For the two-dimensional observation on the growth and development of Clonorchis sinensis, an image analysis system (IBAS 2000, Kontron, Germany) was used in this study. On 3, 6, 9, 12, 15, 21, 30 and 90 days of infection, 474 worms were collected from rats infected with 50 metacercariae each. The overall recovery rate was 31.6%. The worms grew rapidly in their length and width up to 30 days of infection and then, did slowly to 90 days of infection. However, the growth pattern of body area was not similar to that of body length and width, because the body area increased continuously even after 30 days of infection. With the image analysis system, the sizes of irregular-shaped genital organs were measured easily, which showed sigmoidal growth patterns. The growth rate of genital organs increased rapidly until 21 days of infection when the uterus was filled with numerous eggs, and then gradually slowed down to 90 days. There was no difference in growth pattern between the anterior and posterior testis. The seminal receptacle, however, showed an abrupt increase in size between 15 to 21 days of infection when it was filled with condensed semen.
Therefore, the growth pattern of seminal receptacle might be used as another criteria to estimate the extent of sexual maturation of C. sinensis. From this study, it is suggested that image analysis system is very useful to reveal the growth and development pattern of C. sinensis, especially of their internal organs.
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The surface ultrastructure of metacercariae and adults of Gymnophalloides seoi, the only known gymnophallid infecting humans, was observed by scanning electron microscopy.
Metacercariae were ovoid or pyriform in shape and slightly concave ventrally. The oral sucker had two sizes of type I papillae, small and large, encircling its lip. Type I papillae were arranged in a row on both side of the body.
The ventral pit had several type I papillae on its inner surface. The ventral sucker was covered with cobble-stone like cytoplasmic processes and had 6 type I papillae on its lip. The surface of the body was covered with the tegumental spines except for the ventromedian area between the two suckers. The spines at anterior body were digitated into 3-5 points, and their size decreased at posterior one third of the body. Adult worms were rhomboid or ellipsoid in shape and covered with tegumental spines except for the ventromedian area. The shape and distribution of the tegumental spines and sensory papillae were similar to those of metacercariae. However, sensory papillae arranged in a row on the ventral surface of metacercariae were not observed in adults. The ventral pit became larger and more prominent as the fluke grew. It is suggested that the ventral pit function as an additional adhesive organ to the host tissue.
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To evaluate the feasibility of irradiation as a control measure for metagonimiasis, the metacercariae of Metagonimus yokogawai were irradiated with gamma ray, either after isolation from the sweetfish (Plecoglossus altivelis) or in situ of the fish, and their survival and development in rats were observed at 7 days post-infection. The radiation dose varied from 5 to 100 Gy for the metacercaria-irradiation group and from 5 to 500 Gy for fish-irradiation group. The results showed that the worm recovery rate from the irradiation groups decreased as the radiation dose was increased. Higher doses of radiation were required for the fish-irradiation group to obtain the same results as the metacercaria-irradiation group. The LD50 of the metacercaria-irradiation group was 4.5 Gy, whereas that of the fish- irradiation group 6.2 Gy. A few number of worms which survived until 7 days in rats were severely retarded especially in the growth of their reproductive organs, i.e., complete or partial failure in the development of testes and formation of uterine eggs. The present study revealed that irradiation of sweetfish by 200 Gy is effective to control infectivity as well as development of M. yokogawai metacercaria in rats.
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Metagonimus yokogawai was found deeply invaded into the submucosa of the small intestine of mice (ICR) when they were immunosuppressed by prednisolone injection.
Experimental groups consisted of control, fluke infection (1,800 metacercariae per mouse), and fluke infection plus immunosuppression. In fluke infection group, many worms were found sectioned in the intervillous space of the jejunum and ileum at 6 hrs, 12 hrs, and 1 day after infection, and pathological changes characterized by villous atrophy and crypt hyperplasia were observed. After 3 days, only a few worms were found in intestinal sections, and after 7 days, the pathological changes became minimal. No worm was found penetrated beyond the mucosal layer. On the other hand, in immunosuppressed mice, numerous worms were found sectioned in the duodenum and jejunum, irrespective of the infection period up to 14 days. Pathological changes of the mucosa were minimal until 3 days after infection, but at 5 days marked destruction of the mucosal layer was observed. At this time many flukes were found invaded deeply into the submucosa facing the muscular layer. Despite continuous immunosuppression, the mucosal damage was gradually recovered at 7-21 days post-infection. The results showed that immunosuppression of ICR mice can induce, for a short period of time, severe mucosal damage, and allow deep invasion of M. yokogawai into the submucosa of the small intestine.
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Protease activity was identified in crude extracts of Paragonimus westermani eggs which were purified from infected dog lungs, isolated on 14 weeks after metacercarial challenge. The eggs were used after removing possibly contaminated host or worm tissues on their shell surfaces.
In the crude egg extracts, high proteolytic activities against carboxybenzoyl-phenylalanyl-arginyl-4-methoxy-beta- naphthylamide (Cbz-phe-arg-MNA) and Azocoll were detected whereas those against succinyl-alanyl-prolyl- phenylalanyl-p-nitroanilide (Suc-ala- pro-phe-pNA) were not revealed. The enzyme exhibited the maximal activity at pH 6.
Its activity was inhibited by specific cysteine protease inhibitors, 10(-5) M 1-trans-epoxysuccinylleucylamido (4- guanidino) butane (E-64) and 1 mM iodoacetamide (IAA) while potentiated by 6.5-fold in the presence of 2.5 mM dithiothreitol (DTT). When the enzyme was purified partially by Sephacryl S-300 High Resolution gel filtration, it migrated as a single homogeneous band at 35 kDa. The 35 kDa cysteine protease has been recognized neither in the metacercariae nor in the adult. These findings indicated the presence of at least one protease of cathepsin family in immature eggs of P. westermani.
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Activities of different cysteine proteases of Paragonimus westermani in cleaving human IgG Y B Chung, H J Yang, S Y Kang, Y Kong, S Y Cho The Korean Journal of Parasitology.1997; 35(2): 139. CrossRef
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Interstrain polymorphisms of isoenzyme profiles and mitochondrial (Mt) DNA fingerprints were observed among seven strains of Acanthamoeba isolated from different sources and morphologically assigned to A. polyphaga. Mt DNA fingerprints by eight restriction endonucleases (Bgl II, Sca I, Cla I, EcoR I, Xba I, Kpn I, Sal I, and Sst I) revealed considerable interstrain polymorphisms. Isoenzyme profiles revealed considerable interstrain polymorphisms for acid phosphatase, lactate dehydrogenase, and glucose-6-phosphate dehydrogenase while those for glucose phosphate isomerase, leucine aminopeptidase, and malate dehydrogenase showed similarity. Despite of the interstrain polymorphisms, the isoenzyme profiles and Mt DNA fingerprints of the strain Ap were found to be identical with those of the strain Jones.
Mt DNA fingerprinting was found to be highly applicable for the strain identification, characterization, and differentiation.
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Bacterial endosymbiosis within the cytoplasm of Acanthamoeba lugdunensis isolated from a contact lens storage case D I Chung, H H Kong, T H Kim, M Y Hwang, H S Yu, H C Yun, S Y Seol The Korean Journal of Parasitology.1997; 35(2): 127. CrossRef
Close relatedness of Acanthamoeba pustulosa with Acanthamoeba palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns Young Ho KIM, Mee Sun OCK, Ho Cheol YUN, Mee Yul HWANG, Hak Sun YU, Hyun Hee KONG, Dong Il CHUNG The Korean Journal of Parasitology.1996; 34(4): 259. CrossRef
PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga H H Kong, D I Chung The Korean Journal of Parasitology.1996; 34(2): 127. CrossRef
Biochemical and molecular characterization of a strain KA/S2 of Acanthamoeba castellanii isolated from Korean soil D I Chung, H H Kong, H S Yu, Y M Oh, S T Yee, Y J Lim The Korean Journal of Parasitology.1996; 34(1): 79. CrossRef
Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish Kyung-il Im, Ho-Joon Shin The Korean Journal of Parasitology.2003; 41(4): 181. CrossRef
We have investigated the action of sodium nitrite on the growth and morphologic changes of T. vaginalis and on the treatment of subcutaneous abscess by trichomonad in mice.
Sodium nitrite inhibited the growth of metronidazole-sensitive KT9 isolate and metronidazole- resistant CDC85 strain of T. vaginalis as concentration of 6 mM and 10 mM, respectively. Intraperitoneal injection of sodium nitrite (70 micrograms, 100 micrograms, 130 micrograms/g body weight) did not reduce the size of abscess produced by subcutaneous inoculation of T. vaginalis in mice. T. vaginalis, treated with sodium nitrite at concentration giving about 50% inhibition of growth, showed fissures, many vacuoles and electron-translucent zone in the cytoplasm by transmission electron microscopy. In the case of CDC85 treated with 9 mM sodium nitrite, hydrogenosomal matrical change, destruction of hydrogenosomal membrane, autophagic vacuoles, disappearance of Golgi complex and polysome were notably observed. With above results, it is assumed that sodium nitrite inhibits the growth of metronidazole- sensitive and--resistant strains of T.
vaginalis and induces the morphological changes of T.
vaginalis although it does not affect in reducing of abscess size by T. vaginalis in mice.
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The protozoan parasite, Entamoeba histolytica, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to E. histolytica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the finding that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to E. histolytica trophozoites.
Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to E. histolytica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polymerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 x 10(7) molecules/micrograms of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA). Lysates of E. histolytica also induced expression of mRNA for IL-8 in colon epithelial cells. These results suggest that acute inflammatory reaction by E. histolytica may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.
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Antigenic localization in Paragonimus iloktsuenensis worm tissues (tegument, intestine and vitelline gland) in different developmental stages of 2 weeks, 3 weeks, 4 weeks, 5 weeks and 33 weeks from albino rats (Sprague-Dawley) infected with P. iloktsuenensis was observed by electron microscopy. These worm tissues of different developmental stage of P. iloktsuenensis was observed on electromicrograph by immunogold labeling method using P. iloktsuenensis infected rat serum of 10 weeks. Antigenic localization was demonstrated as labeling of gold particles in tissues on electronmicrograph. In tegument, gold particles were labeled on tegumental tissue, generally more numerous on secretory granules in tegumental syncytium 2 weeks than those on the other elder developmental stages, but there was a little variation in antigenicity according to individual worm tissue. In general, antigenicity in tegumental tissue was not strong (gold particles: 0.1- 5/1 microns 2). In intestine, a large number of gold particles (15-18/1 microns 2) were labeled in intestinal epithelium. Gold particles were concentrated especially on secretory granules in cytoplasm, and gold particles were labeled not only in cytoplasmic protrusions, but also in intestinal luminal contents. Intensity of labeling of gold particles was not correlated with developmental stage of worms. In vitelline gland, a large number of gold particles were labeled on vitelline globules. The gold particles in vitelline globules (8-11/1 microns 2) were concentrated in protoplasm among segmental globules.
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Immunoelectron microscopic localization of partially purified antigens in adult Paragonimus iloktsuenensis Ok-Ran Lee, Pyung-Rim Chung The Korean Journal of Parasitology.2001; 39(2): 119. CrossRef
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Three-week-old ICR SPF mice were orally inoculated with one of 5 doses ranging from 2 x 10(2) to 2 x 10(6) oocysts of Cryptosporidium muris (strain MCR) per mouse. Oocyst inoculation was directly proportional to the amount of oocysts shed and was inversely proportional to the period required for peak oocyst production and to the prepatent period. Peak oocyst production occurred between fifteen and thirty-one days with a patent period from 61 to 64 days.
Three days after all mice stopped shedding oocysts, they were orally challenged with a single dose of 2 x 10(6) oocysts of the same species. Marked seroconversion for IgG antibody accompanied recovery from mice inoculated with 5 x 10(5) oocysts. Mice administered with carrageenan excreted a small number of oocysts for 49.0 days on the average after challenge inoculation (ACI) and control mice for 14.2 days in a dose-independent fashion. Just before challenge infection, phagocytic activity of peritoneal macrophages (M phi) and the number of peripheral M phi were dramatically decreased. Mild challenge infection implies that the immunogenicity of C. muris (strain MCR) is very strong, despite M phi blocker carrageenan administration.
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Schistosomiasis is a snail-transmitted trematodiasis acquired by immersion in water which contains the cercariae.
In Korea, six imported cases of urinary schistosomiasis by Schistosoma haematobium and one case of imported cerebral schistosomiasis by S. mansoni were reported. Herein we report a case of S. mansoni infecting rectum of a 46 year-old Korean male, who had been to Saudi Arabia for two years. On colonoscopy for routine physical check up, a 0.4 cm polyp in the rectum was detected and biopsy was done.
Microscopically, rectal mucosa showed several granulomas which were composed of macrophages, lymphocytes, neutrophils and eosinophils. The center of each granuloma showed an ovoid egg often containing miracidium. The eggs measured 130 x 60 microns in average size. They had yellowish-brown transparent shell with the characteristic lateral spine.
This is the 8th imported case of schistosomiasis in Korea and the second one infected by S. mansoni.
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In the present communication, YS-27, a Korean strain of Entamoeba histolytica is described for the isolation and establishment of axenic cultivation. E. histolytica, designated as strain "YS-27", was isolated from the pus of a hepatic abscess obtained from a 72 year old inpatient of August 10, 1969. Specimens, were obtained by needle aspiration, inoculated immediately and weekly cultured in a modified diphasic medium at 37 degrees C. Strain YS-27 had been maintained for more than 15 years by weekly subculture until February, 1985. These cultures were transferred to a monoxenic TTY-SB medium seeded with a trypanosomatid of the genus Crithidia. Penicillin G, 2 to 10 x 10(3) International units and Streptomycin, 2 to 10 mg per 100 ml, were added to the cultures to eliminate the bacteria. After more than one year later, these two organisms were well maintained by transfer every 3 or 4 days until January, 1986 at 37 degrees C in TTY-SB medium in the absence of other microorganisms.
These monoxenic cultures were then transferred to TYI-S-33 medium. Strain YS-27 alone had not been growing at the time of transfer, but when overlaid with Crithidia at intervals of 3 to 4 days, strain YS-27 propagated well. The Crithidia died out several weeks later after several passages.
Beginning in April, 1986, strain YS-27, was successfully established in axenic culture in TYI-S-33 medium and has been maintained in continuous culture and multiplied well to present.
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