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.
Animals ; Antigens, Helminth/*analysis/isolation & purification ; Chromatography, Ion Exchange ; Immunohistochemistry/methods ; Microscopy, Immunoelectron ; Paragonimus/*immunology ; Support, Non-U.S. Gov't

Paragonimiasis is a food-borne parasitic zoonosis caused by infection with lung flukes of the genus Paragonimus. In Vietnam, research on Paragonimus and paragonimiasis has been conducted in northern and central regions of the country. Using a combination of morphological and molecular methods, 7 Paragonimus species, namely P. heterotremus, P. westermani, P. skrjabini, P. vietnamensis, P. proliferus, P. bangkokenis and P. harinasutai, have been identified in Vietnam. Of these, the first 3, P. heterotremus, P. westermani and P. skrjabini, are known to infect humans in other countries. However, in Vietnam, only P. heterotremus, found in some northern provinces, has been shown to infect humans. Even nowadays, local people in some northern provinces, such as Lai Chau and Yen Bai, are still suffering from P. heterotremus infection. In some provinces of central Vietnam, the prevalence and infection intensity of P. westermani metacercariae in freshwater crabs (the second intermediate hosts) are extremely high, but human cases have not been reported. Likewise, although P. skrjabini was found in Thanh Hoa Province, its pathogenicity to humans in Vietnam still remains uncertain. The results of molecular phylogenetic analyses of Vietnamese Paragonimus species provides new insights on the phylogeny and taxonomy of the genus Paragonimus. Comprehensive molecular epidemiological and geobiological studies on the genus in Vietnam and adjacent countries are needed to clarify the biodiversity and public health significance of the lung flukes.
Animals ; Humans ; Paragonimiasis/*epidemiology ; Paragonimus/classification/genetics/*isolation & purification ; Phylogeny ; Prevalence ; Sequence Analysis, DNA ; Shellfish/parasitology ; Vietnam/epidemiology

The present study investigated the infection status of Paragonimus westermani metacercariae in freshwater crabs (n = 363) and crayfish (n = 31) from October 2007 to October 2008 using the crush method. All of the freshwater crabs, Eriocheir japonicus, were negative for P. westermani metacercariae while 10 (32.3%) of the 31 examined crayfish were positive. The 10 positive crayfish were caught in Haenam, Jeollanam-do, and there were 8-59 (mean 28.4) metacrcariae per infected crayfish. These results suggest that P. westermani metacerariae are still transmitted by crayfish enzootically in southern Korea, and that freshwater crabs may transmit metacercariae only on rare occasions.
Animals ; Astacoidea/*parasitology ; Brachyura/*parasitology ; Humans ; Korea/epidemiology ; Paragonimiasis/epidemiology/parasitology/*veterinary ; Paragonimus westermani/*isolation & purification ; Prevalence

Paragonimiasis typically results from the consumption of raw or improperly cooked crustacea, especially crabs and crayfish. Although previously endemic in Korea, the prevalence of this disease decreased in the early 1970s because of educational campaigns and fewer intermediate hosts as a result of ecological changes. Recently, we were presented with a family where all members were infected with Paragonimus after ingestion of Kejang (= drunken crab). The mother was hospitalized for general myalgia and weakness first, followed by the father, who was hospitalized for dyspnea 2 month later. After the parents were diagnosed with paragonimiasis, we recommended their daughter to visit our hospital for a checkup, because they all had eaten freshwater crabs soaked in soybean sauce. She complained of generalized myalgia, fever, and pleuritic pain, and was also diagnosed with paragonimiasis. Peripheral blood of the 3 patients revealed hypereosinophilia, and computed tomography (CT) scans of their chests showed pleural effusion. The results of antibody tests by ELISA were positive for paragonimiasis. We report here the case series of familial paragonimiasis in a modern urban city, rather than in a typical endemic area.
Adult ; Animals ; Family ; Female ; Humans ; Korea ; Lung Diseases, Parasitic/*diagnosis/radiography ; Male ; Middle Aged ; Paragonimiasis/*diagnosis/radiography ; Paragonimus/isolation & purification

Environmental studies regarding Paragonimiasis have been done in Sulak-Myun, Kapyung-Goon (I district) and Yongmoon-Myun, Yangpyung-Goon (II district), both of which are considered "active" endemic areas, and in Byukje-Myun, Koyang-Goon (III district) an "arrested" area. 1. Paragonimiasis is still active in the Uhmos River, Sulak-Myun, Kapyung-Goon, where 49% of crayfish contained Paragonimus metacecaria. Snails which are the first intermediate host of Paragonimus westermani were moderately present. In the Oonke River, Yongmoon-Myun, Yangpyung-Goon, crayfish which are the second intermediate host of Paragonimus westermani, were found in every area of the river, but snails were not found in any area of either branch of the river. They were caught in areas which are located below the junction. In the Daeja river, Bykje-Myun, Koyang-Goon, crayfish were found only in the upper stream of the river and snails were not caught in any area of the river. 2. The results described above are attributed to the fact that snails, which have minimal mobility and vitality, were complete1y obliterated by the long, unprecedented drought in 1965. In contrust, crayfish with great mobility and vitality survived. Thus without the snail, the life cycle of Paragonimus western1ani become incomplete, and the clinical disease disappeared. 3. Rainfall is very important in maintaining water flow in the river, a necessary factor for the life cycle of Paragonimus westermani. Therefore we have charted tile amount of rainfall in the Seoul and Choonchun areas in 1965, compared with the average of the past 30 years, in order to ascertain the degree of drought which influnced the Paragonimus westermani hosts and cohabitants. 4. Geographic conditions which are also very important in maintaining water flow in small rivers was investigated. The conclusion was drawn, that a long river has an advantage over a short during drought; a tortuous river provides a better opportunity for miracidia of Paragonimus westermani eggs to migrate to snails, and for Pargonimus cercaria to invade crayfish than a straight rapidly flowing river. 5. We must conclude that man may ingest Paragonimus metacercaria by drinking water from shallow wells and streams in areas endemic to paragonimiasis, when we note that one crayfish of four caught in shallow wells in Kapyung-Goon contained many Paragonimus metacercaria. 6. Other factors are the increase in traffic due to the expanding population in Korean farm areas, and the increase in the use of farm chemicals, which seep into the rivers and reduce the number of snails, crayfish and crabs available as potential hosts. 7. In recent years, the wild fauna in the mountains, one of the important terminal hosts of Paragonimus westermani, have been markedly reduced in number. This may have contributed to decreasing paragonimiasis as directly infective to man. 8. Health education on paragonimiasis, informing the people of the dangers of eating raw or inadequately cooked crayfish and crabs, or those soaked less than 10 days in soysauce, and drinking the water from shallow wells and streams, is demonstrated to have decreased the incidence of this disease. 9. Because of the preceding facts, paragonimiasis in Kyunggi-Prvince is being decreased.
Animals ; Asian Continental Ancestry Group ; Crustacea ; *Disease Reservoirs ; *Disease Vectors ; Ecology ; Human ; Korea ; Paragonimiasis/*epidemiology ; Paragonimus/isolation & purification ; Snails

No abstract available.
Abdomen/pathology ; Animals ; Anthelmintics/therapeutic use ; Bronchi/pathology ; Humans ; Male ; Middle Aged ; Paragonimiasis/*diagnosis/drug therapy/parasitology ; Paragonimus westermani/isolation & purification ; Praziquantel/therapeutic use ; Tomography, X-Ray Computed

Paragonimiasis is a parasitic disease caused by the lung fluke, Paragonimus spp. Lung flukes may be found in various organs, such as the brain, peritoneum, subcutaneous tissues, and retroperitoneum, other than the lungs. Abdominal paragonimiasis raises a considerable diagnostic challenge to clinicians, because it is uncommon and may be confused with other abdominopelvic inflammatory diseases, particularly peritoneal tuberculosis, and peritoneal carcinomatosis. Also, subcutaneous paragonimiasis does not easily bring up clinical suspicion, due to its rarity. We herein report 2 cases of abdominal paragonimiasis and 1 case of subcutaneous paragonimiasis in Korea.
Abdominal Cavity/*parasitology ; Animals ; Biopsy, Fine-Needle ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Paragonimiasis/parasitology/*pathology/radiography ; Paragonimus/cytology/*isolation & purification ; Republic of Korea ; Subcutaneous Tissue/*parasitology

The cysteine proteases of Paragonimus westermani metacercariae are involved in metacercarial excystment, host immune modulation, and possibly in tissue penetration. In order to clarify the origin of the enzymes, 28 and 27 kDa cysteine proteases in metacercarial excretory-secretory products were purified through the FPLC system using Mono Q column chromatography. The polyclonal antibodies to the enzymes were produced in BALB/c mice. Immunolocalization studies revealed that both cysteine proteases were distributed at the linings of excretory bladder and excretory concretions of the metacercariae. It was suggested that the excretory epithelium of P. westermani undertake the secretory function of metacercarial cysteine proteases, in addition to its role as a route for eliminating waste products.
Animals ; Chromatography, Liquid ; Computational Biology ; Cysteine Endopeptidases/analysis/isolation & purification/*physiology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Paragonimus/anatomy & histology/*enzymology ; Support, Non-U.S. Gov't

Paragonimus harinasutai metacercariae were found in a species of freshwater crab, Indochinamon ou, collected in a small stream of Namback District, Luang Prabang Province, Lao PDR. Adult flukes were recovered after experimental infection of the metacercariae to dogs. Metacercariae were round or slightly elliptical, 0.666 x 0.626 mm in average size, and had a thin cyst wall of about 20 microm in thickness, a black excretory bladder, convoluted ceca, and some pinkish materials in the body. Adults were somewhat elongated, 95.2 x 36.5 mm in average size, covered with single-tipped tegumental spines, had a smaller oral sucker than the ventral sucker, a moderately branched ovary, and 5-6 lobulated testes. Eggs were ovoid and bilaterally symmetrical in shape, 79 x 45 microm in average size, and had a uniformly thickened shell. By the present study, it has been confirmed that I. ou is a new second intermediate host for P. harinasutai.
Animals ; Crustacea/*parasitology ; Disease Reservoirs/*parasitology ; Dogs ; Female ; *Host-Parasite Interactions ; Humans ; Laos ; Male ; Paragonimiasis/*parasitology ; Paragonimus/anatomy & histology/*isolation & purification/ultrastructure ; Shellfish/*parasitology

Eosinophil degranulation plays a crucial role in tissue inflammatory reactions associated with helminth parasitic nfections and allergic diseases. Paragonimus westermani, a lung fluke causing human paragonimiasis, secretes a large amount of cysteine proteases, which are involved in nutrient uptake, tissue invasion, and modulation of hos's immune responses. There is, however, limited information about the response of eosinophils to direct stimulation by cysteine proteases (CP) secreted by P. westermani. In the present study, we tested whether degranulation and superoxide production from human eosinophils can be induced by stimulation of the 2 CP (27 kDa and 28 kDa) purified from excretory-secretory products (ESP) of P. westermani newly excysted metacercariae (PwNEM). A large quantity of eosinophil-derived neurotoxin (EDN) was detected in the culture supernatant when human eosinophils isolated from the peripheral blood were incubated with the purified 27 kDa CP. Furthermore, the 27 kDa CP induced superoxide anion production by eosinophils in time- and dose-dependent manners. In contrast, the purified 28 kDa CP did not induce superoxide production and degranulation. These findings suggest that the 27 kDa CP secreted by PwNEM induces superoxide production and degranulation of human eosinophils, which may be involved in eosinophil-mediated tissue inflammatory responses during the larval migration in human paragonimiasis.
Animals ; Astacoidea/parasitology ; *Cell Degranulation ; Cysteine Endopeptidases/*immunology/isolation & purification ; Eosinophils/*immunology/metabolism ; Helminth Proteins/*immunology/isolation & purification ; Humans ; Paragonimiasis/*immunology/metabolism ; Paragonimus westermani/*enzymology/isolation & purification ; Superoxides/*immunology