Table of Content

30 October 2025, Volume 43 Issue 5

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EDITORIAL

Parasitic disease control for seventy-five years, embarking on a new journey to safeguard health: The 75th anniversary of the National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (National Center for Tropical Diseases Research)

LI Shizhu, ZHOU Xiaonong

2025, 43(5):  603-606.  doi:10.12140/j.issn.1000-7423.2025.05.001

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In 1950, the East China Branch of the Central Academy of Health was established in Nanjing. After multiple relocations, expansions, reforms, and renaming, it has developed into the National Institute of Parasitic Diseases (NIPD), Chinese Center for Disease Control and Prevention (National Center for Tropical Diseases Research), which is located in Shanghai today. NIPD has made great contributions to the research and control of schistosomiasis, malaria, filariasis, visceral leishmaniasis (kala-azar) and hookworm disease in China and globally during the past 75 years, and these endeavors have yielded remarkable achievements. Today, NIPD has become a leading technical center for guiding the control, research, teaching, and international collaboration of parasitic and tropical diseases in China, and plays a pivotal role in safeguarding public health, advancing the construction of “Healthy China”, and contributing to the global community with a shared future for humanity. This article provides a brief introduction to the 75-year development of NIPD.

CONSENSUS AND GUIDELINE

Expert consensus on species identification of Trichinella spp. by nucleic acid method (2025)

ZHANG Xi, CUI Jing, LIU Ruodan, LONG Shaorong, CHEN Muxin, CHEN Jiaxu, FU Baoquan, LIU Xiaolei, LIU Mingyuan, ZHANG Yalan, ZHANG Hongwei, WANG Zhongquan

2025, 43(5):  607-613.  doi:10.12140/j.issn.1000-7423.2025.05.002

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The larvae of Trichinella spp. nematodes parasitize in humans and various vertebrates, which seriously endangers people’s health and animal meat food safety. There are numerous species within the Trichinella genus; and the infectivity and pathogenicity of Trichinella spp. larvae are obviously different. In recent years, new Trichinella species have been discovered consecutively. The available pathogenic examination methods(direct microscopy of biopsy muscles and artificial digestion) could not distinguish the Trichinella species. To provide reference for Trichinella taxonomic researches and the accurate diagnosis of trichinellosis, this consensus elaborate on the nucleic acid methods for the identification of Trichinella spp. based on the latest domestic and international research progress.

ORIGINAL ARTICLES

Epidemiological characteristics of echinococcosis in Xinjiang Production and Construction Corps from 2014 to 2023

YU Yale, GONG Panlong, ZHAO Qian, CHENG Yanbiao, TIAN Siyang, WANG Tongmin

2025, 43(5):  614-618.  doi:10.12140/j.issn.1000-7423.2025.05.003

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Objective To understand the epidemiological characteristics and trends of echinococcosis in Xinjiang Production and Construction Corps from 2014 to 2023, and to provide insights for further prevention and control of echinococcosis. Methods Echinococcosis cases reported in Xinjiang Production and Construction Corps from January 1, 2014, to December 31, 2023, were collected from the National Infectious Disease Reporting Information Management System. Descriptive statistical analysis of the data was performed with SPSS 26.0. Chi-square test was used for comparison of rates. Results A total of 697 cases were reported in 13 divisions of Xinjiang Production and Construction Corps from 2014 to 2023, with no deaths, and the annual average incidence was 2.32/10⁵. The incidence showed an overall trend of first increasing and then decreasing over the years, peaking in 2017. The reported cases were mainly distributed in the 4th division (186 cases), 8th division (101 cases), and 9th division (67 cases) in Northern Xinjiang, and the 3rd division (56 cases) in Southern Xinjiang, accounting for 58.8% (410/697) of the total reported cases. The reported incidence in the Northern Xinjiang (2.77/10⁵) was higher than that in the southern Xinjiang (1.21/10⁵) (χ² = 66.353, P < 0.01). There were 368 male and 329 female reported cases. The average annual incidence was 2.34/10⁵ for males and 2.29/10⁵ for females, and the difference was not statistically significant (χ² = 0.097, P > 0.05). The mean age of the reported cases was 42.6 years (range 2 to 97 years), with the highest proportion at ages of 40-< 50 years (23.2%, 162/697), followed by 50-< 60 years (22.2%, 155/697), and the incidence among different age groups was statistically significant (χ² = 168.952, P < 0.001). The largest occupational group was farmers (31.1%, 217/697), followed by retirees (15.1%, 105/697). Conclusion The reported prevalence of echinococcosis in Xinjiang Production and Construction Corps showed an overall downward trend from 2014 to 2023; however, a risk of transmission persists.

Effectiveness of an auxiliary diagnosis system for ultrasound classification of cystic echinococcosis based on the convolution neural network: A multicenter study

TIAN Jing, WANG Xiaorong, YILIYASI Abuduaini, YANG Lingfei, CHEN Lu, GAO Hongying, ZHOU Na, LI Yuling, CUI Liang, SONG Tao

2025, 43(5):  619-626.  doi:10.12140/j.issn.1000-7423.2025.05.004

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Objective To examine the value of the convolution neural network-based ResNet50 model in assisting ultrasound classification of hepatic cystic echinococcosis (HCE). Methods Ultrasound images of intrahepatic lesions were retrospectively collected from HCE patients that were definitely diagnosed with postoperative pathology or long-term follow-up of multiple imaging tools admitted to five medical institutions including First Affiliated Hospital of Xinjiang Medical University (groups A to E, among them, group A is a provincial-level tertiary grade A hospital, groups B to D are prefecture level tertiary grade A hospitals, and group E is a county-level secondary Grade A hospital; and four grassroots hospitals were merged into group F and 5 hospitals merged into group G) during the period from January 2020 through June 2024. All images were classified by a junior sonographer and a senior sonographer according to five types of CE1 (univesicular cysts), CE2 (multiple daughter cysts), CE3 (cyst with detachment of laminated membrane), CE4 (heterogeneous or hyperechoic degenerative contents), and CE5 (calcified cysts), and uploaded to the ResNet50 model-based intelligent classification diagnosis ultrasound imaging cloud platform for classification diagnosis of HCE. The diagnostic accuracy of HCE classification by junior sonographers, senior sonographers and ResNet50 model was compared among groups using chi-square test, and pairwise comparisons were conducted with Bonferroni correction. In addition, the diagnostic consistency was evaluated with kappa analysis. Results A total of 1 596 ultrasound images of HCE were included from 952 HCE patients. The diagnostic accuracy of HCE classification by junior sonographers, senior sonographers and ResNet50 model were 88.3% (594/673), 95.7% (644/673) and 89.2% (600/673) in Group A, 72.7% (157/216), 86.6% (187/216), and 89.8% (194/216) in Group B, 77.6% (232/299), 87.6% (262/299) and 91.3% (273/299) in Group C, 74.5% (164/220), 85.5% (188/220) and 90.5% (199/220) in Group D, 69.1% (130/188), 84.0% (158/188) and 88.3% (166/188) in Group E, 75.1% (693/923), 86.1% (795/923) and 90.1% (832/923) in Group F and 80.6% (1 287/1 596), 90.2% (1 439/1 596) and 89.7% (1 432/1 596) in Group G. The overall diagnostic accuracy of HCE classification was lower by the ResNet50 model diagnosis than by senior sonographers in Group A (χ² = 27.140, P < 0.05), and the overall diagnostic accuracy was higher by the ResNet50 model than by junior sonographers in groups B, C, D, and E (χ² = 25.381, 24.307, 21.121 and 24.213; all P < 0.05). The overall diagnostic accuracy of HCE classification was higher by the ResNet50 model than by junior and senior sonographers in Group F (χ² = 82.654, P < 0.05), and the overall diagnostic accuracy was higher by the ResNet50 model than by junior sonographers in Group G (χ² = 80.749, P < 0.05). Moreover, all pairwise comparisons after Bonferroni correction were statistically significant with P < 0.025. The ResNet50 model demonstrated the highest diagnostic consistency with senior sonographers for HCE classification across all groups (kappa ≥ 0.836), followed by between junior and senior sonographers (0.754 ≤ kappa ≤ 0.863), and the lowest diagnostic consistency was seen between the ResNet50 model and junior sonographers (0.674 ≤ kappa ≤ 0.855). In the ResNet50 model, CE1 was most likely to be misdiagnosed as CE3 (68.6%, 24/35), CE2 misdiagnosed as CE4 (37.9%, 11/29) or CE3 (31.0%, 9/29), CE3 misdiagnosed as CE2 (65.6%, 21/32), and CE4 and CE5 were likely to be mutually misdiagnosed (61.9%, 26/42; 57.9%, 22/38). Conclusion The convolutional neural network-based ResNet50 model is effective to decipher the ultrasound features of HCE classification, which provides auxiliary diagnostic supports to doctors in primary medical institutions, and improves the accuracy of classification interpretation.

Echinococcus granulosus native antigen B regulating macrophage polarization through the STAT6/PPAR-γ signaling pathway in vitro

CAO Li, YANG Xuehua, JIAO Hongjie, HE Baiqifeng, ZHANG Yunfei, YUE Yingbin, CHENG Yongfeng, WANG Jiajing, SONG Haichen, YAN Mei

2025, 43(5):  627-634.  doi:10.12140/j.issn.1000-7423.2025.05.005

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Objective To investigate the regulatory effect of native antigen B (nAgB) from Echinococcus granulosus on the polarization of RAW264.7 macrophages in vitro and its potential involvement in the Signal Transducer and Activator of Transcription 6 (STAT6)/peroxisome proliferator-activated receptor-γ (PPAR-γ) signaling pathway. Methods nAgB was extracted and purified under sterile conditions. RAW264.7 cells were treated with nAgB at concentrations of 0, 1, 4, 16, and 64 μg/ml. Cell viability was assessed at 0, 6, 12, 24, and 48 hours using the CCK8 assay, and cell proliferation was evaluated at 24 hours via 5-ethynyl-2-deoxyuridine (EdU) staining. Cells were divided into 6 groups: control group, M1-polarized group, M2-polarized group, nAgB group, nAgB-treated M1-polarized group, and nAgB-treated M2-polarized group. The nAgB, nAgB-treated M1-polarized, and nAgB-treated M2-polarized groups were treated with 1 μg/ml nAgB. After 1 hour of intervention, the M1-polarized and nAgB-treated M1-polarized groups were stimulated with 500 ng/ml lipopolysaccharide (LPS) and 100 ng/ml interferon-γ (IFN-γ), while the M2-polarized and nAgB-treated M2-polarized groups were stimulated with 100 ng/ml interleukin-4 (IL-4) and 100 ng/ml IL-13. Cells and culture supernatants were collected from all groups. The relative mRNA transcription levels of macrophage polarization markers, STAT6, and PPAR-γ were detected by qPCR. The relative protein expression levels of relevant markers were measured by Western blotting, and the concentrations of related cytokines in the supernatant were determined by ELISA. Results CCK8 assay results at 24 hours showed that cell viability in the 1, 4, 16, and 64 μg/ml nAgB-treated groups (1.41 ± 0.09, 1.62 ± 0.08, 1.78 ± 0.04, 1.90 ± 0.04, respectively) was significantly higher than in the 0 μg/ml group (1.07 ± 0.05) (t = 8.67, 14.57, 27.43, 31.87; all P < 0.01). EdU assay results indicated that the cell proliferation capacity in the 1, 4, 16, and 64 μg/ml nAgB-treated groups (0.65 ± 0.01, 0.78 ± 0.02, 0.89 ± 0.02, 0.99 ± 0.02, respectively) was significantly higher than in the 0 μg/ml group (0.47 ± 0.02) (t = 16.21, 20.58, 33.47, 39.43; all P < 0.01). qPCR results showed that the relative mRNA levels of inducible nitric oxide synthase (iNOS), IL-6, and IL-1β in the nAgB-treated M1-polarized group (3.23 ± 0.92, 1.56 ± 0.58, 19.87 ± 0.35) were significantly lower than those in the M1-polarized group (10.62 ± 1.68, 3.97 ± 0.25, 24.40 ± 0.03) (t = 6.69, 6.62, 22.07; all P < 0.01). Conversely, the relative mRNA level of Arg-1 in the nAgB-treated M2-polarized group (29.30 ± 2.92) was significantly higher than that in the M2-polarized group (14.94 ± 0.77) (t = 8.23, P < 0.01). The relative mRNA levels of STAT6 and PPAR-γ in the nAgB group (59.12 ± 3.03, 7.82 ± 0.50) were significantly higher than those in the control group (26.38 ± 1.89, 3.71 ± 0.17) (t = 15.90, 13.40; both P < 0.01), in the nAgB-treated M1-polarized group (40.73 ± 2.91, 4.19 ± 0.88) were higher than in the M1-polarized group (17.93 ± 1.90, 1.76 ± 0.08) (t = 11.37, 4.75; both P < 0.01), and in the nAgB-treated M2-polarized group (140.50 ± 5.64, 11.67 ± 0.80) were higher than in the M2-polarized group (37.55 ± 5.92, 6.87 ± 0.28) (t = 21.82, 9.84; both P < 0.01). Western blotting results showed that the relative protein expression of iNOS in the nAgB-treated M1-polarized group (0.60 ± 0.02) was significantly lower than in the M1-polarized group (1.02 ± 0.03) (t = 21.86, P < 0.01), while the expression of CD206 in the nAgB-treated M2-polarized group (1.03 ± 0.04) was significantly higher than in the M2-polarized group (0.84 ± 0.02) (t = 7.78, P < 0.01). The relative protein levels of p-STAT6 and PPAR-γ in the nAgB group (0.76 ± 0.03, 0.77 ± 0.02) were significantly higher than in the control group (0.55 ± 0.05, 0.37 ± 0.00) (t = 6.11, P < 0.05; t = 40.16, P < 0.01), in the nAgB-treated M1-polarized group (0.60 ± 0.01, 0.42 ± 0.04) were higher than in the M1-polarized group (0.39 ± 0.05, 0.18 ± 0.01) (t = 6.64, 10.06; both P < 0.01), and in the nAgB-treated M2-polarized group (1.12 ± 0.11, 0.94 ± 0.02) were higher than in the M2-polarized group (0.86 ± 0.05, 0.66 ± 0.00) (t = 3.71, 28.18; both P < 0.01). ELISA results revealed that the secretion of TGF-β1 and IL-10 in the nAgB group [(70.27 ± 4.57), (167.00 ± 29.27) pg/ml] was significantly higher than in the control group [(29.87 ± 2.24), (50.17 ± 8.99) pg/ml] (t = 13.76, 6.61; both P < 0.01). In the nAgB-treated M1-polarized group, the levels of TNF-α and IL-1β [(523.20 ± 6.72), (387.80 ± 3.84) pg/ml] were significantly lower, while the levels of TGF-β1 and IL-10 [(47.15 ± 0.98), (137.30 ± 9.80) pg/ml] were significantly higher, compared to the M1-polarized group, [(995.70 ± 9.92), (680.90 ± 3.33) pg/ml] and [(18.05 ± 0.57), (21.66 ± 0.07) pg/ml], respectively (t = 68.32, 99.90, 44.41, 20.44; all P < 0.01). Similarly, nAgB-treated M2-polarized group,the levels of TNF-α and IL-1β [(398.50 ± 2.57), (85.18 ± 5.14) pg/ml] were significantly lower, while the levels of TGF-β1 and IL-10 [(293.20 ± 15.09), (341.20 ± 77.94) pg/ml] were significantly higher, compared to the M2-polarized group [(578.70 ± 12.36), (157.60 ± 14.25) pg/ml] and [(167.20 ± 20.34), (72.44 ± 5.28) pg/ml], respectively (t = 24.71, 8.28, 8.62, 5.82; all P < 0.01). Conclusion nAgB promotes macrophage polarization towards the M2 phenotype and suppresses the M1 response. This immunomodulatory effect is potentially mediated through the activation of the STAT6/PPAR-γ signaling pathway.

Mechanism of neuroinflammation exacerbated by microglial pyroptosis induced by chronic Toxoplasma gondii infection

CHEN Kexu, SUN Yanxin, HONG Xinyu, REN Liqin, LI Xiaoran, PAN Wei, ZHANG Yumei

2025, 43(5):  635-642.  doi:10.12140/j.issn.1000-7423.2025.05.006

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Objective To unravel the mechanism of aggravation of neuroinflammation induced by microglial pyroptosis following chronic infection with the Toxoplasma gondii Chinese 1 genotype Wh6 strain (TgCtwh6). Methods Twenty C57BL/6 mice were divided into a control group and an infection group randomly, of 10 mice each group. Mice in the infection group were each administered by oral gavage with 10 TgCtwh6 strain cysts suspended in 200 μl PBS to establish a chronic T. gondii infection model, while mice in the control group received an equal volume of PBS via oral gavage. Mouse cerebral cortex was collected, and the differential expression profiles of pyroptosis-related genes were screened in the mouse cerebral cortex using transcriptome sequencing 6 weeks post-infection. Sixty C57BL/6 mice were divided into a control group, an infection group, a VX-765 control group, and a VX-765 treatment group randomly, of 15 mice each group. Mice in the infection group and VX-765 treatment group were each administered by oral gavage with 10 TgCtwh6 strain cysts. Mice in the VX-765 control group and VX-765 treatment groups were intraperitoneally injected with the caspase-1 inhibitor VX-765 (at a dose of 50 mg/kg) once every 2 days since 4 weeks post-infection for 7 injections, and mouse cerebral cortex was collected from each group 8 weeks post-infection. BV2 mouse microglial cells were assigned to a control group, an infection group, a VX-765 control group, and a VX-765 treatment group, of 5 × 10⁵ cells each group. Cells in the infection group and VX-765 treatment group were infected with an equal amount of TgCtwh6 strain tachyzoites, while cells in the VX-765 control and VX-765 treatment groups were treated with 20 μmol/L VX-765. Cells were harvested 24 h following culture. RNA was extracted from mouse cerebral cortex and BV2 cells, and the relative RNA expression of pyroptosis-related genes caspase-1 and gasdermin D (GSDMD) and pro-inflammatory cytokines interleukin-18 (IL-18), IL-1β, IL-6, tumor necrosis factor-α (TNF-α) were quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay. Protein was extracted from mouse cerebral cortex and BV2 cells, and the expression of caspase-1, caspase-1 p20 subunit (caspase-1 p20), GSDMD, and GSDMD N-terminal fragment (GSDMD-N) were determined using Western blotting assay. Comparisons of means between groups were done using independent sample t-test, and multi-group comparisons were conducted with one-way analysis of variance (ANOVA) and Tukey’s post hoc test. Results Transcriptome sequencing revealed upregulation of pyroptosis-related genes NOD-like receptor thermal protein domain associated protein 3 (NLRP3), caspase-1, caspase-4, GSDMD, IL-1β and IL-18 expression in the mouse cerebral cortex in the infection group, with relatively higher caspase-1 (16.48 ± 6.40 vs. 1.33 ± 0.42; t = 4.09, P < 0.05) and GSDMD (12.80 ± 5.62 vs. 0.59 ± 0.20; t = 3.76, P < 0.05) expression in the infection group than in the control group. RT-qPCR assay quantified relatively higher mRNA levels of caspase-1 (4.04 ± 0.38 vs. 0.88 ± 0.18), GSDMD (9.67 ± 0.27 vs. 1.00 ± 0.26), IL-18 (1.49 ± 0.16 vs. 0.97 ± 0.16), IL-1β (7.50 ± 0.27 vs. 0.94 ± 0.21), IL-6 (4.96 ± 0.79 vs. 0.92 ± 0.22) and TNF-α (9.97 ± 1.77 vs. 0.82 ± 0.42) in the mouse cerebral cortex in the infection group than in the control group [Tukey’s Honest Significant Difference test (Tukey’s HSD test), all P < 0.01], and lower relative mRNA expression of caspase-1 (1.13 ± 0.13), GSDMD (0.87 ± 0.25), IL-18 (0.77 ± 0.05), IL-1β (0.89 ± 0.11), IL-6 (1.03 ± 0.05), and TNF-α (0.93 ± 0.43) in the VX-765 treatment group than in the infection group (Tukey’s HSD test, all P < 0.05). Western blotting assay determined higher relative expression of GSDMD (1.49 ± 0.14 vs. 0.41 ± 0.29), caspase-1 (1.38 ± 0.24 vs. 0.50 ± 0.29), GSDMD-N (1.60 ± 0.17 vs. 0.70 ± 0.30), and caspase-1 p20 (0.89 ± 0.11 vs. 0.17 ± 0.06) in the mouse cerebral cortex in the infection group than in the control group (Tukey’s HSD test, all P < 0.05), and lower relative expression of GSDMD (0.76 ± 0.11), caspase-1 (0.43 ± 0.15), GSDMD-N (0.72 ± 0.29), and caspase-1 p20 (0.43 ± 0.14) in the VX-765 treatment group than in the infection group (Tukey’s HSD test, all P < 0.05). RT-qPCR assay detected relatively higher mRNA expression of caspase-1 (1.64 ± 0.03 vs. 0.94 ± 0.05), GSDMD (2.17 ± 0.40 vs. 0.81 ± 0.21), IL-18 (3.01 ± 0.31 vs. 1.02 ± 0.01), IL-1β (3.47 ± 0.05 vs. 0.99 ± 0.08), IL-6 (3.64 ± 0.15 vs. 0.94 ± 0.09), and TNF-α (2.23 ± 0.20 vs. 0.99 ± 0.03) in BV2 cells in the infection group than in the control group (Tukey’s HSD test, all P < 0.01), and significantly lower mRNA expression of caspase-1 (0.70 ± 0.06), GSDMD (1.29 ± 0.27), IL-18 (1.07 ± 0.01), IL-1β (0.98 ± 0.03), IL-6 (0.52 ± 0.03), and TNF-α (1.26 ± 0.03) in the VX-765 treatment group than in the infection group (Tukey’s HSD test, all P < 0.05). Western blotting assay determined higher relative expression of GSDMD (1.43 ± 0.34 vs. 0.67 ± 0.23), caspase-1 (1.45 ± 0.14 vs. 0.48 ± 0.02), GSDMD-N (1.29 ± 0.40 vs. 0.56 ± 0.22), and caspase-1 p20 (1.25 ± 0.11 vs. 0.41 ± 0.18) in BV2 cells in the infection group than in the control group (Tukey’s HSD test, all P < 0.05), and lower relative expression of GSDMD (0.78 ± 0.20), caspase-1 (0.61 ± 0.07), GSDMD-N (0.56 ± 0.22), and caspase-1 p20 (0.47 ± 0.17) in the VX-765 treatment group than in the infection group (Tukey’s HSD test, all P < 0.05). Conclusion Chronic infection with the TgCtwh6 strain induces microglial pyroptosis and promotes the release of pro-inflammatory cytokines, thereby exacerbating neuroinflammatory responses, while VX-765 inhibitor is effective to suppress pyroptosis and inflammatory responses.

Downregulation of IDO expression in decidual immune cells post-infection with Toxoplasma gondii result in adverse pregnancy outcomes during the first trimester of pregnancy

WANG Wenxiao, REN Liqin, ZHANG Han, YANG Ruohan, ZHANG Haixia, LIU Xianbing, HU Xuemei

2025, 43(5):  643-650.  doi:10.12140/j.issn.1000-7423.2025.05.007

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Objective To investigate the effect of Toxoplasma gondii infection on indoleamine 2,3-dioxygenase (IDO) expression in decidual immune cells at the maternal-fetal interface during pregnancy, and to examine its association with adverse pregnancy outcomes. Methods A total of 40 wild-type pregnant mice of the C57BL/6 strain were divided into the wild-type control group and the infection group, of 20 animals in each group, while 20 pregnant IDO gene-knockout mice were assigned as the IDO^-/- infection group. Mice in two infection groups were intraperitoneally injected with 250 T. gondii RH tachyzoites in 250 µl PBS on day 8 of gestation, and animals in the wild-type control group were intraperitoneally injected with an equal volume of PBS. The mental state of the pregnant mice was observed on day 14 of gestation, and placental weight of pregnant mice and fetal weight were recorded and the incidence of abnormal fetal mice was calculated. Clinical decidual tissues were obtained from healthy pregnant women who voluntarily chose to terminate their pregnancies during the first trimester, and mononuclear cell suspensions of human decidual tissues were prepared using the human lymphocyte separation solution. Cell suspensions were divided into the control group and T. gondii infection group, with 1.5 × 10⁷ cells in each group. No treatment was given in the control group, and cells in the T. gondii infection group were infected with T. gondii RH strain tachyzoites at a tachyzoite-to-cell ratio of 1 : 3. Cells were harvested in both groups 30 hours following incubation. The IDO expression was detected in decidual natural killer (dNK) cells, decidual macrophages (dMϕ), decidual dendritic cells (dDC), and decidual myeloid-derived suppressor cells (dMDSC) using flow cytometry. dNK cells, dMϕ, dDC, and dMDSC were separated and purified from human decidual mononuclear cells with CD3^-CD56⁺, CD14, DC, and CD33 magnetic microbeads separation kits, respectively. These types of cells were all divided into the control group and the T. gondii infection group, with 1 × 10⁷ cells in each group. Cells in the control group were given no treatment, while cells in the infection group were infected with T. gondii RH strain tachyzoites at a tachyzoite-to-cell ratio of 1 : 3. Cells in both groups were harvested 20 hours following culture. Total proteins were extracted from cells, and the IDO expression was determined in various types of decidual immune cells using Western blotting. All statistical analyses were performed using the software GraphPad Prism 10.1.2, and comparisons of means between groups were conducted with independent sample Student’s t-test. Results The pregnant mice exhibited poorer pregnancy outcomes, more listlessness, slower movements, more severe placental congestion, and placental dryness, collapse, and elasticity loss in the IIDO^-/- infection group relative to in the wild-type control group, and the pregnant mice in the IDO^-/- infection group had poorly developed fetuses and absorbed fetuses. The pregnant mice had lower placental [(55.73 ± 5.52) mg vs. (81.73 ± 9.35) mg; t = 6.329, P < 0.01] and fetal weights [(106.88 ± 37.33) mg vs. (198.66 ± 26.58) mg; t = 5.299, P < 0.01) in the wild-type infection group than in the wild-type control group, and the pregnant mice had lower placental [(39.87 ± 4.24) mg] and fetal weights [(51.67 ± 19.75) mg] in the IDO^-/- infection group than in the wild-type infection group (t = 5.726 and 3.241, both P values < 0.01). No abnormal fetal mice were seen in the wild-type infection group, while a higher incidence rate of abnormal fetuses was found in the wild-type infection group [(40.6 ± 20.69)%] than in the wild-type control group (t = 5.193, P < 0.01) and higher in the IDO^-/- infection group [(93.33 ± 10.80)%] than in the wild-type infection group (t = 5.600, P < 0.01). Flow cytometry detected no significant difference in the proportion of positive IDO expression in dNK cells between the infection and control groups [(8.02 ± 3.20)% vs. (9.65 ± 2.62)%; t = 2.025, P > 0.05], and higher proportions of positive IDO expression in dMϕ [(27.50 ± 6.24)% vs. (35.07 ± 8.11)%; t = 5.830, P < 0.05], dDC [(20.23 ± 3.20)% vs. (24.80 ± 4.13)%; t = 5.533, P < 0.05] and dMDSC [(3.14 ± 0.57)% vs. (6.81 ± 0.88)%; t = 5.644, P < 0.05] in the infection group than in the control group. The largest reduction in the proportion of positive IDO expression [(53.20 ± 11.56)%] was seen in dMDSC following T. gondii infection, followed by in dMϕ [(21.45 ± 3.65)%], dDC [(18.22 ± 4.17)%] and dNK cells [(17.84 ± 15.54)%]. Western blotting determined relatively lower IDO expression in dNK cells [(0.91 ± 0.10) vs. (1.04 ± 0.11); t = 13.360, P < 0.01], dMϕ [(0.62 ± 0.12) vs. (1.14 ± 0.09); t = 12.930, P < 0.01)], dDC [(0.75 ± 0.11) vs. (1.11 ± 0.06); t = 7.482, P < 0.05] and dMDSC [(0.47 ± 0.06) vs. (1.05 ± 0.03); t = 13.960, P < 0.01] in the infection group than in the control group. The largest reduction in the relative IDO expression [(55.22 ± 16.12)%] was seen in dMDSC following T. gondii infection, followed by in dMϕ [(46.30 ± 6.98)%], dDC [(33.53 ± 7.78)%] and dNK cells [(12.74 ± 0.67)%]. Conclusion T. gondii infection may downregulate the IDO expression in dNK cells, dMϕ, dDC and dMDSC at the maternal-fetal interface during the first trimester of pregnancy, thereby increasing the incidence of adverse pregnancy outcomes. These findings suggest that downregulation of IDO expression is closely associated with the occurrence of adverse pregnancy outcomes.

Repellent effect of nootkatone against Aedes albopictus

SU Yuanxin, CAI Jinghong, YIN Zige, LIU Qing, ZHENG Yaojie, GUO Ao, YAO Erming, ZHANG Huiwen, ZHAO Tongyan, ZHANG Hengduan, DENG Changsheng

2025, 43(5):  651-655.  doi:10.12140/j.issn.1000-7423.2025.05.008

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Objective To evaluate the repellent effect of the plant-derived repellent nootkatone against Aedes albopictus. Methods In the spatial repellency screening assay using three-part tubes, three replicates each of 20%, 10%, and 5% nootkatone solutions were tested, together with one blank control. For each group, 40 female Ae. albopictus mosquitoes (5 to 8 days post-emergence without blood feeding) were selected. The treatment side of the three-part tube was fitted with a mesh screen impregnated with the respective nootkatone solutions, while the control side was treated with absolute ethanol. Mosquitoes were introduced into the central chamber of the three-part tube, which were covered with green flannels. Then, the partitions to both sides were opened. After 10 minutes, the partitions were closed, and the number of mosquitoes in each side was recorded to calculate the relative repellency rate. The nootkatone solutions with a relatively higher repellency rate were selected for subsequent fabric efficacy and human hand topical application cage tests. For the fabric efficacy evaluation, five replicates were conducted, each utilizing 600 adult mosquitoes (3 to 5 days post-emergence without blood feeding, including 300 males and 300 females). Fabrics in the treatment group were immersed in the repellent at a dose of 7.5 ml/500 cm² for 48 hours, while the control group used untreated fabric from the same batch. The fabric was used to cover a 40 mm × 40 mm observation window on the back of test gloves. Five volunteers wore the gloves and inserted their hands into mosquito cages, and the number of mosquito probing attempts within 30 minutes was recorded. The criterion for complete protection was absence of any probing during this period. The residual efficacy was re-tested on days 5 and 7, and so forth, until protection failed. For the human hand topical application cage test, five replicates were performed, each using 300 female mosquitoes (3 to 5 days post-emergence without blood feeding). A 50 mm × 50 mm area on the back of a volunteer’s hand, aligning with the 40 mm × 40 mm observation window on the test glove, was marked. The volunteer wore a test glove exposing the treated area 2 hours post-treatment with nootkatone solution at a dose of 1.5 μl/cm², inserted their hands into the cage, and the number of probing attempts within 2 minutes was recorded. This test was repeated hourly, with the first occurrence of a probing event defined as the failure criterion, and the duration of effective protection was recorded. For the spatial repellency screening assay using three-part tube, Mann-Whitney U test was used to analyze the difference in mosquito distribution between the two sides under blank control conditions, and chi-square test was used to analyze differences between experimental groups (different concentrations) and the blank control. Results The spatial repellency screening assay using three-part tube showed a 15/15 mosquito distribution ratio between the control and treatment sides under blank conditions (Z = 0.145, P > 0.05). The relative repellency rates of 5%, 10%, and 20% nootkatone were 26.1%, 26.1%, and 57.7% against Ae. albopictus mosquitoes, respectively, with no statistically significant differences compared to the blank control (χ² = 3.112 1, 3.112 1, 0.331 4; P > 0.05). The repellency rate of 20% nootkatone was significantly different from that of both 5% and 10% nootkatone (χ² = 4.978, 4.978; P < 0.05). The fabric efficacy evaluation demonstrated that fabric treated with 20% nootkatone for 48 hours provided complete protection, with a protective duration of 5 days. The human hand topical application test indicated that the average effective protection time was 2.4 hours for 20% nootkatone. Conclusion In the fabric efficacy evaluation, 20% nootkatone provides a complete protection against Ae. albopictus.

Prevalence and phylogenetic characteristics of Anaplasma capra in parasitic ticks from selected regions of Fujian Province

HU Shiju, ZHOU Shuheng, WANG Jiaxiong, ZENG Zhiwei, HAN Tengwei, LIN Fen, WU Shenggen, XIAO Fangzhen

2025, 43(5):  656-662.  doi:10.12140/j.issn.1000-7423.2025.05.009

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Objective To investigate the prevalence and genetic evolutionary characteristics of Anaplasma capra in parasitic ticks in selected regions of Fujian Province. Methods Parasitic ticks were collected from domestic and wild animals (including murine, muntjacs, dogs, goats, hares, cattle, wild boars, and pigs) in Zherong County, Zhouning County, Fu’an City and Xiapu County of Ningde City and Pucheng County and Shaowu City of Nanping City from 2011 to 2024. Following morphological and molecular biological identification, genomic DNA was extracted from ticks and A. capra citratesynthase gene (gltA) gene was amplified using nested PCR assay. Positive PCR amplification products were sequenced and initially aligned by BLAST on National Center for Biotechnology Information (NCBI). Sequence alignment was performed using the software MEGA 11, and a phylogenetic tree was created using with the neighbor-joining method. Genetic diversity indices of A. capra gltA gene sequence were calculated using the software DnaSP 6.0, and a haplotype network was constructed using the software PopArt 1.7 after grouping sequences by region using the software Arlequin 3.5.2.2. Comparisons of proportions were done with chi-square test and Fisher’s exact test, and influencing factors were identified using a multivariate logistic regression model. Results A total of 650 ticks were tested, with the highest prevalence of A. capra infection on the muntjac body surface (19.11%, 30/157), followed by on cattle body surface 2/13, and there was a significant difference in the prevalence of A. capra infection in different hosts (χ² = 59.64, P < 0.01). The captured ticks were mainly adults, followed by nymphs. The highest prevalence of A. capra infection was detected in nymphs (15.38%, 8/52), followed by in adults (4.88%, 29/594), with no A. capra infection seen in larval ticks (0/2) or eggs (0/2), and there was no significant difference in the prevalence of A. capra infection in ticks at different developmental stages (χ² = 9.15, P < 0.05). The prevalence of A. capra infection was 7.99% (27/338) and 0.78% (2/256) in female and male adult ticks (χ² = 16.29, P < 0.01). The prevalence of A. capra infection was 5.86% (5/88) in ticks from Zherong County, 11.06% (23/208) in Zhouning County, 0/7 in Fu’an City and 0/7 in Xiapu County of Ningde City and 2.69% (9/335) in Pucheng County and 0/5 in Shaowu City of Nanping City, and there was no region-specific prevalence found (χ² = 15.74, P < 0.01). The prevalence of A. capra infection was 4.64% (7/151) in ticks sampled in summer, and 4.37% (9/206) in autumn, 7.34% (21/286) in winter, and no infection was detected in ticks sampled in spring (0/7) (χ² = 2.86, P > 0.05). Six hundred fifty ticks belonged to 14 species of 5 genera, and the overall prevalence of A. capra infection was 5.69% (37/650) in ticks. A. capra was detected in 7 tick species, with the highest prevalence in Ixodes ovatu (42.86%, 12/28), followed by in Haemaphysalis of unknown species in Fujian Province (31.58%, 12/38) (χ² = 83.16, P < 0.01). Multivariate analysis showed a significantly higher risk of A. capra infection in tick nymphs than in adult ticks (OR = 2.83, 95% CI: 1.15 to 6.94), and a lower risk of A. capra infection was seen in ticks from Nanping City than from Ningdu City (OR = 0.20, 95% CI: 0.08 to 0.50). BLAST analysis showed that the 37 sequences shared 97.88% to 98.92% identity with the A. capra reference sequence (GenBank accession number: OQ847085.1). Phylogenetic analysis revealed that the A. capra sequences from this study shared the closest relationship with sequences of A. capra from Qinghai Province. A total of 15 haplotypes were identified in sequences of A. capra from selected regions of Fujian Province, with overall haplotype diversity of 0.88 and nucleotide diversity of 0.009, and Hap1 to Hap3 were dominant haplotypes. Conclusion The prevalence of A. capra infection is high in parasitic ticks in selected regions of Fujian Province. Surveillance and management of tick-borne pathogens are recommended to be intensified.

Piroplasm infection and genetic diversity in Tibetan sheep in selected areas of Qinghai Province

HAN Yuan, LI Zhi, LIN Weishan, LI Chunhua, WANG Xiaohong, CAIRANG Zhouzai, LEI Mengtong

2025, 43(5):  663-669.  doi:10.12140/j.issn.1000-7423.2025.05.010

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Objective To investigate the prevalence of piroplasm infections and genetic evolutionary characteristics of piroplasm in Tibetan sheep from some areas in Qinghai Province. Methods Whole blood samples were collected from Tibetan sheep across 12 counties (districts) in 8 states (cities) of Qinghai Province, and genomic DNA was extracted using the silicon matrix column method. The 18S rRNA V4 hypervariable region sequences of piroplasm was amplified using nested PCR assay, and samples positive for nested PCR assay were sequenced, followed by sequence alignment using the BLAST program in NCBI to identify the piroplasm species. The sequences of samples positive for nested PCR assay were subjected to haplotype analysis and nucleotide polymorphism analysis using the software DnaSP version 6, and a haplotype network diagram was constructed using the PopArt software. A phylogenetic tree based on the 18S rRNA V4 hypervariable region sequence of piroplasm was built using the maximum likelihood method with the sequence of Babesia microti (GenBank accession number: AB242176) as the outgroup. Results A total of 475 Tibetan sheep blood samples were collected, and 193 blood samples were positive for piroplasm, with 40.6% (193/475) overall prevalence of infections. piroplasm infection was detected across 12 districts (counties), and the three highest prevalence seen in Menyuan (94.1%, 96/102), Haiyan (72.7%, 24/33), and Ledu (47.8%, 11/23). The species of piroplasm infecting Tibetan sheep included Theileria luwenshuni, T. ovis, T. uilenbergi and Theileria sp. OT3, with infection prevalence rates of 20.6% (98/475), 17.7% (84/475), 1.3% (6/475), and 1.1% (5/475), respectively. There were 13 haplotypes of T. luwenshuni, with TlH1 as the dominant haplotype (76.5%, 75/98), which were mainly distributed in Menyuan, Ledu, and Minhe, and there were 9 haplotypes of T. ovis, with ToH1 as the dominant haplotype (85.7%, 72/84), which were prevalent in Haiyan, Zeku, Menyuan, Gangcha, Huangyuan, Dari, Guinan, Tianjun, and Qilian, while there were 4 haplotypes of T. uilenbergi, with TuH2 as the dominant haplotype (3/6), which were mainly distributed in Menyuan, and there were 4 haplotypes of Theileria sp. OT3, with TsH3 as the dominant haplotype (2/5), which were mainly distributed in Minhe. Phylogenetic analysis revealed that the sequence of the T. luwenshuni 18S rRNA V4 hypervariable region from the whole blood of Tibetan sheep in Qinghai Province was clustered into a clade with the sequences of Budorcas taxicolor-derived T. luwenshuni isolate and sheep-derived T. luwenshuni isolates from Lanzhou, Tianzhu, Qinghai and Shaanxi in GenBank, and the sequence of Theileria sp. OT3 was clustered into a clade with those of Theileria sp. OT3 from yak-derived parasite isolates in Qinghai, goat-derived strains in Shaanxi and sheep-derived strains in Spanish. In addition, the sequence of T. ovis was clustered into a clade with published sequences of yak-derived strains from Qinghai, and sheep-derived strains from Qinghai, Sudan, Turkey, Egypt, and Xinjiang, while the sequence of T. uilenbergi was clustered into a clade with sequences of Qinghai vole-derived strains, goat-derived strains from Hunan and India, and sheep-derived strains from Iraq and Turkey. Conclusion The piroplasm infection in Tibetan Sheep were serious, with diverse species and haplotypes of the infected piroplasms in some areas from Qinghai Province, and the Theileria sp. OT3 was detected in Tibetan sheep, which provides a scientific support for the comprehensive prevention and control of piroplasmosis in Tibetan sheep in Qinghai.

Infection and genotype identification of Enterocytozoon bieneusi and Cryptosporidium in Wuzhishan pigs of Hainan

DONG Yatong, ZHANG Yan, LIU Guangliang, CAO Zongxi, YU Miao, CHEN Linlin, LIU Hailong, CHEN Suzhen

2025, 43(5):  670-677.  doi:10.12140/j.issn.1000-7423.2025.05.011

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Objective To analyze the species of Cryptosporidium and the genotype of Enterocytozoon bieneusi in Wuzhishan pigs in Hainan, and to master the infection of the two protozoa. Methods From June 2024 to February 2025, fresh fecal samples of Wuzhishan pigs were collected from 4 farms in Zuntan Town, Qiongzhong Li and Miao Autonomous County, Wuzhishan City and Chengmai County of Hainan Province (marked as A, B, C and D farms, respectively). DNA was extracted from fecal samples, and the 18S ribosomal RNA (18S rRNA) of Cryptosporidium and the internal transcribed spacer (ITS) sequence of E. bieneusi were amplified by nested PCR. The positive amplification products were sequenced and compared to identify Cryptosporidium species or E. bieneusi genotypes. SPSS 26.0 software was used for statistical analysis. Pearson chi-square (χ²) test was used to compare the differences in positive rates, and the risk coefficient (OR) and 95% confidence interval (CI) were calculated. The phylogenetic tree was constructed by MEGA 11 software adjacency method. Results A total of 137 Wuzhishan pig manure samples were collected, of which 41 were amplified with 830 bp of Cryptosporidium 18S rRNA sequence and 69 were amplified with 390 bp of E. bieneusi ITS sequence. The total positive rate of Cryptosporidium in Wuzhishan pigs from four farms was 29.9% (41/137), with the highest in C farm (64.3%, 18/28) and the lowest in B farm (20.3%, 14/69). The positive rate of Cryptosporidium in post-weaning piglets was 57.1% (12/21), which was higher than 7.7% (2/26) in pre-weaning piglets (χ² = 13.583, P < 0.05). The OR of post-weaning piglets and adult pigs was 16.00 times (95% CI = 2.98-85.99) and 5.14 times (95% CI = 1.15-23.31) higher than that of pre-weaning piglets. The total positive rate of E. bieneusi was 50.4% (69/137), with the highest in C farm (60.7%, 17/28) and the lowest in A farm (37.5%, 9/24). The positive rate of E. bieneusi in post-weaning piglets was 80.9% (17/21), which was higher than that in pre-weaning piglets (42.3%, 11/26) and adult pigs (45.5%, 41/90) (χ² = 7.204, 8.551; both P < 0.01). The OR of post-weaning piglets and adult pigs was 5.80 times (95% CI = 1.52-22.10) and 1.14 times (95% CI = 0.47-2.76) higher than that of pre-weaning piglets. The positive rate of E. bieneusi in grazing Wuzhishan pigs was 62.8% (44/70), which was higher than that in barn feeding Wuzhishan pigs (37.3%, 25/67) (χ² = 8.935, P < 0.01). The OR of infection in grazing Wuzhishan pigs was 2.84 times (95% CI = 1.42-5.69) higher than that in feeding Wuzhishan pigs. The results of sequence alignment showed that 41 Cryptosporidium 18S rRNA sequences were identified as C. scrofarum, and the sequence identity with C. scrofarum (GenBank accession number: PQ856498) was 93.36%-99.74%. In the phylogenetic tree, it was in the same large branch with C. scrofarum from Yunnan pig and Hubei pig (GenBank accession number: PQ856498, ON149807). 69 ITS sequences of E. bieneusi were identified as H, D, and EpbC genotypes. Among them, 3 H genotype sequences were 98.00% identical to the H genotype of German pig (GenBank accession number: AF135835.1). The identity of 25 D genotype sequences with D genotype (GenBank accession number: MK696083.1) from Australian cats was 97.51%-99.72%, and the identity of 41 EpbC genotype sequences with EpbC genotype (GenBank accession number: KY950535.1) from Chinese pandas was 98.59%-99.73%. In the phylogenetic tree, D genotype was in the same branch with Argentinean D genotype and Australian cat D genotype (GenBank accession number: OP650902, MK696083), EbpC genotype was in the same branch with Chinese panda EpbC genotype (GenBank accession number: KY950535.1), and H genotype was in the same branch with German pig H genotype (GenBank accession number: AF135835.1). Phylogenetic analysis showed that the three genotypes belonged to one subgroup. Conclusion There were E. bieneusi and Cryptosporidium infection in Wuzhishan pigs in Hainan. The Cryptosporidium infection species was C. scrofarum. The main genotypes of E. bieneusi were H, D and EbpC.

Laboratory detection of leishmaniasis in Henan Province from 2013 to 2023

LI Suhua, ZHAO Xiaofei, WANG Dan, JI Penghui, ZHOU Ruimin, ZHAO Yuling, YANG Chengyun, LIU Ying, ZHANG Hongwei, DENG Yan, HE Zhiquan

2025, 43(5):  678-682.  doi:10.12140/j.issn.1000-7423.2025.05.012

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Objective To analyze the laboratory detection results of leishmaniasis in Henan Province from 2013 to 2023, so as to provide data support for leishmaniasis control in Henan Province. Methods Suspected leishmaniasis cases’ anticoagulant whole blood samples, bone marrow smear samples and injured tissue samples were collected from the laboratory of Institute of Parasitic Disease Prevention and Control Institute Henan Center for Disease Control and Prevention from 2013 to 2023. All samples were collected from individuals retested positive for rK39 immunochromatographic test strips from all city-level disease control and prevention centers in Henan Province from 2013 to 2023, and cases’ characteristics were retrieved from the Infectious Diseases Surveillance System of the Information System of Chinese Center for Disease Control and Prevention. All samples were detected using microscopic examination, rK39 strips and PCR assay, and bidirectional sequencing was performed following PCR amplification. Successfully sequenced gene sequences were spliced and assembled using the software ChromasPro 1.5 and aligned using the BLAST tool. All statistical analyses were conducted using the statistical software SPSS 21.0. Differences of the positive rates of leishmaniasis by different assays were tested for statistical significance with Fisher’s exact test. Results A total of 186 samples were collected, originating from 137 suspected cases from 2013 to 2023, with the three highest numbers of cases from Anyang City (51, 37.23%), Zhengzhou City (44, 32.12%) and Luoyang City (31, 22.63%), and the highest number in 2023 (37.23%, 51/137). There were 94 cases from men and 43 from women, with a male to female ratio of 2.19:1, and leishmaniasis cases were predominantly at ages of 0-< 10 years (41, 29.93%) and 60-< 70 years (24, 17.52%). Microscopic examinations detected a positive rate of 62.00% in 50 bone marrow smears (31/50), and PCR assay and rK39 strips revealed positive rates of 66.92% (89/133) and 75.19% (100/133) in 133 anticoagulant whole blood samples, while PCR assay identified 2 positives in 3 injured tissue samples. A total of 92 cases were definitely diagnosed as leishmaniasis in 137 cases by means of microscopic examinations combined with PCR assay, with a positive rate of 67.15%. There was no significant difference in the positive rate of leishmaniasis between PCR assay and microscopic examinations (Fisher’s exact test, P > 0.05), and there were significant differences between microscopic examinations and rK39 strips and between PCR assay and rK39 strips (Fisher’s exact test, P < 0.05). The diagnostic sensitivity and specificity were 98.93% and 100% for PCR assay, 94.49% and 66.68% for rK39 strips and 76.92% and 90.91% for microscopic examinations, respectively. Seventy three out of 91 PCR assay-positive samples were successfully sequenced, and BLAST alignment revealed Leishmania infantum in 71 samples and L. major in 2 samples. Conclusion L. infantum remains predominant Leishmania species in Henan Province, with leishmaniasis cases primarily found from three cities of Anyang, Zhengzhou, and Luoyan. Intensified leishmaniasis control and improved laboratory detection capability are recommended.

Epidemiological characteristics of scrub typhus in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province from 2010 to 2024

GOU Changchun, LIU Aicong, FENG Kai, CAO Chunli, SONG Xiaoxiao, YUE Taike

2025, 43(5):  683-689.  doi:10.12140/j.issn.1000-7423.2025.05.013

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Objective To investigate the epidemiological characteristics of scrub typhus in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province from 2010 to 2024, so as to provide insights into optimization of scrub typhus control strategies. Methods Data on scrub typhus cases reported in Dehong from January 1, 2010, to December 31, 2024 were collected from the China Disease Prevention and Control Information System. The Joinpoint Regression Program version 5.3.0 was used to establish a Joinpoint regression model. The annual percent change (APC), average annual percent change (AAPC), and their 95% confidence intervals (CI) of the incidence of reported scrub typhus were calculated to analyze the trends in incidence by gender, region, and age group. Normality tests and descriptive statistical analyses were performed using the software SPSS 25.0. Results A total of 13 715 scrub typhus cases were reported in Dehong from 2010 to 2024, with no deaths, and the annual average reported incidence was 71.88/10⁵. The number of reported cases and reported incidence appeared an overall tendency towards a rise from 2010 to 2022, with an increase from 117 cases (9.84/10⁵) in 2010 to 1 817 cases (138.76/10⁵) in 2022. A slight decline was observed in 2023 (1 644 cases, 124.47/10⁵) and 2024 (1 457 cases, 108.97/10⁵), although the incidence remained high. There were significant differences in the number of reported cases and the incidence among years (χ² = 5 318.62, 4 783.42; both P < 0.01). Joinpoint regression analysis identified two turning points (in 2016 and 2022) and the trends in incidence were divided into three phases: a significant increase from 2010 to 2016 (APC = 33.61%, P < 0.01), a continued but slower increase from 2017 to 2022 (APC = 13.94%, P < 0.01), and a numerical decline from 2023 to 2024 (APC = -10.74%, P > 0.05). Mangshi, Yingjiang County and Longchuan County were high-incidence areas from 2010 to 2024, with 4 826 (35.19%), 3 921 (28.59%), and 3 151 (22.97%) reported cases, and average annual incidence rates of 77.57/10⁵, 84.83/10⁵, and 112.80/10⁵, respectively. Joinpoint regression analysis indicated that the reported incidence appeared a tendency towards a significant rise across all counties (cities) from 2010 to 2024 (all P < 0.01 for AAPC), with the largest increase seen in Lianghe County (AAPC = 48.00%), followed by Longchuan County (AAPC = 38.64%), Ruili City (AAPC = 22.57%), Mangshi (AAPC = 18.47%), and Yingjiang County (AAPC = 9.14%). A distinct seasonal pattern was observed in the incidence, with the peak in August (seasonal index = 2.65). The epidemic season was found from July to November, with the number of reported cases accounting for 84.81% (11 632/13 715) of the total reported cases. There were 7 799 (56.86%) male cases and 5 916 (43.14%) female cases, with a male-to-female ratio of 1.32 : 1, and the trends in incidence among both genders were consistent with the overall trend, with 2016 and 2022 as key turning points. Cases were widely distributed across all age groups, ranging from 1 month to 96 years, with a median age of 40 years. The majority of cases (55.49%, 7 611/13 715) were young and middle-aged adults aged 30 to 59 years. Joinpoint regression analysis revealed a significant increase in incidence across all age groups from 2010 to 2024 (all P < 0.01 for AAPC), with the largest increase seen among residents at ages of 40 to 49 years (AAPC = 19.41%, 95% CI: 17.36 to 24.60). Farmers were the dominant occupation (9 969 cases, 72.69%). Conclusion The incidence of scrub typhus appeared an overall tendency towards a rapid rise in Dehong from 2010 to 2024. Continuous improvements of disease surveillance and control is recommended tailored to high-risk regions, epidemic seasons, and high-risk populations.

Surveillance of foodborne parasite in market-sold food in Putuo District, Shanghai Municipality from 2017 to 2024

QIN Ruofang, XU Haiping, XU Yijiong, DAI Simin, LV Shan, CUI Yan

2025, 43(5):  690-696.  doi:10.12140/j.issn.1000-7423.2025.05.014

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Objective To investigate the contamination of foodborne parasites in market-sold and awareness of foodborne parasite knowledge and behaviors pertaining to foodborne parasites among residents in Putuo District, Shanghai Municipality from 2017 to 2024, so as to provide the evidence for formulating the control strategies for foodborne parasites in this district. Methods Freshwater fishes, freshwater shrimps, crabs and shellfishes, marine fishes, raw pickled aquatic products of animal origin, bullfrogs, and pasteurized milks were collected using a stratified random sampling method from different circulation links across different streets in Putuo District from 2017 to 2024, and the contamination of food-borne parasites was detected using the microscopy following dissection, artificial digestion method, and quantitative real-time PCR assay. Aquatic product sales personnel, service personnel and residents in food circulation places were randomly sampled using a stratified random sampling method from supermarkets, agricultural trade markets and restaurants from 2023 to 2024, and the awareness of foodborne parasitic diseases knowledge, habits of consumption of raw food and willingness to receive foodborne parasitic diseases control were investigated among participants using questionnaire surveys. Fisher’s exact probability test was used to compare proportions, followed by multiple comparisons with Bonferroni correction. Results A total of 1 055 food samples were collected and detected from 2017 to 2024, and 45 samples were tested positive for foodborne parasites, with an overall detection rate of 4.27%. The detection of Anisakis was 1.43% (33/105) in marine fish samples, with parasites predominantly detected in Scomber japonicus (3/3), Larimichthys polyactis (56.00%, 14/25), Trichiurus haumela (51.72%, 15/29) and Larimichthys crocea (1/17), and there was a significant difference in the prevalence of Anisakis infections among different types of marine fishes (Fisher’s exact probability test, P < 0.01). The detection of Echinostoma metacercariae was 11.32% (12/106) in raw pickled aquatic products of animal origin, and there was a significant difference in the detection of Echinostoma metacercariae among different types of raw pickled aquatic products of animal origin (Fisher’s exact probability test, P < 0.01). However, no foodborne parasites were detected in freshwater fishes, freshwater shrimps, crabs and shellfishes, bullfrogs, or pasteurized milk samples. The detection rates of Anisakis were 17.39% (4/23), 36.36% (28/77), 1/4, and 0/1 in marine fish samples from supermarkets and retail stores, agricultural trade markets, restaurants, and online stores, respectively (Fisher’s exact probability test, P > 0.05), and the detection rates of Echinostoma metacercariae were 18.18% (6/33), 17.14% (6/35), 0/13, and 0 (0/25) in raw pickled aquatic products of animal origin from supermarkets and retail stores, agricultural trade markets, restaurants, and online stores, respectively (Bonferroni correction, P > 0.05), indicating that the detection rate of foodborne parasites did not correlate sampling sites. Questionnaire survey showed that the awareness of foodborne parasitic diseases was 74.02% (94/127) among residents, with relatively higher awareness among participants at ages of 40 to 49 years (87.50%, 21/24), urban residents (76.07%, 89/117), participants with a bachelor degree (82.61%, 57/69), and healthcare professionals (100%, 32/32). Of all respondents, 61.42% (78/127) had behaviors of consumption of raw or semi-raw food, with a relatively higher proportion among participants at ages of 30 to 39 years (80.95%, 34/42) and postgraduates (7/10), and 96.06% (122/127) were willing to change their special dietary preferences or poor dietary habits. In addition, 97.64% (124/127) of respondents were willing to receive standardized treatment post-infection, and 53.54% (68/127) were willing to learn knowledge pertaining to foodborne parasitic diseases, with relative lower willingness among men (41.86%, 18/43), participants at ages of 18 to 29 years (44.74%, 17/38), suburban residents (4/10), retirees/unemployed persons (3/7), and aquatic food catchers/sellers (3/7). Conclusion There is a risk of foodborne parasite contaminations in circulating food in Putuo District, Shanghai Municipality, with relatively higher prevalence of foodborne parasite infections in marine fishes and raw pickled aquatic products of animal origin. The awareness of foodborne parasitic diseases knowledge is high among residents; however, the proportion of residents without behaviors of consuming raw or semi-raw food is low, indicating a discrepancy between the awareness and behaviors. Continuous monitoring and health education are required to be intensified.

Sequence characteristics and phylogenetic analysis of the ribosomal whole genome of Echinostoma miyagawai

CAO Yu, LI Ye, JIANG Botao

2025, 43(5):  697-704.  doi:10.12140/j.issn.1000-7423.2025.05.015

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Objective To identify the Echinostoma species from domestic ducks in Qiqihar City and analyze the characteristics and phylogenetic relationships of the Echinostoma ribosomal whole-genome sequences. Methods Echinostoma parasites were collected from the intestines of domestic ducks in Qiqihar City on July 2024, and the morphological characteristics of Echinostoma parasites were observed using the squash method. The DNA of the parasites was extracted and the internal transcribed spacer 2 (ITS2) sequence was amplified by PCR for parasites species identification, the ribosomal whole-genome sequence was amplified and subjected to sequence characteristics analysis. The sequencing results were aligned those recorded in the NCBI database using the BLAST tool to determine gene boundaries, and the presence of repetitive sequences was checked using a gene repeat sequence calculator. The secondary structure of the 18S, 5.8S and 28S was predicted using the RNAstructure software, and sequence similarity analysis was performed using the Megalign software. Sequence alignment was conducted using the MEGA 7.0, Clustal X, Paup, and Mrbayes software, and phylogenetic trees were constructed using the maximum parsimony method with 18S, ITS, and 28S sequences as marker genes. Results Three Echinostoma were isolated, which appeared a long leaf-like shape and had a well-developed head collar that contained 37 collar spines. The ITS sequence had a length of 1 037 bp, and the ITS2 sequence was 431 bp in length, which shared a 99.77% similarity with the ITS2 sequences of E. miyagawai isolated from Thailand (GenBank accession number: PQ821045) and Russia (GenBank accession number: OP696595), confirming the species as E. miyagawai. The ribosomal whole-genome sequence of E. miyagawai was 9 522 bp in length, and the lengths of the 18S, ITS1, 5.8S, ITS2, 28S and intergenic spacer (IGS) sequences were 1 989, 444, 162, 431, 3 858 and 2 638 bp, respectively. The 18S, ITS and 28S sequences were submitted to the GenBank database, with the accession number of PV455727; and the IGS sequence was submitted to the GenBase databases, with the accession number of C_AA117515.1. Repetitive sequences were found in ITS2, 28S, and IGS sequences, with the highest number of repetitive sequences seen in the 28S sequence. The rDNA sequence showed the highest similarity (99.3%) with the ribosomal genome sequence of E. miyagawai (GenBank accession number: OR509027). The secondary structure of 18S rRNA was composed of multiple stem-loop structures, which appeared a clover-like shape, and the secondary structure of 5.8S rRNA was relatively small and compact and formed a stable conformation via internal base pairs, while the secondary structure of 28S rRNA was complicated and appeared a multi-arm structure. Phylogenetic analysis revealed that phylogenetic trees were clustered into two clades, and the sequences in this study clustered on a small branch with those reported for E. miyagawai. Conclusion The Echinostoma species infecting domestic ducks in Qiqihar City is identified as E. miyagawai, and its ribosomal genome ITS2, 28S, and IGS sequences contain repetitive sequences and show close phylogenetic relationships with other E. miyagawai and E. revolutum from previous studies.

REVIEWS

Progress of researches on the role of IL-10 in the immunosuppressive microenvironment of alveolar echinococcosis

WANG Shujun, HAN Fei, ZHANG Huixia, ZHONG Shigui, ZHAO Yumin

2025, 43(5):  705-710.  doi:10.12140/j.issn.1000-7423.2025.05.016

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Alveolar echinococcosis (AE) is a zoonotic parasitic disease caused by infection with Echinococcus multilocularis larvae in hosts. As an exogenous substance, persistent larval E. multilocularis infections lead to suppression of cellular immune functions and dysregulation of cytokine secretion in hosts, thereby creating a unique immunosuppressive microenvironment in hosts. As a key anti-inflammatory cytokine, interleukin-10 (IL-10) plays a pivotal role in the immunomodulation of echinococcosis. This review presents the formation of the immunosuppressive microenvironment of AE, analyzes the biological characteristics of IL-10, its regulatory effects on immune cell functions, and its interplay with other cytokines, and discusses the mechanisms of the contribution of IL-10 to shaping the immunosuppressive microenvironment of AE, so as to provide new theoretical evidence for AE treatment and potential therapeutic targets of AE.

Progress of researches on interactions between sandfly gut microecology and Leishmania infection

HE Yaqi, CUI Lei, FANG Yuan, ZHOU Zhengbin, ZHANG Yi

2025, 43(5):  711-717.  doi:10.12140/j.issn.1000-7423.2025.05.017

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Sandflies, as vectors of multiples pathogens, are of significant epidemiological importance, and has been given attention due to their role in transmitting Leishmania. The growth and development of Leishmania is affected by the diverse microbial communities present in the sandfly gut, which also induces differential gene expression in the sandfly gut. This article summarizes the composition of the sandfly gut microbiota and their interactions with Leishmania, and alterations of functional characteristics in the sandfly gut caused by the parasite. Sandflies, Leishmania, gut microbiota, and gene expression form a complex network, and deciphering their interactions may provide more strategies for leishmaniasis management and sandfly vector control.

Current status and research progress of triclabendazole resistance in fascioliasis treatment

CHEN Chong, ZHU Aihong, HUANG Qin

2025, 43(5):  718-722.  doi:10.12140/j.issn.1000-7423.2025.05.018

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Fascioliasis, which caused by Fasciola hepatica and F. gigantica, is a zoonotic parasitic disease that not only affects human health but also poses a serious threat to other mammals, especially herbivores. Triclabendazole (TCBZ) is the first choice for fascioliasis treatment. However, economic data monitoring in health care indicates that the threat of TCBZ resistance has brought challenges to the health of animals in endemic areas and caused huge economic losses. At the same time, clinical reports have also documented treatment failures possibly related to TCBZ resistance, posing risks to the prevention and control of fascioliasis. This article focuses on the current situation of TCBZ treatment failure and/or resistance in fascioliasis, risk factors for TCBZ resistance, resistance mechanisms, progress in new drug development and exploration of new treatment methods, with the aim of providing references for further research on the prevention and control of fascioliasis.

SHORT COMMUNICATIONS

Epidemiological characteristics of scrub typhus in Changning County, Yunnan Province from 2014 to 2023

TIAN Yuxiu, HUANG Dongsheng

2025, 43(5):  723-726.  doi:10.12140/j.issn.1000-7423.2025.05.019

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Scrub typhus surveillance data in Changning County from January 1, 2014 to December 31, 2023 were collected from the China Disease Prevention and Control Information System, and a descriptive epidemiological analysis was performed using the software SPSS 27.0. Spatial autocorrelation analysis was performed, and Moran’s I was calculated. The concentration index (M value) of scrub typhus was calculated to analyze the seasonal characteristics of the disease. A total of 2 861 scrub typhus cases were reported in Changning County from 2014 to 2023, with an average annual incidence rate of 83.19/10⁵ and including one death. The incidence of scrub typhus increased from 30.69/10⁵ in 2014 to 166.97/10⁵ in 2023, which appeared an overall tendency towards a rise (χ²_trend = 553.555, P < 0.01). The peak incidence period is July to October each year, accounting for 91.26% (2 611/2 861) of reported cases. Concentration results showed that the highest M value was 0.905 in 2014, and the M values ​​for the remaining years ranged from 0.741 to 0.862, indicating that scrub typhus has a strong seasonal pattern. The scrub typhus cases were distributed across 13 townships in the county, the top five townships with the highest average annual reported incidence rates were Wengdu (353.10/10⁵), Mengtong (189.42/10⁵), Datianba (172.23/10⁵), Gengga (160.00/10⁵), and Jifei (103.97/10⁵). The average annual incidence rates in the remaining townships were all below 60/10⁵. The global spatial autocorrelation analysis indicated that for the period 2014-2023, all Moran’s I values were positive and statistically non-significant (P > 0.05). Males accounted for 49.91% (1 428/2 861), while females accounted for 50.09% (1 433/2 861). The cases distributed across all age groups, which were predominantly found at ages of 0 to 4 years (12.62%, 361/2 861) and 40 to 59 years (40.62%, 1 162/2 861), respectively. Farmers constituted the majority of the occupational distribution (74.00%, 2 117/2 861), and were primarily reported by township healthcare centers (52.25%, 1 495/2 861). These data demonstrate that the prevalence of scrub typhus showed an overall upward tendency in Changning County with a strong seasonality, distinct geographical and population distribution characteristics. Prevention and control efforts of scrub typhus should focus on rural areas during the transmission season.

Prevalence of intestinal parasitic infection in wild mice along the Yangtze River basins in Anhui Province in 2022

XU Xiaojuan, CHEN Xuefeng, WU Fan, WU Chenyang, LIU Ting, DAI Bo, ZHANG Shiqing

2025, 43(5):  727-730.  doi:10.12140/j.issn.1000-7423.2025.05.020

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The prevalence of parasitic infections was investigated in wild mice in 50 administrative villages across 38 townships in 6 cities of Wuhu, Ma’anshan, Tongling, Anqing, Chizhou, and Xuancheng in Anhui Province from October to November 2022. Wild mice were captured with baited traps or cages at night, and characterized for species. Then, mice were sacrificed, and the liver and mesenteric tissues were dissected and examined for parasitic infections using tissue biopsy or microscopy. Mouse stool samples were collected and tested for parasite eggs using the modified Kato-Katz thick smear technique. The differences in infection rates were analyzed using the χ² test in SPSS 23.0 software. A total of 1 703 wild mice were captured, and 678 were tested positive for parasitic infections, yielding an overall infection prevalence rate of 39.81%. The three cities with the highest prevalence included Xuancheng City (77.50%, 62/80), Wuhu City (72.97%, 54/74), and Anqing City (64.00%, 272/425). The captured mice included 931 females, 769 males, and 3 of unknown sex, and the infection rates were 40.92% (381/931), 38.49% (296/769) and 1/3, respectively, with comparable prevalence between male and female mice (χ² = 1.040, P > 0.05). Rattus losea (41.87%, 713/1 703), Apodemus agrarius (38.64%, 658/1 703) and R. tanezumi (11.27%, 192/1 703) were predominant mouse species, with prevalence rates of 34.64% (247/713), 40.58% (267/658) and 61.46% (118/192), respectively, and there was a mouse species-specific prevalence rate of parasitic infections (χ² = 48.484, P < 0.01). Parasites detected in the 678 infected rodents included hookworm, Ascaris lumbricoides, Capillaria hepatica, Hymenolepis nana, H. diminuta, Fasciolopsis buski, Trichuris trichura and Opisthorchis. Mono-infection accounted for 78.61% (533/678) of wild mice, and co-infections with two parasite species occurred in 20.21% (137/678), while triple infections with three parasite species occurred in 1.18% (8/678). The findings indicate that parasitic infections are highly prevalence in wild mice in Anhui Province, which poses a risk to human parasitic infections. Intensified monitoring of parasitic infections is recommended in wild mice.

Establishment and evaluation of modified Wright-Giemsa staining for detection of Blastocystis hominis

LIU Xiaoqing, ZOU Xiaohong, WANG Huiting, ZHOU Chen, LI Zhenhua, SHI Liang

2025, 43(5):  731-735.  doi:10.12140/j.issn.1000-7423.2025.05.021

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Fecal samples were collected from patients admitted to The Eighth Affiliated Hospital of Sun Yat-sen University from April 2021 to December 2024, and a total of 131 cases tested positive for Blastocystis hominis infections using PCR assay and 50 cases tested negative were randomly divided into the experimental group (92 cases tested positive, 35 cases tested negative) and validation group (39 cases tested positive, 15 cases tested negative) using a random number table. Each fecal sample of experimental group was prepared into suspensions, and 50, 100, and 150 μl of suspensions were sampled for preparation of thin smears. Compared to conventional Wright-Giemsa staining, which involves an additional drying step, they dried at 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, and 80 ℃. The smears were stained with Wright-Giemsa stains for 5 to 10 minutes and then observed under an optical microscope. The diagnostic performance of the modified Wright-Giemsa staining method was evaluated using PCR amplification and genotyping results with B. hominis-specific primers BhRDr/RD5 as a reference standard. The validation group used both the modified Wright-Giemsa staining method and PCR method for simultaneous detection of B. hominis. The sensitivity, specificity, Youden index, predictive values (positive/negative), kappa coefficient, and concordance rate were as reference standards to evaluate the diagnostic performance of the modified method. The optimal fecal smear volume was 50 μl, and the most suitable drying temperature was (70 ± 5) ℃. PCR amplification produced a specific band at approximately 600 bp, with ST3 as the predominant genotype, followed by ST7, ST1, ST6, and ST2. The diagnostic sensitivity, specificity, kappa coefficient and concordance rate of the modified Wright-Giemsa staining method were 98.9% (95% CI: 94.0% to 100.0%), 97.1% (95% CI: 85.3% to 99.9%), 0.961 (95% CI: 0.906 to 1.000), and 98.4% (125/127) in the experimental group, showing high consistency with PCR assay. McNemar’s test was used to compare the differences in detection results between the two methods in the validation group, and the differences were not statistically significant (P > 0.05). The diagnostic sensitivity, specificity, positive predictive value, negative predictive value, Youden index, kappa coefficient and concordance rate of the modified Wright-Giemsa staining method were 97.4% (95% CI: 86.2% to 99.9%), 86.7% (95% CI: 59.5% to 98.3%), 95.0% (95% CI: 83.8% to 99.5%), 92.9% (95% CI: 69.8% to 99.8%), 0.841 (95% CI: 0.662 to 1.000), 0.858 (95% CI: 0.703 to 1.000), and 94.4% (51/54), respectively. The modified Wright-Giemsa staining method established in this study is simple to operate, yields stable results, and shows high consistency with PCR assay, which is suitable for rapid screening of B. hominis in grassroots medical institutions and field epidemiological investigations.

CASE REPORT

Strongyloides stercoralis and Pneumocystis jirovecii co-infection in the lung: A case report

JIA Dan, XIE Tingyi, TANG Mingwen, LI Yuzhen, HE Yongshi, WANG Yaqiong, HUANG Zhilan, XIE Wei

2025, 43(5):  736-739.  doi:10.12140/j.issn.1000-7423.2025.05.022

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In May 3, 2025, a 59-year-old female patient was admitted to Shenzhen Traditional Chinese Medicine Hospital with complaints of fatigue and shortness of breath for over one month, worsened with cough and expectoration for 10 days. The case was engaged in agriculture for a long period of time, and worked in a restaurant 3 months prior to admission, with exposure to fresh food. Admission examinations showed a white blood cell count of 5.60 × 10⁹/L, a neutrophils proportion of 89.6%↑, an eosinophil count was 0.02 × 10⁹/L, and blood gas analysis revealed a pH value of 7.438, CO₂ partial pressure (PCO₂) of 35.5 mmHg, O₂ partial pressure (PO₂) of 57 mmHg↓, and oxygenation index of 173 mmHg↓. Pulmonary function tests revealed moderate to severe mixed ventilatory dysfunction with a positive bronchodilation test. Chest CT scans displayed diffuse mosaic and ground-glass opacities in bilateral lungs. Strongyloides stercoralis was found in sputum smears and bronchoalveolar lavage fluid under a bronchoscope. Targeted sequencing of bronchoalveolar lavage fluid under a bronchoscope identified 75 545 sequences of Pneumocystis jirovecii, and targeted sequencing of bloodstream infections identified 276 631 sequences of P. jirovecii, with abundance of 99.38%. The case was diagnosed as S. stercoralis and P. jirovecii co-infection in the lung, and was given oral administration of albendazole (0.4 g/d) and compound sulfamethoxazole tablets (0.4 g, 80 mg/6 h), along with anti-inflammatory and anti-asthmatic therapy. However, the patient’s disease condition continued to progress, with worsening dyspnea and remarkable chest tightness, and the case was promptly transferred to a specialized hospital for further management in May 9. Follow-up revealed that the infection continued to worsen after transfer to another hospital, endotracheal intubation, ventilator and ECMO support were given.