PLOS https://journals.plos.org/plosntds/ customercare@plos.org https://journals.plos.org/plosntds/feed/atom All PLOS articles are Open Access. https://journals.plos.org/plosntds/resource/img/favicon.ico https://journals.plos.org/plosntds/resource/img/favicon.ico 2026-05-16T02:37:20Z Max Hoekstra Kirby R. Lattwein Wendy W. J. van de Sande 10.1371/journal.pntd.0014291 2026-05-15T14:00:00Z 2026-05-15T14:00:00Z <p>by Max Hoekstra, Kirby R. Lattwein, Wendy W. J. van de Sande</p> The Neglected Tropical Disease mycetoma is a chronic, progressive infectious disease characterized by large tumor-like masses in subcutaneous tissues. It is endemic in rural areas of low- and middle-income countries, where it can lead to extensive health and socioeconomic burdens for the patient and their family. It is especially prevalent among male farmers. The disease can be caused by both bacteria and fungi, referred to as actinomycetoma and eumycetoma, respectively. Inoculation occurs when a skin breach allows the pathogen to enter the subcutaneous tissue. The pathogen can form grains, which make the disease more resistant to host defense and antimicrobial treatments. This review summarizes the current knowledge on the historical background and epidemiology of mycetoma, the characteristics of its causative agents, underlying pathophysiology, diagnostic approaches, available treatment strategies, preventive measures, and future prospects for disease management and research. Although the recognition of mycetoma as both a neglected tropical disease and a fungal priority pathogen has increased awareness and research interest, large knowledge gaps remain. Lester Gutiérrez Nadja A. Vielot Yaoska Reyes Roberto Herrera Christian Toval-Ruiz Javier Mora Michael B. Arndt Rebecca Barney Robert K.M. Choy Filemón Bucardo Samuel Vilchez Sylvia Becker-Dreps Luther A. Bartelt 10.1371/journal.pntd.0013734 2026-05-15T14:00:00Z 2026-05-15T14:00:00Z <p>by Lester Gutiérrez, Nadja A. Vielot, Yaoska Reyes, Roberto Herrera, Christian Toval-Ruiz, Javier Mora, Michael B. Arndt, Rebecca Barney, Robert K.M. Choy, Filemón Bucardo, Samuel Vilchez, Sylvia Becker-Dreps, Luther A. Bartelt</p> Background <p><i>Giardia lamblia</i> (<i>Giardia</i>) is one of the most common intestinal parasitic infections globally, with an estimated 280 million symptomatic infections annually. In children from low- and middle-income countries (LMICs), <i>Giardia</i> is highly prevalent and has been associated with loss of intestinal barrier function, nutrient-metabolic dysregulation, and linear growth impairment, but specific mechanisms linking <i>Giardia</i> to these outcomes remain poorly understood.</p> Methods and results <p>We used data and samples from a subset of 76 children in a longitudinal birth cohort in Nicaragua to evaluate the natural history and geospatial distribution of <i>Giardia</i> infections, child growth outcomes (weight-for-age [WAZ] and length-for-age [LAZ] z scores), and relationships with established biomarkers of inflammation, intestinal damage, and growth-signaling. During the first 36 months of life, we tested 2,305 stools (1,903 surveillance stools and 402 diarrheal stools) for <i>Giardia</i> by qPCR. The incidence of <i>Giardia</i>-positive stools was 59.6 per 100 child-years. Any detection of <i>Giardia</i> was associated with a reduction in LAZ at 36 months of life (β:-0.16, <i>P</i> = 0.042). This effect increased when considering persistent or recurrent <i>Giardia</i> detections (β:-0.26, <i>P</i>=<0.001) as well as living in a high-density <i>Giardia</i> detection area (β:-0.44, P=<0.001). Among intestinal markers, <i>Giardia</i> was associated with lower median fecal neopterin (a marker of chronic intestinal T cell activation) at 24 and 36 months of age. Among serum systemic biomarkers measured at 24 months, <i>Giardia</i> detections were associated with indicators of intestinal epithelial cell damage (higher median Intestinal Fatty Acid Binding Protein (<i>P</i> = 0.002) and Anti-FliC IgA (P = 0.033), and reduced growth-signaling hormone (lower median Insulin-like Growth Factor (IGF-1) (<i>P</i> = 0.005).</p> Conclusion <p><i>Giardia</i> detection was negatively associated with linear growth in an exposure-dependent manner. Simultaneously, <i>Giardia</i> was associated with diminished serum growth-signaling hormones. Patterns of serum and fecal intestinal biomarkers suggest that <i>Giardia-</i>mediated epithelial disruption is dissociated from markers of intestinal inflammation.</p> Choo Hock Tan Praneetha Palasuberniam Lee Louisa Pernee Kae Yi Tan 10.1371/journal.pntd.0013859 2026-05-14T14:00:00Z 2026-05-14T14:00:00Z <p>by Choo Hock Tan, Praneetha Palasuberniam, Lee Louisa Pernee, Kae Yi Tan</p> Short-chain α-neurotoxins (SNTXs) are the principal neurotoxic components in several Asiatic cobras, including <i>Naja atra</i>, <i>Naja philippinensis</i>, and <i>N. samarensis</i>. Although structurally conserved, SNTXs exhibit marked antigenic variation that can limit the effectiveness of regional antivenoms used for snakebite envenoming in Asia. Here, we evaluated the immunoreactivity of the Philippine Cobra Antivenom (PCAV) and three regional products—<i>Naja kaouthia</i> Monovalent Antivenom (NkMAV), Neuro Bivalent Antivenom (NBAV), and Indonesian SABU—against a panel of cobra venoms and purified α-neurotoxins. PCAV bound strongly to homologous <i>N. philippinensis</i> SNTX but showed significantly lower cross-reactivity with SNTXs from <i>N. kaouthia</i>, <i>N. sputatrix</i>, and <i>N. atra</i>, and minimal recognition of marine elapid SNTXs and long-chain α-neurotoxins (LNTXs) of Monocled Cobra as well as King Cobra. Conversely, NkMAV and NBAV recognized mainland Asian SNTXs more broadly but reacted poorly with the Philippine cobra toxin. These differences were statistically significant in multiple comparisons <i>versus</i> the homologous venom control. Hierarchical clustering of normalized immunoreactivity delineated two major SNTX antigenic subtypes corresponding to Philippine versus continental Asian lineages. Peptide sequence analysis unmasked two distinct loop-II motifs (²⁸WWS–TII³⁷ and ²⁸RWR–YRT³⁷) associated with these divergent immunotypes. Phylogenetic reconstruction suggests that the Philippine cobras retain an ancestral SNTX motif, while Sundaic and East Asian cobras have diversified to employ another major SNTX form. Epitope prediction further revealed differences in surface exposure and accessibility, which helps explain the limited cross-neutralization among antivenoms. antivenoms. These findings demonstrate a clear antigenic dichotomy among Asian cobra SNTXs, which underlies species-specific antivenom effectiveness and highlights the need to incorporate representative SNTX variants into immunogen formulations to improve regional antivenom coverage. Paulo C. Manrique-Valverde Chloe M. Hasund Katrina A. Kelley Jorge-Eduardo Amaya-Romero Myriam Arévalo-Herrera Raphael Brosula Jose E. Calzada Stella M. Chenet Vladimir Corredor Angela M. Early Gustavo Fontecha David A. Forero-Peña Sócrates Herrera Justin T. Lana Margaret Laws Reza Niles-Robin Nicanor Obaldia Ana M. Santamaria Philipp Schwabl Sarah Auburn Daniel E. Neafsey 10.1371/journal.pntd.0013663 2026-05-14T14:00:00Z 2026-05-14T14:00:00Z <p>by Paulo C. Manrique-Valverde, Chloe M. Hasund, Katrina A. Kelley, Jorge-Eduardo Amaya-Romero, Myriam Arévalo-Herrera, Raphael Brosula, Jose E. Calzada, Stella M. Chenet, Vladimir Corredor, Angela M. Early, Gustavo Fontecha, David A. Forero-Peña, Sócrates Herrera, Justin T. Lana, Margaret Laws, Reza Niles-Robin, Nicanor Obaldia, Ana M. Santamaria, Philipp Schwabl, Sarah Auburn, Daniel E. Neafsey</p> <i>Plasmodium vivax</i> is the main cause of malaria outside of sub-Saharan Africa, and in many settings it presents significant challenges to malaria elimination efforts. Despite some control successes in the Americas, regional annual case counts of malaria have increased by over 25% between 2014 and 2023, largely driven by <i>P. vivax.</i> Genomic surveillance can play a key role in understanding the extent to which disease persistence represents indigenous transmission as opposed to introduction of new strains through migration, and whether specific variants evade control measures. Efforts to make <i>P. vivax</i> genomic surveillance more cost-effective have led to the development of targeted sequencing-based methods, which strike a varying balance between assay sensitivity and breadth/informativeness. We introduce two new highly sensitive multiplexed amplicon sequencing panels for <i>P. vivax</i>: PvGTSeq and PvCRiSP. PvGTSeq requires selective whole-genome amplification (sWGA) and contains 249 amplicons—36 for antimalarial resistance and 213 for population structure—optimized for Latin America but applicable to all continents. PvCRiSP features four highly polymorphic amplicons that operate without sWGA and is designed to estimate complexity of infection (COI), identify instances of clonal transmission, and characterize recurrent episodes. Both panels use a single multiplex PCR with non-proprietary reagents, achieve ≥75% amplicon recovery at parasitemias as low as five parasites/μL, and PvCRiSP remains effective with low quality DNA. PvGTSeq showed high sequencing accuracy (error rate 3.85e-4% - 2.87e-3%), and both panels efficiently detected alleles from minority clones in simulated polyclonal infections. We validated both panels with samples from Colombia, Guyana, Honduras, Panama, and Venezuela, and performed in-silico assessments using data from 16 countries worldwide, confirming that these two panels have high power to discriminate samples and assign global geographic origin to imported cases. These panels will therefore be useful tools for <i>P. vivax</i> molecular surveillance in diverse geographic settings. Ruifang Song Yong Shen Fuying Guo Qing Zhen Shuo Kou Shun Zha Xiaojun Yu Zhuo Li Shanshan Song Jiaxin Hao Yiting Zhang Yingtong Wang Tian Ma Tiejun Shui Xiangyu Yan Weijia Zhao 10.1371/journal.pntd.0013209 2026-05-14T14:00:00Z 2026-05-14T14:00:00Z <p>by Ruifang Song, Yong Shen, Fuying Guo, Qing Zhen, Shuo Kou, Shun Zha, Xiaojun Yu, Zhuo Li, Shanshan Song, Jiaxin Hao, Yiting Zhang, Yingtong Wang, Tian Ma, Tiejun Shui, Xiangyu Yan, Weijia Zhao</p> Background <p>Leprosy remains a neglected tropical disease and major global public health challenge, particularly in developing countries with severe healthcare workforce shortages hindering control. The southwestern border, represented by Yunnan Province, is a core endemic area. A comprehensive assessment of its leprosy workforce is critical for achieving elimination.</p> Methods/Results <p>In October 2024, a cross-sectional census survey evaluated 423 Leprosy prevention and control personnel across all 129 counties in Yunnan Province, China. These counties were categorized into high (I), moderate (II), and low (III) endemicity areas based on their leprosy burden. Distinct workforce patterns emerged among 423 personnel, Category I areas exhibited the highest proportion of personnel aged 30–39 years (37.5%), along with the continuing education participation (78.5%), the highest full-time employment rate (52.5%), and strongest prescription protocol awareness (72.5%). Category II areas featured the oldest personnel profile (35.0% aged ≥50 years),with a moderate positive correlation between age and years of service (r = 0.58), highest continuing education participation (78.8%). In contrast, Category III areas had the highest proportion under 30 years (20.5%), and highest proportion of personnel with Bachelor’s degrees or higher (69.9%) lowest full-time employment rate (38.4%), highest compensation dissatisfaction (38.4% “below average”), and lowest intention to leave (8.2%). Pervasive workforce aging existed (at least 30% of personnel ≥50 years in Category II and III) and widespread technical gaps (>75% Category III areas lacked essential lab skills). Part-time staffing was common (47.5%-61.6% across categories). Despite over >90% of personnel in these three categories rated their compensation as “average or below”, compensation and career challenges were most acute in Categories I and II.</p> Conclusions <p>The Yunnan leprosy workforce shows strengths, notably high continuing education participation (78.5%) and balanced clinical/preventive staffing. However, it faces significant challenges, workforce aging, shortage of highly qualified personnel, and limited lab capacity. Urgent intervention measures are needed to revitalize the workforce, enhance training, and strategically allocate resources to expedite the achievement of leprosy elimination.</p> Lei Zhang Lilin Li Le Sun Danhong Cheng Shuwen Yang Yu Wang Xing Yan Xiaoyan Zhu Huan Zhang Cuiying Li Weixun Chunyu 10.1371/journal.pntd.0014340 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Lei Zhang, Lilin Li, Le Sun, Danhong Cheng, Shuwen Yang, Yu Wang, Xing Yan, Xiaoyan Zhu, Huan Zhang, Cuiying Li, Weixun Chunyu</p> Ulcerative colitis (UC) is a chronic inflammatory bowel illness with few treatment options, which means that new ways to treat it are needed. This study examined the protective effects and mechanisms of <i>Paragonimus proliferus</i> metacercaria-derived antigens (PmAg) in a dextran sulfate sodium (DSS)-induced mouse ulcerative colitis model. We discovered that intraperitoneal delivery of PmAg substantially mitigated colitis severity, as demonstrated by decreased weight loss, lower disease activity index scores, maintained colon length, and enhanced histopathological findings. Mechanistically, PmAg inhibited pro-inflammatory cytokines (IL-1β, TNF-α), increased the anti-inflammatory cytokine IL-10, and bolstered antioxidant defenses (SOD, GSH). It also restored the integrity of the intestinal barrier by boosting the number of goblet cells and the expression of tight junction proteins (Occludin, Claudin-1), while stopping the activation of the nuclear factor-kappa B (NF-κB) signaling pathway. Moreover, 16S rRNA sequencing demonstrated that PmAg reinstated gut microbiota α-diversity, diminished pathogenic genera (e.g., <i>Escherichia-Shigella</i>), and enhanced beneficial taxa (e.g., <i>Lachnospiraceae_NK4A136_group</i> and <i>Alistipes</i>). Integrated fecal metabolomics research revealed that PmAg altered metabolic profiles, specifically as significantly enriched the primary bile acid biosynthesis pathway, alpha-Linolenic acid metabolism pathway and Ubiquinone and other terpenoid-quinone biosynthesis pathways. In conclusion, our results suggested that PmAg could mitigates experimental colitis in mice by anti-inflammatory, improving gut microbiota and modulating fecal metabolomics. Yanfen Niu Yihui Zhang Song Zhou Guang Liu Yongming Chen Yuming Yuan Guoyi Du Jue Liu 10.1371/journal.pntd.0014337 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Yanfen Niu, Yihui Zhang, Song Zhou, Guang Liu, Yongming Chen, Yuming Yuan, Guoyi Du, Jue Liu</p> Background <p>Plague is a severe zoonotic disease caused by <i>Yersinia pestis</i>, a pathogen characterized by high infectivity and mortality rates. Historically, three global pandemics have inflicted heavy disasters on human society. Despite improvements in control measures and the application of antibiotics, plague has been somewhat controlled; however, since the beginning of the 21st century, plague outbreaks have continued to occur in regions with a high burden of neglected tropical diseases (NTDs) in Africa, the Americas, and Asia. In recent years, the rapid development of technologies such as molecular biology, immunology, and bioinformatics has propelled significant advancements in plague diagnostics, vaccine development, and transmission mechanisms. However, there has been a lack of systematic quantitative analysis of the distribution characteristics, evolving hotspots, and frontier trends of plague research, which makes it challenging to provide comprehensive scientific support for research and control decision-making.</p> Methods <p>Search for relevant literatures on plague that were published in the Web of Science Core Collection and PubMed database from January 1, 2016 to November 12, 2025.Bibliometric methods were adopted, and software including COOC 20.6, VOSviewer 1.6.20, and Anaconda were used to analyze the publication trend, distribution of institutions, national cooperation network, keyword co-occurrence clustering, and the annual variation trends.</p> Results <p>A total of 1994 documents were finally included. The annual number of publications showed an overall fluctuating upward trend, with a significant growth rate from 2020 to 2021 (annual growth rate of 10.44%). Core research institutions included the U. S. Centers for Disease Control and Prevention and Beijing Institute of Microbiology and Epidemiology. The United States, China, France and Madagascar were the main core countries for cooperation. Keyword co-occurrence clustering identified five major research fields, which were plague vaccine development and immune mechanism, ecology and vector control, historical epidemiology and public health, epidemiology and transmission chain, and plague-related infectious diseases and biosafety. The research trends analysis showed that from 2016 to 2020, the plague research mainly focused on keywords such as “Rodents” “Epidemiological Survey” “Human Plague” and “Fleas”. From 2021 to 2025, “Phylogenetic Analysis,” “Public Health,” and “Madagascar” newly entered the top 20 keyword list; the frequencies of “Black Death” and “Infectious Disease” increased significantly, while the frequencies of “Plague Vaccine” and “Prairie Dogs” remained relatively stable.</p> Conclusions <p>Over the past decade, remarkable achievements have been made in plague research. Interdisciplinary integration and technological innovation have continued to deepen. However, global collaboration remains insufficiently developed. In the future, it is necessary to foster broader cross-regional cooperation, accelerate the research, development and translation of vaccines and diagnostic technologies, integrate multiple technologies to construct a precise prevention and control system. and enhance the global collaborative prevention and control capabilities of plague.</p> Michael Odhiambo Israel Wuresah Geofrey Chepchieng Nikita Kubal Alejandro Krolewiecki Charles Mwandawiro Alan Brooks Collins Okoyo 10.1371/journal.pntd.0014317 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Michael Odhiambo, Israel Wuresah, Geofrey Chepchieng, Nikita Kubal, Alejandro Krolewiecki, Charles Mwandawiro, Alan Brooks, Collins Okoyo</p> Background <p>Soil-transmitted helminthiases (STH) infect over 1.5 billion people worldwide, causing serious health issues, especially among women of reproductive age and school-age children. Mass drug administration (MDA) using albendazole or mebendazole is the main method of control. However, high reinfection rates and limited integration with water, sanitation, and hygiene (WASH) and routine primary health care programs make the effectiveness of long-term control and elimination challenging.</p> Methods <p>This scoping review aimed to systematically map and summarize cost-effectiveness evidence for STH control and elimination efforts. Following Arksey and O’Malley’s framework and Preferred Reporting Items for Systematic reviews and Meta-Analysis extension for Scoping Reviews (PRISMA-ScR) guidelines, we identified 22 studies published from 1993 to 2025 through PubMed and Google Scholar.</p> Principal findings <p>Most studies focused on MDA intervention with school or community-based delivery platforms. School-based delivery costs ranged from $0.03 to $0.76 per child, and community-wide delivery costs ranged from $0.27 to $1.74 per individual. Community-wide programs were estimated to have greater impact and cost-effectiveness in high-prevalence areas, costing $28 to $198 per disability-adjusted life year averted. Programs that delivered multiple disease interventions (integrated programs) showed the highest economic returns, primarily due to shared delivery platforms and reduced operational costs. The main factors influencing cost-effectiveness included treatment coverage, baseline prevalence, and delivery costs. Evidence gaps for cost-effectiveness still exist for preschool-aged children and integration with WASH.</p> Conclusion <p>This scoping review showed that context affects the cost-effectiveness of STH intervention programs. Community-wide and integrated MDA strategies offered more economic value than school-based delivery in areas with high prevalence.</p> Rahmat Dapari Safiyeh Tayebi Ana Lorena Ruano Timothy C. Guetterman Seok Mui Wang Siti Hafizah AB Hamid Sohel Rahman Jürgen Pilz Nazri Che Dom Ubydul Haque 10.1371/journal.pntd.0014316 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Rahmat Dapari, Safiyeh Tayebi, Ana Lorena Ruano, Timothy C. Guetterman, Seok Mui Wang, Siti Hafizah AB Hamid, Sohel Rahman, Jürgen Pilz, Nazri Che Dom, Ubydul Haque</p> Victor O. Zorrilla Andrés Carrazco-Montalvo Liz J. Espada Leonardo Fárez-Noblecilla Marisa E. Lozano Michael Kosoy Clifton McKee Craig A. Stoops Ryan T. Larson Renato León Gissella M. Vásquez 10.1371/journal.pntd.0014288 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Victor O. Zorrilla, Andrés Carrazco-Montalvo, Liz J. Espada, Leonardo Fárez-Noblecilla, Marisa E. Lozano, Michael Kosoy, Clifton McKee, Craig A. Stoops, Ryan T. Larson, Renato León, Gissella M. Vásquez</p> Phlebotomine sand flies are blood-sucking dipterans widely distributed in tropical and subtropical areas of the Americas and are important vectors of leishmaniasis caused by <i>Leishmania</i> spp. parasites and Carrion’s disease caused by the bacteria <i>Bartonella bacilliformis</i>. Both are a significant economic burden in rural areas and a major risk to military personnel deployed to endemic areas. To better understand transmission of these pathogens and epidemiological trends, sand flies were collected from nine sites across the Ecuador-Peru border region in 2015 and 2017 and screened for <i>Leishmania</i> using PCR targeting kinetoplast DNA, and <i>Bartonella</i> using PCR targeting the 16S-23S internal transcribed spacer (ITS) region, citrate synthase (<i>gltA</i>) gene, and NADH dehydrogenase subunit G (<i>nuoG</i>) gene. A total of 548 sand flies belonging to 15 species and 2,711 sand flies belonging to 11 species were collected in Ecuador and Peru, respectively. <i>Pintomyia (Pifanomyia) robusta</i> was generally the most abundant species found across sites sampled in Ecuador and Peru. In the Chinchipe River basin, <i>Pi. (Pif.) robusta</i>, <i>Pintomyia (Pifanomyia) maranonensis</i>, and <i>Lutzomyia (Helcocyrtomyia) castanea</i> were collected on both sides in Zamora-Chinchipe, Ecuador, and Namballe, Peru. Of the 637 phlebotomine sand fly pools screened, no <i>Leishmania</i> positives were found; however, nine pools of <i>Pi. (Pif.) robusta</i> collected in Ecuador were positive for <i>B. bacilliformis</i> based on phylogenetic analysis of the <i>gltA</i> gene. One <i>Pi. (Pif.) maranonensis</i> from Peru and one <i>Pi. (Pif.) robusta</i> from Ecuador were positive for <i>Bartonella</i> DNA sequences that were close to <i>Candidatus</i> Bartonella rondoniensis based on <i>gltA</i>. This is the first reported detection of <i>B. bacilliformis</i> DNA in <i>Pi. (Pif.) robusta</i>, providing evidence for the role of this sand fly species in transmission of this pathogen at the Ecuador-Peru border. Erni Juwita Nelwan Yunita Windi Anggraini Sabighoh Zanjabila Budi Setiawan Suhartiningsih Suhartiningsih Farida Dwi Handayani Jeny Jeny Linda Erlina Fadilah Fadilah J. Kevin Baird Raph L. Hamers Suwarti Suwarti 10.1371/journal.pntd.0014243 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Erni Juwita Nelwan, Yunita Windi Anggraini, Sabighoh Zanjabila, Budi Setiawan, Suhartiningsih Suhartiningsih , Farida Dwi Handayani, Jeny Jeny , Linda Erlina, Fadilah Fadilah, J. Kevin Baird, Raph L. Hamers, Suwarti Suwarti</p> Recurring outbreaks of leptospirosis in flood-prone areas caused by heavy rainfall pose a major public health concern, particularly in megacities such as Jakarta, Indonesia. From December 2019 through February 2020, Jakarta experienced a large leptospirosis outbreak due to extensive flooding following extreme monsoonal rainfall. We conducted a comprehensive retrospective analysis of the outbreak based on complete surveillance data from all five districts and 42 of 44 subdistricts in Jakarta. A total of 282 cases (97 suspected, 153 probable, and 32 confirmed) were reported in West (n = 162), South (n = 64), East (n = 30), North (n = 14) and Central (n = 12) Jakarta. Cases were predominantly adult males exposed to floodwaters. Of 241 cases tested, 164 (68.0%) had a positive IgM-based rapid diagnostic test (RDT). Of 118 cases tested with TaqMan RT-PCR targeting <i>lipL32</i>, 32 (27.1%) were positive. Of 95 cases tested with both assays, the combined detection rate was 74.7% (71/95); of whom 27 were positive by both RDT and RT-PCR. RT-PCR identified 5 additional RDT-negative cases, all of whom had fever <7 days. We sequenced 42 archived blood samples using Multi Locus Sequence Typing (MLST) and identified <i>Leptospira interrogans</i> and <i>L. borgpeterseni</i> as the predominant species. The findings emphasise the importance of rapid and early laboratory-based diagnosis during leptospirosis outbreaks in flood-prone urban areas, to better target public health interventions. Climate-resilient urban planning is critical for vulnerable megacities in low-resource settings, where complex environmental and infrastructural challenges are compounded by the effects of a changing climate. Mischka Moodley Michalis Kostapanos Laura Iavarone Marguerite Bracher Silvia M. Lavezzi Maria Davy Pooja Singh Ramya Bhat Deborah Wong Hema Sharma Lionel K. Tan Timothy J. Miles 10.1371/journal.pntd.0014181 2026-05-13T14:00:00Z 2026-05-13T14:00:00Z <p>by Mischka Moodley, Michalis Kostapanos, Laura Iavarone, Marguerite Bracher, Silvia M. Lavezzi, Maria Davy, Pooja Singh, Ramya Bhat, Deborah Wong, Hema Sharma, Lionel K. Tan, Timothy J. Miles</p> Background <p>Visceral leishmaniasis (VL) is a neglected tropical disease caused by kinetoplastid parasites. If left untreated, VL has a case fatality >90%, making treatment essential for patient survival. Existing therapies have several limitations that impact treatment outcomes, including toxicity, requirement for parenteral administration, long treatment duration, and development of parasite resistance, driving the need for newer therapies. This Phase 1 first-time-in-human study evaluated the tolerability, safety, and pharmacokinetics (PK) of GSK3494245, a novel highly selective <i>Leishmania</i> kinetoplastid proteasome inhibitor.</p> Methodology <p>This was a randomized, double-blind, placebo-controlled, single ascending dose study evaluating the oral administration of GSK3494245 in healthy participants aged 18 to ≤55 years (NCT04504435). The study was conducted at a single center in the United Kingdom between October 2020 and January 2024.</p> Findings <p>GSK3494245 had an acceptable safety profile following administration of single doses of up to 240 mg in healthy male participants. All adverse events (AEs) were considered resolved or recovered, with most AEs being of mild or moderate severity. One serious AE of mild tachycardia was reported. The increase in exposure was slightly more than dose-proportional and approximately proportional under fasted and fed conditions, respectively. The median time to reach the maximum plasma concentration (C_max) ranged from 0.50 to 1.79 hours, and the geometric mean of the elimination half-life ranged from 1.04 to 2.27 hours. Two participants met the C_max stopping criterion.</p> Conclusion <p>GSK3494245 showed an acceptable safety profile over the dose range studied. However, based on its observed PK profile, GSK3494245 is unlikely to achieve the effective dose while ensuring safe exposure within the once- or twice-daily monotherapy target product profile recommended by the Drugs for Neglected Diseases initiative.</p> Clinical Trial Registration <p>NCT Number (www.clinicaltrials.gov): NCT04504435EudraCT, CTIS: 2019-004492-39</p> Abdul R. Anshad Muthuvel Atchaya Shanmugam Saravanan Amudhan Murugesan Siyana Fathima Ethihas R. Mahasamudram Rajendran Kannan Marie Larsson Esaki M. Shankar 10.1371/journal.pntd.0014327 2026-05-12T14:00:00Z 2026-05-12T14:00:00Z <p>by Abdul R. Anshad, Muthuvel Atchaya, Shanmugam Saravanan, Amudhan Murugesan, Siyana Fathima, Ethihas R. Mahasamudram, Rajendran Kannan, Marie Larsson, Esaki M. Shankar</p> Dengue virus (DENV) appears to manipulate several cellular metabolic pathways to permit its replication and immune evasion in the host. Here, we employed high-resolution mass spectrometry (HR-MS) to investigate the serum metabolomic landscape of clinical DENV infection. Serum specimens from primary dengue (n = 11), secondary dengue (n = 9) samples, and healthy controls (n = 10) were used for untargeted and targeted metabolomic quantification on a Waters Xevo G2-XS QTof Mass Spectrometer. The binding potential of selected ligands against DENV NS1, NS3, and NS5 was evaluated. Crystal structures were retrieved from Protein Data Bank and prepared using the Schrodinger’s protein preparation wizard. Based on findings from untargeted metabolomics, we validated certain bioactive lipid metabolites using commercial enzyme immunoassays. Serum metabolomic profiling revealed multiple distinct patterns for primary and secondary dengue versus controls. A consistent peak was observed at 2.06 mins across all samples. Certain bioactive lipid metabolites, such as, lysophospholipids, phosphatidylcholines, phosphatidylserines, and phosphatidylinositols, were detected alongside carnitine fragments, ceramides, diacylglycerols (DAGs), and bile acid conjugates in dengue. Molecular docking showed that DAG consistently exhibited strong binding to all the DENV proteins. Notably, LPC 22:6 showed a selectively strong affinity for NS5. Enzyme validation showed that in the secondary dengue cohort, LPC was significantly elevated than primary and healthy controls (p < 0.05). Our investigations of the metabolomic landscaping, unveiled certain characteristic anabolic shift revealing metabolic vulnerabilities in clinical DENV infection, warranting investigations for use as potential biomarkers of inflammation in disease diagnosis and prognosis. The PLOS Neglected Tropical Diseases Editors 10.1371/journal.pntd.0014311 2026-05-12T14:00:00Z 2026-05-12T14:00:00Z <p>by The PLOS Neglected Tropical Diseases Editors </p> Elias Jackson Veronique Dermauw Guy Eyaba Tchamdja Mohammad Shah Jalal Katrina Di Bacco Pierre Dorny Amanda Janitz Hélène Carabin 10.1371/journal.pntd.0014281 2026-05-12T14:00:00Z 2026-05-12T14:00:00Z <p>by Elias Jackson, Veronique Dermauw, Guy Eyaba Tchamdja, Mohammad Shah Jalal, Katrina Di Bacco, Pierre Dorny, Amanda Janitz, Hélène Carabin</p> Background <p><i>Taenia solium</i> cysticercosis/taeniasis disease complex is a multifaceted neglected tropical zoonosis. Previous reviews on <i>T. solium</i> cysticercosis/taeniasis risk factors have been limited by geographic region and/or host, making it difficult to understand the complex web of causes underlying infection at the various stages of its life cycle. Our objective was to elucidate the causal mechanisms involved in the transmission of <i>T. solium</i> to all hosts through a systematic review of the literature.</p> Methodology/Principal findings <p>We conducted a systematic literature search about the epidemiology of <i>T. solium</i> infection published before May 2020 (OpenScience protocol https://doi.org/10.17605/OSF.IO/U64K3). We searched PubMed, Web of Science, and CABAbstracts, and reference lists of systematic reviews identified in our searches for relevant studies meeting the inclusion criteria: i) describing a risk factor-outcome association related to <i>T. solium</i> infection, and ii) published in English, French, Spanish, or Portuguese. Qualitative or quantitative epidemiologic associations were extracted and evaluated according to a modified version of the Quality Assessment Tool for Quantitative Studies. Our review is reported according the PRISMA guidelines. We extracted 876 associations from 159 studies. Risk factors of human cysticercosis are well-studied, while taeniasis is least-studied (10% of associations). The evidence suggests that male gender is a risk factor for human cysticercosis while female sex is a risk factor for porcine cysticercosis. The direction of association did not differ according to the quality of the studies across most studied risk factors.</p> Conclusions/Significance <p>Our rigorous systematic review is the first to consider risk factors of human and porcine <i>T. solium</i> disease simultaneously, allowing a synthesis of information from both hosts in order to elucidate causal mechanisms for risk factors of cysticercosis. Our work also highlights areas of <i>T. solium</i> epidemiology which are understudied, such as the causal mechanisms of taeniasis.</p> Danilo de Carvalho-Leandro Francisco C. Ferreira Nadia Fernández Santos William J. Sames IV Yuexun Tian Martial L. Ndeffo-Mbah Erica A. Costa Michael J. Turell Gabriel L. Hamer Tereza Magalhaes 10.1371/journal.pntd.0013516 2026-05-12T14:00:00Z 2026-05-12T14:00:00Z <p>by Danilo de Carvalho-Leandro, Francisco C. Ferreira, Nadia Fernández Santos, William J. Sames IV, Yuexun Tian, Martial L. Ndeffo-Mbah, Erica A. Costa, Michael J. Turell, Gabriel L. Hamer, Tereza Magalhaes</p> Madariaga virus (MADV), widely distributed in Latin America, can cause severe disease in humans and equids, yet key aspects of its transmission cycle remain unclear. To identify mosquitoes that could act as vectors of MADV, we assessed the vector competence of <i>Aedes aegypti</i>, <i>Ae. albopictus</i>, <i>Ae. taeniorhynchus</i>, <i>Culex tarsalis</i>, <i>Cx. coronator</i>, and <i>Cx. quinquefasciatus</i>, following oral exposure to MADV isolated in Panama (all species) or Brazil (<i>Ae. taeniorhynchus</i> only). We also evaluated temporal infection dynamics of MADV from Panama in <i>Ae. aegypti</i> and <i>Ae. albopictus</i>. MADV RNA and infectious virus were quantified in mosquito bodies, legs, and saliva. At 14 days post-exposure, five of the six species tested had virus detected in all biological sample types, indicating their potential to become infected with and transmit MADV. Conversely, <i>Cx. quinquefasciatus</i> was susceptible to midgut infection and dissemination but had no positive saliva samples, suggesting limited transmission potential. <i>Aedes taeniorhynchus</i> showed higher infection probabilities with MADV from Brazil compared with MADV from Panama. Time-course analysis revealed distinct infection dynamics in <i>Ae. aegypti</i> and <i>Ae. albopictus</i>, with infection increasing over time in <i>Ae. aegypti</i>, but peaking at 7 days post-exposure and then gradually declining in <i>Ae. albopictus</i>. Our findings indicate that MADV may be compatible with multiple mosquito species with broad geographic distributions, reinforce the need to investigate species- and strain-specific mosquito-virus interactions and their influence on arbovirus transmission dynamics, and support a potential role for <i>Ae. taeniorhynchus</i> as an amplification and bridge vector in endemic regions. Ruchi Paroha Takeshi Saito Shintaro Yamada Christine Click Slobodan Paessler Junki Maruyama 10.1371/journal.pntd.0014326 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Ruchi Paroha, Takeshi Saito, Shintaro Yamada, Christine Click, Slobodan Paessler, Junki Maruyama</p> Lassa virus (LASV), the causative agent of Lassa fever (LF), poses a significant public health concern in endemic regions due to its high morbidity and mortality. Rapid and accurate diagnosis is critical for effective clinical management and containment during LF outbreaks. However, the genetic diversity of LASV, encompassing at least seven distinct lineages, poses a major challenge for the development of broadly reactive diagnostic tools. Therefore, there is an urgent need to develop detection methods that are effective across diverse lineages. To address this challenge, we generated a panel of cross-reactive monoclonal antibodies (mAbs) targeting the LASV nucleoprotein (NP). Several mAbs exhibited broad reactivity across LASV lineages I-VII in different immunoassays such as enzyme-linked immunosorbent assay (ELISA), western blotting, and immunofluorescence assay. Utilizing these broadly reactive mAbs, we developed an antigen-capture sandwich ELISA capable of detecting LASV NP from all seven lineages with high sensitivity. Our findings highlight a set of novel mAbs with broad cross-reactivity across lineage. In addition, we demonstrated their utility in a sandwich ELISA format for <i>pan</i>-LASV detection. This <i>pan</i>-LASV diagnostic approach offers a promising tool for improved LF diagnosis in both clinical and epidemiological settings. Luís Arthur Brasil Gadelha Farias Osvaldo Mariano Viana Neto Ednaldo Pereira Lima Sobrinho Caroline Lucena de Almeida Vale Geraldo Bezerra Silva Júnior Elizabeth de Francesco Daher Glaura Fernandes Teixeira de Alcântara Antônio Silva Lima Neto Tania Mara Silva Coelho Maura Salaroli de Oliveira Lauro Vieira Perdigão Neto 10.1371/journal.pntd.0014315 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Luís Arthur Brasil Gadelha Farias, Osvaldo Mariano Viana Neto, Ednaldo Pereira Lima Sobrinho, Caroline Lucena de Almeida Vale, Geraldo Bezerra Silva Júnior, Elizabeth de Francesco Daher, Glaura Fernandes Teixeira de Alcântara, Antônio Silva Lima Neto, Tania Mara Silva Coelho, Maura Salaroli de Oliveira, Lauro Vieira Perdigão Neto</p> Background <p>Leptospirosis is a globally distributed zoonotic disease with a broad clinical spectrum. Central nervous system (CNS) involvement is uncommon and under-recognized, and leptospiral-associated meningitis (LAM) is primarily described in isolated case reports and small series. No study to date has integrated institutional cases with published reports to characterize cerebrospinal fluid (CSF) profiles in LAM.</p> Methods <p>We conducted a retrospective cohort study of patients with meningitis, encephalitis, or meningoencephalitis admitted to a tertiary infectious diseases center in northeastern Brazil. Cases of LAM were identified among patients with aseptic meningitis. In parallel, a narrative literature review was performed, and a meta-summary of published cases was constructed. CSF parameters from both datasets were extracted and analyzed using descriptive statistics and graphical (boxplot) methods.</p> Results <p>Among 809 patients with meningitis or encephalitis, 447 presented with aseptic meningitis. Three cases (0.67%) of LAM were identified, presenting as isolated meningitis or meningoencephalitis. Among 179 patients, including 176 identified in the medical literature, mean CSF values were: cellularity 68 cells/mm³ (range, 30–7800), lymphocytes 73% (0–100), neutrophils 18% (0–96), glucose 60 mg/dL (0–140), and protein 67–90 mg/dL (31–2590). CSF findings in the institutional cohort showed mild to moderate pleocytosis with lymphocytic predominance, normal to elevated glucose levels, and increased protein concentrations. The integrated analysis of cohort and published cases—the first combined CSF profile synthesis of LAM—demonstrated a consistent pattern of lymphocytic pleocytosis, mildly elevated cellularity, preserved glucose levels, and increased protein, with substantial inter-case variability.</p> Conclusion <p>LAM is an uncommon but clinically relevant cause of CNS infection in endemic settings. This study provides the first integrated synthesis of CSF profiles from both institutional cases and published literature, supporting a characteristic but variable CSF pattern in leptospiral CNS disease. Clinicians should consider leptospirosis in patients with aseptic meningitis in endemic areas, particularly when epidemiological risk factors are present. Improved diagnostic capacity and prospective studies using standardized criteria are needed to better define disease burden and refine diagnostic and therapeutic approaches.</p> Angelika Proskurnicka Karolina Duk Tomasz Hutsch Grażyna Hoser Robert Wrzesień Tomasz Skirecki Jacek Bielecki Tomasz Jagielski 10.1371/journal.pntd.0014312 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Angelika Proskurnicka, Karolina Duk, Tomasz Hutsch, Grażyna Hoser, Robert Wrzesień, Tomasz Skirecki, Jacek Bielecki, Tomasz Jagielski</p> <i>Prototheca</i> spp. are unicellular, yeast-like microalgae, and the only plant lineage known to cause opportunistic infections in vertebrates, including humans. The aim of the study was to establish a comprehensive murine model of protothecosis to systematically investigate the influence of <i>Prototheca</i> species, inoculum dose, infection route, and host immune status on disease development and severity. Three pathogenic (<i>P. wickerhamii</i>, <i>P. bovis</i>, <i>P. ciferrii</i>) and one saprophytic (<i>P. stagnora</i>) species were used to infect immunocompetent or athymic mice. A total of 324 animals were split into 54 groups according to inoculum size (10⁶ or 10⁷) and infection route (subcutaneous, intramammary, or intraperitoneal). Six weeks post-infection, mice were euthanized, and organs were collected for microbiological and histopathological analyses. All four <i>Prototheca</i> species produced infection in mice, yet the infection potential differed considerably between the species. <i>P. ciferrii</i> exhibited the highest infection rate (61.1%), followed by <i>P. bovis</i> (45.8%) and <i>P. wickerhamii</i> (31.9%), whereas <i>P. stagnora</i> was the least virulent (11.1%). Athymic mice were markedly more susceptible compared to wt mice (45.1% <i>vs</i>. 29.9%) and more prone to develop multifocal infections. Higher inocula (10⁷) increased infection yield, while the inoculation route influenced the infection site but not its severity. In the cytokine profile, IL-10 and TNF-α were most prominently elevated, with significantly higher levels in wt than in athymic mice. This study highlights three hallmarks of protothecal disease: a species-specific infection pattern, chronic, asymptomatic infection as the clinical manifestation, and the essential role of host immunity in determining disease trajectory and severity. Lilia Zribi Maria Paola Maurelli Mariele De Santi Maria Ortensia Montella Aida Bouratbine Valentina Foglia Manzillo Laura Rinaldi Karim Aoun Gaetano Oliva 10.1371/journal.pntd.0014299 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Lilia Zribi, Maria Paola Maurelli, Mariele De Santi, Maria Ortensia Montella, Aida Bouratbine, Valentina Foglia Manzillo, Laura Rinaldi, Karim Aoun, Gaetano Oliva</p> Background <p>Tunisia represents the perfect example of a Mediterranean Country where different <i>Leishmania</i> species may express their infectivity causing Visceral leishmaniasis by <i>Leishmania</i> (<i>L</i>.) <i>infantum</i>, Zoonotic Cutaneous Leishmaniasis by <i>L</i>. <i>major</i> and chronic cutaneous leishmaniasis by <i>L. tropica</i>. The recent detection of <i>L. major</i> in two dogs living in Tunisia confirms how this animal may host both visceral and cutaneous <i>Leishmania</i> species. The present study reports the results of 4 field surveys performed in central and southern districts of Tunisia: Zaghouan (ZA); Kairouan (2 surveys, K1 and K2); Tataouine (TA), to assess the prevalence of <i>Leishmania</i> species in dog.</p> Methods <p>One hundred and sixteen dogs were enrolled. Blood, lymph node and skin samples were collected with theowner’ consent. Thirty-two were enrolled during 2021 in ZA (n = 32), fifty-four were enrolled in KA during 2022 (K1; n = 22) and 2024 (K2, n = 32) and thirty were enrolled in TA during 2024 (n = 30). All dogs correspond to new surveys other than those investigated in a previous study. In total 218 biological samples were analyzed by qPCR (kDNA), end-point PCR (ITS-1) and nested-PCR (SSUrRNA). The purified positive PCR products were sequenced. All dogs were classified as asymptomatic or with mild clinical signs, not specifically attributable to Canine Leishmaniosis due to the presence of fleas and tick infestation.</p> Results <p>Thirty-eight dogs tested positive by molecular techniques (32.75%%). <i>Leishmania infantum</i> was the most identified species (31/116, 26.72%). Extremely high prevalence was found in K2 (23/32, 71.87%) compared with the previous study K1 (10/22, 45.45%). One dog (K1) was positive to <i>L. tropica,</i> the first detection of this species in Tunisia, while two dogs (ZA and K2) confirmed the presence of <i>L. major</i>. Interestingly, we found for the first time a dog positive to <i>L. infantum/donovani</i> in TA, an arid area where no VL cases have been previously recorded.</p> Conclusions <p>This study pointed out the high circulation of <i>L. infantum</i> in north and central Tunisia and underlines as the dog can host all the 3 <i>Leishmania</i> species present in this country.</p> Yanan Cai Yamei Wei Guoyi Du Xinyang Zhang Zhenkun Wang Zhengguang Wang Zhanying Han Yanbo Zhang Yonggang Xu Xu Han Jiandong Li Qi Li 10.1371/journal.pntd.0014250 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Yanan Cai, Yamei Wei, Guoyi Du, Xinyang Zhang, Zhenkun Wang, Zhengguang Wang, Zhanying Han, Yanbo Zhang, Yonggang Xu, Xu Han, Jiandong Li, Qi Li</p> Orthohantavirus infections pose a significant threat to human health, while numerous orthohantaviruses have been identified, suspected viral infections remain undiagnosed in the world, which highlights the need for further identification and characterization of viruses circulating in humans and host animals. In this study, viral metagenomics was utilized to investigate orthohantaviruses present in tissue samples collected from rodents trapped at the Bashang Grassland of Hebei Province, China. A total of 145 wild rodents belonging to six species were captured in the study area, and 725 tissue samples (lung, liver, kidney, spleen, gut) were collected in 2024. A Puumala orthohantavirus (PUUV), named Guyuan strain, was identified in <i>Myodes rufocanus</i>, with a positive rate of 0.69%. The complete genomic sequences of the L, M, and S segments were obtained and confirmed by Sanger sequencing. Phylogenetic analysis of these genomic sequences with those of other orthohantavirus species showed that the L, M, and S segments clustered with PUUV genomic sequences, while sharing a nucleotide sequence similarity of 81.2%, 80.2%, and 84.3% with previously characterized reference viral strains Kitahiyama128L, Tobetsu_04, and Baltic/205 Cg, respectively. Amino acid homology analysis demonstrated that the sequences exhibited the highest identity to PUUV Hokkaido strain at a level of 95.4%, 94.6%, and 97.0% respectively. Viral particles were observed in lung and kidney tissues using transmission electron microscopy, and viral protein antigen was detected in viral RNA-positive lung, liver, and kidney tissues through immunofluorescence assay with antibodies against the PUUV nucleocapsid protein, thereby confirming the virus’s multiorgan tropism. The results demonstrated that a distinct genotype of PUUV was circulating in rodents in the study areas, which may have implications for zoonotic transmission surveillance and public health management in Hebei Province. Deborah Pratt Yaw Awuku Larbi Magdalene Ofori Bright Agbodzi Jessica Wiley Ama O. A. Mante Selassie Kumordjie Miriam Eshun Stella Bour Nancy Enimil Juliana N. D. Acquah-Amaning Prince Ketorwoley Maame S. Boapea David D. Nutakor Musah Salisu Gertrude Stephens Emmanuel A. Boateng Tracy Adjandeh Joel Koomson Dennis Laryea Franklin Asiedu-Bekoe Patrick Kuma-Aboagye Patrick Mawupemor Avevor Argata G. Guracha Sally-Ann Ohene Terrel Sanders Michael Wiley Joseph Humphrey Kofi Bonney 10.1371/journal.pntd.0014248 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Deborah Pratt, Yaw Awuku Larbi, Magdalene Ofori, Bright Agbodzi, Jessica Wiley, Ama O. A. Mante, Selassie Kumordjie, Miriam Eshun, Stella Bour, Nancy Enimil, Juliana N. D. Acquah-Amaning, Prince Ketorwoley, Maame S. Boapea, David D. Nutakor, Musah Salisu, Gertrude Stephens, Emmanuel A. Boateng, Tracy Adjandeh, Joel Koomson, Dennis Laryea, Franklin Asiedu-Bekoe, Patrick Kuma-Aboagye, Patrick Mawupemor Avevor, Argata G. Guracha, Sally-Ann Ohene, Terrel Sanders, Michael Wiley, Joseph Humphrey Kofi Bonney</p> Background <p>Dengue, a mosquito-borne virus continues to be a public health concern in the tropics and subtropical parts of Africa. Due to the increase of urbanization, climate change and trans-Atlantic trade, the transmitting vector, Aedes aegypti, has become prevalent hence the rapid growth of the disease, globally. In Ghana, there have been sporadic laboratory-confirmed cases of dengue reported over the years through surveillance activities. However, despite the detection of these cases, Ghana had never experienced a major outbreak (unlike its neighboring countries) until July 2024.</p> Methods <p>During this outbreak, a total of 1471 suspected dengue fever specimen received from various health facilities in Ghana in NMIMR for molecular diagnostic testing using a RT-qPCR assay for dengue, chikungunya and Zika viruses and selected positives sequenced using Illumina Next Generation Sequencing.</p> Results <p>Dengue fever virus RNA was detected from 206 samples and serotyped as DENV-1 with one DENV-3 coinfection. Thirty-nine genomes were successfully generated after sequencing and phylogenetic analysis of DENV-1 strains revealed two main clusters with AFI isolates in Ghana and isolates from other West African countries (Côte d’Ivoire, Burkina Faso, Benin, and Senegal) circulating between 2017–2019. In 2023, DENV-1 was frequently isolated which could account for it being the predominant serotype transmitted in the recent outbreak.</p> Conclusion <p>The outbreak response and the case management procedures deployed by the health authorities during this outbreak were swift and was enough to prevent a fatal difficult-to-control situation. With the absence of a widely accepted commercialized vaccine and treatment for dengue fever, there is a need to enhance surveillance activities and control the vectors which can transmit DENV in-country to curb the occurrence of outbreaks.</p> Livia Mancinelli Marika Guercio Gianluca Vrenna Manuela Onori Claudio De Liberato Adele Magliano Andrea Diociaiuti Massimiliano Raponi Carlo Federico Perno May El Hachem Paola Bernaschi 10.1371/journal.pntd.0013535 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Livia Mancinelli, Marika Guercio, Gianluca Vrenna, Manuela Onori, Claudio De Liberato, Adele Magliano, Andrea Diociaiuti, Massimiliano Raponi, Carlo Federico Perno, May El Hachem, Paola Bernaschi</p> <i>Cordylobia anthropophaga,</i> commonly known as the <i>tumbu</i> fly, is a leading cause of cutaneous myiasis in sub-Saharan Africa. This condition is characterized by a papulopustular lesion that evolves in a painful boil-like nodule with central ulceration. Human infestations typically occur on covered body parts, due to the fly’s habit of laying eggs on damp clothing. This report describes the first cluster of three pediatric cases of furuncular myiasis caused by <i>C. anthropophaga</i>, diagnosed at the Bambino Gesù Children’s Hospital (Rome, Italy) between April 2024 and April 2025. All patients had recent travel history to endemic African regions and presented with cutaneous nodules, some accompanied by systemic signs such as fever or regional lymphadenopathy. Diagnosis was confirmed through clinical evaluation and morphological identification of the extracted larvae. Treatment consisted of occlusive therapy to facilitate larval expulsion or surgical extraction of the maggots and systemic antibiotics. One patient also exhibited a family cluster, with larval extraction from a parent. Laboratory results were largely unremarkable, consistent with localized infection. These cases highlight the growing relevance of imported myiasis in non-endemic countries like Italy, due to increased international travel and potential climate-driven changes in vector distribution. Enhanced awareness among travelers and healthcare providers, together with proactive public health measures, is crucial. By documenting these cases, we hope to contribute to the understanding of emerging parasitic diseases in non-endemic regions and reinforce the need for vigilance in this time of global environmental changes. Shomik Maruf Md Rasel Uddin Farhana Rahman Luba Soumik Kha Sagar Debashis Ghosh Md Sakhawat Hossain Megha Raj Banjara Axel Kroeger Christine Halleux Abraham Aseffa Dinesh Mondal 10.1371/journal.pntd.0013504 2026-05-11T14:00:00Z 2026-05-11T14:00:00Z <p>by Shomik Maruf, Md Rasel Uddin, Farhana Rahman Luba, Soumik Kha Sagar, Debashis Ghosh, Md Sakhawat Hossain, Megha Raj Banjara, Axel Kroeger, Christine Halleux, Abraham Aseffa, Dinesh Mondal</p> Background <p>Bangladesh became the first country to achieve World Health Organization (WHO) validation for eliminating visceral leishmaniasis (VL, Kala-azar) as a public-health problem in 2023. Sustaining this milestone demands a post-validation surveillance strategy that concentrates its efforts on residual transmission foci and deploys resources efficiently. We therefore conducted the country’s first Mouza-level micro-stratification to refine risk maps and guide targeted interventions.</p> Methods <p>The study used routinely reported VL line-list data (January 2017 – June 2025) from the national DHIS2 platform for every Upazila (sub-districts) that recorded ≥1 VL case. Each Mouza—the smallest administrative unit for land records comprising of one or more villages—was categorised as high (≥3 new VL cases), moderate (2 cases), low (1 case) or non-endemic (0 cases) over the nine-year period. Hot-spot maps were created in Python. Associations between endemicity (endemic vs non-endemic) were tested with chi-squared statistics, yielding odds ratios (OR) with 95% confidence intervals (CI).,</p> Results <p>Among 17,123 Mouzas in 128 case-reporting Upazilas, only 478 (2.8%) reported ≥1 new VL case between 2017 and 2025. High-endemic Mouzas (n = 33; 0.2%) accounted for 35% of total incident cases and clustered primarily in Mymensingh, Dhaka and Rajshahi divisions. However, year-on-year mapping showed reduction in the number of endemic Mouzas with no sustained new foci.</p> Conclusions <p>VL transmission in Bangladesh is now intensely focal, confined to <3% of Mouzas within historically endemic Upazilas. While broad surveillance coverage remains essential, micro-stratification at the Mouza level can guide the prioritization of targeted interventions, such as indoor residual spraying and active case detection in high-risk areas, improving program efficiency without compromising case detection. Periodic updating of Mouza-level risk maps will be essential to identify emerging hotspots, prevent resurgence, and inform strategies in other countries approaching VL elimination.</p> Larissa de Carvalho Medrado Vasconcelos Micaela Soledad Ossowski Edimilson Domingos Silva Daniel Dias Sampaio Tycha Bianca Sabaini Pavan Randrin Queiroz Viana Ferreira Felipe Silva Santos de Jesus Raúl Chadi Marisa Liliana Fernández Yolanda Hernández Karina Andrea Gómez Fred Luciano Neves Santos 10.1371/journal.pntd.0014310 2026-05-08T14:00:00Z 2026-05-08T14:00:00Z <p>by Larissa de Carvalho Medrado Vasconcelos, Micaela Soledad Ossowski, Edimilson Domingos Silva, Daniel Dias Sampaio, Tycha Bianca Sabaini Pavan, Randrin Queiroz Viana Ferreira, Felipe Silva Santos de Jesus, Raúl Chadi, Marisa Liliana Fernández, Yolanda Hernández, Karina Andrea Gómez, Fred Luciano Neves Santos</p> Chagas disease (CD) remains a major public health challenge in Latin America, largely due to persistent underdiagnosis, particularly in endemic and resource-limited settings. Rapid diagnostic tests (RDTs) offer a pragmatic approach to expand serological screening; however, their performance varies across epidemiological contexts and parasite genetic backgrounds. The TR Chagas Bio-Manguinhos test, based on recombinant chimeric antigens, has shown high sensitivity in previous evaluations, but data from Southern Cone remain limited. We conducted a cross-sectional diagnostic accuracy study using 311 anonymized human plasma samples obtained in Argentina, including 233 <i>Trypanosoma cruzi</i>–positive and 78 negative samples classified by a composite reference standard based on at least two independent serological assays. Diagnostic performance of the TR Chagas Bio-Manguinhos rapid test was assessed in terms of sensitivity, specificity, accuracy, likelihood ratios, diagnostic odds ratio, and agreement. Sensitivity was further stratified by country of origin (Argentina, Bolivia, Paraguay) and by cardiac involvement according to the Kuschnir classification. The TR Chagas Bio-Manguinhos test demonstrated high overall performance, with a sensitivity of 97.0% (95% CI: 93.9–98.5%), specificity of 94.9% (95% CI: 87.5–98.0%), and accuracy of 96.5% (95% CI: 93.8–98.0%). Agreement with the reference standard was almost perfect (κ = 0.91). Sensitivity remained consistently high across geographical origins and Kuschnir cardiac stages, with no statistically significant differences between subgroups. The negative likelihood ratio (0.03) indicated strong ability to rule out infection. These findings provide the first independent evidence supporting the high performance of the TR Chagas Bio-Manguinhos test in people from Argentina, Bolivia and Paraguay. The combination of very high sensitivity and robust specificity supports its use as a frontline serological screening tool for chronic <i>T. cruzi</i> infection in the Southern Cone, provided that reactive results are subsequently confirmed by laboratory- based assays in accordance with international guidelines. Juan David Ramírez Sarah M. Gunter Megan Coffee Norman Beatty Dawn M. Wetzel 10.1371/journal.pntd.0014297 2026-05-08T14:00:00Z 2026-05-08T14:00:00Z <p>by Juan David Ramírez, Sarah M. Gunter, Megan Coffee, Norman Beatty, Dawn M. Wetzel</p> Cutaneous leishmaniasis (CL), once considered a travel-associated tropical disease, is increasingly transmitted within the United States, particularly in southern regions. Despite mounting evidence of local transmission, public health recognition and preventive infrastructure remain limited. This Viewpoint highlights the urgent need to shift the U.S. CL response from questioning endemicity to preventing transmission. We review ecological, clinical, and surveillance data demonstrating the presence of competent vectors, animal reservoirs, and autochthonous human cases. Diagnostic delays, underreporting, and insufficient provider training contribute to missed prevention opportunities. Climate change and peri-urban rodent-human contact data further heighten future risk. A coordinated response is essential, including national notifiability, expanded diagnostics, integrated vector and reservoir surveillance, clinical education, and One Health–focused research. Without immediate action, CL risks becoming an entrenched, neglected zoonosis in the United States. Dieudonne Hakizimana Ladislas Nshimiyimana Jeanne Uwizeyimana Jean Bosco Mbonigaba Aimable Mbituyumuremyi Nathan Hitiyaremye Albert Tuyishime Alison Ower Karen Palacio Tonya Huston Ivy Sempele Eugene Ruberanziza 10.1371/journal.pntd.0014267 2026-05-07T14:00:00Z 2026-05-07T14:00:00Z <p>by Dieudonne Hakizimana, Ladislas Nshimiyimana, Jeanne Uwizeyimana, Jean Bosco Mbonigaba, Aimable Mbituyumuremyi, Nathan Hitiyaremye, Albert Tuyishime, Alison Ower, Karen Palacio, Tonya Huston, Ivy Sempele, Eugene Ruberanziza</p> Background <p>In Rwanda, about 1 in 3 people are affected by soil-transmitted helminthiases (38.7%) and 1 in 4 by schistosomiasis in high-risk areas (27.2% by point-of-care circulating cathodic antigen), both associated with poor water, sanitation, and hygiene (WASH). Mass drug administration (MDA) is the primary control strategy, but reported coverage may not reflect true reach. This study assessed MDA coverage of albendazole/mebendazole and praziquantel in selected districts, identified reasons for non-reach, examined factors associated with uptake, and described household WASH conditions.</p> Methods <p>A cross-sectional, community-based survey was conducted in five purposively selected districts using a stratified cluster design. Survey-weighted estimates summarized treatment reach (offered the drug), uptake (swallowed the drug), and household WASH conditions. Survey-weighted logistic regression was used to identify individual and implementation factors associated with uptake.</p> Results <p>Albendazole/mebendazole uptake was 91.9% (95% CI: 84.3–96.0), closely matching reported coverage of 96%. The main reasons for non-reach were drug stockouts (23.7%), absence during MDA (15.8%), and unwillingness to take the drug (15.2%). Praziquantel uptake was 88.0% (95% CI: 78.7–93.6), consistent with reported coverage of 80%. For praziquantel, unwillingness to take tablets (29.6%) and absence during MDA (14.4%) were the most common reasons for non-reach.Receiving sufficient information about MDA to make an informed decision was associated with higher odds of uptake for both ALB/MBZ (adjusted odds ratios [aOR]: 5.17, 95% CI: 2.01–13.27) and PZQ (aOR: 3.58, 95% CI: 1.33–9.64). For ALB/MBZ, finding MDA participation easy (aOR: 11.41, 95% CI: 2.59–50.16), feeling comfortable with the MDA distributor (aOR: 10.05, 95% CI: 2.43–41.47), and feeling comfortable with the MDA location (aOR: 3.23, 95% CI: 1.25–8.39) were each independently associated with higher uptake. For PZQ, males had significantly higher odds of uptake compared to females (aOR: 2.64, 95% CI: 1.15–6.07).Among households, 65.6% used improved drinking water sources, 91.4% obtained water from public places, 50.7% treated their water, 84.4% had improved toilets, 51.6% had visibly clean toilets, and 62% lacked a handwashing station.</p> Conclusion <p>MDA coverage in Rwanda exceeded WHO targets and closely matched reported estimates, reflecting strong implementation. Addressing remaining gaps in drug supply, MDA communication, and WASH infrastructure will be important to sustain and strengthen control of soil-transmitted helminthiases and schistosomiasis.</p> Lin Lin Ana Valeria Solano Fabiola Gonzales Mary Cruz Torrico Daniel Illanes Nuria Díez David Bermejo-Peláez Elena Dacal Ramón Vallés-López Lucia Pastor Roberto Mancebo-Martín María Jesús Ledesma-Carbayo Miguel Luengo-Oroz Jose M. Rubio Maria Flores-Chavez 10.1371/journal.pntd.0012955 2026-05-07T14:00:00Z 2026-05-07T14:00:00Z <p>by Lin Lin, Ana Valeria Solano, Fabiola Gonzales, Mary Cruz Torrico, Daniel Illanes, Nuria Díez, David Bermejo-Peláez, Elena Dacal, Ramón Vallés-López, Lucia Pastor, Roberto Mancebo-Martín, María Jesús Ledesma-Carbayo, Miguel Luengo-Oroz, Jose M. Rubio, Maria Flores-Chavez</p> Chagas disease affects 6–7 million people worldwide and causes approximately 12,000 deaths annually. Diagnostic methods vary by disease stage, with serological tests commonly used in the chronic phase, while microscopy and molecular techniques like PCR and LAMP are employed in the acute phase. While microscopy remains the most accessible tool in resource constrained settings, its effectiveness depends on skilled personnel, creating diagnostic bottlenecks. To overcome these limitations, we developed a portable, smartphone-integrated AI system for real-time <i>Trypanosoma cruzi</i> detection in microscopy images. The platform combines a 3D-printed microscope adapter which aligns the smartphone camera with the microscope ocular to digitize images, with telemedicine-enabled annotation workflows, and lightweight AI models (SSD-MobileNetV2, YOLOv8) deployed on smartphone for real-time analysis. Trained on a diverse dataset of human samples (478 images from 20 samples), including thick/thin blood smears and cerebrospinal fluid) and murine thin smears (570 images from 33 samples), the SSD-MobileNetV2 model achieved 86% precision, 87% recall, and 86.5% F1-score on human samples, demonstrating robust performance across variable imaging conditions. We additionally piloted a real-world experiment with the proposed system. Three thin blood smears were scanned by a user operating the smartphone-based system, with predictions generated in real time. Model outputs were benchmarked against expert annotations as the ground truth. At the object level, the algorithm achieved a precision of 67.1%, a recall of 96.4%, and an F1-score of 79.1%, showing high sensitivity under operational conditions with a configuration possibly suitable for screening. This system could enable rapid, accurate parasite detection in field settings without advanced infrastructure, addressing critical gaps in early diagnosis and monitoring. Its modular design allows adaptation to other pathogens and cellular structures, offering a scalable solution for neglected tropical disease diagnostics. By bridging AI innovation with microscopy, this approach holds promise for advancing equitable healthcare delivery in endemic regions and aligning with global health priorities. Sara Patiño-Gómez Luisa F. Naranjo-Vargas Daniel C. Aguirre-Acevedo Néstor Jaime Aguirre-Ramírez Elsio A. Wunder Jr Felipe de Oliveira Samanta C. das Chagas-Xavier Juan C. Quintero-Vélez 10.1371/journal.pntd.0014231 2026-05-06T14:00:00Z 2026-05-06T14:00:00Z <p>by Sara Patiño-Gómez, Luisa F. Naranjo-Vargas, Daniel C. Aguirre-Acevedo, Néstor Jaime Aguirre-Ramírez, Elsio A. Wunder Jr, Felipe de Oliveira, Samanta C. das Chagas-Xavier, Juan C. Quintero-Vélez</p> Background <p><i>Leptospira</i> are zoonotic agents with a complex transmission cycle that affects low-income and impoverished populations and causes significant economic losses in livestock.</p> Objective <p>To evaluate the interaction between people, animals, and the environment related to <i>Leptospira</i> infection in bovine farms in Urabá, Antioquia.</p> Methods <p>An exploratory cross-sectional study was conducted on cattle farms in Urabá, Antioquia. The proportion of pathogenic <i>Leptospira</i> infection in bovine and canine urine and environmental contamination in water and soil samples was estimated using molecular assays. Additionally, <i>Leptospira</i> seropositivity in humans, cattle, and canines was determined using the microagglutination test (MAT). Evaluation of composition characteristics of landscape was done and potential flooding areas were estimated. The domestic animals and human populations were characterized through descriptive analysis using productive and reproductive data and sociodemographic information, respectively. Then, associations between seropositivity/infection, antibody titers, <i>Leptospira</i> serogroups/species in cattle, canines, and humans, and productive, farms and landscape variables we explored using a mixed-data factor analysis.</p> Results <p>The proportion of seropositivity in cattle wa 76.9% (380/494). The most frequent serogroups on MAT were Mini, Tarassovi, Ballum, and Sejroe. In addition, molecular analysis indicated an infection rate of 4.0% (20/494) of the species <i>L. borgpetersenii</i> in cattle<i>.</i> Seropositivity in humans was 4.1% (3/73), with serogroups Icterohaemorrhagiae, Autumnalis, and Sejroe. Thirty-three percent (5/15) of dogs were seropositive for serogroups Canicola, Icterohaemorrhagiae, Ballum, and Autumnalis. The presence of <i>L. tipperaryensis</i> was detected in water and species <i>L. weilii</i> and <i>L. cinconiae</i> in soil. Evidence of high exposure to Leptospira was found in cattle. An association was also found between the serogroups circulating in humans and dogs (Autumnalis) and in humans and cattle (Sejroe), as well as forest fragmentation.</p> Conclusions <p>The importance of addressing the epidemiology of <i>Leptospira</i> infection from a comprehensive One Health approach is highlighted.</p> Wendemagegn Enbiale Borna Nyaoke Alemayehu Bekele Kedir Ahmed Mohammed Dereje Bedane 10.1371/journal.pntd.0013886 2026-05-06T14:00:00Z 2026-05-06T14:00:00Z <p>by Wendemagegn Enbiale, Borna Nyaoke, Alemayehu Bekele, Kedir Ahmed Mohammed, Dereje Bedane</p> Background Mycetoma is a chronic, progressively destructive infection that can result in severe disability and limb loss. In Ethiopia, diagnostic capacity and access to effective treatment remain limited, and the burden of mycetoma is poorly characterized. Recent clinical observations from the Afar Region suggest a high frequency of advanced disease and amputation, yet systematic evidence on the burden is lacking. This study aimed to describe the clinical burden of mycetoma, diagnostic and treatment practices, care-seeking patterns, and the extent of limb amputation at Dubti General Hospital in Afar region, northeastern Ethiopia. We conducted a facility-based retrospective review of all patients with a clinical diagnosis of mycetoma managed at Dubti General Hospital between September 2020 and August 2025. Demographic characteristics, clinical presentation, diagnostic investigations, treatment modalities, disability status, and surgical outcomes were summarized descriptively. Factors associated with delayed presentation (>12 months from symptom onset) were assessed using multivariable logistic regression. A total of 143 patients were identified, with a mean age of 30.9 years (SD ± 11.7); 79% were male, 85.3% resided in rural areas, and 46% were pastoralists. All cases involved the lower limb and presented with localized swelling. Pain (90.9%), warmth (54.5%), sinus formation (42.7%), and discharge (40.6%) were common. Diagnosis relied primarily on clinical assessment alone (58.7%), with limited use of imaging and biopsy. The mean duration of illness before first presentation was 33.8 months (SD ± 29), and 89.5% of patients presented after more than 12 months of symptoms. Compared with farmers, merchants had lower odds of delayed presentation (AOR = 0.89, 95% CI: 0.27–0.59). Nineteen patients (13.3%) underwent limb amputation, accounting for 23.5% of all orthopaedic amputations performed at the hospital during the study period. Disability at presentation was frequent, with 14.0% of patients experiencing severe motor impairment. Mycetoma in Afar predominantly affects young rural men and presents almost exclusively with advanced lower-limb disease. Profound diagnostic limitations, delayed care-seeking, and restricted surgical options contribute to poor outcomes. Integrating mycetoma into national neglected tropical disease strategies, strengthening early detection and diagnostic services, ensuring consistent access to essential medications, and expanding limb-sparing surgical capacity are critical to reducing preventable disability and aligning Ethiopia’s response with global NTD control targets.