Open Research Europe

BackgroundMalaria control is increasingly challenged by the urban-adapted vector Anopheles stephensi, yet molecular and cellular tools for this species remain scarce, restricting functional genomic studies and the development of genetic control strategies. To help address this gap, we established a new embryo-derived Anopheles stephensi cell line. ResultsWe generated and characterised a novel embryo-derived Anopheles stephensi (SDA-500) cell line capable of sustained growth in vitro. Species identity was confirmed by mitochondrial COI barcoding, and karyotypic analysis revealed a diploid chromosome complement with the presence of a Y chromosome, confirming that at least some cells are of male origin. Transfection conditions were optimized, with TransIT-PRO showing higher efficiency than Lipofectamine-based reagents. Using a dual-luciferase reporter assay, of several promoters tested the Anopheles gambiae polyubiquitin promoter exhibited the strongest and most consistent transcriptional activity in SDA-500 cells. ConclusionsThe SDA-500 cell line provides a stable and genetically validated in vitro platform that supports efficient transgene expression. This resource provides a useful system for functional genomics and molecular manipulation in Anopheles stephensi and is expected to facilitate studies of mosquito biology and contribute to the development of novel malaria control strategies.

Background. Correct and accurate deconvolution of SARS-CoV-2 lineages from wastewater sequencing data is a challenging task, given the intricacy of wastewater amplicon-sequencing data and the ever-growing complexity of the lineage classification. Existing benchmarking studies made use of artificial spike-in compositions, thereby falling short of reflecting the prevailing complexity of wastewater samples. Results. We present a modular, expandable, simulation-based benchmarking framework as a reproducible Snakemake workflow, named Laemple, to evaluate the performance of virus lineage deconvolution tools. Using in silico simulated data sets of varying complexity and sequencing quality, we demonstrate its utility by evaluating seven publicly available tools, based on their precision, sensitivity, and reproducibility. Freyja showed robust sensitivity and consistent performance across diverse data set complexities, alongside user-friendly installation and documentation, while VaQuERo demonstrated the highest precision. Conclusions. Our results reveal substantial variation in tool performance across conditions, emphasizing the need to benchmark with diverse and complex scenarios. This framework enables informed tool selection for researchers and public health agencies and allows developers to stress-test their tools during development and maintenance, i.e., updating the lineage definition for newly emerging virus lineages. To this end, Laemple was designed in a modular fashion for customization and future expansion to support ongoing software development across all stages of the application life-cycle management.

Occupational ocular injuries are a preventable yet neglected public health problem, particularly in low- and middle-income countries. Maintenance workers are exposed to diverse ocular hazards daily, yet compliance with protective measures is consistently poor. A descriptive cross-sectional study was conducted among 85 maintenance workers at the Maintenance and Essential Services Organization (MESO) of Kwame Nkrumah University of Science and Technology (KNUST), Ghana, recruited through stratified convenience sampling across seven occupational sections. A structured questionnaire assessed knowledge of ocular hazards and protective equipment, attitudes toward ocular safety, and safety practices. Data were analyzed using IBM SPSS version 26 (IBM Corp., Armonk, NY, USA); chi-square and Fishers exact tests assessed associations (p < 0.05). Participants were predominantly male (84/85, 98.8%), with a mean age of 44.5 {+/-} 10.4 years. Overall knowledge was good (mean 9.40 {+/-} 1.59 out of 11), but attitude and practice scores were average (2.78 {+/-} 0.92 and 3.27 {+/-} 0.93, respectively). Most workers correctly identified goggles and face shields as protective, but only about half recognized that ordinary sunglasses and spectacles offer inadequate protection. Although 97.6% (83/85) recognized the need for ocular protection, only 7.1% (6/85) reported consistent protective eyewear use, and fewer than half (45.9%, 39/85) had received formal ocular safety training. Routine general protective equipment use was significantly associated with ocular protection use (Fishers exact test, p = 0.011). Sand and dust particles were the leading causes of injury and only 25% (5/20) of injured workers sought formal care. Workers demonstrated good knowledge but poor attitudes and practices toward ocular safety, suggesting that knowledge alone does not translate into protective behaviour even within a relatively well-resourced institutional setting. Findings suggest that limited access to task-appropriate protective eyewear may represent an important institutional barrier. Institutional PPE supply and section-specific safety training are essential to bridge this knowledge-practice gap.

Uniform, reproducible blood smears are critical for reliable hematological evaluation. Manual smear preparation, however, is user-dependent and introduces variability that limits quantitative microscopy. Here we developed BUsmear, a low-cost, 3D-printed, motorized blood smear device that prepares two smears simultaneously from a printed stage and a micro-motor drive with tunable linear velocity, controlled through a joystick-operated driver module. By spreading two slides in parallel with fully repeatable slide-to-slide motion, the device doubles throughput while eliminating operator-dependent motion artifacts, and can be fabricated on any benchtop 3D printer in under one day. To validate smear quality, we analyzed blood films from three donors together with a manual smear prepared by an expert from the blood of one of the same donors, giving a matched device-versus-manual pair. Automated Cellpose segmentation of 14,046 red blood cells across 12 bright-field fields showed that cell diameter was preserved and closely matched the expert smear (6.7-7.8 um across groups, within the 6.2-8.2 um human reference range; 6.7 vs 6.9 um in the matched pair), and that all films formed non-aggregated monolayers (Clark-Evans index of aggregation 1.04-1.24). Critically, red blood cells in the device films were markedly more circular than in the expert manual smear (mean eccentricity 0.405 vs 0.502; 0.443 vs 0.502 in the matched pair), with complete separation between the two methods at the level of whole fields of view. Because eccentricity reports smear-induced cell distortion, this indicates that a constant, mechanically controlled spreading velocity preserves red blood cell morphology better than skilled manual technique. BUsmear offers an accessible route to standardized smear geometry for quantitative analysis, including AI-based morphometry, and its low cost may be particularly advantageous in low-income countries with a high prevalence of malaria.

Malaria transmission depends on the survival and mosquito infectivity of mature Plasmodium falciparum gametocytes. Despite malaria febrile episodes reaching 39 to 41.5{degrees}C in infected humans, the impact of febrile temperatures of varying durations on gametocyte viability and transmissibility remains undefined. Here we quantify the effects of exposing mature stage V gametocytes in vitro to febrile temperatures (39{degrees}C, 40{degrees}C, and 41.5{degrees}C) for varying durations (3 to 12 hours). To assess gametocyte morphology, functionality, and mosquito infectivity, we used light microscopy of Giemsa-stained thin blood smears, exflagellation assays, and standard membrane feeding assays (SMFAs). Following exposure to 39{degrees}C, gametocytes retained normal morphology, exflagellation capacity, and mosquito infectivity across all exposure durations. At 40{degrees}C, gametocytes remained morphologically intact and capable of exflagellation but exhibited a time-dependent reduction in mosquito infection, both in prevalence and oocyst intensity, following prolonged exposure. In contrast, exposure of mature gametocytes to 41.5{degrees}C, even for the minimum duration tested of 3 hours, resulted in complete loss of normal morphology, exflagellation capacity and ability to infect mosquitoes. These findings reveal that mosquito infectivity of mature gametocytes is affected both by the magnitude of the febrile temperature and the duration of exposure. This provides insight into how host fever dynamics may influence parasite transmission.

Background Onchocerciasis (river blindness), caused by the filarial nematode Onchocerca volvulus and transmitted by Simulium blackflies, remains a public health challenge in sub-Saharan Africa. Following the transition from vector control to large-scale ivermectin distribution, microfilariae (mf) prevalence has declined substantially across Ghana. The World Health Organization targets onchocerciasis elimination, defined as sustained mf prevalence below 1%. This requires identifying areas of persistent or resurgent transmission to guide targeted mapping surveys and interventions. Methods A Bayesian spatiotemporal, multilevel, generalized additive model with a zero-inflated beta-binomial outcome was fitted to mf prevalence data from 1353 surveys across 671 villages and environmental and socio-economic predictors. Temporal prevalence trends for ecozones within operational transmission zones (OTZ) were modelled using a 2D spatial smooth to account for residual spatial autocorrelation. Temporal autocorrelation in prevalence was accounted for using a random effect for village nested within ecozone. Results Runoff probability and surface wetness were positively associated with prevalence, consistent with Simulium breeding in fast-flowing water. Despite high initial mf prevalence, by 2015 the median prevalence across all OTZs in Ghana was estimated to be between 0.43 and 1.97%. The probability that prevalence was <1% in 2015 ranged from 0.15 to 0.87. Assuming testing rates and mass drug administration have remained consistent (twice a year and >80% coverage), linear extrapolation predicted that the three OTZs have some probability of exceeding the elimination threshold by 2030, while two were predicted to have near-zero prevalence. The Tano-Ankobra OTZ, which includes both savanna and forest agro-ecological zones, was a persistent hotspot. Conclusions This first comprehensive spatio-temporal analysis of onchocerciasis in Ghana reflects the well-documented challenge of onchocerciasis control in forested riverine environments. Persistent transmission foci in eastern and southern Ghana, particularly in forest zones with sustained vector habitat, warrant targeted investigation and intensified intervention to meet WHO elimination thresholds.

Disinfection with bleach is recommended for surface cleaning in healthcare facilities, but bleach degrades over time, reducing disinfection effectiveness. We evaluated whether local bleach produced by hospitals using novel technology had adequate shelf life to justify use. Two hospitals (A and B) each produced and stored 0.5% bleach in two 20-liter plastic containers; bleach in one container was alkalinized for increased stability to pH 12; bleach in the other container was not stabilized and, for comparison, a third container of commercial bleach was tested. We tested three bleach samples from each of the containers produced by hospitals A and B, and commercial bleach for free available chlorine (FAC) using N,N-diethyl-p-phenylenediamine at irregular intervals up to 216 days. We compared the expected percent decrease in FAC per 7 days by bleach type using log-normal regression. Stabilized bleach decreased by 1% and 2% in hospitals A and B, respectively, every 7 days while non-stabilized bleach decreased by 21% and 12%, respectively (all p<0.001); commercial bleach decreased by <1% per 7 days. Bleach production proved feasible in hospitals, and both stabilized and commercial bleach maintained adequate concentration for disinfection; non-stabilized bleach maintains a useful concentration for not longer than three weeks after production.

High-speed atomic force microscopy (HS-AFM) experiments provide unique insights into nanoscale biological processes by directly visualizing the surface topography dynamics of biomolecules, such as proteins, over extended periods of time and generating large imaging data sets. However, the lack of computational methods for automated data analysis has thus far limited the full utilization of these data for the quantitative understanding of single-molecule dynamic. Here, we present the AFM Analysis Arena (A3) within BioAFMviewer as a software platform for automated, high-throughput quantitative analysis of AFM topographic imaging data within a user-friendly interactive graphical interface, implementing a convenient workflow from raw HS-AFM (AFM) imaging to statistical analysis. In several applications to HS-AFM imaging data, we demonstrate methods of automatized molecule detection and dynamic tracking to highlight the available capacities, aiming to transform HS-AFM measurements from observations towards an analytical tool in nanoscale biology.

Population genomic studies have revealed strong regional structure in Plasmodium knowlesi across Southeast Asia. To facilitate routine genomic surveillance, we developed a targeted amplicon sequencing panel that preserves the large- and small-scale population genetic structure. We screened published genomes for short, highly polymorphic targets and a 30-SNP barcode. We validated multiplexed amplification and sequencing using a portable Oxford Nanopore Technologies MinION sequencer on 28 clinical infections from four southern Thai provinces collected between 2018-2023. Twenty-seven loci yielded sequence data and 23 infections with data from at least eight loci were retained for analysis. Across the 23 Thai infections and 104 published Malaysian genomes, 586 diverse SNPs were observed. The amplicon panel and 30-SNP barcode retained the major population structure observed in genome-wide data, including a clear distinction between Bornean and Peninsular Malaysian infections and two clusters in Borneo. Across newly sequenced Thai infections, 25 loci were diverse and showed substantial diversity (mean expected heterozygosity = 0.69). All infections were effectively monoclonal. Pairwise identity-by-descent revealed substantial recent relatedness, with 46 of 253 pairs at or above 0.5. Two Thai genetic groups (n = 12 and n = 11) were observed, mirroring previously observed structure in peninsular Malaysia. Groups were not significantly associated with province or collection year. In conclusion, targeted sequencing can reveal geographically informative population structure across scales and thus support surveillance.

The photic sneeze reflex (PSR), in which exposure to bright light triggers sneezing, is widely recognised but inconsistently defined and measured. We conducted the first systematic review and meta-analysis of PSR prevalence to synthesise the epidemiological evidence, assess methodological quality, and identify priorities for future research. We included 18 articles comprising 31 study groups and extracted prevalence estimates, study characteristics, ascertainment methods, and epidemiological information. Fifteen eligible study groups classified as healthy were included in the primary meta-analysis. The pooled prevalence was 22% (95% CI, 15%-29%), with extreme between-study heterogeneity (I2 = 99.3%). Reported prevalence estimates and associations with participant characteristics varied widely, and nearly all studies were judged to be at high risk of bias. The pooled estimate should therefore be interpreted as a descriptive summary of the available evidence rather than a precise estimate of population prevalence. Future studies require a standardised operational definition, representative sampling, transparent reporting, and reproducible methods for assessing light-triggered sneezing.

Previously, we have demonstrated that certain ferric salts common in Archean waters, such as iron trichloride and ferric ammonium citrate, can protect liposomes from destruction by short-wavelength UVC light. In this study, we investigate the propagation of 254 nm UV radiation through aqueous FeCl3 solutions and its interactions with liposomes. We then consider these findings in the context of early Earth UV environment, discuss their implications for our hypothesis of the Darwinian evolution of liposomes, and integrate them with our previous experimental results.

Cannabis sativa L. cultivation in tropical regions or under indoor conditions requires supplemental lighting to maintain plant yields. Night-break (NB) photoperiods offer a cost-effective alternative to conventional day extension, although genotypic responses have not been clearly described. This study evaluated the effects of continuous and pulsed NB treatments on the growth, phenology, physiology, productivity, and cannabinoid accumulation of three Cannabis sativa genotypes representing two chemotypes. Plants were exposed to a commercial 18-h photoperiod (T1), a continuous 1-h NB (T2), or four 15-min NB pulses (T3) with equal total duration. Continuous NB produced responses comparable to the commercial photoperiod. In contrast, pulsed NB accelerated phenological progression by two to three weeks and reduced final plant height by 15-28%, depending on genotype, without impairing photosynthetic performance or decreasing floral dry biomass. Harvest index increased under both NB treatments, indicating enhanced biomass allocation to reproductive structures, while cannabinoid accumulation remained primarily genotype-dependent with only minor treatment-specific modulation. These findings demonstrate that NB efficacy depends not only on the duration of nocturnal light but also on its temporal distribution, highlighting temporal organization of nocturnal light as a key regulator of photoperiodic responses and a promising strategy for optimizing cannabis cultivation.

Background In Nigeria, the country with the greatest global malaria burden, funding constraints increasingly require insecticide-treated net (ITN) reprioritization to target those at highest risk. Large language models (LLM)- assisted decision-support tools may facilitate risk-informed ITN planning by supporting malaria programme officers in navigating analyses, interpreting outputs, and translating evidence into operational decisions. We developed ChatMRPT, an LLM-assisted ITN allocation planning tool based on user requirements, and analyzed post-interaction feedback, examined user digital competencies and workplace resources, and identified institutionalization pathways for LLM-assisted intervention planning. Methods A two-phase mixed-methods study began with software requirements gathering workshops (using a prototype) involving representatives from the National Malaria Elimination Programme (NMEP), State Malaria Elimination Programmes (SMEPs), and implementing partners. Phase 2two evaluated ChatMRPT through surveys, guided exercises, and focus group discussions with 34 SMEP officers from 28 Nigerian states. Quantitative data were analyzed using descriptive statistics and profile-based comparisons, while qualitative data were analyzed using reflexive thematic analysis to synthesize user experiences of ChatMRPT and identify institutionalization pathways. Findings Fifty-eight percent (19/33) of participants demonstrated both higher digital competency and adequate workplace resources; the remainder exhibited limitations in one or both domains ([4/33] higher competency/constrained resources; [6/33] higher resources/lower competency). Participants with higher digital competency but constrained workplace resources reported user experiences comparable to those with higher competency and adequate resources, whereas workplace resources alone did not appear to compensate for lower digital competency. Key software requirements included contextual guidance for malaria risk interpretation, operational decision support, and embedded analytical support. Following iterative incorporation of these requirements, ChatMRPT was positively evaluated across participant profiles. Participants viewed institutionalization as dependent on integration into routine malaria planning and adaptability to evolving programme priorities. Interpretation Many malaria programme officers may already have the foundational competency for LLM-assisted decision support. However, there is room to further strengthen digital competencies while facilitating access to basic workplace resources such as stable internet. Institutionalization of LLM tools may depend on addressing these capacity and infrastructural constraints alongside designing explainable, integrated, and flexible systems. Future research should evaluate long-term integration, sustainability, and effectiveness in routine malaria planning. Funding This work was funded by the Bill and Melinda Gates Foundation (INV-036449) and the Center for Health Outcomes and Informatics Research (CHOIR), Loyola University Chicago. The funders had no role in the study design, data analysis, interpretation of findings, or preparation of the manuscript.

The Arabian killifish (Aphaniops dispar) is an excellent model for studying human diseases such as fungal infections and cancer progression. The embryos possess a transparent chorion for live imaging and display an extended period of independent feeding (13 days post-fertilisation). They also exhibit broad thermal tolerance which enables live imaging at physiologically relevant human temperatures. However, omics resources for this species remain limited and restricts its use in genomics studies. We generated a 1.45 GB high-quality reference assembly with 24 chromosome-level scaffolds and N50 score of 60.64 Mb by combining long-read, long-range, and short-read sequencing. The A. dispar genome exhibits a high level of homozygosity (95.6%) with BUSCO gene completeness scores of 99.2% and 97.5% for the Actinopterygii and Cyprinodontiformes lineages, respectively. The genome is highly repetitive (58.7%) and the majority of these are DNA transposons. The mitochondrial genome is 81.55% similar to that of Aphanius iberus (the Spanish toothcarp), with minor structural modifications. A. dispar exhibits highly conserved synteny with Fundulus heteroclitus within the Cyprinodontoidei clade. Together, these findings provide a valuable genomic resource for comparative genomics within the teleost community.

Menstrual health remains a neglected public health challenge in India, where persistent stigma, incomplete education, inadequate sanitation, and limited access to menstrual health resources can adversely affect health and social participation. Institution-based educational programs represent an underexplored opportunity to improve menstrual health literacy while promoting more inclusive discussions surrounding menstruation. Bridging this gap, an exploratory, prospective educational evaluation was conducted at a hospital in Coimbatore, Tamil Nadu and an engineering college in Wayanad, Kerala. Anonymous pre- and post-course surveys assessed menstrual health knowledge, attitudes, practices, perceived barriers, and perspectives on emerging digital health technologies. Quantitative responses were summarized descriptively, and open-ended responses were reviewed to identify recurring perspectives that contextualized the quantitative findings. The standardized educational program integrated evidence-based menstrual health education with discussions of bioengineering and wearable technologies. Eighty-three participants completed the pre-course survey, 68 attended the educational program, and 20 completed the post-course survey. Because responses could not be linked at the individual level, the survey cohorts were compared descriptively. At baseline, 80 of 82 respondents (97.6%) recognized menstruation as a biological process, yet only 38 of 82 (46.3%) indicated that it was not solely a woman's issue. The mean agreement that menstruation remained taboo was 7.44 of 10 (SD 1.99; n=78), and 51 of 77 respondents (66.2%) reported that they or others avoided activities during menstruation. Painful periods were identified as a barrier by 56 of 67 respondents (83.6%), and 18 of 67 (26.9%) considered institutional or community toilets insufficiently clean and private for menstrual management. Open-ended responses illustrated how pain, fear of leakage, stigma, and inadequate facilities constrained participation in school, physical activity, and social life. These findings document a marked gap between basic biological recognition and the social normalization and practical support of menstruation in two South Indian institutions, and support further evaluation of gender-inclusive, context-responsive menstrual health education using paired assessments, stronger follow-up, comparison groups, and long-term outcomes.

Background Scientific mistrust contributes to lower participation and underrepresentation of Black Americans in genomics studies limiting understanding of how genomic variation and environmental exposures influence health disparities. Community-engaged research requires rebuilding scientific trust; however, there is a need for practical models that operationalize guidance for researchers without community-engaged research training. Therefore, we developed an educational intervention for genomics researchers initiating partnerships with Black American communities. Our intervention creates a bidirectional teaching environment that allows potential community and academic partners to discuss areas of expertise for each stakeholder, partnership perspectives and needs, while exploring modules related to genomics research and community-academic partnership. We report a novel and structured approach for prospective academic and community partners to mutually orient one another and assess partnership practicability. Methods Ten participants, recruited through established community channels, attended a four-hour workshop containing nine interactive modules about scientific mistrust, research safeguards, community-engaged research, genomics, and research for community-defined goals. We also experimented with humor to enhance engagement and trust. We used mixed-methods, single-arm research design and assessed feasibility through recruitment success and retention. Using inductive rapid thematic analyses, we assessed participant responses to eight workshop prompts. Preliminary quantitative data, used for descriptive purposes due to low sample size, were analyzed from pre- and post-Likert scale surveys assessing associations between the intervention and four domains, including scientific trust. Results All 10 enrolled participants completed the study, meeting a priori criteria of 100% for recruitment and show rate. Engagement was strongest during early workshop modules and declined in later modules. Survey data completeness was limited by missing responses. Survey instrument design limitations were identified for modification in future studies. Participants articulated expectations for partnership that aligned with community-based participatory research principles. Conclusions The intervention is feasible to deliver as a bidirectional educational experience in partnership with a community organization. The strongest implementation refinements needed were workshop duration, module prioritization, and survey instrument design, particularly the trust domain. A community advisory board is co-developing modules and refining the intervention to evaluate in a pilot version of the study with a larger sample.

Life evolved under broad-spectrum sunlight (300-3500 nm), but most modern light-emitting diodes (LEDs) emit within a restricted visible range (350-650 nm) leaving lighting environments that are short-wavelength dominant. We ask if chronic 450nm exposure dominant in LEDs alters metabolism and organ integrity in ageing mice environmentally exposed at 13 mW/cm{superscript 2} for 5 h/day over 40 weeks, at an intensity within the range of human environmental exposures. Mice exhibited progressive weight gain despite reduced food intake, with altered glucose tolerance and insulin sensitivity over time. White adipose tissue mass increased, while brown adipose tissue weight and UCP1 protein levels remained unchanged. Relative organ weights (heart, liver, kidney, testis) were each reduced by ~10%, with histopathology revealing fat deposition, hepatocellular degeneration, renal tubular damage, steatosis, and impaired testicular architecture. These findings demonstrate that chronic 450 nm light exposure is associated with metabolic alterations and multiorgan pathology in mice.

Life evolved under broad spectrum sunlight (300-2500nm), but in the modern built environment under standard Light Emitting Diode (LED) illumination is restricted to around 400-650nm. Different wavelengths influence metabolism via mitochondria. 670-900nm support mitochondrial function while those in the Soret band (420-450nm) undermine mitochondrial ability. Under LED lighting, metabolism is challenged with reduced mitochondria ability, and hence, reduced demand for blood sugars that are required to produce adenosine triphosphates (ATP). As longer wavelengths enhance mitochondrial performance, they increase demand for blood sugars, reducing serum glucose. Here we change the lighting in windowless offices providing either standard LED desk lamps or those supplemented with 850nm in real-world environments. This was for two hours every morning over three days. We then assessed blood sugars with two independent methods, an oral glucose tolerance test and continuous glucose monitoring. Subjects exposed to standard LEDs had significantly increased blood sugars, while those exposed to lamps with added 850nm had significantly reduced blood sugars. These real-world data highlight the importance of light in controlling metabolism. They raise issues concerning public health in interior environments where we now spend >90% of our lives.

Salmonella is a pathogen linked to foodborne outbreaks, including low-moisture foods. Its ability to resist desiccation can contribute to the formation of dry surface biofilms (DSB). This study evaluated the impact of 3 DSB formation protocols (P1=48-h hydrated phase/48-h dry phase, P2=24-h/120-h and P3=8-h/48-h) on the resistance of Salmonella DSB to 70% alcohol, a commercial product (based on 0.015% quaternary ammonium and 25% isopropyl alcohol), gaseous ozone (45 ppm), hot air (90 {degrees}C) and UV-C light (254 nm). The type of DSB protocol impacted the efficacy of the sanitizers (p < 0.05). The biofilm with the shortest hydration phase showed the greatest susceptibility; three out of the five sanitizers evaluated (70% alcohol, commercial product, and UV-C) promoted significant reductions in P3, with counts below the detection limit (0.8 log CFU/cm{superscript 2}) after 5 to 15 min exposure. Regarding protocols P1 and P2, in general, the best performance was from UV-C, especially against DSB on polypropylene, where it achieved reductions of 1.3 log CFU/cm{superscript 2} for P1 and 2.9 log CFU/cm{superscript 2} for P2 after 15 to 30 min of exposure. In contrast, hot air and ozone showed less effectiveness, with reductions [&le;]1.2 log CFU/cm{superscript 2}. In most scenarios, confocal microscopy images corroborated the plate count results (log CFU/cm{superscript 2}). In summary, our data indicates limited action of dry sanitizers on Salmonella DSB, requiring validation and optimization of sanitization processes to ensure the microbiological safety of low-moisture products.

Background: Hand hygiene is a key public health intervention for preventing infectious diseases, including respiratory and gastrointestinal infections. Although the COVID-19 pandemic improved awareness and compliance, sustained adherence remains a challenge, with evidence showing a decline in practice after outbreaks, particularly among young adults in high-contact settings such as universities. Therefore, this study aimed to assess the knowledge, attitudes, and practices of handwashing hygiene among university students at KCMC University, Moshi, Tanzania, in the post-COVID-19 era. Methods: A cross-sectional study was conducted among 380 students at KCMC University. Data were collected using a pretested structured questionnaire, which captured information on socio-demographic characteristics, knowledge, attitudes, and practices regarding handwashing hygiene. Data were analyzed using descriptive and inferential statistics. Associations between variables were assessed using logistic regression and multivariable analysis, and a p-value of <0.05 was considered statistically significant. Results: A total of 380 students participated in the study, with a mean age of 23.5 {+/-} 3.5 years. More than half 209 (55.0%) were aged 18-23 years, while 203 (53.4%) were female. Most participants had access to hand hygiene facilities 338 (88.9%). 338 (88.9%) participants had good knowledge of hand hygiene but attitudes were mostly negative 193 (50.8%), and practices were suboptimal, with 201 (52.9%) showing poor hand hygiene practices. Good hand hygiene knowledge was associated with access to hand hygiene facilities (AOR=2.84, 95% CI: 1.25-6.42, p=0.012) and formal training on hand hygiene (AOR=3.48, 95% CI: 1.24-9.76, p=0.018). Positive attitudes were associated with being in the fourth year or above (AOR=2.12, 95% CI: 1.16-3.89, p=0.015). Good hand hygiene practices were associated with access to hand hygiene facilities (AOR=2.67, 95% CI: 1.18-6.01, p=0.018) and formal training on hand hygiene (AOR=2.98, 95% CI: 1.19-7.48, p=0.020). Conclusion: Although knowledge and awareness of handwashing hygiene were generally high among university students, inconsistencies between knowledge and actual practices were observed in the post-COVID-19 era. Strengthening health education programs and ensuring continuous access to handwashing facilities may improve adherence to proper hand hygiene practices among university students.