Journal of Global Antimicrobial Resistance

Rationale: Infections due to Extended-spectrum {beta}-lactamases-producing Enterobacterales (ESBL-PE) require empirical treatment with carbapenems. ESBL-PE carriage is considered as a risk factor for ESBL-PE involvement during ICU infection. Our aim was to determine factors that may predict the actual involvement of ESBL-PE. Methods: A two-periods bicentric ambispective study including ICU ESBL-PE carriers patients from April 2011 to January 2019. All ESBL-PE carriers who developed an infection were analyzed. Results: 6112 patients and 4902 patients were screened during the two periods. 384 and 232 ESBL-PE carriers were identified. Total number of infectious episodes were 146 and 114, respectively. A total of 144 pneumonias, 42 urinary tract infection and 45 digestive infections were studied. An ESBL-PE was involved in 35 (24.3%) episodes of pneumonia, and 44 (37.9%) of extra-pulmonary infections. The most frequent ESBL-PE involved were K. pneumoniae, E. cloacae and E. coli. Similar species and phenotypes were present in colonisation and infection in 29 (82.8%) of pneumonia and in 40 (90.9%) of extra-respiratory infection. Multivariate analysis identified Klebsiella pneumonia or Enterobacter cloacae carriage as risk factor for ESBL-PE involvement in pneumonia and E. coli carriage and detection of ESBL-PE carriage before ICU admission as protective factors. Conclusion: In our study an ESBL-PE involvement is infrequent in pneumonia. A known carriage before ICU admission and E. coli carriage are factors associated with the absence of ESBL-PE un the episode of respiratory infection. A confirmation of our findings could lead to a reduction in the empirical use of carbapenems in this population.

Background: Antimicrobial resistance (AMR) disproportionately affects low- and middle-income countries (LMICs) such as Pakistan, where obstetric and gynaecological (OBGYN) patients carry high antibiotic exposure. Specialty-specific drug utilization data with concurrent stewardship audit remain scarce. This study evaluated antibiotic prescribing patterns, consumption metrics, and antimicrobial stewardship program (AMS) compliance in OBGYN inpatients at a public sector tertiary care hospital. Methods: A prospective cross-sectional study was conducted in OBGYN wards of Dow University Hospital, Karachi, from 1 September to 31 October 2025. Women receiving [≥]1 systemic antibiotic were included. Daily AMS rounds were conducted by an Infectious Diseases physician and pharmacist. Antibiotic consumption was measured as Defined Daily Doses (DDD) and Days of Therapy (DOT) per 1,000 patient-days (total = 821). Antibiotics were classified by WHO AWaRe (2023) framework. Results: Of 812 total admissions, 278 patients (34.2%) received [≥]1 antibiotic and were enrolled (205 obstetric, 73 gynaecological), generating 636 prescriptions (mean 2.29/patient). Surgical prophylaxis was the predominant documented indication (213, 33.5%); 65.1% carried no documented indication. By AWaRe classification, 53.6% were Access-group and 46.1% Watch-group. Ceftriaxone (38.4%) and metronidazole (36.8%) together represented 75.2% of prescriptions. Combined DDD/1,000 patient-days was 1,758.6 and DOT/1,000 patient-days was 1,852.7. AMS compliance was 0%. Conclusions: This study documents high antibiotic prescribing burden, near-universal documentation failure, and zero AMS compliance in OBGYN inpatients at a Pakistani public sector hospital. The predominance of Watch-group antibiotics and undocumented surgical prophylaxis highlights structural stewardship gaps. Findings support urgent need for institutional OBGYN antibiotic guidelines and structured pharmacist-led AMS programs.

Background: Multidrug-resistant tuberculosis (MDR-TB) remains a significant public health threat in low- and middle-income countries, including Cameroon. This systematic review and meta-analysis aimed to determine the pooled prevalence of MDR-TB and other specific anti-tuberculosis drug resistance patterns, as well as to identify factors associated with drug-resistant tuberculosis in Cameroon. Methods: A comprehensive literature search was conducted in PubMed, Scopus, Web of Science, Embase, Cochrane Library, and African Journals Online. Additional studies were identified through Google Scholar and reference list screening. Observational studies (cross-sectional, cohort, and case-control) reporting drug resistance among bacteriologically confirmed tuberculosis patients in Cameroon were eligible. Joanna Briggs Institute critical appraisal tools were used to critically assessed the study quality. Pooled prevalence estimates were calculated using random-effects meta-analysis. Subgroup analyses and meta-regression explored sources of heterogeneity. A p-value 0.05 was considered statistically significant. Results: Twenty-eight studies conducted between 1995 and 2022 were included. The pooled prevalence of MDR-TB was 5.2% (95% CI: 2.7-9.6; 21 studies; n = 7,515), with significantly higher acquired resistance (11.6%; 95% CI: 6.3-20.3) than initial resistance (2.0%; 95% CI: 1.1-3.5). The highest pooled MDR-TB prevalence was observed in the most recent studies (38.8%; 95% CI: 33.7-44.2), and the lowest in 2015-2019 (2.7%; 95% CI: 0.4-15.2). Any resistance to anti-tuberculosis drugs was 16.0% (95% CI: 10.3-23.9; 28 studies; n = 9,931), and rifampicin resistance was 4.6% (95% CI: 2.4-8.6; 25 studies; n = 8,728). Monoresistance was highest for streptomycin (6.4%; 95% CI: 3.7-10.8) and isoniazid (4.7%; 95% CI: 3.0-7.4). Previous tuberculosis infection was the strongest predictor of drug resistance (OR = 3.9; 95% CI: 1.8-8.4), followed by alcohol consumption (OR = 1.8; 95% CI: 1.2-2.7) and history of incarceration (OR = 1.7; 95% CI: 1.1-2.6). High heterogeneity was observed across most of the pooled estimates. Conclusions: Drug-resistant tuberculosis, particularly MDR-TB, poses a substantial burden in Cameroon, with acquired resistance significantly exceeding initial resistance. Previous tuberculosis infection, alcohol use, and incarceration are key modifiable risk factors. These findings underscore the urgent need to strengthen routine drug susceptibility testing, scale up rapid molecular diagnostics, enhance treatment adherence strategies, and implement targeted interventions for high-risk populations.

Oligella otitidis MSHR-50489EDL strain (ATCC: TSD462; DSMZ: DSM118617) is a new species of the genus Oligella that was isolated from a middle ear discharge swab from a child with chronic suppurative otitis media (CSOM). This Gram-negative coccobacillus produces small, circular, smooth, whitish-opaque and occasionally mucoid colonies. It grows in aerobic conditions at a temperature range from 25-42oC. Phylogenetic analysis demonstrates a relationship to other species of the genera Oligella and average nucleotide identity and digital DNA/DNA hybridization values indicate a distinct species in comparison to other Oligella species. Thus far, the majority of isolates exhibit resistance to ciprofloxacin, the first line treatment for CSOM.

IntroductionEnteric fever, being endemic with seasonal peaks in low- and middle-income countries, is a major health concern. Moreover, the rise in antibiotic resistance has exacerbated the situation. This study was undertaken to investigate the lytic bacteriophages against Salmonella Typhi with a potential for phage therapy. Materials and MethodsA hospital-based cross-sectional study was conducted from October 2023 to March 2024. Blood cultures were processed by the BACTEC automated culture system following standard microbiological techniques to isolate typhoidal Salmonella. Antibiotic susceptibility was tested by the modified Kirby-Bauer disc diffusion method. Lytic bacteriophages isolated by the double-layer agar method were assessed for their host range and lytic ability with spot and turbidimetric assays. ResultsOf the total 1054 blood specimens, 35 (3.2%) were positive for S. Typhi. All the isolates were susceptible to first-line antibiotics--ampicillin, chloramphenicol, and cotrimoxazole. The isolates were also sensitive to nalidixic acid (80%) as well as fluoroquinolones; ciprofloxacin (62.86%), levofloxacin (77.14%), and ofloxacin (80%). Fifteen lytic phages were isolated against S. Typhi Ty2 and CT18 strains. Four phages--vB_SaTy_ST2, vB_SaTy_ST7, vB_SaTy_ST17, and vB_SaTy_ST18--lysed all 35 clinical S. Typhi isolates. While vB_SaTy_ST17 and vB_SaTy_ST18 also lysed 7 out of 20 S. Paratyphi A isolates. Three phages (vB_SaTy_ST2, vB_SaTy_ST7, vB_SaTy_ST17) were tested against S. Typhi isolate S30. Individually, vB_SaTy_ST17 suppressed the growth for 13 hours, vB_SaTy_ST2 and vB_SaTy_ST7 for 10 hours. The phage cocktail vB_SaTy_ST2 + vB_SaTy_ST17 was the most effective, which extended the inhibition time to 15 hours. ConclusionThis study highlights the ongoing burden of enteric fever in Nepal and the increase in susceptibility of S. Typhi to nalidixic acid and fluoroquinolones. It also demonstrates the promising lytic potential of bacteriophages, particularly vB_SaTy_ST17 and the phage cocktail vB_SaTy_ST2 + vB_SaTy_ST17, against clinical S. Typhi, highlighting their potential as alternatives to antibiotics.

Background Klebsiella pneumoniae (Kpn) is a major cause of neonatal sepsis in Africa. 3rd generation cephalosporin and gentamicin resistant Kpn is the norm in many sites, rendering WHO recommended first- and second-line antimicrobials ineffective. An understanding of which neonates and infants are most at risk of sepsis caused by Kpn would support the case for improved access to WHO watch and reserve antimicrobials (i.e. carbapenems) for patients most likely to benefit from them. Methods A prospective case-control study was conducted at Queen Elizabeth Central Hospital, Malawi. Cases were infants <3 months of age with blood or CSF culture confirmed Kpn infection. Controls were healthy infants from the same wards and were matched 2:1. Univariate and multivariate logistic regression were performed on mean-centred data to determine risk factors for infection with Kpn. Results We analysed data from 38 cases and 76 controls between August 2021 and April 2023. Mortality at 3 months of age was 21/38 (29%) for cases, with 14/38 (37%) identified postmortem and 6/76 (7.9%) for controls (OR 14.0 (95% CI 4.59, 49.2, p>0.001). Cases were more likely to be born out of QECH than controls (42% vs. 24%, p = 0.043), and cases had lower birthweights (median 2200g vs. 2850g, p = 0.005). Multivariate logistic regression analysis revealed that increasing birthweight was protective against Kpn infection (OR: 0.858 [95% CI: 0.745, 0.987] per 100g increase), while longer hospital stay was associated with increased odds of infection (OR: 1.148 [95% CI: 1.012, 1.1.303] per additional day). Most infecting isolates (34/38 [89%]) were resistant to first- and second-line antimicrobial agents, but all were sensitive to meropenem and 33/36 [92%] to amikacin. Conclusion Low birthweight infants with prolonged hospital stay were at greatest risk of Kpn infections that were typically resistant to WHO first- and second-line antimicrobial therapy. These infants should be prioritised for antibiotics that have the potential to be life-saving. The overlapping and evolving nature of these risk factors makes it difficult to design a simple tool to support empiric initiation of meropenem. Neonates critically ill with Kpn sepsis cannot, however, afford to wait for blood culture confirmation before receiving effective treatment. This highlights the need for empiric decision making frameworks that allow rapid initiation of effective therapy in high-risk neonates.

Antimicrobial resistance (AMR) exacerbates bacteraemia in cancer patients, particularly in low-resource settings. At the Uganda Cancer Institute, high rates of Enterobacterales producing extended-spectrum {beta}-lactamases (ESBLs) have been reported, with DNA-based detection of bla genes limited to PCR. This study aimed to determine whether bacterial genomic DNA shipped at ambient temperature from Uganda to the UK retained sufficient quality for whole-genome sequencing (WGS), to allow in-depth genomic analyses of isolates. Genomic DNA was extracted from Gram-negative bloodstream isolates (n=77) in Uganda and shipped to the UK at ambient temperature. rpoB gene (77/77, 100%) and WGS data (72/77, 93.5%) were generated for isolates, with 66/72 (91.7%) genomes of high-quality (Escherichia coli n=34; Klebsiella spp. n=32). Bioinformatic analyses included species identification, sequence typing, SNP analysis, AMR and virulence gene profiling, and comparison with publicly available genomes of Ugandan isolates. Phenotypic-genotypic concordance was generally high: 7/77 (9.1%) isolates were misidentified by phenotypic testing, and two showed unexplained carbapenem resistance. E. coli isolates showed diverse sequence types, with high prevalence of blaCTX-M (91.2%) and blaOXA-1 (47.1%); carbapenemase genes were rare. Klebsiella isolates lacked hypermucoidy loci and displayed diverse capsule types, with a high prevalence of ESBLs. Genomic clustering suggested limited within-hospital transmission of strains. Genomic data can provide important insights into the dissemination of bacterial subclades of global concern. The widespread AMR genotypes reported here highlight the need for improved diagnostics and updated treatment guidelines for bacteraemia in Ugandan cancer patients.

Background: Invasive pulmonary aspergillosis (IPA) is a serious infection in critically ill patients. Galactomannan detection in endotracheal aspirates (ETA) has emerged as a promising non-invasive diagnostic method. This study evaluates the supportive diagnostic value of ETA galactomannan in ICU patients suspected to have IPA. Methods: We conducted a prospective observational cohort study over two years, enrolling 120 patients in the medicine ICU at a tertiary care centre in India (January 2022 to October 2023). Patients aged over 14 years on mechanical ventilation for >48 hours meeting the entry criteria of the BM-AspICU algorithm were included. ETA galactomannan was measured and correlated with IPA classification. Results: Of 120 patients, 37% (n=44) had probable IPA and 63% (n=76) were classified as colonisers or possible IPA. The optimal ETA galactomannan cut-off was 1.097, yielding sensitivity 72.73% (95% CI 57.2 - 85.0%), specificity 84.2% (95% CI 74.4 - 90.7%), PLR 4.86, NLR 0.35, and AUC 0.844 Conclusion: ETA galactomannan supports IPA diagnosis with favourable sensitivity and specificity. However, given the limitations of clinical scoring-based reference standards and the potential plateau in colonizer reduction at higher cut-offs, it should be integrated into a comprehensive diagnostic approach incorporating clinical, radiological, and microbiological criteria.

Objectives To evaluate two-year, side-by-side outcomes of a prospective audit and feedback (PAF)-based antimicrobial stewardship program (ASP) in a quaternary ICU in Ghana, comparing diagnostic stewardship, antimicrobial prescribing patterns, and clinician adherence to stewardship recommendations between 2024 and 2025. Longitudinal PAF data from low- and middle-income countries (LMIC) quaternary ICUs are scarce; this study addresses that evidence gap. Methods A retrospective comparative analysis of routine Antimicrobial Stewardship (AMS) surveillance data was conducted at the University of Ghana Medical Centre ICU: 102 visits in 2024 and 63 in 2025. Proportions were compared by chi-square or Fishers exact test; continuous variables by Mann-Whitney U. Wilson score 95% confidence intervals (CIs) were computed for primary proportions. Results Biomarker-guided prescribing rose from 86.3% to 100% of visits (p=0.005) and culture and sensitivity testing from 74.5% to 90.5% (p=0.02). Targeted (culture-guided) therapy increased significantly from 23.5% to 41.7% of antibiotic recipients (p=0.03), while empiric prescribing declined correspondingly. Overall antibiotic utilization remained high in both years (96.1% vs 95.2%; p=1.00), and meropenem use rose from 42.9% to 56.7% (p=0.13). AMS interventions were recommended in 67.6% and 63.5% of visits, respectively. Clinician acceptance improved markedly from 40.6% (95% CI: 29.8-52.4%) to 67.5% (95% CI: 52.0-79.9%) (p=0.01). Conclusions Two years of PAF in a Ghanaian quaternary ICU demonstrated progressive program maturation: universal biomarker adoption, a significant shift toward targeted prescribing, and markedly enhanced clinician acceptance. Persistently high antibiotic utilization and rising carbapenem dependence underscore the need for sustained surveillance and carbapenem-sparing strategies in LMIC critical care.

ObjectivesFosfomycin (FOS) remains an important therapeutic option for urinary tract infections caused by uropathogenic Escherichia coli (UPEC), but specific virulence traits as biofilm formation, metabolic adaptation, and antimicrobial resistance may limit its efficacy. This study investigated whether the natural compound, trans-cinnamaldehyde (t-CA) potentiates FOS activity against UPEC and explored the underlying mechanisms of its effect MethodsThe interaction between t-CA and FOS was assessed using checkerboard assays, time-kill analysis. We evaluated biofilm viability and structure using confocal and scanning microscopy as well as catheter-associated biofilm models. Next, effects on membrane integrity, cell-surface properties, membrane potential, intracellular pyruvate levels, and resistance evolution during serial passage were evaluated. Molecular docking was used to explore potential interactions of t-CA with enzymes involved in pyruvate metabolism. Galleria mellonella infection model was employed to evaluate in vivo therapeutical efficiency. Resultst-CA potentiated FOS activity against laboratory, reference, and clinical UPEC strains, with synergistic or additive interactions observed across the tested collection. The combination enhanced bacterial killing, reduced biofilm viability and biomass, and disrupted biofilm architecture. In catheter-associated biofilms, combined treatment markedly impaired surface-associated UPEC communities. t-CA reduced extracellular matrix abundance and altered cell-surface hydrophobicity and membrane potential without inducing detectable oxidative stress. Mechanistically, t-CA affected pyruvate homeostasis, reduced intracellular pyruvate levels, and phenotypically intersected with the BtsSR pyruvate-sensing pathway. Serial exposure to FOS alone rapidly increased MIC, whereas t-CA limited this phenomenon and did not itself promote reduced susceptibility. The compounds combination also improved survival of UTI89-infected G. mellonella larvae. Conclusionst-CA enhances FOS activity against UPEC through complementing the antibiofilm and metabolic effects. By weakening biofilm matrix integrity, perturbing pyruvate homeostasis, and limiting FOS-associated MIC elevation, t-CA represents a promising adjuvant candidate for improving FOS efficacy against biofilm-associated UPEC infections.

Macrolides are key agents for treating infections caused by non-tuberculous mycobacteria (NTM). Nevertheless, chromosomal erm genes conferring inducible macrolide resistance are described in some NTM species, such as Mycobacterium abscessus and M. fortuitum, whereas M. chelonae had long been considered as lacking a functional erm. Recent descriptions from the USA and Japan of a new plasmid-borne erm(55) (erm(55)P) in M. chelonae and other rapidly growing mycobacteria (RGM) have challenged this assumption. We investigated erm(55)P occurrence in clinical RGM referred to the French National Reference Centre for Mycobacteria between 2012 and 2026 by genome screening and erm(55)P specific real-time PCR. Positive isolates underwent long-read whole genome sequencing (GridIon, Oxford Nanopore Technologies). Clarithromycin (CLR) minimum inhibitory concentration (MIC) was determined by broth microdilution (RAPMYCO and FRATMYC, Thermo Fisher) and read up to 14 days. Five clinical isolates showing inducible CLR resistance (MIC range <0.25-64 mg/L on day 3-4 and 128 - >128 mg/L on day 14) were positive for erm(55)P: one M. chelonae, three M. neoaurum, and one M. parafortuitum. erm(55)P-positive M. chelonae genomes from this and previous descriptions did not cluster together in the phylogenetic analysis of 263 genomes. The assembled plasmids showed high similarity to previously reported erm(55)-carrying plasmids, especially within the erm(55)P region. The upstream sequence of erm(55)P showed a secondary structure compatible with a possible translation attenuation mechanism. These findings document the first report of a plasmid-borne erm(55) in Europe in M. chelonae and other RGM and raise concern about the emergence of plasmid macrolide resistance in NTM.

Background : Antimicrobial resistance (AMR) is a major public health threat globally, with a disproportionate burden in sub-Saharan Africa. Faith-based health facilities provide essential healthcare services to underserved populations, yet data on antimicrobial use in these settings remain limited. Aim : To assess antimicrobial use in Christian Health Association of Malawi health facilities using the World Health Organisation core medicine use indicators. Materials and Methods : A multicentre cross-sectional study was conducted in 29 CHAM health facilities across Malawi between January 2024 and June 2025. Data were collected from facility personnel, inpatient prescriptions, and patient interviews and analysed using descriptive and inferential statistics. Results : Average availability of key antimicrobials was 33.1% (95% CI: 29.7-36.4), while customised formularies were available in 64.3% of health facilities. Among 660 prescriptions analysed, 90.3% contained an antimicrobial agent, but only 33.2% adhered to standard treatment guidelines and 43.6% were prescribed using full generic names. Facilities with pharmacy professionals were more likely to have a facility-specific formulary (84.6% vs. 46.7%, p = 0.037). Conclusion : Antimicrobial stewardship gaps remain substantial in faith-based health facilities in Malawi and across sub-Saharan Africa, highlighting the need for targeted stewardship programmes in faith-based health facilities.

The minimum inhibitory concentration (MIC) is a standard measure for describing the lowest effective dose concentration of an antimicrobial compound in clinical practice; yet, conventional assays often require a substantial amount of antimicrobial compound, limiting their use with scarce, purified agents. Here, we describe a simple and reproducible technique to evaluate the MIC for purified compounds with a limited sample size. The protocol describes the MIC steps against a bacterial strain while minimizing the use of reagents and materials. It is helpful for screening purified natural products as antimicrobial agents and in early-stage drug discovery. The protocol adapts standard microplate-based assays for two-fold dilution of the compound, ensuring their applicability in microbiological studies. The MIC value of the standard antibiotic kanamycin against Staphylococcus aureus, Vibrio fischeri, Klebsiella pneumoniae, and Escherichia coli was determined using our method, and was found to be consistent with the conventional broth microdilution method, validating its reliability. Therefore, this method offers a practical and viable solution for antimicrobial drug discovery, addressing the disparity between limited compound availability and comprehensive microbiological assessment of MIC.

Background: Biochemical, biophysical and structural characterisation of isozymes from the ubiquitous family of iron- or manganese-dependent superoxide dismutases (SodFMs) requires the purification of high-quality preparations of recombinant enzymes. Determination of their key biochemical parameter, their catalytic metal-preference, requires the comparison of the catalytic turnover of samples loaded exclusively with iron versus samples loaded exclusively with manganese. Both of these aims are inhibited by the potential contamination of recombinant preparations of SodFMs, prepared by heterologous overexpression inside Escherichia coli cells, by even low levels of endogenous SodFMs from the host, both of which show very high turnover with either manganese (E. coli MnSOD) or iron (FeSOD). To overcome this problem, we created a strain of E. coli lacking the endogenous SodFMs. Here, we characterised this E. coli BL21 (DE3) {Delta}sodA{Delta}sodB strain, determining the physiological effects of SodFM deletion and demonstrating its utility for producing recombinant SodFMs for in vitro characterisation and use. Results: Genomic analysis verified the targeted gene deletions, without off-target effects. Growth, expression, elemental analysis, and proteomic data confirmed a lack of physiological defects of the strain except for a known inability to grow on glucose, which is overcome by heterologous SodFM expression. We demonstrate the utility of the strain for the efficient production of diverse recombinant SodFMs, including highly divergent, understudied isozymes, including the ability to precisely control the metal-loading of the heterologously expressed protein. Conclusions: The E. coli strain described herein is a useful microbial cell factory for production of recombinant SodFMs, which should find widespread utility as expression host of choice, enabling more efficient production of protein for studies of the biochemical, biophysical and structural properties of this remarkable family of metalloenzymes.

Importance: Respiratory syncytial virus (RSV) hospitalization rates are highest among children <2 years of age. RSV immunization with infant monoclonal antibody or maternal vaccine is recommended to protect all U.S. infants in their first RSV season. For certain high-risk children aged 8-19 months entering their second RSV season, the monoclonal antibody nirsevimab is recommended. Little is known regarding preexisting health conditions as risk factors for RSV-associated respiratory failure in children during their second season. Objectives: To describe children admitted to the pediatric intensive care unit (PICU) for RSV during their second RSV season by preexisting health conditions, and to compare demographic and clinical characteristics across groups. Design, Setting, and Participants: Surveillance registry of children 8- <24 months old admitted to the PICU in 30 pediatric hospitals in the 2023-2024/2024-2025 RSV seasons. All children had an RSV-positive respiratory sample and received respiratory support with high flow nasal cannula, noninvasive ventilation, or invasive mechanical ventilation (IMV). Exposure: Preexisting health conditions potentially increasing risk of severe RSV disease. Main Outcomes and Measures: Patients were classified into four mutually exclusive groups by preexisting health conditions: 1) U.S. nirsevimab eligible criteria, 2) other identified RSV risk conditions (with some evidence of increased risk for severe RSV), 3) other preexisting conditions, and 4) no preexisting conditions. Patient demographic characteristics and level of respiratory support received were compared. Results: Among 574 children: 47 (8.2%) had U.S. nirsevimab eligibility criteria, 76 (13.2%) had other RSV risk conditions, 96 (16.7%) had other preexisting conditions, and 355 (61.8%) had none. A higher proportion of children with nirsevimab eligibility factors (40.4%) than those with other identified RSV risk conditions (17.1%) required IMV, which was higher than other (10.4%) or no (5.9%) preexisting health conditions (ptrend<0.001). Conclusions and Relevance: Approximately 20% of children admitted to the PICU with severe RSV were in the defined groups that met U.S. nirsevimab-eligibility criteria or that had an identified RSV risk condition associated with known risk for severe RSV. A considerable proportion of both groups of children required IMV for respiratory support. These findings may help inform future deliberations regarding U.S. second season nirsevimab-eligibility recommendations.

Background: Diabetes mellitus is an increasing public health challenge in Sierra Leone, where access to diagnosis, treatment, and long-term care remains limited. Traditional medicine continues to play a significant role in disease management; however, ethnobotanical knowledge related to diabetes remains insufficiently documented. Methods: A cross-sectional ethnobotanical survey was conducted among 40 informants, including traditional healers, herbalists, and knowledgeable community members in Waterloo, Pendembu, and Bo. Data were collected using structured questionnaires administered via Kobo Toolbox and paper-based tools. Information on medicinal plants, plant parts used, preparation methods, routes of administration, and knowledge transmission pathways was obtained. Quantitative ethnobotanical indices, including Frequency of Citation (FC), Relative Frequency of Citation (RFC), and Informant Consensus Factor (ICF), were calculated. Results: A total of 21 medicinal plant species were documented. The most frequently cited species were Moringa oleifera (FC = 9; RFC = 0.225), Vernonia amygdalina (FC = 7; RFC = 0.175), and both Cassia siberiana and Telfairia occidentalis (FC = 6; RFC = 0.150). Leaves were the most commonly utilized plant part (40.9%), and decoction was the predominant preparation method (76.2%), with oral administration accounting for 95.2% of use. The Informant Consensus Factor (ICF = 0.69) indicated a relatively high level of agreement among informants. Knowledge was primarily transmitted through apprenticeship and inherited family practices. Conclusion: Traditional medicinal plants remain an important component of diabetes management in Sierra Leone. The high level of consensus among informants and the repeated citation of specific plant species suggest structured and culturally validated therapeutic practices. The findings provide a foundation for future phytochemical and pharmacological investigations and highlight the need for documentation, preservation, and sustainable utilization of ethnobotanical knowledge.

Probiotics produce antimicrobial peptides and small molecules that are secreted into the medium. Antimicrobial activity of cell-free supernatants can be tested using various qualitative and quantitative methods. Many of these techniques employ methods which introduce uncontrolled variables, impacting reproducibility and making comparison of reported results difficult. Here, we present a simple procedure for quantitative estimation of antimicrobial activity of cell-free supernatants which overcomes drawbacks of commonly used methods.

Drug-resistant tuberculosis and the rising incidence of nontuberculous mycobacterial (NTM) infections are a growing concern that demands innovative therapeutic strategies. Despite advances in diagnostics, drug discovery, and vaccine strategies, significant gaps remain. Mycobacteriophages and their lytic enzymes offer a promising solution due to their natural abundance and diversity, host specificity and ability to disrupt complex cell envelopes and biofilms. In this study, we report the genomic and functional characterization of a V-Cluster mycobacteriophage, EniyanLRS, isolated near a hospital in Delhi and the encoded endolysins LysA and LysB. EniyanLRS features a 78.53 kbp genome with a notably low GC content (56.9%) as compared to other mycobacteriophages, and an exceptionally long Tape Measuring Protein (TMP) gene (5.97 kbp). Its genome lacks genes related to lysogeny and harbours 24 tRNAs, suggesting high translational efficiency. Phenotypically, EniyanLRS exhibits a siphovirus morphology, lytic lifecycle and infects Mycobacterium smegmatis and drug-resistant Mycobacterium fortuitum. LysA, with its lysozyme-chitinase-amidase domain architecture, did not demonstrate significant antibacterial or antibiofilm activity. Conversely, LysB, an /{beta}-hydrolase, exhibited superior in vitro esterase activity compared to previously reported LysB enzymes and showed pronounced cell wall disruption of M. smegmatis and M. fortuitum, along with considerable antibiofilm efficacy (62.77% and 41.91% inhibition, respectively). Collectively, these findings highlight the potential of EniyanLRS and its LysB enzyme as potent biocontrol agents against pathogenic mycobacteria, which can be explored to treat planktonic cells and biofilm-associated infections.

IntroductionLomentospora prolificans is a pathogenic filamentous fungus that causes disease primarily in people with severely compromised immune systems. It is pan-resistant to antifungal drugs, but the mechanism of its resistance to amphotericin B (AMB) is unknown. ObjectivesWe aimed to investigate the mechanism of resistance to AMB of L. prolificans. MethodsThe AMB susceptibility of L. prolificans protoplasts was measured using broth microdilution. L. prolificans, either intact, homogenized or fractionated was incubated with AMB in broth. The same activity was carried out with Aspergillus fumigatus as a control. This broth was then used to prepare microdilution plates with Saccharomyces cerevisiae to determine the activity of the conditioned AMB. ResultsAMB was 16-fold more effective in inhibiting the growth of L. prolificans protoplasts than conidia, but only two-fold more effective against A. fumigatus protoplasts than conidia. Incubation of L. prolificans hyphae with AMB in media diminished drug activity to a much greater extent than A. fumigatus, with 8-fold greater fungal mass of the latter required to achieve the effect of the former. Homogenization and fractionization of L. prolificans revealed that the factor inhibiting AMB activity was soluble with a mass >100 kda. DNase, trypsin, proteinase K, amyloglucosidase, SDS and 0.22 m had no effect on the AMB resistance factor, while treatment with urea, acetonitrile inactivated it. ConclusionWe report a different mechanism for AMB resistance based on the existence of a substance residing in the L. prolificans cell wall that can eliminate the antifungal activity of AMB.

Background: Tuberculosis, especially drug-resistant tuberculosis (DR-TB) including multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains, remains a leading cause of infectious death worldwide. The rapid accumulation of whole-genome sequencing (WGS) data had spurred numerous computational methods for predicting antimicrobial resistance in Mycobacterium tuberculosis. However, heterogeneous datasets, preprocessing pipelines, and evaluation protocols have made fair comparisons impossible and have hindered clinical translation. A critical yet missing resource is a large-scale, unified benchmark to systematically assess and compare existing methods. Methods: We curated an integrated MTB WGS--phenotypic drug susceptibility testing (pDST) dataset from three sources: the CRyPTIC dataset (Comprehensive Resistance Prediction for Tuberculosis: an International Consortium), a published multi-study compilation, and newly curated literature-derived datasets. The final benchmark contains 54,364 paired WGS-pDST records with broad geographic, lineage, and drug coverage. After harmonizing phenotypes and generating standardized variant features, we evaluated seven models (including classical machine learning and deep learning architectures) across 18 drug-level and six clinical resistance category prediction tasks. Results: XGBoost achieved the highest mean drug-level AUPRC (0.674) and F1-score (0.620) and ranked first in AUPRC for 11 of 18 drugs, whereas WDNN achieved the highest mean AUROC. Random forest yielded the highest mean specificity (0.956) and accuracy (0.933), whereas logistic regression achieved the highest mean recall (0.774), highlighting distinct clinical trade-offs. Drug-level difficulty was highly heterogeneous: rifampicin and isoniazid were predicted robustly, whereas bedaquiline, delamanid, linezolid, and clofazimine remained persistently difficult. In clinical resistance category evaluation, RR-TB, MDR-TB, and pan-susceptibility were well predicted, but XDR-TB and other resistance categories constituted major bottlenecks. Conclusions: Under the largest unified benchmark to date, classical machine-learning methods, particularly XGBoost, provided the strongest precision--recall and F1 performance overall, while neural models remained competitive by AUROC. Emerging drugs (bedaquiline, delamanid, linezolid, clofazimine) and XDR cases remain persistently difficult to predict, identifying key bottlenecks for future method development. This benchmark can serve as a community standard for evaluating MTB resistance prediction and the provided evaluation pipeline offers an actionable baseline for regulatory qualification and clinical decision support system validation, accelerating the translation of WGS-based resistance prediction into practice.