European Committee Research Articles

Susceptibility of various Gram-negative bacteria to antibacterial agents: SMART in China 2019–2020

BackgroundThe Study for Monitoring Antimicrobial Resistance Trends (SMART) is an international surveillance program longitudinally monitoring aerobic and facultative Gram-negative bacteria (GNB) involvement in infections and their antimicrobial resistance profiles. Here the incidence and resistance patterns of Chinese GNB isolates from bloodstream infections (BSI), intraabdominal infections (IAI), respiratory tract infections (RTI) and urinary tract infections (UTI) to commonly used antibacterial agents has been updated. 4,975 GNB isolates collected from 22 hospitals across 7 regions of China from 2019 to 2020 were analyzed. Antimicrobial minimum inhibitory concentrations were assessed using broth microdilution, and susceptibility interpretations followed the breakpoints of European Committee on Antimicrobial Susceptibility Testing 2022 or Clinical and Laboratory Standards Institute.ResultsThis study affirmed that Escherichia coli (Ec) was the most commonly identified GNB (32.1%) and then Klebsiella pneumoniae (Kp) (25.3%), Pseudomonas aeruginosa (Pa) (13.9%) and Acinetobacter baumannii (10.5%). The detection rates of carbapenem-resistant (CR) Enterobacterales varied across major infection sites, ranging from 10.3% in UTI to 18.9% in RTI. Specifically, the detection rates of CR-Kp and CR-Pa ranged from 16.2% in IAI to 35.8% in UTI and from 16.1% in UTI to 38.0% in RTI, respectively. Extended-spectrum β-lactamases (ESBL)-producing Ec and Kp bacteria exhibited over 91.7% susceptibility to carbapenems and at least 87.8% susceptibility to amikacin and colistin, but showed lower susceptibility to piperacillin/tazobactam (57.5–86.2%), levofloxacin (10.8–39.7%) and aztreonam (15.3–27.6%) across different infection sources. Amikacin showed higher efficacy against CR strains compared to other commonly used antibacterial drugs, with 80.0% susceptibility against CR-Ec and 82.3% susceptibility against CR-Pa, while only 36.3% susceptibility was observed against CR-Kp.ConclusionsThe study found varying incidences of CR isolates in Chinese hospitals. Treatment options remained limited due to resistance to multiple antibacterial agents. Carbapenems demonstrated effective antimicrobial in vitro activity against ESBL-producing Enterobacterales found in BSI, IAI, UTI and RTI, outperforming broad-spectrum cephalosporins and other β-lactamase inhibitors.

Performance of a broth microdilution assay for routine minimum inhibitory concentration determination of 14 anti-tuberculous drugs against the Mycobacterium tuberculosis complex based on the EUCAST reference protocol.

This comparative study aimed at qualifying a broth microdilution (BMD) assay for phenotypic drug susceptibility testing (pDST) of Mycobacterium tuberculosis complex (MTBC) strains for implementation in a routine DST workflow. The assay was developed based on the EUCAST (European Committee on Antimicrobial Susceptibility Testing) reference protocol for determination of the minimum inhibitory concentration (MIC) of 14 anti-tuberculous drugs (isoniazid [INH], rifampicin [RIF], ethambutol [EMB], amikacin [AMI], moxifloxacin [MFX], levofloxacin [LFX], bedaquiline [BDQ], clofazimine [CFZ], delamanid [DLM], pretomanid [PA], para-aminosalicylic acid [PAS], linezolid [LZD], ethionamide [ETH], and cycloserine [CS]). Forty MTBC strains with various drug resistance profiles were tested to determine the agreement between MIC results and genotypic drug susceptibility testing (gDST) results derived from whole-genome sequencing (WGS). The agreement between the BMD and gDST results was solid for the majority of the drugs (average agreement 98%, range 90%-100%), including key drugs such as INH, RIF, MFX, LFX, BDQ, DLM, and PA. Ten discrepancies were identified (corresponding to 1.8% of the total number of MIC determinations) and most (8/10) were characterized by MICs equal or close to the potential critical concentration (pCC) applied in the BMD assay. Importantly, the assay can be adjusted to new drug recommendations and concentrations, tailored to local needs. We conclude that the BMD assay provides reliable results, and its implementation in our MTBC routine workflow will produce valuable data that improve our understanding and management of MTBC drug resistance.

Temporal trends in antimicrobial resistance of medically important pathogens on Curaçao

BackgroundThe Caribbean lacks recent comprehensive antimicrobial resistance data to inform clinicians and decision-makers. This study aims to provide a snapshot of susceptibility trends for pathogens on Curaçao, an island in the southern Caribbean.MethodsWe analyzed susceptibility data of bacterial pathogens isolated from samples submitted from patients attending general practitioners, outpatient clinics and those who were hospitalized between January 2018 and December 2023. Samples originating from blood, urine, genital tract, soft tissue, and lungs were included. Susceptibility testing was performed by VITEK2 according using the European Committee on Antimicrobial Susceptibility Testing criteria.ResultsIn total, 13,528 patients contributed to 22,876 first isolates. Of all infections in adults with Staphylococcus aureus, 14% (95% confidence interval 10–18%) were methicillin-resistant S. aureus (MRSA) in blood, and up to 27% (20–35%) in soft-tissue cultures. For Escherichia coli and Klebsiella pneumoniae, resistance levels were up to 27% (95% CI 22–32%) for cefuroxime and up to 18% (95% CI 18–28%) for third-generation cephalosporins. The addition of gentamicin to empirical therapy with cefuroxime increased coverage only moderately, especially in K. pneumoniae (16%, 95% CI 12–20%). Resistance to amoxicillin-clavulanic acid was up to 47% (95% CI 43–50%), to ciprofloxacin 34% (95% CI 31–37%) and to cotrimoxazole 37% (95% CI 34–39) in urine cultures from outpatients. In contrast, low levels of carbapenem resistant Enterobacterales and Pseudomonas aeruginosa were observed.ConclusionsAntimicrobial resistance is high and widespread across several important antibiotic classes. The widespread occurrence of MRSA and resistance to third-generation cephalosporins highlights the importance of identifying risk factors, enabling more effective guidance for antimicrobial stewardship.

Australian Group on Antimicrobial Resistance (AGAR) Australian Staphylococcus aureus Surveillance Outcome Program (ASSOP) Bloodstream Infection Annual Report 2023.

From 1 January to 31 December 2023, fifty-seven institutions across Australia participated in the Australian Staphylococcus aureus Surveillance Outcome Program (ASSOP). The aim of ASSOP 2023 was to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on methicillin resistance, and to characterise the methicillin-resistant S. aureus (MRSA) molecular epidemiology. A total of 3,422 SAB episodes were reported, of which 77.0% were community-onset. Overall, 16.1% of S, aureus were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 14.8%, which was not significantly different to the 16.5% all-cause mortality associated with methicillin-susceptible SAB (p = 0.44). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S, aureus (MSSA) was infrequent. However, in addition to the β-lactams, approximately 33% of MRSA were resistant to ciprofloxacin; 30% to erythromycin; 13% to tetracycline; 13% to gentamicin; and 3% to co-trimoxazole. Two New South Wales daptomycin-resistant MRSA, with minimum inhibitory concentrations (MICs) of 3.0 and 4.0 mg/L, were identified as ST22-IV, with a V351E mprF mutation, and ST45-V with a T345I mprF mutation respectively. Three daptomycin-resistant MSSA were identified. One from Tasmania, with a daptomycin MIC of 1.5 mg/L, identified as ST9295 with a L341I mprF mutation; one from New South Wales, with a daptomycin MIC of 3.0 mg/L, identified as ST97 with a L776S mprF mutation; and one from Western Australia, with a daptomycin MIC of 2.0 mg/L, identified as ST5. No previously reported mutations in known loci were detected in the Western Australian isolate. When applying the European Committee on Antimicrobial Susceptibility Testing breakpoints, teicoplanin resistance was detected in three MSSA isolates and one MRSA isolate. Vancomycin or linezolid resistance was not detected. Resistance to non-β-lactam antimicrobials was largely attributable to the healthcare-associated MRSA (HA-MRSA) clone ST22-IV [2B] (EMRSA-15), and the community-associated MRSA (CA-MRSA) clone ST45-V [5C2&5] which has acquired resistance to multiple antimicrobials including ciprofloxacin, clindamycin, erythromycin, gentamicin, and tetracycline. ST22-IV [2B] (EMRSA-15) was the predominant HA-MRSA clone in Australia. Overall, 85% of methicillin-resistant SAB were caused by community-associated MRSA (CA-MRSA) clones. Although polyclonal, approximately 70.3% of CA-MRSA clones were characterised as ST93-IV [2B] (Queensland clone); ST5-IV [2B]; ST1-IV [2B]; ST45-V [5C2&5]; ST30-IV [2B]; ST8-IV [2B]; ST6-IV [2B]; ST97-IV [2B]; and ST953-IV [2B]. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S, aureus bacteraemia.

A potential antifungal bioproduct for Microsporum canis: Bee venom.

Natural treatment options forMicrosporum canisdermatophytosis are being explored because of resistance to several antifungal medications. In this study, the potential antifungal effect of bee venom (BV), a natural antimicrobial agent, onM. caniswas investigated. The antifungal effects of BV, fluconazole, itraconazole, amphotericin B and terbinafine were evaluated by the macrodilution method at various concentrations by modifying the microdilution method recommended by the European Committee on Antimicrobial Susceptibility Testing. All isolates were observed to be susceptible to terbinafine and fully resistant to fluconazole and amphotericin B. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values ofM. canisisolate 2 (Mc2) were determined as 8 µg/mL for itraconazole. The MIC and MFC values of BV were found to be 320 µg/mL for the Mc2 isolate and 640 µg/mL for the Mc6 isolate. The results showed that the isolates obtained from clinical samples in this study were highly resistant to all antifungal agents, except terbinafine. The increase in resistance indicates that antifungal drugs will become insufficient and ineffective over time and natural products such as BV should be evaluated as alternatives.Contribution:Although there are many drugs for the treatment ofM. canis, the increase in resistance to antifungal agents reveals the need for the identification and development of new natural agents. Bee venom, which has been shown to have a safe and weak allergenic effect in various studies, can be tested for usability as a local antifungal drug when supported byin vivostudies.

WAYS OF IMPLEMENTING A COMPLEX APPROACH TO INCREASE THE COMPETITIVENESS OF UKRAINE

The study focuses on examining the key factors influencing the competitive state of Ukraine’s socio-economic model at the current stage of development. An analysis of major macroeconomic parameters on the eve of Russia’s full-scale invasion revealed substantial potential for economic recovery following the losses incurred due to COVID-19 pandemic restrictions. Taking into account recent European Union reforms aimed at enhancing competitiveness based on the European Commission’s Communication to the European Parliament, the Council, the European Economic and Social Committee, and the Committee of the Regions, titled «Long-term competitiveness of the EU: looking beyond 2030,» certain related directions for implementing relevant transformations in Ukraine have been identified. As a result, the authors developed a comprehensive approach to enhance the competitiveness of Ukraine. Correspondingly, possible pathways for implementing specific components of this comprehensive approach were proposed. It has been determined that, through incentives involving public and business influence on the elite, it is necessary to ensure the development, implementation, and monitoring of the most urgent reforms, namely: security reform, institutional reform, infrastructure reconstruction, and innovation reform. The most critical steps were identified, primarily involving the deepening of cooperation with foreign partners at both business and governmental levels. The need for decisive action against corruption in all areas of life and the urgent institutional reform required was also highlighted. Using G. Hofstede’s cultural dimensions theory, a regression analysis was conducted to explore the relationship between institutional quality ratings from the “Global Competitiveness Index” and Ukraine’s cultural dimension values. Based on these calculations and the evaluation of statistical correlation density through the determination coefficient, it was found that higher individualism correlates with higher institutional quality, while higher power distance correlates with lower institutional quality. The dependency of institutions on other cultural dimensions was too weak to indicate a meaningful relationship; however, graphical analysis of long-term orientation and uncertainty avoidance suggested a shared dynamic. Three main directions for implementing institutional reform were proposed: fostering civic patriotism among the population, combating the old, corrupt, anti-national elite (oligarchs), and cultivating patriotic meritocrats as replacements. Meeting these conditions would establish a strong foundation for further development of a rule-of-law state. Regarding infrastructure reconstruction and innovation reform, plans for modernization and digitalization were proposed. Particular attention should be paid to implementing and utilizing modern tools to leverage the full spectrum of financial and stock market opportunities. The implementation of modern educational approaches at all levels is also of great importance. The successful combination and implementation of these proposed processes will lead to increased productivity and competitiveness. The driver of positive changes should be Ukrainian business and civil society: entrepreneurs understand what is needed for dynamic growth, and citizens know what is required for a decent life.

Ozanimod in Relapsing Multiple Sclerosis: New Insights

Ozanimod is an approved treatment for relapsing forms of multiple sclerosis (RMS) that has been shown to reduce relapses, new brain lesions, and brain volume loss relative to intramuscular interferon (IFN) β-1a. This article summarizes selected data from clinical trials of ozanimod in RMS, which were presented at the 40th Congress of the European Committee for Treatment and Research in Multiple Sclerosis (ECTRIMS) held in Denmark in September 2024 and the Annual Meeting of the European Charcot Foundation held in Italy in November 2024. Final safety data were presented for the completed open-label extension (OLE) study of ozanimod in adults with RMS (DAYBREAK), showing that rates of adverse events (AE) remained stable or decreased over 8 years of ozanimod treatment. These findings confirm the established safety profile of ozanimod. An analysis of brain volume changes across DAYBREAK and parent trials were also presented, demonstrating that the annualized rate of brain volume loss in participants treated with continuous ozanimod for up to 7 years was below the pathologic cutoff and was similar to those previously reported from healthy controls. Data were presented from a retrospective, observational study into the effects of switching from fingolimod to ozanimod in patients with RMS due to safety or intolerance reasons. The authors concluded that this study confirms that switching from fingolimod to ozanimod for safety reasons such as lymphopenia or hypertransaminasemia may be a good strategy to continue sphingosine-1-phosphate receptor (S1PR)-modulating drug therapy. Finally, an ad hoc interim analysis of patients with early RMS from ENLIGHTEN showed promising results, demonstrating that more than three-quarters of patients had either stable or improved cognitive processing speed after 1 year of ozanimod 0.92 mg.

Utilizing an Opportunistic Clinical Study and Population-Based Pharmacokinetic Models to Identify Rational Empiric Dosing Regimens for Piperacillin-Tazobactam in Critically Ill Patients.

Determining an effective dosing regimen for piperacillin-tazobactam in critically ill patients is challenging due to substantial pharmacokinetic variability caused by complex pathophysiological changes. To address this need, a prospective clinical study was conducted, which enrolled 112 critically ill patients and employed an opportunistic sampling strategy. Population modeling and simulation were performed to characterize the pharmacokinetics (PK) and probability of target attainment (PTA) of piperacillin-tazobactam under various dosing regimens. Both piperacillin and tazobactam final models were one-compartment models with zero-order input and first-order elimination. Significant covariates included lean body weight for piperacillin and creatinine clearance along with continuous renal replacement therapy (CRRT) for both drugs. Monte Carlo simulations demonstrated that continuous infusion can achieve higher PTA than intermittent and extended infusions. When considering the minimum inhibitory concentration (MIC) of 16 mg/L for Pseudomonas aeruginosa (a frequently encountered bacterial pathogen among critically ill patients) and a PK/PD target of 100% fT >MIC, continuous infusion of 6 g/day is recommended for critically ill patients with a CLcr <60 mL/min, 9 g/day for patients with CLcr in the range of 60 to 129 mL/min, and 12 g/day for patients with a CLcr ≥130 mL/min. In addition, extended infusion represents a good alternative, especially the 3 g q6h or 4 g q6h regimens which can achieve the designated European Committee on Antimicrobial Susceptibility Testing (EUCAST) non-species-related PK/PD breakpoint of 8 mg/L. Our study provided valuable insight into PTA outcomes, which, together with individual renal function of future patients and institution-specific piperacillin susceptibility patterns, may assist physicians when making dosing decisions.

A Study Investigating The Presence Of Diatoms On Tooth Samples Using The Acid Digestion Method For The Estimation Of Time Since Death In Drowning Cases

Teeth are a further source of diatoms in establishing the cause of death and time since death by analysis of the stage qualitative and quantitative evaluation of diatoms diversity. Trials also confirmed that diatoms have also been a useful tool in differentiating antemortem and postmortem drowning (Jian Zhao April 2014). Diatoms are a type of algae that belong to the phylum Bacillariophyta. They are unicellular organisms with a unique feature – their cell walls are made of silica, which gives them a glass-like appearance. Diatoms are a diverse group of microorganisms found in various aquatic environments, including oceans, lakes, rivers, and even damp soil. This study mainly focuses on a tooth immersed in a water sample from a known water body for a day and the 7th day to observe the variation and the stages of the life cycle. The method of extraction used is the acid digestion method using concentrated Sulphuric Acid. We could find the centric and pennate types of diatoms. Diatoms are also important as they indicate natural water quality (European Committee of Standardization 2004). According to the results, we cannot deny that teeth are a source for the extraction of diatoms and also an indicator of time since death when other methods fail.

Molecular Identification of OXA Carbapenemase-Encoding Genes in Acinetobacter baumannii Isolated from Patients in Critical Care in Egypt.

Background: The emergence of carbapenem-resistant Acinetobacter baumannii (CRAB) in hospitals, particularly within critical care units, has garnered substantial global concern. CRAB commonly arises from the degradation by various ß-lactamases. Objective: We aimed to assess OXA-type carbapenemases in clinical isolates of A. baumannii obtained from an Egyptian tertiary care facility. Patients and Methods: This study examined 25 distinct A. baumannii strains collected from various clinical samples of patients in intensive care unit. Bacterial identification was conducted utilizing both traditional methods and the Vitek2 system. Antibiotic resistance profiles were assessed according to the European Committee on Antimicrobial Susceptibility Testing standards using the Vitek2 Compact automated system. Additionally, multiplex real-time polymerase chain reaction was used to identify the presence of blaOXA23, blaOXA24, blaOXA51, and blaOXA58 carbapenemase genes. Colistin susceptibility was assessed utilizing the broth microdilution method. Results: Carbapenem resistance was identified in 100% of the studied isolates. The blaOXA51 gene was detected in all A. baumannii strains. The gene blaOXA23 was identified in 22 strains (88%), whereas blaOXA24 and blaOXA58 were present in 15 strains (60%). All isolates, except one, co-harbored two or more OXA encoding genes. Colistin resistance was detected in 4 of 25 strains (16%). Conclusion: Our findings demonstrate the widespread distribution of CRAB isolates that co-harbor multiple carbapenemase-encoding genes. Molecular epidemiological studies and the surveillance of antibiotic resistance profiles may aid in identifying and tracing the origins of resistant bacteria, thereby limiting their spread.

Improving Turnaround Times for Routine Antimicrobial Sensitivity Testing Following European Committee on Antimicrobial Susceptibility Testing Methodology in Patients with Bacteraemia.

Background/Objectives: Bacteraemia can be fatal without antibiotic intervention. Antibiotic Susceptibility Testing (AST) provides the necessary information for targeted antibiotic therapy; however, the traditional method using disc diffusion can take over two days from a positive blood culture. Inappropriate empiric therapy is associated with increased mortality and increased antibiotic resistance, highlighting the need for more rapid turnaround times for AST. By making changes to an established method, turnaround times can be reduced. Methods: Eighty-two patient positive blood culture samples were collected from January to April 2022, representing the range of common bacteria causing sepsis. This followed the normal methodology in the laboratory of inoculating agar from positive blood cultures in preparation for European Committee on Antimicrobial Susceptibility Testing (EUCAST) disc diffusion AST method. EUCAST methodology outlines that disc diffusion should be performed on isolates from an overnight culture of 16-24 h. This study looked at comparing disc diffusion results from cultures with 6 h of incubation to those with incubation times of 24 h, after organism identification by MALDI-ToF. Results from 6-h and 24-h cultures were compared by disc zone sizes and by interpreted susceptibility reading following EUCAST guidelines of sensitive, resistant, susceptible with increased exposure, or an area of technical uncertainty. Results: A total of 99.65% interpreted susceptibility readings matched across all organisms to all relevant antibiotics, with an average zone size difference of 1.08 mm between results from 6 h versus 24 h cultures. Conclusions: This method offers a non-automated way of using the traditional disc diffusion method, reducing turnaround times while still producing reliable and accurate results. This would mean validated ASTs can be set up in the same day as a blood culture flags positive rather than waiting for a longer culture. As this method is widely used within the laboratory already, it would mean that additional training is not required, as the process is the same, and only incubation time varies. This would positively impact patient outlook due to the shorter use of empiric therapy, and benefit antimicrobial stewardship (AMS).

GPT-4-based AI agents-the new expert system for detection of antimicrobial resistance mechanisms?

The study titled "GPT-4-based AI agents-the new expert system for detection of antimicrobial resistance mechanisms?" is critically important as it explores the integration of advanced artificial intelligence (AI) technologies, like generative pre-trained transformer (GPT)-4, into the field of laboratory medicine, specifically in the diagnostics of antimicrobial resistance (AMR). With the growing challenge of AMR, there is a pressing need for innovative solutions that can enhance diagnostic accuracy and efficiency. This research assesses the capability of AI to support the existing two-step confirmatory process recommended by the European Committee on Antimicrobial Susceptibility Testing for detecting beta-lactamases in Gram-negative bacteria. By potentially speeding up and improving the precision of initial screenings, AI could reduce the time to appropriate treatment interventions. Furthermore, this study is vital for validating the reliability and safety of AI tools in clinical settings, ensuring they meet stringent regulatory standards before they can be broadly implemented. This could herald a significant shift in how laboratory diagnostics are performed, ultimately leading to better patient outcomes.

Non-invasive Streptococcus pneumoniae infections are associated with different serotypes than invasive infections, Belgium, 2020 to 2023.

BackgroundDespite widely implemented pneumococcal vaccination programmes, Streptococcus pneumoniae remains a global risk for human health. Streptococcus pneumoniae can cause invasive (IPD) or non-invasive pneumococcal disease (NIPD). Surveillance is mainly focusing on IPD, assessing the full impact of pneumococcal vaccination programmes on pneumococcal disease is challenging.AimWe aimed to prospectively investigate serotype distribution and antimicrobial resistance (AMR) of S. pneumoniae isolates from patients with NIPD and compare with data on IPD isolates and with a 2007-2008 dataset on NIPD.MethodsBetween September 2020 and April 2023, we collected isolates and patient data from patients with NIPD from 23 clinical laboratories in Belgium. Capsular typing was performed by a validated Fourier-Transform Infrared spectroscopic method, and AMR was assessed with broth microdilution, using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) clinical breakpoints.ResultsWe received S. pneumoniae isolates from 1,008 patients with lower respiratory tract infections (n = 760), otitis media (n = 190) and sinusitis (n = 58). Serotype 3 was the most prevalent serotype among the NIPD isolates. Serotypes not included in the 20-valent pneumococcal conjugate vaccine (PCV20) were significantly more common among the NIPD than among the IPD isolates. Antimicrobial resistance levels were significantly higher among the NIPD isolates (n = 539; 2020-2022) compared with the IPD isolates (n = 2,344; 2021-2022). Resistance to several β-lactam antimicrobials had increased significantly compared with 15 years before.ConclusionsThe NIPD isolates were strongly associated with non-vaccine serotypes and with increased AMR levels. This underlines the importance of continued NIPD surveillance for informed policy making on vaccination programmes.

Antibiotic susceptibility testing using minimum inhibitory concentration (MIC) assays

Antimicrobial resistance is due to genetic changes that allow bacteria to evade antibiotic treatment. Antimicrobial susceptibility testing is critical for the detection of antibiotic-resistant strains, the selection of effective therapeutic strategies against bacterial infections, and the evaluation of the efficacy of novel antimicrobials. Among the variety of clinical microbiology methods used for antibiotic susceptibility testing, minimum inhibitory concentration (MIC) assays have become the gold standard in clinical practice. MIC assays determine the lowest concentration of an antimicrobial agent that is required to inhibit visible bacterial growth in vitro. Here, we outline MIC assay protocols, in strict accordance with European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines that aim to assess the susceptibility of non-fastidious organisms to antimicrobial agents. The protocols described in this methods paper are intended to aid the performance of reliable and informative MIC assays for research purposes that are in line with clinical microbiology practices.

Avian Pathogenic Escherichia coli Isolated in Poultry Farms in Bangladesh that Use Antibiotics Extensively.

Colibacillosis caused by avian pathogenic Escherichia coli (APEC) is causing economic losses to the global poultry industry. Increased prevalence of antibiotic resistance in APEC is the leading cause for increased indiscriminate use of various antimicrobial compounds in farms. The study aimed to investigate the presence of phenotypic and genotypic markers for antibiotic resistance, metals, and biocides in APEC from Bangladeshi poultry and details about the antimicrobials used in poultry farms. In total, 55 APEC were isolated from hearts or liver samples of 86 sick or dead chickens using culture on agar plate and biochemical testing. APEC isolates were tested for antibiotic susceptibility to 14 antimicrobial agents according to the European Committee on Antimicrobial Susceptibility Testing. A series of PCRs was performed to screen the presence of genes for quinolones, colistin, aminoglycosides, ESBL, metals, and biocides. Detailed information regarding antibiotic use was collected from farmers during clinical investigations. Resistance was found to 10 antibiotics and prevalence was as follows: ampicillin (86%), ciprofloxacin (86%), trimethoprim-sulphamethoxazole (73%), chloramphenicol (33%), mecillinam (13%), gentamicin (11%), cefoxitin (11%), cefotaxime (9%), tigecycline (2%), and nitrofurantoin (2%). The most common multiresistance phenotype was CIP-AMP-SXT, and 35% of isolates were multidrug resistant. Genotypic analysis confirmed the presence of quinolone resistance genes [qnrS1 and aac-(6')-lb-cr], silver-resistant genes (silE), and mercury-resistant genes (merA) but not others. In total, 88% farmers were using different antimicrobial compounds, and, of them, 56% were using antimicrobials without prescriptions from veterinarians. Ciprofloxacin was most extensively used followed by oxytetracycline. Critically important antibiotics like ciprofloxacin, colistin, and gentamicin are extensively used in the farms. This study confirmed the presence of antibiotics, metals, and biocide-resistant APEC in poultry farms in Bangladesh. Increased resistance to quinolones is a serious ongoing problem. The indiscriminate use of antibiotics in poultry farms is alarming and should be stopped.

In vitro activity assessment of cefiderocol against Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter spp., including β-lactam nonsusceptible molecularly characterized isolates, collected from 2020 to 2021 in the United States and European hospitals.

This study reports the activity of cefiderocol against Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter spp. isolates collected from the United States and Europe, including Israel and Turkey, from 2020 to 2021. Among Enterobacterales, 2.8% were carbapenem nonsusceptible (CNSE); cefiderocol inhibited 96.6%/85.1% [Clinical Laboratory Standards Institute (CLSI)/European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints] of these isolates. Imipenem-relebactam, meropenem-vaborbactam, and ceftazidime-avibactam displayed susceptibilities lower than cefiderocol against CNSE isolates (67.4-84.6% susceptible, CLSI). Cefiderocol was the only agent active against CNSE isolates carrying metallo-β-lactamase (MBL) carbapenemase or multiple carbapenemase genes (84.6%-92.3% susceptible, CLSI). Approximately 18% of carbapenem-susceptible Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis carried extended-spectrum-β-lactamases and/or plasmid-borne AmpC-encoding genes; cefiderocol inhibited 99.8%-100.0% (CLSI) of these genotypic groups. Multi-drug resistance (MDR) phenotypes were observed in 16.9% and 52.5% of P. aeruginosa and A. baumannii-calcoaceticus isolates, respectively. Carbapenemase genes were rare (4.9%) among cephalosporin and/or carbapenem nonsusceptible P. aeruginosa, compared to 87.6% carriage in A. baumannii-calcoaceticus, respectively. Against the MDR and carbapenemase-carrying P. aeruginosa and A. baumannii-calcoaceticus subsets, cefiderocol was active against 98.6%/98.7% and 97.1%/97.4% (CLSI), respectively. Only 69 isolates (0.3%) across all species groups were identified as cefiderocol nonsusceptible per CLSI criteria (>4 mg/L). Cefiderocol was the most active agent in vitro against Enterobacterales, P. aeruginosa, and Acinetobacter spp., with uniform activity against all phenotypic- and genotypic-resistant subsets. Coupled with the low incidence of nonsusceptibility observed across species groups, these results demonstrate cefiderocol as an important option for treating infections caused by pathogens with diverse mechanisms of resistance in US and European hospitals.IMPORTANCEThe worldwide spread of multi-drug-resistant Pseudomonas aeruginosa and carbapenem-resistant Enterobacterales and Acinetobacter spp. poses a serious challenge in healthcare settings as infections caused by these organisms are commonly refractory to many frontline therapeutic agents. Multiple global health organizations highlighted these pathogens as critical priorities for new antibiotic development, thus necessitating continued surveillance of the activities of currently available antimicrobial agents and circulating mechanisms of resistance. To meet this need, this study phenotypically and genotypically characterized priority Gram-negative pathogens collected from patients in US and European hospitals to examine the activity of cefiderocol and other currently available treatment options, including carbapenems and β-lactam-β-lactamase inhibitor combinations. The results presented here provide a detailed perspective to healthcare practitioners of cefiderocol's broad applicability, manifested in high activity and low nonsusceptibility rates, across phenotypic and genotypic organism groups relative to other agents and further support its use against the most intransigent infections.

Fosfomycin-Overcoming Problematic In Vitro Susceptibility Testing and Tricky Result Interpretation: Comparison of Three Fosfomycin Susceptibility Testing Methods.

Background: Fosfomycin (FOS) is an older antimicrobial agent newly rediscovered as a possible treatment for infections with limited therapeutic options (e.g., Gram-negative bacteria with difficult-to-treat resistance, DTR), especially in intravenous form. However, for correct usage of FOS, it is necessary to have a reliable susceptibility testing method suitable for routine practice and robust interpretation criteria. Results: The results were interpreted according to 2023 interpretation criteria provided by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). DTR Gram-negatives were more likely to be resistant to FOS (45% in Enterobacterales and 20% in P. aeruginosa) than non-DTR (10% and 6.7%, resp.). All isolates of S. aureus were susceptible to FOS. In Gram-negatives, all agreement values were unacceptable. Etest® performed better in the DTR cohort (categorical agreement, CA, 80%) than in the non-DTR cohort (CA 45.7%). There were no very major errors (VREs) observed in P. aeruginosa. S. aureus had surprisingly low essential agreement (EA) rates (53% for MRSA and 47% for MSSA) for Etest®, but categorical agreement was 100%. Methods: A total of 130 bacterial isolates were tested and compared using the disc diffusion method (DD) and gradient strip method (Etest®) with the reference method (agar dilution, AD). The spectrum of isolates tested was as follows: 40 Enterobacterales (20 DTR vs. 20 non-DTR), 30 Pseudomonas aeruginosa (15 DTR vs. 15 non-DTR), and 60 Staphylococcus aureus (30 methicillin-susceptible, MSSA, vs. 30 methicillin-resistant, MRSA). Conclusions: Neither one of the tested methods was identified as a suitable alternative to AD. It would be beneficial to define more interpretation criteria, at least in some instances.

Daily requests and complaints in Spanish prisons: Looking beyond legal regulation

AbstractThis article deals with the right to make requests and complaints (RCs) inside Spanish prisons. It discusses the European Prison Rules and the principles set forward by the European Committee for the Prevention of Torture and Inhuman or Degrading Treatment or Punishment (CPT), examining how the Spanish system respects these. It then introduces the RCs system in Spain. Through research carried out in four prisons, interviews with the directors and with 21 inmates and taking as an example one prison, we were able to discover that the number of requests in one year is 55,000. This result reveals the formalised character of Spanish prisons and raises the question as to how to respond to RCs inside prison. The last part of the article draws on interviews with three Penitentiary Judges and discusses their role in reviewing RCs. In our analysis of 626 of the final decisions we found that 98.6% were denied. Finally, the article provides some explanations for this result and questions whether judges can be said to provide an effective remedy in such cases.

Validation of EUCAST rapid antimicrobial susceptibility testing directly from positive blood cultures in a non-automated lab setting

ABSTRACT Introduction To speed up antimicrobial susceptibility testing (AST), the European Committee on Antimicrobial Susceptibility Testing (EUCAST) proposed rapid AST (RAST), a disk diffusion method to be read after 4, 6 and 8 hours of incubation. We investigated the feasibility of implementation of RAST in a non-automated lab setting. Materials & methods To this end, reference strains as well as a variety of clinical and resistant strains were used to spike sterile hemocultures (BioMérieux BACT/ALERT 3D® and Becton Dickinson BACTEC FX® systems), followed by RAST in comparison to classical long-incubation AST. Results & conclusion Our results with reference strains show that reading RAST after 4 hours is frequently too soon to obtain clinical results, and that Streptococcus pneumoniae reference strain did yield readable inhibition zones in RAST when harvested from BioMérieux BACT/ALERT 3D® bottle cultures. In a wider panel of strains, Gram positives RAST results were very similar to standard AST, while with Gram negative species errors were more frequently observed, limiting clinical implementation.

Honeycomb Molding - Forming Thermoplastic Sandwich Structures for Interior Applications

Trains play an important role in the sustainable mobility of the future. The changes in the market are reflected in the number of newly approved series by the German Federal Railway Authority (EBA); 226 new designs and redesigns of train cars were approved for use on German rails in 2019. Phenol-formaldehyde resins are used extensively specifically for railcar interiors. However, phenol-formaldehyde resins release carcinogenic vapors during and even after production if not completely sealed. As a result, European production standards and requirements have been increased, making the material less economical and increasingly difficult to enter the market. Further bans aimed at lower formaldehyde release limits are currently being considered by the European Union's Committee for Risk Assessment (RAC) and Committee for Socioeconomic Analysis (SEAC). In order to develop and design alternative materials, it is important to meet the requirements for stiffness, light weight, insulation and sustainability simultaneously. This research aim is therefore to develop a new class of material for interior components with improved stiffness, lower weight, lower CO2 footprint and no health hazards. The approach is to develop advanced composite sandwich panels that retain the ability to be molded into complex structures for interior applications. This is done through a thermoplastic honeycomb core structure that is manufactured directly from sheet material and fiber reinforced organo-sheets as the outer layers. The base matrix for all layers is kept uniform for better recyclability and consists of polycarbonate (PC) with sufficient flame retardant properties to provide an environmentally friendly alternative to existing phenolic resin and aramid epoxy honeycomb solutions while still complying with the security norms.