Background Hospital sink drains are increasingly recognized as important environmental reservoirs of multidrug-resistant Gram-negative bacteria and contributors to healthcare-associated infections. Conventional chemical disinfection strategies have limited and transient efficacy and contribute to environmental pollution through the release of chemical biocides. Probiotic-based approaches relying on competitive exclusion represent a promising alternative for the sustainable control of hospital microbial reservoirs. Material and methods In this study, we evaluated the inhibitory activity of two probiotic strains, Bacillus amyloliquefaciens NBS31007 and Bacillus velezensis S499, against nineteen strains of Gram-negative pathogens isolated from hospital sink drains. They belonged to the species Klebsiella pneumoniae , Enterobacter cloacae complex, Citrobacter freundii , and Pseudomonas aeruginosa , and produced extended-spectrum β-lactamases (ESBL) or carbapenemases. Reference strains of each species were used as controls. Bacillus strains were pre-cultured under oligotrophic conditions mimicking sink drain environments and subsequently challenged with low inocula of pathogenic strains. Pathogen growth was quantified after 24 hours of co-culture, and compared with growth in the absence of Bacillus . Results Both Bacillus strains inhibited the growth of all tested drain-related and reference strains, with reductions ranging from 1.0 to 6.1 log₁₀ CFU reduction ( P -values < 6.51 10 − 4 ) depending on the bacterial species and isolate. Overall, B. amyloliquefaciens NBS31007 and B. velezensis S499 displayed comparable inhibitory capacities, with only limited strain-dependent differences. Conclusions These findings provide proof of concept that probiotic Bacillus pre-cultures can limit the establishment of ESBL- and carbapenemase-producing Gram-negative pathogens in hospital sink drains. This biocontrol strategy may represent a complementary infection prevention approach aligned with antimicrobial resistance mitigation and One Health principles.

Antimicrobial resistance is a serious global health threat, yet the drivers of its spread among humans are not fully understood. Antibiotics can enter the human gastrointestinal tract through the food chain, leading to the presence of low concentrations in the gut microbiota. However, the role of such traces in promoting the implantation of drug-resistant pathogens in the gut microbiota has never been explored in a controlled experimental setting. Using an in vitro model of the human gut microbiota, we tested whether traces of 19 antibiotics used in both human and veterinary medicine, alone or in combination, lead to the enrichment of Gram-negative pathogens producing extended-spectrum β-lactamases or carbapenemases. Twenty-eight strains of Gram-negative pathogens epidemic in humans (10 Escherichia coli, 6 Klebsiella pneumoniae, 5 Enterobacter hormaechei, 4 Acinetobacter baumannii, 3 Pseudomonas aeruginosa) were tested. We found that antibiotics at levels similar to those measured in the feces of healthy individuals (fluoroquinolones, 1-100 μg L-1; trimethoprim, 100 μg L-1; a mixture of fifteen veterinary antibiotics, 10-20 μg L-1) enriched the human gut microbiota with those resistant pathogens. Overall, the present study indicates that dietary consumption of some antibiotics can result in concentrations in the human colon sufficiently high to favor the implantation of exogenous antibiotic-resistant pathogens. These findings highlight the need to reassess permissible antibiotic concentrations in food and critically evaluate agricultural practices contributing to the contamination of animal- and plant-based products.

BACKGROUND:The occurrence of foreign body (FB) infections in patients with Staphylococcus aureus bacteraemia (SAB) can lead to serious complications. We therefore sought to evaluate the host risk factors and bacterial determinants associated with FB infection following SAB. METHODS:We retrospectively included patients hospitalized with a SAB and carrying FB, over a four-year period. Factors associated with FB infection were assessed with multivariable logistic regression analysis. Bacterial determinants (lineage and virulence factors) were determined using whole genome sequencing. RESULTS:We included 104 patients of whom 40 patients (38.5%) with FB infection. Factors associated with FB infection were community-acquired SAB (OR = 3.8; 95% confidence interval, CI [1.36-10.59]; p = 0.011), Charlson's score >3 (OR = 0.39; 95% CI [0.16.-0.99]; p = 0.048 and unknown source of infection (OR = 3.54; 95% CI [1.34-9.33]; p = 0.011). No clonal complex or virulence gene was associated with a risk of FB infection in patients with SAB. CONCLUSION:FB infection is frequent in patients with SAB, particularly when the bacteremia is community-acquired. Our results suggest that bacterial characteristics (clonal complex or virulence factors) are not involved in the occurrence of FB infection in patients with SAB.

Legionnaires’ disease is a severe respiratory infection that causes increased mortality in hospitalized and immunocompromised patients. We report a nosocomial case in France linked to a water flosser. Our findings underscore the need for vigilance regarding such devices and highlight strategies for their safe management within healthcare settings.

OBJECTIVES:We aimed to assess the impact of COVID-19 on antibiotic consumption (AMC) and antimicrobial resistance (AMR) in the new epidemiological scenario from a cross-national perspective. METHODS:A quasi-experimental retrospective multicentre ecological study was conducted to explore the impact of COVID-19 on AMC and AMR using routinely generated retrospective time series data. This study included nine Healthcare University Hospitals from Europe and Israel on behalf QUantifying change in Antibiotic Resistance, ANTibiotic use, and INfection control during COVID-19 Epidemics project. Total effects were defined as the difference between the pre-COVID-19 period (ranging from January 2015 or January 2016 to February 2020) and during the COVID-19 pandemic period (March 2020 to July 2021 or December 2021). The outcomes were incidence density (ID) of carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Klebsiella pneumoniae, extended-spectrum beta-lactamase-producing Escherichia coli, vancomycin-resistant enterococci (VRE), methicillin-resistant Staphylococcus aureus, carbapenem-resistant Pseudomonas aeruginosa and Clostridioides difficile, as monthly isolates per 1000 patient days and the monthly AMC ranked according to the Access, Watch, and Reserve WHO classification system. RESULTS:We assessed 15.9 million total hospital bed days, 315 736 COVID-19 bed days, 52 557 monthly bacterial isolates, and 461 739 monthly antimicrobial defined daily doses. The COVID-19 pandemic had a significant impact on the consumption of overall hospital antibiotics combined in all centres except two. Prescriptions for piperacillin/tazobactam, glycopeptides, and ceftazidime/avibactam increased, whereas third-generation cephalosporins, macrolides, and fluoroquinolones returned to pre-pandemic levels after an initial surge, in all centres. A positive relationship between the pandemic intensity and VRE ID was observed in 6 of 9 (66%) centres followed by methicillin-resistant S. aureus-ID and carbapenem-resistant P. aeruginosa-ID 3 of 4 (44%) for both. A negative relationship was found for extended-spectrum beta-lactamase-producing E. coli ID. DISCUSSION:The COVID-19 pandemic was associated with higher usage of broad-spectrum antibiotics and higher incidence of multidrug-resistant bacteria, with great variability by countries. These results could support international action plans that embed AMR as a priority in the post-COVID-19 era.

In high-income countries, dogs and cats are often considered members of the family. Because of this proximity, it has been suggested that pets and humans might exchange bacterial species from their gut microbiota, with multidrug resistant bacteria being of particular concern. The aim of this study was to compare the genomes of third-generation cephalosporin-resistant (3GC-R) Escherichia coli responsible for human and pet infections in Europe.Whole-genome sequencing data from 3GC-R E. coli isolated from clinical samples of humans, dogs and cats, and published in eight European studies were re-analyzed using bioinformatics tools. The acquired genes responsible for 3GC-R were identified. The sequence type (ST) of all genomes were assessed by multilocus sequence typing. Alpha and beta diversities were measured within and between the two populations.We included genomes of 1327 3GC-R E. coli isolated from humans and animals with 109 (8.2 %) being responsible for infections in dogs and cat, and 1218 (91.8 %) responsible for human infections. Alpha diversity analysis suggested greater diversity within ST and 3GC-R genes in the animal population. Beta diversity analysis by principal coordinate analysis separated animal and human strains. ST131 was more abundant in human strains (43.4 %) than in animal strains (14.7 %) (p < 0.001). Six STs, including ST372, were identified almost exclusively in 3GC-R E. coli from animal origin. The blaCTX-M-15 gene was more frequent in humans (49.24 %) than in companion animals (17.9 %) (p < 0.001). The resistance genes blaCMY-2 (30.8 %) and blaCTX-M-1 (15.4 %) were more frequent in E. coli isolated from pets (p < 0.001).We found that populations of 3GC-R E. coli responsible for human and pet infections in Europe do not overlap. Although it cannot rule out occasional transmission of bacteria between pets and humans within a household, it suggests that dogs and cats are not a major source of human infection with this antibiotic-resistant pathogen.

INTRODUCTION:Urinary tract infection (UTI) in men, although less common than in women, present specific diagnostic and therapeutic challenges. This study aims to evaluate the prescribing practices of general practitioners (GPs) for male UTI in France, focusing on adherence to guidelines. MATERIALS AND METHODS:We used an anonymous reimbursement database of antibiotics prescribed 15 days around an urine culture between September 2019 and August 2022 in a French region. Antibiotic prescriptions for male UTI were analysed according to adherence to national guidelines. Cluster analysis was used to identify different GP prescribing profiles. Prescription duration was assessed according to the number of antibiotic boxes delivered in the community pharmacy. RESULTS:We included 7,816 urine culture prescriptions from 940 GPs for 6,457 male patients. We estimated compliance with French recommendations to be 55.7% for empirical treatment and 68.1% for documented treatment. GPs were divided into three clusters with different adherence to recommendations of 22%, 44% and 77%. Treatment duration for fluoroquinolones and cotrimoxazole was heterogeneous between GPs, but mainly too short. CONCLUSIONS:Our results suggest that our method could identify GPs who do not prescribe in accordance with recommendations and enable health insurance systems to target educational interventions to improve antibiotic prescribing practices.

Controlling the spread of bacterial infectious diseases is a major public health issue, particularly in view of the pandemic of bacterial resistance to antibiotics. In this context, the detection and identification of pathogenic bacteria is a prerequisite for the implementation of control measures. Current reference methods are mainly based on culture methods, which generate a delay in obtaining a result and requires equipment. Consequently, focusing on the detection of the whole bacterium represents a very attractive alternative, since no culture is required. Several techniques have already been deployed to identify whole-cell bacteria. In recent decades, growing interest in nucleic acid aptamers has emerged as a viable alternative to antibodies as recognition elements, offering preferable stability, cost-efficiency, good specificity and affinity. This review explores current alternative methods for the detection of whole-cell bacteria, with particular emphasis on aptamer-based assays. These assays have shown promising results in various transduction mechanisms, including optical, electrochemical, and mechanical approaches, enhancing their versatility in different diagnostic platforms. The integration of aptamers in these detection methods offers rapid, sensitive, versatile and portable solutions for pathogen identification, positioning them as valuable tools in the fight against bacterial infections.

To evaluate the utility of automated urine flow cytometry in the diagnosis of urinary tract infection (UTI) in older patients without waiting for urine culture results. This prospective study included patients aged ≥ 65 years admitted to the emergency department of Besançon University Hospital over a six-month period. Clinical and biological data were collected and UTI diagnosis was based on strict clinical and biological criteria. Urine analysis was performed using the UF-4000 (Sysmex). Parameters or thresholds were defined based on an AUC > 0.8 and a clinically relevant negative likelihood ratio (LR-) < 0.1. Of 456 patients, 69 (15.1

Background: Surveillance of antimicrobial resistance (AMR) in healthcare settings is essential for effective management of infections, optimization of antibiotic treatments and early detection of hospital outbreaks. Traditional surveillance for patients, based on routine clinical antimicrobial susceptibility data and multidrug-resistant organisms (MDRO) screening, has limitations. Wastewater-based surveillance (WBS) has emerged as an alternative tool for benchmarking health markers, as used in wastewater monitoring of virus around the COVID-19 pandemic. WBS can potentially serve as a cost-effective and efficient tool for monitoring resistant pathogens and genes in hospital settings. Objectives: Here we review the role of WBS for monitoring AMR in hospitals, summarizing current evidence and challenges. Practical recommendations are also provided on what must be considered when implementing AMR WBS programs in hospitals, including system setup, selection of surveillance targets, wastewater collection methods, microbiological processing, and interpretation of WBS-generated data. Results and implications: WBS could serve as a promising tool for monitoring AMR in hospitals, with the potential to enhance infection control efforts. However, the optimal and validated approach for this purpose has yet to be established. There is significant variability in the methods used for sampling and processing of wastewater, and ideal sampling techniques and microbiological methods for tracking AMR need to be determined. Key knowledge gaps remain regarding the qualitative and quantitative link between AMR detected in hospital wastewater, and hospitalized patients. This link can be influenced by various factors, including presence of antibiotic residues, or biofilm formation in sanitary or effluent pipes and external environmental factors such as rainwater mixing. New research is needed to refine WBS methods for AMR, to better understand the link between wastewater findings and prevalence of AMR in hospitalized patients and to validate WBS effectiveness as a health surveillance strategy, or instrument for benchmarking of AMR prevalence in hospital populations.

OBJECTIVE:To determine the most sensitive and feasible method for surveillance of the hospital aquatic environment for detection of carbapenemase-producing Enterobacterales (CPE) and extended-spectrum β-lactamase-producing Enterobacterales (ESBLE). METHODS:This study focused on hospital sink P-traps and toilets, comparing the yield of liquid collection with swabbing for sampling. Additionally, sample processing by direct culturing on selective media was compared with culturing following overnight enrichment. Bacterial species were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry, and the production of ESBLE or CPE was confirmed using phenotypic or immunochromogenic methods. RESULTS:In total, 32 sink P-traps and 20 toilets were sampled in two wards of a university hospital. The different sampling techniques (liquid collection and filtering, swabbing) and processing methods (without or with cefotaxime, temocillin/imipenem enrichment) were applied, separately for ESBLE and CPE detection, for all sinks and toilets, resulting in 520 samples. Of the 32 sink P-traps sampled, 21 (65.6%) were positive for ESBLE and eight were positive for CPE (25.0%) with at least one of the sample methods. Of the 20 toilets sampled, 10 (50.0%) were ESBLE positive and nine (45.0%) were CPE positive. For sink P-traps, swabbing and liquid collection showed comparable results. Although enrichment only led to a marginal increase in detection sensitivity, it facilitated the subsequent microbiological analysis. For toilets, water collection was more sensitive than rim swabbing. CONCLUSION:These results suggest that swabbing of sink P-traps and water collection from toilets, combined with enrichment, are the most suitable methods for the detection of multi-drug-resistant pathogens in the hospital aquatic environment.

Background: Staphylococcus haemolyticus is a clinically relevant coagulase-negative staphylococcus frequently responsible for hospital-acquired infections, especially in premature newborns. Aim: To describe an outbreak of multidrug-resistant S. haemolyticus in a neonatal department. Methods: The outbreak was investigated using classical methods, including screening of the patients, genotyping of the isolates and environmental survey. Numerous infection control measures were implemented. Findings: In 2022 and 2023, a clonal outbreak of multidrug-resistant S. haemolyticus ST29 (40 infections and 71 carriages) occurred in the neonatology department of a University Hospital in France. The infection control measures implemented only partially controlled the outbreak. Although our investigation did not clearly identify the source and mode of transmission, a reservoir constituted by patients and transmission by healthcare workers are the most likely. This episode occurred in a context of countrywide outbreaks of S. haemolyticus ST29 in several French neonatology departments. Conclusion: This prolonged outbreak of S. haemolyticus ST29 accounted for the increase in the incidence of S. haemolyticus-related infections in French neonatology departments. Implementation of proactive measures is crucial to limit the spread of such pathogens in neonatal ICUs. (c) 2024 The Healthcare Infection Society. Published by Elsevier Ltd. All rights are reserved, including those for text and data mining, AI training, and similar technologies.

Karst aquifers are a significant source of drinking water and highly vulnerable to pollution and microbial contamination. Microbiological regulations for the quality of drinking water mostly focus on bacterial levels and lack guidance concerning fungal contamination. Moreover, there is no standardised microbial analysis methodology for identifying fungi in water. Our main objective was to establish the most effective culture and identification methodology to examine yeast diversity in karst waters. We assessed the comparative efficacy of four culture media (CHROMagar Candida, dichloran glycerol 18% [DG18], dichloran rose Bengal chloramphenicol [DRBC], and SYMPHONY agar) for yeast isolation from karst water samples. Furthermore, we investigated the comprehensiveness of databases used in MALDI-TOF mass spectrometry (MALDI-TOF MS) for identifying environmental yeast species. In total, we analysed 162 water samples, allowing the identification of 2479 yeast isolates. We demonstrate that a combination of four culture media, each with distinct specifications, more efficiently covers a wide range of yeast species in karst water than a combination of only two or three. Supplementation of a MALDI-TOF MS database is also critical for analysing environmental microbial samples and improved the identification of yeast biodiversity. This study is an initial step towards standardising the analysis of fungal biodiversity in karst waters, enabling a better understanding of the significance of this environmental reservoir in relation to public health.

BACKGROUND:Staphylococcus haemolyticus is a clinically relevant coagulase-negative staphylococcus frequently responsible for hospital-acquired infections, especially in premature newborns. AIM:To describe an outbreak of multidrug-resistant S. haemolyticus in a neonatal department. METHODS:The outbreak was investigated using classical methods, including screening of the patients, genotyping of the isolates and environmental survey. Numerous infection control measures were implemented. FINDINGS:In 2022 and 2023, a clonal outbreak of multidrug-resistant S. haemolyticus ST29 (40 infections and 71 carriages) occurred in the neonatology department of a University Hospital in France. The infection control measures implemented only partially controlled the outbreak. Although our investigation did not clearly identify the source and mode of transmission, a reservoir constituted by patients and transmission by healthcare workers are the most likely. This episode occurred in a context of countrywide outbreaks of S. haemolyticus ST29 in several French neonatology departments. CONCLUSION:This prolonged outbreak of S. haemolyticus ST29 accounted for the increase in the incidence of S. haemolyticus-related infections in French neonatology departments. Implementation of proactive measures is crucial to limit the spread of such pathogens in neonatal ICUs.

Pyrolysis-gas chromatography-mass spectrometry (pyr-GC-MS) is emerging as a promising alternative for the detection and quantification of microplastic pollution. For a robust quantification it is essential to improve our understanding of interferences in the pyrolysis of microplastics. Here we investigate the effects of different soil matrices, mainly differing by their organic carbon content (Corg, 1.0-13.6%), and of the polymer molecular weight (Mw) on the pyr-GC-MS analysis of polystyrene (PS) microplastics. In addition, we evaluated the effectiveness of adding poly(4-fluorostyrene) (PSF) as internal standard to circumvent the matrix effects. The three main markers of PS pyrolysis, i.e., styrene, styrene-dimer and styrene-trimer, were monitored. The ratio between the dimer and the trimer significantly varied between the matrices and tended to decrease with the increasing of the Corg in the soil, mainly due to an increased trimer formation. A strong matrix effect affected the slope of the calibration curves by 2 to 8-fold and was correlated with the Corg in the soils. This effect was mitigated when the areas of the markers were normalized by the area of the corresponding marker of PSF. PS of low Mw (Mw 35,000) presented a reduced formation of the three markers compared to PS of high Mw (Mw 400,000), and styrene-dimer was proportionally less formed than the other two markers. Differences in the slopes of calibration curves depended on the marker chosen, highlighting the relevance of selecting the pyrolysis marker in the quantification of microplastics using pyr-GC-MS.

Pseudomonas aeruginosa is a major opportunistic pathogen responsible for severe infections in immunocompromised patients. The contamination of drinking water networks (DWNs) with this pathogen is underestimated, as it is mostly in the state of persister cells undetected by the recommended monitoring technique. We collected water samples from eight cities distant from each other and searched for P. aeruginosa using a culture-based method that resuscitates persister cells. The genomes of isolates were sequenced. Five DWNs of the eight tested (62.5%) were contaminated with P. aeruginosa, of which four were contaminated with high-risk clones (ST308, ST395). Surprisingly, the ST308 isolates retrieved from the four independent and distant DWNs were clonal. Most P. aeruginosa isolates shared a genomic island conferring tolerance to copper-ions. The population structure of the collection may result from both a common source of contamination by plumbing supplies and the selection of clones sharing genetic elements that presumably aided their propagation in DWNs.

OBJECTIVE:The objective of this study was to evaluate the ability of two macroscopic quality indicators to guide antibiotic stewardship strategies. METHODS:Setting: Bourgogne-Franche-Comté region (eight departments). Antibiotic reimbursement data, expressed in defined daily doses, were used to calculate two macroscopic quality indicators from 2015 to 2020. Urinary tract infection indicator: (fosfomycin + nitrofurantoin + pivmecillinam) / (ciprofloxacin + ofloxacin + norfloxacin) calculated in women aged 15 to 64. Indicator for ear, nose, throat and respiratory tract infections: (amoxicillin/ (amoxicillin-clavulanate + oral 3GC) calculated in children under 15 years of age. RESULTS:In 2020, global antibiotic consumption varied from 22.2 to 26.6 defined daily doses per 1000 inhabitants per day, depending on the department. The macroscopic quality indicators were lowest in the departments of Nièvre and Yonne. The scores for the urinary tract infection indicator were 0.55 and 0.74, respectively, while those for the ear, nose and throat and respiratory tract infection indicators were 1.85 and 2.12, respectively. Both scores continuously increased in all departments of the region during the survey, except in 2020 for the urinary tract infection indicator, due to a nitrofurantoin shortage. CONCLUSION:Even in departments with low antibiotic consumption, our macroscopic quality indicators help to identify antibiotic stewardship targets for the most common community-based infections,. They are easy to calculate from quantitative data without requiring any information on diagnosis. Complementarily to other indicators, they provide messages specifically addressed to general practitioners regarding population, molecules and diagnostic elements.

Objective: The aim of this study was to evaluate the ability of two macroscopic qualitative indicators (MQIs) to drive antibiotic stewardship strategies.Methods: Use of French regional antibiotic reimbursement data, expressed in defined daily doses, to calculate the two MQIs from 2015 to 2020. Urinary Tract Infections (UTI) indicator: (fosfomycin + nitrofurantoin + pivmecillinam) / (ciprofloxacin + ofloxacin + norfloxacin) calculated among women from 15 to 64 years old. Ears Nose Throat and Respiratory Tract Infection (ENT-RTI) indicator: (amoxicillin/ (amoxicillin clavulanate + oral 3GC) calculated among children under 15 years old.Results: In 2020, MQIs with low value, indicating nonoptimal prescription practices, was observed in Yonne department, whereas its global antibiotic consumption was among the lowest.Conclusion: Our MQIs contribute to identify targets of antibiotic stewardship for the most frequent community infections. They are easy to calculate from quantitative data without any information about the diagnosis.