Clonorchiasis: an update.

Clonorchis sinensis, the Chinese or oriental liver fluke, is an important human parasite and is widely distributed in southern Korea, China (including Taiwan), Japan, northern Vietnam and the far eastern part of Russia. Clonorchiasis occurs in all parts of the world where there are Asian immigrants from endemic areas. The human and animal reservoir hosts (dogs, pigs, cats and rats) acquire the infection from the ingestion of raw fish containing infectious metacercariae. The first intermediate snail hosts are mainly species of Parafossarulus and Bithynia. Numerous species of freshwater fish serve as the second intermediate hosts of C. sinensis. Extensive studies of clonorchiasis during several decades in Japan, Korea, China and other countries have shown much progress in proving its morphological features including ultrastructure, biology, pathogenesis, epidemiology, clinical manifestations and chemotherapy. The present review deals with mainly current results obtained on the epidemiological, pathological and clinical aspects, as well as control measures in endemic areas. As for the complications of clonorchiasis, formation of calculi in the intrahepatic biliary passages is one of the most characteristic pathological features. It is sometimes accompanied by suppurative cholangitis, cholecystitis, cholangiohepatitis and ultimately can cause cholangiocarcinoma. Experimental results on the relationship to the occurrence of cholangiocarcinoma are presented. Clinical diagnosis by radiological findings including cholangiography, sonography and computerized tomography as well as magnetic resonance imaging for biliary or pancreatic ducts are outlined. Current studies on immunology and molecular biology of C. sinensis were introduced. Praziquantel is the drug of choice for clonorchiasis. The most effective regimen is 25 mg kg(-1) three times daily (total dose, 75 mg kg(-1)) administered orally at 5- to 6-h intervals over a single day. Prevention and control measures are also discussed.

Mixed infections with Opisthorchis viverrini and intestinal flukes in residents of Vientiane Municipality and Saravane Province in Laos.

Faecal examinations for helminth eggs were performed on 1869 people from two riverside localities, Vientiane Municipality and Saravane Province, along the Mekong River, Laos. To obtain adult flukes, 42 people positive for small trematode eggs (Opisthorchis viverrini, heterophyid, or lecithodendriid eggs) were treated with a 20-30 mg kg(-1) single dose of praziquantel and purged. Diarrhoeic stools were then collected from 36 people (18 in each area) and searched for helminth parasites using stereomicroscopes. Faecal examinations revealed positive rates for small trematode eggs of 53.3% and 70.8% (average 65.2%) in Vientiane and Saravane Province, respectively. Infections with O. viverrini and six species of intestinal flukes were found, namely, Haplorchis taichui, H. pumilio, H. yokogawai, Centrocestus caninus, Prosthodendrium molenkampi, and Phaneropsolus bonnei. The total number of flukes collected and the proportion of fluke species recovered were markedly different in the two localities; in Vientiane, 1041 O. viverrini (57.8 per person) and 615 others (34.2 per person), whereas in Saravane, 395 O. viverrini (21.9 per person) and 155207 others (8622.6 per person). Five people from Saravane harboured no O. viverrini but numerous heterophyid and/or lecithodendriid flukes. The results indicate that O. viverrini and several species of heterophyid and lecithodendriid flukes are endemic in these two riverside localities, and suggest that the intensity of infection and the relative proportion of fluke species vary by locality along the Mekong River basin.

Infectivity and pathological changes in murine clonorchiasis: comparison in immunocompetent and immunodeficient mice.

The main complications of clonorchiasis are periportal inflammation, biliary hyperplasia, periductal fibrosis, and subsequently the development of biliary tumors in the liver. This study was undertaken to compare the infectivity and histopathologic changes between in immunocompetent FVB/NJ and BALB/cA strains, and immunodeficient severe combined immunodeficient (SCID) and athymic nude mice after the metacercariae of Clonorchis (C.) sinensis were infected. The experiment showed that C. sinensis was very infective in all strains studies, but the status of worm development, infectivity, recovery rate, and morphological changes of livers were very different in each strain. FVB/NJ mice showed more worm recovery than any other strain. Histopathologically the liver of FVB/NJ mice at 4 weeks postinfection showed marked cystic and fibrotic changes, in which C. sinensis was fully developed with ovum production, severe infiltration of inflammatory cells, mostly eosinophils, and high degrees of biliary hyperplasia. In SCID and nude mice, there were few foci of inflammatory cells even at 8 weeks postinfection in periportal areas of the liver, associated with no development into adult worm with ovum production. Fibrosis occurring at 4 weeks postinfection was highly correlated with inflammatory infiltration when each strain was compared. We suggest that massive infiltration of eosinophil and plasma cells caused by the infection might initiate cystic formation and fibrosis. These data demonstrate that the infection of C. sinensis might be related to pathologic consequences of inflammatory cell infiltration, cystic formation and fibrosis which might play a role in the defense mechanism against the parasitism in the liver of each strain. The FVB/NJ mouse model might be very helpful in elucidating the mechanism for human clonorchiasis.

A nationwide survey of the prevalence of human Gymnophalloides seoi infection on western and southern coastal islands in the Republic of Korea.

A nationwide survey was performed to know the distribution and prevalence of human Gymnophalloides seoi infection on western and southern coastal islands in the Republic of Korea. A total of 4,178 fecal specimens were collected from residents on 45 (24 western and 21 southern) islands, and examined by Kato-Katz and formalin-ether sedimentation techniques. Eggs of G. seoi were detected from 160 (3.8%) people living on 22 (13 western and 9 southern) islands. The prevalence varied by the location of islands; higher on western islands than on southern islands. The highest prevalence was found on Amtaedo (25.3%), followed by Cheungdo (25.0%), and Anchwado (20.9%) (Shinan-gun). A little lower prevalence was observed on Munyodo (13.3%), Shinshido (12.9%), and Sonyudo (10.3%) (Kunsan-shi). Of the remaining islands, the regions showing the prevalence greater than 5% included Kohado, Dallido (Mokpo-shi), Pyeongildo, Kogumdo (Wando-gun), and Keogumdo (Kohung-gun). A strong age predilection was noted (p < 0.05); 95% of the infected people were over 40 years old. Females showed a little higher prevalence than males. The results indicate that human G. seoi infection is more widely distributed than previously considered. Nine of 11 islands (excluding the 2 known areas Munyodo and Sunyudo) that showed greater prevalence than 5% are regarded as new endemic foci of G. seoi.

Epidemiological understanding of Taenia tapeworm infections with special reference to Taenia asiatica in Korea.

In endemic areas of Taenia tapeworms in Korea, most of the reports showed that T. saginata was dominant over T. solium, but eating pigs is the dominant habit over eating cattle. Why do they have more T. saginata despite lower consumption of beef? This problem actually has long been recognized but until recently there has been no intensive trial to give a scientific explanation on this epidemiological enigma. By summing up the data published between the years 1963 and 1999, the ratio of armed versus unarmed tapeworms in humans was estimated at approximately 1:5. The ratio of pig-eaters versus cattle-eaters, however, was approximately 5:1. This inconsistency could be explained with the recently described T. asiatica, which infects humans through the eating of pig's viscera. We re-evaluate the importance of the consumption of visceral organ of pigs. leading us to an improved epidemiological understanding of the T. asiatica infection together with co-existing T. saginata and T. solium in Korea.

Control of clonorchiasis by repeated treatments with praziquantel.

The present study aimed to evaluate control efficacy of clonorchiasis by two schemes of repeated treatment with praziquantel at two endemic villages in China. Residents of one village at Guangxi Autonomous Region were treated and examined 6-monthly and of another at Liaoning Province 12-monthly. In residents that took 25 mg/kg x 3 (total 75 mg/kg) of praziquantel every 6 months for one year the egg positive rate showed a significant drop from 69.0% to 17.1%. In contrast, a group of same praziquantel medication once showed a slight marginal decrease in the egg rate from 18.9% to 12.2% after one year. Of 39 subjects examined 3 times, 56.4% were cured, 7.7% persistently positive, one (2.6%) reinfected after cure or newly infected, but 25.6% were persistently negative. The present finding suggests that 6-monthly medication with 75 mg/kg of praziquantel should effectively lower the prevalence but incomplete for control of clonorchiasis in heavy endemic areas.

Genotyping of Giardia lamblia isolates from humans in China and Korea using ribosomal DNA Sequences.

Genetic characterization of a total of 15 Giardia lamblia isolates, 8 from Anhui Province, China (all from purified cysts) and 7 from Seoul, Korea (2 from axenic cultures and 5 from purified cysts), was performed by polymerase chain reaction amplification and sequencing of a 295-bp region near the 5' end of the small subunit ribosomal DNA (eukaryotic 16S rDNA). Phylogenetic analyses were subsequently conducted using sequence data obtained in this study, as well as sequences published from other Giardia isolates. The maximum parsimony method revealed that G. lamblia isolates from humans in China and Korea are divided into 2 major lineages, assemblages A and B. All 7 Korean isolates were grouped into assemblage A, whereas 4 Chinese isolates were grouped into assemblage A and 4 into assemblage B. Two Giardia microti isolates and 2 dog-derived Giardia isolates also grouped into assemblage B, whereas Giardia ardeae and Giardia muris were unique.

Chemotherapeutic efficacy of praziquantel in rats with protective immunity to Clonorchis sinensis infection.

OBJECTIVE: To study the synergic effect of praziquantel (PZQ) and host acquired immunity on Clonorchis sinensis. METHODS: Acquired immunity to C. sinensis was induced by immunization with crude adult worm antigen (AW Ag) and excretory-secretory antigen (ES Ag) or infection with C. sinensis metacercariae. The effect was assessed by the worm reduction rate compared with the control groups after challenge infection with 50 metacercariae and treated orally with a subcurative dose of praziquantel (50 mg/kg). Significant test was performed by analysis of variance (ANOVA) and Npar1 way Kruskal-Wallis test. All calculations were performed by PC-SAS system. RESULTS: 1. PZQ was more effective against C. sinensis larvae than against adult worms in the control (P < 0.001), ES Ag (P < 0.01) or crude AW Ag immunization group (P < 0.001). 2. As compared with the control, the worm reduction rate after challenge infection was significantly higher (P < 0.001) in ES Ag immunized group (35.6%) and metacercaria infection group (97.5%) and less in crude AW Ag group (23.4%). The PZQ efficacy was significantly enhanced in ES Ag immunized group. CONCLUSION: The efficacy of PZQ against C. sinensis could be synergically enhanced in rats by inducing host acquired immunity.

Neoplagioporus zacconis (Trematoda: Opecoelidae) from the intestine of the pale chub, Zacco platypus, in Korea.

Neoplagioporus zacconis (Yamaguti, 1934) (Trematoda: Opecoelidae) was found from the intestine of the pale chub, Zacco platypus, for the first time in Korea. The worms were characterized morphologically by oval body shape, bipartited seminal vesicle, sinistrally located genital pore, and lobed ovary, and distinguished from the two other species of Neoplagioporous by body shape and distribution of vitelline follicles. The morphological characteristics, except the ovary, well corresponded to those of the previous descriptions. The morphological difference of the ovary was considered as a character of geographical phenotypic variation.

Heterogeneity of the "oval-cell" response in the hamster liver during cholangiocarcinogenesis following Clonorchis sinensis infection and dimethylnitrosamine treatment.

BACKGROUND: Small intraportal "oval" cells which appear in the livers of humans and experimental animals after liver injury, are suspected to be early progenitor cells for both hepatocytes and bile duct cells, as well as cells of origin of hepatocellular and cholangiocellular carcinomas. METHODS: The origin and fate of small "oval" cells expressing different immunohistologic phenotypes and ultrastructural appearance were examined in livers of Syrian hamsters during cholangiocarcinogenesis induced by dimethylnitrosamine and promoted by Clonorchis sinensis infection. RESULTS: Three different "oval" cell types are identified in portal and/or periportal areas: 1) Small periductal cells with abundant heterochromatin and scant cytoplasm that are negative for AFP, CK19, OV-6 and GST-p (primitive oval cells); 2) Glycogen-rich cells, positive for AFP, but negative for CK19, OV-6 and GST-p mainly adjacent to ductal plates (hepatocyte-like oval cells); and 3) small cells with desmosomes and basement membrane, containing GST-p CK19 and OV-6 but negative for AFP, present in ducts (ductular-like oval cells). It appears that C. sinensis infection stimulates proliferation and differentiation of small ductular or periductal cells (primitive oval cells) into either hepatocyte-like oval cells, which mature into hepatocytes without malignant transformation, or into ductular-like oval cells. CONCLUSIONS: We propose that the ductular-like oval cells are precursors of dysplastic ductular cells that give rise to cholangiocarcinomas after dimethylnitrosamine treatment and conclude that primitive oval cells are bipolar progenitor cells for hepatocytes and biliary cells, and that activation (initiation) of these cells by carcinogen (dimethylnitrosamine), followed by stimulation of proliferation of biliary cells by C. sinensis, promotes primitive oval cells or their progeny (ductular-like oval cells) to transform into cholangiocarcinomas.

Metagonimus miyatai sp. nov. (Digenea: Heterophyidae), a new intestinal trematode transmitted by freshwater fishes in Japan and Korea.

Metagonimus miyatai n. sp. (Digenea: Heterophyidae) is described based on adult flukes collected from experimental dogs and hamsters fed with metacercariae encysted in the sweetfish, Plecoglossus altivelis, dace, tribolodon hakonensis and T. taczanowskii, common fat-minnow, Morocco steindachneri, pale chub, Zacco platypus, and dark chub, Zacco temmincki, and on those collected from naturally infected humans. The new species was morphologically compared with M. yokogawai and M. takahashii obtained from experimental animals fed with the sweetfish and the crucian carp, Carassius carassius, respectively. The uterine loops of M. miyatai reached near the posterior end of the body through the space between the two testes, whereas those of M. yokogawai, occupied only the space between the acetabulum and anterior border of two testes. This uterine tubule distribution was similar to that of another closely related species, M. takahashii. However, vitellaria of M. miyatai ended in front of the posterior end of the left testis, while those of M. takahashii reached the posterior end of the left testis and ran it over. By raising M. miyatai as a new species, differentiation of M. yokogawai and M. takahashii became very clear. A key to the species of the genus Metagonimus in the Far East has been proposed.

Conger myriaster, a new second intermediate host of Heterophyopsis continua (Digenea:Heterophyidae).

Six metacercariae were found from the gill filaments of Conger myriaster purchased at Mokpo-shi in Korea on 7 September, 1996. Based on the morphology of the excysted specimen, we identified them as metacercariae of Heterophyopsis continua. C. myriaster is a new intermediate host of H. continua in the literature.

The infestation states and changing patterns of human infecting metacercariae in freshwater fish in Kyongsang-do and Kyonggi-do, Korea.

The infestation rates and abundances of human infecting metacercariae (Clonorchis sinensis, Metagonismus spp., Centrocestus armatus, Echinostoma hortense, Echinochasmus japonicus, Clinostomum complanatum) in freshwater fish were investigated at the three river areas-Taewhagang (river), Hyongsangang (river), Nakdonggang (river) -in Kyongsang-do and at four streams-Yonpungchon, Munsanchon, Kyonganchon, and Konjiamchon-in Kyonggi-do, Korea in 1994-1995. The fish caught at Taewhagang were heavily infested with metacercariae of Clonorchis sinensis and Centrocestus armatus. At Hyongsangang, Zacco platypus and Z. temmincki were moderately infested with metacercariae of C. armatus. Chomanpo, at the basin of Nakdonggang, was still endemic for C. sinensis. In the fish caught at four streams of Kyonggi-do, metacercariae of C. sinensis exhibited the highest infestation rate and intensity out of 6 species of metacercariae. The infestation intensity of C. sinensis metacercariae in fish flesh was markedly different according to each division of flesh. The cause of this difference was conjectured as a result of larval behavior. The metacercariae of C. armatus were found in almost all parts, except scales and fins, of fish. The infestation rates and intensities of C. sinensis and C. armatus metacercariae in Taewhagang greatly increased as compared with those of previous reports. Rhinogobius brunneus and Acanthorhodeus macropterus are newly recorded intermediate hosts of Echinostoma hortense. The reason of large differences from previous data was discussed and the standard method of metacercaria examination was proposed.

Classification and host specificity of Metagonimus spp. from Korean freshwater fish.

Taxonomic problems of Metagonimus spp. in Korea were investigated. Metacercariae of various freshwater fish species -- Plecoglossus altivelis, Carassius auratus, Zacco platypus, Zacco temmincki, Opsariichthys bidens -- were collected from different localities in Korea and experimentally fed to golden hamsters. Observation of recovered adult worms showed that Plecoglossus altivelis was infected with metacercariae of both M. yokogawai and M. takahashii. C. auratus was infected with metacercariae of M. takahashii and Z. platypus, Z. temmincki, O. bidens were infected only with metacercariae of Metagonimus Miyata type. From the inferences about the morphological characteristics, host specificities and occurrence patterns in infected animals, Metagonimus Miyata type is considered to be an independent group.

Gorgoderid trematodes (Digenea: Gorgoderidae) from the urinary bladder of frogs in Korea.

Two species of the family Gorgoderidae (Trematoda: Digenea), Gorgodera japonica Yamaguti, 1936 and Gorgoderina bombinae Yu & Lee, 1983 were collected from the urinary bladder of frogs captured from various localities in Korea. The morphology of each species is described and illustrated. Gorgodera japonica differs from G. cygnoides by having deeply branched vitellaria. Gorgoderina bombinae is considered a valid species.

Promoting role of Clonorchis sinensis infection on induction of cholangiocarcinoma during two-step carcinogenesis.

Chronic Clonorchis sinensis (CS) infection is etiologically related to cholangiocarcinoma (CHCA) in human and animals. This study was carried out to clarify the role of CS infection on dimethylnitrosamine (DMN)-induced cholangiocarcinogenesis. Fifteen hamsters were administered with 15 ppm DMN for 4 weeks and one week later, the hamsters were infected with 15 metacercariae of CS (DMN-->CS group). The other 15 hamsters were infected with CS and after 5 weeks they were treated with the drug, praziquantel. Again one week later, the hamsters were administered with DMN (CS-->DMN group). The other 15 hamsters were administered with DMN and CS simultaneously (CS+DMN group). Histopathological examination of the livers showed CHCA with papillary or adenomatous hyperplasia of bile ductules in 3 of 15 hamsters of DMN-->CS group and in 11 of 15 hamsters of DMN+CS group. These results suggest that CS infection to hamsters may have a promoting effect on the development of CHCA.

Morphologic descriptions of Taenia asiatica sp. n.

Among taeniid tapeworms infecting humans through pork or beef, Taenia solium Linnaeus 1758 and Taenia saginata Goeze 1782 have already been known. Based on the morphologic characteristics of adult and metacestodes of Asian Taenia saginata, the third kind of human taeniid tapeworm known to distribute in Asian countries, a new species name of Taenia asiatica is proposed. In addition to the known biology in their intermediate hosts, T. asiatica was different morphologically from Taenia saginata Goeze 1782 in having the unarmed rostellum on the scolex of adult, the large number of 'uterine twigs' and the existence of 'posterior protuberance'. These structures in the gravid proglottids were used as taxonomic keys in taeniid tapeworms for the first time. T. asiatica metacestode (Cysticercus viscerotropica) was different morphologically from T. saginata metacestode (Cysticercus bovis) in having wartlike formations on the external surface of the bladder wall.

Effect of Clonorchis sinensis infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian golden hamsters.

The study was carried out to observe the effects of Clonorchis sinensis infection on induction of cholangiocarcinoma in Syrian golden hamsters to which 15 ppm dimethylnitrosamine (DMN) solution was administered for 8 weeks. The histopathological changes of the bile duct and liver cells were observed at the 11th week. In six of 8 hamsters (75%) which were treated with DMN and then infected with C. sinensis, the livers developed cholangiocarcinoma at 10 weeks after the infestation of C. sinensis. The features of cholangiocarcinoma lesions were adenomatous or papillary hyperplasia of the bile duct epithelia showing distinct anaplastic changes with mucinous cell metaplasia and necrotic area. In the hamsters which received either DMN or C. sinensis alone, the livers showed only hyperplastic changes of the bile duct epithelial cells. It was suggested that C. sinensis infection and DMN administration could be a synergism on the development of cholangiocarcinoma in Syrian golden hamsters.

Trematodes of the genus Haematoloechus (Digenea: Plagiorchiidae) from frogs in Korea.

Total 242 Rana nigromaculata and 9 R. catesbeiana were collected from the various localities in Korea from February 1989 to July 1991, and their lungs were examined. Five species in genus Haematoloechus, i.e., H. sibiricus japonicus (Yamaguti, 1936), H. nanchangensis Hsiung, 1934, H. variegatus (Rudolphi, 1819), H. lobatus (Seno, 1907), H. lobatus koreanus, were identified in this study. Among them, H. lobatus koreanus was a new subspecies and H. variegatus and H. lobatus were newly recorded from Korea.