As a series of systematic classification for Korean common liver fluke, Fasciola sp., karyotype was investigated by means of the modified air-drying technique and of the regular Giemsa staining, Also, C-staining method was applied for detailed karyological analysis from the germ cells of the fluke. The following is a brief summary of the leading facts gained through the experiment. Korean Fasciola sp. was classified into three types based on their chromosomal complements; individuals with 20 or 30 chromosomes and with a 20/30 mosaic constitution. Worms having 30 chromosomes represent a triploid form with 3 sets of 10 basic chromosomes, while those with 20 chromosomes were diploid and mosaic individuals were 2n/3n mixoploid. The frequency of the individual type calculated is as follows; 67.45% of 212 flukes examined was of diploid, 10.85%, triploid, and the rest, 21.7%, mixoploid, respectively. In many cases, two or three types were found in the peculiar bovine host while single type inhabitant was about 20% out of 52 cases. The twenty chromosomes consisted of 1 parir of large metacentrics, 4 pairs of medium-sized subtelocentrics, and 5 pairs of small submetacentrics, while constitution of the thirty chromosomes was nearly interpreted as a triploid form with 3 sets of 10 basic chromosomes. The high centromeric indexes of both types are the first pairs among all the examined, and 37.93% was of diploid and 47.93%, triploid, respectively. In mixoploid individuals, constitution of the chromosomes of diploid or triploid cells was the same as that of diploid or triploid individuals. All the chromosomes of the germ cells in both types showed C-band around the centromeic region and especially the chromosomes no's 3, 7, and 8 showed a remarkable C-band distinguished from other chromosomes. The variance for the sizes of the worms and the eggs were not parallel with three different genotypes in Korean common liver fluke.
parasitology-helminth-trematoda ; Fasciola sp. ; karyotype ; taxonomy

Fascioliasis has not been confirmed as a human disease entity until now in Korea despite of sporadic discovery of ova of Fasciola sp. in human fecal materials being never traced to the confirmation of infection. Almost all of the cases with ova in their stool have been related with consumption of cattle liver whether eaten in raw or processed. The present authors confirmed a human fascioliasis case who was a Korean housewife of 42-year-old living in Seoul, during the exploratory laparotomy. The patient had been healthy until October 1975 when abrupt onset of urticaria, dyspepsia, epigastric discomfort developed. And the fluctuation of these symptoms was followed by epigastric colicky pain attacks from December 4, 1975. A complete worm of Fasciola sp. was removed during the bile-duct exploration with stone forceps in lower half of common bile duct, on January 20, 1976. The patient only agreed that she had eaten some raw liver of cattle on September 1975 but denied any possible sources of infection such as drinking of untreated water, handling of water flower and grass, and eating of raw watercress. The measurements of the removed worm: 35. 61 mm in body length, 14.00 mm in maximum body width(Length/width ratio, 2.54:1), distribution of testes to body length 33.9 per cent , number of branches of ovary 22, the size of intrauterine ova 157.2 x 108. 4 micrometer in average. These findings are not compatible with the classical descriptions of both Fasciola hepatica and F. gigantica, and it was concluded it is so-called Fasciola sp. which is intermediate between two species as proposed by many Japanese workers.
parasitology-helminth-trematoda ; Fasciola hepatica ; Fasciola gigantica ; Fasciola sp. ; case report ; fascioliasis ; laparotomy ; bile-duct

In an attempt to investigate the specific antigenic substance of Fasciola hepatica, Ouchterlony tests and immunoelectrophoretic analyses were carried out. Crude Fasciola antigen was prepared and fractionated by Sephadex G-200 column to Antigen I, II and III according to protein content. Crude antigens of Paragonimus westermani, Clonorchis sinensis and Paramphistomum sp. were also prepared for control and absorption study. Antiserum was prepared by injecting 0.5 ml of crude Fasciola antigen with same amount of complete Freund's adjuvant in rabbits, 10 times at an interval of l week. The results obtained in this study were as follows: Crude Fasciola antigen reacted with antiserum with 9 precipitin bands by Ouchterlony test and with 11 bands by immunoelectrophoresis. Cross reaction was observed between Paragonimus, Clonorchis and Paramphistomum antigens and anti-Fasciola rabbit serum respectively. By Ouchterlony test, 3-4 cross reacting bands were found. Anti-Fasciola sera which were absorbed with respective Paragonimus, Clonorchis and Paramphistomum antigens, reacted with Fasciola crude antigen. Ouchterlony test gave 5-6 precipitin bands. Further reaction between Fasciola antigen and antiserum absorbed with the above 3 antigens concomitantly gave 5 precipitin bands by Ouchterlony test and 7 bands by immunoelectrophoretic analyses. Fractionated Fasciola antigens (Antigens I, II and III) reacted with anti-Fasciola rabbit serum in immunoelectrophoresis. Antigen I, II and III gave 2, 3 and 5 precipitin bands respectively. Anti-Fasciola rabbit serum which was absorbed with 3 trematodes antigens gave, by immunoelectrophoresis, 1 band with Antigen I, 2 bands with Antigen II and III of Fasciola hepatica. From the above results, it is concluded that Fasciola hepatica possessed the specific antigenic substance not cross-reacted with other trematodes.
parasitology-helminth-trematoda ; Fasciola hepatica ; Paragonimus westermani ; Clonorchis sinensis ; Paramphistomum sp. ; antigen ; immunology ; electrophoresis

The authors would record the fifth case of human fascioliasis in Korea, occurring in a 48 year old Korean man who lived in Munkyong, Kyongsangbuk-do. The diagnosis was based on the morphology and measurements of the eggs which were collected in necrotic debris from the tunnel-like ulcerations of the gallbladder wall which was surgically removed for a gall stone. Although we failed to find out the fluke in the specimen, there found an evidence of once presence of Fasciola in the inflamed wall of the gallbladder in this patient.
parasitology-helminth-trematoda ; Fasciola sp. ; fascioliasis ; Heterophyes dispar ; case report ; praziquantel ; gallbladder

A case of ectopic fascioliasis involving cecum and ascending colon was reported. This case was a 19-year old Korean female who had a history of postprandial aggravation of intermittent abdominal pain of 3 weeks duration. She was presented with an abdominal mass in the right upper quadrant, for which an operation(right hemicolectomy) was performed. Whole cecum and ascending colon showed multifocal tunnel-like necrotizing granulmas with central cavitations. Numerous Charcot-Leyden crystals were seen along tracts of the worm. The lesion was distributed throughout the entire thickness of the wall. Parasitological examination confirmed the worm found in the intestinal wall to be juvenile form of Fasciola species. Possibilities were discussed on the localization of the fluke and the mode of infection of present case.
parasitology-helminth-trematoda ; Fasciola sp. ; fascioliasis ; case report ; cecum ; colon ; granuloma

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciola hepatica, peroxidase of conjugate anti-cattle IgG and orthophenylenediamine as a substrate by micro-method technique of Voller et al. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows: In assay for optimal dilution of stock antigen, the antigen (protein contents; 0.8 mg/ml) was diluted from 1/50 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y=-0.181-0.00127X in infected sera, and log Y= -0.578- 0.000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. In assay for optimal dilution of sera, the sera were diluted from 1/25 to 1/400 with in PBS/ Tween 20(pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y=-0.1540-0.0007238X in infected sera and log Y=-0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). In the 27 cases of negative intradermal test, OD values of the ELISA are 0.447 +/- 0.144, the 95 percent confidence interval (Mean+2 x SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100 percent in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was 0.846+0.224(-0.224). The 75 percent (44 cattle) among them had higher value with compared to the criterion, and the 60 percent (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. Prevalence of fascioliasis was 43.4 percent in the application of the ELISA to 272 cattle which were reared in Jeonbug district.
parasitology-helminth-trematoda ; enzyme-linked immunosorbent assay ; Fasciola sp. ; fascioliasis ; cattle

To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10-15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7 kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10 kDa bands. Two bands in sparganum extract (130 and 64 kDa) and two bands in hydatid cyst fluid (52 and 27 kDa) were cross-reacting bands with sera from cysticercosis patient. Saline extracts of Fasciola, Clonorchis and Paragonimus did not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7 kDa.
parasitolgy-helminth-cestoda ; Taenia solium ; cysticercus ; brain ; enzyme-linked immunoelectrotransfer blot ; Fasciola sp. ; Clonorchis sinensis ; Paragonimus westermani

In an attempt to investigate the antigen-antibody relations and the value of immunodiagnosis for several helminths, Ouchterlony tests and immunoelectrophoreses were carried out. Taenia saginata, Cysticercus sp. of cestodes, Clonorchis sinensis, Fasciola hepatica and Paramphistomum sp. of trematodes,and Ascaris suum of nematodes were used as antigens. On the other hand, antisera were obtained by injecting 0.5 ml each of the above antigens and the same amount of complete Freund's adjuvant into rabbits ten times at an interval of one week. The result obtained in this study are as follows: A larger number of precipitin arcs were demonstrated in homologous antigen-antibody reactions than in heterologous antigen-antibody reactions both in Ouchterlony tests and immunoelectrophoreses. Gross reactions were observed between the different species of the same class, but no cross reactions were noticed when the classes were different with one or two exceptions, such as between T. saginata, F. hepatica and A. suum. In A. suum, the difference between the male and female was more distinct in Ouchterlony test and immunoelectrophoresis than in the examination of organs such as genital organ and coeliac fluid. Immunoelectrophoresis revealed specific arcs and higher sensitive reaction than Ouchterlony test, and was considered to be a more valuable method for identifing species and immunological diagnosis.
parasitology-helminth-trematoda ; cestoda ; nematoda ; immunoelectrophoresis ; Taenia saginata ; Cysticercus ; Clonorchis sinensis ; Fasciola hepatica ; Paramphistomum sp. ; Ascaris suum ; antigen ; immunology

In order to obtain some informations on the nature and relative activity of the phosphatases present in various helminths, biochemical studies have been made in thirteen kinds of worm parasites including the adults and larvae (Fasciola hepatica, Eurytrema pancreaticum, Paramphistomum sp., Taenia solium, Taenia pisiformis, Dipylidium caninum, Diphyllobothrium mansoni, Cysticercus cellulosae, Cysticercus fasciolaris and Sparganum). A comparison based on the analysis of pH-activity curves was made among these helminths. The worm materials were mostly obtained alive from an abattoir and removed from the organs or tissues of the animal hosts naturally infected. Sparganum and Cysticercus cellulosae, however, are collected from the subcutaneous tissue of the patients by surgical removal. The worms thoroughly washed were weighed and transferred with 0.1 M Tris buffer to a chilled glass grinder (Capacity; 15 ml) and homogenized in the cold. The homogenate was centrifuged at 5000 RPM for 30 minutes. The supernatant was pipetted off for determination of the phosphatase activity. Incubation mixtures consisted of 1 ml substrate, 1 ml buffer and 0.5ml extract. The buffers used were Tris (Hydroxymethyl) aminomethane and citric acid monohydrate and the substrate was paranitrophenyl phosphate (1 gm/25 ml). These mixtures were incubated at the temperature of 37 C for 30 minutes in water bath. The absorbance or transferance of mixture was determined colorimetrically by "Spectronic 20 "spectrophotometer at 410 nm against a distilled water blank. The amount of phenol liberated was then calculated from a standard curve using phenol solutions. Controls consisted of unincubated mixtures. The results were deducted from this experiment. The phosphatase activity occurred over all parasitic helminths used in this experiment. In trematodes, pH-activity curves have demonstrated two peaks of phosphatase activity in Fasciola hepatica and Paramphistomum species. However the acid phosphatase activity was predominantly found and the alkaline phosphatase activity was found distinctly to be low in all three species. In Eurytrema pancreaticum, the pH-activity curves displayed two peaks in acid phosphatase activity, one at pH 5.0 and the other pH 9.0. In cestodes, both alkaline and acid phosphatase activity displayed the pH optima 5.0 and 9.0 to 10.0 in the adult tapeworms. However, major activity in the adults is due to the alkaline phosphtases. In contrast to the adults, Cysticercus and sparganum showed the higher activity in acid phosphatases which predominates in the larvae. In all cases of nematodes, the pH optimum for acid phosphatase was 4.0 to 6.0. A preponderance of acid phosphatase activity was shown in the extract of intestine of Ascaris lumbricoides. The aspect that phosphatases are correlated with phosphorylated passage of substances through the cuticle of helminths and may also be involved in carbohydrate metabolism is discussed.
parasitology-helminth-trematoda-cestoda ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum sp. ; Taenia solium ; Taenia pisiformis ; Dipylidium caninum ; Diphyllobothrium mansoni ; Cysticercus cellulosae ; Cysticercus fasciolaris ; sparganum ; alkaline phosphatase ; acid phosphatase ; biochemistry

: In order to know the species and frequency of human parasitic infection diagnosed by biopsy, 149 cases (0.18%) of parasitic infection were reviwed, which were selected from 80,947 biopsied materials submitted for routine histopathological examination during a period of 10 years from 1980 to 1989 at Department of Pathology, Chonnam National University Hospital. They consisted of 112 cases of cysticercosis, 17 paragonimiasis, 7 clonorchiasis, 4 amebiasis, 1 sparganosis, 1 enterbiasis, 1 aniskiasis, and 1 fascioliasis respectively. Based on morphological preservation of cysticercus, they could be divided into mild (20.2%), morderate (40.4%), and severe (39.4%) degeneration. Except 2 cases biopsied at the lungs, 15 cases of ectopic paragonimiasis were located at abdominal cavity (8 cases) and central nervous system (7 cases). One case of intrahepatic fascioliasis was observed. This is the 13th human fascioliasis reported in Korea. From the above results, the frequency of parasitic infections found in biopsied specimens was on the decrease as the cysticercosis and ectopic paragonimiasis.
parasitology-helminth-cestoda-nematoda-trematoda ; biopsy ; histopathology ; tissue parasite ; cysticercose ; ectopic ; Enterobius vermicularis ; Taenia solium ; Paragonimus westermani ; Clonorchis sinensis ; Entamoeba histolytica ; Anisakis ; Fasciola sp. ; paragonimiasis ; fascioliasis