Parasites & Vectors

BackgroundAttractive Targeted Sugar Baits (ATSBs) are a promising, environmentally friendly approach for mosquito control, but the direct field application, scalability and long-term effectiveness of ATSBs across diverse mosquito species remain significant challenges. Methodology/Principal FindingsWe assessed the efficacy of a genetically engineered RNA interference (RNAi) yeast strain (Sh.463_56.10R) formulated in three sugar baits, soda (Coca-ColaTM), 10% sucrose, and a commercial mosquito bait (BaitStabTM), on Aedes, Anopheles and Culex mosquitoes. All RNAi yeast bait formulations induced significantly higher mean mortality (87-100%) compared to the control groups (0-9%; P<0.0001), but mosquitoes exhibited a higher feeding preference for RNAi yeast-soda baits, which induced mortality rates of 94-100% (P < 0.0001) recorded across all mosquito species. Additionally, to assess the competitiveness of the RNAi yeast-soda bait to other tropical sugar sources, semi-field choice assays were conducted in Trinidad, West Indies using competing flowering plants and fruits typically found in residential environments. The RNAi yeast-soda ATSB continued to perform well in the presence of competing floral and fruit sugar sources during both Aedes albopictus and Culex quiquefasciatus trials, though the presence of several fruits and flowers did reduce A. aegypti mortality, suggesting that further field testing will be necessary. The residual activity of the Sh.463_56.10R + soda formulation was retained for at least 14 months, with sustained 100% mortality in C. quinquefasciatus and 93-100% mortality in Aedes aegypti, Anopheles gambiae and Anopheles stephensi. The RNAi yeast-soda ATSB also performed well in semi-field studies performed with a prototype soda bottle feeder. Conclusions/SignificanceThis study demonstrates the potential of soda-baited RNAi yeast as a potent, long-lasting, and scalable platform for ATSB-based mosquito control as a component of integrated vector management programs. Author SummaryMosquito-borne diseases continue to affect millions of people worldwide, and current mosquito control methods face challenges such as low public uptake, insecticide resistance and environmental concerns. Here we evaluated a new and environmentally friendly approach to mosquito control using ATSBs. We tested genetically engineered species-specific yeast producing RNAi molecules capable of killing mosquitoes that feed on it. We mixed the yeast with three different sugar baits, including soda (Coca-ColaTM), 10% sucrose, and the commercial mosquito bait BaitStabTM formulation, and evaluated how well they worked against different mosquitoes. The results showed that the RNAi yeast mixed with soda was the most effective, killing up to 100% of mosquitoes in laboratory and outdoor tests. The bait remained effective in the presence of many competing natural tropical fruit and floral sugar sources. Remarkably, the bait, which can be delivered in a soda bottle feeder, stayed active for at least 14 months under simulated field conditions. These findings suggest that soda-based RNAi yeast baits could provide a practical, long-lasting and scalable tool for mosquito control and may help strengthen future strategies to reduce mosquito-borne diseases.

Background: The lifespan of insecticide-treated nets (ITNs) varies widely across settings, reflecting both intrinsic product characteristics and external factors related to use, care, and environment. While the resistance to damage (RD) score captures intrinsic product durability, there is no standardized metric to quantify contextual risks. This study presents a proof of concept for the Risk Index (RI), a composite measure of site-level risk factors for ITN physical durability and survival. Methods: We conducted a secondary analysis of durability monitoring data from 44 sites across 15 countries in sub-Saharan Africa, covering 14 ITN products. The RI was calculated as a weighted composite of 12 indicators spanning net handling, net care attitudes, and use environment. Associations between RI and median ITN survival were assessed using weighted linear regression and multivariable mixed-effects models adjusting for RD score, with country included as a random effect. Results: RI scores ranged from 25.1 to 83.7 across study sites. In bivariable analysis, a 10-point decrease in RI was associated with a 4.0-month increase in median ITN survival (95% CI: 1.7-6.3; p=0.001). In multivariable analysis adjusting for RD, this association remained significant but attenuated to 2.2 months (95% CI: 0.1-4.2; p=0.037). Independently, a 10-point increase in RD score was associated with a 3.5-month increase in survival (95% CI: 1.3-5.7; p=0.001). No interaction was observed between RI and RD. Predicted survival differed by approximately one year between the lowest- and highest-risk settings. Conclusion: The RI provides a standardized measure of contextual risk factors affecting ITN lifespan, independent of ITN product type. When used alongside a product's RD score, the RI enables improved interpretation of expected site-level variation in net performance. This combined framework offers a practical basis for incorporating behavioural and environmental risk into vector control planning and for tailoring ITN strategies to local conditions.

Rhodnius prolixus is a primary insect vector of Trypanosoma cruzi, the causative agent of Chagas disease, a neglected parasitosis endemic in Latin American countries. It has been estimated that Chagas disease affects 7-8 million people worldwide and is responsible for approximately 1000 deaths per year. Genetic and molecular studies in this species remain challenging due to its life cycle and feeding habits, thus hindering the development of new strategies to control their populations and reduce the diffusion of Chagas disease. Recently, two stable cell lines - RPE/LULS53 and RPE/LULS57 - were derived from Rhodnius embryos, which represent promising new tools to investigate the genetics of this insect vector. Here, we describe their gene expression landscapes through transcriptomic approaches. We show that 8,968 expressed genes are shared between the two cell lines, whereas 391 and 1,088 genes are uniquely expressed in RPE/LULS53 and RPE/LULS57, respectively. Although key components of primary developmental, immune and redox signaling pathways are expressed in both cell lines, some genes such as Frizzled-10-a-like and catalase show marked differences in expression. Our results strongly suggest that RPE/LULS53 and RPE/LULS57 likely represent two different cell phenotypes. Consistent with this, gene ontology analysis reveals that RPE/LULS53 is enriched for animal organ morphogenesis and stress response, while RPE/LULS57 for DNA-directed RNA polymerase activity, among others. Despite these differences, both cell lines express comparable levels of transcripts from resident transposable elements, including the highly abundant Mariner and LINE/I elements, as well as horizontally transferred transposons. Our findings shed light on the nature of the RPE/LULS53 and RPE/LULS57 embryo-derived cell lines and provide valuable transcriptomic resources for future genetic and functional studies in Rhodnius and other triatomine insect vectors. Author summaryRhodnius prolixus is a blood-feeding insect and a major vector of Chagas disease, a parasitosis endemic in Latin America and affecting millions of people worldwide. In the absence of effective drugs and vaccines, the control of the insect population represents a promising strategy to reduce the diffusion of the disease. Yet, genetic and functional studies in Rhodnius are extremely challenging due to its feeding habit and life cycle. To overcome these limitations, researchers have previously developed two stable cell lines derived from Rhodnius embryos. In this study, we provide the first characterization of the genes expressed in these cell lines. We found that, while the two cell lines share many expressed genes, each of them also has distinct gene expression patterns pointing to two different cell types with specialized functions. These differences likely affect the way they respond to stress and regulate biological processes. Our findings provide an important resource for researchers studying Rhodnius prolixus and other insect vectors, helping advance our understanding of the genetic and molecular mechanisms that control the insect development and mediate the interactions between insect vectors and the parasites they transmit

Nature-Based Solutions are increasingly promoted to address current urban challenges. While their potential effects on vector-borne disease risks have been documented, data on Aedes albopictus, a known arbovirus vector, remain limited in France. A previous study showed that urban vegetation moderately increases the abundance of adult mosquitoes of this species, but the monitoring period lasted only six months. Using ovitraps, we monitored Ae. albopictus egg density dynamics over multiple years (2022 to 2024) and analysed its environmental predictors in various urban environments. We included lagged meteorological variables, land cover metrics, and the cumulated egg densities recorded in the previous weeks as environmental predictors. Both parametric (GLMM) and non-parametric (Random Forest) models were fitted to weekly egg counts per trap. Our findings highlight that (i) egg density dynamics were related to how vegetation classes structured the landscape, (ii) growing degree days and cumulated number of eggs recorded in specific lagged time windows were the main contributors to egg density, and (iii) the non-parametric and parametric models performed similarly in terms of prediction accuracy.

Fasciolosis caused by Fasciola hepatica is an economically important disease in sheep and cattle. Knowledge of the population genetic structure of F. hepatica is important for understanding gene flow and informing disease control. In the present study, we designed, developed, and validated a multilocus sequence typing (MLST) scheme based on six markers. These markers were selected by aligning newly sequenced whole-genome sequence (WGS) data with available reference genomes and selecting variable regions with five or more single-nucleotide polymorphisms SNPs from different scaffolds of the F. hepatica reference genome Fasciola 10x pilon (GCA_900302435.1). Twenty markers were initially identified, of which 12 were multiplexed for deep amplicon sequencing after validation on worm and faecal eggs DNA; six markers were ultimately retained for downstream population genetics analysis. These markers were used to investigate population genetic structure in 15 cattle- and 27 sheep-derived F. hepatica populations in UK. A total of 53 unique alleles from six MLST markers were identified from 30 faecal (cattle = 13, sheep = 17) and 12 adult worm (cattle = 2, sheep = 10) populations. Shared alleles were observed in sheep- and cattle-derived populations. The highest allelic variation was observed in the Scottish Borders, Southern Scotland, and South-West England, and the lowest in North-West England. Minimal genetic differentiation was observed between cattle- and sheep-derived populations, with most genetic structuring within rather than between populations. Five markers showed high allelic polymorphism, whereas one marker showed low levels of allelic polymorphism, highlighting the importance of multilocus approaches. Overall, this six MLST-marker panel provides a tool for population genetic studies, revealing high gene flow and clonal expansion of F. hepatica across hosts and regions in the UK.

Soil-transmitted helminth (STH) infections are a major public health burden, and there are programmes of mass drug administration that attempt to ameliorate the harm that they cause. There has been increasing use of genomics to study STH infections and other parasitic nematodes, with particular interest in whole genome sequencing (WGS). For such studies, samples are commonly stored frozen, but in settings where these infections are endemic this can be difficult, and so there would be advantages to having ambient temperature storage methods. We investigated two ambient temperature storage methods - FTA cards and DESS buffer - for infective larvae of the rat parasites Nippostrongylus brasiliensis and Strongyloides ratti, prior to DNA extraction and then WGS. Our results showed that for individual larvae stored on FTA cards or in DESS buffer, this resulted in a lower proportion of sequence reads that mapped to the reference genomes, compared to the frozen control samples. Generally, for individual larvae, DESS-storage resulted in better sequencing results than FTA-storage. However, for pools of 10 or 50 larvae, then these ambient temperature storage methods generally resulted in comparable sequence read mapping to the frozen control samples.

Evidence-based decision making on malaria vector control strategies increasingly rely on triangulation of data which requires informatics systems that can integrate data from complex, multi-stage studies involving mosquitoes. This manuscript describes a performance evaluation of an extended version of the generic schema underpinning the VBDs360 platform, specifically improved to accommodate multiple distinct entomological assays spanning the field, insectary and laboratory. The utility of this extension, with respect to high-fidelity data linkage and robust sample traceability across complex entomological workflows, was evaluated through a case study conducted in southern Tanzania. Wild female mosquitoes were collected from 40 locations across a >4,000 km{superscript 2} area and then reared through multiple generations in an insectary before derived iso-female lineages were tested for phenotypic susceptibility to a pyrethroid insecticide. Such multi-generational lineages (F to F where n [&ge;] 2) were propagated to prevent non-heritable maternal effects on phenotype and produce enough progeny for standard WHO susceptibility assays. All samples were subsequently archived in a molecular laboratory, where all F specimens were tested for sibling species identity. A paper-based implementation of the extended schema enabled successful integration of 77,017 lines of data distributed across 6 different tables that spanned 3 distinct field, insectary, and laboratory workflows, implemented by three different teams working in different locations. At each step, fully independent and redundant primary and secondary keys enabled high fidelity error correction and sample tracing. Consistently perfect linkage between assay design and sample sorting data was achieved for F0 wild-caught adults, with 100% of 66,108 record successfully linked between field capture and morphological categorization. This complete traceability extended to the propagation of derived Fn lineages, with all 100 and 243 records from 9 adult-derived and 13 larval-derived lineages, respectively, correctly linked. Insecticide susceptibility phenotype further confirmed 100% linkage for 5,654 records between exposure history and recorded mortality outcome data in the insectary. Although such cross-cleaned linkages to sample analysis and storage data recorded by the laboratory team were not entirely perfect and could be improved, they were nevertheless of very high fidelity (97.3% (1967/2,022) for F0 samples and 99.3% (437/440) for Fn samples). Overall, this pilot application of the extended generic schema ensured robust data provenance and minimized transcription errors in this complex study distributed across multiple teams and locations. These findings demonstrate how this generic informatics framework may be scaled and adapted to support data integrity across diverse, large-scale, multi-team entomological research workflows.

The nuclear, internal transcribed spacer (ITS) and mitochondrial cox1 markers are widely used to differentiate Schistosoma haematobium from its livestock counterparts, S. bovis and S. curassoni. Schistosoma isolated from humans that have ITS and cox1 markers from livestock schistosomes are typically inferred as zoonotic infections, those with mixed species, heterozygous ITS are classified as F1s or recent hybrids, while those with discordant ITS and cox1 markers are considered to reflect older hybridization events. We evaluated the reliability of this classification scheme by genotyping ITS and cox1 from 132 parasites isolated from human urine, and from 37 adult schistosomes collected from cattle at 14 Nigerian locations. We also genome sequenced each sample to empirically determine livestock schistosome ancestry. ITS/cox1 genotyping suggested extensive recent hybridization and zoonotic infection. Among parasites from humans, 10.1% carried both S. curassoni and S. haematobium ITS, consistent with F1 or early generation hybrids, 21% had livestock schistosome markers at both cox1 and ITS suggesting zoonotic infection, while 13.7% carried S. bovis cox1 alongside mixed S. curassoni and S. haematobium ITS, suggesting more complex ancestry. Genome sequencing revealed a very different picture. All parasites from humans formed a tight cluster regardless of ITS or cox1 genotype, while all worms from cattle were well differentiated. We found no schistosomes containing 50% livestock parasite ancestry consistent with F1s. Instead, we observed regionally varying levels of S. bovis introgression, with modest levels in southern Nigeria (mean = 4.9%) and low levels in northern Nigeria (mean = 0.06%). These results demonstrate that: (i) two-locus genotyping is uninformative for detecting zoonotic infection or recent hybridization between S. haematobium and livestock schistosomes and (ii) previous data generated using this approach requires reinterpretation. These findings reveal the limitations of widely-used approaches for documenting zoonotic infection and hybridization between S. haematobium and livestock schistosome species.

Leishmaniasis, a climate-sensitive zoonotic neglected tropical disease, is transmitted by Phlebotomine sand flies and closely linked to socio-economic inequities. Understanding its spatio-temporal dynamics under environmental and social change is critical for effective control. A machine learning framework (XGBoost) was developed to map the global and European distribution of leishmaniasis, incorporating climatic indicators, land cover, elevation, and socio-economic indices (Human Development Index, AROPE). For Europe, five proven vector species (Phlebotomus perniciosus, P. ariasi, P. perfiliewi, P. neglectus, and P. tobbi) were modelled alongside cutaneous and visceral leishmaniasis. Across both analyses, land use features, particularly shrubland and forest cover, had the greatest explanatory power, reflecting their role in providing microclimates and vertebrate hosts for sand flies. Climatic factors, notably mean temperature of the coldest quarter and humidity of the warmest/driest quarters, were also influential, as these facilitate sand fly survival. Socio-economic predictors consistently improved model performance, confirming the role of poverty and inequity as determinants of disease distribution. Globally, leishmaniasis risk increased by ~17% since the 1990s, with Africa, Asia, and the Americas experiencing the greatest rise. In Europe, modest continental-scale increases (CL +1.28%; VL +2.47%) masked strong sub-national heterogeneity, including northward expansion of visceral leishmaniasis and increases in cutaneous leishmaniasis in southern and eastern regions. Sand fly projections indicated expansion of warm-adapted species (P. ariasi, P. perniciosus, P. neglectus) and contraction of species preferring cooler, more humid niches (P. perfiliewi, P. tobbi). These findings highlight climate change, land use, and inequity as interacting drivers of leishmaniasis, emphasising the need for enhanced surveillance, integrated vector management, and targeted support for vulnerable populations, including refugees and migrants.

BackgroundControl of Aedes aegypti, the primary vector of dengue and other Aedes-borne viruses, is challenged by insecticide resistance, limited efficacy of existing tools and the large and widespread epidemics. Targeted Indoor Residual Spraying (TIRS), a modification of traditional indoor residual spraying focused on Ae. aegypti resting sites, has demonstrated promising results, yet its indirect community-wide effects remain underexplored. Methodology/Principal FindingsWe conducted an entomological cluster-randomized controlled trial in Iquitos, Peru, to evaluate the direct and indirect entomological impacts of TIRS using pirimiphos-methyl. Thirty clusters were randomized to receive either TIRS (15 clusters, 898 structures) or standard Ministry of Health vector control activities (15 clusters, 1,018 structures). Aedes aegypti indoor densities were assessed in the 45 days pre-intervention and at four time points up to 255 days post-intervention using Prokopack aspiration. Generalized linear mixed models with a negative binomial link were used to estimate incidence rate ratios (IRRs) and calculate efficacy (1-IRR) for houses that received TIRS (direct effect) and untreated houses in TIRS clusters (indirect effect). Direct efficacy reached 96% at 15 days post-spraying and remained significant (40%) at 255 days post-spraying. Indirect efficacy reached 69% at 15 days and declined to 7% by 255 days post-spraying. Despite only 57% household-level TIRS coverage, both direct and indirect impacts on Ae. aegypti were significant during early post-intervention surveys, and after 8 months in TIRS clusters. Conclusions/SignificanceTIRS provided substantial and sustained reductions in indoor Ae. aegypti density, including measurable indirect effects in untreated homes within intervention clusters. These findings demonstrate the entomological value of TIRS even at moderate coverage levels and highlight its potential for both preventive and reactive vector control programs and should be considered for implementation by Ministries of Health in dengue-endemic urban settings as well as by the U.S. military when deployed to tropical or subtropical locations.

BackgroundMalaria remains a major public health challenge in Uganda, particularly in rural areas where access to conventional vector control tools is limited. Communities often use locally available plants as mosquito repellents, but documentation of the specific plants used, their utilization levels, and application methods in the Rwenzori region are limited. This study aimed to identify the types of plants used locally to repel mosquitoes, assess the level of utilization of plant-based mosquito repellents, and determine the methods of application employed by communities. MethodsA community-based cross-sectional study was conducted from June to December 2024 in the seven districts and one city of the Rwenzori region, Western Uganda. Multi-stage sampling was used to select 173 household heads. Data were collected using a pre-tested, translated (Runyoro, Rutooro, Lukonzo) KoboCollect questionnaire and analyzed descriptively with SPSS version 23. ResultsEighty-six percent of respondents reported using plant-based mosquito repellents, with 55% relying exclusively on plants. The most used plants were Cymbopogon citratus (citronella/lemon grass, 39.9%), Rosmarinus officinalis (rosemary, 25.7%), and Eucalyptus spp. (24.3%). The primary application method was planting repellent plants around the house (51.4%), followed by hanging injured plant parts in windows and doorways (28.4%). Other methods included burning or crushing plant parts and applying extracts/oils. ConclusionPlant-based mosquito repellents are widely used in the Rwenzori region. This study documents community knowledge and practices that could inform future integrated vector management strategies. Further research is needed to evaluate the entomological and epidemiological effectiveness of the plant repellents that are most used plants and the methods commonly applied.

Gastropods are a highly diverse and often overlooked taxonomic group of significant ecological and economic importance. Some terrestrial gastropods are critical pests of commercial agriculture and home gardens worldwide. Malacopathogenic nematodes offer an effective biological control method of managing pest slugs and snails as a natural enemy. Pellioditis (syn. Phasmarhabditis) hermaphrodita and Pellioditis (syn. Phasmarhabditis) californica are two species of biocontrol nematodes that have been commercialized, sold as Nemaslug(R) and Nemaslug(R) 2.0 respectively on three continents. Although there is interest in bringing Nemaslug(R) products to the US, they are currently not permitted due to limited knowledge on their North American distribution and effects on non-target and native species. In this study, we investigated the impact of P. hermaphrodita and P. californica on Ariolimax columbianus across two slug-host life stages, in laboratory infectivity assays. The objectives were to 1. determine whether P. hermaphrodita and P. californica nematodes impact survival of A. columbianus, and 2. evaluate whether there are differential effects on survival in juvenile and adult life stages of A. columbianus, in laboratory infectivity trials. We found that P. hermaphrodita caused significant mortality in A. columbianus with 100% mortality observed in both juvenile and adult slug hosts. The P. californica treatment had significant effects on the juvenile A. columbianus group only, with 80% mortality. By contrast, only 16% of unexposed control juveniles and 4% of control adult slugs died during the experiment. These results indicate that P. hermaphrodita and P. californica are lethal to the native, non-target Pacific banana slug (A. columbianus) under laboratory conditions, with mortality differing between juvenile and adult host life stages. Given the ecological importance of A. columbianus, these findings raise concerns for potential non-target effects of P. californica and P. hermaphrodita on terrestrial gastropod communities and emphasize the need for testing biocontrol agents against multiple life stages.

Mosquito-borne diseases such as dengue, chikungunya, and malaria remain major public health challenges in South Asia, particularly in Bangladesh, where mosquito repellents are widely used as primary preventive tools. This study presents a comparative evaluation of commonly used repellent products, including mosquito coils, liquid vaporizers and aerosols, DEET-based creams, and natural formulations, focusing on their efficacy, protection duration, and potential health risks. Efficacy was assessed using controlled laboratory methods, including chamber-based exposure and arm-in-cage tests against Aedes aegypti (Barnard & Xue, n.d.). Safety was evaluated through in vivo toxicological analysis in a rat model, incorporating clinical observations, hematological and biochemical profiling, and histopathological examination. The results indicated an overall mean effectiveness of 85%, with DEET-based creams demonstrating the highest efficacy (95%), while natural repellents showed the lowest (70%). Mosquito coils provided the longest protection duration ([~]10 hours) but were associated with the highest health risk due to combustion-related emissions. Vaporizers and aerosols offered moderate efficacy with moderate health risk. Toxicological findings revealed that coil exposure induced significant respiratory stress, elevated liver enzymes (ALT, AST), increased leukocyte count, and notable lung tissue damage. Vaporizer and aerosol exposure resulted in mild physiological changes, whereas DEET-based and natural repellents showed minimal systemic toxicity. Overall, the study highlights a clear trade-off between efficacy and safety across repellent types. These findings emphasize the need for informed product selection, stronger regulatory control, and the development of safer, cost-effective, and sustainable mosquito repellent technologies for effective vector control in endemic regions.

Tickborne diseases are a significant burden in many parts of the world. In the upper Midwestern United States, Lyme disease is the most common tickborne disease. It is carried by Ixodes scapularis. This vector can also transmit the pathogens causing anaplasmosis, babesiosis, ehrlichiosis, and several more tickborne diseases in this region. There is also concern for other tick species, such as Amblyomma americanum, that are expanding their ranges northward. We launched a citizen science passive tick surveillance program in 2024 to investigate tick species ranges in the upper Midwest, as well as the pathogens carried by I. scapularis. We received over 12,000 ticks in the first two years of this program, primarily from Wisconsin. While we received submissions of adult A. americanum outside of their endemic range, we did not see evidence of establishment in our study area. We measured pathogen prevalence in adult female I. scapularis (n=707) and observed 51% positivity for Borrelia burgdorferi, 9% for Babesia microti, 9% for Anaplasma phagocytophilum, and 3% for Ehrlichia muris eauclairensis. Multiple pathogens were identified in 14% of tested specimens, and significant associations were observed between B. burgdorferi and B. microti, and B. burgdorferi and E. muris eauclairensis. Pathogen prevalences varied across time and geography. Our results can begin to inform risk assessment for tickborne diseases in our region. A non-technical version of this document with interactive maps is available here: https://storymaps.arcgis.com/stories/8008c9d710b5400599f3c6cf88b2c546 Our online data dashboard is available here: redcap.link/TICS

Background Rabies is a zoonotic neglected public health problem associated with animal bites, especially domestic carnivores claiming 59,000 deaths annually predominantly in developing countries of Africa and Asia. There is a high risk of exposure among rural communities endemic with animal rabies where adoption of prevention strategies is minimal. This study determined the prevalence of suspected rabies exposure, associated factors, and delayed post-exposure care-seeking among animal-bite human cases in Busia district, Uganda. Methods: This was a cross-sectional study that involved 332 consecutively sampled animal bite human cases that occurred within the period 2023 to 2024. Data for the bite cases from records were collected using a data abstraction tool. In addition, interviewer-administered semi-structured questionnaires were used to collect data on sociodemographic, animal-related and environmental characteristics. Approximate bite locations were collected using Global Positioning System (GPS) coordinates via Kobo collect. Analysis was carried out in STATA 17 using mixed effects modified Poisson regression for factors associated with suspected rabies exposure. Results: The median age of the bite cases was 18 (IQR: 9-36) with the male gender predominantly affected. The prevalence of suspected rabies exposure was 53.6% (95% Confidence interval - CI: 46.8-60.3). Factors associated were urban versus (vs) rural residence (adjusted prevalence ratio-aPR: 1.04, 95%CI: 1.00-1.08), being bitten by a stray animal (aPR: 1.28, 95% CI: 1.22-1.35) and wild animal (aPR: 1.22, 95% CI: 1.14-1.30) vs domestic animal, vaccination status of the biting animal i.e. vaccinated vs unvaccinated (aPR: 0.76, 95% CI: 0.69-0.85), provoked vs unprovoked bites (aPR: 0.82, 95% CI: 0.79-0.86), and distance to nearest river ([&ge;]5km) vs <5km (aPR: 0.93, 95% CI: 0.87-0.99). The prevalence of delayed post-exposure seeking was 23.0% (95% CI: 16.5-31.1) among the suspected rabies exposures. Conclusion: The study reveals a high prevalence of suspected rabies exposure. Factors associated are multidimensional i.e. are of human, animal and environmental origin. The one health paradigm should be emphasized during routine surveillance of rabies-related cases. The study observed that 1 in 5 bite cases delayed to seek care post bite exposure. We recommend collaborations between sectors, routine vaccination and awareness campaigns, and monitoring of wild carnivore populations and environmental dynamics in rabies-related surveillance.

The ichneumonid subfamily Eucerotinae has been thought to be almost absent from the tropics, with the only known Afrotropical species found in Madagascar. We report the subfamily to be present in the mainland Afrotropics, and describe a new species, Euceros species 1 from Uganda and Cameroon (name not yet shown in preprint). The subfamily had likely not been observed in the mainland Afrotropics before due to low abundances and insufficient sampling. More Eucerotinae likely remain to be discovered in tropical Africa and Asia, although tropical America may genuinely have few eucerotine species. Much more extensive sampling will be needed before it is possible to make confident estimates of how eucerotine diversity is distributed globally.

Over the past decade, the global rise in invasive species has accelerated at an unprecedented rate, intensifying threats to ecosystems, human health, and economies worldwide. Newly invasive taxa, such as Tropilaelaps mites, are of particular concern for apiculture and agroecosystems. Despite growing concern about the spread of Tropilaelaps mites and other arthropods, limited resources are available to assess their invasive potential. We characterized 118 invasive arthropod species using available literature to identify key biological and ecological traits associated with invasive potential. We developed predictive generalized linear mixed models (GLMMs) to determine the traits most important for predicting invasive potential (number of invaded regions), and the top-performing models were subsequently applied to Tropilaelaps mercedesae. Several traits were identified as significant predictors of invasiveness, including the degree of human association, resilience at small population sizes, diet breadth, maximum annual number of generations, altitude range, and the interaction between human association and temperature range. Notably, T. mercedesae was predicted to be capable of invading 160 regions, ranking it within the top 10% most invasive species among those evaluated (12th out of 119), ranked just below the cosmopolitan Varroa destructor mite. These findings position T. mercedesae as a high-risk, yet under-recognized, invasive threat. Collectively, this demonstrates the power of predictive trait-based modeling to inform invasion risk prior to widespread establishment and underscores the urgency of reallocating resources toward surveillance, research, and proactive management strategies rather than relying on costly, often ineffective post-establishment eradication.

Progress towards elimination of onchocerciasis transmission is evaluated using entomological and serological indicators. The latter assesses anti-Ov16 IgG4 seroprevalence in children aged <10 years. A seroprevalence of <0.1% suggests that ivermectin Mass Drug Administration (MDA) can be stopped and post-treatment surveillance initiated, according to World Health Organization (WHO) guidelines. Surveillance of populations living in close proximity to vector breeding sites and first-line villages may mask spatial transmission heterogeneity. We mapped anti-Ov16 seroprevalence within a 25-km radius around a known Simulium damnosum s.l. breeding site in Asubende, Ghana, to assess its spatial patterns and their implications for serosurveillance. A cross-sectional survey was conducted in September-November 2024 in 30 settlements. Individuals aged [&ge;]3 years were recruited through convenience sampling. The Ov16 rapid diagnostic test (RDT) using dry blood spots was used to estimate overall and site-level seroprevalence. Generalized additive models were used to assess seroprevalence trends versus distance from the breeding site. Among 2,479 participants with valid RDT results, overall seroprevalence was 10.0% (95% CI: 8.9%, 11.3%) and increased with age. Seroprevalence varied across sites (0-24.4%) and declined with increasing distance from the breeding site. Among 584 children <10 years old, seroprevalence was 1.5% (95% CI: 0.7%, 2.9%). Adjusting for RDT sensitivity and specificity, seroprevalence in this age-group was 0.7%, (95% CI: 0%, 2.4%). Seropositive children were detected up to 18 km from the breeding site, but none were identified at it. The distance-related decrease in overall seroprevalence is consistent with spatial patterns in vector abundance at Asubende and implies higher cumulative exposure near the breeding site. The small number of children tested limited inference in this WHO target age-group. Detection of seropositive children away from, but not at, the breeding site highlights limitations of surveillance focused on first-line villages and supports broader spatial sampling to strengthen evidence for stop-MDA decisions. Author summaryAfter decades of onchocerciasis control using ivermectin, many countries hope to demonstrate that they have interrupted transmission of this vector-borne parasitic disease using serological surveys. It is unclear whether exposure to infection is spatially clustered around the riverine breeding sites of the blackfly vectors and therefore whether this is where serosurveillance should focus. To address this, we collected dried blood spots from 2,480 consenting participants aged 3-96 years old from 29 communities within a 25-km radius of a known blackfly breeding site in Asubende, Ghana. A rapid diagnostic test was used to test the blood spots for antibodies to the Onchocerca volvulus parasite. We found that overall seropositivity declined with increasing distance from the breeding site, which suggests that cumulative exposure is higher near the breeding site, where vector population is also high. However, seropositivity in children (3-10-year-olds, n= 584), which is indicative of recent transmission, was spatially distributed across the study area and found up to 18 km from the breeding site. These findings are relevant to serosurveillance sampling strategies intended to detect low levels of transmission, which could miss peripheral evidence of ongoing transmission if they are focussed at breeding sites and those villages closest to them.

RDL (Resistance to dieldrin) is a GABA-gated chloride channel that was first described as target of the insecticide dieldrin. Despite dieldrin being discontinued for decades because of its environmental per-sistence and health concerns, Rdl resistance mutations (A296S, A296G) continue at high frequencies in natural populations of the malaria mosquito Anopheles gambiae complex across Africa, suggesting a selective advantage. We have recently shown that RDL acts as a critical modulator of mosquito auditory sensitivity. Because acoustic recognition is essential for mate acquisition in An. gambiae, we hypothesized that these mutations confer a pleiotropic effect on mating success in the field, mediated through altered acoustic sensitivity, with potential consequences for sexual selection. We first provide laboratory evidence that resistance mutations enhance auditory behaviours of An. gambiae and show that the effect of environmental noise on mating success depends on the male Rdl genotype. We then conduct field collections in the city of Bangui (Central African Republic) and surrounding rural areas, revealing the presence of Rdl resistant alleles and their association with the urban environment, and within the city, with the noisiest locations. We also show decreased mating success of susceptible females with increasing noise levels, suggesting detrimental effects. Together, our findings support that Rdl resistance mutations enhance auditory function and mating success in acoustically challenging environments. We propose that this auditory advantage may contribute, together with other selective pressures such as cross-selection by other insecticides, to the persistence of these alleles in nature and may facilitate urban colonization by malaria vectors. Our study reveals, for the first time, an unintended evolutionary consequence of insecticide use, where a resistance mutation has been co-opted to enhance sensory performance and ecological adaptation, with significant implications for vector management strategies.

Close to 50% of all bird species are reservoirs of potentially pathogenic fungi, including those listed as priority by the World Health Organization. In Malawi, data on diversity, pathogenic potential, and ecological avian sources of medically important yeast are scarce. A cross-sectional study using a descriptive approach was conducted in Blantyre, Southern Malawi, to characterise medically important yeasts recovered from environments contaminated with excreta/guano from synanthropic pigeons. A total of 20 samples were collected from 4 peri-urban areas, which yielded 71 yeast isolates. To assess the pathogenic potential of the environmental isolates, we compared their phenotypic virulence traits with those of 21 clinical yeast isolates collected from referral hospital laboratories. Pichia kudriavzevii (39%) and Candida orthopsilosis (30%) were the commonly isolated species in the pigeon-guano-contaminated environments. Candida parapsilosis sensu stricto (29%) and Candida albicans (24%) constituted most of the clinical yeast isolates. Half of the species isolated in the pigeon-guano-contaminated environments were also identified among the clinical isolates. A majority of the environmental isolates showed virulence traits similar to or stronger than clinical isolates. The findings underscore the critical need for integrated surveillance under the One Health framework, especially in bird-inhabited spaces close to human settlements.