Table of Content

25 April 2025, Volume 37 Issue 2

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Strategic thinking on management of cross⁃boundary imported schistosomiasis

XU Jing, LI Shizhen, LI Qin, GUO Suying, LI Shizhu, ZHOU Xiaonong

2025, 37(2):  107-111. 

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Schistosomiasis is prevalent in 78 countries and territories worldwide, while the eastern and western parts of sub⁃Sahara Africa bear the highest disease burden due to schistosomiasis. Recently, climate change, international trade and travel, urbanization and war have increased the risk of cross⁃boundary importation and transmission of schistosomiasis, and schistosomiasis has increasingly become a public health concern in non⁃endemic countries and territories. Biomphalaria straminea, the intermediate host of Schistosoma mansoni, has colonized in southern China and its habitats continue to move northward. In addition, cross⁃boundary imported cases of schistosomiasis have been reported occasionally in China. However, the real number of cases may be underestimated greatly due to insufficient diagnostic capacity and weak awareness of case reporting for overseas imported schistosomiasis in healthcare facilities. It is necessary to establish a multi⁃party collaborative mechanism, improve corresponding systems and technical specifications, reinforce surveillance and early warning, and border management, enhance technical reserves and capability building, and improve the awareness of schistosomiasis prevention and healthcare⁃seeking among entry⁃exit personnel, in order to effectively address the threat of cross⁃boundary imported schistosomiasis.

The future is already here and the wave is coming: progress and challenges in the research and application of artificial intelligence-empowered malaria control programmes

ZHU Guoding, CAO Jun

2025, 37(2):  112-115. 

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The rapid rise and fast development of artificial intelligence (AI) has brought unprecedented opportunities and challenges to all sectors, including disease prevention control. Malaria is one of the world's most devastating infectious diseases. This article summarizes the advances in the research and application of AI⁃empowered malaria control programmes, analyzes key challenges during the implementation of malaria control programmes, and proposes future development directions and research proprieties, so as to provide insights into facilitating the translation of AI⁃driven strategies in global infectious disease control efforts.

Sequencing and analysis of the complete mitochondrial genome of Bulinus globosus

QIAN Peijun, Masceline Jenipher Mutsaka⁃Makuvaza, LÜ Chao, QIAN Yingjun, WANG Wenya, CHEN Shenglin, XU Andong, XUE Jingbo, XU Jing, ZHOU Xiaonong, Nicholas Midzi, LI Shizhu

2025, 37(2):  116-126, 135. 

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Objective　To analyze the structural and phylogenetic characteristics of the mitochondrial genome from Bulinus globosus, so as to provide a theoretical basis for classification and identification of species within the Bulinus genus, and to provide insights into understanding of Bulinus⁃schistosomes interactions and the mechanisms of parasite transmission.  Methods　       B. globosus samples were collected from the Ruya River basin in Zimbabwe. Mitochondrial DNA was extracted from B. globosus samples and the corresponding libraries were constructed for high⁃throughput sequencing on the Illumina NovaSeq 6000 platform. After raw sequencing data were subjected to quality control using the fastp software, genome assembly was performed using the A5⁃miseq and SPAdes tools, and genome annotation was conducted using the MITOS online server. Circular maps and sequence plots of the mitochondrial genome were generated using the CGView and OGDRAW software, and the protein conservation motifs and structures were analyzed using the TBtools software. Base composition and codon usage bias were analyzed and visualized using the software MEGA X and the ggplot2 package in the R software. In addition, a phylogenetic tree was created in the software MEGA X after sequence alignment with the software MAFFT 7, and visualized using the software iTOL.  Results　         The mitochondrial genome of B. globosus was a 13 730 bp double⁃stranded circular molecule, containing 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 13 protein⁃coding genes, with a marked AT preference. The mitochondrial genome composition of B. globosus was similar to that of other species within the Bulinus genus. Phylogenetic analysis revealed that the complete mitochondrial genome sequence of B. globosus was clustered with B. truncatus, B. nasutus, and B. ugandae into the same evolutionary clade, and gene superfamily analysis showed that the metabolism⁃related proteins of B. globosus were highly conserved, notably the cytochrome c oxidase family, which showed a significant consistency.  Conclusions　This is the first whole mitochondrial genome sequencing to decode the compositional features of the mitochondrial genome of B. globosus from Zimbabwe and its evolutionary relationship within the Bulinus genus, which provides important insights for further understanding of the phylogeny and mitochondrial genome characteristics of the Bulinus genus.

Effects of Toxoplasma gondii type Ⅰ/Ⅱ rhoptry protein 16 on the polarization and inflammatory response of mouse alveolar macrophages

LI Jiaming, DANG Tiantian, ZHAO Zhijun

2025, 37(2):  127-135. 

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Objective　To investigate the effects of Toxoplasma gondii typeⅠand Ⅱrhoptry protein 16 (ROP16) on the polarization and inflammatory response of mouse alveolar macrophages, so as to provide the scientific evidence for unveiling the immunoregulatory mechanisms following T. gondii infection in host cells and the clinical diagnosis and treatment of pulmonary toxoplasmosis. Methods　Mouse alveolar macrophages served as blank controls, and mouse alveolar macrophages transfected with the empty lentiviral expression vector served as negative controls, and mouse alveolar macrophages transfected with lentiviral vectors overexpressing T. gondii typeⅠand Ⅱ ROP16 served as the typeⅠand ⅡROP16 overexpression groups. Following puromycin selection, stably transfected cells that overexpressed typeⅠand ⅡROP16 were generated, observed for green fluorescence expression under a fluorescence microscope and verified using PCR, Western blotting and real⁃time quantitative reverse transcription PCR (RT⁃qPCR) assays. The expression of ROP16, inducible nitric oxide synthase (iNOS), arginase (Arg)⁃1, mannose receptor (CD206), cluster of differentiation 86 (CD86), NOD⁃like receptor thermal protein domain associated protein 3 (NLRP3), caspase⁃1, apoptosis⁃associated speck⁃like protein containing a CARD (ASC), and interleukin (IL)⁃1β proteins was determined in mouse alveolar macrophages using Western blotting assay, and the mRNA levels of ROP16, iNOS, IL⁃1β, IL⁃4, IL⁃12, IL⁃18, Arg⁃1, IL⁃10, IL⁃6, tumor necrosis factor (TNF)⁃α and transforming growth factor (TGF)⁃β were detected in mouse alveolar macrophages using RT⁃qPCR assay. Results　Fluorescence microscopy showed 90% of mouse alveolar macrophages producing green fluorescent signals in the typeⅠand Ⅱ ROP16 overexpression groups and the negative control group. The relative ROP16 protein expression was  1.000 ± 0.000, 1.003 ± 0.020, 1.349 ± 0.055, and 1.376 ± 0.080 in mouse alveolar macrophages in the blank control group, negative control group, and typeⅠand ⅡROP16 overexpression groups (F = 35.30, P < 0.01), and the relative ROP16 mRNA expression was 1.007 ± 0.172, 2.030 ± 0.356, 1 409.579 ± 75.960, and 1 413.581 ± 27.712 in the blank control group, negative control group, and typeⅠand Ⅱ ROP16 overexpression groups (F = 811.00, P < 0.01). The ROP16 expression was significantly higher in the typeⅠand ⅡROP16 overexpression groups than in the blank control group at both protein and mRNA levels (all P value < 0.01). Western blotting assay detected significant differences among the four groups in terms of iNOS, Arg⁃1, CD86, CD206, NLRP3, caspase⁃1, ASC, and IL⁃1β protein expression (F = 124.70, 82.40, 79.82, 919.40, 84.74, 39.85, 2 354.00 and 65.96, all P values < 0.05), and the expression of Arg⁃1, CD206, NLRP3, caspase⁃1, ASC, and IL⁃1β proteins was significantly higher in the typeⅠROP16 overexpression group than in the blank control group (all P values < 0.001), while the expression of iNOS, CD86, NLRP3, caspase⁃1, ASC, and IL⁃1β proteins was significantly higher in the type ⅡROP16 overexpression group than in the blank control group (all P values < 0.01). RT⁃qPCR assay detected significant differences among the four groups in terms of iNOS, IL⁃1β, IL⁃4, IL⁃12, IL⁃18, Arg⁃1, IL⁃10, IL⁃6, TNF⁃α, and TGF⁃β mRNA expression (F = 407.00, 1 528.00, 833.10, 267.90, 989.80, 161.80, 461.10, 5 529.00, 849.60 and 8 836.00, all P values < 0.05), and the Arg⁃1, IL⁃4, IL⁃10, and TGF⁃β mRNA expression was significantly higher in the typeⅠROP16 overexpression group than in the blank control group (all P values < 0.001), while the iNOS, IL⁃1β, IL⁃12, IL⁃18, IL⁃6, and TNF⁃α mRNA expression was significantly higher in the typeⅡROP16 overexpression group than in the blank control group (all P values < 0.001). Conclusions　T. gondii typeⅠROP16 may induce M2⁃dominant phenotypes of mouse alveolar macrophages, and typeⅡROP16 may induce M1⁃dominant phenotypes of mouse alveolar macrophages. Both T. gondii typeⅠand ⅡROP16 may activate NLRP3, and mediate the activation of ASC, caspase⁃1 and IL⁃1β to promote inflammatory responses.

Proportions of memory T cells and expression of their associated cytokines in lymph nodes of mice infected with Echinococcus multilocularis

LI Yinshi, ADILAI Duolikun, DENG Bingqing, ABIDAN Ainiwaer, SUN Sheng, XIAO Wenying, GE Conghui, TANG Na, LI Jing, WANG Hui, JIANG Tao, ZHANG Chuanshan

2025, 37(2):  136-143,189. 

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Objective　To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolar echinococcosis.  Methods　Twenty⁃four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post⁃infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post⁃infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry.  Results　HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post⁃infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4+ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4+ tissue⁃resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post⁃infection, and higher proportions of CD4+ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764，P < 0.05] and CD4+ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039，P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post⁃infection. The proportions of CD8+ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post⁃infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)⁃α+ CD4+ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF⁃α+ CD8+ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF⁃α+ CD8+ Tm cells [7.0% (1.0%)  vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post⁃infection. Conclusions　Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8+ Tm cells to secrete the effector molecule TNF⁃α in mouse lymph nodes at the late⁃stage infection may facilitate chronic parasitism of E. multilocularis.

Population genetic structure of sandflies in China based on mitochondrial DNA

LI Zhongqiu, WEI Zixin, ZHOU Zhengbin, ZHANG Yi

2025, 37(2):  144-151. 

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Objective　To investigate the genetic diversity of sandfly populations in endemic areas of visceral leishmaniasis in China, so as to provide references insights into management of visceral leishmaniasis and the vector sandflies. Methods　Sixteen sampling sites were selected from main endemic foci of visceral leishmaniasis in China from June to September 2024, including Shanxi Province, Shaanxi Province, Henan Province, Gansu Province, Sichuan Province, and Xinjiang Uygur Autonomous Region. Sandflies were captured using light traps and manual aspirators from sheep pens, chicken coops, cave dwellings, bovine sheds, and pig pens at each sampling site. A single sandfly sample was washed in phosphate⁃buffered saline (PBS), and genomic DNA was extracted from sandfly samples. Cytochrome oxidase subunit 1 (COI) gene was amplified using PCR assay with universal primers, and analyzed and retrieved with the nucleotide sequence analysis tool (BLAST) software, and the sequence of COI gene was aligned with the ClustalX 1.83 and MEGA 7.0 software. The base composition and variation site of the COI gene sequence were analyzed using the software MEGA 7.0, and the number of haplotypes, total number of segregating sites, haplotype diversity, nucleotide diversity, and average nucleotide differences were calculated in the COI gene sequence using the software DnaSP 5.10, followed by Tajima's D test for neutrality. Haplotypes were screened using the software DnaSP 5.10, and the haplotype network map of sandfly samples was plotted using the software Network 5.0. MEGA 7.0 software was employed for gene sequence editing and alignment, and calculation of genetic distances among sandfly species sampled from different regions, and a phylogenetic tree was built with a neighbor⁃joining method. Results　A total of 466 sandflies were captured from 16 sampling sites in China from June to September 2024, and 430 gene sequences were yielded following PCR amplification and sequencing of the COI gene, with 652 to 688 bp in the length of amplification fragments. The captured sandfly samples were characterized as Phlebotomus chinensis, Sergentomyia squamirostris, Se. koloshanensis, Ph. sichuanensis, and Ph. longiductus following the COI gene sequence alignment in BLAST. A total of 251 haplotypes were identified in the 430 gene sequences from sandfly samples (50.5%), and the average haplotype diversity, nucleotide diversity and average number of nucleotide difference were 0.885, 0.257 and 160.761, respectively. The Tajima's D values were -0.92 for sandfly populations from Yangquan City,  Shanxi Province and -1.73 for sandfly populations from Sanmenxia City, Henan Province, and were all more than 0 for sandfly populations from other sampling sites. Haplotype analysis identified 50 haplotypes, which were classified into two haplogroups. Heplogroup 1 included 29 haplotypes, which had a high homology, and heplogroup 2 included 21 haplotypes. The average genetic distance was 0.000 to 0.604 among sandfly samples from different sampling sites, and phylogenetic analysis revealed that the five sandfly species were clustered into distinct clades, all with 100% clade confidence. Conclusions　There is a high genetic polymorphism in the COI gene from five sandfly populations in main endemic foci of visceral leishmaniasis in China, and COI gene may serve as a marker gene for analysis of the genetic structure of sandfly populations.


Non⁃coding RNAs expression profile of adjacent and distant liver tissues of hepatic cystic echinococcosis lesions

IRSHAT Ibrahim, AIZEMAITI Aikebaier, WUBULIKASIMU Mijiti, XU Qilin, ABUDUSIKUER Abudumijiti, WU Yuanquan, KAHAER Tuersun

2025, 37(2):  152-162. 

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Objective　To analyze the differential expression of non⁃coding RNAs (ncRNAs) from liver tissues adjacent to hepatic cystic echinococcosis (CE) lesions and distant normal liver tissues using whole transcriptome sequencing, and perform functional annotations of differentially expressed ncRNAs, so as  to explore the potential role of ncRNAs in the pathogenesis of CE. Methods　Intraoperative liver tissue specimens adjacent to hepatic CE lesions and distant normal liver tissue specimen were sampled from patients with hepatic CE, and the expression profiles of microRNAs (miRNAs), circular RNAs (circRNAs), and long non⁃coding RNAs (lncRNAs) were detected using whole transcriptome sequencing. Differentially expressed genes were identified, and functional annotations were performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. In addition, a circRNA/lncRNA⁃miRNA⁃messenger RNA (mRNA) competing endogenous RNA (ceRNA) network was constructed using the Cytoscape software, and the expression of hub miRNAs in the network was validated using real⁃time quantitative reverse transcription PCR (RT⁃qPCR) assay.  Results　A total of 41 differentially expressed miRNAs were identified between the adjacent and distal tissues of hepatic CE lesions, including 8 up⁃regulated and 33 down⁃regulated miRNAs, which were significantly enriched in biological processes of Ras signaling and neutrophil activation. Five differentially expressed circRNAs were detected, including 3 up⁃regulated and 2 down⁃regulated circRNAs, which were significantly enriched in molecular functions of hormone signaling pathways and RNA transcription regulation. A total of 447 differentially expressed lncRNAs were identified, including 200 up⁃regulated and 247 down⁃regulated lncRNAs, which were involved in cell proliferation, immune regulation, and extracellular matrix remodeling pathways. MiRNA target analysis predicted hsa⁃miR⁃27a⁃5p, hsa⁃miR⁃21⁃3p, and hsa⁃miR⁃181b⁃2⁃3p as hub nodes in the ceRNA network. RT⁃qPCR assay detected that the relative expression levels of ENSG00000253736, HAS2⁃AS1, PCSK6, hsa⁃miR⁃21⁃3p, hsa⁃miR⁃27a⁃5p, MIR23AHG, VIPR1⁃AS1, LINC02910, and hsa⁃miR⁃181b⁃2⁃3p were 3.00 ± 0.25, 2.75 ± 0.33, 1.01 ± 0.51, 2.65 ± 0.41, 1.01 ± 0.29, 1.10 ± 0.31, 1.05 ± 0.27, 0.25 ± 0.49, and 2.56 ± 0.35 in adjacent tissues of hepatic CE lesions, normalized to that in distant tissues from hepatic CE lesions,  respectively (t = 6.21, 5.83, 7.51, 7.46, 6.12, 6.65, 7.13, 1.87 and 7.81, all P values  < 0.01), which was consistent with whole transcriptome sequencing results. Conclusions　Differentially expressed ncRNAs from adjacent and distal liver tissues of hepatic CE lesions may contribute to the pathological mechanisms of CE through mediating cell proliferation, immune evasion, and inflammatory responses, in which hsa⁃miR⁃27a⁃5p and hsa⁃miR⁃21⁃3p may serve as hub miRNAs.

Dynamics of eosinophil infiltration and microglia activation in brain tissues of mice infected with Angiostrongylus cantonensis

WEI Fanna, ZHANG Renjie, HU Yahong, QIN Xiaoyu, GUO Yunhai, MO Xiaojin, LU Yan, SUN Jiahui, ZHOU Yan, GUO Jiatian, SONG Peng, CHU Yanhong1, XU Bin, ZHANG Ting, CAI Yuchun, CHEN Muxin

2025, 37(2):  163-175. 

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Objective　To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods　Fifty BALB/c mice were randomly divided into the control group and the 7⁃d, 14⁃d, 21⁃day and 25⁃d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage Ⅲ A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post⁃infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post⁃infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor Ⅲ (Fcgr3), Fcγ receptor Ⅱb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C⁃type 1 (Mrc1), chitinase⁃like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real⁃time quantitative reverse transcription PCR (RT⁃qPCR) assay in the mouse cerebral cortex of mice post⁃infection. Results　A large number of A. cantonensis larvae were seen on the mouse meninges surface post⁃infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7⁃d, 14⁃d, 21⁃d and 25⁃d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7⁃d, 14⁃d, 21⁃d and 25⁃d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14⁃d (3.08 ± 0.78) and 21⁃d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi⁃quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14⁃d (5.75 ± 1.28), 21⁃d (6.23 ± 1.89) and 25⁃d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells [(1.30 ± 0.01) vs. (1.41 ± 0.03); t = 5.266, P < 0.05] were reduced in mouse brain tissues in the 21⁃d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05) , Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.123, P < 0.05) in mice brain tissues. Conclusions　A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions. 

Effectiveness of the integrated schistosomiasis control programme in Wuhan City from 2005 to 2023

WANG Shuai, LUO Huatang, LI Yang, WANG Hao, LIU Cong, XIONG Yuelin, ZHANG Jiajing, ZHU Wen

2025, 37(2):  176-183,195. 

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Objective　To evaluate the effectiveness of the integrated schistosomiasis control programme in Wuhan City from 2005 to 2023, so as to provide insights into precision control and elimination of schistosomiasis.  Methods　The integrated measures for schistosomiasis control implemented by health, agriculture, water resources, and forestry departments of Wuhan City, and the epidemiological data of schistosomiasis in Wuhan City were collected from 2005 to 2023, and the prevalence of human schistosomiasis, prevalence of Schistosoma japonicum infections in humans and bovines, areas of S. japonicum⁃infected snail habitats, areas of snail habitats in inner embankments, and actual areas of snail habitats were retrieved. In addition, the trends in prevalence of schistosomiasis in humans and livestock and snail status were evaluated in Wuhan City from 2005 to 2023 using Mann⁃Kendall test and a Joinpoint regression model.  Results　Mann⁃Kendall test revealed a tendency towards a decline in the prevalence of human schistosomiasis (Z = -4.41, P < 0.01), prevalence of S. japonicum infections in humans (Z = -4.89, P < 0.01) and bovines (Z = -4.50, P < 0.01), areas of S. japonicum⁃infected snail habitats (Z = -3.91, P < 0.01), areas of snail habitats in inner embankments (Z = -2.28, P = 0.02), and actual areas of snail habitats (Z = -5.95, P < 0.01) in Wuhan City from 2005 to 2023. Joinpoint regression analysis showed an average annual reduction of 8.58% in the prevalence of human schistosomiasis in Wuhan City from 2005 to 2023 [average annual percent change (AAPC) = -8.58%, 95% confidence interval (CI): (-10.02%, -6.65%), P < 0.01], with two joinpoints in 2013 and 2016, respectively, and the tendency towards a decline showed statistical significance during the period from 2013 through 2016 [annual percent change (APC) = -34.41%, 95% CI: (-40.36%, -20.01%), P < 0.01]. The prevalence of S. japonicum human infections appeared an average annual reduction of 51.91% in Wuhan City from 2005 to 2023 [AAPC = -51.91%, 95% CI: (-58.12%, -44.25%), P < 0.01], with two joinpoints in 2014 and 2017, respectively, and the tendency towards a decline showed statistical significance during the period from 2014 through 2017 [APC = -98.17%, 95% CI: (-99.17%, -90.87%), P < 0.01]. The prevalence of S. japonicum infections in bovines appeared an average annual reduction of 53.12% in Wuhan City from 2005 to 2023 [AAPC = -53.12%, 95% CI: (-59.65%, -42.44%), P < 0.01], with two joinpoints in 2011 and 2014, respectively, and the tendency towards a decline showed statistical significance during the period from 2014 through 2017 [APC = -98.63%, 95% CI: (-99.44%, -90.93%), P < 0.01]. The areas of S. japonicum⁃infected snail habitats appeared an average annual reduction of 47.09% in Wuhan City from 2005 to 2023 [AAPC = -47.09%, 95% CI: (-52.92%, -38.26%), P < 0.01], with two joinpoints in 2011 and 2014, respectively, and the tendency towards a decline showed statistical significance during the period from 2011 through 2014 [APC = -97.27%, 95% CI: (-98.65%, -88.06%), P < 0.01]. The areas of snail habitats in inner embankments appeared an average annual reduction of 4.45% in Wuhan City from 2005 to 2023 [AAPC = -4.45%, 95% CI: (-5.18%, -3.82%), P < 0.01], with three joinpoints in 2011, 2015 and 2018, respectively, and statistical significance was seen in the tendency towards a decline during the period from 2005 through 2011 [APC = -16.38%, 95% CI: (-20.15%, -14.25%), P < 0.01]. In addition, the actual areas of snail habitats appeared an average annual reduction of 2.65% in Wuhan City from 2005 to 2023 [AAPC = -2.65%, 95% CI: (-2.89%, -2.40%), P < 0.01], with a joinpoint in 2013, and the tendency towards a decline showed statistical significance during the period from 2013 through 2023 [APC = -4.06%, 95% CI: (-4.66%, -3.58%), P < 0.01]. Conclusions　The integrated schistosomiasis control programme achieved significant effectiveness in Wuhan City from 2005 to 2023, with a tendency towards a decline in morbidity due to schistosomiasis in humans and livestock and snail status. The integrated schistosomiasis control strategy with emphasis on management of the source of S. japonicum infections should continue to be implemented to consolidate the schistosomiasis control achievements and achieve the goal of schistosomiasis elimination in the city.

Investigation of tick species in Suizhou City, Hubei Province from 2023 to 2024

LU Huiya, GUO Fang, PAN Yibin, PENG Meng, WU Libang, LIN Ye, LIU Xiaohui, YU Xuejie

2025, 37(2):  184-189. 

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Objective　To investigate the species of ticks in Suizhou City, Hubei Province, so as to provide insights into management of ticks and tick⁃borne diseases. Methods　During the period between May 2023 and June 2024, livestock breeding farms and vegetation neighboring the place of residence of confirmed and suspected patients with tick⁃borne disease were selected as sampling points in rural areas from Yindian Township, Gaocheng Township, Wanhe Township, Wushan Township, Xiaolin Township, Xihe Township, Hedian Township and Beijiao Street in Suizhou City, Hubei Province, where confirmed and suspected cases with tick⁃borne diseases had been reported. The parasitic ticks on the body surface of free⁃range livestock were captured with tweezers in livestock breeding farms, and free ticks on the vegetation surface were captured with the flagging method. Morphological identification of tick samples was performed under a microscope, and the gender and developmental stage of ticks were determined. One engorged adult tick, 2 to 3 blood⁃feeding but non⁃engorged adult ticks, 10 to 15 unfed female ticks, 15 to 20 unfed male ticks, and 30 to 40 tick nymphs or larvae were assigned into a group, respectively. Genomic DNA was extracted from tick samples in each group, and mitochondrial 16S rRNA gene was amplified. Sequence analysis was performed with the DNASTAR software, and phylogenetic analysis was performed using the software MEGA 7.0. In addition, the phylogenetic tree was generated using the maximum likelihood method based on the Kimura 2 parameter model. Results　A total of 2 438 ticks were captured from Suizhou City, Hubei Province during the period between May 2023 and June 2024, including 595 free ticks and 1 483 parasitic ticks. Three developmental stages of ticks were captured, including larvae, nymphs, and adults, and 75.18% (1 899/2 438) of captured ticks were adult, in which 79.04% (1 501/1 899) were female. Morphological and molecular biological assays identified one family, three genera and four species of captured ticks, including 2 425 Haemaphysalis longicornis ticks (99.47%) and one H. flava tick (0.04%) of the genus Haemaphysalis, 11 Rhipicephalus microplus ticks (0.45%) of the genus Rhipicephalus, and one Ixodes sinensis tick (0.04%) of the genus Ixodes in the family Ixodidae. Phylogenetic analysis revealed that the H. longicornis sequence (SZ49) in this study was clustered with sequences from Yunnan Province (GenBank accession number: MH024510.1), Hebei Province (GenBank accession number: MK450606.1) and Henan Province (GenBank accession number: MZ230645.1) into a clade, and the H. flava sequence (SZ19) in this study was clustered with sequences from Japan (GenBank accession number: MW064044.1), South Korea (GenBank accession number: ON629585.1), and Jiangsu Province (GenBank accession number: PP494741.1) and Hebei Province of China (GenBank accession number: MH520685.1) into a clade, while the R. microplus sequence (SZ8) in this study was clustered with the sequences from India (GenBank accession number: MK621328.1), and Henan Province (GenBank accession number: MT555307.1) and Guizhou Province of China (GenBank accession number: PP446801.1) into a clade. The sequence of I. sinensis (SZ23) in this study had 99.51% homology with that (GenBank accession number: OM368265.1) of ticks sampled from Wuhan City, Hubei Province. Conclusion　There are four tick species of H. longicornis, H. flava, R. microplus and I. sinensis in Suizhou City, Hubei Province, and H. longicornis is the dominant species. H. flava is firstly recorded in Suizhou City. 

Risk assessment of human Spirometra mansoni infections and cross⁃sectional study on knowledge, attitude and practice towards sparganosis in endemic areas of Henan Province

ZHANG Yalan, JIANG Tiantian, MA Xiaohui, DENG Yan, CHEN Weiqi, ZHU Yankun, TANG Zhenqiang, LIN Ximeng, ZHANG Hongwei

2025, 37(2):  190-195. 

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Objective　To assess the risk of human Spirometra mansoni infections and investigate the knowledge, attitude and practice (KAP) towards sparganosis mansoni among residents in Henan Province, so as to provide insights into formulation of the sparganosis mansoni control measures. Methods　Qinling Village in Fugou County of Zhoukou City, Bali Village in Yancheng District of Luohe City, Duzhai Village in Puyang County of Puyang City and Doushan Village in Luoshan County of Xinyang City were sampled as survey sites in Henan Province from July to August 2023, and more than 40 frogs were sampled from ponds or streams in each survey site for detection of Sparganum mansoni infections. At least 150 residents were sampled using a cluster sampling method from each survey site, and the sero⁃prevalence of anti⁃S. mansoni IgG antibody was estimated. In addition, a questionnaire survey was conducted on the KAP towards sparganosis mansoni among participants, and the proportion of eligible KAP, rate of correct KAP and KAP scores were calculated.  Results　A total 229 frogs were collected from 4 survey sites in 2023, and the overall prevalence of S. mansoni infection was 4.37% (10/229) in frogs, with 7.75% (10/129) prevalence in wild frogs and 0 in farm⁃bred frogs. A questionnaire survey was performed among 649 residents sampled from 4 survey sites, and 649 serum samples were collected. The sero⁃prevalence of anti⁃S.mansoni IgG antibody was 0.15% (1/649) and the overall proportion of eligible KAP was 23.73% (154/649) among participants. There were age⁃ ([χ2]  = 30.905, P = 0.000), educational level⁃ ([χ2] = 41.011, P = 0.000), and occupation⁃specific proportions of eligible KAP among participants ([χ2] = 10.721, P = 0.005), and the proportion of eligible KAP decreased with age ([χ2] trend = 22.717, P = 0.000) and increased with education levels ([χ2] trend = 40.025, P = 0.000). The rates of correct KAP towards sparganosis mansoni were 40.81% (2 119/5 192), 96.66% (1 882/1 947) and 63.81% (3 727/5 841) ([χ2] = 1 913.731, P = 0.000) among residents, respectively. The rates of correct KAP towards sparganosis mansoni varied significantly among survey sites ([χ2] = 136.872, 42.347 and 255.157; all P values= 0.000, with the highest rate of correct knowledge (51.94%, 748/1 440) and practices (75.86%, 1 229/1 620) in Yancheng District of Luohe City and the highest rate of correct attitudes in Puyang County of Puyang City (99.11%, 446/450) (all P values< 0.05).  Conclusions　There is still a high transmission risk of sparganosis mansoni in Henan Province, and the KAP towards sparganosis mansoni is required to be improved among residents.

Toxicity and sublethal effects of calcium cyanamide against susceptible strains of Aedes albopictus

ZHENG Luyang, XU Huiyi, WEN Qingqiu, ZHOU Ning, ZHENG Xueli

2025, 37(2):  196-200,222. 

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Objective　To examine the toxicity and sublethal effects of calcium cyanamide against susceptible isolates of Aedes albopictus, so as to provide insights into rational use of calcium cyanamide for integrated management of Ae. albopictus. Methods　The sublethal concentrations ［30% lethal concentration (LC30) and median lethal concentration (LC50)］of calcium cyanamide against susceptible strains of Ae. albopictus were determined using the larval immersion test. With 100 mL of dechlorinated water as the control group, after the larvae of susceptible strains of Ae. albopictus were immersed in calcium cyanamide for 24 hours, the pupation rate, pupation duration, emergence rate, number of eggs laid, percentage of eggs hatched, and lifespan of Ae. albopictus were calculated and compared post⁃treatment with calcium cyanamide at different sublethal concentrations. The midgut tissues of larvae of susceptible strains of Ae. albopictus treated with 100 mg/L calcium cyanamide were sampled for pathological sectioning to observe midgut tissue damages. To evaluate the residual activity, 100 larvae of susceptible strains of Ae. albopictus were treated with 200 mg/L and 500 mg/L calcium cyanamide, and the mortality of larvae was calculated every 24 hour, with dead larvae replaced until no larval death. Results　The regression equation for the toxicity of calcium cyanamide against larvae of susceptible strains of Ae. albopictus was y = -9.441 + 4.657x, with an LC50 of 106.42 mg/L［95% confidence interval (CI): (94.64, 118.36) mg/L］and an LC30 of 82.17 mg/L［95% CI: (94.64, 118.36) mg/L］, respectively. After larvae of susceptible strains of Ae. albopictus were treated with sublethal concentrations (LC30 and LC50) of calcium cyanamide for 24 hours, there were reduced pupation and emergence rates of larvae (all P values < 0.000 1), prolonged pupal stage (both P values < 0.000 1), reduced numbers of eggs laid by survival female Ae. albopictus (both P values < 0.000 1), reduced percentages of eggs hatched by Ae. albopictus eggs (both P values < 0.000 1), and reduced median survival period of survival female Ae. albopictus ([χ2] = 9.36 and 20.33, both P values < 0.01) in the LC30 and LC50 groups relative to the control group. There was a numerical decline in the median survival period of survival female Ae. albopictus in the LC30 groups relative to the control group ([χ2] = 2.42, P > 0.05), and there was a significant decline in the median survival period of survival female Ae. albopictus in the LC50 group relative to the control group ([χ2] = 11.42, P < 0.01). Histopathological examinations showed severe damages to the midgut tissues of larvae of susceptible strains of Ae. albopictus, and residual activity assay revealed that the mortality of larvae of susceptible strains of Ae. albopictus was both 0 on day 32 post⁃treatment with calcium cyanamide at a concentration of 200 mg/L and on day 70 post⁃treatment with calcium cyanamide at a concentration of 500 mg/L, showing complete loss of the larvicidal activity of calcium cyanamide.  Conclusions　Calcium cyanamide is highly toxic against susceptible strains of Ae. albopictus, and calcium cyanamide at sublethal concentrations (LC30 and LC50) may inhibit growth, development, and reproductive capability of susceptible strains of Ae. albopictus, and shorten the lifespan of adult mosquitoes.

Degradation dynamics and residue determination of pyriclobenzuron in rice and paddy environments

WU Huanqi, WANG Junmin, YANG Kongtan, WANG Xumi, FANG Nan, DUAN Liping, ZHANG Changpeng, WANG Xiangyun

2025, 37(2):  201-208. 

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Objective　To establish a method for determination of pyriclobenzuron (PBU) residues in rice and paddy environments, and to determine the residual amounts and observe the degradation dynamics of PBU.  Methods　In July 2022, the paddies of Zhejiang Academy of Agricultural Sciences were selected as experimental fields, and were divided into the blank control group (no pesticide application), the 1⁃fold⁃concentration pesticide group (1 kg/667 m2), and the 5⁃fold⁃concentration pesticide group (5 kg/667 m2), with a 100 m2 area in each group. At the early tillering stage of rice, 20% suspension of PBU sulfate was sprayed once in the 1⁃fold⁃concentration and 5⁃fold⁃concentration pesticide groups, and rice plants, paddy water and soil samples were collected 2 h, and 1, 2, 3, 5, 7, 11, 14, 21, 28, 35, 49 d and 63 d following spraying PBU, while rice straw, field soil, brown rice and rice husk samples were collected 98 d following spraying. PBU was extracted and purified in samples using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) pretreatment technique, and the PBU contents were determined in samples using ultra⁃high performance liquid chromatography tandem mass spectrometry (UPLC⁃MS/MS). The solvent standard working solution and matrix standard working solution were prepared. A linear regression equation was fitted between PBU concentration (x⁃axis) and peak area (y⁃axis), and the ratio of the slope (k) of the matrix standard curve to the slope (K) of the solvent standard curve was calculated to evaluate the matrix effect of PBU in samples. According to the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), the addition levels of PBU were set at 0.005, 0.050, 5.000, 1 000.000 mg/kg in rice plants, 0.005, 0.050, 2.000, 10.000 mg/kg in paddy water, 0.005, 0.050, 2.000 mg/kg in soil, and 0.005, 0.050, 5.000 mg/kg in brown rice and rice husks. The recovery and relative standard deviation (RSD) of PBU addition were calculated to evaluate the effectiveness of UPLC⁃MS/MS for determination of PBU contents. The first⁃order kinetic equation of PBU concentration was fitted in samples at different sampling time points to analyze the trends in PBU degradation in rice plants, paddy water, and soil, and the half⁃life of PBU was calculated in different samples. Results　There was a good linear relationship between the mass concentration and peak area of PBU at concentrations of 0.000 1 to 0.020 0 mg/kg under solvent and matrix conditions (R2 = 0.985 8 to 0.999 7, t = -0.47 to 1.62, all P values < 0.01). The matrix effects of PBU were 70.26%, 65.42% and 65.12% in rice plants, brown rice and rice husks, indicating a matrix⁃inhibitory effect, and the matrix effect was 87.06% in soils, indicating a weak matrix effect. The recovery of PBU addition was 77.61% to 100.12% in different samples, with RSD of 1.43% to 6.74%, and a limit of quantification (LOQ) of 0.005 mg/kg, and the addition recovery and RSD met the requirements of the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), validating the effectiveness of UPLC⁃MS/MS assay. Following spraying PBU at a dose of 1 kg/667 m2, the half⁃life of PBU was 6.24 d in rice plants and 3.43 d in paddy water samples, respectively. The final residues of PBU were lower than the LOQ of 0.005 mg/kg in brown rice and rice husk samples 98 d following spraying PBU. Following spraying PBU at a dose of 5 kg/667 m2, the half⁃life of PBU was 15.75 d in rice plants and 7.62 d in paddy water samples, respectively. The final residue of PBU was lower than the LOQ of 0.005 mg/kg in brown rice 98 d following spraying PBU, and the final residue of PBU was 0.049 mg/kg in rice husks.  Conclusions　A simple, and highly accurate and precise UPLC⁃MS/MS assay has been developed for determination of PBU residues in rice plants and paddy environments through extraction and purification of PBU from matrix samples using QuEChERS pretreatment. After spraying PBU in paddies, the concentration of PBU gradually decreases in rice plants and paddy water over time, and the final residual concentration is low.

Research progress on techniques for detection of tick⁃borne encephalitis virus infections

LIU Zhuofan, XIE Hao, SUN Xiaoliang, XIA Tao, GUO Junhui

2025, 37(2):  209-216. 

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Tick⁃borne encephalitis is a central nervous system disease caused by infections with tick⁃borne pathogens, which is characterized by severe clinical symptoms, multiple sequelae, and a high fatality rate. Currently, there is no cure for tick⁃borne encephalitis. Tick⁃borne encephalitis virus (TBEV) is the most common pathogen of tick⁃borne encephalitis. Therefore, rapid and accurate detection of TBEV contributes to reducing the mortality of tick⁃borne encephalitis, improving patients' prognosis, and reducing the risk of TBEV transmission. The currently available serological tests for detection of TBEV infections mainly include neutralization test, enzyme⁃linked immunosorbent assay (ELISA), immunofluorescence assay, and nucleic acid tests mainly include polymerase chain reaction (PCR), loop⁃mediated isothermal amplification (LAMP), reverse transcription polymerase spiral reaction, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR⁃associated proteins (Cas)⁃based assays. This review summarizes the progress of researches on serological and nucleic acid tests for detection of TBEV infections, so as to provide insights into prevention and control of tick⁃borne encephalitis.

Current status of anticoccidial drug resistance in China

BAI Yidan, LI Wenting, LUO Wanxin, YU Yuxin, LI Dongfang, ZHAO Junlong, HE Lan

2025, 37(2):  217-222. 

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Avian coccidiosis, an acute parasitic disease that mainly harms chicks, is widely prevalent across the world, which poses a serious threat to poultry industry. Because of the single prophylactic formulations, veterinary clinical treatment of coccidiosis mainly relies on chemically synthesized agents, polyether ionophores and Chinese herbal medicines. The introduction of novel anticoccidial drugs is slow for a long period of time, and there is an increasing problem of anticoccidial drug resistance following long⁃term use, which has become an urgent problem to be solved in poultry industry. This review summarizes the levels of anticoccidial drug resistance across China from 2018 to 2023, and analyzes the resistance to various anticoccidial agents in coccidia. It is indicated that the overall prevalence of anticoccidial drug resistance is high in coccidia, and development of novel anticoccidial agents and products with reduced antibiotics use and alternatives of antibiotics is of an urgent need.