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Background: Clonorchiasis has significant socioeconomic importance in endemic areas; however, studies investigating the disease burden in specific sub-regions are lacking. This study aims to address the gap by quantifying the current disease burden caused by clonorchiasis in Guangdong province and assessing its distribution characteristics., Methods: Comprehensive measures, including prevalence rates, disability-adjusted life years (DALYs), and direct medical costs, were used to assess the disease burden of clonorchiasis. To estimate the prevalence rate, the number of infections was divided by the examined population, based on the annual surveillance data on clonorchiasis cases during 2016-2021. The calculation of DALYs was based on the epidemiological parameters according to the definition issued by the World Health Organization. Cost data of clonorchiasis were utilized to quantify the direct medical costs. The distribution characteristics of disease burden were assessed through comparisons of groups of population defined by geographic area, time, and characteristics of people., Results: In 2021, clonorchiasis posed a significant disease burden in Guangdong Province. The prevalence rate was found to be 4.25% [95% CI (4.02%, 4.49%)], with an associated burden of DALYs of 406,802.29 [95% CI (329,275.33, 49,215,163.78)] person-years. The per-case direct medical costs of patients with clonorchiasis were estimated to be CNY 7907.2 (SD = 5154.4). Notably, while the prevalence rate and DALYs showed a steady decrease from 2016 to 2020, there was a rising trend in 2021. Spatial clustering of clonorchiasis cases and DALYs was also observed, particularly along the Pearl River and Han River. This suggests a concentration of the disease in these regions. Furthermore, significant differences in prevalence rates were found among various demographic groups, including sex, age, occupation, and education level. Additionally, patients with longer hospital stays were more likely to incur higher direct medical costs., Conclusions: The burden of clonorchiasis in Guangdong Province remains high, despite significant progress achieved through the implementation of the prevention and control programs. It is suggested that measures should be taken based on the distribution characteristics to maximize the effectiveness of prevention and control, with a primary focus on key populations and areas., (© 2024. The Author(s).)

Background: Clonorchiasis remains a serious public health problem. However, the molecular mechanism underlying clonorchiasis remains largely unknown. Amino acid (AA) metabolism plays key roles in protein synthesis and energy sources, and improves immunity in pathological conditions. Therefore, this study aimed to explore the AA profiles of spleen in clonorchiasis and speculate the interaction between the host and parasite. Methods: Here targeted ultrahigh performance liquid chromatography multiple reaction monitoring mass spectrometry was applied to discover the AA profiles in spleen of rats infected with Clonorchis sinensis . Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis (KEGG) was performed to characterize the dysregulated metabolic pathways. Results: Pathway analysis revealed that phenylalanine, tyrosine, and tryptophan biosynthesis and β-alanine metabolism were significantly altered in clonorchiasis. There were no significant correlations between 14 significant differential AAs and interleukin (IL)-1β. Although arginine, asparagine, histidine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine were positively correlated with IL-6, IL-10, tumor necrosis factor (TNF)-α as well as aspartate aminotransferase and alanine aminotransferase; β-alanine and 4-hydroxyproline were negatively correlated with IL-6, IL-10, and TNF-α. Conclusion: This study reveals the dysregulation of AA metabolism in clonorchiasis and provides a useful insight of metabolic mechanisms at the molecular level.

Clonorchiasis is a zoonotic disease mainly caused by eating raw fish and shrimp, and there is no vaccine to prevent it. More than 30 million people are infected worldwide, of which China alone accounts for about half, and is one of the countries most seriously affected by Clonorchiasis. In this work, we formulate a novel Ordinary Differential Equation (ODE) model to discuss the biological attributes of fish within authentic ecosystems and the complex lifecycle of Clonorchis sinensis. This model includes larval fish, adult fish, infected fish, humans, and cercariae. We derive the basic reproduction number and perform a rigorous stability analysis of the proposed model. Numerically, we use data from 2016 to 2021 in Guangxi, China, to discuss outbreaks of Clonorchiasis and obtain the basic reproduction number R 0 =1.4764. The fitted curve appropriately reflects the overall trend and replicates a low peak in the case number of Clonorchiasis. By reducing the release rate of cercariae in 2018, the fitted values of Clonorchiasis cases dropped rapidly and almost disappeared. If we decrease the transmission rate from infected fish to humans, Clonorchiasis can be controlled. Our studies also suggest that strengthening publicity education and cleaning water quality can effectively control the transmission of Clonorchiasis in Guangxi, China., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objective: Foodborne zoonotic trematodes (FZT), which infect the liver, lungs and intestines of humans, are an emerging public health concern in tropical countries including Vietnam. In northern Vietnam, Clonorchis sinensis is recognised as the most important species of the FZT. Up to now, small-scale studies conducted in the country have indicated that health education could improve participants' knowledge and practices related to clonorchiasis, however strong evidence is still lacking. We conducted an intervention trial between 2020 and 2021 in four communes in Yen Bai and Thanh Hoa province, aiming to evaluate the impact of an extended educational package on knowledge, attitude and practices related to clonorchiasis, as well as on knowledge on the large liver fluke and minute intestinal flukes., Methods: To this end, baseline and post-intervention questionnaires were conducted. Generalised estimating equations models were run to analyse the impact of the intervention on knowledge, attitudes and practices over time in the intervention versus control group. Finally, 172 people in the intervention group and 162 in the control group were included for analysis., Results: Results demonstrated that the intervention significantly improved clonorchiasis knowledge, attitudes and practices, with higher odds for a correct knowledge response (odds ratio (OR) = 2.80, 95% confidence interval (CI) = 1.84-4.27, p-value < 0.001), increased average mean attitude score (mean = 0.363, 95%CI = 0.182-0.544, p-value < 0.001), and a reduced odds of consuming raw fish (OR = 0.15, 95%CI = 0.06-0.40, p = 0.002) in the intervention group compared to the control group post-intervention versus baseline. Additionally, participants in the intervention group exhibited enhanced basic knowledge of the large liver fluke and minute intestinal flukes post-intervention., Conclusions: These findings demonstrate the potential of health education programs in preventing clonorchiasis in endemic areas, emphasising the importance of continued health education as a critical component of integrated control programs for clonorchiasis., (© 2024 The Authors Tropical Medicine & International Health Published by John Wiley & Sons Ltd.)

Clonorchis sinensis , Opisthorchis viverrini , and Opisthorchis felineus are important liver flukes that cause a considerable public health burden in eastern Asia, southeastern Asia, and eastern Europe, respectively. The life cycles are complex, involving humans, animal reservoirs, and two kinds of intermediate hosts. An interplay of biological, cultural, ecological, economic, and social factors drives transmission. Chronic infections are associated with liver and biliary complications, most importantly cholangiocarcinoma. With regard to diagnosis, stool microscopy is widely used in epidemiologic surveys and for individual diagnosis. Immunologic techniques are employed for screening purposes, and molecular techniques facilitate species differentiation in reference laboratories. The mainstay of control is preventive chemotherapy with praziquantel, usually combined with behavioral change through information, education and communication, and environmental control. Tribendimidine, a drug registered in the People's Republic of China for soil-transmitted helminth infections, shows potential against both C. sinensis and O. viverrini and, hence, warrants further clinical development. Novel control approaches include fish vaccine and biological control. Considerable advances have been made using multi-omics which may trigger the development of new interventions. Pressing research needs include mapping the current distribution, disentangling the transmission, accurately estimating the disease burden, and developing new diagnostic and treatment tools, which would aid to optimize control and elimination measures., Competing Interests: The authors declare no conflict of interest.

Background: Clonorchiasis and opisthorchiasis, caused by the liver flukes Clonorchis sinensis and Opisthorchis viverrini respectively, represent significant neglected tropical diseases (NTDs) in Asia. The co-existence of these pathogens in overlapping regions complicates effective disease control strategies. This study aimed to clarify the distribution and interaction of these diseases within Southeast Asia., Methods: We systematically collated occurrence records of human clonorchiasis (n = 1809) and opisthorchiasis (n = 731) across the Southeast Asia countries. Utilizing species distribution models incorporating environmental and climatic data, coupled machine learning algorithms with boosted regression trees, we predicted and distinguished endemic areas for each fluke species. Machine learning techniques, including geospatial analysis, were employed to delineate the boundaries between these flukes., Results: Our analysis revealed that the endemic range of C. sinensis and O. viverrini in Southeast Asia primarily spans across part of China, Vietnam, Thailand, Laos, and Cambodia. During the period from 2000 to 2018, we identified C. sinensis infections in 84 distinct locations, predominantly in southern China (Guangxi Zhuang Autonomous Region) and northern Vietnam. In a stark contrast, O. viverrini was more widely distributed, with infections documented in 721 locations across Thailand, Laos, Cambodia, and Vietnam. Critical environmental determinants were quantitatively analyzed, revealing annual mean temperatures ranging between 14 and 20 °C in clonorchiasis-endemic areas and 24-30 °C in opisthorchiasis regions (P < 0.05). The machine learning model effectively mapped a distinct demarcation zone, demonstrating a clear separation between the endemic areas of these two liver flukes with AUC from 0.9 to1. The study in Vietnam delineates the coexistence and geographical boundaries of C. sinensis and O. viverrini, revealing distinct endemic zones and a transitional area where both liver fluke species overlap., Conclusions: Our findings highlight the critical role of specific climatic and environmental factors in influencing the geographical distribution of C. sinensis and O. viverrini. This spatial delineation offers valuable insights for integrated surveillance and control strategies, particularly in regions with sympatric transmission. The results underscore the need for tailored interventions, considering regional epidemiological variations. Future collaborations integrating eco-epidemiology, molecular epidemiology, and parasitology are essential to further elucidate the complex interplay of liver fluke distributions in Asia., (© 2024. The Author(s).)

Background: Multi-factorial reasons are an induction to cause cancer. Different infections and infestations with viruses, bacteria, and parasites have been detected for many years to be related to human carcinogenesis., Purpose: The study aimed to review all ideas of tumor carcinogenesis and its associations with parasitic infections and infestations., Methods: We reviewed several articles (published and imprinted) by selecting, extracting, and synthesizing data about the relationship between cancers and parasites., Results: Several helminths infections as schistosomiasis, are highly carcinogenic agents for bladder cancer, whereas trypanosomiasis has a bi-model role in cancer development. Leishmaniasis may be a cause of hepatocarcinoma, skin cancer, and lymphomas. In addition, malaria appears to be causative in the carcinogenesis of some cancers; as Burkitt lymphoma. Also, data from previous studies suggested that Strongyloides stercoralis may be a relevant co-factor in lymphomas., Conclusion: There are different mechanisms of parasitic infection to be enhancing in carcinogenesis of cancer in human., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Background: The latest national survey on the distribution of human parasites in China demonstrated that Guangdong was among the endemic provinces with the highest Clonorchis sinensis infection rates. High-resolution, age- and gender-specific risk maps of the temporal and spatial distributions are essential for the targeted control work., Methods: Disease data on the prevalence of C. sinensis infection from 1990 onwards, either age- and gender-specific or aggregated across age and gender, were collected through systematic review and four large-scale surveys in Guangdong Province. Environmental and socioeconomic variables were obtained from open-access databases and employed as potential predictors. A Bayesian geostatistical model was developed to estimate the C. sinensis infection risk at high spatial resolution., Results: The final dataset included 606 surveys at 463 unique locations for C. sinensis infection. Our findings suggested that following an initial increase and stabilization, the overall population-adjusted prevalence had declined to 2.2% (95% Bayesian credible interval: 1.7-3.0%) in the period of 2015 onwards. From 2015 onwards, moderate and high infection risks were found in the northern regions (e.g. Heyuan and Shaoguan cities) and the southern Pearl River Delta (e.g. Foshan, Zhongshan, Zhuhai and Jiangmen cities), respectively. Age- and gender-specific risk maps revealed that males had a higher infection risk than females, and the infection risk was higher in adults compared to children., Conclusions: Our high-resolution risk maps of C. sinensis infection in Guangdong Province identified the spatial, temporal, age and gender heterogeneities, which can provide useful information assisting tailored control strategies., (© 2024. The Author(s).)

To investigate the effects of co-infection with Clonorchis sinensis (C. sinensis) on T cell exhaustion levels in patients with chronic hepatitis B, we enrolled clinical cases in this study, including the patients with concomitant C. sinensis and HBV infection. In this study, we detected inhibitory receptors and cytokine expression in circulating CD4+ and CD8+ T cells by flow cytometry. PD-1 and TIM-3 expression levels were significantly higher on CD4+ T and CD8+ T cells from co-infected patients than on those from the HBV patients. In addition, CD4+ T cells and CD8+ T cells function were significantly inhibited by C. sinensis and HBV co-infection compared with HBV single infection, secreting lower levels of Interferon gamma (IFN-γ), Interleukin-2 (IL-2), and TNF-α. Our current results suggested that C. sinensis co-infection could exacerbate T cell exhaustion in patients with chronic hepatitis B. PD-1 and TIM-3 could be novel biomarkers for T cell exhaustion in patients with Clonorchis sinensis and chronic hepatitis B co-infection. Furthermore, it may be one possible reason for the weaker response to antiviral therapies and the chronicity of HBV infection in co-infected patients. We must realize the importance of C. sinensis treatment for HBV-infected patients. It might provide useful information for clinical doctors to choose the right treatment plans.

Objective: To evaluate the effect of information-knowledge-attitude-practice (IKAP) theory-based health education interventions on clonorchiasis control among community residents and primary school students in Torch High-tech Development Zone of Zhongshan City, so as to provide insights into formulation of clonorchiasis control strategies among different types of populations., Methods: Residents were randomly sampled from two communities and students from two primary schools using a random cluster sampling method in Torch High-tech Development Zone, Zhongshan City from 2019 to 2021, and health education pertaining to clonorchiasis control was performed based on the IKAP theory. The changes in the awareness of basic clonorchiasis control knowledge and belief, self-reported risk of Clonorchis sinensis infections and dietary behaviors were compared among community residents and primary school students before and after health education interventions., Results: The participants included 146 male and 151 female community residents and 158 male and 153 female primary school students, with no significant difference detected in gender distribution (χ 2 = 0.16, P > 0.05). The mean awareness of basic clonorchiasis control knowledge increased by 44.71% and 43.28% among primary school students and community residents 6 months following health education, and there were significant differences in the awareness of each item of basic clonorchiasis control knowledge before and after health education (χ 2 = 41.53 to 284.44, all P values < 0.05). The proportions of primary school students and community residents thinking very high and high risks of C. sinensis infections increased from 9.35% and 6.71% before health education to 22.15% and 37.75% after health education, but only the difference of the attitudes in community residents achieve statistical significance (χ 2 = 92.18, P < 0.05). The frequency of separation of raw and cooked foods with chopping board and knife significantly increased among community residents and primary school students following health education (χ 2 = 16.04 to 62.65, all P values < 0.05). The frequency of eating raw freshwater fish (χ 2 = 32.85, P < 0.05), fish congee (χ 2 = 7.02, P < 0.05) and fish fillet hot pot (χ 2 = 4.88, P < 0.05) significantly reduced among primary school students following health education, while only the frequency and proportions of eating raw freshwater fish have significantly reduced in community residents (χ 2 = 11.77, P < 0.05)., Conclusions: Health education interventions based on the IKAP theory are effective to increase the awareness of clonorchiasis control knowledge and improve dietary behaviors associated with C. sinensis infections among community residents and primary school students in Zhongshan City.

Background: Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis), is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma. It has been well known that NO from chronic inflammation responses are thought to be a major component of the damage and ultimate carcinogenesis ESPs such as nitric oxide synthase interacting protein (NOSIP) are thought to enhance the damage. The objective of this study was to identify the protein candidates interact with recombinant CsNOSIP (rCsNOSIP) and explore their role involved in CCA development or progression., Methods: We applied HuProt microarray containing 21,000 probe sets for a systematic identification of rCsNOSIP-binding proteins and grouped binding hits by gene function. Pull-down assays were used to confirm the interaction of rCsNOSIP with alveolar soft part sarcoma (ASPSCR-1) and sirtuins 5 (Sirt-5). ASPSCR-1/Sirt-5 over-expression and siRNA knockdown experiments were employed for obtain of ASPSCR-1/Sirt-5 high or low expression (ASP-oe/Sirt5-oe or ASP-si/Sirt5-si) cholangiocarcinoma cell line (CCLP-1) cells. Nitric oxide (NO) and reactive oxygen species assay (ROS) as well as cell proliferation and wound-healing assays were performed to observe the effect of rCsNOSIP on ASP-oe/Sirt5-oe or ASP-si/Sirt5-si CCLP-1 cells., Results: Seventy candidate proteins protein "hits" were detected as rCsNOSIP-binding proteins by HuProt microarray and bioinformatics analysis. Pull down assay showed that ASPSCR-1 and Sirt-5 could interact with rCsNOSIP. In addition, endotoxin-free-rCsNOSIP could increase the production of NO and ROS and promote the migration of CCLP-1 cells, while its effect on enhancing cell proliferation was not significant. Furthermore, ROS/NO production, proliferation, or migration were increased in ASP-si or Sirt5-si CCLP-1 cells but decreased in Asp-oe or Sirt5-oe CCLP-1 cells when stimulated with rCsNOSIP., Conclusions: Our findings suggest that CsNOSIP as a component of CsESPs might promote the development and invasion of CCA and Sirt5/ ASPSCR1 as host molecules might play a novel protective role against adverse stimulus during C. sinensis infection. This work supports the idea that CsESPs induce the occurrence and progression of CCA through ROS/RNS-induced oxidative and nitrative DNA damage., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Bian et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Clonorchiasis remains a non-negligible global zoonosis, causing serious socioeconomic burdens in endemic areas. Clonorchis sinensis infection typically elicits Th1/Th2 mixed immune responses during the course of biliary injury and periductal fibrosis. However, the molecular mechanism by which C. sinensis juvenile initially infects the host remains poorly understood., Methods: The BALB/c mouse model was established to study early infection (within 7 days) with C. sinensis juveniles. Liver pathology staining and observation as well as determination of biochemical enzymes, blood routine and cytokines in blood were conducted. Furthermore, analysis of liver transcriptome, proteome and metabolome changes was performed using multi-omics techniques. Statistical analyses were performed using Student's t-test., Results: Histopathological analysis revealed that liver injury, characterized by collagen deposition and inflammatory cell infiltration, occurred as early as 24 h of infection. Blood indicators including ALT, AST, WBC, CRP and IL-6 indicated that both liver injury and systemic inflammation worsened as the infection progressed. Proteomic data showed that apoptosis and junction-related pathways were enriched within 3 days of infection, indicating the occurrence of liver injury. Furthermore, proteomic and transcriptomic analysis jointly verified that the detoxification and antioxidant defense system was activated by enrichment of glutathione metabolism and cytochrome P450-related pathways in response to acute liver injury. Proteomic-based GO analysis demonstrated that biological processes such as cell deformation, proliferation, migration and wound healing occurred in the liver during the early infection. Correspondingly, transcriptomic results showed significant enrichment of cell cycle pathway on day 3 and 7. In addition, the KEGG analysis of multi-omics data demonstrated that numerous pathways related to immunity, inflammation, tumorigenesis and metabolism were enriched in the liver. Besides, metabolomic screening identified several metabolites that could promote inflammation and hepatobiliary periductal fibrosis, such as CA7S., Conclusions: This study revealed that acute inflammatory injury was rapidly triggered by initial infection by C. sinensis juveniles in the host, accompanied by the enrichment of detoxification, inflammation, fibrosis, tumor and metabolism-related pathways in the liver, which provides a new perspective for the early intervention and therapy of clonorchiasis., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Clonorchis sinensis is a carcinogenic liver fluke that causes clonorchiasis-a neglected tropical disease (NTD) affecting ~35 million people worldwide. No vaccine is available, and chemotherapy relies on one anthelmintic, praziquantel. This parasite has a complex life history and is known to infect a range of species of intermediate (freshwater snails and fish) and definitive (piscivorous) hosts. Despite this biological complexity and the impact of this biocarcinogenic pathogen, there has been no previous study of molecular variation in this parasite on a genome-wide scale. Here, we conducted the first extensive nuclear genomic exploration of C. sinensis individuals (n = 152) representing five distinct populations from mainland China, and one from Far East Russia, and revealed marked genetic variation within this species between "northern" and "southern" geographical regions. The discovery of this variation indicates the existence of biologically distinct variants within C. sinensis, which may have distinct epidemiology, pathogenicity and/or chemotherapic responsiveness. The detection of high heterozygosity within C. sinensis specimens suggests that this parasite has developed mechanisms to readily adapt to changing environments and/or host species during its life history/evolution. From an applied perspective, the identification of invariable genes could assist in finding new intervention targets in this parasite, given the major clinical relevance of clonorchiasis. From a technical perspective, the genomic-informatic workflow established herein will be readily applicable to a wide range of other parasites that cause NTDs., (© 2023 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.)

Background: Foodborne trematodiases (FBTs) are a group of trematodes targeted for control as part of the World Health Organization (WHO) road map for neglected tropical diseases from 2021 to 2030. Disease mapping; surveillance; and capacity, awareness, and advocacy building are critical to reach the 2030 targets. This review aims to synthesise available data on FBT prevalence, risk factors, prevention, testing, and treatment., Methods: We searched the scientific literature and extracted prevalence data as well as qualitative data on the geographical and sociocultural risk factors associated with infection, preventive/protective factors, and methods and challenges of diagnostics and treatment. We also extracted WHO Global Health Observatory data representing the countries that reported FBTs during 2010 to 2019., Results: One hundred and fifteen studies reporting data on any of the 4 FBTs of focus (Fasciola spp., Paragonimus spp., Clonorchis sp., and Opisthorchis spp.) were included in the final selection. Opisthorchiasis was the most commonly reported and researched FBT, with recorded study prevalence ranging from 0.66% to 88.7% in Asia, and this was the highest FBT prevalence overall. The highest recorded study prevalence for clonorchiasis was 59.6%, reported in Asia. Fascioliasis was reported in all regions, with the highest prevalence of 24.77% reported in the Americas. The least data was available on paragonimiasis, with the highest reported study prevalence of 14.9% in Africa. WHO Global Health Observatory data indicated 93/224 (42%) countries reported at least 1 FBT and 26 countries are likely co-endemic to 2 or more FBTs. However, only 3 countries had conducted prevalence estimates for multiple FBTs in the published literature between 2010 to 2020. Despite differing epidemiology, there were overlapping risk factors for all FBTs in all geographical areas, including proximity to rural and agricultural environments; consumption of raw contaminated food; and limited water, hygiene, and sanitation. Mass drug administration and increased awareness and health education were commonly reported preventive factors for all FBTs. FBTs were primarily diagnosed using faecal parasitological testing. Triclabendazole was the most reported treatment for fascioliasis, while praziquantel was the primary treatment for paragonimiasis, clonorchiasis, and opisthorchiasis. Low sensitivity of diagnostic tests as well as reinfection due to continued high-risk food consumption habits were common factors., Conclusion: This review presents an up-to-date synthesis on the quantitative and qualitative evidence available for the 4 FBTs. The data show a large gap between what is being estimated and what is being reported. Although progress has been made with control programmes in several endemic areas, sustained effort is needed to improve surveillance data on FBTs and identify endemic and high-risk areas for environmental exposures, through a One Health approach, to achieve the 2030 goals of FBT prevention., Competing Interests: None declared., (Copyright: © 2023 Tidman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Clonorchiasis is an important foodborne parasitic disease. The omics-based-techniques could illuminate parasite biology and further make innovations in the research for parasitic diseases. However, knowledge about the serum metabolic profiles and related metabolic pathways in clonorchiasis is very limited., Methods: A untargeted ultra-high performance liquid tandem chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) was used to profile the serum metabolites of rats at both 4 and 8 weeks post infection (wpi) with Clonorchis sinensis ( C. sinensis ). Additionally, multivariate statistical analysis methods were employed to identify differential metabolites. Next, serum amino acids and phosphatidylcholines (PCs) levels were determined by targeted metabolomics analysis., Result: A total of 10530 and 6560 ions were identified in ESI+ and ESI- modes. The levels of phosphatidylcholines, glycerophosphocholine and choline were significantly changed, with the shift in lipid metabolism. Significant changes were also observed in amino acids (isoleucine, valine, leucine, threonine, glutamate and glutamine). Targeted analysis showed that BCAAs (isoleucine, valine, leucine) levels significantly increased at 4 wpi and decreased at 8 wpi; threonine was increased at 8 wpi, whereas glutamate and glutamine showed a decreasing trend at 8 wpi. Additionally, the level of 17 PCs were significantly changed in infected rats. Marked metabolic pathways were involved in clonorchiasis, including glycerophospholipid metabolism, alanine, aspartate and glutamate metabolism, histidine metabolism and pyrimidine metabolism., Conclusion: These results show that C. sinensis infection can cause significant changes in the rat serum metabolism, especially in amino acids and lipids. The metabolic signature together with perturbations in metabolic pathways could provide more in depth understanding of clonorchiasis and further make potential therapeutic interventions., Competing Interests: The authors declared that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Han, Zhang, Ding, Li, Zhang, Jiang, Duan, Sun, Hu and Gao.)

Chronic human liver fluke infections caused by Opisthorchis viverrini and Clonorchis sinensis can last for decades and cause liver and biliary diseases, including life-threatening pathology prior to cholangiocarcinoma (CCA). CCA generally has a poor prognosis. Serological diagnosis can support parasitological examination in diagnosing disease and screening for the risk of CCA. Here, we present an improved and innovative lateral flow immunochromatographic test (ICT) kit that uses whole-blood samples (WBS) rather than serum to diagnose human opisthorchiasis, which also successfully diagnosed human clonorchiasis. This ICT includes a soluble worm extract of O . viverrini 347). Simulated WBS were obtained by the spiking infection sera with red blood cells. The diagnostic sensitivity, specificity, positive and negative predictive values, and accuracy for detecting opisthorchiasis were 95.5%, 87.0%, 80.5%, 97.2%, and 90.1%, respectively. For clonorchiasis, these findings were 85.7%, 87.0%, 53.6%, 97.2%, and 86.8%, respectively. Combined for both diseases, they were 93.2%, 87.0%, 84.0%, 94.6%, and 89.6%, respectively. The ICT kit can possibly replace the ICT platforms for antibody detection in serum samples in field surveys in remote areas where sophisticated equipment is not available.n = 347). Simulated WBS were obtained by the spiking infection sera with red blood cells. The diagnostic sensitivity, specificity, positive and negative predictive values, and accuracy for detecting opisthorchiasis were 95.5%, 87.0%, 80.5%, 97.2%, and 90.1%, respectively. For clonorchiasis, these findings were 85.7%, 87.0%, 53.6%, 97.2%, and 86.8%, respectively. Combined for both diseases, they were 93.2%, 87.0%, 84.0%, 94.6%, and 89.6%, respectively. The ICT kit can possibly replace the ICT platforms for antibody detection in serum samples in field surveys in remote areas where sophisticated equipment is not available.

Clonorchis sinensis is a carcinogenic liver fluke that causes clonorchiasis in humans. Clonorchiasis is prevalent in East Asian countries, and approximately 15–20 million individuals are estimated to be infected with this fluke globally. This review highlights the current status of C. sinensis and clonorchiasis in Korea from the epidemiological perspective involving the analysis of humans and intermediate hosts. Despite the recent decline in C. sinensis infection rate in Korea, C. sinensis infections remain endemic in 5 major river basins (Han-gang, Geum-gang, Seomjin-gang, Yeongsan-gang and Nakdong-gang; gang means river) with a high incidence of cholangiocarcinoma. A noticeable pattern involves increasing mild infections among patients diagnosed positive for C. sinensis eggs. The infection rate of C. sinensis metacercariae in the second intermediate host, freshwater fish, is also maintained at a substantial level. Thus, the One Health approach integrating different sectors and disciplines is recommended to accelerate and sustain control of C. sinensis , thereby leading to successful eradication. Health promotion via information dissemination and health education should be extended to prevent the consumption of raw freshwater fish by residents living in high-risk areas.

The foodborne liver trematode Opisthorchis felineus (Rivolta, 1884) is a member of the triad of phylogenetically related epidemiologically important Opisthorchiidae trematodes, which also includes O. viverrini (Poirier, 1886) and Clonorchis sinensi s (Loos, 1907). Despite similarity in the life cycle, Opisthorchiidae liver flukes also have marked differences. Two species ( O. viverrini and C. sinensis ) are recognized as Group 1A biological carcinogens, whereas O. felineus belongs to Group 3A. In this review, we focus on these questions: Are there actual differences in carcinogenicity among these 3 liver fluke species? Is there an explanation for these differences? We provide a recent update of our knowledge on the liver fluke O. felineus and highlight its differences from O. viverrini and C. sinensis . In particular, we concentrate on differences in the climate of endemic areas, characteristics of the life cycle, the range of intermediate hosts, genomic and transcriptomic features of the pathogens, and clinical symptoms and morbidity of the infections in humans. The discussion of these questions can stimulate new developments in comparative studies on the pathogenicity of liver flukes and should help to identify species-specific features of opisthorchiasis and clonorchiasis pathogenesis.

This study provides an overview of the current status of clonorchiasis and cholangiocarcinoma (CCA), and their relationship in Korea during 2012-2020. Data were obtained from the Health Insurance Review & Assessment Service of Korea. Cluster, trend, and correlation analyses were performed. Gyeongsangnam-do and Seoul had the highest average number of cases (1,026 and 4,208) and adjusted rate (306 and 424) for clonorchiasis and CCA, respectively. The most likely clusters (MLC) for clonorchiasis and CCA were Busan/Gyeongsangnam-do/Ulsan/Daegu/Gyeongsangbuk-do (Relative Risk; RR = 4.55, Likelihood Ratio; LLR = 9,131.115) joint cluster and Seoul (RR = 2.29, LLR = 7,602.472), respectively. The MLC for clonorchiasis was in the southeastern part of Korea, while that for CCA was in the southern part. Clonorchiasis showed a decreasing trend in the southeastern districts, while increased in the southwestern districts. Cities in the central region had a decreasing trend, while the western districts had an increasing trend. In most adults (30-59), infection rate of clonorchiasis showed a significant decrease until 2018, while thereafter increased, although not significant. CCA showed a sharply decreasing tendency. The incidence of clonorchiasis and CCA were positively correlated. In general, the correlation was weak (r = 0.39, P < 0.001), but it was strongly positive around the 4 river basins (r = 0.74, P < 0.001). This study might provide an analytic basis for developing an effective system against clonorchiasis and CCA.

Heliminthic paramyosin is a multifunctional protein that not only acts as a structural protein in muscle layers but as an immune-modulatory molecule interacting with the host immune system. Previously, we found that paramyosin from Clonorchis sinensis (CsPmy) is bound to human complement C9 protein (C9). To analyze the C9 binding region on CsPmy, overlapping recombinant fragments of CsPmy were produced and their binding activity to human C9 was investigated. The fragmental expression of CsPmy and C9 binding assays revealed that the C9 binding region was located at the C-terminus of CsPmy. Further analysis of the C-terminus of CsPmy to narrow the C9 binding region on CsPmy indicated that the region flanking731Leu-780 Leu was a potent C9 binding region. The CsPmy fragments corresponding to the region effectively inhibited human C9 polymerization. These results provide a precise molecular basis for CsPmy as a potent immunomodulator to evade host immune defenses by inhibiting complement attack.

Background: Clonorchiasis is a widespread yet neglected foodborne disease with over 85% of all cases found in China. Guangxi province, located in southeastern China, ranks among the highest endemic provinces. We explore the epidemiological status and determinants of Clonorchis sinensis (C. sinensis) infection in humans and freshwater fish in Guangxi, China., Methods: Data on C. sinensis infection in humans from January 2008 to December 2017were extracted from the China Information System for Disease Control and Prevention. An active surveillance of C. sinensis infection in fish was conducted in 2016-2017. County level data including potential environmental, social-economical and behavioral determinants was also collected. Univariate and multivariate logistic regression models were used to explore the determinants of C. sinensis infection in humans and fish. Simple and multiple zero-inflated Poisson regression models were fit to assess the associated factors of clonorchiasis in humans at the county level., Results: Totally, 4526 C. sinensis cases were reported between 2008 and 2017, with an annual prevalencerate of 0.96/100,000 persons. Of 101 counties in Guangxi, 97 reported at least 1 case. Among 2,098 fish samples, 203 (9.7%) from 70 counties contained C. sinensis. The rate was higher in small fish including Pseudorasbora parva (45.3%), Misgurnus anguillicaudatus (41.2%), Hemicculter leuciclus (34.5%), unclassified small fishes (30.9%), Cyprinidae (20.0%), Cirrhinus molitorella (16.4%), Carassius auratus (13.6%) and Cyprinus carpio (13.3%), while it was lower in fish species that are usually used in preparing raw fish dishes including Ctenopharyngodon idellus (3.6%), Spinibarbus denticulatus (3.7%), Monopterus albus (6.4%), Cyprinus carpio (4.4%), Oreochromis mossambicus (3.3%) and Spualiobarbus Curriculus (6.6%). The C. sinensis infection in fish was only associated with fish species. The estimated human clonorchiasis prevalence at the county level was positively associated with raw fish consumption habits and certain rivers., Conclusions: Clonorchiasis is highly prevalent in both humans and freshwater fish in Guangxi. Environmental, social-economic and behavioral determinants contribute to the high prevalence as well as the significant differential distribution by county. Regular surveillance should be implemented for clonorchiasis to demonstrate the change in epidemiology and burden, which will benefit the design of interventions., (© 2022. The Author(s).)

Background: To understand Clonorchis sinensis reinfection and the determinants of reinfection in endemic areas is important in establishment of control measures., Methods: A prospective cohort study was implemented in Hengxian County, Guangxi, China. Individuals with C. sinensis infection were completely treated, and those cured were enrolled as study subjects and followed up for 3, 6, and 12 months. The reinfection frequency and incidence were calculated, and a multivariable Cox proportional hazard model was constructed to capture reinfection determinants., Results: Among 635 enrolled subjects, 436 (68.7%) completed follow-up. Of these, 177 (40.6%) were reinfected; 133 (75.1%) were reinfected once, 41 (23.2%) twice, and 3 (1.7%) three times. The incidence of reinfection was 64.0 per 100 person-years. Men (adjusted hazard ratio [aHR], 1.67; 95% confidence interval [CI], 1.14-2.44), those with underlying diseases (aHR, 1.41; 95% CI, 1.02-1.95), and those with moderate- or heavy-intensity infections (aHR, 1.45; 95% CI, 1.14-1.85) had increasing reinfection probabilities., Conclusions: C. sinensis reinfection is high in endemic areas. Men and high-intensity infection are important determinants of reinfection. Repeated chemotherapy is necessary to control reinfection and its associated morbidities, especially in high-risk individuals. In addition, behavioral education is advised to decrease overall reinfection in endemic areas., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Clonorchiasis is recognized as an important zoonotic parasitic disease worldwide. However, the roles of host long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) in the response to Clonorchis sinensis ( C. sinensis ) infection remain unknown. Here we compared the expression of lncRNAs and mRNAs in the liver tissue of mice infected with C. sinensis , in order to further understand the molecular mechanisms of clonorchiasis. A total of 388 lncRNAs and 1,172 mRNAs were found to be differentially expressed with absolute value of fold change (FC) ≥ 2.0 and p < 0.05 by microarray. Compared with controls, Gm6135 and 4930581F22Rik were the most over- and under-expressed lncRNAs; flavin-containing monooxygenase 3 ( Fmo3 ) and deleted in malignant brain tumors 1 ( Dmbt1 ) were the most over- and under-expressed mRNAs. Moreover, functional annotation showed that the significantly different mRNAs were related with "FOXO signaling pathway", "Wnt signaling pathway", and "AMPK signaling pathway". Remarkably, lncRNA Gm8801 were significantly correlated with mRNA glycerol-3-phosphate acyltransferase mitochondrial ( Gpam ), insulin receptor substrate 2 ( Irs2 ), and tumor necrosis factor receptor superfamily member 19 ( Tnfrsf19 ) in ceRNA networks. These results showed that the expression profiles of lncRNAs and mRNAs in the liver changed after C. sinensis infection. Our results provided valuable insights into the lncRNAs and mRNAs involved in clonorchiasis pathogenesis, which may be useful for future control strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Han, Zhang, Jiang, Li, Ding, Zuo, Duan, Chen, Sun, Hu, Gao and Zhang.)

The Greater Mekong Subregion (GMS) is a trans-national region of the Mekong River basin in Southeast Asia comprising Cambodia, the People's Republic of China (specifically Yunnan Province and Guangxi Zhuang Autonomous Region), Lao People's Democratic Republic (Lao PDR), Myanmar, Thailand, and Vietnam home to more than 340 million people or almost 4.5% of the global population. Human liver fluke infections caused by Opisthorchis viverrini and Clonorchis sinensis have been major public health problems in this region for decades. Opisthorchiasis caused by O. viverrini is prevalent in Thailand, Lao PDR, Cambodia and central-southern Vietnam with more than 12 million people infected. Clonorchiasis caused by C. sinensis is endemic in northern Vietnam and Guangxi with estimated 3.5 -5 million infected. The infections can cause several liver and biliary diseases such as cholangitis, periductal fibrosis, gallstones, and cholangiocarcinoma (CCA), a fatal bile duct cancer. Key determinants of the geographical distribution differences of the two liver fluke species are snail species and geographic barriers. Main risk behaviour of the infections is the culture of eating raw fish "the raw attitude" of people in the GMS, especially the Tai/Dai/Thai/Laos ethnic groups, the major population in the GMS. Over the past 20 years, there is a big change in prevalence of the infections. Opisthorchiasis has long been endemic, particularly in northern and northeastern Thailand and Lao PDR with over 8-10 million cases estimated. However, after several rounds of national campaign against opisthorchiasis using integrated control approach in Thailand over the past three decades, the prevalence of O. viverrini infection has reduced from over 15% in 1996 to 2.2% in 2019. High prevalence of O. viverrini infection continues in Lao PDR and central Vietnam. Emerging high prevalence, up to a maximum of 47.5%, has been noted in Cambodia during the past 10 years possibly due to more studies being conducted rather than increasing prevalence. O. viverrini infection has now also been reported in Lower Myanmar in recent years. Clonorchiasis has been known in northern Vietnam and southern China for a long time. Several surveys have reported infections in Guangxi in the last 10 years, and until now liver fluke infected cases have not been reported in Yunnan. Overall, nowadays, there is a shift in high risk areas for GMS liver fluke infection from northeastern Thailand to Lao PDR, Cambodia, Vietnam, Myanmar, and Guangxi P.R. China. Urgent systematic disease mapping and integrated liver fluke control using One Health approaches should be implemented nationwide in GMS countries., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Extensive parasitic diseases epidemiology in Zhejiang province has not been carried out since the second national survey in 2004. Therefore, dynamics in prevalence and infection pattern of the major intestinal parasites should be explored., Methods: The distribution of three parasites including soil-transmitted helminths (STH), intestinal protozoa and C. sinensis in Zhejiang from 2014 to 2015 were explored. Kato-Katz technique was used for STH and C. sinensis detection, whereas transparent adhesive paper anal swab was used for pinworm detection, and iodine smear was used for protozoa detection. A questionnaire survey on alimentary habits and sanitary behaviors was conducted in half of the studied counties., Results: This study recruited 23,552 participants: 19,935 from rural and 3617 from urban area. Overall prevalence of intestinal helminth infections was 1.80%. In this study, seven helminth species were identified including A. duodenale, N. americanus, Trichuris trichiura, Ascaris lumbricoides, C. sinensis, Fasciolopsis buski and pinworm. The average prevalence of STH infection was 1.71%: 1.94% in rural and 0.44% in urban area. Hookworm was the most prevalent infection at 1.58%: 1.79% in rural and 0.44% in urban area. Prevalence varied considerably in the studied counties. Prevalence was highest in Yongkang county at 10.25%. Only 2.79% of children from rural area were infected with pinworm. A proportion of 0.40% of rural participants were infected with protozoa, whereas Endolimax nana was the most prevalent at 0.23%. C. sinensis showed infection only in one man. Awareness on C. sinensis was 24.47% in rural and 45.96% in urban area, respectively., Conclusions: Prevalence of STH and protozoa infections declined considerably whereas C. sinensis infections remained few in Zhejiang province compared with the prevalence reported in previous large scale surveys (19.56% for national STH infection in 2004, 18.66% and 4.57% for provincial STH and protozoa infection, respectively in 1999). The findings of this study showed that hookworm, mainly N. americanus remained a parasitic threat to population health, mainly in the central and western Zhejiang. Therefore, more health education regarding fertilization and farming habits is necessary in rural areas. The awareness concerning hookworm infection should be reinforced., (© 2021. The Author(s).)

Background: Clonorchis sinensis (CS) infection is considered a group 1 carcinogen of cholangiocarcinoma (CCA). There were very few studies regarding clinical characteristics of CS-associated CCA (CACC). This study aimed to investigate clinical characteristics of patients with CCA with or without CS infection. Methods: A total of 367 patients diagnosed with CCA who underwent diagnostic tests for CS infection were enrolled. CS infection was defined as follows: at least one positive serum ELISA test, skin test, stool microscopy, or bile microscopy. Results: There were 95 (26%) patients with CS infections. The median follow-up duration was 14.9 months (range, 6.07-36.17). The following significant differences were noted among patients with CACC compared to non-CACC; diagnosis at younger age (median 62 years vs. 65 years, p = 0.018), higher male to female ratio (83.2 vs. 61.8%, p < 0.001), and residence in CS-endemic area (46.3 vs. 25.4%, p = 0.014). Univariate analysis of prognostic factors indicated that tumor location, curative resection, tumor stage, and laboratory tests including CA 19-9, CEA, and bilirubin were significantly associated with overall survival, but CS infection was not. In multivariate analysis, tumor location, CEA, curative resection and tumor stage were identified as independent prognostic factors. Among patients under age 64, CACC group had lower survival rate than non-CACC group ( p = 0.022). Conclusions: CACC had the following significant characteristics compared to non-CACC; diagnosis at younger age, higher male to female ratio, higher prevalence in CS endemic areas and poorer overall survival in patients under age 64., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Chang, Lee, Kim, Yang, Park, Choi, Kang, Lee, Lee, Lee, Park and Park.)

Clonorchiasis is an important food-borne parasitic disease in China because of the popularity in ingesting raw freshwater fish. To explore the epidemiology and determinants of clonorchiasis in children, a cross-sectional survey was implemented in two middle schools in Qiyang county, Hunan province, in southeastern China. Questionnaire survey and fecal examination were implemented. Questionnaires were fed back by 627 students, while stool samples were collected from 557 students, out of which 545 ones also provided questionnaire information. The percentage of students ingesting raw freshwater fish was 40.5% (254/627), while the prevalence with Clonorchis sinensis infection was 18.9% (105/557). Such factors contributed significantly to the students' practice in eating raw freshwater fish including boys, fathers' eating raw freshwater fish, mothers' eating raw freshwater fish, and preparation of raw freshwater fish at home, with an adjusted odds ratio of 1.9 (95% confidence intervals (95% CI): 1.3-2.8), 3.9 (95% CI: 2.3-6.5), 3.0 (95% CI: 1.8-4.8) and 2.8 (95% CI: 1.8-4.5), correspondingly. Ingestion of raw freshwater fish, fathers' eating raw freshwater fish and preparation of raw freshwater fish at home were risk factors of C. sinensis infection in students, and the adjusted odds ratio was 3.2 (95% CI: 1.9-5.5), 2.1 (95% CI: 1.1-3.9) and 1.8 (95% CI: 1.0-3.2), respectively. Thus, clonorchiasis was endemic in the surveyed schools due to the ingestion of raw freshwater fish, which is influenced by family environment. Education should be implemented in schools to promote behavioral change of eating raw freshwater fish., (Copyright © 2020. Published by Elsevier B.V.)

Several foodborne zoonotic trematodes of the family Opisthorchiidae have been reported to infect people, specifically two genera, Clonorchis and Opisthorchis. Three species Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini are the most extensivley studied of the Opisthorchiidae. At least 680 million people worldwide are at risk of infection of these liver flukes through the consumption of raw or partially cooked freshwater cyprinid fish. An estimated 45 million people in Europe and Asia are currently infected with these liver flukes. Of these, the 35 million are infected with C. sinensis mainly in China, 10 million with O. viverrini in Southeast Asia, and 1.2 million with O. felineus in Eastern Europe and Russia. These liver flukes have been proven to be causative agents of bile duct cancer or cholangiocarcinoma (CCA). A multidisciplinary program should be implemented involving comprehensive research on molecular genetics, diagnosis, treatment, prevention, as well as educational and control programs should progressively be introduced and applied in endemic regions of O. viverrini, O. felineus and C. sinensis throughout their ranges, hence, opisthorchiasis and clonorchiasis free communities can be realised globally., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Stool examination is the gold standard for the detection of intestinal parasites. We assessed the performance of a newly developed AVE-562 analyzer (AVE Science & Technology Co., Hunan, China) for the vision-based detection of eggs of Clonorchis sinensis -the most common intestinal parasite in Korea-in stool samples. In total, 30 stool samples with a high or low egg count or without eggs (as negative control samples) (N=10 each) were prepared and analyzed. The performance of the AVE-562 analyzer was compared with that of the formalin-ether concentration (FEC) method. The overall correct identification rate of the AVE-562 analyzer based on FEC results was 66.6%. The sensitivity, specificity, positive predictive value, and negative predictive value of the AVE-562 analyzer for detecting C. sinensis eggs were 36.4%, 100.0%, 100.0%, and 73.1%, respectively. The average time required to run five tests simultaneously was 27 min using the AVE-562 analyzer and 58 min using the FEC method. Although the AVE-562 analyzer enables rapid and convenient stool examination, its sensitivity needs to be improved, particularly considering the prevalence of low-burden C. sinensis infection in Korea.

Soil-transmitted helminthiases (STHs) are caused by a group of intestinal nematode infections due to poor hygiene and environments, and clonorchiasis is a food-borne trematode (FBT) infection caused by ingestion of raw freshwater fish. Both are endemic in the People's Republic of China. To explore a suitable control strategy, integrated interventions were applied between 2007 and 2009 in ten pilot counties (eight for the STHs and two for clonorchiasis). Drug administration was used for treatment and complementary efforts to improve the situation based on health education, provision of clean water and sanitation were carried out. Significant achievements were gained as reflected by a drastic decrease in prevalence these infections were demonstrated. The overall prevalence of STHs and clonorchiasis decreased from 35.9% to 7.8% and from 41.4% to 7.0%, respectively. The reduction of prevalence and high cost-effectiveness were documented supporting large-scale application of this integrated intervention in China and elsewhere., (Copyright © 2020. Published by Elsevier B.V.)

To effectively promote the implementation of interventions, the identification of high-risk groups and the characteristics of Clonorchis sinensis infection in endemic regions are needed. In a clonorchiasis-endemic area, local residents were randomly enrolled for helminth egg examination in June 2016. The prevalence in subpopulations as well as the knowledge, attitudes, and behaviours and the factors influencing clonorchiasis in egg-positive populations were analysed. A total of 2282 local residents participated in the survey; the C. sinensis prevalence was 48.6% (1109 persons). A higher prevalence was found in males (62.6%) than in females (29.7%). People older than 30 years had the highest prevalence (52.7%-57.6%). Among the 888 persons who were infected with C. sinensis and participated the questionnaire investigation, 19.0% (169/888) knew that it could cause cancer. In addition, 60.6% of people reported that they intended to keep eating raw fish despite knowing the risk of infection. The two primary reasons for continuing to eat raw fish were the disease being regarded as not serious (38.3%) and the belief that anti-parasite medications are effective (39.6%). A total of 94.4% (797/844) of responders reported eating raw fish more frequently in the home than outside of the home. Our study revealed a notably high C. sinensis prevalence in the study area. Awareness of clonorchiasis disease severity should be increased among high-risk individuals and families in highly endemic areas., Competing Interests: The authors declare that they have no conflict of interest., (© 2020 Published by Elsevier Inc. on behalf of International Association of Food and Waterborne Parasitology.)

Mice with different genetic backgrounds have various susceptibilities to infection with Clonorchis sinensis , although the mechanisms underlying are largely unknown. Toll-like receptor 4 (TLR4) as one of the most important pattern recognition receptors (PPRs) is essential for the invasion, survival, pathogenesis, and elimination of worms. The roles played by TLR4 in C. sinensis infection may vary due to the different genetic backgrounds of mice. In the present study, a relatively resistant mouse strain-C57BL/10 to C. sinensis was used for investigation on the possible roles of TLR4 in the biliary injuries and peribiliary fibrosis. TLR4 wild type (TLR4 wild ) and TLR4 defective (TLR4 def ) mice were orally infected with 45 metacercariae of C. sinensis , and all C. sinensis -infected mice and non-infected groups were anesthetized on day 28 post-infection. The liver and serum from each mouse were collected for assessment of the biliary injuries and biliary fibrosis. Meanwhile, hepatic leukocytes were isolated and detected for the activation of M1 or M2 macrophage using flow cytometry. The hepatic type 1 immune response and type 2 immune responses -relative molecules were also evaluated using ELISA and quantitative PCR. The data showed that TLR4 def aggravated liver inflammatory cell infiltrations, bile duct proliferation, biliary and hepatocellular injuries, and ECM deposition in C. sinensis -infected mice, compared with TLR4 wild mice when they were intragastrically administered with the same amounts of C. sinensis metacercaria. Furthermore, the M2-like macrophages and type 2 immune responses were significantly predominant induced in TLR4 def mice, compared with that of TLR4 wild mice following C. sinensis infection. But the type 1 immune response were significantly decreased in TLR4 def mice, compared with TLR4 wild mice after C. sinensis infection. These data demonstrate that TLR4 deficiency exacerbates biliary injuries and peribiliary fibrosis caused by C. sinensis in C57BL/10 strain mice, which is contributed by augments of type 2 immune responses and decrease pro-inflammatory responses., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Yan, Wu, Xu, Li, Zhou, Yang, Cheng, Liu, Dong, Koda, Zhang, Yu, Chen, Tang and Zheng.)

Objectives: Immune responses are tightly regulated in the development of clonorchiasis. However, the adaptive immune regulatory pathway contributing to the pathological processes of clonorchis sinensis (C. sinensis) infection is still not clear. In this study, we assessed the dynamic changes of CD4 + T cell subsets and the related cytokines as well as transcription factors during C. sinensis infection., Materials and Methods: We used female FVB mice to establish the infection model. H&E and Masson's stain were performed in 2 or 8 weeks post-infection (PI) liver of C. sinensis . The percentages of splenic Th1, Th2, and Treg in CD4+T cells were detected by flow cytometry. The transcription factors T-bet, GATA3, Foxp3, and RORγt gene expression were detected by qPCR. The protein expression of IL-10, IL-17, IL-4, IL-2, and Tumor Necrosis Factor-α (TNF-α) were examined using ELISA., Results: The percentages of splenic Th1, Th2, and Treg in CD4 + T cells were significantly increased in both 2 and 8 weeks PI of C. sinensis , while the ratio of Treg/Th17 as well as the percentage of Treg in serum was gradually increased during the development of infection. The expressions of T-bet, GATA3, Foxp3, and RORγt were increased in 8 weeks PI. Serum levels of IL-10, IL-17, IL-4, and IL-2 were profoundly increased in infected mice, while the concentrations of TNF-α increased to peak two weeks PI., Conclusion: Our results suggested that the imbalance of CD4 + T cell subsets may regulate and contribute to an appropriate compromise between pathology, tissue repair, and elimination in a susceptible murine host.

Liver flukes ( Fasciola spp., Opisthorchis spp., Clonorchis sinensis ) and blood flukes ( Schistosoma spp.) are parasitic helminths causing neglected tropical diseases that result in substantial morbidity afflicting millions globally. Affecting the world's poorest people, fasciolosis, opisthorchiasis, clonorchiasis and schistosomiasis cause severe disability; hinder growth, productivity and cognitive development; and can end in death. Children are often disproportionately affected. F. hepatica and F. gigantica are also the most important trematode flukes parasitising ruminants and cause substantial economic losses annually. Mass drug administration (MDA) programs for the control of these liver and blood fluke infections are in place in a number of countries but treatment coverage is often low, re-infection rates are high and drug compliance and effectiveness can vary. Furthermore, the spectre of drug resistance is ever-present, so MDA is not effective or sustainable long term. Vaccination would provide an invaluable tool to achieve lasting control leading to elimination. This review summarises the status currently of vaccine development, identifies some of the major scientific targets for progression and briefly discusses future innovations that may provide effective protective immunity against these helminth parasites and the diseases they cause.

Clonorchis sinensis could induce inflammation, epithelial hyperplasia and fibrosis in the intrahepatic bile duct as a food-borne parasite, which was associated with the development of cholangiocarcinoma (CCA). Praziquantel was the most effective drug on treatment of this kind of parasite. However, new drugs with minimal toxicity to the host were urgently needed due to the side effects of Praziquantel and its CCA risk. In this study, helminth mitochondria respiratory chain blocker Wortmannilatone F (WF) and IL-8 analogue CXCL8 (3-72) K11R/G31P were used to treat BALB/C mice infected by Clonorchis sinensis. We investigated the gross and histopathological morphology of the liver, inflammation-associated cytokine IL-6, lipid peroxidation-related proteins cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX), collagen fiber accumulation and fibroblast-specific protein 1 (FSP1), malignant markers proliferating cell nuclear antigen (PCNA) and cytokeratin 19 (CK19), as well as the disinfection effect on these parasites in vitro. WF inhibited and killed the worms dramatically, and the combination of WF with G31P improved the condition of the hepatobiliary duct tissue greatly. These outcomes indicated that the combination of WF and G31P was a potential therapeutic method to treat the Clonorchis sinensis infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Clonorchiasis is caused by the ingestion of raw freshwater fish containing Clonorchis sinensis larvae. Health education is crucial in sustainable control of clonorchiasis but extremely challenging when dealing with adults. To investigate if education in children would be a beneficial long-term approach, a cluster-controlled trial was implemented in two primary schools between 2017 and 2019 in Binyang county, Guangxi, China. A baseline survey was conducted in both schools, and then comprehensive education activities were implemented in the interventional one. Six and 12 months later, all pupils were subjected to a survey to assess the potential change of knowledge, practice and belief related to clonorchiasis. Finally, 247 pupils in interventional school and 151 ones in control were eligible for analysis. In the interventional school, the percentage of pupils with knowledge on transmission route, early symptoms, complications and carcinogenicity increased from 15.0% to 78.5% (p < 0.001), from 6.5% to 36.8% (p < 0.001), from 4.0% to 93.9% (p < 0.001) and from 4.5% to 49.4% (p < 0.001) after 12 months. In this school, 16 pupils (6.5%) had eaten raw freshwater fish during the year immediately before the baseline survey, a number that decreased to 3 (1.2%) (p = 0.002) 12 months later. The percentage of students had the belief on the ability not to eat raw freshwater fish in the future was 70.9% at the baseline and 97.2% 12 months later (p < 0.001). Although knowledge related to clonorchiasis also increased in the control school, the absolute percentage of changes was lower; in particular, practice and belief had not changed., Competing Interests: Declaration of Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to investigate the epidemic scope, epidemic pattern and degree of damage of clonorchiasis as well as to summarize the diagnosis and control effect of clonorchiasis in Shandong Province, China., Methods: A retrospective study was conducted to summarize the research on the dynamic epidemiology and the prevention and treatment measures of clonorchiasis in Shandong Province, China from 1962 to 2015., Results: After unremitting efforts and three nationwide surveys on the distribution and infection of human parasites, the infection rate of clonorchiasis decreased from 1.51% (1,500, 000) in the1960s to 0.16% in 1991, 0.04% in 2000 and 0.007% in 2015, respectively. The infection rate for villages with perennial water accumulation and rainy season water accumulation was higher than that of villages with rainy season water accumulation and non-water accumulation. The egg flotation in 70% potassium iodide solution method was applied to accurately test the specific proportion of C. sinensis eggs, which was 1.42. Using the modified Kato thick smear method, the detection rate of eggs was over 98%; hexachloroparaxylene yielded satisfactory therapeutic effects with slight adverse reactions, it is effective in the treatment of clonorchiasis., Conclusions: After 50 years of active prevention and treatment, the disease has been successfully controlled in Shandong Province and had been used for reference in other endemic areas of clonorchiasis in China., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Hepatosplenic candidiasis and other fungal infections of the liver are uncommon in healthy individuals; however, high index of suspicion is essential in immunocompromised patients with prolonged fever. Parasitic infections are protozoan or helminthic; their distribution and epidemiology are variable among different world regions. Clonorchiasis, opisthorchiasis, fascioliasis, and ascariasis are helminthic infections that commonly involve the biliary systems. Signs and symptoms of cholangitis require prompt management to relieve biliary obstruction; addition of antihelminthic agents is essential. Parasitic infections are mostly transmitted to humans by fecally contaminated food and water. Proper hand and food sanitation measures are essential in preventing disease transmission., Competing Interests: Disclosure Nothing to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Clonorchiasis is endemic in East and Southeast Asian countries. For a preventive strategy against infectious diseases, vaccination is the most effective. Here, we evaluated the molecular characteristics and immune responses of CsAg17 protein from Clonorchis sinensis, and investigated its protective effects against C. sinensis challenge., Methods: A cDNA clone encoding CsAg17 protein and containing a secretory signal peptide at the N-terminus was retrieved from the C. sinensis transcriptome bank. Recombinant CsAg17 B-cell epitope protein and cDNA vaccines were produced and their immune responses were evaluated in FVB mice. The proportional changes of CD3 + /CD4 + and CD3 + /CD8 + T cells were detected by flow cytometry, and immune effectors were measured by ELISA., Results: The CsAg17 mRNA was transcribed at a higher level in C. sinensis adults than in metacercariae. The CsAg17 protein was distributed in the sperms, oral and ventral suckers, and mesenchymal tissues of C. sinensis adults. In mice challenged with C. sinensis metacercariae, vaccination with CsAg17 protein and cDNA resulted in a reduction to 64% and 69% in worm burden, respectively. Both CsAg17 protein and cDNA vaccines increased the proportion of CD3 + /CD4 + and CD3 + /CD8 + T cells and stimulated the production of Th1 type cytokines such as interleukin (IL)-2, IL-12, and interferon-γ, while maintaining minimum levels of Th2 cytokines. The levels of IgG specific to CsAg17 protein steeply increased in the two vaccinated groups from 2 weeks after immunization. The liver tissue retained good morphology in the mice vaccinated with CsAg17 protein or cDNA, whereas severe inflammation and large serous cysts were observed in the liver of the unvaccinated mice., Conclusions: Vaccination with CsAg17 protein and cDNA reduced the pathological changes in the bile duct and liver, and ameliorated the worm burden via cellular and humoral immune responses. Thus, they may serve as good vaccine candidates against C. sinensis infections.

Clonorchiasis is an important food-borne parasitic disease in China, and infection with C. sinensis can cause hepatobiliary diseases. Comprehensive and systematic prevention and control of clonorchiasis requires the establishment of an effective surveillance. A total of 301 surveillance points were set up in 30 provinces across China in 2016, and 1000 people were selected by cluster sampling at each surveillance point annually for C. sinensis infection screening using Kato-Katz thick smear method. C. sinensis infection was detected in 6226 people following screening of 305081 people at the 301 surveillance points in 2016. Infection rate was 2.04%; with C. sinensis infection detected in 70 counties spread across 15 provinces, 89.37% of the infected people were distributed in Jilin, Heilongjiang, Guangdong and Guangxi provinces. Highest infection rate was observed in Da'an city, Jilin Province (49%). The national infection rate in male and female was 2.70% and 1.40% respectively. Infection rate between male and female was significantly different (P <0.01). Disease prevalence increases with age in both male and female, reaches peak in age group 40-49. Result obtained indicate that major C. sinensis endemic areas are distributed in the north and south of China, and areas with high prevalence are distributed along the river system at county level. Result, also, shows that middle-aged men are at high-risk of infection. These results suggest that surveillance activities should be sustained nationwide and highlight the need for an integrated approach to control C. sinensis transmission in regions with high disease prevalence in China., Competing Interests: Declaration of Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Clonorchiasis is caused by ingestion of raw freshwater fish containing infective larvae of Clonorchis sinensis. China harbours the largest number of people with C. sinensis infection. During the past three decades, the National Institute of Parasitic Diseases, affiliated to the Chinese Center for Disease Control and Prevention (NIPD) conducted many studies to facilitate the control on clonorchiasis. Three national surveys have shown the updated epidemiology of clonorchiasis in China. Recently, a national surveillance system has also been established, which will enable the production of high-resolution map. The evaluation of the disease burden has enhanced the awareness on clonorchiasis. Diverse diagnosis techniques including rapid screening by questionnaire, serological tests, faecal examination and a molecular method have been developed or evaluated. The NIPD also participated in the early evaluation of praziquantel against clonorchiasis, which enhanced its application in China. Also, the NIPD has verified the efficacy of tribendimidine against clonorchiasis. A new sustainable strategy is also being explored. However, more research is expected to further facilitate control of clonorchiasis in China, as well as international cooperation in fighting human liver fluke infections in Asia., (© 2020 Elsevier Ltd. All rights reserved.)

Intraductal papillary neoplasm of the biliary tract (B-IPN) is a scarcely known entity in our daily practice due to its low prevalence. Until its new definition in the fourth edition of the WHO classification of the digestive tract tumors of 2010 the disease was grouped under a heterogeneous and imprecise terminology. In addition, in recent years there has been progress in the knowledge of its etiopathogenesis, its natural history and its findings in image. The purpose of this paper is to review these data underlining the radiological findings of the disease and its differential diagnosis., (Copyright © 2019 SERAM. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: Clonorchiasis, caused by Clonorchis sinensis (C. sinensis) infection, is a serious food-borne zoonotic disease that is often asymptomatic or shows only mild symptoms, which leads to delayed treatment and chronic clonorchiasis and results in various complications, such as cholelithiasis, cholangitis, cholecystitis and cholangiocarcinoma. However, acute shock caused by C. sinensis infection has not been reported. Here, for the first time, we describe a fatal case of acute shock caused by C. sinensis infection., Case Presentation: A patient with a history of eating raw or undercooked freshwater fish was hospitalized with acute shock caused by severe abdominal pain. Physical examination suggested acute abdomen with severe abdominal pain and rigidity. Computed tomography (CT) detection indicated acute cholecystitis and cholelithiasis. After cholecystectomy, several liver flukes were found in the drainage tube. Furthermore, morphological analysis and polymerase chain reaction (PCR) identified the pathogen as C. sinensis. The liver gradually restored normal function after anthelmintic therapy with praziquantel., Conclusions: In this fatal case, C. sinensis infection was the cause of acute shock, which is rarely found in the clinic environment. This report aims to increase awareness of the hazards and complications related to clonorchiasis. The PCR diagnosis method used in this report might be helpful in reducing the misdiagnosis of clonorchiasis and unnecessary cholecystectomy.

Background: Clonorchiasis is caused by eating of raw or undercooked freshwater fish containing the larvae of Clonorchis sinensis; the Kato-Katz method is widely applied in diagnosis. The improvement of repeated Kato-Katz smears from multiple stool samples has been well illuminated in many helminths other than C. sinensis., Methods: A cross-sectional investigation was implemented to capture the epidemiology and risk factors of clonorchiasis among middle school students in Qiyang county, China. Students with complete data of six Kato-Katz thick smears from two stool samples were included in this analysis. Data on the habits of eating raw freshwater fish were also collected and compared., Results: Altogether, 397 students had complete information of six smears, out of which 394 reported the information on eating habits. According to the 'gold' standard by six smears, 77 students (19.4%) were detected with C. sinensis. However, only 45 (11.3%) were detected using a single smear, with an underestimation of 41.6% compared to the 'gold' standard. However, the geometric mean of eggs per gram of feces in detected cases was 126.4 in a single smear, overestimated by 105.2% compared to 61.6 by the 'gold' standard. The linear relationship between prevalence and infection intensity of detected cases based on different smears was significantly negative. The habits of eating raw freshwater fish in the false negative cases was similar to those in the detected cases, but these two groups had significantly higher levels for habits of eating raw freshwater fish than negative individuals., Conclusions: In low endemicity situations, underestimation of C. sinensis infection could not be avoided based on a limited number of Kato-Katz smears. Thus, repeated smears from at least two stool samples should be considered when an individual eats raw freshwater fish, drug efficacy is evaluated or elimination of C. sinensis is verified. Additionally, when logistics are insufficient for multiple samples to be taken for diagnosis for survey and surveillance in the areas or populations of low endemicity, prevalence accuracy needs to be corrected.

Objective: To assess the clinical significance of transient elastography (Fibroscan) in detection of clonorchiasis, so as to provide new insights into the assessment of therapeutic efficacy of deworming., Methods: The liver stiffness measurement (LSM) values were measured in parasitologically diagnosed clonorchiasis patients using FibroScan before and after deworming, and the patients'age, gender, body mass index (BMI), duration of raw fish consumption and total amount of raw fish consumption were collected for correlation analyses., Results: The clonorchiasis patients'age, gender, BMI, duration of raw fish consumption and total amount of raw fish consumption had no associations with pre-treatment LSM values ( r / r s = 0.189, 0.073, 0.180; 0.071, -0.098, 0.033; 0.166, 0.309, 0.172; 0.235, 0.247, 0.209; 0.164, 0.277, 0.088; all P values > 0.05). There was a significant difference in the LSM values from the seventh, eighth and ninth intercostal space prior to deworming ( F = 3.259, P < 0.05), and no significant difference was detected after deworming ( F = 0.851, P > 0.05). The LSM values from the seventh, eighth and ninth intercostal space were significantly lower pre-deworming than post-deworming ( t = 6.724, 5.603, 2.884; all P values < 0.05)., Conclusions: FibroScan is feasible to assess the therapuetic efficacy of deworming in patients with clonorchiasis; however, measurement at various sites affects the LSM value.

A clonorchiasis case in a captive leopard cat, Prionailurus bengalensis euptilurus, was confirmed by ultrasonographic findings and egg morphologies found in the bile juice sample in the Korea. The leopard cat was introduced from the wild habitat of Gyeongsangnam-do, to Cheongju Zoo in Cheongju-si, Chungcheongbuk-do, Korea in August 2014. Physical examinations were basically performed for quarantine and check-up health. The cat was comparatively good in health except anorexia. The cyst-like bile duct dilation and the increased echogenicity of gall bladder wall and hepatic parenchyma were observed by ultrasonography. Ultrasound-guided needle biopsy was conducted for collecting bile juice and the specimens were observed under light microscope. The numerous small trematode eggs were detected in the bile juice sample of the light microscopy. The eggs were 25-33 (28±3) μm by 18-22 (20±1) μm in size and showed typical characteristics of Clonorchis sinensis egg, i.e., a dominantly developed operculum, shoulder rim and dust-like wrinkles in surface. To treat the liver fluke infection, 20 mg/kg of praziquantel was orally administered only once to the case. Follow-up studies including fecal examinations were conducted during 2 years after treatment. But no more eggs were detected from the case. In the present study, we described the first clonorchiasis case of leopard cat, which was confirmed by ultrasonographic findings and egg morphologies from the bile juice sample in Korea.

Background: Foodborne Neglected Zoonotic Helminths (FNZH) are parasites of both economic and public health importance. They include Taenia solium, Echinococcus granulosus sensu lato, Echinococcus multilocularis and Foodborne trematodes (FBT). FNZH are earmarked for major interventions for control, elimination and eradication. This systematic review highlights the progress towards development of rapid tests for the diagnosis of FNZH since 2010 when they were listed as neglected tropical diseases., Methodology: A systematic search was conducted in three databases, World of Science, Embase and PubMed using the same search phrase. The search produced 480 hits. Three studies from back referencing were included. Only 22 of these met the inclusion criteria. Data was extracted from these and presented qualitatively., Results: Twenty-five rapid diagnostic tests were found to have been developed since 2010, eight for diagnosis of T. solium infections, eight for echinococcosis and nine for FBT infections. The rapid tests for diagnosing T. solium infections included six antibody detecting and two antigen detecting tests. They constitute a combination among them, with some tests providing qualitative, others quantitative results. Similarly, seven out of the eight rapid tests developed for Echinococcus infections were antibody detecting tests save for one loop mediated isothermal amplification test. All of them were qualitative tests. For FBT infections, nine rapid tests were described; two antibody and one nucleic acid detecting test for diagnosis of Fascioliasis; three nucleic acid detecting tests for Opisthorchiasis; one antibody detecting test for Paragonimiasis; and for Clonorchiasis, one antibody and one nucleic acid detecting test. The FBT infection rapid tests were all qualitative in nature. Most of these tests have not undergone field evaluation in endemic areas where they will be used most., Conclusion: This review describes the development and evaluation of rapid diagnostic tests, while highlighting the need for in depth validations of the tools to determine how well they can perform in endemic areas., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To understand the status and capability of professionals at provincial parasitic diseases control institutions in main clonorchiasis-endemic areas of China., Methods: The status and capacity of professionals at four provincial parasitic diseases control institutions were collected using questionnaire surveys, including Guangdong, Guangxi, Heilongjiang and Jilin, and the data were analyzed., Results: There were totally 37 professionals working on parasitic diseases control in the four provincial institutions in 2018, including 33 full-time and 4 part-time professionals, and there were 12, 16, 3 and 6 professionals working at Guangdong, Guangxi, Heilongjiang and Jilin provincial institutions, respectively. Of the 37 professionals, there were 24.32%, 37.84% and 37.84% at ages of 35 years and lower, 35 to 45 years, and greater than 45 years. Men consisted of 54.05% of all professionals, and 86.49% had an education of bachelor and above, while 78.38% had a title of moderate or above. Among all professionals, there were 29 responsible for clonorchiasis prevention and control; however, they all participated in the prevention and control of other parasitic diseases; 33 professionals were able to prepare Kato-Katz smears and 34 read the smears; 30 professionals were able to detect metacercaria in fishes, and 24 and 16 professionals were able to perform immunological and mo-lecular tests. In addition, 26 professionals participated in provincial projects, 19 in national projects; however, few professionals participated in international projects or undertook provincial, national or international projects. Furthermore, there were 34 professionals (91.89%) participating in national trainings on parasitic diseases; however, only 12 (32.43%) completed a training for more than one week., Conclusions: There are few professionals at provincial parasitic diseases control institutions in main clonorchiasis-endemic areas of China, and they have a relative high capability in parasitic disease control; however, their research capacity is relative weak.

Purpose: Parasite infestation (PI) is reportedly related to intrahepatic stones (IHSs) and common bile duct (CBD) stones. This study assessed the association of clonorchiasis and helminthiasis with the sub-types of cholelithiasis., Methods: The study cohort consisted of 713 patients who were aged ≥20 years with clonorchiasis or helminthiasis for the first time between 2000 and 2010 from the National Health Insurance Research Database. The controls without clonorchiasis and helminthiasis were randomly selected with a 1:1 propensity score matching. All patients were followed up until the end of 2011 to measure the incidence of cholelithiasis., Results: The risks of cholelithiasis in the clonorchiasis (adjusted hazard ratio [aHR] = 3.72, 95% confidence interval [CI] = 2.01-6.90) and helminthiasis (aHR = 2.27, 95% CI = 1.25-4.16) were higher than in the non-PI cohort. Patients with clonorchiasis had increased risks of gallstones without bile duct stones (aHR = 3.13, 95% CI = 1.54-6.39) and bile duct stones without gallstones (aHR = 4.75, 95% CI = 1.23-18.4). Patients with helminthiasis had an increased risk of gallstones without bile duct stones (aHR = 2.20, 95% CI = 1.11-4.33), but no higher risk of bile duct stones without gallstones was observed in helminthiasis (aHR = 2.22, 95% CI = 0.54-9.06), despite its high aHR. Neither clonorchiasis nor helminthiasis had an increased risk of concomitant gallstones and bile duct stones., Conclusion: Clonorchiasis is related to the development of not only gallstones, but also bile duct stones, and helminthiasis is only associated with the development of gallstones. However, cases of PI are rare in Taiwan, and this requires more international studies to clarify the association between helminthiasis and bile duct stones.