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

In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis (SDS-PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblotted. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable.
parasitology-helminth-trematoda ; Paragonimus westermani ; immunology ; antigen ; electrophoresis

The effect of praziquantel on P. westermani exposed in vitro was observed by stereomicroscope, light microscope and scanning electron microscope. Following results were found. The worms incubated in 0.01 micro-g/ml praziquantel were moving after 26-hour incubation. However, all of them were immobilized immediately after incubation in solutions over 0.1 micro-g/ml concentration. All of the exposed worms showed severe vacuolization not only in tegument but in subtegument, intestine, ovary, testis, Mehlis' gland and excretory bladder. Vacuoles in tegument burst out to form craters. As incubation time went on, tegumental structure was disintegrated severely. The worms exposed to praziquantel were observed to be immobilized and be vacuolized of all tissues. Disintegration of reproductive organs suggests that praziquantel have suppressive effect on egg production when the flukes are not killed. The drug effects were found more related with incubation time than with drug concentration.
parasitology-helminth-trematoda ; Paragonimus westermani ; electronmicroscopy ; chemotherapy ; praziquantel

Total of 2,643 Korean soldiers were examined of their stool for parasitic infections by both cellophane thick smear and formalin-ether concentration techniques from August 1983 to December 1985. Out of them, 73.6% were free from any parasite, 22.6% were ova positive and 4.0% cyst positive. The ova positive rates by species were Ascaris lumbricoides 2.0%, Trichuris trichiura 13.0%, hookworm and Trichostrongylus orientalis 0.08% respectively, Clonorchis sinensis 7.6%, Metagonimus yokogawai 1.1%, Paragonimus westermani 0.08%, Echinostoma hortense 0.04%, Fibricola seoulensis 0.9%, Taenia sp. 0.3%, Hymenolepis nana 0.2% and H. diminuta 0.04%. Most of them (87.3%) were positive by a species, 11.9% by two and 0.8% by 3 species. The cyst positive rates were Entamoeba histolytica 0.6%, E. coli 1.4%, Endolimax nana 1.7%, Iodoameba butschlii 0.04% and Giardia lambla 0.9%. Among the cyst positives, 89.5% were positive by a species, 7.6% by two and 0.5% by 3 species. The intestinal parasite infections among the Korean soldiers decreased distinctly compared with previous data.
parasitology-helminth-nematoda ; trematoda ; cestoda ; protozoa ; epidemiology ; Clonorchis sinensis ; Trichuris trichiura ; Metagonimus yokogawai ; hookworm ; Ascaris lumbricoides ; Diphyllobothrium latum ; Hymenolepis nana ; Entamoeba histolytica ; Giardia lamblia ; Entamoeba coli ; Endolimax nana ; Trichostrongylus orientalis ; Paragonimus westermani ; Echinostoma hortense ; Fibricola seoulensis ; Taenia sp. ; Hymenolepis diminuta

A study on the population density of crayfish intermediate hosts and infestation status of crayfish with encysted larvae of Paragonimus westermani in Ulchin county, Kyungpook province, Korea was conducted from May to October in 1986. The population density of the crayfish ranged from 1 to 13, with an average of 4 per manhour. Among the six habitats, Ducheon had a somewhat higher density than that of the others. Of eight hundred and seventeen crayfish examined, 127 or 15.5 per cent harboured the metacercarial larvae of Paragonimus westermani. The majority of the larvae were found in three parts of the body: most frequently in the cephalothorax, next in the gills, and then in the liver. The average number of metacercarial larvae per infected crayfish ranged from 1.0 to 1.9, with an average of 1.7. Summarizing the results, this study indicates that the population density of crayfish intermediate host and infestation rates for the crayfish with encysted larvae of Paragonimus westermani in Ulchin county of Kyungpook province is relatively high.
parasitology-helminth-trematoda ; Paragonimus westermani ; metacercaria ; epidemiology ; cray fish-Cambaroides similis

Enzyme-linked immunosorbent assay (ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for l hour at 10,000 rpm at 4C. Affinity-purified antigen (antibody-bound antigen) was prepared from fractions (bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum (6 months infected) obtained by ammonium sulfate (40-45 per cent saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34,700,27,600 and 11,500 in molecular weights. All cats were divided into five groups(G1-G5) by different worm burdens. The mean of recovered worms (+/-SD) and the number of cats in each group are as follows:G1, 2 worms(0) and 4 cats; G2, 4.75(+/-0.66) and eight; G3, 10.75(+/-1.92) and four; G4, 25.20(+/-3.43) and five; G5, 48(+/-12.63) and five cats. The results were summarized as follows: The antibody levels(OD value) increased by worm burden in G1 to G4 generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in G4 and G5. These results suggested that the worm burden in G4 (about 20 - 30 worms) is enough to produce antibody maximum in cats of 2~3 kg weight. The antibody levels increased significantly (p<0.05) compared to control sera at the 3rd week in G1 and G2, at the 2nd week in G3, and at the 1st week in G4 and G5. Especially in the 4th week, OD value increased more in G1(p<0.001) and in G2 to G5(p<0.01). In the pattern of antibody levels by ELISA in each group, OD in G1 increased to the 18th week continuously, in G2 OD was maintained same after the 16th week, but in G3 it decresed after the 16th week, and it was maintained same in G4 and G5 after the 14th week. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In G1 and G2 OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in G3 than that with crude antigen to the 16th week and OD of G4 and G5 were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity-purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen.
parasitology-helminth-trematoda ; immunology ; enzyme-linked immunosorbent assay ; Paragonimus westermani ; paragonimiasis ; cat

This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100 mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of 2 microgram/ml antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at 51-65 per cent precipitate (PA2), followed by 0-50 per cent precipitate (PA1), 66-80 per cent precipitate (PA3), and 81-90 precipitate (PA4). Unprecipitated protein over 90 per cent ammonium sulfate (PA5) showed the lowest antigenicity. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. By passing through the Sephadex G-200 column, PAl and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2- I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. SDS-polyacrylamide gel electrophoresis of PA1-I, PA1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons (KD) to 196 KD; of them 220 KD protein was major band. Fraction PA2- I was composed of 255-225 KD, and PA3-I, 255-240 KD, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. Absorbance by ELISA began to increase within 10-20 days after the infection and reached the highest on 140-180 days, then made plateau thereafter. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I , fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at 20-30 days after infection.
parasitology-helminth-trematoda ; immunology ; enzyme-linked immunosorbent assay ; Paragonimus westermani ; antigen

To evaluate the immature eggs of Paragonimus westermani as a source of diagnostic antigen, about a million eggs which were excreted by 104 adult worms were collected; their saline extract (soluble egg antigen; PwSEA) was prepared. The specific IgG and IgM antibody levels were observed in experimental dog paragonimiasis by micro-ELISA, using PwSEA as well as whole worm extract of 12 week-old P. westermani (PwWWE). The protein composition of the PwSEA was observed by disc-PAGE. The results could be summarized as follows: Specific IgG antibody to PwSEA began to increase on 8 weeks after the experimental infection; it maintained its high level until the observation period of 13 weeks. The levels of IgM antibody to PwSEA, however, did not show any significant change. Specific IgG antibody to PwWWE began to increase earlier from 2 weeks after the infection and continued to increase until the observation period of 13 weeks. Its level was much higher than that to PwSEA. Specific IgM antibody to PwWWE increased temporarily during 2-8 weeks after the infection. By disc-PAGE, PwSEA showed 2 protein bands of very low motility. The bands of PwSEA corresponded to the first and second bands in the electrophoretic pattern of PwWWE of the 12 week-old worms. The above results indicated that the PwSEA induced antibody production in dog paragonimiasis but its antigenicity was weaker than PwWWE to be used as a diagnostic antigen.
parasitology-helminth-trematoda ; immunology ; Paragonimus westermani ; antigen ; enzyme-linked-immunosorbent assay ; serum ; IgG

In order to observe the protein compositions of soluble extracts of P. westermani, and their changes during early developmental stages, the crude saline extracts of 4, 6, 8, 10 and 12 week-old worms which were harvested from experimentally infected dogs were analysed by disc-PAGE. The results were as follows: A total of 15 bands were identified from electrophoregrams of respective developmental stages. Of them, 5 bands were recognized throughout the developmental stages. The number and protein amount of identified bands changed according to the worm development from 4 weeks to 12 weeks. However, the banding patterns of 4 and 6 week-old worms and 8 and 10 week-old worms were similar each other. Of 15 identified bands, band 1 was recognized only in 12 week-old worms whereas bands 3, 4, 8, 9, 10, 11 and 15 gradually lowered their amount according to development to disappear in 12 week-old. In addition, band 5 became a major band in 12 week-old while band 6 turned to a minor band at tha same age. The possible relations of changing patterns of protein bands with worm development were discussed.
parasitology-helminth-trematoda ; Paragonimus westermani ; immunology ; protein ; electrophoresis

The present study was performed to observe the ultrastructure of the integument of adult Paragonimus westermani. Dogs experimentally infected with 60 metacercariae of P. westermani were autopsied 4 months after the infection. Adult P. westermani were extracted from the dogs and the fine structure was studied by means of scanning and transmission electron microscope. The findings are as follows: Scanning electron microscopic findings showed that the spines and the papillae are distributed at whole body surface but the well developed spines or papaillae are shown around the oral suckrer and ventral sucker. At the end of the body, excretory pore was found, the shape was irregular. Transmission electron microscopic findings showed that plasma membrane, tegument, basal lamina, connective tissue, circular muscle layer, longitudinal muscle layer, nerve axon and tegumental cell were observed. In higher magnification, plasma membrane and bar-shaped granules were found at the outer surface of the tegument.
parasitology-helminth-trematoda ; Paragonimus westermani ; electron microscopy