Junfeng Gao

Freshwater fish is widely consumed, yet raw or undercooked freshwater fish can transmit foodborne zoonotic pathogens. Clonorchiasis, caused by Clonorchis sinensis, is a major foodborne parasitic disease associated with hepatobiliary injury and fibrosis. The biological functions and mechanisms underlying liver injury mediated by C. sinensis-derived extracellular vesicles (EVs) encapsulated microRNAs in fibrogenic remodeling remain poorly defined. Here, we isolated and characterized EVs from adult C. sinensis, confirmed their uptake by hepatic stellate cells (HSCs), and identified Csi-bantam as a CsEV-enriched microRNA. Csi-bantam enhances HSC activation and upregulates fibrogenic markers, including α-SMA, COL1A1, and COL3A1, both in vitro and in a mice infection model. Bioinformatic prediction and dual-luciferase assays validated TRIM35 as a target of Csi-bantam. TRIM35 silencing phenocopied Csi-bantam overexpression and increased PI3K/AKT phosphorylation, whereas Csi-bantam inhibition restored TRIM35 and attenuated fibrogenic markers. Collectively, CsEV-associated Csi-bantam promotes HSC activation via the TRIM35-PI3K/AKT axis, suggesting a potential target for antifibrotic intervention in clonorchiasis. These findings provide mechanistic insight into how infection acquired from raw or undercooked freshwater fish harboring infective C. sinensis metacercariae can lead to hepatobiliary injury and fibrogenic remodeling, supporting food safety risk assessment and future prevention of fish-borne parasitic hazards.

Fasciolosis is a globally prevalent trematode infection of major veterinary and public-health relevance. Juveniles migrate through liver tissue for weeks before patency, so clinically important infection may occur while faecal egg output is undetectable, complicating control and interpretation of apparent treatment failure. Triclabendazole (TCBZ) remains central because it targets both immature and adult flukes, but sustained use has been accompanied by geographically expanding reports of reduced efficacy and confirmed resistance. Most alternative fasciolicides, such as albendazole, closantel, oxyclozanide, rafoxanide, clorsulon and nitroxynil, are largely adulticidal and used alone or in combinations, yet reports of reduced efficacy/resistance are increasing worldwide. This review summarises drugs in current use and reported resistance status, and outlines a practical pathway for detecting and confirming resistance. We then appraise leading mechanistic hypotheses for TCBZ resistance as a central case study, organised around microtubule-associated phenotypes, reduced effective drug exposure, genetic architecture with tissue context, stress response and detoxification capacity, and we highlight mechanistic gaps for other fasciolicides. Finally, we discuss management implications, including monitoring-guided stewardship, stage-appropriate drug selection, rational combinations, integrated parasite management, and identify near-term priorities for harmonised surveillance, improved diagnostics and tool development. This review updates the resistance landscape and supports practical, monitoring-guided control of fasciolosis.

Clonorchis sinensis is an important zoonotic parasite that is mainly prevalent in China, Korea, Vietnam and the Russian Far East. To explore the genetic variation and population structure of C. sinensis, an in silico analysis was conducted based on mitochondrial cytochrome c oxidase subunit 1 (COX1), ribosomal internal transcribed spacer 1 (ITS1) and ribosomal internal transcribed spacer 2 (ITS2) sequences. The sequences obtained from NCBI were truncated for further analyses, including haplotype network, phylogenetic, gene flow, diversity and neutrality analyses. The results showed that there were 20, 11 and 4 haplotypes for COX1, ITS1 and ITS2, respectively. The results of both the haplotype network and phylogenetic analyses indicated that the haplotypes for each type of sequence from the same country were not all clustered together. Haplotype diversity values were all lower than 0.5. Values of nucleotide diversity were higher than 0.005, except for ITS2. Tajima’s D and Fu’s Fs values were all negative, and p-values showed significant differences, indicating that the population of C. sinensis is growing. Fst values were all lower than 0.05. In conclusion, this study found that there are specific variations of C. sinensis in different countries, and the population of this parasite is growing with less genetic variation. The findings provide a crucial foundation for understanding the molecular epidemiology and population dynamics of C. sinensis.

Crytosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are important diarrheal pathogens with a global distribution that threatens the health of humans and animals. Despite cattle being potential transmission hosts of these protozoans, the associated risks to public health have been neglected. In the present study, a total of 1155 cattle fecal samples were collected from 13 administrative regions of Heilongjiang Province. The prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 5.5% (64/1155; 95% CI: 4.2–6.9), 3.8% (44/1155; 95% CI: 2.7–4.9), and 6.5% (75/1155; 95% CI: 5.1–7.9), respectively. Among these positive fecal samples, five Cryptosporidium species (C. andersoni, C. bovis, C. ryanae, C. parvum, and C. occultus), two G. duodenalis assemblages (E and A), and eight E. bieneusi genotypes (BEB4, BEB6, BEB8, J, I, CHS7, CHS8, and COS-I) were identified. Phylogenetic analysis showed that all eight genotypes of E. bieneusi identified in the present study belonged to group 2. It is worth noting that some species/genotypes of these intestinal protozoans are zoonotic, suggesting a risk of zoonotic disease transmission in endemic areas. The findings expanded our understanding of the genetic composition and zoonotic potential of Cryptosporidium spp., G. duodenalis, and E. bieneusi in cattle in Heilongjiang Province.

Fasciolosis is a significant zoonotic and common parasitic disease for animals and humans, creating public health concerns worldwide. This study retrieved articles related to the occurrence of Fasciola hepatica and Fasciola gigantica in sheep and goats in China by searching five databases: PubMed, ScienceDirect, the Chinese National Knowledge Infrastructure (CNKI), Wanfang Data, and the VIP Chinese Journal Database. A total of 60 valid articles were captured. The pooled prevalence of ovine and caprine fasciolosis was 26.00%. It was also found to be higher in the subgroups of Northwest China and Shaanxi Province, as well as in areas with a high altitude, rainfall of ≥800 mm, and temperature ranging between 10 °C and 20 °C. Analysis of the type of season and sampling years showed significant (p < 0.05) difference. In other subgroups, sheep (34.74%), hosts aged over 2 years (32.26%), females (48.33%) and free-range animals (26.83%) showed a higher disease prevalence. These results indicated that ovine and caprine fasciolosis was widely distributed, especially in Northwest China. The sampling years and the type of season are risk factors for the prevalence of ovine and caprine fasciolosis. Therefore, strategies for ovine and caprine fasciolosis control should be developed based on these epidemic risk factors, which will reduce the prevalence of fasciolosis in China.

Ticks carry and transmit a variety of pathogens, which are very harmful to humans and animals. To characterize the microbial interactions in ticks, we analysed the microbiota of the hard ticks, Dermacentor silvarum, Ixodes persulcatus, and Haemaphysalis concinna, using 16S rRNA, showing that microbial interactions are underappreciated in terms of shaping arthropod microbiomes. The results show that the bacterial richness and microbiota structures of these three tick species had significant differences. Interestingly, the bacterial richness (Chao1 index) of all ticks decreased significantly after they became engorged. All the operational taxonomic units (OTUs) were assigned to 26 phyla, 67 classes, 159 orders, 279 families, and 627 genera. Microbial interactions in D. silvarum demonstrated more connections than in I. persulcatus and H. concinna. Bacteria with a high abundance were not important families in microbial interactions. Positive interactions of Bacteroidaceae and F_Solibacteraceae Subgroup 3 with other bacterial families were detected in all nine groups of ticks. This study provides an overview of the microbiota structure and interactions of three tick species and improves our understanding of the role of the microbiota in tick physiology and vector capacity, thus being conducive to providing basic data for the prevention of ticks and tick-borne diseases.

Background: Clonorchiasis, an infectious disease caused by the liver fluke Clonorchis sinensis, may lead to the development of liver and gallbladder diseases, and even cholangiocarcinoma (CCA). However, the pathogenesis, host-pathogen interaction, and diagnostic markers for clonorchiasis remain unclear. Methods: Eighteen rabbits were randomly divided into control group (n = 9) and C. sinensis-infected group (n = 9), and their plasma samples were collected at 7, 14, 28, and 63 days post-infection (dpi). Biochemical indices and metabolites in different infection periods were detected. A non-targeted ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach was employed to investigate the metabolic profiles of plasma in rabbits, and related metabolic pathways of differential metabolites and correlation between candidate biochemical indices and differential metabolites were analyzed. Finally, the candidate biomarkers were verified with human samples using a targeted metabolomics method. Results: The result of biochemical indices indicated C. sinensis infection would affect the liver function biochemical indices, especially alanine aminotransferase, aspartate transaminase (AST), glutamyl transpeptidase (GGT), total bile acid, high-density lipoprotein, and cholinesterase. The metabonomic results showed that 58, 212, 23, and 21 differential metabolites were identified in different phases of the infection. Multivariate statistical analysis of differential metabolites revealed distinct metabolic signatures during different phases of infection, with most of these signatures being observed at 14 dpi, which mainly influences the amino acid metabolisms. For metabolites and biochemical indices, AST, GGT, hypoxanthine, l-pipecolic acid, and d-glucuronate represented potential noninvasive biomarkers for the diagnosis of C. sinensis (P < 0.05 and AUC > 0.8). Furthermore, GGT and d-glucuronate levels were positively correlated with the infection (r(28) = 0.98, P < 0.0001) and showed excellent diagnostic performance (AUC = 0.972; 95% confidence interval, 0.921 to 1.000). Conclusions: The present results provide new insights into plasma metabolic changes in rabbits during C. sinensis infection, and the potential biomarker may be used for developing an effective method to diagnose clonorchiasis in the future. Graphical