• Journal of Life Science
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• v.17 no.4 s.84
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• pp.462-469
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• 2007

In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.

• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.187-196
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• 1989

The effects of gamma irradiation on the survival and development of C. sinensis metacercariae were studied to evaluate the feasibility of irradiation as a control measure for clonorchiasis. Pseudorasbora parve were collected at an endemic river of clonorchiasis and were used for irradiation of the fluke in three schemes. The first (Scheme 1) was irradiation of the isolated metacercariae from the fish followed by infection to experimental rats. The second (Scheme 2) was irradiation of the fish, and then the metacercariae were isolated and in(ected to rats. The third (Scheme 3) was irradiation on the rat livers after infection with normal metacercariae. Irradiation doses varied from 5 to 100 Gy for Schemes 1 and 2, and 10 to 25 Gy for Scheme 3. The rats were sacrificed 2 to 6 weeks after infection. In Scheme 1, the metacercariae irradiated at 50 Gy failed to survive in the rats after 2 or 6 weeks. However, 1 to 44% of the metacercariae irradiated at 5∼30 Gy survived. The estimated LD50 of Scheme 1 was 16.5 Gy. The flukes irradiated in Scheme 2 survived better than those in Scheme 1. The average worm recovery rate in 50 Gy was 28%(7∼39% individually). Increasing the dose up to 100 Gy brought a remarkably low survival rate of an average 1%(0∼3% individually). The LD50 of Scheme 2 was 47.5 Gy. Worm recovery rates in the 10 Gy group of Scheme 3 were 21∼39%, and those in the 25 Gy group were 2% and 34%. Although the metacercariae were irradiated, all of the recovered worms were morphologically normal. Only the worms irradiated with 10 Gy or 25 Gy after 9 days from infection in Scheme 3 showed underdeveloped testes and seminal receptacle. The present results suggest that irradiation of the fish by 100 Gy could be adopted as a control measure for clonorchiasis.

• Journal of Life Science
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• v.27 no.7
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• pp.739-745
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• 2017

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

• The Journal of Korean Medicine
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• v.20 no.1 s.37
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• pp.185-196
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• 1999

The aim of the present study was to investigate the effect of Hwangryeonheadock-Tang and Onchung-Eum on essential hypertension and hyperlipidemia. Rats were orally administered for 30days with Hwangryeonheadock-Tang and Onchung-Eum and the blood was withdrawn at 10, 20 and 30days after an oral administration. The heart rate, tail blood pressure, plasma renin activity, plasma level of aldosterone. catecholamine, sodium and angiotensin II were measured after an oral administration of Hwangryeonheadock-Tang and Onchung-Eum in spontaneously hypertensive rat(SHR). In addition, serum levels of total cholesterol, triglyceride, HDL-cholesterol. LDL-cholesterol and total lipid were measured cholesterol-fed rats. The results were summarized as follows ; 1. A significant decrease of tail blood pressure was shown at 10, 20 and 30days after Hwangryeonheadock-Tang and Onchung-Eum treatment in SHRs. compared with saline. 2. Heart rate was significantly decreased at 30days in SHRs after Hwangryeonheadock-Tang treatment and at 20, 30days after Onchung-Eum treatment in SHRs. compared with the effects of saline group. 3, A significant decrease of plasma aldosterone level was elicited at 10, 20days after Hwangryeonheadock-Tang treatment in SHRs, compared with the effects of saline group, 4. Plasma renin activity was significantly decreased at 10days after Onchung-Eum treatment compared with the effects of saline group in SHRs. 5. Plasma norepinephrine level was significantly decreased at 20 and 30clays after Onchung-Eum treatment in SHRs, compared with the effects of saline group. 6. A significant decrease of plasma epinephrine level was induced at 30days after Hwangryeonheadock-Tang treatment and at 10, 20 and 30days after Onchung-Eum treatment, compared with the effects of saline group in SHRs. 7. Plasma sodium level was. significantly decreased at 20days after Hwangryeonheadock-Tang and Onchung-Eum treatment, compared with the effects of saline group in SHRs. 8. Plasma angiotensin II level was significantly decreased at 30days after Onchung-Eum treatment, compared with the effects of saline group in SHRs. 9. A significant decrease of body weight was observed at 20 and 30days after Hwangryeonheadock-Tang treatment and at 10, 20 and 30days after Onchung-Eum treatment. compared with the effects of saline group in hyperlipidemia rats. 10. Hwangryeonheadock-Tang and Onchung-Eum showed a significantly decreasing effect at 30days on serum total cholesterol level in hyperlipidemia rats, compared with the saline treatment. 11. Hwangryeonheadock-Tang and Onchung-Eum saw 20 and 30days respectively on serum triglyceride level in the saline treatment. 12. Hwangryeonheadock-Tang and Onchung-Eum decreased on serum HDL-cholesterol level significantly, compared with the saline treatment in hyperlipidemia rats. 13. A significant decrease of serum LDL-cholesterol was observed at 10 and 30days after Hwangryeonheadock-Tang treatment and at 30days after Onchung-Eum treatment, compared with the effects of saline group in hyperlipidemia rats. 14. Hwangryeonheadock-Tang had a significantly decreasing effect at 10, 20 and 30days on serum total lipid level, compared with the saline treatment in hyperlipidemia rats. 15. Hwangryeonheadock-Tang elicited a significantly decreasing effect on weight of kidneys, spleen and testes respectively and Onchung-Eum induced on weight of liver and spleen respectively in hyperlipidemia rats, compared with saline treatment. These Findings suggest a possible anti-hypertensive and anti-hyperlipidemic effect of Hwangryeonheadock-Tang and Onchung-Eum.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.431-438
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• 2000

To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.147-159
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• 1998

A survey of the internal parasites in 115 fishes of Ophicephalus argus was carried out in Kyungpook province during the period from January 1995 to November 1997. A species of trematode from the stomach and a species of nematode from the caeca of the fishes was discovered respectively. The trematode was identified as Azygia hwangtsinyi (Tsin, 1993) and the nematode as Pingus sinensis ($Hs\ddot{u}$, 1993). The fluke was pressed at thick of 0.1mm~0.2mm between slide glasses and fixed in 70% ethyl alcohol solution. The fluke was washed with tap water after fixation and stained with hematoxylin-carmin and mounted in balsam through routine methods. The nematode was fixed in 5% formalin solution and mounted with lactophnol or glycerine jelly. Morphology : Arygia hwantsinyi ; The fluke is elongate body with approximately parallel margins and rounded extremities in pressed preparations(Fig 1). The cuticle is unarmed. The oral sucker is ventro-subterminal in position. The ventral sucker is slightly smaller than oral sucker and positioned at 14.2% of body length from the oral sucker. The cirrus sac and genital pore is in mid-ventral line, slightly anterior to the ventral sucker. The oral sucker is succeeded by a well developed muscular pharynx. It is succeeded by a short inverted Y-shaped oesophagus. The inner wall of oesophagus is consisted of many fine folded membrane. The fine tubes of esophagus pass into the intestinal heads which are distended and presented a marked borderline between the fine tube and intestinal heads. The Intestinal heads start at level of pharynx and pass caudad in a zigzag course to end quite near caudal margin. The uterine coils occupy the intercecal area between the level of the cephalic margin of the ovary and about the level of the ventral sucker. The uterus is filled with many eggs. The testes are round or oval and placed one behind the other generally a little diagonally. The posterior testis is placed at level of 75.1% of body length. The ovary is oval, placed ahead the anterior testis. The shell gland not discernible outline is contacted with ovary ahead it. The vitelline glands are consisted of small oval vesicles. The vesicles occupy extracecal regions and between a little posterior of the ventral sucker and near caudal margin. Pingus sinensis : Female ; The cuticle is smooth and the cervical alae are well developed. The mouth opening is small, triangular, and without lips. The anterior end bears four sub-medial papillae. The oesophagus is divided into a muscular and glandular portion. The nerve ring is situated just at the junction of the muscular and glandular portions of the oesophagus. The vulva which has a prominent flap, is placed behind the middle of the body about one-third of the length of the worm from the posterior extremity. The vagina is strongly muscularised and after running anteriorly about $68{\mu}$, divides itself into two broad uterine tubes with an anterior and a posterior one. There are a few eggs in the uterine tubes. The tail is slender, straight and pointed. The ovary extends almost to the posterior extremity. Male : Male is smaller than female. The tail is pointed, curved and carrying well-developed caudal alae. A prominent muscular precloacal sucker is characteristic. The spicules are equal and short. There are four pairs of pedunculated pre-anal papillae, of which the most anterior pair are situated just at the level anterior to the muscular sucker. Two pairs of pedunculated papillae are present before the anus. There are five pairs of post-anal papillae of which the most anterior pair and the second counted from the posterior end are ventral and more marked than the ones which are situated laterally.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.361-371
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• 2000

It has been recently reported that 2-bromopropane (2-BP) induces male reproductive toxicity in both human and experimental animals. However, delayed effects of 2-BP on male reproductive system have not been investigated in detail. The present study was conducted to investigate the testicular toxicity of 2-BP and to determine the recovery of normal spermatogenesis in Sprague-Dawley rats. Male rats aged 5 weeks were administered 1,000mg/kg 2-BP by gavage daily for 4 weeks and sacrificed sequentially at 1, 2, 3, 4 and 12 weeks after initiation of 2-BP treatment. Testicular toxicity was evaluated qualitatively by histopathological examinations and quantitatively by reproductive organ weights, spermatid head count, and repopulation index. In the 2-BP treated rats, the body weights was significantly suppressed and the weights of testes and epididymides were also decreased in a time-dependent manner. On histopathological examination, spermatogonia in stages I-VI and preleptotene and leptotene spermatocytes in stages VII-IX were strongly depleted at 1 week of dosing. Spermatogonia were depleted extensively in all spermatogenic stages at 2 weeks. Continuing with the evolution of spermatogenic cycle, zygotene spermatocytes, pachytene spermatocytes, and round spermatids were sequentially depleted at 2, 3, and 4 weeks of dosing due to the depletion of their precursor cells. Vacuolization of Sertoli cells and spermatid retention were also observed at all time points, suggesting that 2-BP induced Sertoli cell dysfunction. At 12 weeks, after 8 weeks recovery, most of the tubules appeared severely atrophic and were lined by Sertoli cells only. Leydig cell hyperplasia in the interstitial tissue was also found. In addition, dramatic reductions in the number of spermatid heads and repopulation index were observed, indicating that 2-BP-induced testicular injury is irreversible. These results indicate that 4 weeks repeated-dose of 1,000mg/kg 2-BP results in a progressive germ cell loss due to the depletion of spermatogonia followed by long-term testicular atrophy in SD rats.

• Korean Journal of Veterinary Research
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• v.45 no.3
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• pp.325-334
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• 2005

Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.

• Journal of Life Science
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• v.19 no.7
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• pp.983-993
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• 2009

The effects of Monascus-fermented Korean red ginseng (MFRG) on the contents of serum lipids and tissues lipid peroxidation was investigated in alcohol feeding rats (AC group). Serum contents of total lipid and free fatty acid in alcohol feeding rats were significantly increased, but these increases tended to decrease in the AMFRG group. Serum triglyceride content was also significantly decreased in the AMFRG group compared to other groups. Serum content of total-cholesterol was significantly increased in AC group compared to normal control (NC) group, whereas there was no significant difference between the AC and AMFRG groups. Content of HDL-cholesterol in serum was slightly increased in the AC group compared to the NC group, but this increase in the AC group was more significantly increased in the AMFRG group. At the same time, atherogenic index (AI) was also significantly decreased in the AMFRG group compared to the AC group. Contents of thiobarbituric acid reactive substances (TBARS) in the liver, heart, spleen and testes were significantly increased in the AC group compared to the NC group, but these increases were significantly less in the AMFRG group. Contents of liver nonheme ion was increased in the AC group and was significantly decreased in the AMFRG group, which suggested that lipid peroxidation contents are inversely correlated with liver nonheme ion content. Hepatic glutathione concentration was significantly decreased in the AC group, but this content was significantly increased in the AMFRG group and it showed the antioxidant abilities of glutathione. These results suggested that Monascus-fermented Korea red ginseng has anti-atherogenic index (AI) effects as well as antioxidative activities through reduced tissue oxidative stress in alcohol feeding rats.

• Parasites, Hosts and Diseases
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• v.25 no.1
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• pp.69-82
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• 1987

The prevalences of the cuke belonging to genus Metagonimus hove been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Mepagonimus in the areas. The results obtained are summarized as follows: 1. The snails, Semisulcospira globus were collected (rom the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagenimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9%), Cercaria incerpa(0.6%), and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to dye oral spines on its anterior of the first line. 2. The cercariae of Metagonimus were experimentally exposed to goldfish. nfection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. 3. Seven species of ask were caught in the Daecheong Reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys widens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco termmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. 4. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3~65.8%) than those of the rats (3.5~31.6%). The flukes were found mostly in the middle and the lower part of small intestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. 5. The sixte of adult flukes varied by the hosts. In the adult cukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg sixte was $29.1{\times}1.7{\mu\textrm{m}}$. According to the above results, the cukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed.