Clonorchiasis in Patients with Biliary and Pancreatic Diseases: Diagnosis and Risk Factors.

BACKGROUND: Many epidemiological studies have investigated the risk factors for clonorchiasis, but endoscopic findings of this disease in endoscopic retrograde cholangiopancreatography (ERCP) have not been well characterized. In this study, we evaluated clonorchiasis in ERCP in patients with biliary and pancreatic diseases. METHODS: This was a retrospective two-center study in hospitalized patients who received ERCP between January 2012 and October 2018. All patients were divided into clonorchiasis and nonclonorchiasis groups. Data were analyzed using univariate analysis and multivariate analyses. RESULTS: A total of 1119 patients were included, and clonorchiasis was diagnosed in 19.2% patients. Detection of Clonorchis sinensis eggs in bile samples was higher than that in fecal samples (85.9% vs. 58.7%; P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249. CONCLUSIONS: The detection of C. sinensis eggs was significantly higher in bile than in fecal samples; thus, bile samples represent a preferable sample for the diagnosis of clonorchiasis in patients with biliary obstruction. We found that male, age ≤ 60 years old, and CBD diameter < 12 mm were independent risk factors for clonorchiasis, while papilla fistula was a protective factor.C. sinensis eggs was significantly higher in bile than in fecal samples; thus, bile samples represent a preferable sample for the diagnosis of clonorchiasis in patients with biliary obstruction. We found that male, age ≤ 60 years old, and CBD diameter < 12 mm were independent risk factors for clonorchiasis, while papilla fistula was a protective factor.

Expanded complement of Niemann-Pick type C2-like protein genes in Clonorchis sinensis suggests functions beyond sterol binding and transport.

BACKGROUND: The parasitic flatworm Clonorchis sinensis inhabits the biliary tree of humans and other piscivorous mammals. This parasite can survive and thrive in the bile duct, despite exposure to bile constituents and host immune attack. Although the precise biological mechanisms underlying this adaptation are unknown, previous work indicated that Niemann-pick type C2 (NPC2)-like sterol-binding proteins might be integral in the host-parasite interplay. Expansions of this family in some invertebrates, such as arthropods, have shown functional diversification, including novel forms of chemoreception. Thus, here we curated the NPC2-like protein gene complement in C. sinensis, and predicted their conserved and/or divergent functional roles. METHODS: We used an established comparative genomic-bioinformatic approach to curate NPC2-like proteins encoded in published genomes of Korean and Chinese isolates of C. sinensis. Protein sequence and structural homology, presence of conserved domains and phylogeny were used to group and functionally classify NPC2-like proteins. Furthermore, transcription levels of NPC2-like protein-encoding genes were explored in different developmental stages and tissues. RESULTS: Totals of 35 and 32 C. sinensis NPC2-like proteins were predicted to be encoded in the genomes of the Korean and Chinese isolates, respectively. Overall, these proteins had low sequence homology and high variability of sequence alignment coverage when compared with curated NPC2s. Most C. sinensis proteins were predicted to retain a conserved ML domain and a conserved fold conformation, with a large cavity within the protein. Only one protein sequence retained the conserved amino acid residues required in bovine NPC2 to bind cholesterol. Non-canonical C. sinensis NPC2-like protein-coding domains clustered into four distinct phylogenetic groups with members of a group frequently encoded on the same genome scaffolds. Interestingly, NPC2-like protein-encoding genes were predicted to be variably transcribed in different developmental stages and adult tissues, with most being transcribed in the metacercarial stage. CONCLUSIONS: The results of the present investigation confirms an expansion of NPC2-like proteins in C. sinensis, suggesting a diverse array of functions beyond sterol binding and transport. Functional explorations of this protein family should elucidate the mechanisms enabling the establishment and survival of C. sinensis and related flukes in the biliary systems of mammalian hosts.

Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways.

BACKGROUND: MF6p/host defense molecules (HDMs) are a broad family of small proteins secreted by helminth parasites. Although the physiological role of MF6p/HDMs in trematode parasites is not fully understood, their potential biological function in maintaining heme homeostasis and modulating host immune response has been proposed. METHODS: A gene encoding the MF6p/HDM of Clonorchis sinensis (CsMF6p/HDM) was cloned. Recombinant CsMF6p/HDM (rCsMF6p/HDM) was expressed in Escherichia coli. The biochemical and immunological properties of rCsMF6/HDM were analyzed. CsMF6p/HDM induced pro-inflammatory response in RAW 264.7 cells was analyzed by cytokine array assay, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay. The structural feature of CsMF6p/HDM was analyzed by three-dimensional modeling and molecular docking simulations. RESULTS: The CsMF6p/HDM shares a high level of amino acid sequence similarity with orthologs from other trematodes and is expressed in diverse developmental stages of the parasite. The rCsMF6p/HDM bound to bacteria-derived lipopolysaccharide (LPS), without effectively neutralizing LPS-induced inflammatory response in RAW 264.7 macrophage cells. Rather, the rCsMF6p/HDM induced pro-inflammatory immune response, which is characterized by the expression of TNF-α and IL-6, in RAW 264.7 cells. The rCsMF6p/HDM-induced pro-inflammatory immune response was regulated by JNK and p38 MAPKs, and was effectively down-regulated via inhibition of NF-κB. The structural analysis of CsMF6p/HDM and the docking simulation with LPS suggested insufficient capture of LPS by CsMF6p/HDM, which suggested that rCsMF6p/HDM could not effectively neutralize LPS-induced inflammatory response in RAW 264.7 cells. CONCLUSIONS: Although rCsMF6p/HDM binds to LPS, the binding affinity may not be sufficient to maintain a stable complex of rCsMF6p/HDM and LPS. Moreover, the rCsMF6p/HDM-induced pro-inflammatory response is characterized by the release of IL-6 and TNF-α in RAW 264.7 macrophage cells. The pro-inflammatory response induced by rCsMF6p/HDM is mediated via NF-κB-dependent MAPK signaling pathway. These results collectively suggest that CsMF6p/HDM mediates C. sinensis-induced inflammation cascades that eventually lead to hepatobiliary diseases.

A co-infection case report of Taenia saginata in a patient with subclinical clonorchiasis confirmed by the combination of diagnostic tools.

BACKGROUND: Clonorchiasis is the common parasitic infection in the general population of the Republic of Korea, however, taeniasis is scarcely reported recently. Here, we describe a case of co-infection with the cestode T. saginata in a patient with subclinical clonorchiasis diagnosed by a combination of diagnostic tools in Korea. CASE PRESENTATION: A 56-year-old man visited the hospital having passed proglottids in his stool for the past two months and brought a stool sample with segments to our hospital. He had no abdominal symptoms, such as nausea, vomiting, abdominal pain, diarrhea, or constipation. He used to consume raw beef and fish frequently. We could not find evidence of gravid proglottids which contain fully developed uteri filled with ova or branched uterine structures, within the submitted sample. To identify the tapeworm species, we carried out molecular analyses on the proglottids. The cox1 and ef1a sequences had a 100% match with those of T. saginata and differed from the sequences of the other Taenia species. Upon examination of stool samples fixed by formalin-ether concentration method, no Taenia species ova were observed in 10 slides. Instead, C. sinensis ova were observed, despite the level of IgG specific to C. sinensis being within the normal range. The patient was treated with praziquantel (25 mg/kg, three times a day) for 3 days, and subsequently C. sinensis ova were not found in his stool. CONCLUSION: Our case indicates that a combination of morphological, serological, and molecular diagnostic tools should be used for the accurate diagnosis of subclinical parasitic infections.

Rare cause of jaundice in a post liver transplant patient.

A hepatitis B virus carrier suffering from acute flare of chronic hepatitis B infection underwent deceased-donor liver transplantation. He was put on the immunosuppressive agent tacrolimus. On routine follow-up, he was found to have abnormal liver function. Computed tomography scan of the abdomen did not show any dilatation of the biliary system. Liver biopsy showed scattered microabscesses, and a microgranuloma was detected. Endoscopic retrograde cholangiography was performed and a biliary anastomotic stricture (BAS) was noted. In addition, the Chinese liver fluke, Clonorchis sinensis, was discovered. Balloon dilatation and stenting were performed. The patient was given a course of praziquantel. His liver function improved and normalized. We present the case of a liver transplant recipient with cholangitis caused by C. sinensis infestation and infection and biliary obstruction resulting from BAS.

Correlation between sonographic findings and infection intensity in clonorchiasis.

This prospective study was performed to evaluate the diagnostic usefulness of sonography in terms of the intensity of Clonorchis sinensis infection. Total 1,384 residents were subjected to this study at an endemic area in China, in which a clonorchiasis control program had been performed. History taking, fecal examination, and sonography were performed, and 87.8% of the subjects were found to be egg positive. Sonography showed intrahepatic bile duct dilatation (IHDD) in 68.4% of the subjects, increased periductal echogenicity (IPDE) in 24.5%, and gallbladder sludge (GBS) in 20.8%. Moreover, the grades of these sonographic findings were higher in subjects with heavier infections as determined by egg counts. IHDD was found to be the most accurate sonographic indicator of any infection degree (76.3), whereas IPDE and GBS were useful only in cases of heavy infection. Subjective symptoms were found to be only poorly correlated with sonographic findings or infection intensity.

Clinical and epidemiological features of patients with clonorchiasis.

AIM: To study the clinical and epidemiological features of patients with clonorchiasis so as to provide scientific evidences for the diagnosis and prevention of clonorchiasis. METHODS: Stools from 282 subjects suspected of having clonorchiasis were examined for helminth eggs with modified Kato's thick smear and sedimentation methods, and their sera were tested for HAV-DNA, HBV-DNA, HCV-RNA, HDV-RNA and HEV-RNA with polymerase chain reaction (PCR). Clinical symptoms of patients with clonorchiasis only were analyzed, and their blood samples were tested for circulating antigen (CAg) with Dot-ELISA, eosinophilic granulocyte count, and alanine aminotransferase (ALT). Meanwhile, they were asked to provide data of occupation, eating habit, hygienic habit and knowledge of clonorchiasis. In addition, the ecosystem of the environment in epidemic areas was surveyed. RESULTS: Among the 282 patients, 61 (21.43%) were infected with clonorchis sinensis only, 97 (34.64%) were co-infected with clonorchis sinensis and other pathogens, 92 (32.86%) were infected with hepatitis virus only and 31 (11.07%) neither with clonorchis sinensis nor hepatitis virus. Among the 61 patients with clonorchiasis only, there were 14 (22.95%) subjects with discomfort over hepatic region or epigasfrium, 12 (19.67%) with general malaise or discomfort and inertia in total body, 6 (9.84%) with anorexia, indigestion and nausea, 4 (6.56%) with fever, dizziness and headache (6.56%), and 25 (40.98%) without any symptoms; sixty one (100%) with CAg (+), 98.33% (59/60) with eosinophilic granulocytes increased and 65.00% (39/60) with ALT increased. B-mode ultrasonography revealed 61 cases with dilated and thickened walls of intrahepatic bile duct, and blurred patchy echo acoustic image in liver. Twenty-six cases had stones in the bile duct, 39 cases had slightly enlarged liver with diffuse coarse spots in liver parenchyma. Twenty cases had enlarged gallbladder with thickened coarse wall and image of floating plagues, 9 cases had slightly enlarged spleen. By analysis of epidemiological data, we found that the ecologic environment was favorable for the epidemiology of clonorchiasis. Most patients with clonorchiasis were lack of knowledge about the disease. Their living environment, hygienic habits, eating habits and their occupations were the related factors that caused the prevalence of the disease. CONCLUSION: The clinical symptoms of clonorchiasis are non-specific, and the main evidences for diagnosis of clonorchiasis should be provided by etiologic examination, B-mode ultrasonography and clinical history. The infection of clonorchis sinensis is related to occupations, bad eating habits and lack of knowledge about prevention of the disease.

Biliary liver flukes (Opisthorchiasis and Clonorchiasis) in immigrants in the United States: often subtle and diagnosed years after arrival.

BACKGROUND: Biliary liver flukes are extremely common parasites in some regions of the world where consumption of raw fresh-water fish is a cultural practice. Clonorchis sinensis and Opisthorchis spp. are the major biliary liver flukes associated with human disease. Some of these parasites are highly associated with cholangiocarcinoma. It was noted that several cases presenting at our center had been in the US for prolonged periods. This prompted us to retrospectively review cases of liver fluke infection diagnosed at our facility. METHODS: All cases of biliary liver fluke infection over a 6-year period were retrospectively investigated at a clinic serving international patients, to determine the prevalence and risk factors for infection in a nonendemic area. Cases were identified through review of stool ova and parasite (O&P) records maintained at Regions Hospital/HealthPartners microbiology laboratory. All O&P samples positive for Opisthorchis spp. or Clonorchis sinensis were included in the review. RESULTS: Seventeen cases were identified during the study period. Our center performed approximately 1,800 stool O&P examinations per year on approximately 1,100 individuals per year during the study period. Biliary liver flukes were uncommon, accounting for 1.3% of individuals infected with organisms considered to be potentially pathogenic. Infections were predominantly found in migrants from Laos, Cambodia and Thailand. Cases were also identified in migrants from the former Soviet Union and South America. It is of note that 25% of patients were detected after 5 years of residence in the US. Often, the only clinical clue to infection was a mild absolute eosinophilia (500 to 1000 microL). CONCLUSIONS: Although biliary liver fluke is an infrequent cause of infection in immigrants to the US, because of the potential long-term consequences of chronic infection, educational information highlighting routes of infection and the fact that asymptomatic infection is common and may eventually lead to cancer should be aimed at high-risk community members and those who have been visiting friends and relatives in endemic areas.

Infecçäo por Clonorchis sinensis em imigrantes asiáticos no Brasil: tratamento com praziquantel / Clonorchis sinensis infections in Asian immigrants in Brazil: treatment with praziquantel

Estudaram-se quinze pacientes com infecçäo assintomática por Clonorchis sinensis, revelada através de exame parasitológico de fezes. Todos eram de origem asiática e procuraram o Laboratório Central do Instituto Adolfo Lutz para se submeterem a exames laboratoriais necessários a regularizaçäo de sua situaçäo, face a nova legislaçäo sobre imigrantes. Eram todos indivíduos adultos, seis pertencendo ao sexo feminino e nove ao masculino. Os quinze pacientes comn clonorquíase foram internados no Hospital das Clínicas da FMUSP e tratados com Praziquantel, na dosagem de 60 mg/Kg de peso corporal, dividida em duas tomadas. Foram realizados exames coprológicos quantitativos (método de Kato-Katz), antes do tratamento específico e no 15§, 30§ e 60§ dias após a terapêutica. Na última avaliaçäo (60§ dias após terapêutica), em nove pacientes (60,0%) näo se encontraram ovos do treamatódeo nas fezes e nos seis (40,0%), que continuavam eliminando ovos, notou-se reduçäo na quantidade eliminada (superior a 90% em cinco e a 30% no paciente restante). Os pacientes foram também submetidos a exames subsidiários, para avaliaçäo do estado geral e funçäo hepática, antes da administraçäo de Praziquantel e, posteriormente, no seguimento ambulatorial. A medicaçäo foi relativamente bem tolerada pelos pacientes, verificando-se a ocorrência de efeitos colaterais representados por náuseas e vômitos (dois casos), vertigens e tonturas (dois casos), epigastralgia (dois casos) e diarréia no 3§ dia após tratamento (um caso)