised and controlled clinical trials (RCTs), that investigated the effect of antibiotics on the eradication of multidrug-resistant organisms in intestinal carriers. Finally, we performed a meta-analysis. Results: We included five RTCs in the systematic review. In four studies an effect of antibiotics on the eradication of MRO was shown at the end of intervention, but it was not sustained at follow-up. In the fifth study, the effect at the end of intervention was not reported, and there was no observed effect of the intervention at follow-up. We included four studies in the meta-analysis, and it suggests an effect of antibiotics on the eradication of MRO in intestinal carriers at the end of follow-up with a p-value of 0.04 (95% confidence interval 1.02–1.95). None of the studies reported a significant increase in resistance to the study drug. Gastrointestinal disorders were the most frequent non-severe adverse event. Conclusions: The effect of antibiotics on the eradication of multidrug-resistant organisms in intestinal carriers was not statistically significant in any of the five included studies; however, we found a significant effect in the pooled meta-analysis. As the confidence interval is large, we cannot determine the clinical importance of this finding, and it should be further investigated.

ABSTRACTRapid and reliable detection of carbapenemase-producing Enterobacterales (CPE) is crucial for prompt treatment and infection control. Most assays target the primary four enzymes (KPC, OXA-48-like, VIM, and NDM), often missing less common variants (e.g., GES, IMI, OXA-23, and OXA-58). Therefore, assays based on the hydrolysis of carbapenems are recommended in addition to differentiation tests such as PCR or immunochromatographic assays. The aim of this study was to compare the currently Clinical and Laboratory Standards Institute (CLSI)-recommended tests mCIM (modified carbapenem inactivation method) and Carba NP with new colorimetric tests (NitroSpeed-Carba NP) and novel variations of the carbapenem inactivation method (CIM) such as simplified CIM (sCIM) or modified zinc-supplemented CIM (mzCIM). The challenge collection included 205 clinical isolates, 139 CPE vs 66 non-CPE. Among all 205 isolates, the sensitivity/specificity of mCIM was 81.3%/98.5%, Carba NP 76.3%/100%, NitroSpeed-Carba NP 86.3%/78.8%, sCIM 100%/94%, and mzCIM 97.8%/98.5%. For rare carbapenemases (n = 48), the sensitivity of mzCIM (98.3%) and sCIM (100%) was higher than that of mCIM (60.4%), Carba NP (50%), or NitroSpeed-Carba NP (70.2%). Most indeterminate results occurred for mCIM (14.4%), Carba NP (8.2%), and sCIM (6.3%). The detection of rare carbapenemases remains challenging with the currently recommended assays. The CIM-based tests demonstrated superior sensitivity, with sCIM and mzCIM outperforming the currently recommended mCIM and Carba NP, especially among isolates with weakly hydrolyzing carbapenemases (e.g., OXA-23 and OXA-58). Although colorimetric assays provide more rapid results, laboratories have to be aware of the low sensitivity for rare carbapenemases. Both sCIM and the new mzCIM performed well, are cost-effective, and can easily be implemented in any laboratory.IMPORTANCEDetection of so-called rare carbapenemases (e.g., GES, IMI, OXA-23, and OXA-58) in Enterobacterales is challenging, and data on the performance of currently available assays are scarce. This study systematically assessed the performance of currently recommended and novel hydrolysis-based assays on a set of molecularly characterized isolates. It demonstrates that the currently recommended assays mCIM and Carba NP perform well on isolates producing common carbapenemases such as KPC, VIM, NDM, and OXA-48, but have only a moderate sensitivity in the detection of rare carbapenemases. In contrast, the newer CIM-based variants, sCIM and mzCIM, are equally capable of detecting frequent and uncommon carbapenemases. These assays could potentially help to improve our knowledge on the epidemiology of these “rare” enzymes.

The emergence of extended-spectrum β-lactamase and carbapenemase-producing Enterobacterales (ESBL-E and CPE, respectively) is a threat to modern medicine, as infections become increasingly difficult to treat. These bacteria have been detected in aquatic environments, which raises concerns about the potential spread of antibiotic resistance through water. Therefore, we investigated the occurrence of ESBL-E and CPE in surface water in Lower Saxony, Germany, using phenotypic and genotypic methods. Water samples were collected from two rivers, five water canals near farms, and 18 swimming lakes. ESBL-E and CPE were isolated from these samples using filters and selective agars. All isolates were analyzed by whole genome sequencing. Multidrug-resistant Enterobacterales were detected in 4/25 (16%) water bodies, including 1/2 rivers, 2/5 water canals and 1/18 lakes. Among all samples, isolates belonging to five different species/species complexes were detected: Escherichia coli (n = 10), Enterobacter cloacae complex (n = 4), Citrobacter freundii (n = 3), Citrobacter braakii (n = 2), and Klebsiella pneumoniae (n = 2). Of the 21 isolates, 13 (62%) were resistant at least to 3rd generation cephalosporins and eight (38%) additionally to carbapenems. CPE isolates harbored blaKPC-2 (n = 5), blaKPC-2 and blaVIM-1 (n = 2), or blaOXA-181 (n = 1); additionally, mcr-9 was detected in one isolate. Two out of eight CPE isolates were resistant to cefiderocol and two to colistin. Resistance to 3rd generation cephalosporins was mediated by ESBL (n = 10) or AmpC (n = 3). The presence of AmpC-producing Enterobacterales, ESBL-E and CPE in northern German surface water samples is alarming and highlights the importance of aquatic environments as a potential source of MDR bacteria.

A rapid phenotypic carbapenemase-producing Enterobacterales (CPE) detection method was established using Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS). Sixtyfour carbapenem-resistant Enterobacterales (CRE) strains from Samutprakan Hospital were examined using MALDI-TOF MS by the ertapenem hydrolysis method and the susceptibility was compared with a modified carbapenem inactivation method (mCIM). Drug resistance genes were detected by polymerase chain reaction (PCR). The ertapenem drug and bacterial strains were mixed and drug hydrolysis owing to CPE activity was confirmed by specific molecular masses of ertapenem [M+H]+ at 476.5 m/z (±500 ppm), with peak disappearance judged as carbapenemase-positive. The most common CRE species were Klebsiella pneumoniae, Escherichia coli, and Enterobacter cloacae. From 58 CPE strains, 17 strains of K. pneumoniae (29.3%) harbored blaNDM and blaOXA- 48-like genes together, while 33 strains of K. pneumoniae (56.9%), 6 strains of E. coli (10.3%) and 1 strain of Ent. cloacae (1.75%) carried blaNDM or blaOXA-48- like genes alone and 1 strain of K. pneumoniae (1.75%) contained blaKPC. After 3 h of incubation with ertapenem, all 58 drug-resistant strains revealed disappearance of the ertapenem-specific waveform peak at 476.5 m/z, whereas 6 strains of CRE (non-CPE) revealed the ertapenemspecific waveform peak. The MALDI-TOF MS and mCIM data were 100% consistent. The MALDI-TOF MS based ertapenem hydrolysis assay was demonstrated as a rapid and accurate method to detect carbapenemase activity of Enterobacterales strains that can be routinely performed in clinical microbiology laboratories. [ABSTRACT FROM AUTHOR]

The Carnian Pluvial Episode (CPE), a 1–2 Ma interval of enhanced humidity midway through the Triassic Period, has been implicated in high levels of biotic turnover in marine environments. The Carnian stage on land also saw major faunal and floral reorganisation, including the extinction of rhynchosaurs, the diversification of dinosaurs and the origin of crocodylomorphs and mammaliaforms. An imperfect fossil record and dating uncertainties, however, make establishing a firm link to the CPE much more difficult, but the picture is gradually improving as new evidence accumulates. We review observed terrestrial biotic changes and suggest that in most cases the effects of the CPE in isolation remain ambiguous or were relatively minor or reversible. Rather, the Carnian biotic change was probably a staggered response to a number of factors, including climatic, set against the backdrop of long-term rebuilding of ecosystems following the end-Permian mass extinction. [ABSTRACT FROM AUTHOR]

Complete panicle exsertion (CPE) in rice is an important determinant of yield and a desirable trait in breeding. However, the genetic basis of CPE in rice still remains to be completely characterized. An ethyl methane sulfonate (EMS) mutant line of an elite cultivar Samba Mahsuri (BPT 5204), displaying stable and consistent CPE, was identified and named as CPE-110. MutMap and RNA-seq were deployed for unraveling the genomic regions, genes, and markers associated with CPE. Two major genomic intervals, on chromosome 8 (25668481-25750456) and on chromosome 11 (20147154-20190400), were identified to be linked to CPE through MutMap. A non-synonymous SNP (G/A; Chr8:25683828) in the gene LOC_Os08g40570 encoding pyridoxamine 5′-phosphate oxidase with the SNP index 1 was converted to Kompetitive allele-specific PCR (KASP) marker. This SNP (KASP 8-1) exhibited significant association with CPE and further validated through assay in the F2 mapping population, released varieties and CPE exhibiting BPT 5204 mutant lines. RNA-seq of the flag leaves at the booting stage, 1100 genes were upregulated and 1305 downregulated differentially in CPE-110 and BPT 5204. Metabolic pathway analysis indicated an enrichment of genes involved in photosynthesis, glyoxylate, dicarboxylate, porphyrin, pyruvate, chlorophyll, carotenoid, and carbon metabolism. Further molecular and functional studies of the candidate genes could reveal the mechanistic aspects of CPE. [ABSTRACT FROM AUTHOR]

Infection prevention and control (IPC) practices vary among companion animal clinics, and outbreaks with carbapenemase-producing Enterobacterales (CPE) have been described. To investigate the effect of an IPC intervention (introduction of IPC protocols, IPC lectures, hand hygiene campaign) in four companion animal clinics. IPC practices, environmental and hand contamination with antimicrobial-resistant micro-organisms (ARM) and hand hygiene (HH) were assessed at baseline, and 1 and 5 months after the intervention. Median IPC scores (% maximum score) improved from 57.8% (range 48.0–59.8%) to 82.9% (range 81.4–86.3%) at 1-month follow-up. Median cleaning frequency assessed by fluorescent tagging increased from 16.7% (range 8.9–18.9%) to 30.6% (range 27.8–52.2%) at 1-month follow-up and 32.8% (range 32.2–33.3%) at 5-month follow-up. ARM contamination was low in three clinics at baseline and undetectable after the intervention. One clinic showed extensive contamination with ARM including CPE before and after the intervention (7.5–16.0% ARM-positive samples and 5.0–11.5% CPE-positive samples). Mean HH compliance improved from 20.9% [95% confidence interval (CI) 19.2–22.8%] to 42.5% (95% CI 40.4–44.7%) at 1-month follow-up and 38.7% (95% CI 35.7–41.7%) at 5-month follow-up. Compliance was lowest in the pre-operative preparation area at baseline (11.8%, 95% CI 9.3–14.8%) and in the intensive care unit after the intervention (28.8%, 95% CI 23.3–35.1%). HH compliance was similar in veterinarians (21.5%, 95% CI 19.0–24.3%) and nurses (20.2%, 95% CI 17.9–22.7%) at baseline, but was higher in veterinarians (46.0%, 95% CI 42.9–49.1%) than nurses (39.0%, 95% CI 36.0–42.1%) at 1-month follow-up. The IPC intervention improved IPC scores, cleaning frequency and HH compliance in all clinics. Adapted approaches may be needed in outbreak situations. [ABSTRACT FROM AUTHOR]

Background: Between January and September 2022, 10 patients who were admitted to the Intensive Care Unit (ICU) for a period of more than 72 hours were found to have carbapenemase-producing Enterobacteriaceae (CPE) in their screening/clinical specimens. No epidemiological association was found among them, and none had received healthcare outside Canada in the preceding 12 months. The study investigated whether sink drains could be a source of CPE transmission. Methods: A retrospective study was conducted on all patients admitted to the ICU during the surveillance period who tested positive for CPE, alongside a pilot study involving environmental screening cultures collected from hand hygiene (HH) and washroom sink drains in ICU rooms after the patients had been discharged. Information was gathered on CPE risk factors, such as recent travel history or hospitalization outside of Canada, endoscopic retrograde cholangiopancreatography procedures, or hemodialysis within the last 12 months. Results: During the surveillance period, 15 non-duplicate CPE isolates were obtained from 14 patients in the ICU. Out of the 14, 10 patients were identified after ICU stays of greater than 72 hours. Klebsiella pneumoniae carbapenemase (KPC) was the most identified carbapenemase, found in five (50%) of isolates, followed by three (30%) New Delhi metallo-β-lactamase, and two (20%) Oxacillinase-48 like carbapenemase. In addition, a total of 33 sink drains underwent testing: 24 were HH sinks, while nine were inpatient washrooms. Among the 24 HH sink drains, nine (37.5%) tested positive for CPE, eight of these sink drains (88.9%) had KPC genes, two (22%) had NDM genes, and one (11%) had VIM genes. Among the HH sink drains contaminated by CPE, three (33.3%) out of these nine shared the same species/gene combination (KPC-Citrobacter freundii) as the CPE-positive patient recently discharged from the room. Conclusion: These findings suggest a potential link between KPC-producing Citrobacter freundii detected in the HH sink drain and a CPE-positive patient. However, the direction of transmission remains unclear. Further research is necessary to analyze the molecular genotyping and validate any potential relatedness among them, in addition, collaborative efforts from the infection prevention and control, microbiology, environmental, and facilities teams are essential to eliminate CPE from HH sink drains. [ABSTRACT FROM AUTHOR]

Background: The evolution and springing up of carbapenemase-producingEnterobacteriaceae (CPE) threatens worldwide health. The capability of carbapenemases to degrade all β-lactam antibiotics leads to less antibiotics retaining activity against infections caused by CPE that are associated with high mortality and bad prognosis. There is an urgent need to define robust standardized screening methods for the effective detection of CPE in order to control their spread. This study aimed to detect the efficiency of Chromatic TM CPE medium in identifying CRE directly from ICU clinical samples. Methods: A Cross section study was conducted at intensive care units and Medical Microbiology and Immunology laboratory, Faculty of medicine, Ain Shams university. The sample was directly inoculated on CRE chromagar plate and incubated at 35°C aerobically. Isolation and identification of CPE was performed according to Manual of Clinical Microbiology 2019 then antimicrobial susceptibility test by Kirby-Baur disc diffusion. The sensitivity and specificity of CRE screening by CRE chromogenic media were detected using Modified carbapenemase inactivation method (mCIM) as the standard method. Results: One hundred and fifty-eight isolates were obtained from hospitalized ICU patients in Ain shams university hospital between April to November 2022. There were different types of samples, the most common one was sputum. Among 71 isolates grew on blood and MacConkey agar media, 59 (83.15%) showed growth on chromogenic media while 12 (16.9%) showed no growth on chromogenic media. Fifty-four (76.1%) showed MDR Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii. Among 54 MDR isolates, 50 (92.6%) were positive carbapenemase production, while 4 (7.4%) were negative carbapenemase production using mCIM test as confirmatory test. The sensitivity of chromogenic media was 98%, specificity was 50% in all isolates and the accuracy of the test was 94.4%. While the sensitivity of chromogenic media in all Enterobacteriaceae was 96.9%, specificity was 33.3%. Conclusion: This study revealed that CRE chromogenic is highly sensitive in the screening of CRE detection. It efficiently saves more time in identifying CRE as compared to mCIM method. The use of a single chromogenic medium can reduce the cost of sample processing and be an effective tool for rapid detection of CRE. Accurate and fast detection of patients colonized by CPEs is clinically important in applying proper infection control precautions.

The present paper deals with the development of a new and original hexadecyltrimethylammonium bromide (CTAB)-modified carbon paste electrode (CPE) dedicated for the direct analysis of nitrites in aquaculture and rat blood. The surface morphologies and contact angles of unmodified and modified CPEs were performed using Scanning Electron Microscopy (SEM), Optical Microscopy (OM) and sessile drop techniques, respectively. Cyclic Voltammetry (CV) was used to study the electrochemical behaviour of nitrites, whilst Differential Pulse Voltammetry (DPV) and Linear Sweep Voltammetry (LSV) techniques were used to investigate their quantitative analysis. DPV operating parameters were optimized and the optimum values were pulse height 0.4 V, scan increment 0.015 V, pulse width 0.01 s and step time 0.5 s. The repeatability and reproducibility of the CPE were studied, and the effect of some interfering ions was evaluated to highlight the selectivity of the electrode. The modified CTAB/CPE showed a good stability and proved to be more sensitive, leading to a Limit of Detection (LOD) of 0.034 mg L−1 and a Limit of Quantification (LQ) of 0.152 mg L−1. The elaborated sensors were applied with success for nitrites analysis in aquaria with CPE and in rat blood with CTAB/CPE, preparing human blood measurements in near future work. [ABSTRACT FROM AUTHOR]

Background: Accumulating evidence shows a role of the hospital wastewater system in the spread of multidrug-resistant organisms, such as carbapenemase producing Enterobacterales (CPE). Several sequential outbreaks of CPE on the geriatric ward of the Ghent University hospital have led to an outbreak investigation. Focusing on OXA-48 producing Citrobacter freundii, the most prevalent species, we aimed to track clonal relatedness using whole genome sequencing (WGS). By exploring transmission routes we wanted to improve understanding and (re)introduce targeted preventive measures. Methods: Environmental screening (toilet water, sink and shower drains) was performed between 2017 and 2021. A retrospective selection was made of 53 Citrobacter freundii screening isolates (30 patients and 23 environmental samples). DNA from frozen bacterial isolates was extracted and prepped for shotgun WGS. Core genome multilocus sequence typing was performed with an in-house developed scheme using 3,004 loci. Results: The CPE positivity rate of environmental screening samples was 19.0% (73/385). Highest percentages were found in the shower drain samples (38.2%) and the toilet water samples (25.0%). Sink drain samples showed least CPE positivity (3.3%). The WGS data revealed long-term co-existence of three patient sample derived C. freundii clusters. The biggest cluster (ST22) connects 12 patients and 8 environmental isolates taken between 2018 and 2021 spread across the ward. In an overlapping period, another cluster (ST170) links eight patients and four toilet water isolates connected to the same room. The third C. freundii cluster (ST421) connects two patients hospitalised in the same room but over a period of one and a half year. Additional sampling in 2022 revealed clonal isolates linked to the two largest clusters (ST22, ST170) in the wastewater collection pipes connecting the rooms. Conclusions: Our findings suggest long-term circulation and transmission of carbapenemase producing C. freundii clones in hospital sanitary installations despite surveillance, daily cleaning and intermittent disinfection protocols. We propose a role for the wastewater drainage system in the spread within and between rooms and for the sanitary installations in the indirect transmission via bioaerosol plumes. To tackle this problem, a multidisciplinary approach is necessary including careful design and maintenance of the plumbing system. [ABSTRACT FROM AUTHOR]

Clostridium perfringens enterotoxin (Cpe)-producing strains cause gastrointestinal infections in humans and account for the second-largest number of all foodborne outbreaks caused by bacterial toxins. The Cpe toxin is only produced during sporulation; this process might be affected when C. perfringens comes into contact with host cells. The current study determined how the cpe expression levels and spore formation changed over time during co-culture with Caco-2 cells (as a model of intestinal epithelial cells). In co-culture with Caco-2 cells, total C. perfringens cell counts first decreased and then remained more or less stable, whereas spore counts were stable over the whole incubation period. The cpe mRNA level in the co-culture with Caco-2 cells increased more rapidly than in the absence of Caco-2 cells (3.9-fold higher levels in coculture than in the absence of Caco-2 cells after 8 h of incubation). Finally, we found that cpe expression is inhibited by a cue released by Caco-2 cells (8.3-fold lower levels in the presence of supernatants of Caco-2 cells than in the absence of the supernatants after 10 h of incubation); as a consequence, the increased expression in co-culture with Caco-2 cells must be caused by a factor associated with the Caco-2 cells.

Antimicrobial resistance (AMR) is a worldwide healthcare problem. Multidrug-resistant organisms (MDROs) can spread quickly owing to their resistance mechanisms. Although colonized individuals are crucial for MDRO dissemination, colonizing microbes can lead to symptomatic infections in carriers. Carbapenemase-producing Enterobacterales (CPE) are among the most important MDROs involved in colonizations and infections with severe outcomes. This review aimed to track down the first reports of CPE in Africa, describe their dissemination throughout African countries and summarize the current status of CRE and CPE data, highlighting current knowledge and limitations of reported data. Two database queries were undertaken using Medical Subject Headings (MeSH), employing relevant keywords to identify articles that had as their topics beta-lactamases, carbapenemases and carbapenem resistance pertaining to Africa or African regions and countries. The first information on CPE could be traced back to the mid-2000s, but data for many African countries were established after 2015–2018. Information is presented chronologically for each country. Although no clear conclusions could be drawn for some countries, it was observed that CPE infections and colonizations are present in most African countries and that carbapenem-resistance levels are rising. The most common CPE involved are Klebsiella pneumoniae and Escherichia coli, and the most prevalent carbapenemases are NDM-type and OXA-48-type enzymes. Prophylactic measures, such as screening, are required to combat this phenomenon.