PLoS Neglected Tropical Diseases最新文献

{"title":"Nuclear proteome analysis suggests spatial and temporal compartmentalization of metabolic enzymes during Trypanosoma cruzi cell cycle.","authors":"Ana Paula Menezes, Camila Gachet-Castro, Suzanne McGill, Richard Burchmore, Ariel Silber, Maria Carolina Elias, Júlia P C da Cunha","doi":"10.1371/journal.pntd.0013299","DOIUrl":"10.1371/journal.pntd.0013299","url":null,"abstract":"<p><p>Trypanosoma cruzi, the etiological agent of Chagas Disease (CD), represents a significant public health concern and serves as a valuable model for investigating the cell cycle in early-diverging eukaryotes. Its unique cellular features,including the absence of chromosome condensation and the coordination of nuclear division with specialized organelles, provide insights into non-canonical regulatory mechanisms, and underscore the pivotal role of the nucleus in maintaining genomic integrity and regulating fundamental processes such as DNA replication and gene expression. To investigate nuclear dynamics throughout the cell cycle, we synchronized T. cruzi epimastigotes at the G1/S transition using hydroxyurea (HU), followed their progression through S and G2/M phases, and quantitatively analyzed isolated nuclear proteins by LC-MS/MS. We identified 2,937 nuclear-enriched proteins, revealing distinct, phase-specific expression patterns. The G1/S phase was marked by increased levels of metabolic enzymes including those related to energy and nucleotide/nucleoside metabolisms. The S phase showed elevated abundance of canonical and variant histones, consistent with chromatin remodeling and DNA replication. The G2/M phase was enriched in proteins involved in protein synthesis and microtubule dynamics, essential for mitosis. Notably, metabolic enzymes represented approximately 6% of the proteins identified in the purified nuclear fraction, consistent with previously published proteomic analyses of T. cruzi nuclear-enriched fractions. Hexokinase, citrate synthase, and pyruvate kinase activities were detected in extracts from this fraction, while immunofluorescence supported the association of hexokinase and citrate synthase with the nuclear region. These results suggest that metabolic enzymes and/or their metabolites may modulate nuclear processes in T. cruzi, potentially influencing the epigenome and gene expression regulation, as proposed in other eukaryotic models. Altogether, our findings reveal dynamic remodeling of the nuclear proteome during the T. cruzi cell cycle and point to a previously underappreciated role for metabolic enzymes likely regulating nuclear functions in a cell cycle-dependent manner.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0013299"},"PeriodicalIF":3.6,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13645223/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149076423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pre-extensively and extensively drug-resistant tuberculosis prevalence among multidrug-resistant tuberculosis cases in Ethiopia: A systematic review and meta-analysis.","authors":"Muluneh Assefa, Mitkie Tigabie, Abebaw Setegn, Yenesew Mihret Wondmagegn, Azanaw Amare, Getu Girmay, Sirak Biset, Wesam Taher Almagharbeh","doi":"10.1371/journal.pntd.0014744","DOIUrl":"10.1371/journal.pntd.0014744","url":null,"abstract":"<p><strong>Background: </strong>The burden of pre-extensively drug-resistant tuberculosis (Pre-XDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) cases in Ethiopia has become one of the most challenging problems for monitoring the treatment of patients with multidrug-resistant tuberculosis (MDR-TB). This study aimed to provide a valuable evidence synthesis on the pooled prevalence of Pre-XDR and XDR-TB among MDR-TB patients in Ethiopia.</p><p><strong>Methodology: </strong>This study was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Extracted data from relevant articles were analyzed using STATA version 17.0. The pooled estimate of the effect size was computed using a restricted maximum likelihood random-effects model, considering a 95% confidence interval, and a forest plot was used to visualize the proportion of Pre-XDR and XDR-TB. Because the Pre-XDR and XDR-TB cases are rare, the Freeman-Tukey double arcsine transformation was applied to estimate the pooled proportions. The I2 statistic and Galbraith plot confirmed heterogeneity. A univariate meta-regression, sensitivity, and subgroup analyses were conducted to identify the source of heterogeneity. Egger's test and funnel plot were used to check publication bias.</p><p><strong>Results: </strong>Eight studies comprising 1,518 MDR-TB patients, with 656 new and 862 previously treated cases, were included. Among those, 52 Pre-XDR and 13 XDR-TB cases were identified. According to the Freeman-Tukey transformation, the overall pooled prevalence of Pre-XDR-TB was 3% (95% CI: 2%-5%, I2 = 43.69%) and XDR-TB was 1% (95% CI: 0%-4%, I2 = 54.07%). In the subgroup analysis, the prevalence of Pre-XDR-TB across studies from multiple regions of Ethiopia was 7% (95% CI: 3%-13%), and in Addis Ababa, it was 3% (95% CI: 2%-4%). Single studies in the Amhara, Tigray, and Oromia regions reported Pre-XDR-TB prevalence of 6%, 5%, and 3%, respectively. The pooled prevalence of XDR-TB in Addis Ababa was 1% (95% CI: 0%-2%). A single study conducted in Oromia reported a relatively higher prevalence of XDR-TB; 10% (95% CI: 3%-26%). Regarding the study year, the XDR-TB prevalence was 3% (95% CI: 0%-11%) in 2005-2018 and 1% (95% CI: 0%-2%) in 2019-2024. A statistically significant publication bias was observed in XDR-TB prevalence (p = 0.024).</p><p><strong>Conclusion: </strong>There is a significant prevalence of Pre-XDR and XDR-TB cases among MDR-TB patients in Ethiopia. It highlights the need for strengthened surveillance in all regions and expanded drug susceptibility testing of local strains for better management of MDR-TB patients.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014744"},"PeriodicalIF":3.6,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13638038/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149076400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Implementation of syndromic surveillance systems in two rural villages in Senegal.","authors":"","doi":"10.1371/journal.pntd.0014748","DOIUrl":"10.1371/journal.pntd.0014748","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1371/journal.pntd.0005212.].</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014748"},"PeriodicalIF":3.6,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13618899/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149076433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stage-specific remodeling of the pulmonary microenvironment during Paragonimus proliferus infection in a rat model.","authors":"Danhong Cheng, Lei Zhang, Le Sun, Min Yin, Yu Wang, Shuwen Yang, Xing Yan, Xiaoyan Zhu, Cuiying Li, Weixun Chunyu","doi":"10.1371/journal.pntd.0014760","DOIUrl":"10.1371/journal.pntd.0014760","url":null,"abstract":"<p><p>Paragonimiasis, a food-borne zoonosis caused by Paragonimus spp., can cause severe pulmonary inflammation and fibrosis. However, relationships among the host, parasite and lung microbiome at defined infection stages remain poorly understood. We compared independent groups of rats sampled at 14, 28 and 42 days post-infection (dpi), together with a separate group sampled after triclabendazole (TCBZ) treatment. Histopathological and molecular analyses were combined with multi-region (5R) 16S rRNA gene sequencing and fluorescence in situ hybridization (FISH). Groups sampled at later post-infection time points showed greater pulmonary inflammation and collagen deposition, with the highest values in Pp-42d; both outcomes were lower in Pp-TCBZ than in Pp-42d. Th1-, eosinophil-, Th2- and Treg- associated markers differed among experimental groups. TLR4/NF-κB -related protein expression and pulmonary microbial profiles also differed among groups including enrichment of taxa such as Lactobacillus in infected animals. FISH detected bacterial signals within inflammatory lesions and Lactobacillus-associated signals spatially associated with parasite eggs. These repeated cross-sectional findings identify stage-associated differences in pulmonary pathology, immune markers and microbiota during P. proliferus infection, together with lower pathological measures in the Pp-TCBZ group than in the untreated Pp-42d group. The egg-associated bacterial signals warrant further investigation but do not establish a functional host-parasite-microbiome mechanism.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014760"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13626465/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Barriers in operating proactive source reduction informed by an early warning system for dengue prevention: A qualitative study from end-user perspectives in Vietnam.","authors":"Kien Quoc Do, Ha Thu Nguyen, Thi Thanh Thao Nguyen, Cong Tuan Pham, Nguyen T T K, Dang Khanh Linh Vien, Dung Phung","doi":"10.1371/journal.pntd.0014754","DOIUrl":"10.1371/journal.pntd.0014754","url":null,"abstract":"<p><strong>Background: </strong>Dengue is a major vector-borne disease, with rising incidence driven by climate change, rapid urbanization, and increased international travel. Although early warning systems (EWSs) have shown promising predictive performance in endemic settings, limited evidence on implementation barriers from local end users hinders their integration into routine proactive vector control.</p><p><strong>Methods: </strong>A qualitative study with in-depth interviews was conducted with local end users, including local authorities, health practitioners, and health volunteers. A deductive approach utilizing a predefined conceptual framework was employed to code the verbatim transcripts. Inductive coding was subsequently incorporated to capture emerging concepts that were not represented in the initial framework. Thematic analysis was performed to analyse the collected data. To enhance the methodological rigor, interviews and coding were performed by multiple and diverse researchers from various academic and practice institutions, and the transcripts were validated through member checking with selected participants.</p><p><strong>Results: </strong>The lack of supportive policies, limited engagement from relevant stakeholders due to low familiarity with EWS concepts, and operational challenges were identified as critical barriers to implementing EWS-informed proactive source reduction. The perception that dengue risks were only during the rainy season, together with socio-economic constraints, water scarcity exacerbated by extreme weather events, and the application of inappropriate source reduction measures, further constrained household participation. The multi-level barriers collectively disrupted the translation of EWS alerts into proactive dengue prevention.</p><p><strong>Conclusions: </strong>Early warning information alone is insufficient to translate into effective and sustained proactive vector control for dengue prevention. Strengthening enabling policies, local stakeholder and community engagement, and awareness of EWS-informed risk management across multiple levels are essential to translate EWS alerts into timely and sustained vector control actions in dengue-endemic settings similar to Vietnam. Further studies across diverse settings are needed to assess the broader applicability of EWS-informed approaches.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014754"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13649133/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effectiveness of community-based delivery of mass dog vaccination to prevent rabies: A cluster randomized controlled trial.","authors":"Felix Lankester, Ahmed Lugelo, Joel Changalucha, Danni Anderson, Christian Tetteh Duamor, Anna Czupryna, Kennedy Lushasi, Elaine Ferguson, Emmanuel S Swai, Maganga Sambo, Jonathan Yoder, Sarah Cleaveland, Sally Wyke, Benezeth Lutege Malinda, Paul C D Johnson, Katie Hampson","doi":"10.1371/journal.pntd.0014704","DOIUrl":"10.1371/journal.pntd.0014704","url":null,"abstract":"<p><strong>Background: </strong>Approximately 60,000 people die from rabies annually, mostly in Africa and Asia. Mass dog vaccination is critical for elimination. Herd immunity has been calculated to be attained when >40% (critical threshold) of dogs are vaccinated. A common delivery method implemented in Tanzania and other countries typically involves non-local teams travelling to villages implementing dog vaccination annually through static-point clinics using cold chain stored vaccines (Team-based delivery). Determination that a canine rabies vaccine is thermotolerant enables novel Community-based dog vaccination strategies whereby community vaccinators use vaccines stored locally throughout the year (Community-based delivery).</p><p><strong>Methods: </strong>Two dog vaccination strategies were compared using a three-year cluster randomized controlled trial in Tanzania (Clinical Trials Registration Number ISRCTN14813279). The strategies were designed to represent implementation using resources typical of government-led initiatives. Wards (n = 112) were randomly assigned to either Team-based delivery or Community-based delivery. Coverage was estimated using household surveys implemented biannually.</p><p><strong>Findings: </strong>Mean coverage achieved by Community-based delivery (55%; 95% Confidence Interval (CI): 45-65%) was higher than Team-based delivery (37%; 95% CI: 28-46%). Moreover, because coverage was lower and declined over each year only in Team-based delivery, the probability of coverage being below the critical threshold was higher for Team-based delivery (60%) compared to Community-based delivery (18%). To ensure coverage does not dip below the critical threshold by the year end, coverage at the start of the year in Team-based delivery needed to be higher (61%) than in Community-based (42%).</p><p><strong>Interpretation: </strong>Community-based delivery achieved higher and more consistent vaccination coverage across a range of settings typical of many sub-Saharan African countries. Generalisation to different sociocultural/ agroecological settings requires further evidence. Although direct evidence on impact on rabies elimination was not measured, this approach could play a role in national elimination strategies, developed for the global 'Zero by 30' strategy to end dog-mediated rabies deaths by 2030, and in response to investment by Gavi, the vaccine alliance.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014704"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13641700/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early warning of serotype replacement, spatial risk redistribution, and shifting clinical phenotypes: Insights from the 2023-2025 dengue outbreak in Meta, Colombia.","authors":"Liliana Marcela Montilla Rodríguez, Cesar García-Balaguera, Sandra Campo Palacio, David Bonilla Montes, Yuri Daniela Cadena Díaz, Paula Alejandra Mosquera Tocora, Norma Cristina Pavas Escobar, Camilo Andrade Carreño, Elsa Victoria Andrade Miranda, Marina Sthella Gonzalez Robayo","doi":"10.1371/journal.pntd.0014733","DOIUrl":"10.1371/journal.pntd.0014733","url":null,"abstract":"<p><strong>Background: </strong>Dengue remains a major public health challenge in hyperendemic regions, where shifts in serotype circulation, spatial transmission patterns, and clinical presentation can significantly influence outbreak dynamics. Colombia has experienced intense dengue activity in recent years; however, integrated analyses combining virological, spatial, and clinical data remain limited.</p><p><strong>Methods: </strong>A retrospective longitudinal observational study was conducted using routinely collected dengue surveillance data from the Department of Meta, Colombia, covering the period from January 2023 to September 2025. Confirmed dengue cases with viral isolation and serotype identification were included. Temporal trends, serotype replacement dynamics, municipal-level spatial redistribution patterns, and clinical characteristics were analyzed using laboratory-confirmed dengue cases. Multivariable logistic regression models were used to identify factors associated with dengue positivity overall and by serotype, while interannual variation was assessed to evaluate changes in transmission intensity and the spatial redistribution of dengue transmission risk.</p><p><strong>Results: </strong>Marked serotype replacement was observed during the study period. DENV-2 predominated during the 2023 peak, followed by a decline and a sustained increase in DENV-3 and DENV-4 by 2025, while DENV-1 disappeared from recent records. Spatial analysis showed declining diagnostic positivity in historically hyperendemic municipalities and significant interannual increases in intermediate municipalities, indicating an epidemiological rebound outside major urban centers. Classical dengue symptoms, including headache, myalgia, and rash, were inversely associated with virological confirmation, whereas nonspecific neurological manifestations, such as somnolence and irritability, emerged as independent predictors. Severe thrombocytopenia remained strongly associated with infection, particularly in DENV-2 cases.</p><p><strong>Conclusions: </strong>The dengue outbreak in Meta (2023-2025) was characterized by dynamic serotype replacement, spatial redistribution of transmission risk, and changing clinical presentations. The rapid emergence of DENV-4 in 2025 serves as an early warning of serotype replacement and underscores the need to update clinical screening protocols to include nonspecific neurological manifestations. Strengthening decentralized laboratory networks and closing surveillance gaps in low-endemicity areas are strategic priorities to improve early detection and outbreak control in hyperendemic regions.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014733"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13614597/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Flubendazole reduces emerging Dracunculus medinensis in dogs in Chad: Results from a two-year study.","authors":"Amy C Dupper, Charles D Mackenzie, Timothy G Geary, Deborah L Elder, Wided Najahi-Missaoui, Christopher A Cleveland, Ellen K Haynes, Michael J Yabsley, Guilherme Gomes Verocai, Macon Overcast, John A Bryan Ii, Fernando Torres-Velez, Métinou Koumétio Sidouin, Bongo Nare Richard Ngandolo, Philip Tchindebet Oaukou, Adam J Weiss, Rebecca Garabed","doi":"10.1371/journal.pntd.0014763","DOIUrl":"10.1371/journal.pntd.0014763","url":null,"abstract":"<p><p>Since being targeted for eradication in the 1980s, global human cases of dracunculiasis (Guinea worm disease; GWD) have been reduced by over 99.9% through interventions aimed at interrupting the life cycle of the causative parasite, Dracunculus medinensis. However, the burden of GWD in dogs is higher than in humans and other monitored mammalian host species, threatening eradication of GWD. Expanding on a previous clinical trial of flubendazole (FLBZ) in dogs in Chad, we conducted a cluster-randomized controlled trial in which half of enrolled villages (N = 29) were assigned to receive FLBZ and the other half (N = 27) to receive no treatment. An annual difference-in-differences analysis revealed that treated villages had 59.3% fewer predicted emerging D. medinensis adult females (95% CI: 53.8% fewer to 69.2% fewer) per village per year during the second year of the study compared to control villages. Analyses examining effects of FLBZ monthly, at the individual dog level, and through histopathology of D. medinensis female reproductive morphology, all support the conclusion that FLBZ inhibits embryogenesis and larval development within the uterus of gravid female worms. Together these data are consistent with a hypothesis that although D. medinensis emerged from individually treated dogs, the L1s released from dogs treated with FLBZ exhibited reduced viability, thus reducing the risk of infection in the next mammalian host. This trial builds the body of knowledge about FLBZ's impacts on D. medinensis in the canine host and provides compelling evidence on the effectiveness of FLBZ in reducing the number of emerging D. medinensis in dogs, which can contribute to the eradication of this parasite. However, additional questions remain regarding the formulation, which need to be addressed in future studies before being deployed at scale.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014763"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13630316/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integration of vector and human behavior in residual malaria in rural communities in the Peruvian Amazon.","authors":"Carlos Acosta, Diana Cubas-Montecino, Joaquin Gomez, Marlon Saavedra, Mitchel Guzman-Guzman, Sara A Bickersmith, Jan E Conn, Dionicia Gamboa, Joseph M Vinetz","doi":"10.1371/journal.pntd.0014261","DOIUrl":"10.1371/journal.pntd.0014261","url":null,"abstract":"<p><strong>Background: </strong>Malaria persists as a leading cause of morbidity in the Amazon Basin, where the Loreto region of Peru contributes over 90% of the national burden. Despite widespread deployment of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) under Peru's Malaria Elimination Plan 2022-2030, residual transmission continues in rural riverine communities. This transmission is driven by the interplay between vector ecology and human behavior. This study quantified residual human exposure to Nyssorhynchus darlingi by integrating hourly entomological indices with real-time human behavioral data across two seasons in three communities in Mazán, Loreto.</p><p><strong>Methodology/principal findings: </strong>Entomological (HLC) and epidemiological (HBS) data were collected during the wet and dry seasons of 2022. Plasmodium infections were detected in 619 participants by microscopy and qPCR. Molecular surveillance revealed a 13% infection prevalence over seven times higher than routine microscopy (2%) indicating a substantial submicroscopic reservoir. Infection burden was heterogeneous, with Gamitanacocha showing the highest prevalence (33%). LLIN usage was high (>97%), yet nets prevented only 63.3%-80.3% of potential mosquito bites. Residual exposure occurred mainly indoors during early evening (18:00-21:00) while individuals were awake and unprotected, particularly in the dry season when domestic routines coincided with peak vector biting. Outdoor exposure increased during the wet season due to evening recreational activities. Multivariate analysis identified age as the only significant predictor of infection (p < 0.05), with individuals over five years showing consistently higher odds of infection than children under five (OR range: 3.62-5.46).</p><p><strong>Conclusions/significance: </strong>A critical protection gap exists during early evening and dawn, when human activity overlaps with peak vectors biting outside the protection of bed nets. The high submicroscopic burden and persistent indoor exposure while awake demonstrate that current interventions are insufficient for elimination. These findings highlight that Loreto's elimination strategies must complement intradomiciliary tools with peri-domestic and behavioral interventions-such as full house screening and spatial repellents-specifically targeting the exposure windows identified here.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014261"},"PeriodicalIF":3.6,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13638056/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149065623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and evaluation of a multiplex PCR for simultaneous detection of Mycobacterium tuberculosis complex, Mycobacterium avium complex, and Burkholderia pseudomallei in clinical specimens.","authors":"Hasyanee Binmaeil, Thanakon Bunsong, Janjira Thaipadungpanit, Wipawee Saenwongsa, Montira Ngoenpramual, Sutthinan Chawsorngkorn, Soawarat Deekae, Sunee Chayangsu, Wang Nguitragool, Narisara Chantratita","doi":"10.1371/journal.pntd.0014730","DOIUrl":"10.1371/journal.pntd.0014730","url":null,"abstract":"<p><p>Effective control of tuberculosis, nontuberculous mycobacterial (NTM) disease, and melioidosis depends on rapid and accurate laboratory diagnosis. Mycobacterium tuberculosis complex (MTBC) remains a health priority, whereas NTM infections, particularly Mycobacterium avium complex (MAC), are increasingly recognized and may be misdiagnosed as tuberculosis. Melioidosis, caused by Burkholderia pseudomallei, is underdiagnosed in endemic regions. These infections have overlapping clinical presentations, and conventional diagnostics methods may not accurately differentiate among these diseases. We developed and evaluated a quadruplex real-time PCR assay (TAM-mPCR) for simultaneous detection of MTBC, MAC, and B. pseudomallei, targeting IS6110, rrs, and BPSS1187. A total of 607 pulmonary and extrapulmonary specimens from patients with suspected tuberculosis were collected at two sites in Thailand between 2024 and 2025. The limits of detection were 2 pg/reaction for MTBC, 0.1 ng/reaction for MAC, and 50 fg/reaction for B. pseudomallei, with 100% analytical specificity. For MTBC detection, TAM-mPCR demonstrated 100% sensitivity (330/330) and 99% (189/191) specificity. All three culture-confirmed MAC specimens were detected. For B. pseudomallei, the assay demonstrated 73.2% sensitivity (30/41) and 100% specificity (79/79). For MTBC, TAM-mPCR showed high concordance with NAAT and culture results, but differed significantly from AFB microscopy (P < 0.001). The assay also detected additional MTBC, MAC, and B. pseudomallei infections that were missed by routine diagnostics. TAM-mPCR enables rapid simultaneous pathogen detection and differentiation and may improve case detection and patient management in setting where tuberculosis and melioidosis co-endemic.</p>","PeriodicalId":49000,"journal":{"name":"PLoS Neglected Tropical Diseases","volume":"20 9","pages":"e0014730"},"PeriodicalIF":3.6,"publicationDate":"2026-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13606991/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149041475","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}