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Detection of circulating antigens in rats experimentally infected with Paragonimus westermani by ELISA (ELISA법을 이용한 폐흡충 감염 백서의 혈청내 항원 검출)

  • 용태순;김동수
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.141-148
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    • 1987
  • Circulating antigens in rats experimentally infected with Paragonimus westermani were examined by ELISA. From a total of 22 albino rats, each fed with 25 metacercariae, blood samples were collected until 12 weeks after infection. The specific antibodies against P. westermani in the serum of an infected cat were purified by ammonium sulfate precipitation, DEAE anion-exchange chromatography and affinity chromatography serially. So-called double antibody sandwich ELISA method was used for the detection of circulating antigens. The results were as follows: Mean value of O.D. in control sera was O. 04 (S.D.=0. 04). After infection, mean O.D.(S.D.) values were changed serially: 0.03(0.01) at 0.5 week(3 days), 0.55(0.50) at 1 week, 0.69(0.45) at 1.5 week, O.20 (0.19) at 2 weeks and O.13(0.10) at 2.5 weeks of infection. They returned, thereafter, to the level before infection. When O. 16 (mean+3 S.D.) were considered as cut-off value, those higher than O. 16 were observed only in the sera collected between 1 and 2.5 weeks after infection. Average 8. 4 immature worms (2.2 from the lungs and pleural cavities; 6.2 from muscles) were recovered in a rat at 12 weeks after infection. The fact that circulating antigens were not detected after 3 weeks of infection was considered to the caused by the formation of antigen-antibady complexs.

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Isolation and identification of Cyuptosporidium from various animals in Korea (한국산 동물로부터 크립토스포리디움의 분리 및 동정 III. 닭으로부터 Cryptosporidium baileyi의 분리)

  • Lee, Jae-Gu;Seo, Yeong-Seok;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.315-324
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    • 1991
  • Each of SPF chicken(Hi-Line strain, 2-day-old males) was inoculated with 2.5 or $5\times10^4$ oocysts by stomach tube. The oocyst was the medium type of Cryptosporidium previously isolated from Korean chicken origin, and passed in 2-day-old SPF chicken. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of the developmental stages, the bursa of Fabricius of the chicken was examined by transmission electron microscopy (TEM) on the 12th day postinoculation. The prepatent period for 8 chicken was 5.9 days postinoculation on the average, and the patent period was 12.9 days. The number of oocysts discharged per day for the chicken was reached peak on day 12 postinoculation on the average. A large number of oocysts was found in fecal samples obtained from inoculated chicken on days 8~14 postinoculation. The ultrastructural feature of almost every developmental stage of the medium type from chicken was very similar to that of Cryptosporidium previously isolated from mammalia including human and birds except for the attachment site of C. tsuris to the mucus cell from mammalia, but dimension of the oocysts from fecal samples of the medium type was different from those of C. meleagridis and mammalia origin. The above results reveal that the medium type of Cryptosporidium of Korean chicken origin is identified as Cryptosporidium baileyi.

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Epidemio-entomological survey of Japanese encephalitis in Korea (한국에 있어서 일본뇌염의 역학적, 매개동물학적 조사)

  • 백두현;주종윤
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.67-86
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    • 1991
  • In order to determine the seasonal prevalence and population dynamics of Culex tritaeniorhynchus in relation to the epidemics of Japanese encephalitis, and ecology of these vector mosquito in Kyungpook Province, Korea, studies were con- ducted during the Period of 7 years from 1984 to 1990. Cx. tritaeniorhynchus first collected in June between 4th and 28th, and trapped in large numbers during the period from mid-August to early September, showed a simple sharply pointed one-peaked curve. There was a gradual decrease from mid-September, with a very small number of them collected until early October in every year. The average number of Cx. tritaeniorhynchus rapidly decreased after 1985, and the number became particularly low in 1989. The highest population density, which was observed in August during the initial three years, was found to be delayed in the following years, accompanied by a decrease in the number of mosquitoes. In the trend of nocturnal activity of Cx. tritaeniorhynchus, with oncoming darkness they become very active, gradually decreasing in activity toward mid night, but slightly increasing toward dawn. The immature stages of Cx. tritaeniorhynchus were first found in rice fields contributing to peak adult densities in mid-July. The highest average densities of Cx. trisaeniorhynchus was 14,900 per m2 on mid-August 19th. The larval Cx. tritaeniorhynchus showed high resistance levels and resistance ratios against 5 organophosphorus compounds. In the adult horisontal life table characteristics of Kyungsan colonies of Cx. tritaeniorhynchus under insectary condi- tions, life expectancy was 28.3 days for males and 59.8 days for females. The net reproductive rate was 7.8 and generation time was 25.6 days.

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Natural killer cell activity in Naegzeria fowleri infected mice (Naegleria fowleri 자연살세포의 활성에 미치는 영향)

  • Lee, Gyu-Rae;Sin, Ju-Ok;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.267-278
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    • 1991
  • The natural killer (NK) cell activity of splenocytes and recycling capacity of NK cells were observed by combining the $^{51}Cr-release$ cytotoxicity assay and single cell cytotoxicity assay against YAC-1. The ICR mice were infected intranasally with Naegleria fewleri, that is a pathogenic free-living amoeba. The mice infected with $1{\times}10^5$ trophosoites showed mortality rate of 76.7% and mean survival time of 12. 9 days. The cytotoxic activity of NK cells in infected mice was significantly higher than that of non-infected mice during the period between 12 hours and day 3 after infection, and highest on day 1. The target-binding capacity of NK cells in infected mice was not different from that of non-infected ones. Maximal killing potential and maximal recycling capacity were remarkably increased in infected mice as compared with the control. The results obtained in this observation indicated that elevated NK cell activity in mice infected with N. fowieri was not due to target-binding capacity of NK cells but due to the increased activity of NK cells and increased recycling capacity of individual NK cells.

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Epidemiological Survey on Metugonimus yokogawui Infection in the Eastern Coast area of Kangwon-Province, Korea (강원도 동해시 황강흡충 감염의 역학적 조사연구)

  • 안영겸;정평림
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.59-68
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    • 1987
  • An epidemiological survey on Metagonimus yokogawai infection was performed in the eastern coast area of Kangwon province. Formalin-ether concentration technique and Stoll's dilution egg counting method were employed to figure out the prevalence and intensity of M. yokogawai infection and eating habits of inhabitants with raw freshwater fish. The results are summarized as follows: 1. The prevalence rate of M. yokogawai infection was 6.6% among a total of 2, 357 examinees; 8.6% (115 out of 1, 345) in male and 4.0% (40 out of 1, 012) in female. 2. The prevalence rates were different by the streams where the examinees were residing: 17.8% along the area of Ohsip-cheon (stream), 11.8% along Maeup-cheon, 1.7% along Hwasang-cheon, 1.3% along Yeongok-cheon, 0.9% along Namdae-cheon, 0.9% along Sa-cheon and 0% in the other streams (Yongchon-cheon, Kangnung Namdae-cheon and Jusoo.cheon) in decreasing order. 3. By social strata, the positive rate was 8.0% (122 out of 1, 521) in general inhabitants and 3.9% (33 out of 836) in school children. 4. By the intensity of infection by means of EPG counts in feces; the light infection (less than 400/EPG) was 74.1%, moderate infection (401-1, 000/EPG) 17.3% and heavy infection (more than 1, 001/EPG) 8.6%. 5. Twelves out of 50 sweetfish (Plecoglossus altivelis) (24%) caught from the streams in eastern coast of Kangwon-Do were infected with metacercariae of M. yokogawai, and the number of larvae detected were 70.7 per 6sh in average. The metacercarial infection rates in fish were also different by the streams; 100% in Ohsip-cheon of Samcheok-Gun, 14.3% in Sa-cheon, 16.7% in Yeongokcheon of Myeongju-Gun and 0% in Namdae-cheon of Yangyang-Gun, respectively. 6. Six hundred ninety live (695) out of 1, 396 inhabitants and school children (49.8%) were experienced in eating raw freshwater fishes (Plecoglossus altivezis, Triboledon hakonensis, Coreoperca sp., Morose sp., Carassius sp. & Cyprinus sp. and Zacco platypus). The data were assayed with the questionnaire collected from the examinees. In summarizing the above results, it is known that the Ohsip-cheon area of Samcheok-Gun is highly infected region, and the other areas of Kangnung Namdae-cheon, Yeongok-cheon and Yangyang Namdae-cheon are very lowly infected or non-infected regions for M yokogawai infection.

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Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis (간흡충 충체의 부위별 항원성에 대한 면역조직화학적 연구)

  • Jin Kim;Jong-Yil Chai;Weon-Gyu Kho;Kyu-Hyuk Cho;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.21-30
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    • 1991
  • An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, rising formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000∼1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1 : 2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyme portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.

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Effect of Aflatoxin $B_1$ on Ultrastructural Changes of Biliary Epithelial Cells in Mice Experimentally Infected with Clonorchis sinensis (간흡충 감염 마우스에 있어서 환관 상피세포의 미세구조에 대한 Aflatoxin $B_1$의 영향)

  • 민홍기
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.99-109
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    • 1987
  • The present study was carried out to examine the effect of a calcinogen, aflatoxin B1 on the ultrastructural changes of ciliary epithelial cells in mice infected with Clonerchis sinensis. A total of 93 male albino mice(BALB/c strain) was divided into 3 groups; group I, treated with 1.0 ppm aflatoxin Bl for 12 weeks; group II, given 50 C. sinensis n;etacercariae, and group III, given 50 metacercariae and treated with 1.0 ppm aflatoxin Bl for 12 weeks. Three mice served for untreated-uninfected controls. From 4 weeks after the treatsment and/or in(ection, three mice from each group were sacrificed at 4 week intervals up to the 40th week, and their hepatobiliary tissues were prepared for transmission electron microscopy. The most prominent ultrastructural changes in group I were remarkable enlargement of nuclear size, separation of nucleolus, dispersed chromatin granules in nuclei and increased dense granules along the inner membrane of nuclei. In the cytoplasm there was slight proliferation of mitochondria and endoplasmic reticulum (ER) at earlier stage. At the 12th week separation of fibrillar and granular components of the nucleolus was a characteristic finding. As the time elapsed, epithelial cells showed fiattened-cuboidal form and a tendency of atrophy. Most of the nuclei were elongated and polygonal in shape. In group II the appearance of elaborate interwoven folds of lateral cytoplasm forming a labyrinth of interconnected intercellular space and variety in nuclear shape were the prominent fadings at earlier stage. The cytoplasm showed slight proliferation and dilatation of mitochondria and ER, and a small number of mucin droplets. In the basement membrane scanty fibrous cells were seen. With time, variety in nuclear shape, marked proliferation and dilatation of rough ER and some collagen fibrils were demonstrated. Other features of intracellular organelles and mucin droplets persisted. In group III cuboidal epithelial cells showed their remarkably enlarged and irregular nuclei, increased chromatin granules in the nuclei, separated nucleoli, proliferated and dilated rough ER. With time, sequestered mitochondria showed blob-like evaginations which lacked cristae and dense matrix, and were limited by a single membrane. Since the 20th week, microvilli were relatively scanty and poorly developed. Organelles and inclusions in the cytoplasm of metaplastic cells were poor. Nuclei were variable in shape. The nlost prominent changes at later stage were separation of nuclei from the cytoplasm, and appearance of numerous and irregularly angled electron dense granules in the nuclei.

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Blastogenic responses of splenic Iymphocytes to Naegleria fowleri Iysates and T-cell mitogen in mice with primary amoebie meningoencephalitis. (실험적 뇌막수염에 있어 Naegleria fowleri 항원에 대한 세포매개성 면역 반응)

  • Park, Gwang-Min;Ryu, Jae-Suk;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.1-6
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    • 1987
  • This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.

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Passive Transfer of Immunity against Clonoychis sinensis by Peritoneal Exudate Cells in Mice (복강삼출세포를 이입받은 마우스에서의 간흡충에 대한 면증응답)

  • Gwon, Tae-Chan;Gang, Jin-Mu;Choe, Dong-Ik
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.45-50
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    • 1987
  • This study was undertaken to evaluate the role of peritoneal exudate cells in the transfer of immunity against the liver cuke, Clonorchis sinensis in the inbred BALB/c mice. Ten donor mice were divided into 2 groups. One group consisted of 5 mice was infected orally with 20 metacercariae of C. siitensis, and the other group was injected intraperitoneally with 20 excysted larvae. Thirty days after immunization, the peritoneal exudate cells tore obtained from the donor mice. Twenty recipient mice were divided into 4 equal groups for the purpose of primary immunization. The mice of Group I were injected intraperitoneally with $2{\times}10^6$ peritoneal exudate cells of the donor mice infected orally, those of Group II were injected intraperitoneally with $2{\times}10^6$ peritoneal exudate cells of the donor mice injected intraperitoneally. Those of Group III were injected orally with 20 metacercariae of C. sinensis. The group IV mice served as controls. Four days after the primary iMmunization all recipient mice were challenged orally with 20 metacercariae of C. sinensis, and then killed 30 days after the challenging infection. When the peritoneal exudate cells were injected into the recipient mice, pronounced reduction in eggs per gram of the feces was found in the mice o( Group I and Group II, but no reduction in those of Group III. In the worm burdens of C. sinensis, the number of flukes found in the mice of Group II was only significantly less than those in the control group (IV). In addition the number of plaque forming cells per spleen in the mice of Group II was found larger than those in Group I. It is likely that donor peritoneal exudate cells transferred to the recipients might result in the production of relative immunity.

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Intestinal flukes of genus Metagonimus and their second intermediate hosts in Kangwon-do (강원도에 분포하는 Metagonimus속 장흡충과 제2중간숙주에 대하여)

  • 안영겸
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.331-340
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    • 1993
  • This study investigate the epidemiological feature of Metagonimus infection in Kangwon-do (province). The average Infection rate of the surveyed inhabitants was 7.8% (83 positives out of 1, 067 examinees) by stool examination; male, 11.4% and female, 3.2%, respectively. The egg positive rate in residents in the Som river area was 7.3%, that of the Chuchon river area 6.3%, the Pyongchang river area 12.8%, the Tong river area 3.8%, the Hongchon river area 9.8%, and the Ohsip stream area of Samchok 8.0%,respectively. The average metacercarial infection rate of genus Metagonimus in the fish was 81.0% (256 positives out of 318 fish). The infected fleshes were Zacco platypus. Zacco teminki, Opswiichthys biens, Squdidis sp., Corqssius carassius, etc. in western Kangwon-do Meanwhile, in the Ohsip stream area of Samchok-gun, eastern costal Kangwon-do, the infected fish were Plecoglossus altivelis and Tribolodon hokonensis. The rats and dogs are infected with the metcercanae obtained from Zacco platypus and Opsariichthys biens, adult worms collected were Miyata type of Metagonimus with some M. takahashii. When infected with metacercariae from Plecoglossus ltivelis, Metagonimus yokogowai was only found. M. yoogawai and Metagonimus Mlyata type were fecund together in Tribolodon hakonenis in Ohsip stream area of Samchok, in the eastern Kangwon-do. The intestinal flukes of genus Metogonimus in western Kangwon-do were Miyata type of MetQnonimuT and M. takahashii transmitting mainly by Zacco platypus and Opsariichtys bidens as a source of infection. In the eastern part of Kangwon province (Ohsip stream area of Samchok), M. yokogowai was mainly distributed by P. altivelis and T. hakonesis, but some T. hakonensis harbored the metacercariae of Miyata type of Metagonimus with those of M. yokogawai.

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