Clonorchis Sinensis Eggs Research Articles

A novel kit for enrichment of fecal helminth eggs.

We developed a new concentration kit, called the ParaEgg (PE), for easy detection trematode eggs from fecal samples in endemic areas of clonorchiasis and metagonimiasis in Korea. To create a standard of detection efficiency, 120 fecal samples were examined using the water-ether concentration method (WECM). The PE kit and Mini ParaSep (PS) kit were used to compare the detection sensitivity of 100 egg-positive and 20 egg-negative samples in WECM. Additionally, stool samples, which were intentionally spiked with 10, 20, and 30 Clonorchis sinensis eggs, were evaluated to assess the sensitivity in lowinfection cases. The PE and PS kits showed detection rates of 100% and 92%, respectively, from 100 egg-positive samples in WECM. Meanwhile, eggs were detected in 3 (PE) and 2 (PS) out of 20 egg-negative samples in WECM. The PE kit detected the highest number of eggs per gram of feces (727 on average), followed by the WECM (524) and PS kit (432). In fecal samples that were intentionally spiked with 10, 20, and 30 C. sinensis eggs, PE only detected eggs 2 out of 5 samples in 10 eggs spiked (40%), and the detection rates were 80% and 100%, respectively. The PE kit enabled a more accurate identification of trematode eggs because of the clearance of small fecal debris in the microscopic field. In conclusion, the PE kit is obviously helpful to detect and identify trematode eggs in stool examinations especially in endemic areas of clonorchiasis and metagonimiasis.

Comparative analysis of bile metabolic profile in patients with biliary obstruction complicated by Clonorchis sinensis infection.

Clonorchiasis is an important foodborne parasitic disease. However, eggs of Clonorchis sinensis (C. sinensis) cannot be detected in feces during biliary obstruction. Moreover, many diseases can cause biliary obstruction, such as gallstones, adenocarcinoma, cholangiocarcinoma and Ascaris lumbricoides infection. Therefore, it is of great significance to distinguish between patients of biliary obstruction and biliary obstruction with C. sinensis infection. A total of 48 biliary obstruction patients were enrolled, including 23 infected with C. sinensis (C. sinensis) (OB+C.s) and 25 non-infected subjects (OB). The bile samples were collected by endoscopic retrograde cholangiopancreatography and analyzed using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF MS). Additionally, multivariate statistical analysis methods were employed to identify differential metabolites. Next, bile amino acid levels were determined by targeted metabolomics analysis. A total of 146 and 132 significant metabolites were identified in electrospray ionization (ESI)+ and ESI- modes, respectively. The levels of amino acids (asparagine, glutamate, ornithine) and polyamines (spermidine and spermine) were significantly changed. Targeted analysis showed that the levels of amino acids (such as L-arginine, L-glutamine, L-lysine, L-propionic, and L-tyrosine) were lower in OB+C.s patients compared to those in OB patients. Marked metabolic pathways were involved in "Glutathione metabolism", "Caffeine metabolism", "Alanine, aspartate and glutamate metabolism", "Arginine and proline metabolism", "Purine metabolism", "Beta-Alanine metabolism", and "D-glutamine and D-glutamate metabolism". These results show that there were significant differences between OB+C.s and OB patients, especially in amino acids. The metabolic signature and perturbations in metabolic pathways may help to better distinguish OB+C.s and OB patients.

Traditional Lifestyle of the Indigenous Population of the Khabarovsk Krai as a Factor Determining the Spread of Helminth Infections

Introduction: Supporting health and wellbeing of indigenous small-numbered peoples of the North is one of the priorities of the regional policy in the Khabarovsk Krai. The centuries-long adaptation of the population to local conditions has created a unique system of the traditional use of natural resources and the most efficient forms and types of households. Despite all globalization processes, fishing, hunting and gathering remain essential elements of the lifestyle of northern indigenous peoples. Against the natural background, specific features of their behavior and nutrition create optimal conditions for helminth life cycles and intense transmission of endemic helminth infections. Objective: To assess effects of the traditional lifestyle of the indigenous population of the Khabarovsk Krai as a factor responsible for the spread of larval helminthiasis and endemic trematode infections. Materials and methods: In 2010–2020, questionnaire-based surveys were conducted and feces samples were collected from Nanai people in the villages of Dada, Sikachi-Alyan, Sinda, and Naikhin and from Udege people in the villages of Gvasyugi and Arsenyevo during expeditions. Results: Endemic trematode eggs were detected in stool samples from all the villages. Clonorchis sinensis eggs were detected in the samples from Dada, Sinda, and Sikachi-Alyan, while Nanophyetus salmincola schikhobalowi eggs were found in the dwellers of Arsenyevo and Gvasyugi. Metagonimus spp. eggs were also detected in isolated cases. The results of sero-epidemiological survey indicate a high frequency of contacts with Toxocara roundworms in all areas and with tapeworms of the Echinococcus type in the traditional villages located in forest areas. Conclusion: Original lifestyle and dietary habits of indigenous peoples in the Khabarovsk Krai pose a high risk of trematode infections (clonorchiasis, nanophietiasis, metagonimiasis) and larval helminthiasis (echinococcosis, toxocarosis). In combination with poor awareness of the population of appropriate preventive measures, this risk contributes to high incidence and prevalence rates of helminthiases in all surveyed national villages.

GENETIC ANALYSIS OF ANCIENT CLONORCHIS SINENSIS EGGS ATTAINED FROM GORYEONG MUMMY OF JOSEON DYNASTY PERIOD.

Although Clonorchis sinensis is a parasite that still infects many people in East Asia, its genetics remain largely unknown. We conducted ancient DNA analysis of C. sinensis eggs obtained from a Joseon period mummy newly discovered in South Korea. Clonorchis sinensis DNA was amplified for internal transcribed spacer 1, cytochrome c oxidase subunit 1, and NADH dehydrogenase subunit 2 and 5 genes. The results of BLAST/NCBI showed that the consensus sequences were 98.24 to 100% identical to the modern and ancient C. sinensis sequences reported from Korea, China, Japan, and other Asian countries. Our report helps to fill in the genetic profile of ancient C. sinensis strains that infected East Asian people hundreds of years ago.

A systematic study on 33 gallbladder stones resembling adult Clonorchis sinensis worms.

Limited studies provide direct evidence of Clonorchis sinensis adults in the early stage of gallbladder stone formation. Our current research systematically studied 33 gallbladder stones resembling adult worms and shed light on the definite connection of C. sinensis infection with concomitant cholelithiasis. A total of 33 gallbladder stones resembling adult C. sinensis worms were systematically analysed. Fourier transform infrared spectroscopy, scanning electron microscopy and X-ray energy spectrometry were used to analyse the composition and microstructure. Meanwhile, a histopathological examination of the stone was carried out. The 33 gallbladder stones resembling adult C. sinensis worms included nine calcium carbonate (CaCO3) stones, 12 bilirubinate stones and 12 mixed stones. Clonorchis sinensis eggs were found in 30 cases, including all CaCO3 and mixed stones. Parasite tissues were detected in 12 cases, which were mainly CaCO3 stones or bilirubinate-CaCO3 mixed stones. The outer layer of stones was wrapped with 12.88% calcium salt, as revealed by X-ray energy spectrometry, while surprisingly, many C. sinensis eggs were found in the inner part of these stones. Based on our current findings, we concluded that calcification and packaging occurred after C. sinensis adult entrance into the gallbladder, subsequently leading to the early formation of CaCO3 or bilirubinate-CaCO3 mixed gallbladder stones. This discovery highlights definite evidence for C. sinensis infection causing gallbladder stones.

Novel mechanism of hepatobiliary system damage and immunoglobulin G4 elevation caused by Clonorchis sinensis infection.

Clonorchis sinensis infection is still a major public health problem. It is estimated that more than 15 million people worldwide are infected, especially in Northeast China, Taiwan, South Korea, and North Vietnam. The detection of Clonorchis sinensis eggs in feces and bile is still the only gold standard for the diagnosis of Clonorchis sinensis infection, and new detection methods are needed to improve the detection rate. After Clonorchis sinensis invades the human body, it mainly parasitizes the hepatobiliary tract. Therefore, it is closely related to hepatobiliary diseases such as cholangitis, bile duct stones, liver fibrosis, and cholangiocarcinoma. The increase in immunoglobulin G4 (IgG4) caused by Clonorchis sinensis infection is rare and there are few reports about the relevant mechanism. It may be related to the inflammatory factors interleukin (IL)-4, IL-10, and IL-13 produced by human phagocytes, T cells, B cells, and other immune cells in the process of resisting the invasion of Clonorchis sinensis. However, this finding still needs further clarification and confirmation. This article reviews the epidemiology, clinical manifestations, serology, imaging, pathogenic mechanism, and control measures of Clonorchis sinensis infection to help establish the diagnostic process for Clonorchis sinensis. We report novel mechanisms of IgG4 elevation due to Clonorchis sinensis infection to provide more experience and a theoretical basis for clinical diagnosis and treatment of this infection.

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.

Value of discriminant analysis for screening of clonorchiasis sinensis

To build a discriminant function of clonorchiasis sinensis using stepwise discriminant analysis, so as to investigate the feasibility of discriminant analysis for clonorchiasis sinensis screening. Ten villages in Mulan County of Harbin City were sampled as the study sites using a cluster random sampling method. The fecal samples were collected from the permanent residents in the study sites and detected for Clonorchis sinensis eggs using the modified Kato-Katz smear technique. A questionnaire of clonorchiasis sinensis was designed. All data were entered into Epidata 3.1, and a discriminant analysis was performed using the software SPSS version 15.0. Variable were screened using the stepwise discriminant analysis, and the discriminant function was built using the Fisher's discriminant analysis method. The effectiveness of the discriminant function for clonorchiasis sinensis screening was evaluated by comparison with the modified Kato-Katz smear method. Eight variables with statistical significance were included to build the discriminant function, including chronic cholecystitis, cholangitis, gender, eating raw fish, abdominal distension, ethnicity, abdominal pain and age, and the correction rate of the discriminant function was 88.75% to identify clonorchiasis sinensis. The discriminant function of clonorchiasis sinensis can be rapidly and simply built based on the strong data processing and analysis capability of the SPSS software, which is rapid to screen clonorchiasis sinensis in Harbin City. Such a function has a high discriminant analysis capability, and provides insights into the establishment of rapid screening of clonorchiasis sinensis in other endemic areas.

Establishment and evaluation of the detection method of Cryptosporidium specific DNA fragment by recombinase aided isothermal amplification

To establish a recombinase-aided isothermal amplification (RAA) assay for detection of Cryptosporidium. Based on Cryptosporidium-specific 18S rRNA selected as the target gene to be detected, and the primer sequences and fluorescent probes designed using the software Amplfix, and a fluorescent RAA assay was established and optimized. The fluorescent RAA assay was performed to detect 18S RNA target sequence-contained recombinant plasmids at various copies, genomic DNA of Cryptosporidium oocysts at various concentrations, and genomic DNA extracted from various numbers of Cryptosporidium oocysts to assess the sensitivity of the assay, and to detect genomic DNA extracted from Cryptosporidium oocysts, Giardia lamblia cysts, Schistosoma japonicum eggs, Ascaris lumbricoides eggs, Clonorchis sinensis eggs, Salmonella and Shigella to determine the specificity of the assay. A fluorescent RAA assay was successfully established, which was effective to amplify the specific 18S RNA gene fragments of Cryptosporidium within 20 min at 39 ℃. The lowest limits of the fluorescent RAA assay were 102 copies/μL for detection of 18S RNA target sequence-contained recombinant plasmids at various copies, 1 pg/μL for detection of genomic DNA of Cryptosporidium oocysts at various concentrations, and one Cryptosporidium oocyst/μL for detection of genomic DNA extracted from various numbers of Cryptosporidium oocysts, and the fluorescent RAA assay was all negative for detection of genomic DNA from G. lamblia cysts, S. japonicum eggs, A. lumbricoides eggs, C. sinensis eggs, Salmonella and Shigella. A novel fluorescent RAA assay is successfully established, which is simple, rapid, sensitive and specific to detect genomic DNA of Cryptosporidium oocysts.

Retrospective study of pleural parasitic infestations: a practical diagnostic approach

BackgroundPleural parasitic infestation (PPI) is a disease prevalent in certain parts of the world. It is frequently misdiagnosed due to its lack of standardized diagnostic criteria. The purpose of this study was to evaluate the clinical characteristics of PPI patients and develop a practical diagnostic approach for PPI.MethodsA retrospective study was conducted by reviewing the medical records of 11 patients with PPI. A practical diagnostic approach was proposed based on the unique laboratory findings.ResultsAll patients demonstrated respiratory symptoms, including shortness of breath, cough, fever, chest pain, excessive sputum and hemoptysis. Leukocytosis (> 10,000/μL) and eosinophilia (> 500/μL) of peripheral blood were present in 45.5 and 36.4% patients, respectively. The mean concentrations of pleural effusion lactate dehydrogenase (LDH), adenosine deaminase (ADA), protein and carcinoembryonic antigen (CEA) were 338.2 U/L (range, 61–667 U/L), 11.6 U/L (range, 0.1–28.2 U/L), 43.7 g/dL (range, 21.9–88.1 g/dL), and 1.84 mg/mL (range, 0.28–4.8 mg/mL), respectively. The mean percentage of eosinophils in the pleural effusion was 19.5% (10.5–41%). Blood test was positive for parasite-specific IgG antibody in 9 patients, including 4 for Paragonimus westermani, 3 for Taenia solium, 1 for Clonorchis sinensis and 1 for Echinococcus granulosus. Eggs of Clonorchis sinensis were detected in the stool of two patients. Sparganum was found in the pleural effusion of one patient. Respiratory symptoms and abnormal appearances in pulmonary radiographic examination were disappeared in all patients who received anti-parasitic treatment.ConclusionsIn patients with unexplained pleural effusion, parasite-specific IgG antibody tests should be performed when pleural fluid testing shows eosinophilic pleural effusion. It is preferable to consider the diagnosis of PPI in clinical practice when serum parasite-specific IgG antibody test is positive.

Multiple food-borne trematodiases with profound systemic involvement: a case report and literature review

BackgroundFood-borne trematodiases are an important group of neglected global diseases. Affected patients in regions with low prevalence usually experience delayed diagnosis, especially when presenting with atypical clinical symptoms. Here, we presented a rare case of a Chinese patient infected with three food-borne trematodiases.Case presentationA 42-year-old man presented with diarrhea, lower extremity edema, and symptoms of cardiac dysfunction. He had a history of intermittent consumption of raw freshwater fishes for 6–7 years. Upon evaluation, he had eosinophilia, anemia, intrahepatic bile duct dilatation and a growing space-occupying lesion in the left atrium. The patient underwent a cardiac surgery which revealed an endocardial hematoma due to mechanical injuries. Imaging investigations also revealed intracranial and pulmonary lesions. A total of three trematodiases were diagnosed based upon microscopic stool examination, from which eggs of Clonorchis sinensis, Heterophyidae and Echinostomatidae were identified. Deposition of Clonorchis sinensis eggs was also observed from ileocecal squash slides. The patient was successfully treated with three cycles of praziquantel.ConclusionsFood-borne trematodiases may present with systemic involvement. Patients with dietary history of high risk or atypical ingestions should be evaluated for parasitic infection, even in non-endemic areas.

Investigation on epidemic status of clonorchiasis sinensis based on inpatients in Zhongshan City, China

Objective To understand the prevalence and epidemiological features of clonorchiasis sinensis based on inpatients in Zhongshan City. Methods All of the 23 town hospitals in Zhongshan City were selected as the surveillance hospitals in 2016. Among the inpatients from those hospitals, those lived in the city longer than 6 months were selected as the surveillance subjects. The stool specimens of the subjects were collected and examined by the modified Kato-Katz thick smear method for the eggs of Clonorchis sinensis. The crude infection rates of C. sinensis of the subjects were standardized according to the age and sex compositions of the population in Zhongshan City, 2016. Meanwhile, the infection rates of the subjects from different towns and those combined with related clinical diseases were analyzed. Results A total of 2 667 people were included in the surveillance, among which 219 were tested as positives, with a crude infection rate of 8.21%. For the positives, the male accounted for 81.74% (179/219), and their age ranged from 4 to 89 years with the mean value of (62.20±13.80) years. After standardization by age and sex composition, the infection rate was 3.62% for the whole population, the rates of the male and the female were 5.46% and 1.53%, respectively, and the former was 3.57 times as high as the latter. Both the infection rates of the male and the female rose as the age increasing (χ2male = 99.91, χ2female = 16.51, both P < 0.01). The standardized infection rates among the subjects in different towns ranged from 0 to 10.40%, and the rates in the north towns were higher than those in the south towns. The standardized rate was 9.22% for the subjects combined with gallstone, 3.40% for those not combined with gallstone, and the former was 2.71 times as high as that of the latter. Conclusion The people aged 30 years and above, with gallstone or living in towns with high C. sinensis infection rates are the key population for prevention and control of clonorchiasis in Zhongshan City.

Investigation of health knowledge and behaviors on clonorchiasis sinensis of urban residents in Yanqing District, Beijing City

Objective To understand the status of health knowledge and behaviors on clonorchiasis sinensis of urban residents in Yanqing District of Beijing City, China, so as to provide the targeted suggestions for further health education. Methods The communities/streets were selected as the investigation sites by using the stratified randomly sampling method. A total of 283 residents were randomly selected and investigated with the structural questionnaire designed by Beijing Center for Disease Control and Prevention. The fecal samples were collected and the eggs of Clonorchis sinensis were detected with Kato-Katz technique. Results All of the questionnaires were valid. The awareness rate about clonorchiasis was 25.09%. The rates of knowing clonorchiasis infection route, harm and prevention measures were 10.60%, 9.89% and 10.60% respectively. Totally 7.07% of the residents had the habits of eating raw or undercooked fresh water fish, and 2.47% of the residents had the behavior of eating sashimi. C. sinensis eggs were not found among the research objects. Conclusions The awareness rate about clonorchiasis is low among urban residents in Yanqing District. The focus of the future work is to extensively carry out the health education about clonorchiasis prevention and control and take relevant interventions.

A comparative study of mud‐like and coralliform calcium carbonate gallbladder stones

To gain insight to underlying mechanism of the formation of calcium carbonate (CaCO3 ) gallbladder stones, we did comparative study of stones with mud appearance and those with coralliform appearance. A total of 93 gallbladder stones with mud appearance and 50 stones with coralliform appearance were analyzed. The appearance, color, texture, and the detection of Clonorchis sinensis eggs by microscopic examination were compared between the two groups. Then, the material compositions of stones were analyzed using Fourier Transform Infrared spectroscopy and the spectrogram characteristics were compared. Moreover, microstructure characteristics of the two kinds of stones were observed and compared with Scanning Electron Microscopy. Mud-like gallbladder stones were mainly earthy yellow or brown with brittle or soft texture, while coralliform stones were mainly black with extremely hard texture, the differences between the two groups was significant (p < .05). The analytic results of FTIR spectroscopy showed that 95.7% (89/93) of the mud-like gallbladder stones were CaCO3 stones, and mainly aragonite; while all of the coralliform stones were CaCO3 stones, and mainly calcite (p < .05). Meanwhile, microscopic examination indicated that the detection rate of Clonorchis sinensis eggs in mud-like CaCO3 stones was lower than that in coralliform CaCO3 stones (p < .05), and that in aragonite CaCO3 stones was lower than that in calcite CaCO3 stones(p < .05). Mud-like CaCO3 stones mainly happened to patients with cystic duct obstruction. Clonorchis sinensis infection was mainly associated with coralliform (calcite) CaCO3 stones. Cystic duct obstruction was mainly associated with mud-like (aragonite) CaCO3 stones.

Association between Clonorchis sinensis infection and the subtypes of mixed gallbladder stones

Objective A systematic study to classify mixed gallbladder stones to determine their relationship with Clonorchis sinensis infection. Methods 349 consecutive patients with mixed gallbladder stones were enrolled into this study and their gallstones were studied. The material composition of gallbladder stones was analyzed using Fourier Transform Infrared spectroscopy, and the distribution and microstructure of the material components were studied using Scanning Electron Microscopy. The composition and distribution of the elements were analyzed by an X-ray energy spectrometer. Gallbladder stones were classified accordingly, and the stones were then grinded for microscopic examination. The stones were divided into two groups based on the results of detection of clonorchis sinensis eggs. Patients were also divided into two groups according to the patients' region (high-incidence and low-incidence regions of Clonorchis sinensis infection). The diversity of the subtypes of mixed gallbladder stones was compared. Results 14 subtypes were found, including Bilirubinate-calcium carbonate mixed stones (n=134), Cholesterol-bilirubinate mixed stones (n=87), Bilirubinate-phosphate mixed stones (n=39), Cholesterol-calcium carbonate mixed stones (n=30), Cholesterol-bilirubinate -calcium carbonate mixed stones (n=15), etc. Bilirubinate-calcium carbo-nate mixed stones were the main subtype of egg-positive mixed gallbladder stones and the patients were from high-incidence regions. Cholesterol-bilirubinate mixed stones were the main subtype of egg-negative mixed gallbladder stones and patients were from low-incidence regions. The differences of the subtypes constituent between the high-incidence and low-incidence regions groups were significant (P<0.05), and between the egg-positive and egg-negative groups were also significant (P<0.05). Conclusions The most common subtypes of mixed gallbladder stones were bilirubinate-calcium carbonate mixed stones, while the second was cholesterol-bilirubinate mixed stones. Clonorchis sinensis infection was mainly associated with Bilirubinate-calcium carbonate mixed stones. Key words: Gallbladder Stones; Mixed Stone; Fourier transform infrared spectroscopy; Scanning electronic microscopy; X-ray photoeletron spectroscopy

A Case of Malignant Biliary Stricture Mimicking Benign Stricture coincided with &amp;lt;italic&amp;gt;Clonorchis sinensis&amp;lt;/italic&amp;gt;

Accurate and early diagnosis of indeterminate bile duct stricture is difficult. There are numerous cases suggesting similarity between benign tumors and malignancy. Therefore, meticulous evaluation with endoscopic retrograde cholangiopancreatography (ERCP), endoscopic ultrasound and computed tomography (CT) is necessary. A 50 year-old male presented with painless jaundice. Abdominal CT scan showed distal biliary stricture without definite pancreatic mass. Repeated brush cytology and endobiliary biopsy during ERCP did not reveal malignancy except for eggs of Clonorchis sinensis . The patient declined surgical resection without definite evidence of malignancy. Abdominal CT scan one month later showed progressive parenchymal atrophy and pancreatic duct dilatation. The patient underwent pylorus preserving pancreatoduodenectomy. Pathology revealed pancreatic adenocarcinoma in the head portion. Since accurate preoperative diagnosis of malignant biliary obstruction can be evasive, patients with biliary stricture should undergo evaluation with high index of suspicion. Korean J Pancreatobiliary 2015;20(1):51-56

V-shaped pits in regions of ancient Baekje kingdom paleoparasitologically confirmed as likely human-waste reservoirs.

In a paleo-parasitological analysis of soil samples obtained from V-shaped pits dating to the ancient Baekje period in Korean history, we discovered Ascaris lumbricoides, Trichuris trichiura, and Clonorchis sinensis eggs. In light of the samples' seriously contaminated state, the V-shaped pits might have served as toilets, cesspits, or dung heaps. For a long period of time, researchers scouring archaeological sites in Korea have had difficulties locating such structures. In this context then, the present report is unique because similar kind of the ancient ruins must become an ideal resource for successful sampling in our forthcoming paleoparasitological studies.

The Changing Pattern of Parasitic Infection Among Korean Populations by Paleoparasitological Study of Joseon Dynasty Mummies

In the coprolites of 4 recently discovered Joseon mummies of Korea, we found Ascaris lumbricoides , Trichuris trichiura , Metagonimus yokogawai , Paragonimus westermani , and Clonorchis sinensis eggs. The current finding was compared with previous paleoparasitological data, and with recent national survey data from Korea. For A. lumbricoides and T. trichiura , similar patterns of infection prevalence were observed between the national survey of 1961 and our current Joseon data. Some of the trematode species (C. sinenesis and P. westermani) showed much higher infection prevalences among the Joseon Koreans than among their 1960s descendants. The present results indicate that the decrease in trematode infection rates might have begun earlier than was the case for nematode infection.

Development of a polymerase chain reaction applicable to rapid and sensitive detection of Clonorchis sinensis eggs in human stool samples

Microscopic examination of eggs of parasitic helminths in stool samples has been the most widely used classical diagnostic method for infections, but tiny and low numbers of eggs in stool samples often hamper diagnosis of helminthic infections with classical microscopic examination. Moreover, it is also difficult to differentiate parasite eggs by the classical method, if they have similar morphological characteristics. In this study, we developed a rapid and sensitive polymerase chain reaction (PCR)-based molecular diagnostic method for detection of Clonorchis sinensis eggs in stool samples. Nine primers were designed based on the long-terminal repeat (LTR) of C. sinensis retrotransposon1 (CsRn1) gene, and seven PCR primer sets were paired. Polymerase chain reaction with each primer pair produced specific amplicons for C. sinensis, but not for other trematodes including Metagonimus yokogawai and Paragonimus westermani. Particularly, three primer sets were able to detect 10 C. sinensis eggs and were applicable to amplify specific amplicons from DNA samples purified from stool of C. sinensis-infected patients. This PCR method could be useful for diagnosis of C. sinensis infections in human stool samples with a high level of specificity and sensitivity.

Production and Deformation of Clonorchis sinensis Eggs during In Vitro Maintenance

Clonorchis sinensis is a carcinogenic human liver fluke. The present study monitored eggs produced by long-term maintained adult worms of C. sinensis to confirm their egg productivity in vitro. The worms from infected rabbits were incubated in vitro in 1× Locke’s solution and broth media (RPMI-1640, DMEM and IMDM). Numbers of expelled eggs were counted sequentially and their morphological changes were monitored by microscopy after 1, 30, 60, and 90 days of cultivation. On the 1–3 days of cultivation, the eggs counted maximum 4,756±202 eggs/worm/day in IMDM medium. The number of eggs gradually decreased less than 1,000 at 7–14 days and below 100 at 21days but continued to pass eggs after 56 days in all media. Length of the eggs were reduced about 1 µm at 30 days, and the length/width ratio was maintained around 1.8 at 30 days but decreased to 1.7 at 60 days and 1.5 at 90 days. Faust-Meleney index (FMI) decreased as the cultivation duration increased and lowest FMI (5662.9±974.7) observed in IMDM media at day 90 (P = 0.001). Microscopic findings of the eggs recognized the miracidium in most of eggs at 60 days but not in those at 90 days. Instead, the eggs contained dark granules or vacuoles in the deformed shell at 90 days. Scanning electron microscopy revealed partial loss of wrinkles on the deformed egg surface and prominent abopercular knob. Eggs viability decreased as the cultivation progressed and showed significant positive correlation with FMI and length/width ratio. In conclusion, the cultivated worms pass only the eggs which are preformed in their uterus before cultivation. One gravid C. sinensis contains about 37,000 eggs in its uterus and produces about 4,000 eggs every day. The deformed eggs with FMI less than 7,000 and length/width ratio lower than 1.7 are non-viable.