Immunoelectron microscopic localization of partially purified antigens in adult Paragonimus iloktsuenesis

An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of D1 fraction (D1A) among 5 antigenic protein fractions partially purified by DEAE-anion exchange chromatography from water-soluble crude antigen (PIWA) of adult Paragonimus iloktsuenensis. Immune reactions of adult worm tissues with rabbit serum immunoglobulin immunized with crude antigen (PI-Ig) and D1 antigen (D1-Ig), as well as rat serum immunoglobulin infected with P. iloktsuenensis were observed. D1A showed strong antigenicity in the intestinal epithelium of the worms during the early infection period of 2-4 weeks after infection. The vitellaria also showed stronger antigenicity than the other tissue sites in immune reaction of tissues against all immunoglobulins from 4 to 33 weeks after vitelline development. Therefore, it is suggested that D1A was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immuno-reactivity of two immunoglobulins (PI-Ig, D1-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with D1-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in D1 antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue.

A Study on the Effects of cognition, behavior and affection of the Music Therapy for Dementia Senile People / 성인간호학회지

The purpose of this study was to determine the effects of the music therapy, one of the dementia treatment programs, on patients' cognition, behaviors and affection, and thereby, developing a nursing intervention to help them live a quality of life. The subjects of the study were 37 dementia senial people who admitted the mental hospital at K city in chungnam province. The study was designed as repeated measures design for a single test group. The subjects were received music therapy for 40 minutes 2 times a day during 6 weeks from February 16 to March 29, 2000. The tools of study were K-MMSE developed by Park, J. H. and Kuen, Y. C.(1989), Clinical Dementia Rating developed by Morris, J(1993) at Washington university, and A fourteen symptom behavior and mood rating scale developed by Wyatt and Kupfer(1968). For the tools of music therapy, elderly favorite songs (changs, folk songs, old popular songs and children's songs) were recorded. These tapes and other tools were edited according to experts' advices. The music therapy consisted of music listening, rhythmic instrument playing and following-up singing. The therapy was performed at the lounge of the center to which the sample people could get access conveniently. A nurse was asked to observe sample people's cognition, dementia behaviors and affection, evaluated them according to the checklist scale. The collected data were processed using the SPSS WIN 7.5 program for absolute values, percentiles, mean, SD, ANOVA, GLM-repeated measure method. The items of scale and its reliability analyzed were Cronbach's alpha. The results of this study were as follows: 1. As a result of analyzing dementia elderly cognition, behaviors and affection before the music therapy, it was found that the disorder of cognition, behavior and affection was most outstanding. 2. After the therapy, the average scores of subjects' overall cognition, behavior and affection were lowered significantly. It was conceived that the music therapy should continue for 3-4 weeks. 3. Difference of cognition in test groups was significant statistically between pre-test and 2 weeks, 4 weeks, 6 weeks after test (df=3, F=6.39, p=.000), pretest and 4, 6 weeks after test (p=.01, p=.001), which suggests that a continued music therapy would be desirable to treat dementia. Difference of behaviors and affection in test groups was significant statistically between pre-test and 1-6 weeks after test (F=52.01, p = .000), pretest and 3-6 weeks after test (p=.000), which suggests that a continued music therapy would be desirable to treat dementia. 5. It was found that the music therapy was more effective in treating moderate dementia (MMSE-K score 16-23) than severe one (MMSE-K score 15 or lower), and vascular type of dementia than Alzheimer's type of one. After all, it may well be concluded that the musical therapy is an effective nursing intervention to provide the dementia elderly with a quality service and thereby, increase cognition and reduce the problematic behaviors and affection of dementia senile people.

Studies on the immunodiagnosis of rabbit clonorchiasis II. Immunoaffinity purification of whole worm antigen and characterization of egg, metacercaria and adult antigens of Clonorchis sinensis

The sensitivity and specificity of crude and affinity-purified antigens of Clonorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent assay (ELISA) The results were as follows: The antibody-binding antigen (ABA) purified from whole worm crude antigen (WWA) by CNBr-activated Sepharose 4B affinity chromatography made 4 specific bands against rabbit anti-sera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16,300-18,500 and 28,000-29,000 daltons, while major ABA protein bands were at 18,000-21,000 and 29,000-31,000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15,000-200,000 daltons, 15,000-100,000 daltons and 11,000-80,000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31,000 daltons, while those of EGA and MEA consisted of higher molecular weights than 30,000 daltons. The ELISA reactivities of WWA to rabbit anti-sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4-6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting with WWA showed a plateau without variation. MEA showed relatively low O.D. values (0.6) in heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential to increase the sensitivity and specificity for the immunodiagnosis of clonorchiasis.

ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani

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.

Comparison of agar-gel diffusion tests, counterimmunoelectrophoresis and enzyme-linked immunosorbent assay in the sera of skin test positives for paragonimiasis

Agar-gel diffusion test (AGD), counterimmunoelectrophoresis (CIEP) and enzyme-linked immunosorbent assay(ELISA) were examined with the sera of skin test positives for paragonimiasis. The crude antigen(Paragonimus whole worm extracts: protein concentration, 7.56mg/ml) and human sera were used in AGD and CIEP. And in ELISA test, diluted antigen with 1:40,000 of crude antigen and diluted sera with 1:100, 1:200 were used in the test. The positive identical ratio between AGD and CIEP reactions is 98 percent and negative identical ratio is 100 percent. One or three precipitin bands are observed in AGD. One to seven precipitin bands are also revealed in CIEP. Especially, deeply stained bands are observed in CIEP than those of AGD. The positive identical ratios between AGD and ELISA tests are 96 percent in 1:100 diluted sera, and 94 percent in 1:200 diluted sera. But the negative identical ratios between AGD and ELISA tests are 97 percent and 99 percent respectively in 1:100 and 1:200 diluted sera. The positive identical ratios between CIEP and ELISA tests are 98 percent and 96 percent respectively in 1:100 and 1:200 diluted sera, but also 97 percent and 99 percent in 1:100 and 1:200. Control sera, such as clonorchiasis, amoebiasis and toxoplasmosis, revealed all negatives with Paragonimus antigen in AGD, CIEP and ELISA tests. By above results, ELISA was most sensitive, next CIEP and AGD. But AGD test appears to be more useful when used to crude antigen without cross reaction with other parasitic infections. CIEP test is basically equal in terms of precipitin reaction, but CIEP is able to be detected more sensitively and rapidly though less simple in handiwork than AGD. Consequently, three methods for immunological tests of paragonimiasis have good correlations with one another. Also, each of these has both merits and demerits in immunological test for paragonimiasis. But the ELISA test was proved to be the most sensitive and convenient tool for mass screening test, especially in case of using purified antigen.

Changes of liver and diaphragm in experimental paragonimiasis

Changes of abdominal organs in early paragonimiasis are described in dogs and cats. Experimental infection was induced by feeding 40-50 and 20-30 metacercariae of Paragonimus westermani to 15 dogs and 15 cats respectively. These animals were killed sequentially starting from 8 days to 120 days after infection. Complete autopsies were carried out in all animals and the tissues were examined microscopically with the main emphasis on changes of the liver and the diaphragm. A definite bout of acute diffuse fibrinoexudative peritonitis with ascites ranging from 5 to 20 ml was present in both dogs and cats at the time of 20 days after infection. This was also the period of the maximum liver damage and diaphragmatic injuries. The peritonitic changes became minimized after 45 days of infection. The liver changes were of two folds. The first one was characterized by numerous pin-point or linear tissue defects on the surface of the lobes, which were often covered with fibrin clots. Some of these defects were impacted by the larvae. These 'scratch' marks appeared to be of mechanical effect, and the margins were often banal without a significant inflammatory reaction. The scratch marks were no longer observed in animals examined after 45 days. The second was noted in the Kupffer cells of the liver. A large amount of hemosiderin pigment was noted in the Kupffer cells and became prominent by 20 days through 45 days. Hemosiderin-laden macrophages were found also in the spleens of these animals. Some animals in 30 days group showed the most Kupffer cells heavily loaded with hemosiderin. However, no associated degenerative changes were noted in these cells. The diaphragm was quite unique in early phase of infection in all animals examined. Numerous pinpoint perforative lesions could definitely be seen grossly, and some of these lesions included migrating larval worms inside the tunnels. These lesions were microscopically characterized by tract or tunnel formation with accompanying tissue reaction. The diaphragmatic changes were manifest by 20 days after infection. The tracts of tunnels appeared to have been made by pressure necrosis and surrounding edema, and subsequently were associated with a massive eosinophilic influx and myocytolysis. The eosinophilic abscess and multinucleated giant cells were often observed in the animals of the 30 days group. The diaphragmatic changes seemed to be repaired with or without fibrous scar formation which was occasionally seen in 45 days and later.

Agar-gel precipitin reactions in experimental paragonimiasis

In an attempt to investigate the sensitivity of immunodiagnosis in cats experimentally infected with Paragonimus westermani, agar-gel precipitin reaction were studied. Metacercariae of P. westermani were administered to cats in various doses(2-100 metacercariae per cat) and antisera were obtained at an interval of a week. Precipitin bands appeared in homologous antigen-antibody in experimental paragonimiasis between 3 and 5 weeks after infection in all the cats. Almost all the cases in which a large number of worms were detected, showed strong reactions as revealed by deeply stained bands. Precipitin reactions did not necessarily parallel with the number of worms detected. This may be attributable to the individual difference of a cat's conditions. Very weak precipitin reactions were noticed between Clonorchis antigen and Paragonimus antisera of cats, but no reactions were noticed between Paragonimus antigen and Clonorchis antisera of cats or rabbits.

Splenic paragonimiasis: A case in a dog

A case of Paragonimus westermani cysts found deep inside the spleen is described. This spleen was from a dog that was experimentally infected via oral route with a hundred metacercariae and was killed 3 months after. At autopsy, beside the usual lesions due to Paragonimus in the lungs, the spleen was the site of multiple parasitic cysts. The involved spleen was slightly enlarged and showed an area of hemorrhagic dome on the capsular surface. There found a total of 8 adult worms in four well demarcated parasitic cysts containing a pair of worms each. All of them were found deep into the splenic parenchyme and were surrounded by a well formed connective tissue wall, thus resembling the features of the pulmonary paragonimiasis. The route of infection was assumed to be a direct transcapsular invasion during the phase of larval migration.

Analysis of protein components at varioue stages of Clonorchis sinensis

In this study the authors examined the protein components at various stages of Clonorchis sinensis, and those of tegument and metabolite of adult Clonorchis by using SDS-polyacrylamide gel electrophoresis and immunodiffusion. The following results were obtained: The protein components of C. sinensis were gradually changed during its development. A considerable change occurred during the initial 7 days after the metacercarial infection. Two bands of protein of about 97,000 molecular weight (MW) and 178,000 MW were unique to excysted metacercaria of C. sinensis. Other 2 bands of protein of 23,000 and 25,000 MW which were absent in metacercariae, might be associated with the development of sex organs in adult. In the metabolite, some components of tegumental proteins were detected. And this tegumental protein components in metabolite seems to be the major antigenic components reacting with infected rabbit antiserum by immunodiffusion. Twenty bands of protein were detected in the isolated adult tegument. Among them 6 bands were in 97,000-65,000 MW, 3 bands in 56,000-53,000 MW and 5 bands in 37,000-30,000 MW. On the other hand, in metabolic products of adult C. sinensis, 17 bands were detected.

A comparative analysis of various parts of Ascaris suum with respect to their protein composition

For the purpose of making a comparative study of protein compositions in Ascaris suum by sexes and body parts, extracts were prepared from whole bodies, body walls, genital organs, digestive organs and body fluid, of both sexes. And electrophoretic analysis was conducted using polyacrylamide slab gel in the presence of sodium dodecyl sulfate. The results were as follows: In this study, protein bands of each part were separated in the largest number and most clearly under 8 -12 percent (10 percent) gradient gel condition. The number of bands by body parts was 43 in body walls, 51 in genital organs, 47 in digestive organs, and 34 in body fluid. When examined in terms of sex, the number of bands of whole body was 38 in females and 35 in males. The electrophoretic patterns of body wall protein were in most cases similar with those seen in digestive organs. And the band with a molecular weight of 72,900 was unique to the body wall, and the 122,000 MW band was unique to the female body wall. In genital organ extracts, large molecular weight proteins (more than 80,000) were more frequently met. The molecular weights showed some differences between the two sexes. Of the bands, those having molecular weights of 119,700, 100,500, 88,500 and 86,100 were unique to the female genital organs. On the other hand, the male genital organs showed unique bands having molecular weights of 87,100, 82,800, and 15,500. An unique band common to the genital organs of both sexes was one having 49,300 MW. In the digestive organs evenly distributed protein bands of molecular weights of l0,000-120,000 were observed. The band having 59,800 MW was unique to the digestive organs. The number of bands obtained from body fluid was comparatively small, and the number of bands having less than 30,000 MW was 7, accounting for 55 percent of the total protein amounts. The bands having 47,600 MW and 31,400 MW were unique to body fluid.

Immunoelectrophoresis for Fasciola hepatica

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.

A clinical trial of oxantel and pyrantel against intestinal nematodes infections

This study was conducted to determine the effect of oxantel/pyrantel tablets, oxantel suspension, pyrantel dry syrup and pyrantel tablets against T. trichiurus and A. lumbricoides infections. Twenty-four cases of mixed infections of T. trichiurus and A. lumbricoides were treated with oxantel/pyrantel tablets at a single dose of 20 mg/kg of body weight, and 49 cases of single infection of T. trichiurus were administered oxantel suspension at a single dose of 15 mg/kg of body weight. On the other hand, single infection of A. lumbricoides in 18 cases was administered pyrantel dry syrup at a single dose of 5 mg/kg of body weight. In the above 91 cases, the negative conversion rate and the egg reduction rate of egg out-put before and after the drug administration were examined by the cellophane thick smear method and Stoll's egg counting technique. A follow-up examination was carried out 3-4 weeks after the drug administration. Other 59 cases of single and mixed infection of T. trichiurus and A. lumbricoides were examined by the cellophane thick smear method only and administered pyrantel tablets at a single dose of 2.5 mg/kg of body weight. The results were summarized as follows: The prevalence rates of T. trichiurus and A. lumbricoides infection in this study were 42.8% and 27.2% respectively by the cellophane thick smear method among 1,066 stool specimens. The egg negative conversion rate and the egg reduction rate in the administration of oxantel/pyrantel tablets at a single dose of 20 mg/kg body weight were 75.0% and 97.8% respectively in T. trichiurus infection and 100% each in A. lumbricoides. The egg negative conversion rate and the egg reduction rate in the administration of oxantel suspension at a single dose of 15 mg/kg body weight were 85.7% and 93.0% respectively in sinlge infection of T. trichiurus. The egg negative conversion rate and the egg reduction rate in the administration of pyrantel dry syrup at a single dose of 5 mg/kg body weight were 100% each in single infection of A. lumbricoides. The qualitative evaluation in cases of single and mixed infection of T. trichiurus and A. lumbricoides were by means of pyrantel tablets at a single dose of 2.5 mg/kg body weight, the cure being 100% against A. lumbricoides but 17.0% against T. trichiurus. There were no undesirable side effects in this study and oxantel suspension and pyrantel dry syrup were well accepted and tolerated.

A histopathologic study of the lungs infected with Paragonimus westermani in the dog

Pulmonary paragonimiasis is one of the most important endemic parasitic disease in Korea. Although it is well known that Paragonimus westermani invades into the lung tissue eventually resulting respiratory failure, the pathogenesis of inducing lung lesions is not fully elucidated. It is still debatable on the nature of morphological component of 'cyst' which is known to be the most consistent change in lung paragonimiasis. For the purpose of clarification of morphological changes associated with pulmonary paragonimiasis, an experimental study was carried out by giving metacercariae of P. westermani to 10 dogs. These dogs were sacrificed 2 months and 6 months after infection respectively, and light microscopic and electron microscopic observations were made. Following conclusions were obtained. Paragonimus lung lesions could basically be classified into two categories, i.e., direct mechanical effects by the worms and eggs, and the changes secondary to the worm infection. The relative importance of these two appeared almost same. Adult worms of P. westermani reside inside the lumina of the dilated bronchi in great majority of the cases. Only exceptions were secondary abscess formation and subsequent scar where no epithelial lining was detected around the worms. In 6 months group almost all P. westermani worms were found inside the bronchial lumina. Obliterative endobronchiolitis was another prominent feature among changes that were not directly associated with worms or eggs. This finding was more prominent in the distal segments of the bronchial trees that harbor the Paragonimus worms. P. westermani eggs were found as isolated and aggregated fashions, provoking characteristic granulomatous inflammation. Some of the egg granulomas were seen in the areas apart from the locations where the worms were physically located. Ultrastructural observations of the cyst wall showed fair numbers of plasma cells and mast cells. Some of the cystlining epithelial cells showed increased deposit of glycogen granules, suggestive of early metabolic alteration of the respiratory epithelium.

Immunoelectrophoretic studies of several helminths

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.

Lung findings in experimental paragonimiasis

A pathological study was done to elucidate sequential changes of the lungs in various time intervals following experimental paragonimiasis in 15 dogs and 15 cats. The dogs and cats were fed with 30-50 metacercariae of Paragonimus westermani, and were sacrificed at 15, 20, 30, 45, 60, 90 and 120 days after infection respecively. Autopsies were performed immediately after death. Gross and microscopic examination of the lungs showed following findings: There were no qualitative difference in pathological findings between dogs and cats. Pathological findings were first noticed at 20 days of infection in thoracic cavity, which consisted of fibrinous plueritis along with superficial hemorrhage. Although no worm was found in the lung parenchyma at this period, juveniles were seen in pleural cavity together with turbid effusion. Paragonimus juveniles were first recognized inside the lung parenchyma by 30 days of infection. This was the period when the lungs showed multiple areas of hemorrhage and probably active penetration by smaller worms. Hemorrhagic bronchopneumonia was quite pronounced from this stage through 45 days of infection. Paragonimus worm cyst was essentially composed of fibrous scar and heavy inflammatory cellular infiltrate. The lining epithelial cells were first became noticed by 2 months of infection. And these epithelial cells were thought to be probably transformed alveolar lining cells rather than bronchiolar epithelial cells. As the infection progress, the cyst wall became more stabilized and often showed squamous metaplasia. Fibrinous pleuritis with pleural effusion was very prominent finding in early periods of infection. Bronchiolitis and focal vascular sclerosis were often seen in experimental paragonimiasis.

Rhabditis elongata Schneider, 1866 from students in Korea

As the result of a fecal examination performed in October 1978 on 320 middle and high school students of Hapcheon, Korea, we found 2 cases of Rhabditis nematodes in 16-year old high school girls. Rhabditis nematodes isolated from their feces were cultured at 30 C by a filter paper culture method. After careful observation of male and female adults, we confirmed them as Rhabditis elongata Schneider, 1866 on the morphological basis of the male genitalia, and the papillae and measurements of various parts of the worm bodies.

A case of human thelaziasis in Seoul

Thelazia callipaeda, parasitic in the eye of 24 years old Korean male, was extracted from the left eye of the patient in Seoul, Korea. We confirmed that as T. callipeada morphologically, based on the descriptions of Hsu and other literature. We presents the 7th human thelaziasis case in Korea with the historical review of the literature.

Precipitin reactions between host organ extracts and antisera for Paragonimus and Clonorchis

Precipitin reaction between host organ extracts and antisera for Paragonimus and Clonorchis were studied to know whether the antigens from two species of trematodes have common characters with those of host organs. The brain, lung, heart, liver and muscle of rat and rabbit were extracted in 0.1 percent saline solution by freezing-thawing technique. For immunization of rabbit, the extracts of Paragonimus and Clonorchis was emulsified with equal amount of Freund's adjuvant and 1.0 ml of the mixture was injected subcutaneously once every 10 days for 5 times, and 3 days after the last immunization, antisera were prepared. Precipitin bands by agar-gel double diffusion methods were examined between organ extracts of rabbit or rat and Paragonimus or Clonorchis antisera. The results could be summarized as follows: Precipitin bands were appeared between extracts of liver, heart and muscle of rat and anti-Clonorchis rabbit sera but not appeared against brain and lung. Precipitin bands were appeared between extracts of lung, muscle and liver of rat and anti-Paragonimus rabbit sera but not appeared against brain and heart. Precipitin bands were appeared between extracts of liver and muscle of rabbit and anti-Clonorchis rabbit sera but not appeared against brain, heart and lung. Precipitin bands were appeared between extracts of lung and muscle of rabbit and anti-Paragonimus rabbit sera but not appeared against liver. The extracts of Paragonimus westermani showed precipitin bands against liver and brain of rat and liver of rabbit in agar-gel double diffusion.

Studies on the parasites of Korean amphibia. Trematodes of amphibians

An investigation on the trematodes of Korean amphibians was carried out during the period from April to September of 1976. A total of 883 amphibians of 6 species were examined. These frogs, toads and salamanders were collected in the areas of the Han river side. And also these amphibians comprised 775 Rana nigromaculata, 53 Rana rugosa, 21 Hyla arborea japonica, 2 Bufo bufo gargarizans, 2 Gallula tornieri and 30 Hynobius leechi. Out of the results of this work, 7 species of trematodes belonging to 2 orders were found as follows: Monogenea: Diplorchis ranae Ozaki, 1931. Digenea: Diplodiscus japonicus (Yamaguti , 1936), Gorgodera japonica Yamaguti, 1936, Haematoloechus sibiricus japonicus(Yamaguti, 1936) , Mesocoelium brevicaecum Ochi, 1929, Loxogenes liberum Seno,1980, Pleurogenoides japonicus (Yamaguti, 1936).

Indirect fluorescent antibody test for diagnosis of paragonimiasis

Indirect fluorescent antibody tests were performed with sera of paragonimiasis patients and skin test positive sera against Paragonimus antigen. Paragonimus antigen was prepared from lyophilized adult worms of P. westermani by defatting with ethyl-ether before extracting with barbital buffered saline. Preparation of Paragonimus antigen for the indirect fluorescent antibody test was based upon Sato's method used for sero-diagnosis of anisakiasis, with Sephadex G-25 instead of Sepharose 4B. The results were as follows: The indirect fluorescent antibody titers of paragonimiasis patient's sera ranged from 1:64 to 1:512, whereas the control sera showed titers of less than 1:16. As controls, Clonorchis patient's sera and parasite-free healthy human sera were used. In indirect fluorescent antibody tests, the skin test positive human sera against Paragonimus antigen showed a positive rate of 41.5 per cent in the case of titers more than 1:40. On the other hand, complement fixation tests on the same sera showed a positive rate of 32.5 per cent in the case of titers more than 1:20.