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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.109-118
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    • 1989
  • Production of circulating specific antibodies to the lung fluke (Paragenimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods, However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of p. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs(male and female), and eggs. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um thickness as the antigen and cat antisera (11~20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobensidine(DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1 : 500~1 : 2, 000 and 1 : 200~1 : 500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

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Cross-reacting and specific antigenic components in cystic fluid from metacestodes of Echinococcus grannlosus and Taenia solium (포충 및 유구낭미충 낭액에 있어서 공통항원 및 특리이원 분획)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho;Duk-Young Min
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.131-140
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    • 1989
  • Sera from confirmed patients of 5 hydatidosis, 67 neurocysticercosis and 89 other parasitic diseases were tested for specific antibody (IgG) levels by ELISA to cystic quid antigens from metacestodes of Echinococcus granuzosus (HF) and Taenia sodium (CF). All hydatidosis sera reacted positively to both HF and CF while neuro- cysticercosis sera did in 49.3% to HF and 85.1% to CF, The frequencies of cross- reactions were lower in other parasitic diseases to both antigens, By SDS-PAGE, protein bands of 64, 35, 22 and 7 kilodaltons (kDa) were found common in HF and CF. SDS-PAGE/immunoblot exhibited that hl·datidosis sera reacted crossly to CF at 135, 110, 100, 86, 64, 45, 39, 35 and 24 kDa bands while neurocysticercosis sera did to HF at 135, 100, 86, 64, 52, 39, 35, 29 and 24 kDa bands. These results indicated that protein bands of 135, 100, 86, 64, 39, 35 and 24 kDa were major common components in HF and CF. Protein bands of 7 kDa in HF and 15, 10 and 7 kDa in CF did not react crossly and were specific components in respective antigens.

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An experimental study on prednisolone-induced interstitial pneumonia caused by Pneumocystis carinii (프레드니솔론 투여에 의한 조폐포자충(Pneumocystis carinii)성 간질성 폐염에 대한 실험적 연구)

  • 신대환;이영하;나영은
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.101-108
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    • 1989
  • This study was performed to observe the role of Pneumocystis carinii as an etiologic agent of interstitial pneumonia in immunocompromised hosts. Total 90 male Sprague-Dawley rats, approxi. mately 150-180 g, were used. Fifteen of them were used as control group and remaining 75 (5 groups) were as immunosuppression groups; group 1 received prednisolone (25 mg/kg twice weekly) only; group 2 Prednisolone and tetracycline (75 mk/kg/day) ; group 3 Prednisolone, tetracycline and trimethoprim-sulfamethoxasole (50~250 mg/kg/day) : group 4 prednisolone and trimethoprim-sulfamethoxasole; and group 5 prednisolone and griseofulvin (300 mg/kg/day) until death. The survival days of each group rat were calculated, and upon death their lungs were removed immediately and then stamp smears were prepared and stained by Giemsa or toluidine blue O. For histopathologic observation, lungs were fixed in 10% formalin, cut into sections and stained with Gomori's methenamine silvei, hematoxylin-rosin, and Brovkn & Brenn stain. The results obtained were as follows: 1. The mean survival time of each group rat was 19.3$\pm$5.2 days (group 1), 41.1$\pm$14.0 days (group 2), 50.5$\pm$18.4 days (group 3), 43.0$\pm$22.9 days (group 4) or 21.8$\pm$5.1 days (group 5). Significant differences were noted between group 1 and group 2(p<0.01), group 1 and group 3 (p<0.01), and group 1 and group 4 (p<0.01), which represented bacterial infections were most fatal in immunocompromised rats. Group 5 revealed no difference in the survival day from group 1, while significant differences were noted between group 2 and group 5(P<0.01), group 3 and group 5(p<0.01), and group 4 and group 5(p<0, 01), which represented little importance of fungal infection as the cause of death of the rats. 2. The first fatality due to p. carinii pneumonia occurred 17 days after the beginning of the immunosuppression. The occurrence rate of P. carinii pneumonia in the decreasing order was 92.9% (group 3), 80.0% (group 2 and group 5), 78.6% (group 4) and 33.3% (group 1). With regard to the pathological stage of P. carinii pneumonia, the stage 1 was 11.3%, the stage 2, 28.3%, and the stage 3, 60.4%. 3. Viewing from the duration of immunosuppression, bacterial pneumonia chieay appeared in 1 month, mixed infections (P. carinii and bacteria, or p. carinii and fungi) in 1~2 months, and pure P. carinii pneumonia after 2 months. The present study revealed that P. carinii pneumonia was the most important cause of death of immunocompromised rats later than 1 month after the start of immunosuppression.

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Differentiation of Entomoeba histolyticn and Entcmoeba dispor in cyst-passers by immunoblot (면역이적법을 이용한 아질아메바와 동형아메바의 감별진단)

  • 이미정;홍성태
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.247-254
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    • 1996
  • Differentiation of invasive strains of Entamoebn histolytica according to their pathogenicity has been a topic of long debate, but now the pathogenic species only is regarded as E. histolytica while the non-pathogenic species is E. dispar. The present study applied immunoblot to differentiale infections of the two species among microscopically- detected cyst-passers in Korea. The crude extract of 5. histolyticn separated in 5-20% gradient gels, revealed many fractions of 94. 81. 71, 50. 44, 38.5. 37.5, 29, 19. and 18 kDa when the cysteine proteinase inhibitor. E64, was supplemented. The serum IgG antibody of 3 proven E. histolytirc cases reacted loth the antigenic fractions of 117. 110. 99.68,66,60.54.52, 46. and 45 kDa. Sera of PCR confirmed 3 cases of E. disper reacted only to the 117 kDa fraction or the E. histolytica crude extract which was regarded as non specific. To the antitigen of monoxenic E. dispar. sera or E. dispar and E. histolytica cases showed the same immunoblot reactions. The serum IgG antibody reacted with several antigenic fractions of both E. histolytica and E. dispar. but IgM and IgE antibodies showed no reaction to either antigen. Sera of 24 symptomless amebic cyst-passers were screened with the E. histolytica alltigen; two were found to be infected by E. histolytica and 22 were by E. dispar. The present findings suggest that in Korea most of asymptomatic cyst passers of E. histolytica are carriers of E. dispar. Immunoblot using E. histolytica antigen is a good technique for the differentiation of E. histolytica and E. dispar infections.

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Effect of Splenectomy on Development of Primary Amoebic Meningoencephalitis (비장적출(splenectomy)이 원발성 아메바성 뇌수막염의 발생에 미치는 영향)

  • Sin, Ho-Jun;Im, Gyeong-Il;Choe, Rim-Sun
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.156-164
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    • 1985
  • To elucidate the effect of splenectomy on the development of experimental primary amoebic meningoencephalitis in mice, the death rate and survival time of mice infected intranasally with Naegleria fowleri trophozoites $5{\times}10^4$ cultivated in CGVS medium were compared according to the mouse age when splenectomy was done, and post-operation until experimental infection. Immunodigusion was undergone to detect the presence of serum antibod). due to N, fowleri infecttion in mice. Polyacrylamide gel electrophoresis was done to compare the protein fractions of mouse serum in each experimental groups. In experiment I, splenectomy was done 3 weeks and infection 4 weeks after birth, the death rate of control, sham operated and splenectomized group were 100%, 85% and 95%, and the mean survival time after infection 7.3 days, 7.5 days and 7.8 days, respectively. In experiment II, splenectomy was undergone 3 weeks and infection 6 weeks after birth, the death rate of of control, sham operated and splenectomized group were 95%, 95% and 95%, and the mean survival time after infection 12.1 days, 11.5 days and 11.5 days, respectively. In experiment III, splenectomy was done 5 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95%, 90% and 95%, and the mean survival time after infection 8.1 days, 8.3 days and 8.5 days, respectively. By Ouchterlony immunodigusion, anti-JV. fowleri antibody in the serum of mouse with primary amoebic meningoencephalitis was detected against a N. fowleri antigen, which was prepared by ultrasonication of N, fowleri trophozoites, each reacting two lines of precipitation. The patterns of serum fractions by polyacrylamide gel electrophoresis were different between control and sham operated groups from splenectomized group in fraction II, III and V, the sera of which were collected after N. fowleri infection. This results may be summarized as that splenectomy has no effect on the development of primary amoebic meningoencephalitis in mice.

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Studies on the Intestinal Fluke, Metagonimus yokogawai Batsurada, 1912 in Korea IV. Geographical Distribution of Sweetfish and Infection Status with Metagonimus Metacercaria in South-Eastern Area of Korea (한국의 요꼬가와흡충(Metagonimys yokogawai)에 관한 연구 IV. 동남부 지방산 은어의 지리적 분포 및 요꼬가와흡충 피낭유충의 감염상태)

  • Song, Cheol-Yong;Lee, Sun-Hyeong;Jeon, Sang-Rin
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.123-139
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    • 1985
  • The present survey was undertaken to observe the geographical distibution of matacercaria among the sweetfish, Plecoglossus altivelis, the most important second intermediate host of Metagonimus yokegawai, which were collected during the period from 1983 to 1984 in south-eastern coastal areas of Korea. The results obtained were summarized as follow: 1. A total of 668 sweetfish collected from 70 chins (=streams) in the above coastal arcus was examined for the metacercariae, and 432 (64.7%) among them were infected with metacercariae of M. yokogawai. The mean number of metacercariae in a fish was in the range from 0 to 29,604 and the mean number of the metacercariae in a fish was 640.3. 2. It was observed that the mean infection rate showed 46.45 in Gangweon-Do (=province) . The infection rates varied from stream to stress showing 100% in Namdae stream(Gangnun-city) , Obsib stream (Samchok-gun) and Dongmak, 95.0% in Hosan stream, 90.0% in Gungchin stream, and Namdae stream (Yangyang-fp) ,80.0% in Hwasang stream,50.0% in Daebong stream,45.0% in Yonghwa stream, 30.0% in sangchsn stream and Sachsn stream, 20.0% in Munam stream and Okgyoe stream, 15.0% in Imweon stream, 10.0% in Namchsn stream. And no metacercarial infections in their fish host were found in the areas of Mulchi stream, Sangun stream, Gosamun stream, Jonchon stream and Gagok stream. 3. It was observed that the mean infection rate showed 50.9% in Kyangsangbuk-Do (=province). The infection rates also varied from stream to stream showing 100% in Songchon stream, Osib stream (Yongdok-gun) and Daejong stream,60.0% in Pyanghae stream and Gunmu stream,50.0% in Buhwung stream and stream, 25.0% in Nagok stream, 20.0% in Oangbi stream, 10.0% in Namhodong stream. And no metacercarial infections in their fish host were found in the areas of Namdae stream (Uljinup) and Chaksan stream. 4. It was observed that the mean infection rate showed 67.6% in Kyangsangnam-Do(=province). The infection rates varied from stream to stream showing 100% in Taehwa river, Hwiya river, Inchfn stream, Miryang river, Nam river, Hwagae stream, Koha stream, Sosang stream, and Tongchon stream,90.9% in Yancho stream,90.0% in Hoihwa stream,80.0% in Chindong stream, 66. YoA in Chingyo stream, 40.0% in Hoyam river. And no metacercarial infections in their fish host were found in the areas of Chinjon stream, Sanchan stream and Jukch6n stream. 5. It was observed that the mean infection rate showed 94.6"A in 0h511anam-Do(=province). The infection rates varied from stream to stream showing 100% in DongchEn stream, Isachon stream, Somjin river(GoksEng-gun) , Ssmjin river(Kurye-gun) , S6si stream, Gokchsn stream, Kohung stream, Kwansan stream, Youi stream, stream, Unj6n stream and Apnae stream, 92.3% in Tamjin river, 90.9% in Okkok stream, 84.6% in Songgen stream. And no metacercarial infections in their fish host were found in the area of Ypmgpl stream. 6. On the other hand, the infection rate showed 100% in Inchsn river of Chollabuk Do (=province) (=province) and Kangsn stream of Cheju-Do(=province) .

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Seasonal Variations of Metacercarial DEnsity of Clonorchis sinensis in Fish Intermediate Host, Pseudorasbora parva (간흡충 유행지역에서 참붕어내 피낭유충 감염밀도의 계절적 변동)

  • Gang, Sin-Yeong;Kim, Seok-Il;Jo, Seung-Yeol
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.87-94
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    • 1985
  • The seasonal variations of the rate and intensity of metacercarial infection of C. sinensis in P. parva were observed. The fish were collected at Sun-Am River which located in Kim-Hae City, Kyong-Sang Nam Do (=Province), Korea, from March 1983, to February 1984 every month. A total of 788 fish was examined. The number of metacercariae in each fish was individually counted after the individual digestion by artificial gastric juice. The results were as follows: 1. During one year, 513 (65.1%) out of 788 fish were infected with metacercariae. In May, June, July and September, the infection rates ranged from 82.0% to 98.6% whereas the rates was relatively low in March, April, November and February ranging from 11.4% to 64.7%. 2. The intensity of infection was similar with those of infection rates. The mean intensity per infected fish was 103.0 and standard deviation was 118.9 throughout one year. The highest mean intensity was in June (294.8) and the lowest in November (11.1). 3. The observed frequency of fish with certain intensities of metacercariae were fitted to theoretical equations derived from negative binomial distribution in March, April, November and February (p>0.05). Meanwhile, the equation of lognormal distribution were fitted with the observed frequencies in May, June, July and September (p>0.05, p>O. 75). The variance/mean ratio varied by month. The value was the highest in July (814.3) and the lowest in November (158.8). Unlike our hypothesis, the metacercarial density of Clonorchis sinensis in its the most favorable fish host, Pseudorasbora parva showed considerable seasonal variations in the hyperendemic area. The possible factors were discussed.

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Effect of anthelmintics on the early stage of Enterobius vermicularis (요충 초기 발육단계에 대한 구충제의 효과)

  • 조승열;강신영
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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    • pp.7-17
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    • 1985
  • In order to determine the susceptible age of Enterobius vermicular is to anthelmintics and to observe the chronologie growth of female E. vermicularis in man, experimental infections were done. About 500 eggs were challenged to 19 volunteers. After 4, 8, 16, 20, 24, 28, 32 and 35 days of infection, each case was treated by either mebendazole or pyrantel pamoate. On the 40th day of infection all cases including control were treated again to terminate the experimental infection and to evaluate the effect of previous treatment. Each case collected 3-day stools to harvest the expelled worms. The results could be summarized as follows: 1. The infection rates of females were in range of 0.6~13.1 % in control cases. Because the collected worms showed comparable growth and development by day, the worms were concluded to be derived from experimental infection. 2. Cases that were treated with mebendazole on 4, 8 and 16 days after infection expelled 37.5%, 2.5% and 67.5% of the number expelled by a control case on the 40th day. Cases treated thereafter expelled no worms on the 40th day. 3. Cases that were treated with pyrantcl pamoates on 4, 8, 16, 24, 28, 32 and 35 days, expelled 90.7%, 25%, 45.3%, 8%, 2.7%, 5% and 29.3% of the number collected from control cases in respect. 4. All the worms collected were females. The total body length increased consistently and comparably from the 20th day of infection. Those collected on the 20th day were 2.5~3.0 mm long with vagina, sac-like structure and strands of ovaries; 24 day-old worms may have short uterus, 28 day-old worms had long uterus without eggs, 32 day-old worms began to produce eggs, 35 day-old worms showed wide variations in egg deposit in uterus, and 40 day-old worms had uterus filled with eggs from vulva to anal levels. From the above results, it was inferred that the life span of female Enterobius vermicularis was longer than 40 days, and the developmental stages of worms younger than 16 days resisted considerably to both mebendazole and pyrantel pamoate.

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Effects of the Applications of Slaked Lime and Silicate Fertilizers on the Seedling Growth and Yield of Grass/Clover Mixed Swards on Newly Reclaimed Hilly Soil (신개간 산지토양에서 소석회 및 규산질 비료의 시용이 혼파목초의 초기생육 및 수량에 미치는 효과 비교)

  • Jung, Yeun-Kyu
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.23 no.1
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    • pp.23-30
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    • 2003
  • This pot experiment was conducted to find out the effects of slaked lime($L_{0}$ ; 0.00, $L_1$; 3.75, $L_2$; 7.50, $L_3$; 11.25g/pot) and silicate fertilizers($S_1$; 3.75, $S_2$; 7.50, $S_3$; 11.25g/pot) on the seedling growth and yield of grass/clover mired swards on newly reclaimed hilly soil, and it's differences between both the soil improvers. The results obtained are summarized as fellows: Comparing with the $L_{0}$ treatment without soil improver, the seedling vigour and yields of both forages were markedly enhanced by the application of soil improver. The positive offsets of soil improvers tended to be relative higher on ladino clover than orchardgrass in mixed swards. The seeding vigour, botanical composition, and yield of ladino clover were more markedly influenced by the application of silicate fertilizer than slaked line. The yields of ladino clover were increased 75.8% by the 51 over $L_1$, and 32.0% by the $S_2$ over $L_2$, respectively.

Spatial Variation in the Reproductive Effort of Mania Clam Ruditapes philippinarum during Spawning and Effects of the Protozoan Parasite Perkinsus olseni Infection on the Reproductive Effort (여름철 산란기에 있어 바지락 번식량의 공간적 변이와 기생 원생생물 Perkinsus olseni 감염이 바지락 번식에 미치는 영향)

  • Kang, Hyun-Sil;Hong, Hyun-Ki;Yang, Hyun-Sung;Park, Kyung-Il;Lee, Taek-Kyun;Kim, Young-Ok;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.37 no.1
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    • pp.49-59
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    • 2015
  • Spatial variation in the reproductive effort of Manila clam Ruditapes philippinarum is often closely associated with variation in the seawater temperature and food availability, which determines gonad maturity and the quantity of gamates produced during spawning. Previous studies also have reported that severe infection by the protozoan parasite Perkinsus olseni exerts a negative impact on clam reproduction, retarding gonad maturation or decreasing the reproductive effort. In the present study, we investigated impacts of P. olseni infection on the reproductive condition of Manila clam during a spawning season. Histology revealed that 54% of female clams in Wando off the south coast were in spawning, while only 10% of the female from Gomso and 0% of the female from Seonjaedo in Gyeonggi bay off the west coast were engaged in spawning at the end of May in 2004. Ray's fluid thioglycollate media (RFTM) assay was applied to assess P. olseni infection and indicated that the infection intensity in Wando ($3,608,000{\pm}258,000cells/g$ wet tissue) was significantly higher than the levels in Gomso ($1,305,000{\pm}106,000cells/g$ wet tissue) and Seonjaedo ($1,083,000{\pm}137,000cells/g$ wet tissue, p < 0.001). The size of the ripe female follicle determined from histology was significantly smaller in Wando ($0.032mm^2$) compared to the sizes in Gomso ($0.059mm^2$) and Seonjaedo ($0.052mm^2$, p < 0.05). Accordingly, the number of ripe eggs in the follicle was significantly fewer among clams in Wando (14) compared to the numbers determined in Gomso (23) and Seonjaedo (22). The absolute quantity of egg in ripe clams from Wando (31.01 mg) was also significantly smaller than Seonjaedo (61.79 mg) and Gomso (133.3 mg). Quantity of total protein, carbohydrate, and lipid in the tissue in the Wando samples was significantly smaller than the quantities determined in Gomso and Seonjaedo (p < 0.001). The observed poor reproductive condition and proximate tissue composition of the females in Wando were, in part, explained by the extremely high level of the parasites, sapping the ability to store energy in the host tissues, which is used in tissue growth and the egg production.