• Parasites, Hosts and Diseases
• /
• v.38 no.1
• /
• pp.17-23
• /
• 2000

Eosinophils are important effector cells in host defense against parasites. Excretory-secretory product (ESP) produced by helminthic worms plays important roles in the uptake of nutrients, migration in the host tissue, and in immune modulation. However little is known about the ability of the ESP to directly trigger eosinophil apoptosis. This study investigated whether the ESP of newly excysted metacercariae of Paragonimus westermani could induce apoptosis in human eosinophils. Apoptosis was assayed by staining the cells with FITC-annexin V, and the cells were analyzed by flow cytometry. It was found that the ESP of newly excysted metacercariae of P. westemani induced a direct time- and concentration-dependent increase in the rate of constitutive apoptosis in mature human eosinophils. Eosinophil apoptosis was first apparent 3 hr after treatment with the ESP and continued to increase after 6 hr of incubation with respect to the cells cultured in the absence of the ESP. While only 2.8% of the eosinophils incubated in the medium for 3 hr were apoptotic, 7.6%, 10.9% and 22.6% of the eosinophils treated with 10. 30 and $100{\;}\mu\textrm{g}/ml$ ESP were apoptotic, respectively. This result suggests that the ESP of newly excysted metacercariae of P. westermani directly induce eosinophil apoptosis, which may be important for the survival of the parasites and the reduction of eosinophilic inflammation in vivo.

• Parasites, Hosts and Diseases
• /
• v.38 no.3
• /
• pp.173-175
• /
• 2000

The present study examined postmetacercarial changes in the excysted metacercariae of Echinostoma caproni maintained in the defined medium Mixture 199 plus 20% calf serum for 7 days at $41^{\circ}C$. The gas phase was atmospheric air. Each culture was inoculated with 25 excysted metacerariae. Cultures were maintained upright in closed 15 ml plastic centrifuge tubes each containing 10 ml of medium plus 200 units of penicillin/ml and $200{\;}\mu\textrm{g}$ of streptomycin/ml. By 4 days in culture, most metacercariae had voided their excretory concretions. Organisms were clumped or solitary at the bottom of the cultures. Many organisms showed flaring of the oral collar and extension of both the collar and tegumentary spines. By 4 days in culture, posterior protuberances or bumps were noted on many of the organisms and some organisms showed abnormal vesicular growths or blebs at their posterior ends. Some mortality was noted in culture by day 5, but most organisms were still alive when the cultures were terminated on day 7.

• Journal of fish pathology
• /
• v.13 no.2
• /
• pp.137-142
• /
• 2000

Differences in larvicidal activity of complement and chemiluminescent response of phagocytes in carp (Cyprinus carpio), crucian carp (Carassius auratus) and false dace (Pseudorasbora parva) against excysted metacercariae of Clonorchis sinensis were investigated. The prevalence of C. sinensis metacercariae of false daces caught at Nakdong river (NR) was 100%. However, no C. sinensis metacercariae were found in false daces, which were collected at a reservoir in Chinyoung (CY), crucian carps and carps. The sera of false dace, which were intensively infected with C. sinensis metacercariae, killed excysted metacercariae of C. sinensis more readily than that of carp. However, the serum obtained from C. sinensis metacercariae-free false dace showed the lowest larvicidal ability. The larvicidal ability of sera collected from each fish species completely disappeared when the complement was inactivated by heating. When supernatant of excysted metacercariae homogenate were added to phagocytes of each species, the chemiluminescent responses were significantly (p<0.05) diminished in false dace and carp. The inhibition ratio of chemiluminescent responses by the supernatant was 22.9% in false dace, 9.6% in crucian carp and 12.4% in carp.

• Parasites, Hosts and Diseases
• /
• v.29 no.1
• /
• pp.43-54
• /
• 1991

An in vitro immune effector mechanism against the target encysted metacercariae of Paragonimus westermani was demonstrated in the rat system. Peritoneal exudate cells, mainly macrophages from normal rats, showed adherence to and killing of encysted metacercariae of p. westermani in the presence of complement-independent serum from rats infected with Paragonimus metacercariae. These reactions were specific for the excysted metacercariae, as tissue-migrating juvenile worms were not affected. Damage of encysted metacercariae of p. westermani due to antibody and macrophages was assessed by morphological observation, by cell adherence reaction and by the use of vital dyes. frypan blue dye exclusion proved to be a reliable indicator of judging metacercarial viability. Electron microscopic studies demonstrated that macrophages reacted with fusty material on the tegumental surface and fine structures in the syncytium of the parasites. The tubular tunnels formed between the basement membrane and muscle layers of the damaged parasites were also noticeable. The relevance of these findings to cellular immunity in the early paragonimiasis was discussed.

• Parasites, Hosts and Diseases
• /
• v.38 no.2
• /
• pp.103-106
• /
• 2000

It is well known that the cysteine proteases in excretory-secretory product (ESP) of Paragonimus westermani newly excysted metacercariae (PwNEM) are capable of degrading IgG in vitro. Recent evidence suggests that the IgG-coated surface, such as found on parasites, is one of the most effective physiologic stimuli for granulocyte activation. Therefore, this study was designed to investigate the effect of excretory-secretory product (ESP) of PwNEM on superoxide production of granulocytes stimulated with IgG The 96-well plates were coated with human IgG (0, 10, 30, $100{\;}\mu\textrm{g}/ml$) in the absence or presence of ESP. When granulocytes were incubated in the wells coated with human IgG in the presence of ESP, the level of superoxide production of granulocytes was reduced to about 90% when compared to the cells incubated in the wells coated with IgG alone. This inhibitory effect of the ESP on IgG-induced superoxide production of granulocytes was concentration-dependent. These results suggest that ESP secreted by PwNEM may be important in the control of effector functions of granulocytes stimulated with IgG in human paragonimiasis.

• Parasites, Hosts and Diseases
• /
• v.45 no.3
• /
• pp.199-204
• /
• 2007

Metacercariae of Gynaecotyla squatarolae (Digenea: Microphallidae) were discovered from the shore crab, Macrophthalmus dilatatus, purchased at a market in a coastal town of Taean-Eup, Chungcheongnam-do, Republic of Korea. Their adult flukes were confirmed by experimental infection of rats. The metacercariae of G. squatarolae were elliptical ($391.1{\times}362.5{\mu}m$), and the excysted metacercariae had progenetic genital organs, including the ovary and testes. To obtain adult flukes, 6 Sprague-Dawley rats were fed 500 metacercariae each, and killed at days 2, 4, and 6 post-infection. The adult flukes were identified as G. squatarolae (Yamaguti, 1934) Yamaguti, 1939, based on morphological characters, including 2 ventral suckers (1 large and 1 small), a large genital atrium equipped with the cirrus and the metraterm, separated male and female genital pores, a transversely long cirrus pouch, and extensive vitelline follicles. In the present study, it has been first proven that the shore crab M. dilatatus is a second intermediate host for G. squatarolae in the Republic of Korea.

• Parasites, Hosts and Diseases
• /
• v.36 no.4
• /
• pp.217-225
• /
• 1998

A scanning electron microscopic study was performed to observe surface ultrastructures of excysted metacercariae and adults of Metagonimus miyatai. Metacercariae were collected from the scale of the pale chub (Zacco platypus). and adult flukes were harvested 1-4 weeks after infection to rats. In excysted metacercariae, the oral sucker was devoid of tegumental spines and had type I and type II sensory papillae. Anteriorly to the ventral sucker, spines were dense and digitated into 5-7 points, whereas near the posterior end of the body spines were sparse and digitated into 2-3 points. In one-week adults, 7 type II sensory papillae were arranged around the lip of the oral sucker. and at inner side of the lip one pair of small and two pairs of large type I sensory papillae were seen on each side. The distribution of tegumental spines was similar to that of metacercariae, but they were more differentiated with 9-11 pointed tips. In two- to four- week old adults, the surface ultrastructure was nearly the same as in one-week old adults, however, sperms were frequently seen entering into the Laurer＇s canal. Conclusively, the surface ultrastructure of M. miyatai was generally similar to that of M. yokogawai, however, differentiation of tegumental spines and distribution of sensory papillae around the oral sucker were different between the two species. which may be of taxonomic significance.

• Parasites, Hosts and Diseases
• /
• v.34 no.4
• /
• pp.283-285
• /
• 1996

Six metacercariae were found from the gill filaments of Conger myriaster purchased at Mokpo-shi in korean on September, 1996. Based on the morphology of the excysted specimen. we identified them as metacercariae of Heterophyosis continua, C. myriaster is a new intermediate host H. continua in the literature.

• Journal of Veterinary Clinics
• /
• v.25 no.4
• /
• pp.307-309
• /
• 2008

The metacercariae species have often been isolated from fresh water fish in Korea. In this study, we reported Clinostomum complanatum, a species of metacercariae isolated from gill and muscle of Korean barbel (Hemibarbus mylodon), a natural monument fresh water fish in Korea, by morphological observation. The excysted metacercariae, tongue-shaped and progenetic, were 3.11-4.62 mm in length and 0.90-1.43 mm in width. Fresh water fishes are known as the second intermediate hosts of C. complanatum. This is the first isolation report from Korean barbel.

• Parasites, Hosts and Diseases
• /
• v.46 no.2
• /
• pp.95-99
• /
• 2008

Eosinophil degranulation plays a crucial role in tissue inflammatory reactions associated with helminth parasitic infections 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.