Infection Control & Hospital Epidemiology
◐ Cambridge University Press (CUP)
Preprints posted in the last 30 days, ranked by how well they match Infection Control & Hospital Epidemiology's content profile, based on 17 papers previously published here. The average preprint has a 0.02% match score for this journal, so anything above that is already an above-average fit.
Shinto, H.; Chowell, G.; Takayama, Y.; Ohki, Y.; Saito, K.; Mizumoto, K.
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BackgroundIn long-term care facilities (LTCFs), close-contact identification often relies on staff recall and monitoring records because residents may be unable to self-report reliably. How these different record-generation processes relate to proximity-based sensor measurements in routine LTCF workflow remain unclear, and how such differences may influence contact-based decision-making in outbreak response is not well understood. MethodsWe conducted a five-day observational study in a Japanese LTCF using ultra-wideband (UWB) indoor positioning. Twenty-seven participants wore UWB tags, including 16 residents and 11 staff members; 10 staff members completed questionnaires. We compared UWB-derived proximity with questionnaire-derived contacts from staff self-report and monitoring-based proxy records, and assessed directional discrepancies under multiple distance-time thresholds. ResultsQuestionnaire-based records and UWB-derived proximity showed different patterns of discrepancy across contact types. Within this facility, resident-related monitoring-based proxy records showed relatively small directional discrepancies, whereas staff self-reports tended to identify additional resident-staff contacts under the baseline threshold ([≤]1.0 m for [≥]15 min). Several alternative thresholds were associated with discrepancies closer to zero than the baseline, although the apparent ranking varied by summary metric. ConclusionsIn this single-facility observational study, different contact-list generation processes were associated with different patterns of discrepancy relative to a proximity-based operational measure. These findings support interpretation in terms of workflow-specific contact-list generation rather than a single universally optimal threshold and may help inform facility-level review of contact identification practices in LTCFs. These findings support aligning contact identification strategies with facility-specific workflows to improve the feasibility and effectiveness of IPC practices in LTCFs.
Sheth, E.; Case, L.; Shaw, F.; Dwyer, N.; Poland, J.; Wan, Y.; Larru, B.
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Background Pseudomonas aeruginosa is a major cause of healthcare-associated infections in paediatric settings, where its persistence in moist environments such as hospital water and wastewater systems poses a particular risk to neonates and immunocompromised children. Aim The aim of this study was to showcase the long-term survival and transmission of P. aeruginosa in a large tertiary children's hospital in England which is crucial to develop strategies for water-safe care. Methods Environmental P. aeruginosa isolates were collected from taps, sinks, showers, and baths in augmented care areas of a 330-bed tertiary children's hospital built to NHS water-safety standards. Clinical isolates were classified as invasive (blood, cerebrospinal fluid, and bronchoalveolar lavage) or non-invasive (respiratory, urine, ear, abdominal, and rectal surveillance). Variable number tandem repeat (VNTR) profiles and metadata were extracted from PDF reports, de-identified, deduplicated, and curated using Python and R. Findings This retrospective study analysed nine-locus VNTR profiles of 457 P. aeruginosa isolates submitted to the UK Health Security Agency from a large tertiary children's hospital, identifying 56 isolate clusters (each with [≥]2 isolates), of which 19 (34%) contained at least one invasive isolate. The most persistent cluster (Cluster 1, n=20) spanned from July 2016 to September 2024, containing environmental and clinical (invasive and non-invasive) isolates. Conclusion These findings demonstrate long-term persistence of certain genotypes and temporal overlap between environmental and clinical isolates, highlighting the difficulty in detecting and eradicating P. aeruginosa in hospital water and wastewater systems and reinforcing the need for continuous rigorous water system controls.
Laskaris, Z.; Baron, S.; Markowitz, S. B.
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ObjectivesRising temperatures are a major climate-related hazard for U.S. workers, increasing heat-related illness and a broad range of occupational injuries through indirect pathways often overlooked in economic evaluations. We examined the association between temperature and occupational injury and illness and quantified heat-attributable injuries (including illnesses) and costs in New York State. MethodsWe conducted a time-stratified case-crossover study of 591,257 workers compensation (WC) claims during the warm season (2016-2024). Daily maximum temperature was linked to injury date and county and modeled using natural cubic splines, with effect modification by industry and worker characteristics. ResultsInjury risk increased with temperature, becoming statistically significant at approximately 78{degrees}F. Relative to 65{degrees}F, injury odds increased to 1.06 (95% CI: 1.01-1.10) at 80{degrees}F, 1.12 (1.07-1.18) at 90{degrees}F, and 1.17 (1.11-1.23) at 95{degrees}F. Overall, 5.0% of claims (2,322 annually) were attributable to heat. At temperatures [≥]80{degrees}F, an estimated 1,729 excess injuries occurred annually, generating approximately $46 million in WC costs. An estimated $3.2 million to $36.1 million in medical expenditures were associated with incomplete claims, likely borne outside the WC system. ConclusionsThese findings demonstrate substantial economic costs not fully captured within WC and support workplace heat protections as a cost-containment strategy that can reduce health care spending and strengthen workforce resilience.
Mills, E. A.; Bingham, R.; Nijman, R. G.; Sriskandan, S.
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BackgroundAn upsurge in Streptococcus pyogenes infections 2022-2023 highlighted potential benefits of point-of-care tests (POCT) to support clinical pathways, prevent outbreaks, and optimise antibiotic use. ObjectivesWe conducted a pilot research study in a west London paediatric emergency department (ED) to determine whether a molecular POCT had potential to alter management in children who were also having a conventional throat swab taken for culture. MethodsChildren <16 years presenting to ED who had a throat swab requested by a clinician were invited to have a second swab taken for research purposes only. Clinical management was unaffected by the research swab result, which was processed using a molecular POCT that was not approved for use in the host NHS Trust. ResultsPrevalence of streptococcal infection was low during the study (May 2023-June 2025); swab positivity in symptomatic children was 12.8% (6/47). Overall, 38/49 (77.6%) participants who had throat swabs received antibiotics. Of those children recommended to receive antibiotics, 29/38 (76.3%) had a negative POCT. Mean time to reporting of positive throat swab culture results was 3.67 days (range 3-5 days) leading to occasional delay in treatment, although POCT identified positive results within minutes. ConclusionAntibiotic use was frequent and could be avoided or stopped by use of a rule out POCT in over three-quarters of children in the ED, if suspicion of S. pyogenes is the main driver for prescribing. POCT were easy to process and produced immediate results compared with culture, in theory enabling timely decision-making and avoiding treatment delay.
Chhabra, S.; Nair, S.; Bramley, A.; Chee, J. Y.; Vignesvaran, K.; See, D. R. E.; Sun, L. J.; Ching, A. H.; Li,, A. Y.; Kayastha, G.; Chetchotisakd, P.; Cooper, B. S.; Charani, E.; Mo, Y.
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Background Antibiotic use is prevalent in hospitals, driving the emergence of drug-resistant pathogens. We investigated the contextual influences on antibiotic prescribing behaviour across hospitals in high, middle, and low-income countries in Asia with an aim to provide actionable insights to improve prescribing behaviour. Methods We conducted a large qualitative study across ten institutions in Singapore, Nepal, and Thailand. Semi-structured interviews and ethnographic observations involving physicians, nurses, pharmacists, and management staff were conducted. Data were analysed thematically using QSR NVivo 14. Findings A total of 194 interviews were conducted amongst physicians (54{middle dot}1%), nurses (19{middle dot}6%), pharmacists (12{middle dot}4%), and management staff (13{middle dot}9%). Structural factors such as limited microbiology laboratory capabilities, concerns about antibiotic quality, weak infection prevention and control policies, and the lack of relevant, updated guidelines were prominent drivers for prolonged and broad-spectrum antibiotics prescriptions. Where these system supports were in place, prescribing decisions were less defensive and more targeted, although prescriber responsibility and concerns about immediate patient deterioration continued to influence practice. Across settings, clinicians tended to prioritise short-term perceived benefits of antibiotic treatment over the longer-term risks of antimicrobial resistance.
Koudoum, P. L.; Ateudjieu, W. D.; Nana, A.; Guemkam, G. W.; Nditemeloung, G.; Abena, J. V.; Rene, E.; Vigny, N. N.; Joseph Magloire, T.; Mbossi, A. D.; Kamgno, J.; Kamga, H. G.
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Antimicrobial resistance (AMR) is classified by the World Health Organization (WHO) as one of humanity's ten global public health threats. This review aimed to estimate the prevalence, temporal trends and regional distribution of AMR in WHO priority bacteria across human, animal and environmental sources in Cameroon. This review was conducted following PRISMA 2020 guidelines, with the protocol registered in PROSPERO. A systematic literature search was conducted in Google Scholar, PubMed, African Journals Online, Hinari, and Africa indexus Medicus. Random effects models were used to estimate pooled prevalence and 95% confidence intervals (CIs), with subgroup analyses by bacterial source, region, and sampling period. Of 1566 articles screened, 115 met the inclusion criteria. The reported data encompassed 16 bacteria-antibiotic combinations in 16,948 isolates. Globally, third-generation cephalosporin (3GC) resistance in E. coli was the most prevalent (49.0%, 95% CI: 39.0-60.0%, I2=97.7%), reaching 77.0% (95% CI: 46.0-98.0%, I2=95.6%) in environmental isolates. The pooled prevalence of ESBL production in all included Enterobacterales was 37.0% (95% CI: 30.0-45.0%). Most of the highest resistance rates were observed in the Littoral region. The resistance rates between 2016 and 2025 were significantly higher than those from 2000 to 2015. These increases were more marked in fluoroquinolone-resistant Salmonella spp (1.0% to 48.0%, I2=97.3%, p<0.001), carbapenem-resistant E. coli (0% to 15%, I2=93.5%, p<0.001), and 3GC-resistant E. coli (34.0% to 64.0%, I2=97.6%, p=0.003). Antimicrobial resistance in WHO priority bacteria in Cameroon is high, unevenly distributed across regions and significantly increasing over time. These results underscore the crucial need for strengthened AMR surveillance to curb the growing threat of AMR in Cameroon.
Bajwa, H. U. R.; Bhowmick, S.; Varga, C.
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Introduction Nontyphoidal Salmonella enterica (NTS) is a major zoonotic enteric pathogen. Animal contact-related NTS outbreaks have increased in the United States of America (U.S.) over the last decade. Geospatial analysis can identify locations with elevated risk of NTS outbreaks where public health authorities can focus their NTS prevention and intervention efforts. Methods We analyzed NTS outbreak data reported from individual states to the Centers for Disease Control via the National Outbreak Reporting System between 2009 and 2022 across the continental contiguous U.S. A geospatial analytical framework that included disease mapping, spatial interpolation, and global and local clustering methods was applied to identify regions with high NTS outbreak rates. Results A total of 104 NTS single-state outbreaks were reported to the National Outbreak Reporting System (NORS) during the study period. The mean annual incidence rate was 0.02 NTS outbreaks per million person-years. The primary animal contact categories associated with these outbreaks were mammals (cattle, pigs, sheep, and horses), birds (backyard chickens, ducklings, and turkeys), and reptiles (turtles and lizards). Exposure settings included farms, fairgrounds, agricultural feed stores, veterinary clinics, dairy/agricultural settings, and residential settings. The local cluster detection methods consistently identified areas with significantly high NTS animal contact-related outbreak rates in the Mountain West, Midwest, and Northeast of the US. Conclusion NTS animal contact-related single-state outbreaks revealed distinct spatial clustering across the United States, with potentially higher risks in the Mountain West, Midwest, and Northeast. Diversity of animal-contact sources and exposure settings depicted complex transmission dynamics of NTS. Focused prevention and control programs in these areas are needed to mitigate the burden of NTS outbreaks.
Yu, J.; Tillema, S.; Akel, M.; Aron, A.; Espinosa, E.; Fisher, S. A.; Branche, T. N.; Mithal, L. B.; Hartmann, E. M.
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Benzalkonium chloride (BAC) is widely used as a disinfectant in cleaning products and is frequently detected in indoor dust. In this study, we assessed dust samples, along with information on cleaning product use, from 24 pregnant participants. Dust samples were analyzed for BAC concentration and microbial tolerance. Different chain lengths of BAC (C12, C14, and C16) were quantified using LC-MS/MS, and bacterial isolates were tested for BAC tolerance using minimum inhibitory concentration (MIC) assays. BAC was ubiquitously detected, with C12 and C14 being dominant. Higher BAC concentrations were associated with reported disinfectant use and increased microbial tolerance. These findings suggest that indoor antimicrobial use may promote microbial resistance, highlighting potential exposure risks in indoor environments and the need for further investigation into health and ecological impacts.
Rodgus, J.; Fraser-Krauss, O.; Ravindra, Y.; Getino, M.; Myall, A.; Yoon, C. H.; Upadhya, A.; Peach, R.; Mookerjee, S.; Holmes, A.; Jauneikaite, E.; Barahona, M.; Davies, F.
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Infections caused by carbapenem-producing Enterobacterales (CPEs) are a persistent and growing threat in healthcare settings. Yet, current infection prevention and control (IPC) surveillance methods, which largely rely on the spatial and temporal proximity of patients, often misattribute or miss infection transmission events. Here, we develop and retrospectively evaluate an integrated methodology that combines analyses of ward-level patient movement data and whole-genome sequencing (WGS) data analyses, providing measures of bacterial and plasmid similarity. Specifically, we evaluate this methodology across two datasets: a CPE outbreak of diverse carbapenem types (103 genomes, January 2021 to March 2021) and an Imipenem-Hydrolysing beta-lactamase-positive CPE outbreak (82 genomes, June 2016 to October 2019), using standard clinical criteria and conservative genomic thresholds to quantify how often current IPC surveillance methods correctly identify genomically confirmed transmission events. Findings show that, across 3,423 patient contact-genome pairs, current IPC surveillance methods detected only 20.5% of genomically confirmed transmission events whilst maintaining 98.5% specificity, with missed events arising from temporal, spatial, and cross-species, mechanistic blindspots. In contrast, WGS-enabled IPC surveillance methods provided a 25 to 47-day earlier detection window and, in a linked economic evaluation, delivered annualised savings of up to GBP 3.6 million, as well as a return on investment exceeding 2-fold in 7 of 8 cost scenarios. By operationalising high-throughput WGS data analysis with clinically relevant patient movement data, we evidence that it may be possible to disrupt and thereby mitigate the effects of AMR-driven CPE outbreaks, supporting investigations into the adoption of WGS-enabled IPC surveillance as a standard-of-care tool.
Watanabe, N.; Watari, T.; Otsuka, Y.; Matsumiya, T.
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Background Five-biomarker-defined hypervirulent Klebsiella pneumoniae (hvKp) causes invasive infections, but its burden in bloodstream infections versus classical K. pneumoniae (cKp) is unclear. Methods This retrospective cohort study at a tertiary hospital in Japan included K. pneumoniae bloodstream infection episodes from January 2022-December 2024. hvKp was defined by the presence of all 5 genotypic biomarkers (rmpA, rmpA2, iucA, iroB, and peg-344). The primary outcome was abscess complications, and secondary outcomes were length of stay and antibiotic duration. Whole-genome sequencing was performed for 164 isolates. Results Among the 207 episodes, 28 (14%) were of hvKp. Abscess complication occurred in 17 (61%) hvKp versus 23 (13%) cKp episodes (adjusted odds ratio 10.7; 95% CI, 4.36-26.2). Median length of stay in hvKp versus cKp was 28 versus 14 days (adjusted ratio 1.60; 95% CI, 1.18-2.16) and median antibiotic duration was 43 versus 14 days (adjusted ratio 2.13; 95% CI, 1.64-2.77). These associations were attenuated after adjusting for abscess-related complications. No significant difference in 30-day mortality was observed, although the study was underpowered. Multidrug resistance was less frequent in hvKp strains than in cKp strains (11% vs. 30%; P = .040). Among the sequenced hvKp episodes, abscess rates varied across lineages, from 9 of 10 in ST23 to 1 of 4 in ST412. Conclusions Five biomarker-defined hvKp strains delineated a bloodstream infection subgroup with frequent abscess complications and prolonged care. hvKp and cKp present distinct clinical challenges; diagnostic tools distinguishing these subgroups may aid abscess evaluation and source control.
Mogeni, P.; Ochieng, J. B.; Kariuki, K.; Rwigi, D.; Atlas, H. E.; Tickell, K. D.; Aluoch, L. R.; Sonye, C.; Apondi, E.; Ambila, L.; Diakhate, M. M.; Singa, B. O.; Liu, J.; Platts-Mills, J. A.; Saidi, Q.; Denno, D. M.; Fang, F. C.; Walson, J. L.; Houpt, E. R.; Pavlinac, P. B.
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BackgroundThe Toto Bora trial tested whether a course of azithromycin reduced rates of re-hospitalization or death in the 6 months following hospitalization among Kenyan children. We hypothesized that azithromycin would reduce enteric bacteria and increase carriage of macrolide resistance in the subsequent 3 months. MethodsKenyan children (1-59 months) hospitalized and subsequently discharged for non-traumatic conditions provided fecal samples before and 3 months after randomization to a 5-day course of azithromycin or placebo. Quantitative PCR identified enteropathogens and AMR-conferring genes in fecal samples. Generalized estimating equations assessed the impact of the randomization arm on pathogen and resistance gene detection, accounting for baseline presence and site. ResultsAmong 1,393 baseline stools, 12.4% had at least one bacterial enteropathogen, 94.7% had at least one macrolide-resistance gene, and 92.6% had at least one beta-lactamase-resistance gene identified. At month 3, children randomized to azithromycin had a 6.1% higher likelihood of carrying a macrolide resistance gene compared to placebo (adjusted prevalence ratio [aPR], 1.06; 95% CI, 1.04-1.08; P<0.001). Specifically, azithromycin randomization was associated with a higher relative prevalence of erm(B) (aPR, 1.09 [95% CI, 1.04-1.15]; P=0.001), erm(C) (aPR, 1.23 [95% CI, 1.14-1.31]; P<0.001), msr(A) (aPR, 1.14 [95% CI, 1.04-1.25]; P=0.007), and msr(D) (aPR, 1.07 [95% CI, 1.03-1.11]; P=0.001). There was no difference in overall bacterial pathogen prevalence (18.9% vs 17.3%) between randomization arms, but a slightly lower proportion of children had Shigella after randomization in the azithromycin arm (3% vs. 5%, aPR, 0.79 [95% CI, 0.62, 1.01]; P=0.063). InterpretationAzithromycin at hospital discharge was associated with higher carriage of macrolide-resistance-conferring genes in the post-discharge period compared with placebo, without significant declines in enteric pathogen carriage other than modest changes to Shigella. The potential benefits and risks of empiric azithromycin need to be considered, as children are increasingly exposed to this broad-spectrum antibiotic.
Meagher, N.; Hettiarachchi, D.; Hawkins, M. R.; Tavlian, S.; Spirkoska, V.; McVernon, J.; Carville, K. S.; Price, D. J.; Villanueva Cabezas, J. P.; Marcato, A. J.
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BackgroundThe World Health Organization has developed several global template protocols for epidemiological investigations, including for household transmission investigations (HHTIs). These investigations facilitate rapid characterisation of novel or re-emerging respiratory pathogens and support evidence-based public health actions. Beyond technical readiness, community buy-in is central to the feasibility and acceptability of HHTIs. Research is needed to determine the perceived legitimacy among the community to inform local protocol adaptation and development of implementation plans that consider community attitudes and needs. MethodsIn 2025, we conducted a convenience survey of community members living in Victoria, Australia to explore: their understanding of emerging respiratory diseases; their willingness to take part in public health surveillance activities such as HHTIs; the acceptability of clinical and epidemiological data collection and respiratory/blood sample collection as main components of HHTIs, and; participant comfort towards including their companion animals in HHTIs. ResultsWe received 282 survey responses, of which 235 were included in the analysis dataset. Compared to the general Victorian population, our participants included a higher proportion of participants who reported being female, tertiary-educated, of Aboriginal and/or Torres Strait Islander heritage, born in Australia and speaking only English at home. Participants indicated overall high levels of comfort and acceptability towards participation in HHTIs, particularly in relation to clinical and epidemiological data collection, with lesser but still high levels of comfort with providing multiple respiratory specimens in a 14-day period. Participants were least comfortable with other specimens such as urine and blood. Involving companion animals in HHTIs was similarly acceptable as human-focused components. ConclusionsDespite our survey population being non-representative of the general Victorian population, our findings provide valuable descriptive insights into the acceptability of HHTIs in Victoria, Australia from which to benchmark future local and international surveys and community engagement activities.
Ahmed, W.; Gebrewold, M.; Verhagen, R.; Koh, M.; Gazeley, J.; Levy, A.; Simpson, S.; Nolan, M.
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Wastewater surveillance (WWS) is established as a vital tool for monitoring polio and SARS-CoV-2 with potential to improve surveillance for many other infectious diseases. This study evaluated the feasibility of detecting measles virus (MeV) RNA in wastewater as part of a national WS preparedness trial in Brisbane, Australia, from March to June 2025. Composite and passive sampling methods were employed in parallel at three wastewater treatment plants serving populations between 230,000 and 584,000. Nucleic acids were extracted and analyzed using RT-qPCR targeting MeV N and M genes to distinguish wild-type and vaccine strains. MeV RNA were detected in both 24-hour composite and passive samples on May 26 to 27, 2025 from the largest catchment of 584,000 which also included an international airport. No measles cases were reported in this city or region within 4 weeks of the WS detections. These were confirmed as vaccine-derived measles virus (MeVV) strain via specific RT-qPCR assay. Extraction recoveries varied (11.5% to 70.5%), with passive sampling showing higher efficiency. This is the first report of use of passive samples for detection of MeV. These findings are consistent with other studies reporting WWS results of both MeVV genotype A and wild type genotype B and/or D. It demonstrates the potential for sensitive MeV WWS with rapid differentiation of MeVV from wild type MeV shedding, including in airport transport hubs and with different sample types. Use of WWS could strengthen measles surveillance by enabling rapid detection of MeV RNA and supporting outbreak preparedness and response. This requires optimised methods which are specific to or differentiate wild-type MeV from MeVV. Furthermore, the successful detection of MeV using passive sampling in this study highlights its potential for deployment in diverse global contexts which may include non-sewered settings.
Smith, S. A. M.; Rockett, R. J.; Oftadeh, S.; Tam, K. K.-G.; Payne, M.; Golubchik, T.; Sintchenko, V.
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Streptococcus pneumoniae is the leading cause of empyema and pneumonia in children, and monitoring of effectiveness of polyvalent pneumococcal vaccines has been essential for controlling invasive pneumococcal disease (IPD) in children and elderly adults. Conventional serotyping of pneumococci has relied on Quellung reaction following laboratory culture, however more recently whole genome sequencing (WGS) has been implemented in many reference laboratories to enhance traditional typing. Pleural fluid samples from cases with empyema are often culture negative, limiting the utility of WGS and requiring polymerase chain reaction (PCR) or 16S rRNA sequencing to detect S. pneumoniae. These molecular methods have limited sensitivity and capacity to characterise pneumococcus in clinical samples, especially in specimens with a low pathogen abundance. This study applied capture-based enrichment (tNGS) to identify and characterise S. pneumoniae directly from pleural fluid samples. A total of 51 pleural fluid samples were subjected to tNGS with a custom probe panel, for 39 known positive fluids collected from IPD cases between 2018-2025 in New South Wales, Australia. tNGS results were benchmarked against molecular-based serotyping. Our tNGS achieved 100% sensitivity and specificity in detecting S. pneumoniae. Serotyping results were concordant with PCR and 95% (37/39) of S. pneumoniae PCR positive pleural fluid cases could be serotyped using tNGS. Standard molecular methods however could only determine serotype in 56% (22/39) of samples. This tNGS enabled 39% improvement in ability to directly identify and serotype IPD-associated serotypes of S. pneumoniae in difficult-to-culture pleural fluids can significantly enhance laboratory surveillance of IPD as well as our understanding of vaccine effectiveness.
Bagi, A.; Tiwari, A.; Mbachu, C. C.; Shea, D.; Tran, T. T.; Tahita, C.; Lompo, P.; Mkama, P.; Lyimo, E.; Baraka, V.; Le Tressoler, A.; Krolicka, A.
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Mobile laboratories (MLs), whether vehicle mounted or portable, provide a versatile platform for on-site wastewater and environmental surveillance (WES) of pathogens, particularly in remote locations with limited laboratory infrastructure. However, molecular workflows intended for ML deployment require careful optimization to account for locally available equipment, consumables, infrastructure, workforce capacity, and operational constraints. In this study, we optimized an integrated ML workflow combining Oxford Nanopore Technologies (ONT) for shotgun metagenomics, multiplex metabarcoding for community level microbial analysis, and Biomeme based qPCR for targeted pathogen analysis. To further explore the potential of metagenomics for resistome assessment, we evaluated two whole metagenome enrichment approaches for their ability to improve detection of antimicrobial resistance genes. We introduce and validate a novel ONT based strategy for multiplexed sequencing small subunit (SSU) rRNA amplicon sequencing, enabling simultaneous profiling of bacteria, archaea, and microeukaryotes in complex microbial communities with multiplex metabarcoding. Sample pretreatment and nucleic acid (NA) extraction in this ML workflow were optimized using a combination chemical mechanical lysis approach followed by magnetic bead based NA purification. Workflow performance was verified using a mock community (ZymoBIOMICS Microbial Community Standard, Zymo Research, USA) and wastewater samples spiked with inactivated Mpox virus (MPXV), demonstrating accurate taxonomic representation and sensitive MPXV detection. Comparison with a commercial ZymoBIO bead beating kit for sediment sample showed ML NA extraction performed comparably. The time efficient multiplex metabarcoding workflow enabled simultaneous profiling of bacterial, archaeal, and eukaryotic diversity and produced results more concordant with qPCR based pathogen detection than the REPLIg Cell Whole Genome Amplification (WGA) & Whole Transcriptome amplification (WTA). The protocol for Mpox virus genome characterization was successfully validated for whole genome sequencing (WES) based detection and incorporated into the standard ML workflow. Across both high and low biomass environmental matrices, the Multiple Displacement Amplification (MDA) based metagenomic workflow, combined with the ML NA extraction procedure, reliably reproduced the expected composition of the Microbial Community Standard. Collectively, the integration of ONT technology with MDA metagenomics and mobile qPCR workflows provides an effective One Health approach for pathogen surveillance and outbreak response across heterogeneous environmental settings, which was later further enhanced by an offline bioinformatic and visualization pipeline enabling near real time detection of pathogens and AMR thus early risk assessment.
Itani, D.; Smaoui, H.; Thabet, L.; Zribi, M.; Dhraief, S.; Kanzari, L.; Meftah, K.; Achour, W.; Baker, D. J.; Moss, C.-J.; Philips, L. T.; Foster-Nyarko, E.; Boutiba-Ben Boubaker, I.; Holt, K. E.
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Third-generation cephalosporin (3GC)-resistant Klebsiella pneumoniae are an increasing public health threat in Tunisia, yet there is limited data on the circulating lineages and antimicrobial resistance (AMR) determinants underlying this threat. Here, we employed whole-genome sequencing (WGS) in the Tunisian AMR surveillance system (TARSS) to characterize the 3GC resistance mechanisms, population structure, virulence, and transmission across three participating sentinel hospitals in Tunis and Ben Arous. We sequenced a balanced sample of stored 3GC-resistant (3GCR) isolates from blood and urine collected between 2018 and 2022. Of 322 sequenced isolates, 286 (89%) were confirmed as K. pneumoniae, representing 28.5% of all stored 3GC-resistant isolates. The population structure was diverse (68 sublineages) and distinct between hospitals, although several globally distributed sublineages were detected across sites (SL383, SL101, SL307, SL15). Extended-spectrum {beta}-lactamases (ESBL) genes were detected in 77% of genomes, with blaCTX-M-15 (65.4%) and blaCTX-M-14 (8%) dominant at all sites and across diverse sublineages. AmpC genes occurred in 9%, and carbapenemase in 19.6% (blaOXA-48, 14.7%; blaNDM-5, 4.5%; blaNDM-1, 3.8%), with carbapenemases mainly observed amongst SL147 and SL383 at Hospital B (41.7%). Despite sequencing less than a third of the unique 3GCR infections in each hospital, we identified 24 probable nosocomial transmission clusters involving 64 isolates. Each cluster was restricted to a single hospital, although many were detected across multiple wards in the same hospital. The acquired virulence-associated locus (ICEKp) encoding yersiniabactin was common (48.6%). Hypervirulence-associated markers (encoding aerobactin, salmochelin, and/or hypermucoidy) were rare (8.7%) but increasing over time. These were mostly found in sublineages in which convergence of ESBL and hypervirulence has been reported in other settings (including SL147, SL101 and SL383), suggesting international dissemination of convergent strains. These findings show sustained ward-level nosocomial transmission of 3GCR K. pneumoniae lineages and site-specific differences in ESBL and carbapenemase burdens, which call for targeted infection prevention and control and for future routine integration of WGS into TARSS.
Tiseo, K.; Dräger, S.; Santhosh Kumar, H.; Alkhazashvili, M.; Hammann, A.; Risch, P.; Willi, R.; Mkhatvari, T.; Fialova, C.; Adlhart, C.; Szabo, D.; Suknidze, M.; Patchkoria, I.; Broger, T.; Ivanova Reipold, E.; Varshanidze, K.; Osthoff, M.
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1.Etiological diagnosis of lower respiratory tract infections (LRTIs) relies on sputum or bronchoalveolar lavage (BAL), which may be difficult to obtain or invasive. Exhaled breath aerosol (XBA) sampling offers a non-invasive alternative for pathogen detection. We evaluated the performance of the AveloMask, a face mask-based device designed to capture XBAs for molecular testing. In this prospective paired-sample study, hospitalized adults with pneumonia at three hospitals in Switzerland and Georgia provided an XBA sample using the AveloMask and a lower respiratory tract (LRT) specimen (sputum or BAL). XBA samples were analyzed by multiplex PCR using the Roche LightMix(R) panel and LRT samples were tested using the BioFire(R) FilmArray(R) Pneumonia Panel. Concordance between XBA and LRT samples was assessed using positive percent agreement (PPA), negative percent agreement (NPA), and overall percent agreement (OPA). Ninety-three participants were enrolled and 63 participants provided paired samples. AveloMask sampling identified the dominant pathogen (lowest Ct value in the LRT sample) in 40/47 LRT-positive cases (85.1%). Across all targets, PPA was 61% (95%CI, 50-72%), NPA was 100% (95%CI, 99-100%), and OPA was 95% (95% CI, 92-96%). PPA was higher for bacteria than for viruses and lower PPA was largely driven by reduced detection of low-abundance or co-infecting pathogens. In a subset analysis, AveloMask results showed substantial overlap with standard-of-care testing and could have supported antimicrobial de-escalation. Breath aerosol sampling using the AveloMask enabled non-invasive molecular detection of LRT pathogens in pneumonia cases and may complement conventional standard-of-care testing, particularly when sputum is unavailable.
Young, I.; Jardine, R.; Desta, B. D.; Edge, T. A.; Saleem, F.; Pearl, D. L.; Majowicz, S. E.; Brooks, T.; Nesbitt, A.; Sanchez, J. J.; Schellhorn, H. E.; Elton, S.; Schwandt, M.; Lyng, D.; Krupa, B.; Montgomery, E.; Patel, M.; Tustin, J.
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Background: Beaches are popular summertime destinations in Canada. However, they can be affected by specific fecal pollution sources, increasing the risk of recreational water illness. Objectives: This study was conducted to determine the risks of acute gastrointestinal illness (AGI) among Canadian beachgoers and to evaluate the influence of different fecal indicator bacteria (FIB) and other water quality measures on assessing these risks. Methods: In a prospective cohort design, beachgoers were recruited at sites across Canada from 2023 to 2025. Sociodemographic characteristics and exposures were determined through an on-site survey, with a 7-day follow-up survey to determine risks of AGI. Bayesian mixed-effects logistic regression models were fitted to evaluate the effects of an ordinal water contact variable (no contact, minimal contact, body immersion, and swallowed water) on the incident risk of AGI, with an interaction included for water quality indicators. The levels of six FIB and water quality measures were assessed: Escherichia coli, enterococci DNA, three microbial source tracking DNA markers (human HF183/BacR287, human mitochondria, seagull Gull4), and turbidity. Results: A total of 4085 participants were recruited, with 67.6% completing the follow-up survey. The overall incident risk of AGI was 2.6%. Both swallowing water and body immersion increased AGI risks compared to no water contact: median of 20 excess cases (95% Credible Interval [CrI]: 4, 64) and 5 excess cases (95% CrI: 1, 19) of AGI predicted per 1000 beachgoers, respectively. Escherichia coli and seagull DNA marker levels were associated with AGI among those who had water contact, particularly among those who reported swallowing water. Discussion: While the overall burden of AGI due to beach water contact in Canada was low, increased risks are associated with E. coli levels particularly among those who swallow water. This could be related to fecal contamination from seagulls. However, there is substantial uncertainty in the predicted effect sizes.
Jafarifiroozabadi, R.
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Background: Safety is a critical concern in behavioral health crisis units (BHCUs), where environmental risks (e.g., ligature points) can lead to injury to self or others. However, limited research has examined how perceived safety influences facility selection among patients and care partners, or how these perceptions align with AI-driven safety risk assessments in such environments. Method: To address these gaps, a nationwide discrete choice online survey was conducted using image-based scenarios of BHCU environments, where participants selected preferred facilities based on a range of attributes, including environmental safety risks (e.g., ligature points). Additionally, participants identified safety risks in survey images, which were compared with outputs from an AI-driven tool developed and trained to detect environmental risks by experts. Quantitative analysis using conditional logit models examined the influence of attributes on facility choice, while spatial comparisons of annotated images and heatmaps assessed participant and AI-identified risk alignments. Results: Findings revealed that the higher frequency of safety risks in images significantly reduced the likelihood of facility selection (p < .001, OR {approx} 1.28), highlighting the importance of perceived safety in user decision-making. While there was notable alignment between heatmaps generated by participants and AI, key differences emerged, suggesting that participant safety perception was influenced by features not fully captured by AI, such as the type of materials or unknown, out-of-label safety risks in facility images. Conclusions: Despite these limitations, results highlighted the value of integrating AI-driven assistive tools for non-expert user safety risk assessment to support decision-making for safer BHCU environments.
TANKPINOU ZOUMENOU, H.; Faucher, J.-F.
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Background: Metronidazole (MTZ) is a first-line antibiotic for several enteric infections. Its use is common in low-income countries, where most primary-care consultations are conducted by nurses. However, increasing resistance among some enteric pathogens is a growing concern. Using WHO guidelines, we conducted a register-based cross-sectional study to assess MTZ prescribing practices and their determinants in public and private primary healthcare facilities in South Benin. Methods: We performed a register-based cross-sectional study covering the year 2020 in 11 primary healthcare facilities (5 public and 6 private) in Abomey-Calavi, South Benin, following WHO recommendations. In total, 200 visits per facility were selected using systematic random sampling. The primary outcome was the prevalence of MTZ prescription. Determinants of MTZ prescription were identified using multivariable logistic regression analysis. Results: In total, 2,200 medical visits were analyzed. The median age of patients was 19 years, and 57% were female. Antimalarials were prescribed in 52% of visits. Antibacterial agents were prescribed in the majority of visits, with MTZ being the second most frequently prescribed antibiotic (18%), after aminopenicillins (27%). In multivariable analysis, digestive symptoms (adjusted odds ratio [aOR], 8.65; 95% confidence interval [CI], 6.49-11.6), genitourinary symptoms (aOR, 6.84; 95% CI, 3.18-15.0), and skin lesions (aOR, 2.39; 95% CI, 1.58-3.60) were independently associated with increased odds of MTZ prescription. In contrast, fever (aOR, 0.66; 95% CI, 0.49-0.87), respiratory symptoms (aOR, 0.44; 95% CI, 0.26-0.71), and malaria (aOR, 0.21; 95% CI, 0.15-0.28) were associated with decreased odds. Visits in the private sector were also associated with higher odds of MTZ prescription compared with the public sector (aOR, 2.31; 95% CI, 1.78-3.02). Conclusion: MTZ is the second most commonly prescribed antibiotic in primary care in the study area, with its use largely driven by digestive symptoms. Further studies are needed to assess the appropriateness of this prescription. Additionally, research is warranted to understand better the determinants of higher antimicrobial prescribing in the private healthcare sector.