Parasite standard strains are fundamental resources for parasitological research, disease control, and biopharmaceutical innovation. To address the incomplete system and lagging standardization of parasite strains in China, the National Parasite Resource Center organized 54 experts from all over the country to reach an important consensus after discussion, and the consensus system expounded the standardized rules for the screening, identification, preservation and application of parasite standard strains. The consensus clarified that parasite standard strains should have representativeness, genetic stability, and purity unity, which can be identified and quality controlled through multi-dimensional technologies such as morphology and molecular biology. Standardize distribution processes and application specifications in different scientific research scenarios, emphasizing the importance of ethical review, biosecurity, and resource benefit sharing. Suggestions are put forward to strengthen top-level design, improve the preservation system, promote the application of artificial intelligence (AI) technology, and establish a multidisciplinary collaboration platform to accelerate the construction of China’s parasite reference strain system in line with international standards, and enhance the national biological resource reserve capacity and scientific research competitiveness.

Objective To investigate the effects of BIX-01294, a histone H3 methyltransferase inhibitor, on the viability of protoscoleces and metacestode vesicles of Echinococcus granulosus. Methods Liver samples were collected from E. granulosus-infected sheep in local slaughterhouses. E. granulosus protoscoleces were isolated under sterile conditions and exposed to BIX-01294 solutions (dissolved in DMSO) at concentrations of 1, 5, 10, 25, and 50 µmol/L, while a solvent control group (DMSO group) served as controls. Protoscoleces were collected at 3, 6, 9, 12, and 15 days post-exposure and stained with methylene blue to observe viability. The survival rate of protoscoleces was calculated, and a survival curve was plotted to estimate the area under the survival curve (AUC) was determined using the trapezoidal rule. Protoscoleces were collected following exposure to 25 µmol/L BIX-01294 for 12 days and stained with hematoxylin and eosin (HE) to observe post-exposure changes. Protoscoleces exposed to 1 and 25 µmol/L BIX-01294 for 1, 6, 9, and 12 days were collected and stained with Caspase-3/7 dyes to assess the apoptosis of protoscoleces. The expression of H3K9me2 was determined using Western blotting in protoscoleces exposed to 25 µmol/L BIX-01294 for 12 days. The morphology of vesicles was observed under a microscope following exposure to 25 µmol/L BIX-01294 in vitro for 1, 3, 6, 9, and 12 days, and ultrastructural changes in vesicles were examined using scanning electron microscopy following exposure to BIX-01294 for 9 days. Results The viability of protoscoleces was (36.45 ± 7.14)% on day 6 post-treatment with 25 µmol/L BIX-01294, which significantly reduced as compared with that [(90.21 ± 3.72)%] on day 3 post-treatment (t = 15.86, P < 0.01), and the viability of protoscoleces was (71.91 ± 7.92)% on day 3 post-treatment with 50 µmol/L BIX-01294, which significantly reduced as compared with that (100%) on day 0 post-treatment (t = 6.25, P < 0.01), while the viability was (5.64 ± 1.48)% on day 6 post-treatment with 50 µmol/L BIX-01294, which continued to reduce as compared with that on day 3 post-treatment (t = 15.89, P < 0.01). The AUC values of BIX-01294 decreased in a dose-dependent manner with increasing concentrations, with the lowest AUC seen in the 50 µmol/L BIX-01294 treatment group (395.25%·d). Microscopy displayed shrinkage of protoscoleces, shedding of rostellum and hooklets, reduction of calcium granules and decreased motility following treatment with 50 µmol/L BIX-01294 for 6 days, and HE staining revealed loose, cavitary and irregular protoscoleces with a reduced volume, and thinned and incomplete body wall following treatment with 25 µmol/L BIX-01294 for 12 days. The highest Caspase-3/7 fluorescence intensity was recorded in protoscoleces exposed to 25 µmol/L BIX-01294 for 12 days. Western blotting assay determined lower H3K9me2 expression in protoscoleces treated with BIX-01294 for 12 days than in the DMSO group [(0.53 ± 0.08) vs. (1.00 ± 0.17); t = 4.226, P < 0.05]. Collapse or shrinkage of vesicles was found since day 6 following treatment with 25 µmol/L BIX-01294. Scanning electron microscopy displayed remarkable ultrastructural alterations, separation of germinal layer and cuticle and disorganization of the germinal layer structure in vesicles 9 days post-treatment with BIX-01294. Conclusion BIX-01294 exhibits a remarkable in vitro activity against E. granulosus protoscoleces and vesicles in a time- and concentration-dependent manner, accompanied by reduced H3K9me2 expression and upregulation of apoptosis-related signals, indicating that BIX-01294 has a potential value against echinococcosis.

Objective To develop a bidirectional developmental model for the differentiation of Echinococcus multilocularis protoscoleces into vesicles and for the development of somite, decipher the transcriptomic characteristics, and screen hub regulatory molecules. Methods E. multilocularis protoscoleces were induced into vesicle differentiation and somite development using an in vitro culture system. The early differentiation stage and the stable structure-formation stage were selected as representative time points, and changes in the parasite ultrastructure were observed using scanning electron microscopy. The mature stage of vesicle differentiation and the stage of pseudosomite formation were used as developmental time points. RNA was extracted from parasites for transcriptome sequencing, and differential expression genes (DEGs) were screened with protoscoleces as controls. In addition, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed, and protein-protein interaction (PPI) network was created. Results Swelling of the parasite margin and the visible vesicles were observed on day 9 of differentiation into vesicles, and the protoscolex rostellum degenerated with a reduction in microvilli on day 30 of differentiation into vesicles, with a vesicle formation rate of (34.2 ± 3.4)%. Transcriptomic analysis identified 3 910 DEGs on day 30 of differentiation into vesicles (60.15% of downregulated genes), which was significantly more than that (43.79% of downregulated genes, 621/1 418) on day 9 (χ² = 112.92, P < 0.01). These genes were mainly enriched in membrane receptor signaling and transmembrane transport pathways, and the hub genes were primarily involved in protein synthesis and mitochondrial energy metabolism. Evagination and active motility of protoscoleces was observed on day 3 of development into somite, while obvious neck constriction and posterior pseudosomite enlargement were seen on day 15, with a pseudosomite formation rate of (33.8 ± 2.9)%. Transcriptomic analysis identified 1 520 DEGs (57.83% of upregulated genes) on day 15, which was significantly more than that (51.97% of upregulated genes, 408/785) on day 3 (χ² = 7.19, P < 0.01). These genes were mainly enriched in cell cycle and external factor signaling pathways, and the hub genes were predominantly cell cycle regulatory molecules. Conclusion The bidirectional development of E. multilocularis protoscoleces is regulated by distinct transcriptional programs. Vesicle differentiation is characterized by metabolic and membrane system modulation, whereas somite development is dominated by cell cycle and Wnt signaling, deciphering the molecular regulatory patterns underlying bidirectional development of E. multilocularis protoscoleces. These findings provide the evidence for decoding the regulatory mechanisms of Echinococcus development and for screening potential targets.

Objective To evaluate the effectiveness of integrated echinococcosis control programmes in Ningxia, so as to provide insights into formulation of echinococcosis control measures. Methods Village-wide screening of human echinococcosis was performed in human screening points with a previous report of echinococcosis cases or positive Echinococcus coproantigens in domestic dogs from 19 cystic echinococcosis-endemic counties (cities, districts) in Ningxia from 2016 to 2025, and permanent residents at ages of 3 years and older received abdominal ultrasound scans for screening of echinococcosis. Fifty domestic dogs were sampled in each village (community), and the prevalence of Echinococcus coproantigens was detected using ELISA. One thousand sheep or 500 cattle at ages of one year and older from slaughterhouses were sampled for detection of Echinococcus infections in the visceras and the prevalence of Echinococcus infection was calculated in cattle and sheep. Four hundred mouse traps were deployed around three alveolar echinococcosis-endemic areas, and the prevalence of Echinococcus infection in the visceras was calculated in small rodents. Medical records were established for echinococcosis patients, and the efficacy of chemotherapeutic and surgical treatments was assessed. In addition, the awareness of human echinococcosis control knowledge was surveyed. Results The detection rate of cystic echinococcosis reduced from 1.60‰ (133/83 240) in 2016 to 0.36‰ (35/98 263) in 2025 (χ²_trend = 265.704, P < 0.05), and the detection rate of alveolar echinococcosis decreased from 0.20‰ (17/83 240) in 2016 to 0.01‰ (1/98 263) (χ²_trend = 45.581, P < 0.05). The overall prevalence of Echinococcus coproantigens decreased from 3.47% (547/15 776) in 2016 to 0.35% (50/14 290) in 2025 in domestic dogs (χ²_trend = 1 119.418, P < 0.05); however, there was a mild rebound in the prevalence in 2022 (0.75%, 106/14 173) and 2023 (0.76%, 167/21 916). The prevalence of Echinococcus infections appeared an overall tendency towards a decline in cattle (χ²_trend = 44.725, P < 0.05) and sheep (χ²_trend = 392.069, P < 0.05), with no infections detected in cattle in 2019, 2024 or 2025, and a reduction in the prevalence of Echinococcus infections in sheep by 1% in 2025 (1.07%, 78/7 291) relative to in 2016 (0.07%, 6/9 218). The prevalence of E. multilocularis infection was 0.19% (29/15 125) in rodents, with no infection detected in mice in 2020, 2021 or 2025; however, the prevalence rebounded from 2022 to 2024 (χ²_trend = 20.880, P < 0.05). The proportion of humans that were unwilling to receive treatments reduced by 4.77% in 2025 relative to in 2016, and the percentage of cure and improvements by chemical treatment increased by 15.42% in 2025 as compared by in 2016, while the recurrence rate of echinococcosis following surgical treatments reduced by 12.82% during the 10-year period. In addition, the awareness of human echinococcosis control knowledge increased from 56.04% (5 506/9 825) in 2016 to 86.49% (5 642/6 523) in 2025. Conclusion The integrated echinococcosis control programmes had achieved remarkable effectiveness in Ningxia from 2016 to 2025; however, the transmission cycle remains, and the cure rate of echinococcosis patients remains low. Future control efforts should focus on effective interruption of the transmission chain and precise medical cure of echinococcosis patients.

Objective To propose a 4-type ultrasound classification system based on the pathological features of hepatic alveolar echinococcosis (HAE), and to explore its consistency with pathological classification and clinical application value. Methods A retrospective analysis was performed on patients who were pathologically diagnosed with HAE after surgery at the First Affiliated Hospital of Xinjiang Medical University from January 2018 to December 2024. The ultrasound data, pathological data and treatment modalities of the patients were collected. Ultrasound examinations were conducted to record the location, maximum diameter, internal echo of the lesion, liquefactive necrosis area and calcification range. Referring to the ultrasound results and postoperative pathological features of the lesions, the lesions were divided into 4 types: infiltrative type, calcified type, liquefied type and complex type. The inter-observer consistency of ultrasound classification and the consistency between ultrasound classification and pathological results were comparatively analyzed, and the consistency evaluation was performed using the Kappa value. The differences in ultrasound-related parameters among the 4 types of lesions were compared, including the maximum diameter, the proportion of solid area, the proportion of calcified area, the proportion of liquefied area, and the hypoechoic halo around the lesion. The patient type was determined based on the classification of the largest lesion of the patient, and the clinical treatment measures of the patients were collected simultaneously to analyze the correlation between patient types and treatment modalities. Statistical analysis was carried out using SPSS 26.0 software. Results A total of 344 HAE patients and 469 HAE lesions were finally included. The ultrasound classification results showed 227 infiltrative lesions, 73 calcified lesions, 106 liquefied lesions and 63 complex lesions; the pathological classification results showed 217 infiltrative lesions, 81 calcified lesions, 110 liquefied lesions and 61 complex lesions. The overall coincidence rate of ultrasound classification was 94.5% (443/469), and the coincidence rates of each type of lesion were 100% (73/73) for calcified type, 97.2% (103/106) for liquefied type, 92.5% (210/227) for infiltrative type, and 90.5% (57/63) for complex type, respectively. The inter-observer consistency Kappa value was 0.936, and the Kappa value for the consistency between ultrasound classification and pathological results was 0.918. The differences in the proportions of solid, calcified, liquefied components and the maximum diameter among the 4 types of lesions were all statistically significant (H = 381.729, 237.657, 384.372, 101.102, all P < 0.05). 76.7% (264/344) of the patients underwent radical resection for HAE, 21.2% (73/344) underwent autologous liver transplantation, and 2.0% (7/344) underwent allogeneic liver transplantation. Preoperatively, 12.8% (44/344) of the patients received palliative treatment, including 16 cases of percutaneous transhepatic biliary decompression, and 25 cases of liquefied type and 3 cases of complex type underwent puncture drainage of the abscess cavity. The proportions of patients with different types were 44.5% (153/344) for infiltrative type, 12.5% (43/344) for calcified type, 27.3% (94/344) for liquefied type, and 15.7% (54/344) for complex type. Correlation analysis showed that there was a statistically significant difference in the selection rate of abscess cavity puncture and drainage among different patient types (χ² = 47.32, P < 0.01). Although the difference in the selection rate of radical lesion resection among different patient types was statistically significant (χ² = 10.47, P < 0.05), after Bonferroni correction (α = 0.008 33), there was no statistically significant difference in the selection rate between any two types. Conclusion This HAE ultrasound classification is highly consistent with pathological features and demonstrates high reproducibility, with each subtype possessing characteristic ultrasound parameters. This classification method, by identifying the main components of the lesion preoperatively, provides the imaging basis for the individualized treatment of hepatic alveolar echinococcosis.

Objective To investigate the prevalence of Echinococcus infections in canines and the risk factors and behaviors for human echinococcosis in Guoluo Tibetan Autonomous Prefecture, Qinghai Province, so as to provide insights into formulation of precise echinococcosis control in this region. Methods Pastoral dog-raising households were randomly sampled from 11 townships of Banma County, Dari County and Jiuzhi County in Guoluo for surveying on risk factors for domestic dog infection using a random sampling method in 2025, and fresh or apparently intact fecal samples were collected from domestic dogs and wild canines. A multiplex quantitative Real-time PCR (qPCR) assay was employed to detect Echinococcus infection status in fecal samples. Based on the previous surveys, the risk factors and behaviors of human echinococcosis were examined in Bama County and Dari County, Guoluo in 2025. Univariate analysis was performed with a history of human echinococcosis as an dependent variable, and a logistic regression model was created to identify independent factors affecting the development of human echinococcosis. All statistical analyses were performed with the statistical software SPSS 25.0, and chi-square test or Fisher’s exact test was used for comparison of constituent ratios. Results A total of 945 canine fecal samples were collected, including 696 fecal samples from domestic dogs and 249 fecal samples from wild canines. The overall prevalence of Echinococcus was 1.59% (15/945) in dog feces, and the detection of Echinococcus was higher in fecal samples from domestic dogs than those from wild dogs [0.43% (3/696) vs. 4.82% (12/249); χ² = 19.885, P < 0.01]. A total of 459 questionnaires on risk factors for Echinococcus infection in domestic dogs were collected. The analysis showed that the fecal positivity rate in domestic dogs differed significantly among different tying methods(χ² = 8.400, P < 0.05), while no statistical significance was found for the other risk factors (P > 0.05). A total of 400 questionnaires pertaining to human echinococcosis behaviors, and 346 valid questionnaires were recovered, with a recovery rate of 86.50%. The awareness and qualification rates of echinococcosis control knowledge were 86.12% (1 788/2 076) and 86.99% (301/346). A significant difference was found in the qualification rate of human echinococcosis knowledge between Banma County (96.95%, 159/164) and Dari County (78.02%, 142/182) (χ² = 25.672, P < 0.05), and significant differences were detected in the qualification rate among individuals with dog breeding at home, presence of stray dogs or wild canines near the residence, behaviors of lying on grass or biting grass, and household disposal of raw bovine and goat viscera (χ² = 18.155, 5.115, 3.958, 32.647; all P < 0.05). The univariate analysis results showed that there was a significant difference in the prevalence of human echinococcosis between Banma County and Dari County (χ² = 4.984, P < 0.05), and significant differences were found in the prevalence of human echinococcosis in terms of populations, age groups, education level, dog breeding at home, presence of stray dogs or wild canines near the residence, and behaviors of lying on grass or biting grass (χ² = 9.298, 14.202, 10.426, 8.082, 7.247, all P < 0.05). Multivariate logistic regression analysis identified age group (OR = 2.075, 95% CI: 1.041 to 4.138), education level (OR = 0.440, 95% CI: 0.224 to 0.865), dog breeding at home (OR = 2.482, 95% CI: 1.017 to 6.057), and presence of stray dogs or wild canines near the residence (OR = 3.094, 95% CI: 1.096 to 8.732) as independent factors affecting the prevalence of human echinococcosis. Conclusion The management of Echinococcus infections is satisfactory in domestic dogs raised by pastoral households in Guoluo Prefecture, Qinghai Province; however, there are persistent risk factors for human Echinococcus infections, which requires reinforced health education for pastoral households and standardized management for canine breeding. Given the high prevalence of Echinococcus infections in wild canines, the control system targeting wild canines needs to be optimized to consolidate the gained echinococcosis control achievements.

Objective To investigate and analyze the prevalence and changing trends of major human parasitic diseases in Jiangxi Province, so as to provide insights into optimization of the control strategies and measures. Methods According to the Protocol for National Survey on the Status of Major Human Parasitic Diseases in China in 2025 and based on the actual conditions of Jiangxi Province, soil-transmitted helminth infections were surveyed in ecological zones using a multistage stratified cluster sampling method according to economic conditions and geographical locations. Each survey county was divided into five regions (northeastern, southeastern, southwestern, northwestern, and central) according to geographical locations, and one township was randomly selected from each region. Then, one natural village was randomly selected from each township as the survey point, with 200 individuals surveyed per point. Three townships were randomly sampled from Xinfeng County, an area endemic for Clonorchis sinensis in Jiangxi Province, and one natural village was randomly sampled from each township as the survey point, with 200 individuals surveyed per point. Helminth (soil-transmitted nematodes, C. sinensis and Taenia spp.) eggs were detected using a modified Kato-Katz thick smear technique, while pinworms were detected among children at ages of 3 to 9 years using a cellophane tape test. Results A total of 18 155 individuals were surveyed across 88 survey points in 18 counties of Jiangxi Province, including 896 children detected for pinworm infections. Six species of intestinal helminths were identified, including hookworm, Trichuris trichiura, Enterobius vermicularis, C. sinensis, Paragonimus, and Echinostoma. The average prevalence rates of intestinal helminth infections and soil-transmitted nematode infections were 1.47% (266/18 155) and 0.90% (163/18 155), while the prevalence rates of hookworm, T. trichiura, E. vermicularis, C. sinensis infections were 0.75% (137/18 155), 0.15% (27/18 155), 0.11% (20/18 155), and 0.44% (80/18 155), respectively. In addition, Echinostoma infections were detected among 5 individuals and Paragonimus infection occurred among one individual. The prevalence of pinworm infections was 2.01% (18/896) among children at ages of 3 to 9 years. The highest prevalence of helminth infections was recorded in the Nanling Mountain and Hilly Ecological Zone (2.96%, 79/2 666), while the lowest was seen in the Zhejiang-Fujian Mountain and Hilly Ecological Zone (0.34%, 7/2 035) (χ² = 44.09, P < 0.01). The highest prevalence of soil-transmitted helminth infections (1.38%, 101/7 324) was found in the Hunan-Jiangxi Hilly and Mountain Ecological Zone (χ² = 37.97, P < 0.01), with the highest prevalence of T. trichiura infections in the Plain Ecological Zone of the Middle and Lower reaches of the Yangtze River (0.28%, 17/6 130), the highest prevalence of hookworm infections in the Hunan-Jiangxi Hilly and Mountain Ecological Zone (1.27%, 93/7 324) (χ² = 44.91, P < 0.01), and 88.75% (71/80) of C. sinensis infections concentrated in the Nanling Mountain and Hilly Ecological Zone. In terms of survey counties, the highest prevalence of hookworm infections was seen in Le’an County (2.19%, 23/1 052), and the highest prevalence of C. sinensis infections was recorded in Xinfeng County (11.61%, 70/603). In terms of genders, the prevalence of C. sinensis infections was higher among men (0.72%, 63/8 799) than among women (0.18%, 17/9 356) (χ² = 29.51, P < 0.01), and the prevalence of soil-transmitted nematode infections was higher among women (1.10%, 103/9 356) than among men (0.68%, 60/8 799) (χ² = 8.95, P < 0.05). Helminth infections were detected among individuals across all age groups, with the highest prevalence found among individuals at ages of 70 years and older (2.27%, 110/4 849) and the lowest at ages of 10 to 19 years (0.41%, 6/1 446). Parasitic infections recorded among individuals with all occupations except scattered children, and the highest prevalence rates of helminth (1.70%, 227/13 354), soil-transmitted nematode (1.12%, 150/13 354), T. trichiura (0.18%, 24/13 354), hookworm (0.95%, 127/13 354) and C. sinensis infections (0.55%, 74/13 354) were all found among farmers. Conclusion The infection rates of key parasites among the population in Jiangxi Province have all dropped to historical lows, yet certain issues still stand out prominently-such as hookworm infections among people aged 60 and above, pinworm infections in children aged 3 to 9, and C. sinensis infections in regions where raw freshwater fish is consumed. The health education-led integrated control strategy with an emphasis on the management of the source of infections has achieved remarkable effectiveness, which is recommended to be implemented for a long time of period.

Objective To investigate the prevalence of human soil-transmitted helminth infections in Chongqing Municipality from 2017 to 2024, so as to provide insights into formulation of soil-transmitted helminthiasis control strategies and optimization and adjustment of soil-transmitted helminthiasis control measures. Methods Several counties (districts) were selected from Chongqing Municipality annually during the period from 2017 to 2024 as surveillance points, according to the requirements of the National Surveillance Protocol for Clonorchiasis and Soil-Transmitted Helminthiases (Trial). Each surveillance county was divided into five geographical regions (eastern, western, southern, northern, and central), and one administrative village from one township was randomly selected from each region for surveillance programs, with at least 200 individuals surveyed in each village, and at least 1 000 individuals surveyed in each surveillance county. Stool samples were collected from participants, and soil-transmitted helminth eggs were detected using the modified Kato-Katz thick smear technique (two slides per stool sample). In addition, the hookworm species was identified with the test-tube filter-paper cultivation technique, and the prevalence and intensity of infections with hookworm species were calculated. Differences of proportions were tested for statistical significance with chi-square test. Results A total of 61 430 residents were detected for soil-transmitted helminth infections from 2017 to 2024, with an overall prevalence of 3.05% (1 875/61 430), and the prevalence of hookworm, Ascaris lumbricoides, and Trichuris trichiura were 2.66% (1 636/61 430), 0.36% (220/61 430), and 0.06% (35/61 430), respectively. Hookworm larvae were successfully identified in 777 stool samples, including 500 samples identified with Necator americanus and 277 with Ancylostoma duodenale. The annual prevalence rates of human soil-transmitted helminth infections were 6.91% (424/6 132), 5.75% (354/6 157), 3.87% (238/6 149), 1.23% (76/6 155), 2.89% (236/8 159), 0.50% (56/11 093), 2.84% (258/9 069), and 2.74% (233/8 516) in Chongqing Municipality from 2017 to 2024, respectively (χ² = 791.302, P < 0.01). The highest prevalence of soil-transmitted helminth infections was seen in the western Chongqing areas (4.63%, 577/12 466), and the lowest recorded in central urban areas (0.89%, 161/18 179). In terms of gender distribution, the prevalence of soil-transmitted helminth infections was 3.02% (873/28 881) among men and 3.08% (1 002/32 549) among women (χ² = 0.160, P > 0.05). Of participants, the highest prevalence of soil-transmitted helminth infections was detected among residents at ages of 70 years and older by age groups (6.01%, 681/11 337), among farmers by occupations (4.77%, 1 699/35 641), and among illiterate or semi-literate individuals by educational levels (5.08%, 206/4 056). There are significant differences in the prevalence of human soil-transmitted helminth infections in terms of regions, age groups, occupations, and education levels (χ² = 535.934, 833.397, 850.287, and 534.637, respectively; all P values < 0.05). Mild infections predominated among individuals with hookworm, A. lumbricoides, and T. trichiura infections, and the proportions of mild, moderate, and severe infections were 92.30% (1 510/1 636), 4.28% (70/1 636), and 3.42% (56/1 636) among individuals with hookworm infections, respectively. The highest infection intensity was 65 520 among individuals with hookworm infections, 297 792 among individuals with A. lumbricoides, and 2 568 among individuals with T. trichiura infections. Conclusion The overall prevalence of soil-transmitted helminth infections appeared an overall tendency towards a decline in Chongqing Municipality from 2017 to 2024; however, the prevalence remained high in some districts (counties) of western Chongqing areas. Severe hookworm infections were concentrated among the elderly residents. Improved control efforts and health education are required targeting high-prevalence areas and high-risk populations.

Objective To investigate the in vitro killing effect of halofuginone (Hal) on Echinococcus multilocularis and Trichinella spiralis. Methods E. multilocularis protoscoleces and cysts were isolated from mice infected with E. multilocularis, and muscle larvae, adult worms and newborn larvae of T. spiralis were isolated from mice infected with T. spiralis, followed by in vitro culture in DMEM medium. E. multilocularis protoscolices were treated with Hal at final concentrations of 50, 80, 100 μmol/L, 1% dimethyl sulfoxide (DMSO) and equal volume of medium respectively. After 24, 48 and 72 hours of intervention, the morphology of protoscolices was observed, 0.1% methylene blue staining was used to calculate the mortality of protoscolices in each group, and the half-maximal effective concentration (EC₅₀) of Hal against protoscolices was determined. E. multilocularis cysts were treated with Hal at a final concentration of 100 μmol/L and 1% DMSO respectively, and the ultrastructure of cysts was observed by scanning electron microscopy after 72 hours of intervention. The muscle larvae, adult worms and newborn larvae of T. spiralis were treated with Hal at final concentrations of 50, 80, 100 μmol/L, 1% DMSO and equal volume of medium respectively. The morphology and viability of muscle larvae and adult worms were observed after 24, 48 and 72 hours of intervention, and the morphology and viability of newborn larvae were observed after 96, 120 and 144 hours of intervention. Statistical analysis was performed using GraphPad Prism 9.5, and one-way ANOVA was adopted for data comparison. Results After 48 hours of intervention, the mortality rates of E. multilocularis protoscolices in the 50, 80 and 100 μmol/L Hal groups were (13.05 ± 1.43)%, (57.57 ± 2.57)% and (96.38 ± 2.16)% respectively, all significantly higher than (1.08 ± 0.52)% in the DMSO group (t = 13.587, 37.254, 74.430, all P < 0.01), with an EC₅₀ of 73.96 μmol/L. After 72 hours of intervention with 100 μmol/L Hal, the protoscolices of E. multilocularis showed structural damages including body surface rupture, disordered microvilli arrangement and destroyed suckers under scanning electron microscopy; irregular cracks and pores appeared on the surface of the cyst tegument of E. multilocularis, the germinal layer was extensively exfoliated, and the number of immature protoscolices was significantly reduced. After 72 hours of intervention, the mortality rates of T. spiralis muscle larvae in the 50, 80 and 100 μmol/L Hal groups were (83.59 ± 5.76)%, (87.57 ± 1.52)% and (95.42 ± 1.21)% respectively, all significantly higher than (1.58 ± 0.38)% in the DMSO group (t = 24.590, 94.801, 127.887, all P < 0.01); the mortality rates of adult worms in the 50, 80 and 100 μmol/L Hal groups were (9.32 ± 0.96)%, (36.90 ± 3.08)% and (98.4 ± 1.42)% respectively, all significantly higher than (1.77 ± 0.58)% in the DMSO group (t = 13.201, 19.698, 115.971, all P < 0.01). After 120 hours of intervention, the mortality rates of newborn larvae in the 50, 80 and 100 μmol/L Hal groups were (89.11 ± 1.40)%, (92.32 ± 1.17)% and (95.31 ± 2.31)% respectively, all significantly higher than (1.68 ± 0.16)% in the DMSO group (t = 107.394, 133.302, 70.129, all P < 0.01). After Hal intervention, all T. spiralis muscle larvae, adult worms and newborn larvae showed typical rigid dead morphology without autonomous movement. Conclusion Hal exerts highly efficient anthelmintic effect by destroying the structure of parasites in vitro, which is expected to be a potential drug for the treatment of echinococcosis multilocularis and trichinellosis.

Objective To explore the immunoprotective effect and mechanism of exosomes derived from Plasmodium-infected red blood cells on experimental cerebral malaria (ECM) in mice. Methods Exosomes were extracted and identified from peripheral blood of BALB/c mice infected with Plasmodium yoelii 17XNL (P.y17XNL). C57BL/6J mice were randomly divided into ECM group, exosome immunization group (Exos + ECM group) and negative control group (NC group). Mice in the Exos + ECM group received two doses of exosome immunization via tail vein injection; 20 days after immunization, mice in the ECM group and Exos + ECM group were intraperitoneally injected with 1 × 10⁶ P. berghei ANKA (P.bANKA)-infected red blood cells, while mice in the NC group were intraperitoneally injected with an equal volume of normal saline. The erythrocyte infection rate and mice survival rate were dynamically observed. Mice brain tissues were collected, and the absorbance at 630 nm (A₆₃₀ value) of the supernatant was detected after Evans blue perfusion to evaluate the integrity of the blood-brain barrier in mice. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of mice brain tissue, and immunohistochemical staining was performed to detect the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in brain microvessels. Western blotting and real-time fluorescent quantitative reverse transcription PCR (qRT-PCR) were used to detect the changes of inflammation-related cytokines in mice brain tissue. Proteomics analysis was combined to explore the regulatory association between core differential components of exosomes and brain tissue inflammatory factors. Results The erythrocyte infection rates of the ECM group and Exos + ECM group reached the peak on day 11 and day 15 post-infection, with values of (25.0 ± 0.0)% and (17.2 ± 4.4)%, respectively, and the erythrocyte infection rate in the Exos + ECM group increased more slowly. All mice in the ECM group died of cerebral malaria on day 13 post-infection, and the survival rate of mice in the Exos + ECM group was 83.3% on day 15 post-infection. The detection results of blood-brain barrier integrity showed that the blood-brain barrier in the ECM group was damaged, while the damage in the Exos + ECM group was not obvious. the A₆₃₀ values of the supernatant in the NC group and Exos + ECM group were (0.28 ± 0.04) and (0.62 ± 0.01) respectively, both lower than (0.89 ± 0.02) in the ECM group (t = 24.24, 10.53, both P < 0.01). The results of HE staining and immunohistochemistry of brain tissue showed that, compared with the ECM group, the number of infected red blood cells in brain microvessels, inflammatory infiltration, and the expression levels of ICAM-1 and VCAM-1 in the Exos + ECM group were reduced. The qRT-PCR results showed that the relative mRNA transcription levels of ICAM-1, VCAM-1, interleukin-6 (IL-6), IL-17A, transforming growth factor-β (TGF-β) and IL-10 in the brain of Exos + ECM group mice were 4.05 ± 0.42, 1.24 ± 0.14, 0.48 ± 0.3, 0.27 ± 0.31, 1.39 ± 0.53 and 4.23 ± 3.29, respectively, which were lower than 8.11 ± 0.86, 2.80 ± 0.33, 1.38 ± 0.76, 2.28 ± 0.26, 2.94 ± 0.49 and 16.02 ± 3.56 in the ECM group (t = 8.76, 8.52, 2.91, 6.81, 4.51, 5.41, P < 0.01, 0.01, 0.05, 0.01, 0.05, 0.01). Western blotting results showed that the protein expression levels of Toll-like receptor 4 (TLR4), IL-1β and tumor necrosis factor-α (TNF-α) in the brain of Exos + ECM group mice were 0.84 ± 0.06, 0.81 ± 0.04 and 2.07 ± 0.72, respectively, which were lower than 1.24 ± 0.04, 0.85 ± 0.03 and 6.06 ± 2.45 in the ECM group (t = 7.07, 1.27, 3.12, all P < 0.05). Proteomics results showed that the differential proteins of exosomes from P.y17XNL-infected mice and normal mice were mainly enriched in nuclear proteins, cytoplasmic proteins, plasma membrane proteins and mitochondrial proteins, which were related to protein export, transport and amino acid metabolism. Conclusion Exosome immunization had an immunoprotective effect on ECM, which can reduce the erythrocyte infection rate and improve the survival rate of mice. The main mechanism may be related to reducing brain inflammation, protecting the integrity of the blood-brain barrier and improving the pathological damage of mice brain tissue.

Objective To analyze the trends in relapse of Plasmodium vivax malaria in Dehong, Yunnan from 2014 to 2025, so as to provide insights into formulation of malaria control strategies. Methods A retrospective cross-sectional study was performed to analyze the trend of relapse rate of Plasmodium vivax. All data pertaining to reported P. vivax malaria cases in Dehong from 2014 to 2025 were extracted from the China Disease Control and Prevention Information System. Relapsed cases were verified with the duplicate checking function, and the relapse rate was calculated. Shapiro-Wilk test was used to test for normality and Levene’s test was used to test for homogeneity of variances. Independent-sample t-test was employed to compare the differences in relapse rates during the period between 2014-2019 and 2020-2025. In addition, the trends in the relapse rate were examined using interrupted time series (ITS) analysis with the intervention breakpoint of “switching to WHO-donated and Cyprus-produced primaquine for radical treatment of P. vivax and P. ovale malaria in Yunnan Province in 2020”. Results A total of 2 485 P. vivax malaria cases were reported in Dehong from 2014 to 2025, including 2 454 overseas imported cases (98.75%) and 79 relapsed cases, with an overall relapse rate of 3.18% (79/2 485). There was a significant difference in the relapse rate between 2020 and 2025 [(4.60 ± 1.82)%] and between 2014 and 2019 [(1.64 ± 1.40)%] (t = -3.158, P < 0.05), and an average increase of 2.96% [95% confidence interval (CI): 0.87% to 5.04%, P < 0.05] was seen in the relapse rate during the period from 2020 to 2025 relative to from 2014 to 2019. From 2014 to 2025, the vivax malaria relapse rate in Dehong fluctuated generally, and 2020 was an obvious breakpoint in the time series. ITS regression analysis revealed that the regression coefficients for the centering time variable, intervention variable, interaction term, and lagged variable were -0.363 (95% CI: -1.890 to 1.163; t = -0.582, P > 0.05), 5.115 (95% CI: -1.076 to 11.307; t = 2.021, P > 0.05), 0.078 (95% CI: -1.840 to 1.996; t = 0.099, P > 0.05), -0.069 (95% CI: -0.872 to 0.734; t = -0.210, P > 0.05), indicating a tendency towards a decline in the relapse rate of P. vivax malaria in Dehong during the period from 2014 to 2019 and from 2020 to 2025. In addition, the relapse rate exhibited an immediate increase at the intervention point in 2020, and the tendency towards a decline in the relapse rate slowed slightly after 2020. Conclusion The changing trends in the relapse rate of P. vivax malaria is not associated with the replacement of foreign countries-produced primaquine in Dehong. The relapse rate remains low, and there is no need to change the radical cure regimen. However, enhanced surveillance is required.

Objective To investigate the resistance to pyrethroid insecticides and metabolic enzyme levels in Aedes albopictus from Anshun City, Guizhou Province, and to analyze the genotypes and distribution characteristics of the knockdown resistance (kdr) gene, so as to provide a theoretical basis for the chemical control of Ae. albopictus. Methods The larvae of Ae. albopictus were captured from small water containers in residential areas at different locations of Anshun City from July 2023 to December 2024, and then reared in laboratory to F1 to F2 generations. The resistance to deltamethrin, beta-cypermethrin and permethrin was tested in late 3rd-instar and early 4th-instar mosquito larvae of the susceptible strain and field strain with the larval dipping method, and the resistance to the three pyrethroid insecticides was determined in non-blood-fed female adults of the field strain 3 to 5 days following emergence with the tube-test method. The activity of non-specific esterase (NSE), mixed function oxidase (MFO), and glutathione-S-transferase (GST) was measured with a microplate reader in larval mosquitoes without bioassay. Genomic DNA was extracted from individual adult mosquito after bioassay, and partial fragments of the voltage-gated sodium channel (vgsc) gene were amplified using PCR assay. Following sequencing, the sequences were aligned with reference sequences from NCBI to analyze the mutations and frequency distributions of loci V1016, I1532, F1534 and D1763, respectively. The degree of resistance to pyrethroid insecticides in larval mosquitoes was graded using the software SPSS 31.0 professional, and Shapiro-Wilk test and Wilcoxon rank-sum test were performed to determine the distribution of enzyme activity. In addition, the relative importance of metabolic mechanisms and kdr mechanisms in insecticide resistance in Ae. albopictus were predicted by the classification and regression trees (CART) algorithm. Results Larvae of Ae. albopictus exhibited resistance ratios of 38.47, 18.21 and 209.17-fold to deltamethrin, beta-cypermethrin and permethrin, respectively, all appearing a moderate to high resistance level. The mortality rates of adult mosquitoes were 2.04% (2/98), 2.06% (2/97)and 7.00% (7/100), respectively, indicating resistance to all three pyrethroid insecticides. The activities of NSE, MFO, and GST were 356.17 (321.65 to 497.79) nmol α-naphthol/(min·mg pr), 12.88 (11.17 to 18.01) nmol cyt c/mg pr, and 128.70 (116.63 to 153.03) nmol/(min·mg pr) in the field population of Ae. albopictus, respectively, all showing a positively skewed distribution, which were 1.26, 0.45, and 1.65 times higher than those in susceptible strains (Z = ‒4.030, ‒8.191 and ‒7.371, all P values < 0.01). kdr gene mutations were not detected at the locus 1763 in Ae. albopictus from Anshun City, and were detected at loci V1016, I1532, and F1534. At loci 1016 and 1532, wild-type homozygotes were predominant genotypes (65.97%, 98.61%), followed by wild-type/mutant heterozygotes (30.9%, 1.39%), with the lowest proportion seen for the mutant homozygotes (3.13%, 0). There were three alleles at locus 1534, including wild-type (TTC/F), mutant (TCC/S) and mutant (TGC/C), and the mutant-type (TCC/S) had a higher gene frequency than the wild-type (TTC/F). At locus 1534, the mutant homozygote (S/S) had the highest genotype frequency (61.11%), followed by the wild/mutant-type heterozygote (F/S) (28.47%) and wild-type homozygote (F/F) (7.64%), and the mutant homozygote (C/C) had the lowest genotype frequency (2.78%). A total of 10 genotype combinations were detected in the three-locus mutation test, and the T1 genotype (V/V + I/I + S/S) accounted for the highest proportion (60.76%, 175/288), followed by the T2 genotype (V/G + I/I + F/S) (25.69%, 74/288). Results from CART analysis showed that the factors contributing to pyrethroid resistance in Ae. albopictus in Anshun City included mutation at locus 1534 (100%), MFO activity (96.7%), GST activity (66.6%), NSE activity (21.4%), mutation at locus 1016 (16.4%), and mutation at locus 1532 (1.2%). Conclusion Ae. albopictus has developed resistance to pyrethroid insecticides at varying degrees in Anshun City. The NSE, MFO or GST activities have not yet presented typical characteristics of metabolic resistance, and kdr gene mutation is dominated at locus F1534. Emergency of resistance to pyrethroid insecticides in Ae. albopictus results from the synergistic effect of changes in metabolic detoxification enzyme activities and gene mutations.

Objective To compare the mosquito capturing effectiveness of an artifical intelligent (AI)-based mosquito surveillance method (the intelligent trap method) with three traditional vector surveillance methods, including the light trap method, the human-baited double net trap method and the BG-trap method, so as to provide insights into the application of AI-based intelligent systems in mosquito surveillance. Methods A residential community with more than 500 households and a high mosquito density was selected from Xiuzhou District, Jiaxing City. Mosquito surveillance was simultaneously performed with the light trap method, the human-baited double net trap method, and the BG-trap method one day per week during the period from May to September 2025, while the intelligent mosquito trap was run as a 24/7 service. The light trap index, mosquito biting index, mosquito attraction index and the adult mosquito density captured by intelligent traps during the same period were calculated. The mosquito-capturing effectiveness was compared between the intelligent trap method and three traditional mosquito monitoring methods with the Kruskal-Wallis H test, and the mosquito-capturing outcomes and trends in the mosquito density of these four mosquito monitoring methods, as well as the accuracy of the intelligent trap system for identification of mosquito species and genders were descriptively analyzed. Results A total of 4 620, 23, 250, and 3 564 adult mosquitoes were captured with intelligent traps, human-baited double net traps, BG-traps, and light traps during the monitoring period, including 299 (6.47% of total mosquitoes captured), 16 (69.57%), 247 (98.80%), and 88 Aedes albopictus (2.47%), respectively. At weekly monitoring days with three traditional monitoring methods, 2 female adult mosquitoes were captured with intelligent traps during daytime (15∶00-17∶00), with an average adult mosquito density of 0.05 mosquitoes per hour, and 219 female adults were captured during nighttime, with an average density of 9.95 mosquitoes per night, while a total of 21 female adults were captured with human-baited double net traps, with an average mosquito biting index of 0.95 mosquitoes per hour. In addition, a total of 150 female adults were captured with BG-traps, with an average mosquito attraction index of 6.82 mosquitoes per hour during the monitoring period, and a total of 1 104 female adults were captured with light traps, with an average light trap index of 25.09 mosquitoes per light per night. There was a significant difference in the average mosquito-capturing rate at the same period during daytime among the intelligent trap method (0.05 mosquitoes per hour), the human-baited double net trap method (0.95 mosquitoes per hour), and the BG-trap method (6.82 mosquitoes per hour) (H = 39.559, P < 0.05), with a lower average mosquito-capturing rate seen for the intelligent trap method than the human-baited double net trap method and the BG-trap method (both P < 0.05), and a lower average mosquito-capturing rate seen for the intelligent trap method than the light trap method at the same period during nighttime (9.95 mosquitoes per night vs. 25.09 mosquitoes per light per night; H = 6.377, P < 0.05). A similar trend was seen in the fluctuation of the adult mosquito density between the intelligent trap method and the light trap method, with both peaking in week 8 (June 24), at densities of 29 mosquitoes per night and 118.5 mosquitoes per light per night, respectively. In addition, the number of adult mosquitoes recorded in the backend database of the intelligent traps was higher than the manually collected count (4 620 vs. 3 339 adult mosquitoes) from May 1 to September 29, and the estimated accuracy of AI-based intelligent traps was 88.71% for identification of mosquito species and 90.03% for identification of mosquito genders. Conclusion Although the trend in fluctuation of the adult mosquito density is comparable between the intelligent trap method and the light trap method, its mosquito-capturing rate is significantly lower than those of three traditional methods, including the light trap method, human-baited double net trap method and the BG-trap method. In addition, the number of adult mosquitoes recorded in the backend database of intelligent traps differs from the manually collected count, which requires further optimizations.

Objective To explore the application architecture and effectiveness of implementation of an intelligent surveillance and early warning system for schistosomiasis based on the One Health concept in elimination areas, so as to provide insights into consolidation of schistosomiasis elimination achievements in similar areas. Methods The organizational structure of government-led and multi-sectoral collaborative surveillance and early warning for schistosomiasis was established in Changshan County, Zhejiang Province in 2024, and an intelligent surveillance and early warning system for schistosomiasis was run for integrated surveillance and early warning of human-animal-environment, which was consisted of four modules, including active surveillance of humans, surveillance of hospital visits, surveillance of host animals, and snail surveillance. Multi-source surveillance data were collected from the system from 2024 to 2025, and compared with historical baseline data. A total of indicators from 5 categories were collected, including transmission route, source of infection, control capability, population vulnerability, and natural factors, and the risk ranking of 10 administrative villages was calculated to evaluate the effectiveness in risk identification, early warning and response, and control measures. Results An intelligent multi-point triggered, government-led, multisector-coordinated, data-shared and intelligent-analyzed surveillance and early warning system for schistosomiasis was successfully established in Changshan County, which achieved all-weather, automated surveillance and data aggregation of human, livestock, host animals, snails, environments, and factors of imported risk. A total of 154 000 pieces of data pertaining to integration of medical services and prevention were automatically collected by the system, and a total of 25 303 person-time high-risk populations were screened. The time of early warning and response reduced by a mean period of 3 days compared to traditional modes, and the mean duration from data triggered to early warning information sent to administrators was shortened within 8 h. The efficiency of precise snail control increased by approximately 40% in key settings, and the coverage rate of livestock surveillance was 96.7%. A total of 11 imported risk events received successful early warning and response. Of 10 administrative villages, 4 had schistosomiasis transmission risk ranking of higher than 0.2, 5 between 0.1 and 0.2, and 1 below 0.1. Conclusion The intelligent surveillance and early warning system for schistosomiasis based on the One Health concept may effective integrate fragmented information and significantly enhance the capabilities of early detection, rapid assessment, and precise interventions for imported and re-emerging risks of schistosomiasis in elimination areas, which is an indispensable choice and a powerful tool for achieving and sustaining elimination.

Objective To systematically evaluate the anti-Toxoplasma gondii activity and safety of benzoyl metronidazole (BMZ). Methods Vero cells were treated with BMZ at different concentrations (50, 100, 200, 400, 500, 600, 800, 1 000 μg/mL), and the cytotoxicity of BMZ to Vero cells was detected by cell counting kit-8 (CCK-8) assay. After inoculation with Toxoplasma tachyzoites, Vero cells were treated with BMZ at the above concentrations, and the killing effect of BMZ on Toxoplasma was determined by CCK-8 assay. Vero cells inoculated with Toxoplasma tachyzoites were treated with 50 μg/mL BMZ, and the quantitative and morphological changes of Toxoplasma tachyzoites were observed via Giemsa staining and scanning electron microscopy. Kunming mice were randomly divided into low-dose BMZ group (LD-BMZ group), medium-dose BMZ group (MD-BMZ group), high-dose BMZ group (HD-BMZ group), sulfadiazine sodium group (SD-Na group) and negative control group (NC group). All mice were intraperitoneally inoculated with 1 × 10⁴ Toxoplasma tachyzoites per mL. Four hours after infection, mice were intragastrically administered continuously for 5 days: the LD-BMZ, MD-BMZ and HD-BMZ groups received BMZ at final doses of 12.5, 25 and 50 mg/kg, respectively, the SD-Na group received SD-Na at a final dose of 50 mg/kg, and the NC group received an equal volume of corn oil. On day 7 post-infection, the survival rates of mice were recorded, and the parasite load in mice ascites was counted using a hemocytometer. The heart, liver, spleen, lung, kidney and brain tissues of mice in the MD-BMZ group and NC group were collected. After DNA extraction, qPCR was performed to detect the parasite load in each tissue, and the tissues were processed into pathological sections for HE staining to observe the histopathological changes. GraphPad Prism 9 software was used for data analysis and plotting, and independent sample t-test or one-way ANOVA was adopted for significance analysis. Results The cytotoxicity assay showed that the CC₅₀ of BMZ to Vero cells was 155.67 μg/mL; the cell viability was (108.78 ± 1.00)% when BMZ concentration was 50 μg/mL, and decreased to (81.22 ± 3.34)% at 100 μg/mL. Therefore, 50 μg/mL was determined as the maximum safe concentration of BMZ. The in vitro anti-parasitic assay showed that BMZ could inhibit the proliferation of intracellular Toxoplasma tachyzoites, and the inhibition rate increased with the elevation of BMZ concentration; the inhibition rate of BMZ against Toxoplasma reached 14.3% at 50 μg/mL. Giemsa staining results showed that the number of Toxoplasma tachyzoites in cells of the BMZ group was reduced. Scanning electron microscopy observation showed that normal Toxoplasma tachyzoites presented typical spindle or crescent structures with complete contours, plump morphology and smooth surface; while after BMZ treatment, the structural integrity of Toxoplasma was completely lost, with blurred contours and destroyed cell membrane. The in vivo anti-parasitic assay showed that on day 7 post Toxoplasma infection, the survival rates of mice in the LD-BMZ, MD-BMZ, HD-BMZ, SD-Na and NC groups were 2/10, 3/10, 2/10, 2/10 and 1/10, respectively, with no statistically significant difference (χ² = 3.19, P > 0.05); the ascites parasite loads were (15.00 ± 5.00) × 10⁴, (6.67 ± 2.89) × 10⁴, (6.67 ± 2.89) × 10⁴, (8.33 ± 2.89) × 10⁴ and (1 766.67 ± 2.33) × 10⁴ parasites per mL respectively, with statistically significant difference [F(4,10) = 123 320, P < 0.01]. The qPCR results showed that on day 7 post infection, the relative Toxoplasma parasite loads in the heart, liver, spleen, lung, kidney and brain of mice in the MD-BMZ group were 2.32 ± 0.85, 2.32 ± 0.76, 1.00 ± 0.45, 1.00 ± 0.52, 4.46 ± 0.98 and 1.59 ± 0.63 respectively, all significantly lower than 7.28 ± 1.25, 7.11 ± 1.18, 3.46 ± 0.95, 7.28 ± 1.22, 9.37 ± 1.36 and 8.67 ± 1.29 in the NC group (t = 6.56, 6.83, 4.68, 9.47, 5.86, 9.86, all P < 0.01). HE staining showed that BMZ treatment significantly reduced the inflammatory cell infiltration and granulomatous lesions in the spleen, kidney and liver, without causing obvious drug-related damage. Conclusion BMZ exhibited excellent anti-Toxoplasma activity both in vivo and in vitro, with superior efficacy to SD-Na and high safety at the medium dose.

Objective To investigate the species of freshwater snails and the prevalence of trematode infections in freshwater snails in Lahsa City, and to analyze the phylogeny of freshwater snails and trematodes. Methods Freshwater snail samples were collected from Chengguan District, Duilongdeqing District, Linzhou County, Dazi District and Qushui County of Lhasa City from June to July 2024, and characterized for species, and the morphology and infection of trematodes were observed using microscopy. Genomic DNA was extracted from freshwater snails and trematodes for amplification of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene of freshwater snails and the internal transcribed spacer 2 (ITS2) of trematodes using PCR assay, and the PCR amplification products were sequenced. Raw reads were assembled using the DNASTAR software, and the obtained target sequences were submitted to the NCBI database for sequence alignment using the BLAST tool. Phylogenetic trees were constructed using the neighbor-joining and maximum likelihood methods, and genetic distances were calculated using the software MEGA 11.0. Results A total of 1 022 freshwater snails were collected and 8 genera were morphologically identified. Multiple developmental stages of trematode larvae were observed using microscopy, including miracidia, sporocysts, rediae, cercariae, and metacercariae. PCR amplification displayed the CO1 fragment from freshwater snails with approximately 740 bp in length and the ITS2 fragment from trematodes with approximately 630 bp in length, which were consistent with the expected size of target fragments. Gene sequencing and sequence alignment revealed that the CO1 sequence of freshwater snails exhibited the highest sequence identity with those of Physella acuta (98.66%), Ampullaceana lagoti (99.23%), Bellamya aeruginosa (99.29%), Galba pervia (100%), Gyraulus sp. (98.78%), Orientogalba viridis (98.91%), Radix Auricularia (99.38%), R. makhrovi (100%) and Tibetoradix khamensis (99.65%), and phylogenetic analysis revealed 8 clusters, with species of each genus clustering independently into a single clade. The ITS2 sequence of trematodes exhibited 100% sequence identity with Diplostomum mergi, Cotylurus syrius, Apatemon sp., Plagiorchis maculosus, Fasciola hepatica, F. gigantica, and Paramphistomum cervi, and showed the highest sequence identity with Schistosoma turkestanicum (99.80%), Echinostoma revolutum (99.84%), and Notocotylidae sp. (98.78%), respectively. Phylogenetic analysis revealed 9 clusters, with species of each genus clustering independently into a single clade, and all clades were supported by high bootstrap values, generating stable monophyletic clades. The overall prevalence of trematode infects was 59.59% (609/1 022) in freshwater snails across five counties (districts), with the highest prevalence in Qushui County (73.80%, 138/187), and the lowest in Chengguan District (39.24%, 31/79). Dazi District exhibited the greatest species diversity of freshwater snails (7 species), whereas Duilongdeqing District and Linzhou County harbored the highest trematode species (9 species each). In addition, Qushui County had the fewest species of both freshwater snails (1 species) and trematodes (2 species). Conclusion Freshwater snail species are abundant in the high-altitude waters of Lhasa City, with diverse trematode species and high prevalence of trematode infections.

Objective To establish a rapid, convenient, and reliable immunological test for field screening and early detection of intestinal parasitic diarrhea in livestock, so as to address the practical needs of grassroots disease diagnosis and control. Methods The specific antigen genes of Cryptosporidium parvum cgd2-420, Giardia lamblia α-15 giardin, and Eimeria bovis heat shock protein 90 (hsp90) were amplified and the recombinant plasmids were constructed. The recombinant antigen proteins were expressed in Escherichia coli and subsequently purified. BALB/c mice were immunized with purified proteins to generate monoclonal antibodies, which were subsequently conjugated with colloidal gold nanoparticles. Specific monoclonal antibodies and goat anti-mouse IgG antibody were striped onto nitrocellulose membranes as test lines and control lines to generate a colloidal gold immunochromatographic test strip. The diagnostic performance of the test strip was systematically evaluated following optimization of colloidal gold labeling conditions, antibody labeling amounts, and coating concentrations of test and control lines, including sensitivity, specificity, repeatability, and stability. Results Recombinant C. parvum cgd2-420, G. lamblia α-15 giardin, and E. bovis HSP90 proteins were successfully constructed and expressed, with relative molecular masses of approximately 40 000, 33 000, and 50 000, respectively, which were consistent with their theoretical molecular masses. All these recombinant proteins exhibited good immunogenicity. Following immunization of BALB/c mice, and cell fusion, and screening, four, five, and five specific monoclonal antibodies were obtained targeting the three pathogens, respectively, and monoclonal antibodyclones HP2, CG5, and JD5 with optimal performance were screened for preparation of the test strip. Following optimization of the reaction condition, the best stability of the colloidal gold labeling system was achieved with addition of 5 μL of 0.2 mol/L K₂CO₃, and the optimal antibody labeling amount was 20 μg, while the optimal coating concentrations for the test and control lines were 2 mg/mL and 1.5 mg/mL, respectively. The immunochromatographic test strip exhibited a minimum detectable dilution of 1∶64, and showed no cross-reactivity with non-target pathogens, which produced consistent intra-batch and inter-batch detection results and maintained stable detection performance after storage under dry conditions at 4 ℃ for 60 days. This test strip showed high consistency with PCR assay for detection of 80 clinical samples, with coincidence rates of 93.75% for E. bovis, 96.25% for C. parvum, and 96.25% for G. lamblia, respectivelys. Conclusion A simple, rapid, and stable colloidal gold-based immunochromatographic assay has been successfully established for rapid detection of intestinal protozoa in livestock, which is suitable for field screening and early diagnosis of C. parvum, G. lamblia, and E. bovis. This assay provides a powerful technical support for early identification and control of parasitic diarrhea in livestock.

Objective As the national schistosomiasis control program enters a new phase in China and the core content of parasitology curricula remains overall stable, this study aimed to explore the specific pathway for systematically integrating the One Health concept into the current human-animal-environment teaching framework, investigate the evidence for cognitive construction and competency development and develop an operable instructional design model. Methods Based on the current medical parasitology courses and non-adjustment of essential schistosomiasis knowledge points and allocated class hours, the knowledge structure and classroom activity chain supporting active construction and situated learning were constructed through reshaping human-animal-environment-related contents across three modules (life cycle, pathogenesis, and public health control) based on literature analysis and summarizing current status of medical parasitology teaching, under the guidance of constructivist meaning-making, situated learning, and constructive alignment, and a teaching procedure was developed that included scenario-based introduction, case analysis, group scenario simulation and diversified assessment. The outcome measure was the percentage of the score for schistosomiasis items in the total score for parasitology items in the end-of-term pathogenic biology examination between the experimental and control classes during each session. Homogeneity of variance was tested before between-group comparisons, and difference of means between groups was compared with two-sided Welch’s two-sample t tests. Results A “source of infection-aquatic environment-snail habitats-human/animal exposure-disease burden” multi-level representational framework was constructed, which was centered on the life cycle of Schistosoma and coincided with the distribution of cognitive load. Based on retainment of core knowledge points of the textbook, this framework explicitly embedded human populations, animal hosts, and environmental factors to provide an external scaffold for diagram integration, so as to generate an integrated human populations-animal hosts-ecological environment three-domain instructional structure. In addition, an “eco-pathogenesis” analytic perspective was introduced into the pathogenesis module as a teaching tool to link pathophysiology and eco-epidemiology. Over four consecutive sessions, results from objective assessments showed no significant difference in the proportion of the score for schistosomiasis items in the total score for schistosomiasis items in the end-of-term pathogenic biology examination between the experimental and control classes in 2021 (73.80% vs. 70.32%; t = 0.800, P > 0.05), and higher percentages in experimental classes than in control classes from 2022 to 2024 [2022: 82.04% vs. 75.78%, t = 2.059, P < 0.05; 2023: 79.46% vs. 70.15%, t = 2.604, P < 0.05; 2024: 73.06% vs. 65.16%, t = 2.645, P < 0.05], with 6% to 9% superiority over the control class. Conclusion Without altering the basic teaching contents of schistosomiasis, structural refinement and contextual reinforcement of the current human-animal-environment framework based on constructivism and situated learning may provide a feasible route to translating One Health into an operable framework for organizing teaching contents and learning activities, rather than a simple overlap perspective. Compared with conventional teaching methods, this model delivers markedly superior teaching outcomes and offers a viable instructional approach to strengthening systematic comprehension and integrated control thinking.

To investigate the frequency of the I1532T mutation in the voltage-gated sodium channel (vgsc) of Aedes albopictus in Guangyuan City, Sichuan Province, adult Ae. albopictus mosquitoes were collected using CO₂-baited light trapsfrom residential areas in seven counties (districts) of Guangyuan City between June and July in 2024. Genomic DNA was extracted from mosquitoes, and the genotype at the vgsc 1532 locus was detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. The detection performance of PCR-RFLP assay was validated by sequencing. Hardy-Weinberg equilibrium (HWE) test for populations was assessed using the software Genepop 4.7, and Fisher’s exact test was used to compare allele frequencies across counties (districts) using the software SPSS 25. A total of 208 Ae. albopictus mosquitoes were detected. PCR amplification of all samples showed bands with approximately 220 bp in length, which was consistent with the expected size. The frequencies of the vgsc wild-type homozygous 1532I/I genotype, wild-type/mutant heterozygous 1532I/T genotype, and resistant homozygous 1532T/T genotype were 87.98% (183/208), 10.58% (22/208), and 1.44% (3/208) in Guangyuan City, respectively. Among 7 counties (districts), the frequencies of wild-type/mutant heterozygous 1532I/T genotype were 12.5% (4/32), 9.38% (3/32), 17.86% (5/28), 28.13% (9/32) and 1/16 in Jiange County, Lizhou District, Qingchuan County, Wangcang County and Zhaohua District, respectively, while no heterozygous genotype were found in Chaotian District or Cangxi County. The frequencies of resistant homozygous 1532T/T genotype were 6.25% (2/32) and 3.13% (1/32) in Lizhou District and Wangcang County, respectively, and this homozygous genotype were not detected in other counties (districts). The frequencies of the vgsc wild-type 1532I and mutant 1532T alleles were 93.27% (388/416) and 6.73% (28/416) in Guangyuan City, respectively, and the frequencies of mutant 1532T alleles were 0 (0/64), 0 (0/72), 6.25% (4/64), 10.94% (7/64), 8.93% (5/56), 17.19% (11/64), and 3.13% (1/32) in Chaotian District, Cangxi County, Jiange County, Lizhou District, Qingchuan County, Wangcang County, and Zhaohua District, respectively (P < 0.05). HWE test showed that the Ae. albopictus population in Lizhou District deviated significantly from equilibrium (P < 0.05), and Ae. albopictus populations from Jiange County, Qingchuan County, and Wangcang County were at equilibrium (P > 0.05), while Ae. albopictus populations from other three counties (districts) were not suitable for HWE tests due to absence of the 1532T mutation (Chaotian District and Cangxi County) or low mutation frequency (Zhaohua District). The verification results showed that the coincidence rate between PCR-RFLP and sanger sequencing results was 100%. This study indicates a relatively high frequency of the vgsc I1532T mutation in Ae. albopictus populations from Guangyuan City, with resistant homozygotes detected in Lizhou District and Wangcang County.

To clarify the infection source and genetic characteristics of Giardia lamblia isolated from a diarrheal patient, fresh fecal sample of the patient was collected. Microscopy, real-time fluorescent quantitative PCR and nested PCR were comprehensively applied for pathogen detection. Metagenomic sequencing and analysis were performed targeting the small subunit ribosomal RNA (SSU rRNA), triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), β-giardin (bg) and translation elongation factor 1α (ef1α). Multilocus genotyping and genetic evolution analysis were carried out via sequence alignment and phylogenetic tree construction. Both microscopy and real-time fluorescent quantitative PCR confirmed Giardia infection. Only the gdh and ef1α genes were successfully amplified by nested PCR, and the SSU rRNA gene sequence was successfully detected by metagenomic sequencing. Multilocus genotyping showed that the isolate DXH001 obtained in this study belonged to assemblage B, and no mixed infection was detected. Phylogenetic analysis indicated that the gdh gene of isolate DXH001 shared 100% sequence identity with the human-derived strain EU847734 from Belgium, belonging to subtype BⅣ; the ef1α gene shared 98.43% sequence identity with the human-derived isolate HQ179598 from Australia, and the SSU rRNA gene shared 90.45% sequence identity with the human-derived strain U09491 from the United States, both clustered into the assemblage B branch. Compared with the human-derived reference sequence U09491, the SSU rRNA gene had 35 base substitutions and 5 deletions, and the sequence difference was mainly related to the insufficient sequencing data volume caused by the low abundance of the target sequence in the sample; the variations of gdh and ef1α genes were relatively conservative, which did not cause significant functional changes. In this study, the isolate DXH001 was identified as G. lamblia assemblage BⅣ via multilocus genetic analysis, suggesting the trans-regional transmission risk of this subtype. Metagenomic sequencing shows high sensitivity in low-abundance target detection, which can serve as an effective supplement for conventional molecular genotyping.

Whole blood samples were collected from international students studied in China for the first time from malaria-hyperendemic regions who underwent health examinations at the Hangzhou International Travel Healthcare Center in summer and autumn during the period from 2024 to 2025, and were tested for Plasmodium using a Plasmodium antigen colloidal gold assay, blood smear microscopy, and fluorescent quantitative PCR assay (qPCR). Basic demographics and epidemiological history were collected from students tested positive for Plasmodium. A total of 530 international students from 20 countries were tested, and Plasmodium antigen colloidal gold assay tested 22 positives, with a positive rate of 4.2% (22/530), including 16 positives from Nigeria, 2 from Tanzania, 2 from Guinea, one from Democratic Republic of Congo, and one from the Republic of Congo. Among the 22 positive cases, 11 cases were positive for P. falciparum detected by qPCR, and 1 case was positive by blood smear microscopy. Epidemiological surveys revealed that all positive individuals were afebrile and asymptomatic during healthy examinations. One microscopically positive case was hospitalized for treatment, and one case was hospitalized for treatment due to fever following healthy examinations. These data indicate asymptomatic cases among international students studied in China from malaria-endemic regions. Enhanced malaria surveillance and health education is required among international students studied in China to facilitate early detection and treatment, so as to reduce the risk of overseas importation of malaria.

A 70-year-old female, herdsman from Gonghe County, Qinghai Province, was presented to Department of General Surgery, Qinghai Provincial People’s Hospital on November 8, 2025, with complaints of lower abdominal pain and discomfort for 2 years and aggravation for 4 days. The patient has a clear history of long-term residence in a pastoral area. Abdominal and pelvic CT scans displayed a massive cystic mass in the abdomen and pelvis, containing multiple daughter cysts and band-shaped high-density shadows, with the largest cross-sectional dimension measuring approximately 225 mm × 85 mm, compression of the right ureter and hydronephrosis, gallstones and a cyst in the left kidney. Color doppler ultrasound of the pelvis showed a cystic mass measuring approximately 225 mm × 85 mm from the xiphoid process to the pelvic cavity, and the mass displayed clear borders and contained multiple daughter cysts with a “cabbage-like” appearance. Admission diagnosis included intraabdominal echinococcosis, urinary tract infection and chronic hepatitis B. The case was given exploratory laparotomy, complete excision of Echinococcus cysts and adhesiolysis of pelvic adhesions under general anesthesia on November 13, 2025, based on imaging findings and clinical symptoms. Intraoperative findings included a massive cystic mass in the abdominal and pelvic cavities, measuring approximately 22 cm × 8 cm, with thick, milky-white walls. Exploration revealed that the pedicle of the mass originated from the right ovary, and the main body of the cyst extended upward into the upper abdomen and was firmly adherent to the greater omentum, the inferior margin of the liver, the right kidney, and the right ureter, with the uterus and left adnexa appearing normal. Right ovarian echinococcosis was diagnosed intraoperatively, and cystectomy of right ovarian echinococcal cysts was performed. Following postoperative pathological examinations, right ovarian echinococcosis was confirmed. The patient recovered well postoperatively and was discharged from hospital 9 days post-operation. The patient was instructed to take albendazole (200 mg/dose, twice daily) regularly after discharge and to undergo periodical follow-up with color Doppler ultrasound of the liver, gallbladder, pancreas, and spleen, as well as liver function tests. No recurrence or surgery-related complications have been recorded until now.