PLOS Neglected Tropical Diseases

BackgroundLymphatic filariasis remains a public health concern in many endemic regions, where chronic disease persists despite substantial reductions in transmission. In Ghana, more than two decades of mass drug administration have significantly reduced disease prevalence and transmission; however, chronic manifestations and gaps in community understanding continue to be reported in parts of the north. This study assessed infection status, chronic morbidity burden, and community knowledge in a rural setting in northern Ghana approaching elimination. Methodology/Principal FindingsA community-based cross-sectional study was conducted in Birifor, northern Ghana, from October 2024 to January 2025. A total of 261 residents aged ten years and above were selected using random sampling. Data collection included structured questionnaires, clinical examination for chronic disease, and night blood microscopy for the detection of infection. No microfilariae were detected (0/261; 0%). However, chronic lymphoedema was identified in five individuals (1.9%), all aged over 40 years. Awareness of the disease was high (95.8%), yet only 39.5% of participants demonstrated good community knowledge and perceptions and self-reported preventive practices. Misconceptions regarding transmission, particularly beliefs that the disease is hereditary or caused by spiritual factors, were common. Participation in mass drug administration was high (93.1%). Despite this, chronic disease imposed a notable socioeconomic burden: all affected individuals reported loss of income, and 60% reported additional household income loss due to caregiving. Conclusions/SignificanceThese findings suggest that transmission in the study area is likely very low, although residual infection cannot be excluded; however, chronic disease and gaps in community knowledge persist. Strengthening morbidity management, improving community education, and providing support for affected households are essential. Sustained surveillance and integrated approaches will be critical to prevent resurgence and support long-term elimination efforts. Author SummaryLymphatic filariasis, also known as elephantiasis, is a mosquito-borne disease that can cause long-term swelling of the legs, arms, or genitals. Global efforts have greatly reduced its occurrence, especially through repeated mass drug administration to afflicted communities. However, many people continue to live with chronic swelling caused by past infections, which can affect their ability to work and participate fully in daily life. In this study, we examined the current situation of lymphatic filariasis in a rural community in northern Ghana that has received many years of treatment. We tested people for active infection, assessed signs of chronic disease, and explored what community members know and believe about the disease. We found no evidence of active infection, suggesting that transmission is now very low. However, some individuals were still living with chronic swelling and reported loss of income, while households also experienced financial strain due to caregiving. Although most people had heard of the disease, many did not fully understand how it is transmitted. Our findings show that reducing transmission alone is not enough. Continued education, community support, and access to care are needed to address the long-term impact of the disease and support ongoing elimination efforts.

Background Nepal has set a goal to eliminate lymphatic filariasis (LF) by 2030. As of 2024, Nepal has stopped the mass drug administration (MDA) in 56 of the 64 endemic districts and completed two rounds of MDA in six districts with persistent LF ([&ge;]2% antigen prevalence) using the three-drug regimen of Ivermectin, Diethylcarbamazine, and Albendazole (IDA), exceeding 65% coverage. We subsequently conducted an Epidemiological Monitoring Survey (EMS) to assess the impact of the MDA in reduction of LF infection prevalence below the transmission threshold and examine the factors associated with it. Methods We conducted a cross-sectional EMS nine months after MDA in 12 evaluation units (EUs) across six districts, with two sites per EU. We recruited a total of 7,343 individuals aged [&ge;]20 years, sampled using multi-stage sampling, ensuring at least 300 blood samples collected per site. We collected data on demographics and MDA participation. We performed the LF antigen testing for all participants, followed by night blood microfilariae testing in antigen-positive individuals. Statistical analyses included non-parametric tests, Chi-square and Fishers Exact tests, and multivariable logistic regression to assess outcomes after adjusting for potential confounders. Results Nine of 12 evaluation units (EUs) recorded <1% microfilaremia, meeting the WHO threshold for passing EMS, while three EUs failed with [&ge;]1% prevalence in at least one site. Antigen and MF prevalence were 4.47% and 0.34%, respectively (ratio 13:1). Both Antigen and MF prevalences were significantly associated with female sex (AOR= 0.564, 95% CI: 0.441-0.721 and AOR = 0.326, 95% CI: 0.129-0.826 respectively) and participation in the most recent MDA round (AOR = 0.477; 95% CI: 0.385-0.591 and AOR = 0.089; 95% CI: 0.017-0.464 respectively). MDA uptake was influenced by age (<40 years, AOR = 0.72; 95% CI: 0.653-0.793), sex (female, AOR = 1.438; 95% CI: 1.29-1.603), cross-border residence (AOR = 0.616; 95% CI: 0.558-0.681), and occupation (agriculture and housewife, AOR = 1.144; 95% CI: 1.008-1.298). MF prevalence was also associated with younger age (<40 years, AOR = 0.211; 95% CI: 0.071-0.626). Conclusion The survey indicates progress toward LF elimination, with nine of twelve EUs achieving WHOs <1% microfilaremia threshold after two rounds of IDA-MDA. However, transmission persists in three sites, likely linked to poor MDA participation among specific subgroups--particularly males, younger adults, and cross-border populations. Strengthening MDA coverage and compliance across all demographic and occupational groups, with special focus on border areas, is essential to achieve LF elimination in Nepal.

BackgroundScrub typhus is a major yet neglected vector-borne disease in Thailand, where it has been nationally notifiable for over two decades. However, long-term changes in its epidemiology, including reporting rates, transmission intensity, disease severity, and seasonal patterns, have not been comprehensively characterised at the national level. MethodologyWe analysed 22 years of national surveillance data for scrub typhus in Thailand (2003-2024) using a latent process model that jointly fits reported cases with published nationwide seroprevalence data and antibody kinetics to estimate reporting rates and underlying transmission dynamics across all 77 provinces of Thailand. FindingsOver the 22-year study period, 143096 cases and 119 deaths were reported nationally. Estimated reporting proportion broadly mirrored transmission intensity, being higher in high-burden regions and lower elsewhere. A synchronous decline in detection was observed across all regions during the COVID-19 pandemic, followed by rapid rebound by 2024. After accounting for these reporting dynamics, the force of infection was highest in the northern provinces but also substantial in the northeast and south, with upward trends in some provinces. Susceptibility among older adults aged 65 and above increased progressively over the study period, reversing the pattern observed two decades earlier. Case-fatality in the 25-35-year reference group was low and declined from 0.14% (95% Credible Interval [CrI]: 0.06-0.29%) to 0.06% (95% CrI: 0.02-0.12%), but relative case-fatality remained consistently highest among adults above 65 across all periods. Three geographically distinct seasonal patterns were identified, all stable over time. ConclusionOver two decades, scrub typhus transmission in Thailand has been shown to extend well beyond its traditionally recognised northern focus, with substantial burden in previously underappreciated regions, while the demographic profile of those most affected has shifted progressively toward older adults. These findings support the need for regionally tailored surveillance, age-targeted clinical preparedness, and sustained investment in understanding the ecological drivers of transmission. Key messagesScrub typhus is a common but neglected cause of fever in Thailand, where it has been reported through the national surveillance system for over two decades. However, trends in reported cases can be misleading because they reflect not only true changes in transmission but also variation in diagnosis and reporting over time and across regions. We developed a model that combines surveillance data with seroprevalence surveys and antibody kinetics to separate true changes in transmission from variation in reporting, allowing us to estimate how transmission intensity, disease severity, and seasonal patterns have evolved from 2003 to 2024 across all 77 provinces. We found that substantial transmission occurs not only in the well-studied northern provinces but also in the northeast and south, where the disease has received less attention. Susceptibility has progressively shifted toward older adults, who also face the highest case-fatality, while three distinct seasonal patterns vary by region but have remained stable over time. These findings suggest that scrub typhus control in Thailand requires a shift from a predominantly northern focus toward regionally tailored strategies that account for local transmission timing, an ageing at-risk population, and the ecological drivers that sustain transmission in each setting.

Leishmaniases remain a significant global public health threat, with Leishmania amazonensis and Leishmania infantum representing the etiological agents of the cutaneous and visceral forms in the Americas, respectively. Building on our previous identification of Phospholipase A1 (PLA1) in Leishmania braziliensis, this study provides a comprehensive molecular, immunological, and biochemical characterization of PLA1 in L. amazonensis and L. infantum promastigotes. We analyzed PLA1 activity and expression, purified the recombinant enzyme from L. amazonensis, and validated protein expression using a specific anti-PLA1 serum. The major contribution of this research is the first description of the subcellular localization of a PLA1 within the Leishmania genus. Moreover, our results reveal an unprecedented association between PLA1 and lipid droplets within the parasites. This discovery is of particular interest as it provides the first evidence linking this enzyme to lipid storage organelles in Leishmania. Given that PLA1 is an established virulence factor in other trypanosomatids, these findings suggest a specialized role for the enzyme in parasite lipid metabolism and potentially in its pathogenic mechanisms, opening new perspectives for understanding Leishmania biology.

There is an ongoing Oropouche Fever (OF) outbreak in Brazil since 2024. There are dengue and chikungunya prediction models available, but none to help discriminate dengue, chikungunya, and OF. Objective: This study aims to develop and validate clinical prediction models for dengue, chikungunya, OF. Methods: This study uses surveillance data from Espirito Santo state / Brazil, from 2023-2025. Epidemiological investigations and biological samples were used to conclude cases as either (a) clinical-epidemiologically confirmed, (b) laboratory confirmed, or (c) discarded. The predictors were all data related to signs, symptoms, and comorbidities available in the notification forms. The analysis was performed using random forest regression models, one for each outcome, in development and validation datasets. Results: A total of 465,280 observations were analyzed, 261,691 dengue cases (56.6%), 18,676 chikungunya cases (4.0%), 12,174 OF cases (2.6%), and 179,115 discarded cases (38.6%). All three models had good discrimination and moderate to good calibration after scaling prediction. The models retained from 26 to 16 predictors each. Leukopenia and vomiting were the most discriminatory predictors for dengue, arthritis, arthralgia, and rash were the most discriminatory for chikungunya, and epidemiological features were the most relevant for OF. The dengue, chikungunya, and OF models had ROC AUC of 0.726, 0.851, and 0.896 in the validation set, respectively. Conclusion: This research identified predictors most discriminative between dengue, chikungunya, and OF. We developed and validated predictive models, one for each condition, with moderate to very good performance available at https://pedrobrasil.shinyapps.io/INDWELL/. One may use them in diagnostic work-up and arbovirus surveillance.

Social stigma surrounding chronic skin Ulcer leads patients to hide their wounds or delay seeking medical care. The aim of this study was to explore the types and causes of chronic skin ulcers among patients seen in the dermatology departments of two university hospitals in Burkina Faso. This was a cross-sectional, retrospective study covering an 11-year period, from 2013 to 2023. A review of consultation records allowed for the collection of sociodemographic and clinical data from 104 patients who were seen for chronic skin ulcers over the 11-year period, averaging 9 patients per year. The patients were primarily adults (n=60) and older adults (n=21). Leg ulcers were the condition observed in most patients (n=59). Eight cases of Buruli ulcer (7.69%) were identified among the 104 patients. Five of the eight cases, or 62.50%, were aged between 0 and 19 years. Half of the eight patients resided in Ouagadougou. These results highlight low utilization of dermatology services for chronic skin ulcers. Furthermore, indigenous cases of Buruli ulcer have been identified in Burkina Faso. Consequently, our findings call for the implementation of strategies focused on addressing social perceptions of these ulcers and on the screening and management of this disease.

Background/aims The World Health Organization (WHO) aims to eliminate schistosomiasis as a public health problem (EPHP) across 78 endemic countries by 2030. However, for low-prevalence settings that reach EPHP, guidance on managing transmission to maintain EPHP or move towards Interruption of Transmission (IoT) is limited, partly due to insufficient evidence on drivers of resurgence. In Uganda, some communities inland from Lake Victoria have achieved EPHP for Schistosoma mansoni but not progressed to IoT. This study explored whether routine, short-range travel to the highly endemic lake could sustain transmission in these settings. Methods We conducted a cross-sectional study in five Ugandan villages ~5 km from Lake Victoria. Parasitological data were collected using Kato-Katz and Point-of-Care Circulating Cathodic Antigen tests, alongside questionnaires on lake travel from 585 individuals aged 1-91 years. A structural causal model estimated the total and direct effects of travel frequency, activity type, water contact duration, and drug treatment history on infection. Bayesian regression models and counterfactual simulations predicted infection under hypothetical interventions. Results Reaching IoT in low-risk villages may be undermined by habitual, short-range travel to high-risk sites, driven by the nature and duration of lake contact. Daily lake travel caused a 1.7-fold increase in odds of infection, while occupational activities caused a 3.4-fold increase compared with no lake activity. Counterfactual analysis showed that removing lake contact duration most reduced infection risk among moderate-frequency travellers, while daily travellers showed smaller changes, and some transmission persisted among individuals with little or no lake contact. Simulations demonstrated that modifying lake contact behaviours could reduce individual infection risk but had limited population-level impact. Conclusion These findings indicate that targeting only high-risk villages or individual behaviours is unlikely to achieve sustained, wide-spread IoT, underscoring the need for integrated control strategies that account for mobility, behaviour, and local transmission ecology.

Background Venezuelan haemorrhagic fever (VHF), caused by Guanarito virus (GTOV), is a zoonotic disease endemic to the western plains of Venezuela. Despite decades of recognition, its epidemiology and clinical profile remain poorly characterised. Methodology We analysed individual level data from standardised case report forms submitted to the Venezuelan National Epidemiological Surveillance System between 2017 and 2024 for suspected VHF cases in Barinas, Apure, and Portuguesa. Demographic, clinical, and laboratory variables were examined to characterise temporal and geographical patterns and to define the clinical profile of VHF compared with endemic arboviral infections. Principal Findings Among 480 suspected cases, 72 (15.0%) were laboratory confirmed GTOV infections. Confirmed cases occurred predominantly in men engaged in agricultural or service related occupations, with the highest prevalence among individuals aged 46 to 90 years. A marked seasonal pattern was observed, with most cases occurring between September and January. The most frequently reported symptoms included headache, haemorrhage, sore throat, and diarrhoea. Compared with other endemic arboviral infections, GTOV was more strongly associated with headache, myalgia, sore throat, haemorrhage, and abdominal pain, delineating a distinct clinical phenotype relative to diseases caused by encephalitic alphaviruses, chikungunya virus, dengue virus, and Zika virus. The case fatality ratio among laboratory confirmed cases was 36.1% (95% CI: 25.1 to 48.3). GTOV infection was independently associated with mortality (adjusted relative risk [aRR] 3.66; 95% CI 2.28 to 5.87; p < 0.001), underscoring its substantial clinical severity. Conclusion GTOV remains endemically transmitted in western Venezuela, disproportionately affecting older men engaged in agricultural and service related occupations. Its seasonality and clinical phenotype, characterised by haemorrhage, sore throat, and gastrointestinal symptoms, highlight the need for clinical awareness and improved differential diagnosis, particularly in remote endemic settings with limited access to laboratory testing.

Background: Leptospirosis is a neglected tropical disease with substantial public health impact in Brazil, closely associated with socio-environmental vulnerabilities and climatic extremes. This study analyzed the epidemiological profile, spatiotemporal distribution, and climatic influences on leptospirosis incidence and lethality in Brazil from 2015 to 2024. Methods: An ecological time-series study was conducted using secondary data from the Notifiable Diseases Information System (SINAN). Variables included geographic region, probable infection environment, occupational, and educational level (ISCED-2011). The spatiotemporal correlation between disease incidence and El Nino-Southern Oscillation (ENSO) anomalies was assessed using the Oceanic Nino Index (ONI) and Spearman's rank correlation coefficient. Results: A total of 31,397 cases were notified, with an annual average of 3,140 cases. The South and North regions exhibited the highest incidence rates, while the Northeast and Southeast presented lethality rates above the national average (9.20%). A marked reduction in notifications occurred during the COVID-19 pandemic. Contaminations occurred predominantly in the domiciliary environment (64%). Rural workers (27.45%) and civil construction workers (18.63%) were the most affected occupational groups, with a higher incidence among illiterate and low-education populations. Climatic analysis revealed a positive spatial correlation between El Nino intensification and leptospirosis incidence in the South and Southeast, and a negative correlation in specific Northeastern states. Conclusion: The dynamics of leptospirosis in Brazil are complex and multifactorial, strongly influenced by macroclimatic variations and driven by deficits in basic sanitation and urbanization. Mitigating the disease burden requires sustained, region-specific public health strategies, targeted infrastructure improvements, and enhanced epidemiological surveillance to address underreporting.

Introduction: Following a large chikungunya outbreak during 2006 to 2008, Sri Lanka did not report any outbreaks for a 16 year period until end of 2008, possibly due to population immunity. Therefore, understanding baseline immunity prior to outbreaks is crucial to inform implementation of vaccine strategies. Methods: We assessed the age stratified seroprevalence for chikungunya in an urban (n=816) and a semi urban (n=380) community in Colombo, Sri Lanka, from September to November 2024, prior to the commencement of the large chikungunya outbreak, in December 2024. Sociodemographic, socioeconomic and clinical data were collected and chikungunya specific IgG measured in serum samples. Results: Of 1196 participants, 410 (34.3%) were chikungunya IgG seropositive. Seroprevalence was significantly higher in urban populations compared with semi urban populations (39.6% vs 22.9%; p<0.001) and increased significantly with age in urban areas but not in semi-urban areas. Living in an urban area was the strongest independent risk factor of chikungunya seropositivity (aOR 7.48, 95% CI 4.05 to 13.81; p<0.001), consistent with the higher population density, poor housing conditions and overcrowding observed in that setting. The use of mosquito nets was independently associated with reduced risk of seropositivity (aOR 0.50, 95% CI 0.27 to 0.93; p=0.029). Almost no individuals aged <16 years had evidence of prior infection (0.55%), indicating minimal transmission in the preceding 16 years. In the urban cohort, seropositivity was significantly associated with diabetes, central obesity, overweight, and hypertension. Conclusions: There appears to have been minimal chikungunya transmission in the 16 years preceding the 2024 outbreak, with a large population susceptible to chikungunya. Higher seroprevalence in urban populations highlights the role of population density, overcrowding, and housing conditions as key drivers of transmission.

Dengue transmission is strongly influenced by climatic conditions that affect mosquito population dynamics and virus circulation. In Southern Brazil, where dengue historically occurred at low levels, recent climatic anomalies may be contributing to the expansion of Aedes vectors and an increase in local dengue incidence. This study investigated the spatiotemporal association between climatic variables, Aedes mosquito infestation and dengue cases in Porto Alegre (Southern Brazil, 2018 to 2025). Entomological, surveillance and climatic data were analyzed using Morans I and LISA for spatial association, Kendall correlation, polynomial regression and LASSO to identify relevant drivers and develop predictive models of mosquito infestation and dengue incidence. A strong spatial association between Aedes aegypti and Aedes albopictus was observed, with persistent local clusters detected across all years. Annual climatic variables were associated with mosquito abundance in several districts. Overall, rainfall frequency had a stronger effect on Aedes aegypti abundance than accumulated rainfall. Temperature and lagged infestation indices showed strong association with both species and dengue incidence, with effects observed up to four weeks prior. Predictive models demonstrated good agreement between observed and predicted values, particularly within low to moderate infestation levels. Lagged variables were consistently retained in both mosquito infestation abundance and dengue incidence models, highlighting the importance of temporal predictors for anticipating vector dynamics and dengue risk. This approach is generally applicable for predicting Aedes infestation and disease incidence and emphasizes the importance of integrating entomological and climatic surveillance data to improve anticipation and detection of dengue risk periods and support more effective public health interventions.

BackgroundSchistosomiasis remains a major public health challenge in sub-Saharan Africa. Recent World Health Organization (WHO) guidance calls for community-wide treatment and fine-scale data to optimise preventive chemotherapy (PC) strategies, yet the practical implications for resource allocation by health ministries are unclear. MethodsWe analysed epidemiological and cost data from Ethiopia and Zimbabwe to compare survey designs and five implementation scenarios. Scenarios varied by data source, administrative unit of implementation, WHO guidance on PC strategies. Outcomes were target population, praziquantel needs, and delivery costs. ResultsGeostatistical surveys reduced sample size by up to 90% and survey costs by [&ge;]72% compared with a design-based approach, while increasing spatial coverage. Applying updated WHO guidance expanded eligibility to pre-school-aged children and adults, and in one scenario increased treatment needs by 72% in Ethiopia and 262% in Zimbabwe. Correspondingly, praziquantel requirements and delivery costs were driven primarily by expanded age eligibility rather than geographic coverage. ConclusionsGeostatistical surveys provide substantial efficiency gains for impact assessments, enabling cost-efficient, granular targeting. However, implementing 2022 WHO guidance was the dominant driver of increases in programme scope and resource needs, underscoring the importance of accurate fine-scale data to guide efficient planning and budgeting toward elimination goals. Author summarySchistosomiasis control programmes are required to use finer-scale data and updated World Health Organization (WHO) guidance to decide where and how often to deliver praziquantel. We analysed national schistosomiasis data and programme costs from Ethiopia and Zimbabwe to compare different approaches to impact assessment surveys and to estimate how treatment needs change under alternative decision rules.We found that model-based geostatistical surveys can reduce the number of people that need to be sampled and the cost of surveys while providing more detailed information for planning at sub-district level. However, when we applied the 2022 WHO schistosomiasis guidance, expanded eligibility (including adults and pre-school-age children and a lower threshold for community-wide treatment) substantially increased the number of people needing treatment. In our scenarios, expanded eligibility drove much larger increases in praziquantel requirements and delivery costs than changes in geographic coverage. Our findings help health ministries to anticipate the operational and budget implications of updated guidance and highlight why accurate fine-scale data are essential for equitable and realistic planning toward elimination.

Background Urogenital schistosomiasis (UGS), caused by Schistosoma haematobium (S. haematobium), disproportionately affects women in sub-Saharan Africa and can lead to haematuria, anaemia, and urinary tract morbidity. Data on the prevalence in women of reproductive age remains limited in contrast to infection among school-aged children in Kenya. This study assessed the prevalence of UGS and its socioeconomic determinants among women in Kilifi County, Kenya. Methods: Urine samples (20-50 mL) were collected from each participant over three consecutive days. Day-one samples were tested for haematuria, proteinuria, nitrites, leukocytes, and pregnancy using dipsticks. On the other hand, 10 mL of urine was examined for S. haematobium eggs via urine filtration on all three days. Results: A total of 599 women aged 15-50 years were enrolled, with complete data available for 336. The mean age was 33 years; 57.7% were <35 years. Most participants were from rural Magarini Sub-county (63%) and engaged in crop farming (62.5%). Primary education was the highest level attained by 59.8% of participants. Frequent contact with stagnant water was reported by 92%. The overall prevalence of S. haematobium infection was 13.7% (95% CI: 10.2-17.8), higher in Magarini (14.9%) than in Rabai (12.0%), though not statistically significant. Younger age, primary education, and frequent water contact were associated with higher infection rates; however, after adjustment for covariates, haematuria showed the strongest independent association with infection. Women with haematuria were 25.2 times more likely to be infected (OR: 25.24, 95% CI: 7.07-82.63, p < 0.001); multivariate analysis confirmed haematuria as the sole significant predictor (OR: 20.83, 95% CI: 5.45-79.57, p < 0.001). Conclusion: UGS prevalence among women in Kilifi County is substantial, with variation between sub-counties. Haematuria demonstrated the strongest independent association with infection and may serve as a simple, non-invasive diagnostic marker. These findings underscore the pressing need for the integration of UGS screening into the reproductive health services and targeted interventions. Authors Summary UGS, caused by the parasitic worm Schistosoma haematobium, is a neglected tropical disease and remains a major public health problem in sub-Saharan Africa. Although control programmes in Kenya primarily target school-aged children, women of reproductive age are frequently exposed through daily water contact and may develop chronic urinary and reproductive health complications. However, data on the infection burden among adult women are limited. In this study, we assessed the prevalence of urogenital schistosomiasis and associated risk factors among women aged 15-50 years in Kilifi County, Kenya. Urine samples were collected over three consecutive days and examined for parasite eggs and indicators of urinary tract disease. We found that urogenital schistosomiasis affected more than one in ten women in the rural sub-counties where the study was conducted. Haematuria was strongly associated with infection and remained the most reliable predictor after accounting for other social and behavioural factors. These findings demonstrate that UGS is an under-recognised health issue among women and highlight the potential value of simple urine-based screening tools. Integrating UGS screening into existing reproductive health services could improve early detection and contribute to more inclusive disease control strategies.

Helminthiases remain a major global health burden, and limitations of current anthelmintic therapies highlight the need for new pharmacological targets. In this study, we examined the effects of ion-channel and cytoskeletal modulators on bovine lung protoscoleces (PSCs) of Echinococcus granulosus sensu lato. Compounds acting on ion channels (praziquantel, amiloride, and amlodipine) and cytoskeletal components (albendazole and cytochalasin D) were evaluated using a semi-automated motility assay, methylene blue exclusion to assess viability, and scanning electron microscopy (SEM) to characterize structural damage. All compounds produced concentration-dependent reductions in PSCs motility. Amlodipine was the most potent inhibitor of motility, whereas praziquantel and cytochalasin D produced pronounced tegumental alterations and strong correlations between motility impairment and parasite death. In contrast, amiloride markedly reduced motility with comparatively minor effects on viability, indicating a primarily paralytic effect. Cytoskeletal disruption induced severe structural damage and parallel declines in motility and viability. SEM analysis revealed extensive tegumental collapse, loss of glycocalyx, and microtrichial damage in PSCs exposed to cytoskeletal and calcium-modulating agents. These findings highlight cytoskeletal organization and calcium-dependent ion fluxes as key physiological vulnerabilities in E. granulosus. Comparative analysis of these pharmacological targets provides mechanistic insight into how disruptions in cytoskeletal dynamics and cation homeostasis compromise parasite motility and survival.

Ticks impose major health and economic losses on the livestock sector of Pakistan, yet uncertainty-aware maps of tick burden remain scarce. We focused on the two most common disease transmitting tick species, Rhipicephalus microplus and Hyalomma anatolicum, to produce exposure-adjusted district-level abundance estimates and predictions for unsampled areas in Punjab and Khyber Pakhtunkhwa (KPK). We compiled heterogeneous tick count records and standardized them per 100,000 animals. District-level climate and physiographic covariates were summarized via principal components analysis. Bayesian spatial models were fit in R-INLA using Gaussian likelihoods and BYM2 over a hybrid adjacency matrix. Competing non-spatial and spatial models were compared, and the best model was used to generate posterior predictions and 95% credible intervals for unsampled districts. Spatial models outperformed non-spatial alternatives and calibrated well. Model robustness was confirmed through eight independent 80/20 train-test splits, showing strong generalization with consistent predictions across seeds. For unsampled areas, R. microplus exhibited a pronounced north-south gradient with high predicted means but wide intervals in the northern highlands, indicating information gaps. H. anatolicum predictions were highest and most precise in southern Punjab. Sensitivity analysis highlighted a dominant spatial component, with modest contributions from PC1 and PC2. The Bayesian spatial models using the Besag-York-Mollie framework delivered comparable, exposure-adjusted tick abundance maps while quantifying uncertainty to guide surveillance. Results suggest a need for immediate control in confirmed hotspots and recommend targeted field sampling in high-uncertainty districts. The workflow generalizes to other vectors, pathogens, and regions for evidence-based livestock health planning.

Background: Spotted fever is caused by spotted fever group rickettsiae (SFGR) belonging to the genus Rickettsia. Transmission to humans is primarily via the bite of infected ticks. Being a vector-borne disease, the occurrence of spotted fever is related to factors that allow the vector to thrive. This spatio-temporal analysis gives an insight into the distribution of cases and correlation with seasonality. Methodology: A suspected AFI patient was considered spotted fever positive if either serology (ELISA/IFA) or molecular assay (Nested PCR/qPCR) was tested positive. Demographic data of confirmed cases were included for the analysis. Results: In the 18-year dataset, a total of 2153 suspected patients were tested for spotted fever, of which 516 (24%) were positive. On spatio-temporal analysis, Vellore district reported 39.9% of cases, Chittoor 38.8%, Tirupattur 12.5%, Ranipet 4.5%, and Tiruvannamalai 4.3%. Maximum spotted fever cases were reported between the months of September to March, with a peak in January. Children below 10 years and housewives were at risk of spotted fever. Conclusion: The findings of this retrospective analysis highlight the importance of considering spotted fever group rickettsioses in patients presenting with acute undifferentiated febrile illness, particularly children aged <10 years, from areas with higher spatial clustering, during or following the monsoon season.

The wMel strain of the insect endosymbiont Wolbachia reduces the potential for Aedes aegypti to transmit mosquito-borne viruses such as dengue (DENV). Field trials that have introgressed wMel into Ae. aegypti populations have shown this approach significantly reduces dengue incidence. In a laboratory setting some wMel-Ae. aegypti develop infectious saliva following a viremic blood meal. Additionally, studies have demonstrated that exposing wMel-Ae. aegypti to heat treatment, particularly during the larval stage, reduces wMel density in key tissues such as the ovaries, midgut and salivary glands. Here we build on these studies, using viremic blood collected from 13 dengue inpatients at the Hospital for Tropical Diseases in Ho Chi Minh City (Viet Nam), to assess how temperature affects the protection afforded to Ae. aegypti by wMel. We found that, compared to wMel-Ae. aegypti reared at 28 {+/-} 4{degrees}C, those reared at 31 {+/-} 4{degrees}C developed infectious saliva more frequently, but the risk of this occurring was still reduced compared to WT mosquitoes reared at the same temperature. Heat treatment reduced the density of wMel in all tissues tested, decreased the magnitude of wMels protection against DENV replication in the head/thorax, and significantly increased the amount of DENV replication in wMel-Ae. aegypti. When comparing cohorts of wMel-Ae. aegypti that did or did not develop infectious saliva, DENV levels in the head/thorax were associated with increased odds of mosquitoes developing infectious saliva, but wMel density was not. Overall, these findings show that elevated rearing temperatures increase the risk of patient-derived DENV breakthrough infections in wMel-Ae. aegypti, potentially due to increased DENV replication in these mosquitoes. This limitation suggests it would be prudent to increase surveillance in regions using wMel for dengue control when daily mean temperatures remain above 30{degrees}C for multiday periods. Author SummaryThe mosquito species Ae. aegypti can be infected with the bacterium Wolbachia (wMel strain), reducing its capacity to transmit viruses like dengue (DENV). Wolbachia is now being used as a biocontrol tool to reduce the burden of dengue in communities. However, some mosquitoes with Wolbachia can still transmit DENV. Here we utilised a natural infection model using dengue patient-derived blood to examine how temperature may increase the risk of DENV transmission occurrence in mosquitoes with wMel. Mosquitoes with wMel were more likely to transmit virus when reared at an average temperature of 31{degrees}C compared to those reared at an average temperature of 28{degrees}C but these mosquitoes still had a lower risk of developing infectious saliva compared to their wMel-free counterparts. Higher temperatures reduced the amount of wMel in mosquito tissues and increased the amount of DENV replicating in the head/thorax. Increasing levels of DENV RNA in these tissues were found to be associated with increased risk of mosquitoes with wMel developing infectious saliva. This finding indicates surveillance is warranted in high temperature settings or during heat waves, to monitor for changes in wMel frequency and DENV infection in Ae. aegypti.

Dengue virus (DENV) infection is a major global health threat, affecting more than half of the worlds population. Severe dengue is a life-threatening condition characterised by systemic bleeding, vascular leakage, and interstitial fluid accumulation that can progress to hypovolaemic shock. Circulating DENV non-structural protein 1 (NS1) has long been implicated in driving vascular hyperpermeability through its disruptive effects on endothelial cell junctions and the glycocalyx. The lymphatic system, which runs alongside the vascular network, plays a critical role in resorbing and recirculating interstitial fluid and immune cells extravasated from blood vessels. Despite its importance in maintaining tissue fluid homeostasis, the impact of dengue disease on lymphatic vessels has not previously been explored. Here, we present the first evidence that DENV-2 NS1 induces marked hyperpermeability in lymphatic endothelial cells, as measured by transendothelial electrical resistance, and impairs lymphangiogenesis in vitro. These effects were not attributable to changes in cell viability, morphology, or metabolic activity, as assessed by live/dead and metabolic assays and image analysis. Instead, we observed a defect in lymphatic endothelial cell migration, measured by scratch assay, which may underlie the reduced lymphangiogenic potential. Bulk RNA-seq, immunocytochemistry, and advanced image analysis further demonstrated pronounced reorganisation of cell-cell junctions, the cytoskeleton, and focal adhesions. Notably, junctional proteins including VE-cadherin, ZO-1, and Claudin-5 were not downregulated but instead displayed disorganised distribution along the cell junctions or aberrant cytoplasmic localisation. These structural disruptions became even more pronounced under flow conditions produced using a microfluidic system. Together, these findings demonstrate for the first time that DENV-2 NS1 directly disrupts lymphatic endothelial cell function, leading to junctional disorganisation and hyperpermeability. Such impairment of lymphatic drainage may contribute to the pathophysiology of severe dengue. Author SummaryDengue is a rapidly expanding mosquito-borne disease that now affects many tropical and subtropical regions worldwide. Severe cases can lead to extensive fluid leakage from blood vessels, which causes tissue swelling and, in the most dangerous situations, shock. Although much research has focused on how dengue damages the blood vascular system, almost nothing is known about its impact on the lymphatic system, which is responsible for removing fluid from tissues and returning it to the bloodstream. Because both systems work together to maintain fluid balance, understanding how dengue affects lymphatic vessels is important for explaining why fluid accumulation becomes so severe in critical disease. In our study, we examined whether the viral protein NS1, which circulates during infection, directly affects the cells that line lymphatic vessels. We found that NS1 increases the permeability of these cells and reduces their ability to form new vessel structures. These effects were not caused by cell death but by disruptions in how the cells organise their junctions, internal scaffolding, and interactions with neighbouring cells. By showing that NS1 can directly impair lymphatic vessel function, our work identifies a previously overlooked mechanism that may contribute to fluid build-up in severe dengue and suggests new avenues for future therapeutic research.

Background Cutaneous leishmaniasis (CL) is highly prevalent in Ethiopia, including the Tigray region. However, there is a dearth of information on the levels of knowledge, attitude, and health seeking behavior among the communities in CL-endemic areas of Tigray region, northern Ethiopia. Objective This study aimed to investigate CL-related knowledge, attitude, treatment-seeking and prevention practices in disease-endemic areas of Tigray. Methods Between November and December 2022, a cross-sectional survey was conducted among communities living in seven districts of Tigray. A mixed sampling method was implemented. Data were collected using a pre-tested structured questionnaire and analyzed using SPSS 25 (IBM, Chicago). Results A total of 512 participants were included. Overall, 43%, 36% and 34% of participants had a good level of knowledge, a favorable attitude and a good treatment-seeking and prevention practices towards CL, respectively. However, nearly all participants did not know about CL transmission, about 25% perceived CL to be genetically acquired and about 67% believed it to be stigmatizing. Traditional medication was the preferred option over modern treatment for 63.3%. Rural dwelling participants (AOR = 1.60; 95% CI: 1.00-2.57) and participants living in households with CL episode (AOR = 10.19; 95% CI: 6.36-16.30) had good knowledge towards the disease. However, urban/ semi-urban residents (AOR = 2.17; 95% CI: 1.42-3.31) had favorable attitude towards CL. Gender (AOR = 1.49; 95% CI: 1.01-2.22) and education level (AOR = 0.39; 95% CI: 0.24-0.62) were significantly associated with treatment-seeking and prevention practices. Participants living in households with CL episode (AOR = 2.99; 95% CI: 1.96-4.57) had good treatment-seeking and prevention practices. Conclusion In this study, over one half of participants had poor knowledge about CL, nearly two-third of them had unfavorable attitude towards the disease and two-third of them had poor treatment-seeking and prevention practices. Residence and previous CL episode in households were determinants of respondents knowledge about CL and their attitude towards the disease. Level of education and living in households with CL episode were determinants of participants treatment-seeking and prevention practices. These findings support for an integrated intervention through health education focusing on CL transmission and preventive measures.

This study evaluated the effects of different Arachis hypogaea dietary preparations on parasite load, haematological indices, and physiological responses in Plasmodium berghei-infected mice. Forty-five albino mice were randomly assigned to five groups: normal control, infected untreated control, roasted groundnut, boiled groundnut, and a combination of roasted and boiled groundnut diets. Data were analyzed using one-way ANOVA at p > 0.05. Infection resulted in a high parasite load in the untreated group, with no significant difference compared to the boiled and combined diet groups. However, the roasted groundnut group showed a reduction in parasite load and relatively higher chemosuppressive activity, although differences were not statistically significant. White blood cell counts increased significantly following infection, and dietary treatments did not restore normal levels. Similarly, red blood cell counts and packed cell volume were significantly reduced in infected mice. The roasted groundnut diet moderately improved PCV compared to other treatments but did not restore it to normal levels. Weight loss was most pronounced in untreated mice, while roasted groundnut intake showed slight mitigation. No significant effects on temperature regulation were observed. Overall, A. hypogaea diets did not significantly improve parasitemia or haematological parameters, indicating limited therapeutic value in malaria management. ImportanceThis study is of significant importance due to its contribution to the ongoing search for accessible, affordable, and nutritionally based supportive interventions in malaria management. Malaria remains a major public health burden, particularly in sub-Saharan Africa, where increasing resistance of Plasmodium species to conventional antimalarial drugs continues to undermine control efforts. By investigating the effects of Arachis hypogaea (groundnut), a widely consumed and locally available food resource, this research explores a practical dietary approach that could complement existing malaria treatment strategies. In summary, this research is important because it bridges nutrition, parasitology, and public health, offering practical insights that could inform both scientific advancement and real-world malaria management strategies.