The λ-Red recombination system is commonly used to investigate gene function in Klebsiella pneumoniae; however, unintended secondary genomic alterations in recombinant strains may remain undetected by locus-specific validation methods such as polymerase chain reaction and Sanger sequencing. Following rmpA mutant construction in the clinical hypervirulent K. pneumoniae (hvKp) strain K255, two recombinant derivatives showed identical results on locus-specific validation, confirming the intended rmpA replacement. However, the strains differed in uronic acid content and interaction with murine macrophages. We therefore performed whole-genome sequencing (WGS) incorporating both short- and long-read data to investigate potential secondary genomic alterations. Both recombinant derivatives harbored an additional chromosomal copy of ISKpn28 within the nlpD gene. The K255ΔrmpA-1 strain additionally exhibited a chromosomal deletion of 55,652 base pairs, whereas K255ΔrmpA-2 harbored an ISKpn74-associated plasmid inversion. These findings provide genomic evidence of ISKpn28 transposition in recombinant derivatives of the clinical hvKp strain K255, and highlight the potential value of WGS incorporating long-read sequencing for assessing genome-wide integrity in λ-Red-derived recombinant hvKp strains harboring ISKpn28.

Bloodstream infections (BSI) caused by Escherichia coli and Staphylococcus aureus are frequent and are associated with substantial mortality rate. The host immune response major determinant of the clinical outcomes during BSI. Here, we investigated the innate immune response, by characterizing peripheral blood myeloid cell subsets and caspase-1 activation as a surrogate marker of inflammasome activation in patients with E. coli or S. aureus bacteremia. We prospectively enrolled patients with monomicrobial bacteremia and analyzed peripheral blood samples by flow cytometry. We observed distinct alterations in myeloid cell composition and caspase-1 activation associated with E. coli bacteremia, including an expansion in intermediate and non-classical monocytes, and increased caspase-1 activation in non-classical monocytes. In contrast, these changes were not observed in patients with S. aureus bacteremia. Additionally, we found differences in myeloid cell subset frequencies and caspase-1 activation between patients with and without sepsis, revealing distinct profiles associated with disease severity. Importantly, the differences in myeloid cell subsets and caspase-1 activation between E. coli and S. aureus bacteremia were primarily attributable to the infecting bacteria rather than disease severity. Furthermore, our findings suggest that eosinophil counts, and caspase-1 activation may serve as potential markers to distinguish between bacterial BSI and viral infections. Overall, our study highlights the importance of considering both the infecting pathogen and disease severity when studying the host immune response during bacteremia and suggests that potential innate immune biomarkers may improve the diagnosis and clinical management of BSI.

Foot-and-mouth disease (FMD) is a highly contagious virus with severe economic and animal welfare consequences if introduced into nonendemic regions. Given its broad host range, including domestic and wild animals, wildlife must be considered in FMD preparedness and response planning. We conducted a scoping review according to PRISMA-ScR guidelines and narrative synthesis of 205 publications (year 2000 onward) to examine wildlife susceptibility to FMD and associated control strategies. In the Canadian context, over a dozen native wild ungulate species, as well as feral pigs, are known or likely to be susceptible to FMD. Experimental studies confirm that species such as deer, bison, and pronghorn can develop clinical disease and transmit FMD, yet their potential role as long-term maintenance hosts remains unclear. This knowledge gap highlights the critical need for targeted research to inform surveillance and control strategies in disease-free regions. Globally, wildlife-livestock interfaces are recognized as critical points for potential FMD transmission. Surveillance typically relies on opportunistic sampling of hunted or found-dead animals. Modeling studies suggest that outbreaks in wildlife are generally self-limiting without livestock involvement, constrained by ecological factors such as seasonality, population density, and short infectious periods. However, environmental persistence and shared resource use (e.g., water and feed) may facilitate transmission. Control measures include barrier fencing and livestock ring vaccination. Wildlife culling is widely considered infeasible, environmentally damaging, and socially contentious, with use recommended only under narrow, evidence-based conditions. Enhanced biosecurity, such as reducing wildlife attractants on farms, and movement restrictions are consistently emphasized. Preparedness in nonendemic regions should prioritize risk-based wildlife surveillance, cross-sectoral coordination, response protocols, and strategies to mitigate impacts on vulnerable species. An integrated approach encompassing ecological, wildlife, and agricultural perspectives is essential for coordinated, science-based responses to future FMD incursions involving wildlife.

Refusal of blood transfusion has historically been associated with members of the Jehovah's Witness (JW) faith. As such, a range of transfusion-sparing strategies have been developed for these individuals and others who refuse standard blood transfusion. Techniques that maintain blood in continuous circuit with the patient are often acceptable and have been used in operative settings, including cell salvage and acute normovolemic hemodilution. By contrast, blood that has been collected and stored for later use has traditionally been considered unacceptable, which has limited the use of both allogeneic and autologous blood products. A March 2026 update by the governing body of Jehovah's Witnesses, The Watch Tower Bible and Tract Society of Pennsylvania, moves decisions about the storage and reinfusion of a patient's own blood into the realm of individual conscience, which may expand the use of autologous transfusion. Given this change, requests for autologous transfusion from JW patients may increase. In this letter, we offer practical guidance on balancing respect for patient autonomy and religious belief with the medical and logistical challenges of autologous blood transfusion.

Heavy metals such as copper are known water pollutants that can be extremely harmful to the environment and pose serious risks to human health and aquatic organisms. The present study aimed to determine the efficiency of Achromobacter xylosoxidans isolated from waste soil as a cost-effective biosorbent for the removal of copper (II) (Cu[2+]) ions from synthetic and real wastewater.

Gnomoniopsis comprises parasitic, endophytic, and saprophytic fungi. Since its establishment, this genus has presented considerable taxonomic challenges owing to extensive morphological overlap among species, rendering traditional classification systems that rely predominantly on phenotypic characteristics no longer applicable. To resolve the current taxonomic confusion and clarify the species diversity, phylogenetic relationships, and genomic adaptive characteristics of Gnomoniopsis, the present study was carried out. In this study, the internal transcriptional spacer (ITS), translation extension factor 1-α (tef1), and partial beta-tubulin (tub2) were analyzed and combined to observe morphological characteristics of Gnomoniopsis. The isolates were identified from chestnut (Castanea), Embelia vestita, tea (Camellia sinensis), and other hosts in Sichuan Province, as well as from Quercus species and partial leaf rot from Fujian Province. Nine new species, namely G. xiaoheijiangensis sp. nov., G. embeliae sp. nov., G. yunvfengensis sp. nov., G. quercus sp. nov., G. ledongensis sp. nov., G. yonganensis sp. nov., G. conghuaensis sp. nov., G. loropetali sp. nov., and G. qixiensis sp. nov., were obtained. Phylogenomic analysis based on 7,942 orthologous proteins' clusters revealed distinct species boundaries within Gnomoniopsis. Phylogenomics has thus effectively resolved the ambiguous phylogenetic relationships in this genus. This work identifies nine previously undescribed Gnomoniopsis species, clarifies the species diversity and conserved genomic characteristics within the genus, and lays a foundation for subsequent taxonomic analyses of Gnomoniopsis.

Synthetic biology and the emerging field of biomanufacturing provide new research concepts that should be incorporated into the training of the next generation of scientists. Key elements of synthetic biology are the standardization, modularization, and characterization of biological parts. Defined parts can be assembled into functional transcription units to pathways of multiple units, chromosomes, and/or whole genomes. Here, we highlight the importance of global exchange and state-of-the-art training in the Global South to empower research and reduce the gap between countries. Based on initial seed funding, we established a multidisciplinary bilateral exchange program between Germany and Colombia, including lecture series, experimental workshops, and bilateral exchanges in training and research. The elements are adaptable and our hands-on courses can be carried out with limited resources while covering aspects of synthetic biology. Importantly, this initiative will allow early-career researchers to build an international network, opening up opportunities in research collaboration and paving the way for greater equity.

Brucellosis, caused by Brucella species, is a major zoonosis with global health and economic burdens. Conventional diagnostics suffer from slow culture, biosafety risks, cross-reactivity, and equipment dependence. Aptamers, synthetic single-stranded oligonucleotides selected via Systematic Evolution of Ligands by Exponential Enrichment (SELEX), offer promising alternatives to antibodies. This review focuses primarily on recent advances in aptamer-based diagnostic technologies for brucellosis. The success of Brucella-specific aptamers is largely attributed to the development of whole-cell-based SELEX strategies and counter-selection against non-target bacteria, which help overcome the complexity of the Brucella outer membrane and improve target specificity. Brucella-specific aptamers such as B20 (KD = 40.179 pM), B21 (KD = 184.396 pM), and WS-6 (KD = 16.23 nM) exhibit high affinity and specificity. Integrated into biosensors, they enable detection limits as low as 1.5×10[1] CFU/mL (colorimetric) and 27 cells/mL (surface plasmon resonance). While aptamer-nanoparticle conjugates also show therapeutic potential against intracellular infections via immun4omodulation, this aspect remains secondary and requires further validation, and toxicity concerns remain. Despite progress, clinical validation, matrix interference, and regulatory hurdles persist. Future directions include multi-target aptamers, nanomaterial integration, and point-of-care translation. This review underscores the central role of aptamers in brucellosis diagnosis, with complementary therapeutic potential.

Vaccinium is one of the most economically significant genera in the Ericaceae family, with breeding efforts substantially improving yield, fruit quality, and adaptation, contributing to the widespread consumption of this berry-bearing clade. Crops within this genus are valued for their high nutritional content, including antioxidants, dietary fiber, and vitamins, as well as their culinary versatility. Health benefits, such as anti-inflammatory activity, combined with advances in breeding and cultivation that enhance flavor and yield, have further boosted consumer demand. However, the application of traditional breeding methods in Vaccinium remains challenging, mostly due to their long juvenile period, polyploidy, and the complex genomes of both cultivated and wild plants. To overcome these constraints, new breeding techniques (NBTs), such as gene editing, offer tremendous potential for developing improved varieties with traits that are difficult to achieve through traditional methods. Over the past 30 years, significant advancements in Vaccinium biotechnology have been made, particularly in the field of transgenesis. These advancements, including the ability to introduce foreign genes and edit genomes, hold great promise for overcoming the challenges posed by traditional breeding techniques. Despite these developments, the implementation of NBTs in Vaccinium remains at an earlier stage compared with other horticultural crops, primarily due to: (i) recalcitrancy to stable genetic transformation and (ii) low editing efficiencies. In this review, we provide a detailed overview of advancements in regeneration and genetic transformation in Vaccinium species, along with a perspective on the major constraints and future strategies to improve Vaccinium through biotechnological methods.

Chigger mites (larval Trombiculidae) are obligate ectoparasites, some species transmit Orientia tsutsugamushi, the causative agent of scrub typhus. Despite their public health significance in rural Southeast Asia, relationships between chiggers infesting wildlife, domestic animals and those present in the surrounding environment remain poorly characterised. We hypothesised that agricultural-forest ecotones would support higher chigger occurrence and that murid rodents would be the principal maintenance hosts of chigger communities. We conducted ecological observations of chigger communities associated with backyard chickens (n = 117), free-roaming dogs (n = 22), murid rodents (n = 96), insectivorous bats (n = 24) and environmental black-plate samples (n = 150) within a scrub typhus-endemic rural village in Nan Province, northern Thailand. Host-chigger associations were characterized using bipartite and unipartite network analyses. Twenty-one chigger species were identified, including four previously known to harbour O. tsutsugamushi: Ascoschoengastia indica, Leptotrombidium imphalum, Eutrombicula wichmanni and Walchia kritochaeta. Unfed questing chiggers occurred within the central residential area rather than being restricted to peripheral agricultural-forest interfaces as hypothesised. Murid rodents contributed the greatest chigger diversity, while backyard chickens exhibited unexpectedly high infestation prevalence (54.7%) and mean abundance. Environmental chigger detection rate was 11.3%. Eutrombicula wichmanni was recorded on free-roaming dogs, representing the first documented chigger infestation on dogs in Thailand, and a complete mitogenome was obtained from environmentally collected adults. Network analysis identified distinct rodent- and domestic animal-associated chigger communities, with the environmental chiggers linking both groups, whereas bat-associated chiggers were clearly isolated. These findings reveal complex chigger community structures with distinct ecological partitioning, highlighting the importance of domestic animals, particularly backyard chickens and free-roaming dogs (in addition to rodents) as critical components of peri-domestic transmission cycles. Future studies integrating host-chigger ecology with rickettsial pathogen screening would inform targeted disease surveillance and control programmes in rural endemic communities.

Streptococcus suis is a major swine pathogen and is considered an emerging zoonotic agent. Serotyping plays a crucial role for epidemiological investigations, pathogenicity assessment, and vaccine strategy. The present study evaluated the applicability of Fourier transform infrared (FT-IR) spectroscopy, using the IR Biotyper system, for the serotype discrimination of 94 S. suis isolates belonging to 13 distinct serotypes. Analysis of the 1300-800 cm[-1] spectral region demonstrated the highest discriminatory power. Principal component analysis (PCA)/linear discriminant analysis (LDA) showed clear separation among the tested serotypes, except for serotypes 1/2 and 2. The classifier achieved overall accuracy of 90%, which increased to 96.7% when serotypes 2 and 1/2 were considered as a single category. These findings demonstrate that FT-IR spectroscopy represents a promising, rapid, and cost-effective tool for S. suis serotype discrimination. Its performance, ease of use, and compatibility with machine learning approaches support its potential implementation in routine diagnostic and epidemiological workflows. As previously reported for other clinically relevant bacteria, FT-IR spectroscopy may serve as a valuable complementary method to molecular typing, particularly in settings requiring rapid decision-making and high-throughput analysis.

Persistent fever after initiating intravenous antibiotics is common in hospitalized patients with cancer and often leads to additional imaging and antimicrobial escalation. Early identification of patients unlikely to remain febrile could provide additional information during clinical reassessment. We developed and externally validated machine learning models to predict persistent fever 48-72 h after antibiotic initiation using electronic health records from two academic hospitals. Adult cancer patients with fever within 24-48 h of treatment were included. Models incorporated demographics, comorbidities, laboratory results, vital signs, and temperature patterns. The best-performing model achieved an AUROC of 0.82 (95% CI 0.79-0.84) internally and 0.78 (95% CI 0.77-0.79) externally. Temperature-related variables were the dominant predictors. Decision curve analysis demonstrated positive net benefit across much of the evaluated threshold range. Retrospective assessment suggested that approximately one quarter of antibiotic escalations and chest CT scans occurred in patients correctly predicted not to remain febrile, highlighting the potential contextual value of fever predictions. These findings indicate that short-term fever persistence can be predicted with clinically meaningful accuracy using routine data. However, clinical use should be limited to an adjunctive role rather than standalone rule-out decision-making, and prospective studies are required to evaluate safety, clinician uptake, and real-world impact.