Cultivation of the sparganum of Spirometra mansonoides in vitro with prolonged production of sparganum growth factor.
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Amnion↗
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Sparganum proliferum, a sparganum infected with a virus?
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Cestoda↗
[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↗
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↗
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↗
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↗
Sparganum proliferum: an overview of its structure and ultrastructure.
A detailed study of the structure and ultrastructure of Sparganum proliferum was made possible for the first time thanks to the successful in vitro and in vivo maintenance of this rare parasite. Although S. proliferum exhibits many of the classical tegumental and parenchymal structures previously described for other larval cestodes, these are either arranged in a distinct fashion or, in some cases, may be completely different. Among the latter and of special interest are the single or multiple parenchymal cavities, surrounded by tegument, which in some instances appear to act as a primitive digestive tract.
Animals↗
Maintenance of Sparganum proliferum in vitro and in experimental animals.
Successful in vitro and in vivo maintenance of Sparganum proliferum is described for the first time. Various experimental animals including hamsters, mice and a monkey were evaluated. Albino mice inoculated either subcutaneously or intraperitoneally allowed the survival and multiplication of larvae for as long as 72 weeks. Intensity of infection was proportional to the length of exposure; however, the number of larvae collected from inoculated animals varied widely when infection lasted for 6 or more months. Inoculation of single larval segments appears as effective as that of complete larvae. Although Minimal Essential Medium allowed the survival of S. proliferum for as long as 14 weeks, growth was observed only during the first 4 weeks of culturing. Despite initial in vitro growth of larvae, neither differentiation into a more developed stage nor multiplication was obtained.
Animals↗
The fate of spargana inoculated into the cat brain and sequential changes of anti-sparganum IgG antibody levels in the cerebrospinal fluid.
To establish an animal model of intracranial sparganosis, the fate and behavior of the experimentally inoculated spargana were observed. A total of 102 scolices of spargana were injected into 22 cat brains, and the cats were sacrificed at 2 weeks, 1 month, 3 months and 6 months after the inoculation. Neurosparganosis was established in 77% of the cats. Of 43 recovered worms, 19 (44%) were located in the subdural or subarachnoid space, 16 (37%) in the brain parenchyme, and 2 (5%) in the lateral ventricle. One was detected at the diploic space of the skull and 5 were outside the cranial cavity. All but one were alive, and had grown tails. They were distributed in the brain parenchyme randomly. There was no place which they could not invade. No adult was found in the intestine. Cerebrospinal fluid (CSF) was collected before inoculation, 1 week, 2 weeks, 1 month, 3 months and 6 months after inoculation. The level of anti-sparganum IgG antibody in CSF measured by ELISA began to increase above the criteria of positivity 1 month after inoculation. Three months after inoculation, the values markedly increased. The present findings reveal that intracranial inoculation of spargana into the brains of cats would be a good animal model of experimental neurosparganosis.
Animals↗
[State of Spirometra "sparganum growth factor" research].
The plerocercoids of all Spirometra forms investigated up to now in this regard produce a "Sparganum Growth Factor" (SGF), i.e. one (or more) substance(s) with growth hormone-like effect on certain rodents. The SGF produced by the species S. mansonoides (SGF "mansonoides") is stronger effect than the SGF produced by all the other forms (SGF "non-mansonoides"). The physiological in its effects hitherto investigated mainly with SGT "mansonoides" in the USA are shortly mentioned. In order to get further insights the isolation, purification, and characterization of the SGF are necessary. The both kinds of SGF are also of taxonomic importance.
Animals↗
[Two European Spirometra forms (Cestoidea: Diphyllobothriidae) with different Sparganum growth factors].
European Spirometra forms corresponding to S. erinaceieuropaei in morphology, host specificity and periodical destrobilation were tested for a Sparganum Growth Factor (SGF "mansonoides") acting upon hypophysectomized male rats. A Polish form as well as Thai and Burmese forms did not show a SGF "mansonoides". The Polish form partially caused increased growth in normal laboratory mice (similar to a form from Thailand) and in Cricetulus griseus (SGF "non-mansonoides", restricted by host resistance). Procyon lotor can act both as paratenic and as definitive host for the Polish form. The Polish, Thai and Burmese forms are attributed to S. erinaceieuropaei. A Bulgarian form showed a typical SGF "mansonoides". 4 hyposectomized 100 g rats, each of which were implanted 5 spargana subcutaneously, showed weight gains up to 400 g within 7 to 8 weeks. Therefore, this Bulgarian form is treated provisionally as Spirometra sp.
Animals↗
Disseminated parasitosis in an immunosuppressed patient. Possibly a mutated sparganum.
The autopsy of a man who died of Hodgkin disease revealed that a peculiar metazoan parasite had proliferated and disseminated throughout his body. The parasite could not be identified; however, electron microscopical studies revealed that it had the structure of a flatworm. This, together with its shape and structure, convinced us that the parasite was an aberrant sparganum manifesting uncontrolled proliferation and dissemination.
Blood Vessels↗
Purification of antigenic protein of sparganum by immunoaffinity chromatography using a monoclonal antibody.
The quality improvement of antigen (crude saline extract) of Spirometra mansoni pleroceroid (sparganum) was investigated by protein purification. The crude extract was fractionated by gel filtration through Sephacryl S-300 Superfine. Its third fraction was purified by affinity chromatography using a monoclonal antibody as ligand. When observed by SDS-PAGE, the purified protein was composed of 2 bands of 36 kDa and 29 kDa which were found already as the most sensitive components in the crude extract by immunoblots with patients sera. The quality of the purified antigen was evaluated in comparison with the crude extract by enzyme-linked immunosorbent assay (ELISA) for the specific (IgG) antibody in sera of human sparganosis, other parasitic and neurologic diseases, and normal control. When the purified antigen was used, the sensitivity was not altered but remained high (96.4%) while the specificity was increased from 86.8% to 96.9%.
Animals↗