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[Localization and characteristics of lactate and malate dehydrogenase in the sparganum and adult worm of Spirometra erinacei].

This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of S. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r-spargana (from experimentally infected rats). By isoelectrofocusing, isoelectric points (PI) of 4 MDH isozyme from adult worm were 6.0, 6.5, 6.7 and 7.1, respectively. PI 6.0 was the major band. The active range of pH for MDH was about pH 6 approximately 8 and the optimum pH was pH 7. The effective temperature on the MDH was about 30 degrees C approximately 50 degrees C and the optimum temperature was about 40 degrees C in spargana and adult worm. In the stability against heat, when MDH was heated at 85 degrees C for 10 seconds, the activity was denatured perfectly. Maximum activity of MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

Animals↗

[The Infection Status Of Sparganum And Gnathostoma In Frogs Of Southern Part Of Korea]

A survey on the infection status of sparganum and Gnathostoma in frogs was carried out in 11 areas in Gyeongsangnam-do and Cheonlanam-do province, a southern part of Korea. 1. Among 626 frogs examined, 4 percent were infected with sparganum, but none with Gnathostoma. The infection rates of sparganum were variable according to the areas. Those from Kimhae, Sunchun, Jinhae, Sachun, Kosung, Keoje, and Hadong were 16 %, 9.1 %, 9.1 %, 4.1 %, 2 %, 2 %, and 1.5 % respectively, but frogs from Hapchun, KwangJu, Namwon and Sanchung were not found infected. 2. Rana nigromaculata was the only species infected with sparganum. Rana rugosa and Bombina orientalis were not infected. 3. Sparganum was mainly found in the legs and the trunk of frog. About 92.5 % of worms were detected in the femoral intermuscular connective tissue. 4. Number per infected frog was in range of one to five, and 2.8 % of frogs were infected by only one worm. 5. The heavier the frog weight, the higher the infection rate of sparganum. In frogs of 51-60 grams, 28.6 % were infected.

Journal Article↗

Epidemiological significance of sero-positive inhabitants against sparganum in Kangwon-do, Korea.

Sparganum is a plerocercoid of pseudophyllidean tapeworms, Diphyllobothrium or Spirometra spp. Human sparganosis is endemic mainly in East and Southeast Asian countries where the custom of eating raw snake or frog meat, or poulticing with snake's skin exists. From January 1995 to November 1999, an epidemiological survey was undertaken to evaluate the serum levels of anti-sparganum specific IgG antibodies in Whachon-gun residents, Korea. An enzyme- linked immunosorbent assay (ELISA) and immunoblot analysis of the sera from 316 subjects were used. In addition, a stool examination from 416 inhabitants and questionnaires regarding the consumption of raw meat were given. Out of 416 inhabitants examined coprologically, one was infected with Clonorchis sinensis and two were infected with Metagonimus spp. The sera from 36 inhabitants (11.4 %) showed a positive reaction to the sparganum antigen. Out of these 36 inhabitants, the sera from 25 people were examined 7, 19, and 50 months later. The sera were found to still show positive reactions without any remarkable changes of anti-sparganum specific antibody titers except for one. An analysis of the questionnaires suggested that a history of eating of raw snakes or frogs was important risk factor for clinical or covert sparganosis (odd ratio=15.6 and 3.1, respectively).

Adolescent↗

[Prevalence of sparganum of frogs (Rana nigromaculata) in Dae-jeon area, Chung-nam, Korea]

Prevalence of sparganum in the frog (Rana nigromaculata) was surveyed in Dae-jeon city and in suburbs of the city, Chungnam Do. 39 (3.9 %) out of 1,011 frogs were infected with the worm, and the highest prevalence rate of sparganum was found the frogs from Shintanjin area(11.3 %). Infection rates of sparganum were observed due to the body weight and sex of frogs. The highest infection rate of frogs, the body weight of 50 gms or more, showed 30.3 per cent(female: 41.7 %, male: none), on the other hand, no worm was found in young frogs(less than 10 gms). Distribution of sparganum in the frogs was recorded and 82.6 % of worms were distributed in the femoral intermuscular connective tissues of the hind legs. Among those infected frogs, number of worm burden was one to five worms, and 53.3 per cent of frogs were infected by only one worm.

Journal Article↗

Sparganum infections in normal adult population and epileptic patients in Korea: a seroepidemiologic observation.

A seroepidemiologic observation of anti-Spirometra erinacei plerocercoid (sparganum) antibody (IgG) in serum was made in normal adult and epileptic patients in Korea from February, 1987 to September, 1990. Sera were tested by enzyme-linked immunosorbent assay (ELISA) for anti-sparganum antibody together with anti-Taenia solium metacestode, and anti-Paragonimus westermani antibodies. Sera reacted positively to sparganum antigen only were considered. Positive rate for anti-sparganum antibody in 850 normal adults was 1.9% (standardized rate by provincial population was 1.7%). In 2,667 randomly selected patients of epilepsy at 28 local centers of the Changmi Club, positive rate was 2.5% (standardized rate: 2.3%). In both normal adult and patient groups, the higher antibody rates were observed in Kangwon and Chonnam Provinces. Positive rates were 10 times higher in male than in female in normal adults and 4.5 times in male epileptic patients. The rates were elevated especially with age over 30-year. Odd ratio of the antibody was 1.32 which indicated an ambiguous etiologic factor for epilepsy.

Adolescent↗

[Immunohistochemical observation on the antigens inducing IgG and IgM antibodies against sparganum].

Localization and characterization of the antigenic components of sparganum which induced IgG and IgM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGE and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunized by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyma of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyma of sparganum and in the connective tissue of host. By 5-20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and IgM antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of excretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

Animals↗

Immunohistochemical localization of 36 and 29 kDa proteins in sparganum.

Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as primary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody (MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

Animals↗

Component proteins and protease activities in excretory-secretory product of sparganum.

Spirometra mansoni plerocercoid (sparganum) was incubated in saline at 4 degrees C or 37 degrees C up to 100 hours. Protein contents in the excretory-secretory product (ESP) were rather constant (mean 7.7 mg of protein/gram of sparganum) in the preparations. Reducing SDS-PAGE of ESP showed similar protein subunit compositions with those in crude extract. Antigenic 36 and 31 kDa proteins were major bands in ESP. ESP exhibited specific activities of protease (2.9-5.3 units/mg) at pH 6.0 and pH 7.5. Presence of protease activity in ESP may be a supporting evidence that hitherto known cysteine protease of sparganum is possibly secreted.

Animals↗

[Infectivity of the sparganum treated by praziquantel, gamma-irradiation and mechanical cutting].

An experimental study was performed to observe the infectivity of sparganum (plerocercoid of Spirometra erinacei) treated by praziquantel, gamma-irradiation and mechanical cutting. The spargana were obtained from the naturally infected European grass snake, Rhabdophis tigrina, or from the experimentally infected mice. A total of 83 mice (ICR strain) were divided into 3 experimental groups by the source of the damage, fed each with 5 spargana, and sacrificed 1 month later for worm recovery. In the praziquantel group, the worms were incubated in the concentration of 10 micrograms/ml (control: Tyrode for 4 hours) for 0.5, 1, 2 and 4 hours at 36 degrees C, and fed to mice. The recovery rate from mice in praziquantel group was not different from that (80%) of control group and in the range of 76-100%. In the gamma-irradiation group, the worms were irradiated by 10-1000 Gy with Cs137. The average recovery rates of 69-100% were not different from that of control up to 100 Gy. The rate was 56% under 150 Gy, and 5% by 1000 Gy. In the mechanical cutting group, the worms were cut at 0.5, 1, 2 and 3 mm from the anterior end of the scolex. The average recovery rates in each group were 70-90% and that of control was 90%. The present finding suggests that the sparganum be highly resistant to praziquantel, gamma-irradiation and mechanical cutting. The vitality center of the sparganum must be at the anterior end of its scolex.

Animals↗

Comparison of carbohydrate moieties of sparganum proteins of the snake, mouse and those of adult worm.

The carbohydrate moieties of larval sparganum proteins in two different species, the snakes, Elaphe rufodorsata, the Balb/c mouse and those of the adult worm, Spirometra erinacei, were compared using five different lectins including GNA, SNA, MAA, PNA and DSA. The GNA positive 53 kDa molecule, which is excretory-secretory protease in the sparganum from the snake showed a stage specific and developmental regulation. We also suggested that sparganum glycosylation may be involved in immune evasion and differentiation into an adult worm.

Animals↗

The changes of histopathology and serum anti-sparganum IgG in experimental sparganosis of mice.

The present study is intended to observe the chronologic changes of experimental sparganosis by histopathological observation and detection of circulating anti-sparganum IgG antibody using ELISA. Each of 25 mice was infected with five spargana, and they were examined after 1, 2, 4, 10 weeks or 6 months from infection. The followings are summarized results. 1. The plerocercoids were detected in the subcutaneous tissue of the trunk, neck or axilla, but a few often extended into the skeletal muscle. The recovery rates were 72% at the first week, 80% at the second week, 95% at the fourth week, 92% at the tenth week and 100% at the sixth month. The larvae grew slowly in both length and weight until 6 months. 2. Histopathologically, most of the larvae were observed alive in the soft tissue or skeletal muscle. Numerous eosinophils, neutrophils, lymphocytes and plasma cells were infiltrated focally around the worms by the second week, but they surrounded the worms to form a layer of inflammatory reaction after 4 weeks of infection. Also histiocytes and fibroblasts began to appear around the inflammatory cells at 4 weeks. After 10 weeks, the worms encircled by a thin fibrous layer were found. After 6 months, the worms were surrounded by either fibrous tissue or active inflammatory cells. The inflammation looked more severe in the tracks left by the worms, rather than around the worms. 3. The level of anti-sparganum IgG antibody in the serum showed an increase by the fourth week, and a rapid and continuous increase was observed thereafter by the tenth week after infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography.

Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum), 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

Animals↗

Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation.

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

Animals↗

Modification of carbohydrate compositions of 31/36 kDa proteins of plerocercoids (sparganum) of Spirometra mansoni grown in different intermediate hosts.

We purified specific 31/36 kDa antigenic molecules from sparganum in different intermediate hosts (snakes and mice) and analyzed their monosaccharide compositions. Compositional analysis showed that glucose and mannose concentrations were 2-3 fold higher in the 31/36 kDa molecule purified from snakes than those from mice. This result implies that antigenic glycoproteins of sparganum from snakes might be modified in mammalian sparganosis with respect to their carbohydrate composition.

Animals↗

Granulomatous mastitis caused by sparganum. A case report.

BACKGROUND: Sparganosis is not a common disease, and its rarity makes it difficult to distinguish from others. Close examination of fine needle aspiration biopsy (FNAB) samples, however, can aid in histologic examination of the lesion. CASE: A 64-year-old female, born and raised in a rural area in the state of Goiás, Brazil, presented with a relatively mobile and hard lump in the lower inner quadrant of the right breast. It was initially diagnosed as granulomatous mastitis, but cytologic examination of the surgical specimen revealed a sparganum larva, also revealed by anterior FNAB. CONCLUSION: Sparganum should be considered in the differential diagnosis of granulomatous mastitis, especially among Asians living in other countries.

Biopsy, Needle↗

On The Sparganum Mansoni Infection In Some Korean Terrestrial Snakes.

Distribution of Sparganum mansoni in 7 species of terrestrial snakes in Wonju City was surveyed. All kinds of snakes were found to be served as intermediate hosts of this larval worm except Elaphe schrenkii which has already been recorded as important host. Authors believe that Zamenis spinalis was firstly recorded as intermediate host of Sparganum mansoni in Korea. Some aspects of infection status, host-parasite relations and sources of human infection were briefly discussed.

Journal Article↗