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This paper summarizes a decade of development of a Solar-Assisted Large-Scale Cleaning System (SALSCS) aimed at mitigating urban PM2.5. This effort has led to the construction and operation of four SALSCS units located in Xi'an (China), Yancheng (China), and New Delhi (India). Six papers have been published to document the modeling, design, construction, operation, and measurement of three generations of SALSCSs. The Weather Research and Forecasting (WRF) model was utilized to obtain local meteorological information and solar intensity conditions around the SALSCS. Reynolds-Averaged Navier-Stokes (RANS) simulations and Large Eddy Simulation (LES) have been employed to study the flow patterns and clean air concentration profiles near the units. The first generation SALSCS (Xi'an) takes the form of an updraft solar tower that utilizes solar heating to drive a large volume of air flow through the SALSCS. Filters are positioned along the flow path to remove PM2.5, resulting in cleaner air exiting the top of the tower. In the second generation SALSCS (Yancheng), the media filters are replaced with water spray to scrub out PM2.5. The third generation SALSCS (New Delhi) employs a set of fans to draw PM2.5 from the tower inlet through prefilters and final filters, blowing the cleaned air out near ground level to pedestrians surrounding the SALSCS. A proposal is presented that combines the 1st and 2nd Generation SALSCSs equipped with solar panels and direct air capture (DAC) of CO2 to achieve energy self-sufficiency and large-scale capture of 100 million tons of CO2 annually (100 Mt CO2/yr).

SUMMARYRickettsial diseases, encompassing scrub typhus, spotted fever group rickettsioses, and typhus group rickettsioses, represent a significant and escalating public health threat worldwide, particularly in the Asia-Pacific and sub-Saharan African regions. Despite their high morbidity and potential for fatal outcomes if left untreated, these infections remain notoriously underdiagnosed due to their nonspecific clinical presentation, which frequently overlaps with other acute undifferentiated febrile illnesses (AUFIs) such as dengue, malaria, and leptospirosis. This review evaluates the evolving diagnostic landscape, highlighting the severe limitations of conventional methods: the Weil-Felix test lacks necessary specificity, while the gold standard indirect immunofluorescence assay (IFA) is primarily retrospective due to delayed seroconversion. Molecular diagnostics, particularly multiplex polymerase chain reaction (mPCR), have emerged as a critical advancement, enabling early, species-specific identification during the acute phase of infection when doxycycline therapy is most effective. We further explore the paradigm shift toward syndromic molecular panels, such as the TaqMan Array Card (TAC), which facilitate simultaneous screening for multiple AUFI pathogens. Emerging platforms, including digital PCR (dPCR) for absolute quantification and CRISPR-Cas-based point-of-care (POC) systems (SHERLOCK and DETECTR), offer promising solutions for low-resource settings. Finally, this review underscores the necessity of integrating molecular surveillance within a One Health framework and utilizing artificial intelligence (AI) to address technical and implementation barriers. Overcoming these challenges is essential for transforming rickettsial diagnosis from a reactive to a proactive strategy, ultimately reducing the global burden of these neglected zoonoses.

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Scrub typhus is a notable zoonosis, yet natural host-associated pathogen prevalence and genotypic diversity remain underexplored. A total of 261 small mammals comprising rodents and shrews captured in Uttar Pradesh, India, were screened for Orientia tsutsugamushi (OT) using nested PCR. The sample included Rattus rattus (n = 28), Rattus norvegicus (n = 42), Rattus tanezumi (n = 42), Bandicota bengalensis (n = 10), Mus musculus (n = 15), and Suncus murinus (n = 124). The overall molecular prevalence was 11.9% (95% CI: 8.2-16.4) with detection rates of 13.1% in rodents and 12.1% in shrews. Prevalence was significantly higher in rural (15.2%) than urban (1.6%) settings, and during monsoon (29.3%) and post-monsoon (11.9%) periods. Phylogenetic analysis of the partial tsa56 gene identified 4 OT strains circulating in rodent and shrew hosts, including Gilliam and TA678, along with novel JJOtsu5 and JJOtsu7. Population genetic analysis revealed substantial tsa56 nucleotide and haplotype diversity under purifying selection, while recombination analysis detected recombination in a subset of sequences, also contributing to genetic diversity. Deduced amino acid sequence analysis of the contiguous partial tsa56 gene revealed distinct mutations and structural variation in Gilliam-like and TA678-like strains, while JJOtsu5 and JJOtsu7 showed no variation relative to reference strains. In silico immunoinformatic analysis predicted 8 CD8 and 55 CD4 T-cell epitopes (TCEs) eliciting a human immune response, predominantly located within the spacer region S-VDIII/IV. The CD8 epitope AQLYKDLVKL was conserved across Gilliam, TA678-like (variant-27S), and JJOtsu7 strains; while the CD4 epitope PVKVLSDKITQIYSD was shared among Gilliam, JJOtsu5 and JJOtsu7 strains with a single S290R substitution. These strains exhibited distinct histopathological responses in OT PCR-positive animal tissues. Gilliam infections caused moderate lesions in liver, spleen, and lungs; TA678 infections exhibited mild pulmonary and renal changes; JJOtsu7 induced severe pulmonary and myocardial changes; and JJOtsu5 caused mild-to-moderate inflammation in liver, spleen, and kidneys. These findings highlight substantial genetic diversity, differential immunogenic potential and variable pathogenicity among rodent and shrew associated OT strains, underscoring the need for extended surveillance and strain-specific virulence characterization for improved control strategies.

Occupational radiation exposure remains a critical concern in modern health care due to cumulative health risks for procedural staff, stringent regulatory requirements, and the proliferation of advanced imaging technologies. This systematic review aimed to evaluate occupational radiation exposure among medical personnel during standard and complex endovascular aortic repair (EVAR) procedures, identify key determinants of radiation dose, assess the efficacy of protection strategies, and propose evidence-based recommendations to optimize staff safety in endovascular practice. A systematic review was conducted following PRISMA guidelines, searching MEDLINE, Cochrane Library, and Scopus databases from January 2019 to May 2025. Eligible studies reported quantitative dosimetric data for operating room staff during elective EVAR procedures, excluding ruptured aneurysm cases and non-English publications. Study selection and quality assessment was performed by 2 independent reviewers using ROBINS-I tool. From 942 screened studies, 18 met inclusion criteria, revealing significant occupational intraoperative radiation exposure variability. Eligible studies reported dosimetric data in terms of absorbed dose (D), effective dose (ED), personal dose equivalent Hp(10) for whole-body exposure, Hp(3) for eye lens dose, Hp(0.07) for skin and extremities dose, as well as peak skin dose (PSD). The ranges of D, ED, Hp(10), Hp(3), Hp(0.07), and PSD were 0.02 to 0.12 μGy, 0.40 to 4350.00 μSv, 0.00 to 6112.70 μSv, 0.25 to 2500.00 μSv, 0.20 to 971.00 μSv, and 190.00 to 8430.00 μGy, respectively. Operator exposure patterns showed consistent left-sided predominance (left eye 200.00 vs right eye 30.00 μSv; left hand 76.90 vs right hand 46.70 μSv). Advanced imaging technologies demonstrated substantial protective benefits: 3D image fusion reduced operator ED by approximately 55% to 66%, while dual fluoroscopy with digital zooming decreased median doses by 62%. Suspended lead shielding systems eliminated axillary exposure in some cases. Supporting staff received lower but non-negligible exposure, with scrub nurses sustaining higher doses than circulating nurses. Complex EVAR procedures continue to pose significant occupational radiation risks despite technological advancements. The findings emphasize the need for optimized protection protocols, particularly for ocular exposure, standardized dosimetric reporting practices across institutions, and continued development of low-radiation or radiation-free navigation technologies. Future research should prioritize multicenter prospective studies with standardized metrics to evaluate long-term health outcomes among surgeons performing complex EVAR operations.Clinical ImpactThis systematic review offers a comprehensive synthesis of current evidence on occupational perioperative radiation exposure during minimally invasive endovascular stent grafting, providing vascular teams with insights to enhance radiation safety protocols. By analyzing dose patterns across different procedure types and technologies, the study highlights effective protective measures, incorporating newfangled imaging systems and shielding solutions. The findings support clinical decision-making by demonstrating how specific interventions can reduce staff exposure while maintaining procedural efficiency. Using an evidence-based approach, this framework guides institutions in optimizing radiation safety programs, improving staff training, and facilitating the integration of dose-reduction technologies in contemporary endovascular practice.

Scrub typhus can rapidly progress to severe disease with multi-organ dysfunction. Secondary hemophagocytic lymphohistiocytosis (HLH) is an increasingly recognized hyperinflammatory complication associated with high mortality, but adult data and simple admission-time risk tools remain limited. We conducted a single-center retrospective cohort study of hospitalized adults with scrub typhus at the First Affiliated Hospital of Guangzhou Medical University (May 2013-July 2025). Severe scrub typhus was defined by major organ involvement, shock, or in-hospital death. Variables available at admission were compared between severe and non-severe groups. Independent predictors were identified using multivariable logistic regression, and model performance was evaluated using ROC analysis. Among severe cases, HLH was identified according to standard diagnostic criteria, and clinical characteristics were compared between HLH and non-HLH patients. Among 135 patients, 44 (32.6%) developed severe scrub typhus and overall in-hospital mortality was 2.2% (3/135). Lower hemoglobin (Hb) (OR 0.965, 95% CI 0.947-0.982), higher serum creatinine (SCr) (OR 1.019, 95% CI 1.008-1.029), and lower fibrinogen (FIB) (OR 0.629, 95% CI 0.435-0.909) were independent predictors of severe disease. A parsimonious three-variable model showed good discrimination (AUC = 0.843). At the sensitivity-prioritized ROC threshold, sensitivity was 88%, specificity 46%, and the Youden index 0.34. HLH was identified in 6 patients overall, including 5 patients among the severe cases. Among severe patients, HLH cases showed higher observed in-hospital mortality than non-HLH cases (40.0% vs. 2.6%). They also had more severe thrombocytopenia and greater coagulation and organ-function abnormalities. A simple admission-time triad (Hb, SCr, and FIB) provides an interpretable tool for early identification of severe scrub typhus in hospitalized adults. HLH cases showed higher observed mortality and more severe thrombocytopenia/coagulation derangement; however, these findings should be interpreted descriptively due to the small number of cases.

Human disturbances, such as habitat destruction and overharvesting, are greatly harming ecosystems and causing significant declines in biodiversity. Although protected areas play a crucial role in conserving terrestrial mammals, nearly non-protected areas (N-PAs) have similar functions, harbor high biodiversity and ecosystem integrity, and deserve to be protected. To identify the conservation value of mammalian species in critical ecosystems within N-PAs, we conducted a camera-trap survey in Luolong County, Tibet, from November 2019 to June 2023, monitoring 159 sites and documenting 25 mammalian species across 28 similar or dissimilar habitats. We found this area was an integrity ecosystem with higher species richness and diversity in scrub and evergreen forests with notable occurrence of herbivores comprising musk deer, chinese serow, woolly hares and carnivores such as the common leopard, snow leopards, red foxes and stone marten. Mammalian species occurrence increased away from human activities. Different habitats and seasons influenced diversity and species interactions. Key findings include species preferences for specific habitats, such as blue sheep on southern slopes during snow, musk deer in mixed forests, and red foxes avoiding alpine meadows. Habitat type, elevation, and human disturbance significantly impacted species distribution and behavior. The study also found that snow leopard activity time negatively correlates with woolly hare, while common leopards are influenced positively by hares and negatively by brown bears. Red foxes are slightly more active near the chinese serow occurrence areas. Other predators and prey, such as eurasian lynx, gray wolves, musk deer and stone martens show specific seasonal and interspecific interactions, with some relationships explaining small portions of variation. Overall, species temporal detection events are interconnected through complex ecological interactions. These findings improve our understanding of habitat hosting for rare species and the balance of endangered prey and predator communities in N-PAs in Tibet, emphasizing their significance for conservation efforts.

In early 2025, OU Health identified an increase in contaminated surgical instrument sets reaching the operating room (OR). Contaminants, including bioburden, hair, holes in wrapping, debris, and wet sets, posed risks to patient safety, workflow efficiency, and OR productivity, suggesting systemic deficiencies across the sterile processing department (SPD) and perioperative workflow. We convened a multidisciplinary quality-improvement team comprising administrators, surgeons, nurses, scrub technicians, SPD technicians, and quality specialists. Using the 8-Step methodology, we conducted root cause analysis, process mapping, and longitudinal data tracking. Interventions included standardized point-of-use cleaning, enhanced SPD personal protective equipment and gowning practices, washer maintenance, adoption of heavier wrapping materials, containerization of high-risk sets, limiting tray weight, removal of static-attracting materials, and targeted staff education. From July 1, 2025, through January 31, 2026, 388 contamination events were recorded across service lines. Neurosurgery accounted for 36% (136 events) of all events. Common contamination types were holes (113 events), debris (98), bioburden (51), and filter defects (28); hair (20), wet sets (15), and improper disassembly (15) were less common but clinically significant. Temporal analysis demonstrated intermittent short-term improvements but persistent recurrence across contamination types. Contaminated sets contributed to 57% of all documented OR delays, totaling 2851 minutes. Estimated direct costs were $176 762; projected annual costs were $589 042. Contaminated surgical instrument sets represented a recurrent systems-level failure. Institution-wide efforts, including multidisciplinary collaboration, structured quality-improvement methodology, and targeted interventions, produced measurable improvements. Continued monitoring, workforce competency reinforcement, and system redesign are needed to mitigate future risk.

A laboratory-scale integrated beneficiation process was systematically developed and evaluated for the production of advanced industrial-grade, high-purity, high-whiteness silica from quartz ore sourced from the Zarrin-Cheshmeh deposit (Semnan Province, Iran). Initial characterization of the raw material (SiO₂ = 97.46 wt%) using XRF, XRD, PLM, and SEM-EDS revealed a complex mineralogical composition, comprising both free and locked impurity phases. These included lattice-bound and surface-associated contaminants such as Al₂O₃, Fe₂O₃, TiO₂, as well as clay minerals and chert fragments. A multi-stage beneficiation strategy was implemented, integrating physical pre-treatment steps (color sorting, scrubbing, shaking table separation, ultrasonic treatment, microwave treatment, and calcination) with subsequent chemical leaching stages (aqua regia at ambient temperature and 95 °C, followed by HF-HCl treatment). Color sorting effectively removed visually identifiable chert and iron-stained particles at an early stage. Scrubbing combined with ultrasonic pre-treatment enhanced the liberation of surface-bound impurities and improved the efficiency of downstream separation. Calcination significantly reduced volatile species such as sulfates and chlorides, resulting in improved whiteness and enhanced mineral reactivity. Chemical leaching played a decisive role in final purification. Aqua regia treatment at 95 °C achieved approximately 96.8% removal of Fe₂O₃, leading to a substantial improvement in silica grade. Subsequent HF-HCl leaching enabled near-complete breakdown of refractory aluminosilicate phases, with Al₂O₃ removal exceeding 92%, thereby producing ultra-high-purity silica. Although the integrated physical-chemical flowsheet was highly effective in upgrading silica quality, it resulted in an overall reduction of approximately 20% in SiO₂ recovery relative to the original feed. This trade-off reflects the selective removal of impurity-bearing mineral phases, which is necessary to achieve ultra-high purity levels.

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Scrub typhus is a potentially fatal infectious disease caused by the obligate intracellular bacterium Orientia tsutsugamushi . While antibiotic treatment is generally effective, it requires extended treatment, and drug resistance and treatment failures have emerged. O. tsutsugamushi encodes a deubiquitylating enzyme, OtDUB, which interferes with host ubiquitin-dependent pathways. OtDUB cleaves ubiquitin from various substrates, but whether this activity can be selectively targeted by small molecules is unknown. Here we have screened a chemically diverse small-molecule library using a fluorescence-based deubiquitylation assay to identify potential inhibitors of OtDUB. Two compounds, gentisic acid and amiloride hydrochloride, inhibited OtDUB activity at low dosage, with little effect on the related Wolbachia CidB or yeast Ulp1 enzymes. Computational docking predicted the compounds engage regions near the OtDUB catalytic pocket, suggesting a competitive mode of inhibition; this was supported by enzyme kinetic analyses. Neither compound caused detectable cytotoxicity in mammalian cells. Amiloride hydrochloride treatment reduced both total cellular deubiquitylating activity and the O. tsutsugamushi bacterial load in infected cells. While the identified compounds are not optimized inhibitors, they establish that bacterial pathogen-encoded deubiquitylating enzymes can be targeted by small molecules. Overall, our results provide a framework for using selective inhibitors as tools to study DUB function in genetically intractable intracellular bacteria and as potential treatments for scrub typhus.

Accurate wetland mapping is essential for monitoring ecosystem health and guiding conservation planning efforts. This study evaluated the effectiveness of deep learning architectures and high-resolution National Agriculture Imagery Program (NAIP) imagery for classifying diverse vegetation cover across five public land holdings in three states with significant seasonally flooded, emergent wetland habitats in the southeastern United States. We built and tested deep learning models in ArcGIS Pro using three architectural encoders (U-Net, PSPNet, and DeepLabV3) with different input combinations, including NAIP natural color (RGB), Near-Infrared bands (NIR), Normalized Difference Vegetation Index (NDVI), Digital Elevation Models (DEM), and Texture metrics derived from NAIP imagery using Gray Level Cooccurrence Matrix (GLCM) features, resulting in eight (8) input combinations and three (3) rotational augmentations (0°, 30°, and 60°). Results revealed differences in classification accuracy ranging from 71 to 90% across sites, with Kappa scores from 0.54 to 0.81. We also found differences in model performance under alternating conditions and in the impact of combinations on performance metrics and computational time. Classification of the forested and scrub-shrub classes was consistent with F1 scores above 0.90 at several sites. However, distinguishing open water, aquatic beds, and emergent marshes became more difficult for some model-training architectures. At Mingo National Wildlife Refuge, clear spectral signatures and high-quality training data yielded the highest accuracy, thereby improving classification performance. In contrast, classification accuracy was lower for Red Slough and Choctaw East Wildlife Management Areas. Across all scenarios, the U-Net and DeepLabV3 models consistently outperformed the PSPNet model, achieving mean F1 scores close to 0.91, whereas PSPNet underperformed, with several configurations falling below 0.80. These results explain that while deep learning methods are effective and adaptable across different wetland types, their success depends on the ecological diversity of these environments. Overall, this research highlights the potential of deep learning for wetland monitoring and its crucial role in supporting conservation and management efforts.

Whole-genome sequencing is proving to be highly informative about the past demography of free-living populations, and in the context of endangered species, it can provide a quantification of the genetic risk posed by reduced genetic diversity and inbreeding. Prior to 1920, the Florida scrub-jay (Aphelocoma coerulescens) was more numerous across peninsular Florida, but with the expansion of agriculture and human habitation, its population has declined by 95%, resulting in fragmentation into semi-isolated subpopulations. By sequencing 241 individuals sampled from five different regions and across two time points, this study quantifies a greater loss of genetic diversity and greater levels of inbreeding in smaller and more isolated subpopulations. Consistent with population genetics theory, a reduction in population size results in a dramatic loss of rare alleles, skewing the site frequency spectrum far from the expected equilibrium. Increased inbreeding in the smaller, more remote subpopulations is especially evident in the increased size and number of runs of homozygosity. The Florida scrub-jay displays limited dispersal, and habitat fragmentation has greatly reduced the magnitude of gene flow in the past 30 years, resulting in further decline of genetic diversity, especially in the peripheral populations. Analysis of these data is informative in guiding conservation efforts to retain genetic diversity and minimize the consequences of inbreeding in the Florida scrub-jay.

Intraoperative C-arm imaging is essential in orthopaedic and trauma surgery, particularly with the increasing use of minimally invasive techniques. Conventional repositioning is performed by non-scrubbed staff, while robotic imaging systems are mainly limited to hybrid operating rooms. This study presents the first clinical experience with a fully motorised, self-driving mobile 3D-C-arm. In this prospective, single-center study, 300 procedures were analyzed using device log data and perioperative parameters to assess imaging workflow and system performance. Of 300 procedures, 279 were included. Mean procedure time was 104.7 ± 57.4 min, with a C-arm operation time of 31.9 ± 29.1 min, corresponding to a C-arm operation ratio (COR) of 35.2%. Automated positioning accounted for 30.7% of movements, and 37.9% of images were acquired from stored positions. The system enabled sterile-field control and demonstrated potential to support intraoperative imaging workflow. Further comparative studies are required to evaluate its clinical impact.

Leptospirosis is a zoonotic disease prevalent in low and middle-income countries and tropical regions. It can be mimicked by endemic infectious diseases that present simultaneously, like dengue, malaria, melioidosis, or scrub typhus. However, there is a dearth of documented evidence about the frequency of coinfection, as well as related symptoms and mortality. The study aims to determine the frequency and geographical extent of coinfections in leptospirosis patients in India. We systematically searched databases, including Scopus, PubMed, Embase, and Web of Science, for articles published between January 2010 and August 2024 assessing coinfections in leptospirosis patients in India. The eligible articles were evaluated for full text data extraction using a standardized methodology. Of 804 articles screened 39 studies were included; overall these reported on 1565 leptospirosis cases with 236 (15.1%) cases with coinfections. Of these, seven observational studies assessed coinfections in 1318 leptospirosis-positive cases, identifying 113 (8.6%) coinfections: most commonly dengue (79; 69.9%), followed by hepatitis E (15; 13.2%), typhoid (9; 7.9%), and malaria (9; 7.9%). The most coinfection cases were reported from New Delhi (29.6%), followed by Kerala (21.6%), Karnataka (14.4%), Uttar Pradesh (12.2%), and Tamil Nadu (5.5%). Common symptoms included fever, vomiting, icterus, headache, thrombocytopenia, and transaminitis. Cases with coinfection had poorer outcomes with increased mortality compared to cases with isolated infection. In India, leptospirosis can present with co-infections such as dengue, hepatitis E, malaria, typhoid, and scrub typhus, complicating diagnosis due to overlapping clinical features. These coinfections are associated with unfavorable outcomes, including multi-organ dysfunction and increased mortality. Early diagnostic testing in febrile patients and timely, appropriate treatment are essential to improve patient outcomes. Clinical trial number: not applicable. Review protocol registration (Open Science Framework): DOI https://doi.org/10.17605/OSF.IO/AS9W5.

Vector-borne diseases (VBDs) impose a significant burden in China, with complex spatiotemporal patterns driven by heterogeneous and nonlinear climatic influences, complicating the identification of causal drivers. To fill this gap, we conducted a nationwide study across 365 city-level regions in mainland China, covering nearly 0.8 million cases of eight types of VBDs, with a time span of 18 years. An empirical dynamic modeling framework was applied to reconstruct system dynamics directly from time-series data, and identify causal climatic drivers for eight VBDs via convergent cross-mapping. Their nonlinear effects and interactions were quantified via scenario exploration across diverse climate zones. Our results reveal distinct, disease-specific climatic drivers: dengue, Japanese encephalitis, hemorrhagic fever with renal syndrome (HFRS), and leptospirosis are primarily influenced by relative humidity and sunshine-hour; scrub typhus is sensitive to temperature, humidity, and sunshine; severe fever with thrombocytopenia syndrome (SFTS) and tick-borne encephalitis are mainly driven by sunshine-hour. Forecast improvement confirms marked north-south heterogeneity - sunshine alone improves dengue forecasts nationally, but both humidity and sunshine contribute in northern cities, whereas marginal effects diminish in persistently humid southern regions. Scenario exploration further uncovers nonlinear interactions and threshold effects: the positive effect of relative humidity on scrub typhus weakens above 25°C, and sunshine-hour shift from promoting to suppressing SFTS incidence across gradients. The climatic driving mechanisms of multiple VBDs in China were disease-specific, regionally heterogeneous, and state-dependent. The forecast-improvement effects of the identified climatic drivers differed between northern and southern regions, and their impacts changed nonlinearly across background climatic states.

Conservation actions such as habitat restoration and translocation require spatially informed, quantitative decision-making. We modeled habitat suitability for nine priority landbird species of the Channel Islands to understand habitat preferences, inform conservation planning and assess future climate impacts. In the absence of recent airborne lidar, we derived vegetation structure from spaceborne lidar, radar and optical remote sensing data combined with downscaled climate observations to train machine-learning models. Models performed well (average AUC = 0.84) under spatial cross-validation. Incorporating citizen science data improved Boyce Index performance for eight of the nine species, though AUC declined for 6 out of 9 species, likely reflecting spatial bias in opportunistic records. A radar-derived vegetation index was consistently influential, while maximum temperature strongly affected species restricted to Santa Cruz Island. Most species are projected to lose suitable habitat under two near-future (2040-2069) climate scenarios, except Grasshopper sparrow, Island scrub jay and Rufous-crowned sparrow. We identified climate change refugia to inform spatial conservation priorities.

Clinical audit is an effective tool to improve patient care, which in turn has an impact on clinical outcome.Patients who present with fever usually are generally admitted based on competency-based criteria, where admission is determined on the severity of the illness assessed through clinical and laboratory findings. Bacterial infections were linked to high sepsis-related mortality. This highlights the importance of comprehensive and standardized admission clerking and physical assessment for patients presenting with fever. This audit aimed to evaluate the documentation of history, physical examination, and investigations in febrile patients by postgraduate trainees at a tertiary care hospital in South India. A prospective audit of 50 case records of patients admitted with fever was conducted over a four-week period in the medical wards. A predefined pro forma was used to review key domains, including documentation of admission details, clinical history, physical examination (vital signs and systemic examination), and relevant investigations. Findings were measured against departmental standards and analyzed to identify areas for improvement. Documentation completeness by the post graduate trainee's completeness varied across domains. Admission details were documented in 86% of cases, and comprehensive history was recorded in 72%. While vital signs were documented well (>90%), systemic examination showed significant variability (CVS: 80% vs CNS: 66%). Documentation of specific investigations was inconsistent: dengue serology (62%), WIDAL test or blood culture (54%), and Scrub typhus serology (38%). The clinical audit revealed notable variability in admission documentation by postgraduate trainees, particularly in the systemic examination and investigation domains. The findings highlight the need for targeted educational interventions and reinforcement of documentation protocols to enhance patient assessment and care, particularly in febrile patients, where inadequate management can lead to high case fatality.

Orientia tsutsugamushi is a mite-transmitted obligate intracellular bacterium that causes the potentially deadly zoonosis, scrub typhus. The absence of genetic tools for Orientia has limited studies of the microbe-host interactions that underlie scrub typhus. To address this gap, we developed a protocol for transforming and achieving allelic exchange in O. tsutsugamushi str. Ikeda. From evaluating multiple cell lines and antibiotics, we found that contact-inhibited EA.hy926 human endothelial-like cells best supported Orientia replication, and that chloramphenicol was an effective selection marker. We engineered a homologous recombination cassette encoding a codon-modified version of the O. tsutsugamushi ank13 gene (OTT_RS04140) (CMank13) and its promoter alongside genes for mScarlet-I and chloramphenicol acetyltransferase under control of the O. tsutsugamushi tsa22-up and tsa56-down promoters, respectively. A PCR product encompassing the cassette and chromosomal flanking regions was transformed into O. tsutsugamushi via electroporation or CaCl2, the latter of which better preserved bacterial and host cell viability. EA.hy926 cells inoculated with transformed O. tsutsugamushi were grown in glass-bottom plates in the presence of chloramphenicol and imaged by live-cell microscopy to identify cultures containing mScarlet-I-positive bacteria, which could be maintained in perpetuity. Chromosomal integration of the CMank13 cassette and loss of wild-type ank13 were verified by PCR and nanopore sequencing. This report establishes platforms for genetically manipulating O. tsutsugamushi and building additional genetic tools to investigate this globally significant pathogen. Orientia tsutsugamushi causes scrub typhus, a globally emerging rickettsiosis that can have a high mortality rate and has been a known human disease since the fourth century. Of the genera of obligate intracellular bacterial pathogens that cause human disease, Orientia is the only one for which genetic tools have not been developed. This has limited understanding of O. tsutsugamushi-host dynamics that drive the bacterium's pathobiology and hinder the development of novel treatment or protection strategies against scrub typhus. Here, we successfully transformed and achieved allelic exchange in O. tsutsugamushi. Transgenic bacteria were selected via antibiotic resistance, validated by PCR and nanopore sequencing, and visualized by immunofluorescence and live-cell fluorescence imaging. Our report includes detailed descriptions of empirically determined host cell cultivation, multiplicity of infection, transformation, and selection conditions to provide a foundation on which other researchers can build. Overall, this work begins to establish a genetic toolbox for O. tsutsugamushi.