Despite the widespread distribution of Trypanosoma evansi in camels and horses in Iran, only a few case reports of canine infection are available. We investigated the prevalence of trypanosomosis in 423 apparently healthy sheltered stray and owned dogs from five provinces representing different climatic regions of Iran (Hamedan, Kermanshah, Mazandaran, Yazd, and Khuzestan). Blood samples were examined microscopically and analyzed by PCR targeting the RoTat 1.2 variant surface glycoprotein (VSG) gene of T. evansi. RoTat-PCR detected T. evansi DNA in 97 dogs (22.9%), with provincial prevalences ranging from 7.5% to 39.0%; representative amplicons were confirmed by sequencing and phylogenetic analysis. No trypomastigotes were detected by microscopic examination. This study provides the first molecular evidence of T. evansi DNA in dogs from Iran and demonstrates widespread molecular detection of T. evansi among clinically healthy dogs. These findings suggest that exposure of dogs to T. evansi in endemic regions may be more common than previously recognized. However, the epidemiological significance of PCR-positive dogs remains uncertain. Future studies integrating parasite isolation, genomic characterization, longitudinal monitoring, and transmission investigations will be essential to determine the biological significance of these molecular findings and to clarify the epidemiological significance of asymptomatic PCR-positive dogs.

Cryptosporidiosis causes a significant impact on immunocompromised patients, but its epidemiology among Egyptian cancer patients is poorly studied. This study was conducted to evaluate the prevalence, species distribution, and Cryptosporidium DNA copy number among Egyptian cancer patients versus immunocompetent controls. This case-control study included 600 cancer patients and 400 immunocompetent controls. In addition to modified acid-fast staining, molecular analysis was performed using eight new, in silico and in vitro validated species-specific primer sets targeting Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium meleagridis, Cryptosporidium muris, Cryptosporidium canis, Cryptosporidium felis, Cryptosporidium ubiquitum, and Cryptosporidium chipmunk, using a qPCR-based standard curve. Cryptosporidium infection was found in 37.0% of cancer patients compared to 11.8% of controls (p< 0.0001). C. parvum was more prevalent among the cancer patients (62.5%) than among controls (46.8%, p= 0.009), whereas C. muris was more prevalent among controls (10.6% vs. 1.5%, p= 0.008). qPCR analysis indicated significant differences in Cryptosporidium DNA copy number (copies/µL) of C. parvum and C. hominis (p< 0.05). Species distribution varied significantly with cancer type (p< 0.001); C. parvum was present only in acute lymphocytic leukemia (ALL), gastric adenocarcinoma, colorectal, and liver cancers, whereas C. hominis was found in acute myeloid leukemia (AML). C. meleagridis and C. muris were identified in patients with acute lymphocytic leukemia and colon cancer. Our results showed a significantly higher prevalence of the parasite in cancer patients, with a notable difference in the distribution of species depending on the type of malignancy and clinical relevance for targeted screening.

Kudoa bora (Fujita, 1930) is a myxosporean parasite primarily infecting the skeletal muscle of mugilid fishes. In this study, we report for the first time that K. bora can infect the periocular tissues of Osteomugil perusii (Valenciennes, 1836). A comparative transcriptomic approach was employed to preliminarily characterize the molecular responses of the host's periocular tissues to the parasitic infection. A total of 2,954 differentially expressed genes (DEGs) were identified between the infected and uninfected groups, of which 1,305 were upregulated and 1,649 were downregulated. Enrichment analysis of the DEGs revealed that metabolic pathways such as ribosome biogenesis, oxidative phosphorylation, and DNA repair were significantly upregulated, suggesting that the host may activate protein translation and energy compensation mechanisms under stress induced by K. bora infection. In contrast, the cytokine-cytokine receptor interaction and TGF-β signaling pathways were markedly downregulated, suggesting that periocular infection may be associated with a certain degree of local immune suppression. Widespread downregulation of genes involved in eye morphogenesis, lens structural proteins, and visual signal transduction was observed, potentially compromising visual function. Collectively, periocular infection by K. bora may compromise host visual function through the combined suppression of immune pathways and vision-related gene expression. This study extends the known parasitic site of K. bora and provides preliminary molecular evidence for pathogenic mechanisms of myxosporean infection in fish periocular tissues.

Neglected tropical parasites including soil-transmitted helminths, food-borne helminths and schistosomes remain a public health concern across much of the world including Southeast Asia. Abiotic and biotic environmental drivers not only directly impact helminth transmission but are also essential parameters for stable intermediate host ecologies. These environmental risk factors thus must be considered for disease control. To better understand the role of the environment, ongoing and new areas of research and the expansion of monitoring techniques such as environmental DNA are on the rise. This review will highlight the importance of environmental factors and influences in the survival and transmission of helminths directly and the distribution of their intermediate hosts. Focusing on research based in Southeast Asia due to the region's ideal climate for parasite disease transmission. The diversity of tropical and subtropical climates and environmental conditions, with warmer temperatures and higher humidity, promotes helminth lifecycles, including the requirements of intermediate hosts. We discuss how research into helminth ecology provides an improved understanding of how to monitor and control regionally endemic helminths. Moreover, the integration of environmental drivers with eDNA approaches offers a promising new framework for linking helminth ecology and environmental surveillance.

Insecticide resistance profiles and underlying mechanisms are essential for developing effective and sustainable vector control strategies. This study aimed to investigate the insecticide susceptibility profiles and kdr genotypes in field populations of Aedes aegypti and Ae. albopictus. Bioassays were conducted on six Ae. albopictus and two Ae. aegypti populations towards deltamethrin, lambda-cyhalothrin, pirimiphos-methyl, and bendiocarb. Piperonyl butoxide (PBO) synergism and kdr genotyping of the Vssc gene were conducted. Against pyrethroids, Ae. aegypti and four out of six Ae. albopictus populations exhibited resistance. PBO restored full susceptibility across Ae. albopictus populations but not against Ae. aegypti, suggesting metabolic resistance involving cytochrome P450 monooxygenese. All tested mosquitoes were susceptible to pirimiphos-methyl. F1534C and V1016G kdr mutations were detected in both populations of Ae. aegypti. For the F1534C locus, the resistant allele Cys1534 was predominant but population specific. At codon 1016, Gly1016 was present but less frequent than Val1016, with resistant homozygotes population specific. These findings highlight heterogeneous resistance profiles across provinces and species, underscoring the need for vector and location-specific vector control strategies. PBO synergist addition remains viable option for managing Ae. albopictus resistance but pyrethroid alternatives such as pirimiphos-methyl and bendiocarb offer an interspecies option for controlling Aedes vectors.

New World phlebotomine sand flies include medically important vectors and taxonomically complex species groups. Public COI barcode libraries are increasingly used for species identification, biodiversity surveys and vector surveillance, yet their systematic reliability has not been evaluated at continental scale. Here we integrate GenBank and BOLD COI records for New World Phlebotominae to assess taxonomic and geographic coverage, metadata quality, barcode-gap performance, BIN consistency and phylogenetic signal in priority species complexes. We curated 6,200 COI or COI-like records, including 5,562 records assigned to binomial species. The analysis-ready dataset comprised 5,133 filtered records, 3,671 unique haplotypes and 224 species distributed among 20 operational genus-level database labels, all mapped to the Galati et al. (2025) world-checklist hierarchy for supraspecific interpretation. Metadata completeness was uneven: 57.03% of records had country information, 54.40% had coordinates, 64.02% had a BOLD BIN, 82.65% had voucher information and 69.55% had a reference title. Among 203 species evaluable for barcode-gap analysis, 123 showed a barcode gap and 80 showed absent or overlapping gaps. Integrating barcode gap, BINs, geography and focal maximum-likelihood trees classified 103 species as supported by COI for species assignment, 38 as barcode identification risks, 34 as candidates for cryptic diversity or structured populations, 19 as probable species complexes or COI-limited cases, 9 as high-priority species-boundary problems and 21 as insufficiently sampled. Focal IQ-TREE analyses of six critical groups showed strict non-monophyly in 29 of 35 evaluated species. Nearest-neighbour analysis recovered 137 priority species pairs for systematic review; 59 were placed in the same Galati subgenus and 32 in the same species series or group. COI is therefore useful for many New World sand fly species, but its strongest current value is as an audit tool for reference libraries and as a source of evolutionary hypotheses about species groups, sibling species and geographically structured lineages. We provide a reproducible framework for converting public barcode data into a vector-identification and systematic-triage agenda for Phlebotominae.

Malaria elimination requires distinguishing continued reporting from return after zero-reporting periods. In this longitudinal ecological study, we compared monthly persistence and reappearance of reported Plasmodium falciparum and Plasmodium vivax malaria in the Peruvian Amazon from 2009 to 2024. Reported surveillance episodes, which were not assumed to represent unique individuals, were aggregated by month, species, and 350 harmonized geographic units while retaining zero counts. A dynamic hurdle model combined a Bernoulli occurrence component, a zero-truncated negative binomial positive-count component, species-specific spatial effects, and geospatial and temporal covariates. For temporal validation, models were fitted on 2009-2022 and evaluated in 2023-2024. Among 585,510 reported surveillance episodes, 81.8% were attributable to P. vivax. In the 76-unit eligible common-support universe, adjusted persistence was 0.790 for P. falciparum and 0.848 for P. vivax; the difference was 0.053 (approximate 95% uncertainty interval [UI], -0.088 to 0.299). Under the corrected reappearance risk set requiring a strictly earlier positive report, adjusted reappearance was 0.071 and 0.146, respectively; the difference was 0.075 (approximate 95% UI, 0.030 to 0.128). Predicted-to-observed ratios were 0.821 for P. falciparum and 0.900 for P. vivax; both met the prespecified validation criteria, although underprediction was greater for P. falciparum. Larger population, lower elevation, and greater forest cover were associated with reported occurrence. The persistence contrast remained uncertain; corrected grouped reappearance was higher for P. vivax, although duration-specific contrasts did not uniformly exclude zero. Separating surveillance-defined continuity from return of reporting may support subnational monitoring after prospective validation; climate associations remained exploratory because of inadequate calibration and instability.

Dengue virus (DENV) poses a growing global public health threat due to increasingly frequent and severe outbreaks. Moreover, epidemiological surveillance systems have limitations in the timely detection of outbreaks, highlighting the need for complementary tools such as Wastewater-Based Epidemiology (WBE). This study aimed to generate further information on the persistence and recovery of DENV RNA in urban wastewater to support its monitoring through WBE. Two experiments were conducted. In Experiment 1, the persistence of DENV RNA in urban wastewater incubated at 4, 25, and 35 °C for 21 days was evaluated. A negative binomial mixed-effects regression model estimated temperature-specific decay coefficients of -0.67 (95 % confidence interval (CI): -0.74--0.59), -1.25 (95 % CI: -1.32--1.17), and -1.31 (95 % CI: -1.37--1.23) at 4, 25, and 35 °C, respectively. The corresponding times required for a 90 % reduction in genomic copies concentration were 30.1 (95 % CI: 20.7-48.1), 5.3 (95 % CI: 4.7-6.1), and 4.8 (95 % CI: 4.3-5.4) days. In Experiment 2, the performance of four recovery methods was compared. Ultrafiltration showed the highest recovery efficacy (61 ± 8 %), followed by direct analysis (42 ± 10 %) and polyethylene glycol precipitation (7 ± 4 %). Skimmed milk flocculation yielded results below the limit of detection. In conclusion, DENV RNA persisted in urban wastewater for several days at environmentally relevant temperatures, and ultrafiltration achieved the highest recovery efficacy among the methods evaluated, supporting the application of WBE as a community surveillance tool for DENV.

Dengue remains a major public health concern in Indonesia, and despite recent progress in vaccine development, vector control remains essential to limit dengue outbreaks and occurrence of clinical cases. Among emerging approaches, the radiation-based sterile insect technique (SIT) has gained attention as an environmentally friendly vector control strategy. This review synthesizes two decades (2004-2024) of SIT research and development against Aedes aegypti in Indonesia, tracing the programme's evolution from unstructured pilot releases to a structured phased conditional approach (PCA) framework that culminated in a recent small-scale field trial in Bandung city. Taking the field trial as the central case study, we identify a core challenge: while SIT can significantly reduce vector populations, suppression remains incomplete and impermanent without optimized integration, as evidenced by post-release population rebound. Further synthesized challenges include navigating the impact of geographical genetic diversity on mating competitiveness, ensuring long-term programme sustainability through stakeholder commitment, and cost-effectiveness concerns for resource-limited settings. To guide the transition toward the pre-operational Phase 3, we propose a dedicated, prioritized research agenda including cluster-randomized controlled trials with entomological and epidemiological endpoints, factorial designs to isolate intervention effects, and development of genetic sexing strains and tailored release protocols, to improve cost-efficiency and prevent population rebound. This synthesis provides a programmatic framework for interpreting Indonesia's SIT development and may inform other dengue-endemic countries considering SIT integration into vector-control programmes.

Cutaneous Leishmaniasis (CL) is a vector-borne parasitic disease closely linked to environmental conditions. This study aimed to model and map the spatial occurrence potential of CL across multiple study years in Golestan Province, Iran, using machine learning, ensemble learning approaches, and remote sensing data. A geospatial database was developed from CL case records aggregated at the level of cities and rural settlements during 2011-2013 and nine environmental variables, including precipitation, temperature, evapotranspiration, wind speed, drought conditions, vegetation cover, population density, elevation, and slope. Three machine learning algorithms-Random Forest (RF), Support Vector Regression (SVR), and Artificial Neural Network (ANN)-were applied. The outputs of the machine learning models were combined using majority voting and weighted voting techniques. Model performance was evaluated based on the Area Under the ROC Curve (AUC). Among individual models, ANN showed the highest performance (AUC = 0.843), while SVR had the lowest (AUC = 0.748). Ensemble approaches improved overall discriminatory performance, with weighted voting achieving the best overall performance (AUC = 0.911). Spatial results indicated that the northern and central areas had the highest CL occurrence potential, whereas the southern and southeastern regions had the lowest occurrence potential. These findings highlight the potential of integrating remote sensing data, machine learning algorithms, and ensemble methods for CL occurrence mapping.

In low- or middle-income countries, neglected tropical diseases like taeniasis/cysticercosis from Taenia solium have serious health, social and economic consequences. Despite urgent calls from WHO and governments for intensified control of T. solium, this remains challenging in practice. Intervention failures point to gaps in understanding about behaviours relating to T. solium risk and the critical need for insight into these behaviours. In this study we explored known behavioural risks associated with T. solium in northern Lao PDR. A qualitative assessment was conducted in 28 villages using group discussions, interviews and observation involving 438 people (female:male ratio of 35:65). The study identified a range of contextual and socio-cultural factors influencing risky behaviours, including previously unreported aspects. Raw pork is consumed by people from all four main ethnolinguistic groups, with raw pork dishes commonly available at traditional events, but less commonly part of ceremonial practice. Consumption of raw wild pork was frequently reported across the study villages and often preferred because of its taste. Open defecation is most common in places where toilet coverage is low, but the protective factor of toilets is undermined by time spent out of the village and seasonal water shortages. Free-roaming pigs are common seasonally, but less common when sanctions are in place and enforced. However, there are circumstances that result in a relaxing of rules including feed shortages, inadequate pens and traditional beliefs. Understanding these behavioural risks is essential for developing effective strategies to change behaviour and control/eliminate T. solium.

The family Trypanosomatidae includes parasitic genera of medical importance, such as Leishmania and Trypanosoma, which cause leishmaniasis and Chagas disease, respectively. Although these parasites have complex life cycles involving insect vectors and mammalian hosts, the role of bats in these cycles remains poorly understood. This study aimed to identify Leishmania and Trypanosoma species associated with bats in an endemic area in southeastern Veracruz, Mexico. Bats were captured with mist nets at three tropical sites, and species were identified morphologically. Molecular detection was performed by PCR amplification of the 18S rDNA and ITS-1 regions. Overall, 43 bats belonging to eight species and three families were examined. Trypanosomatid DNA was detected in four individuals (9.3%; 4/43). Leishmania mexicana was identified in two specimens of Pteronotus fulvus (4.6%; 2/43). Leishmania infantum was detected for the first time in Pteronotus mesoamericanus (2.3%; 1/43). Trypanosoma cruzi was identified in a single specimen of Choeroniscus godmani (2.3%; 1/43). These findings expand the known host range of trypanosomatids in Mexico and suggest that bats may play an underrecognized role in maintaining these parasites in endemic tropical regions.

Triatoma infestans, the primary domiciliary vector of Chagas disease in Chile, has a broad geographic distribution and is characterized by high synanthropy and strong colonization capacity. Landscape characteristics are key factors in understanding population distribution and the processes that shape morphological variation. This study aimed to characterize the structures associated with dispersal and feeding using geometric morphometrics in domiciliary adults of T. infestans (n = 353 total specimens analyzed) along a latitudinal gradient (∼1255 km). Furthermore, the specimens' localities were classified according to the Köppen-Geiger climatic regionalization, resulting in four ecoregions. The relationship between morphology and climatic variables was evaluated using bioclimatic CHELSA data (1981-2010). Cold Desert (northern distribution) individuals exhibited more compact shapes than those of the Semi-arid and Mediterranean (southern distribution) ecoregions. Pronotum centroid size remained stable across all ecoregions (p = 0.095) and between sexes (p = 0.443). The head centroid size showed sexual dimorphism in some ecoregions, with females larger than males. Females also exhibited morphological stability, which could be associated with feeding prioritization and reproductive investment rather than dispersal. In contrast, lateral head centroid size in males varied with latitude, being smaller in the Cold Desert ecoregion than the Mediterranean ecoregion (p < 0.01). Macroclimatic factors did not explain shape variation (p > 0.05 for all traits). The results suggest a modular response between the pronotum and the head, potentially driven by the microenvironmental conditions within domestic habitats. Such patterns highlight the importance of trait-specific morphological variations, which may have implications for dispersal and feeding, and could be integrated into surveillance and vector control strategies.

Leishmaniasis has been endemic in Syria for hundreds of years. Cutaneous Leishmaniasis (CL) has become an urgent public health challenge in the conflict-affected Northwestern Syria (NWS), where there has been significant community displacement, healthcare deterioration, and environmental degradation. We investigated the socio-economic, health awareness, psychosocial, and environmental awareness of community members afflicted with CL in the Aleppo region (NWS). A questionnaire was completed in June 2024 by 320 randomly selected members of the community with active or past CL in the districts of Atareb, Daret Azza, and Afrin, including 130 internally displaced people (IDP) and 38 individuals living in camps. Focus group discussions were held with public health center staff in the same districts. Data was analyzed using Bayesian log-linear modeling and one-way ANOVA. Many respondents have good understanding about the disease, with 88.44% of respondents receiving information about CL. Despite misconceptions regarding CL transmission 28% of respondents accurately identified bites of sand flies as the main reason for infection. Sleeping indoors and conducting sand fly control were associated with a lower likelihood of past leishmaniasis. 43% of participants reported that they had to take leave from work, leading to severe financial strains exacerbated by CL treatment costs averaging $135 USD, when only 12% of participants earned more than $165 USD per month. There were many comments from respondents regarding fear and pain of multiple weekly injections around every lesion and the stigma from possessing CL associated scars. The study provides a unique and timely insight into the basis for targeted interventions aimed at improving health burden of CL for people in NWS through integrated interventions.

Echinococcus granulosus poses a major threat to human and animal health. Mu-class glutathione S-transferases (GSTs) are important detoxification enzymes in helminths and may participate in parasite-host interactions. In our previous proteomic analysis of extracellular vesicle (EV) preparations, E. granulosus Mu-class GST (Eg.GST-Mu) was identified in preparations derived from plasma collected from mice at 20 weeks post-infection and from hydatid cyst fluid. In the present study, recombinant Eg.GST-Mu (rEg.GST-Mu) was expressed and biochemically characterized, and the immunological responses associated with rEg.GST-Mu immunization in mice were preliminarily evaluated. The Eg.GST-Mu gene was cloned into the pET28a(+) vector and expressed in Escherichia coli BL21(DE3). The purified recombinant protein was verified using an anti-His antibody and rEg.GST-Mu-specific antiserum and retained detectable GST catalytic activity after refolding. Male C57BL/6 J mice were subsequently immunized with rEg.GST-Mu, and immune responses were evaluated by measuring antigen-specific IgG and its subclasses IgG1 and IgG2c, together with flow-cytometric analysis of splenic CD4[+] T-cell subsets. rEg.GST-Mu immunization elicited a robust antigen-specific humoral response and was associated with changes in the distribution of splenic CD4[+] T-cell subsets and the Treg/Th17 ratio. In vitro, rEg.GST-Mu attenuated the proliferative expansion of activated CD4[+] T cells. These findings demonstrate that rEg.GST-Mu possesses detectable GST catalytic activity and immunogenicity and provide an experimental basis for further investigation of its immunological functions. However, the role of native Eg.GST-Mu during natural E. granulosus infection, its potential association with EVs, and the mechanisms underlying its effects on host immune responses remain to be directly established.